Clostridia

Med J Aust, 1980 Apr 5, 1(7), 321 - 3
Relapse of antibiotic-associated colitis after vancomycin therapy; Wilkinson IJ et al.; Antibiotic-associated colitis, although occasionally fatal, is a disease which is considered to be self-limiting and non-recurring . Recently, specific treatment with oral vancomycin directed at the trigger organism, Clostridium difficile, has been shown to be effective . A case in which antibiotic-associated colitis was treated with vancomycin and subsequently recurred is described . The fact that such relapse can occur indicates that further evaluation of the efficacy of vancomycin is required.

Jpn J Med Sci Biol, 1980 Apr, 33(2), 81 - 6
The susceptibility of the mallard duck (Anas platyrhynchos) to Clostridium botulinum C2 toxin; Jensen WI et al.; Most strains of Clostridium botulinum type C, after having lost their capacity to produce their dominant toxin (C1) as a result of being "cured" of their prophages, continue to produce C2, a trypsin-activable toxin reported by other investigators . While of relatively low toxicity when administered perorally to the adult mallard duck (Anas platyrhynchos), it was highly toxic when given parenterally . By the intravenous route, for example, it was more than 1,000 times as toxic as C1 toxin by the same route, when compared on the basis of mouse intraperitoneal toxicity . The cause of death in every instance was massive pulmonary edema and hemorrhage rather than the respiratory paralysis that occurs in C1 intoxication.

Am J Vet Res, 1980 Apr, 41(4), 650 - 3
Protective cellular antigen of Clostridium chauvoei; Stevenson JR et al.; Cellular antigens of Clostridium chauvoei, strain IRP-128, were demonstrated to be important in induction of immunity against this bacterium in guinea pigs . At least one major component of the cellular antigen complex was heat-labile . Acid extraction of the bacterial cells, followed by selective purification for flagella, led to the preparation of an acid extract antigen that possessed a high degree of immunogenicity . The acid extract antigen contained flagellar components and was resolved into two major and approximately five minor protein components by polyacrylamide-gel electrophoresis.

Antimicrob Agents Chemother, 1980 Apr, 17(4), 695 - 8
In vitro susceptibility of Clostridium difficile isolates from patients with antibiotic-associated diarrhea or colitis; Dzink J et al.; In vitro susceptibility tests were performed on 84 strains of Clostridium difficile to 11 antimicrobial agents . All isolates were from the stools of patients with antibiotic-associated diarrhea or colitis in which there was a cytopathic toxin that was neutralized by Clostridium sordellii antitoxin . Over 95% of the strains were susceptible to vancomycin, penicillin G, ampicillin, and metronidazole at concentrations of 4 microgram/ml . Susceptibility to clindamycin was variable; 60% of the strains were susceptible at 1 microgram/ml, and 9% were resistant at 128 microgram/ml . Studies of individual isolates showed that a major portion of the strains were relatively susceptible to the antimicrobial agent implicated in causing the disease.

Infect Immun, 1980 Apr, 28(1), 277 - 82
Clostridium difficile in gnotobiotic mice; Onderdonk AB et al.; Germfree mice associated with Clostridium difficile developed intestinal disease characterized by polymorphonuclear cell infiltration of the lamina propria, diarrhea, and cecal cytotoxin concentrations positive at a 10(-6) dilution . The numbers of viable bacteria never exceeded 10(10) colony-forming units per g (dry weight) . Despite the high toxin levels and chronic inflammation over a 30-day period, the mortality rate was low (less than 2%) . Daily treatment of these animals with two oral doses of 2 mg of vancomycin resulted in stool levels of greater than 200 micrograms/ml, well in excess of the minimum inhibitory concentration for C . difficile . This therapy decreased viable cell density by 2 to 3 logs and increased the spore counts from 10(5.8) to 10(7.8) colony-forming units per g (dry weight) by day 7, and animals were free of detectable toxin . However, once therapy was stopped, viable bacteria and spore counts and cytotoxin concentrations returned to previous levels . Treatment of mice with concentrations of clindamycin shown to be inhibitory in vitro had no effect on C . difficile toxin titers or bacterial counts, although the appearance of a clindamycin-resistant population was noted . These data indicate that vancomycin, given orally, decreases the concentration of toxin, but C . difficile survive as spores . By contrast, large populations of vegetative cells and high cytotoxin levels persist when clindamycin is used, even at an inhibitory concentration.

Can J Microbiol, 1980 Apr, 26(4), 448 - 53
Production of indole-3-propanoic acid and 3-(p-hydroxyphenyl)propanoic acid by Clostridium sporogenes: a convenient thin-layer chromatography detection system; Jellet JJ et al.; Indole-3-propanoic acid (IPA), 3-(p-hydroxyphenyl)propanoic acid (HPPA), and 3-phenylpropanoic acid (PPA) were present in the spent bacterial media of Clostridium sporogenes (20/20 strains) and C . cylindrosporum (1/1 strains), but absent in 32 other clostridial species (74 strains) tested . Both IPA and HPPA (but not PPA) could be readily detected by thin-layer chromatography and p-hydroxybenzaldehyde spray reagent . IPA forms a scarlet complex with p-hydroxybenzaldehyde which shifts to purple and remains stable for up to 6 weeks . IPA can be detected in acidified extracts of C . sporogenes by a simple spot test . The structures of IPA and HPPA were confirmed by nuclear magnetic resonance (nmr) and mass spectroscopy and their formation was detected by the absorbance at 280 nm . Addition of one of the precursor amino acids (L-tryptophan, L-tyrosine, or L-phenylalanine) to the medium greatly enhanced formation of the corresponding deaminated acid and depressed the formation of the other two acids . The products IPA, HPPA, and PPA, at 10(-3) M, and spent bacterial media were negative in the direct Ames's assay for mutagenicity and noncytotoxic towards MRC-S cells.

Arch Intern Med, 1980 Apr, 140(4), 574 - 6
Cephalosporin-associated colitis and Clostridium difficile; Donta ST et al.; A case of cephalosporin-associated colitis occurred in which a tissue-cultured morphologic-altering activity was demonstrated in the patient's feces during the active episode . Neutralization of the tissue culture activity by antiserum directed against a partially purified toxin of Clostridium difficile provided a more suggestive link between the colitis and this clostridial species.

Appl Environ Microbiol, 1980 Apr, 39(4), 764 - 9
Distribution of Clostridium botulinum; Huss HH; The distribution of Clostridium botulinum in the natural environments of Denmark, The Faroe Islands, Iceland, Greenland, and Bangladesh was examined . A total of 684 samples were tested . Type E was found in 90% of samples from the aquatic environment of Denmark, including sediments from young artificial lakes, and in 86% of samples from the marine environment of Greenland . Type E was not found in Danish cultivated soil and woodlands, including cultivated soil from reclaimed sea beds, but type B was frequently demonstrated in these environments . C . botulinum types A, B, or E were found in 2.6% of samples from the environments of the Faroe Islands and Iceland, whereas types C or D were demonstrated in 42% of samples from Bangladesh . The incidence of type E in aquatic sediments was not related to general industrial pollution or a high content of rotting vegetation . Fish or a rich aquatic fauna, on the other hand, appeared to contribute to a high incidence of type E . Based on these findings, it is suggested that type E is a true aquatic organism, because this environment offers the best conditions for survival of the spore in nature . It is further suggested that its presence in aquatic bottom deposits is based on sedimentation after proliferation in the carrion of the aquatic fauna and dissemination by water currents and migrating fish.

Lab Anim Sci, 1980 Apr, 30(2 Pt 1), 241 - 4
Acute gastric dilatation in monkeys: a microbiologic study of gastric contents, blood and feed; Bennett BT et al.; Twenty-one of 24 simian primates with acute gastric dilatation had Clostridium perfringens in their gastric contents . Only 2 of 18 normal animals contained this organism in their gastric contents . Clostridium perfringens was isolated from monkey biscuits taken from the cages of five affected animals and from five of 11 incoming lots of feed . After these biscuits were fed to normal animals, this organism could be isolated from the gastric contents . There were no other organisms isolated which could account for the voluminous gas production in this condition.

Methods Find Exp Clin Pharmacol, 1980 Apr, 2(3), 145 - 50
Comparison of metronidazole assay by microbiological and chemical methods; Bergan T et al.; Chemical (thin layer chromatography/fluorescence quenching in situ) and microbiological (agar well, diffusion technique with Clostridium perfringens as indicator strain) methods of assaying metronidazole have been compared . On dummy samples made with pure metronidazole in pooled human serum, both methods had a coefficient of variation ranging from 5.5 to 9.6 per cent of the mean . The microbiological method slightly underestimated the real amounts, and also had lower values than the chemical procedure . Comparison of serum and urine samples taken during the early, middle and late periods after medication to volunteers showed that biotransformation to antibacterially active metabolites contributes significantly to the antibacterial activity, particularly in urine . Biotransformation explains why microbiologically determined concentrations were higher than those determined chemically in samples taken at least 16-20 hours after intake of tablets or suppositories . It is important to be aware of the circumstance that the results of microbiological assay are sensu strictu limited to the particular indicator strain used, since other bacteria may exhibit other patterns of sensitivity to metronidazole and its metabolites.

Avian Dis, 1980 Apr-Jun, 24(2), 324 - 33
Role of Coccidia in the occurrence of necrotic enteritis of chickens; Al-Sheikhly F et al.; Clostridium perfringens type A, Eimeria acervulina, and Eimeria necatrix were used to produce necrotic enteritis in chickens . The disease was produced in all groups of birds that received feed contaminated with C . perfringens . Mortality due to necrotic enteritis was highest (53%) in birds infected with E . acervulina before infection with clostridia . There was a significant difference in mortality rates between birds infected with E . acervulina and birds infected with E . necatrix before infection with C . perfringens . Mortality rates also differed significantly between the group infected with E . necatrix and the group that received only feed contaminated with C . perfringens . It was concluded that under field conditions, coccidia can play a significant role in the occurrence of necrotic enteritis when a sufficient number of toxigenic strain of C . perfringens type A is present . The pathological changes induced by clostridia and coccidia are described.

Aust Vet J, 1980 Apr, 56(4), 181 - 3
The isolation of Salmonella from jejunal and caecal lymph nodes of slaughtered animals; Moo D et al.; One jejunal and one caecal lymph node were sampled from each of 50 cows, 40 yearling cattle, 25 sheep, 20 lambs and 45 pigs after slaughter . Salmonella, Clostridium perfringens and Staphylococcus aureus, all organisms which cause food poisoning in man, were sought by direct plating methods . The samples were also enriched and cultured for Salmonella . Organisms were cultured from 208 (58%) of the 360 lymph nodes; aerobic plate counts yielded up to 25,000 organisms per gram of tissue, although from most infected samples less than 1000 organisms per gram were cultured . Salmonella was isolated directly from 5% of samples, with counts up to 1,500 per gram . After enrichment Salmonella was isolated from nodes taken from 15 cows, 2 yearling cattle, one sheep and 8 pigs . Cl . perfringens was isolated from the caecal nodes of 2 yearling cattle and 2 pigs; S . aureus was not isolated from any sample . It was concluded that mesenteric lymph nodes may be a significant reservoir of Salmonella for transfer to meat and meat products.

Arch Fr Pediatr, 1980 Apr, 37(4), 233 - 40
{Clostridium perfringens infection and necrotising enterocolitis}; Loc C et al.; During a four year period (1974--1977) 21 infants died as a result of severe necrotising enterocolitis (N.E.C.) . In 9 cases, Clostridium perfringens was isolated . When this organism is recovered either from the placenta or from the first meconium and or when the signs of the disease appear within a few days of birth, materno-fetal transmission of the infection may be suspected . The infection occurs most frequently in neonates in whom the gastrointestinal tract was already colonized by Clostridium.

J Clin Pathol, 1980 Apr, 33(4), 395 - 9
Synergistic haemolysis test for presumptive identification and differentiation of Clostridium perfringens, C . bifermentans, C . sordellii, and C . paraperfringens; Gubash SM; A new test for the presumptive identification of Clostridium perfringens, C . bifermentans, C . sordellii, and C . paraperfringens is described . The test is based on the synergistic haemolysis shown by the clostridia and group B streptococci on sheep and human and CaCl2-supplemented human blood agar . C . perfringens gave crescent-shaped synergistic lytic zones (7 to over 20 mm in length), and C . paraperfringens usually small-sized (3 mm), bullet-shaped reactions on all three types of media . C . bifermentans showed a horseshoe-shaped synergistic reaction only on human blood containing media, and C . sordellii only on CaCl2-supplemented human blood agar . C . perfringens type A antiserum inhibited synergistic lytic activities of the four species . The test provided a reliable method for presumptive identification and differentiation of the four clostridial species and may obviate the need for the Nagler test.

Acta Pathol Microbiol Scand {B}, 1980 Apr, 88(2), 95 - 102
Binding of enterotoxin from Clostridium perfringens type A to liver cells in vivo and in vitro . The enterotoxin causes membrane leakage; Skjelkvale R et al.; Enterotoxin from Clostridium perfringens was shown to retain its biological activity after labelling with 125I . When injected intravenously into mice and rats, most of the radioactivity in the organs was present in the form of intact toxin . Studies of the tissue distribution of labelled enterotoxin showed the largest amounts in the liver, where the activity reached a maximum 10--15 min after administration . The highest concentration per g tissue was found in liver and kidneys . The radioactivity was excreted in the urine as a mixture of intact labelled toxin and low molecular weight degradation products . In vitro studies with purified parenchymal liver cells showed rapid release of lactate dehydrogenase (LDH) during treatment with enterotoxin, thus indicating severe membrane damage.

J Bacteriol, 1980 Apr, 142(1), 262 - 7
Phosphatidylglycerophosphate synthease and phosphatidylserine synthase activites in Clostridium perfringens; Carman GM et al.; Cytidine 5'-diphospho (CDP)-1,2-diacyl-sn-glycerol (CDPdiacylglycerol):sn-glycerol-3-phosphate phosphatidyltransferase (EC 2.7.8.5, phosphatidylglycero-P synthase) and CDPdiacylglycerol:L-serine O-phosphatidyltransferase (EC 2.7.8.8, phosphatidylserine synthase) activities were identified in the cell envelope fraction of the gram-positive anaerobe Clostridium perfringens . The association of phosphatidylglycero-P synthase and phosphatidylserine synthase with the cell envelope fraction of cell-free extracts was demonstrated by sucrose density gradient centrifugation, by both activities sedimenting with the 100,000 x g pellet and solubilization of both activities from the 100,000 x g pellet with Triton X-100 . The pH optimum for both enzyme activities was 8.0 with tris(hydroxy-methyl)aminomethane-hydrochloride buffer . Phosphatidylglycero-P synthase activity was dependent on magnesium ions (100 mM) . Phosphatidylserine synthase activity was dependent on manganese (0.1 mM) or magnesium ions (50 mM) . Both enzyme activities were dependent on the addition of the nonionic detergent Triton X-100 . Maximum phosphatidylglycero-P synthase and phosphatidylserine synthase activities were obtained when the molar ratio of Triton X-100 to CDP-diacylglycerol was 50:1 and 12:1, respectively . The Km for sn-glycero-3-P in the phosphatidylglycero-P synthase reaction was 0.1 mM . The Km for L-serine in the phosphatidylserine synthase reaction was 0.15 mM . Both enzyme activities were 100% stable for at least 20 min at 60 degrees C.

J Hyg (Lond), 1980 Apr, 84(2), 191 - 202
Commercial milk products and indigenous weaning foods in a rural West African Environment: a bacteriological perspective; Barrell RA et al.; Two commercially available baby milks, one 'biologically acidified', the other 'non-acidified', and a traditional weaning food, millet gruel, were prepared and stored under village conditions in West Africa . Increases in total colony count and in number of Bacillus cereus, Clostridium welchii, Staphylococcus aureus and Escherichia coli were determined in these products when stored as commonly practised at ambient temperatures over a period of 8 h . Poor hygiene during preparation was indicated by readily detectable numbers of coliforms and E . coli in freshly prepared samples of each of the milks, though the cooked local gruel seemed less vulnerable in this respect . The rate of increase in the numbers of these organisms was lower in the acidified milk when prepared with unboiled water containing high numbers of coliforms and E . coli . Increases in total colony count and in numbers of Staph . aureus were also less marked in the acidified milk . When food was not eaten soon after preparation the problem of bacterial overgrowth was as great with the local gruel as with the considerably more nutritious reconstituted milks.

Arch Microbiol, 1980 Apr, 125(3), 221 - 5
Lindane degradation by cell-free extracts of Clostridium rectum; Ohisa N et al.; For lindane degradation, a cell suspension of Clostridium rectum strain S-17 demands the addition of substrates such as leucine, alanine, pyruvate, a leucine-proline mixture, and molecular hydrogen . In the presence of leucine-proline mixture, lindane decomposed in parallel with isovaleric acid formation, and both lindane degradation and isovaleric acid formation were inhibited by monoiodoacetic acid, suggesting a close relation between lindane degradation and the Stickland reaction . Lindane was degraded by cell-free extracts of C . rectum in the presence of dithiothreitol (DTT) . Radiogaschromatograms of n-hexane soluble metabolites from {14C} lindane showed the presence of monochlorobenzene and gamma-3,4,5,6-tetrachlorocyclohexene . Leucine, NADH, and NADPH were somewhat less active than DTT for lindane degradation in cell-free extracts . Reductive dechlorination seemed the major route of lindane degradation in cell-free extracts as well as in the intact cells of C . rectum.

Biochim Biophys Acta, 1980 Mar 20, 628(3), 328 - 35
Interaction between Clostridium botulinum neurotoxin and gangliosides; Kitamura M et al.; The effect of gangliosides on Clostridium botulinum type A neurotoxin was examined in terms of detoxification . The molar concentrations of gangliosides necessary to detoxify 50% of 1 M Cl . botulinum neurotoxin were as follows: GM1, 2073; GM2, 2439; GM3, 6098; GD1a, 610; GD1b, 488; GT1a, 829; GT1b, 6 and GQ1b, 27 . Inhibition by gangliosides of the neurotoxin binding to synaptosomes showed that GT1b was highly effective, but the others were not . Low-temperature treatment inhibited the detoxification of neurotoxin by GT1b and the binding of 125I-labelled neurotoxin to the synaptosome fraction . 125I-labelled neurotoxin was mixed with GM1 or GT1b and their molecular size was estimated by sucrose-density-gradient centrifugation . When 125I-labelled neurotoxin was incubated with GM1, a single radioactive peak having a sedimentation coefficient of 7.3 S appeared . When incubated with GT1b, however, 125I-labelled neurotoxin gave three peaks having sedimentation coefficients 14, 10 and 7.3 S, respectively . The present results indicated that the location and the number of sialic acids in ganglioside molecules are of significance in the detoxification and the binding of Cl . botulinum neurotoxin with ganglioside molecules.

Carbohydr Res, 1980 Mar 15, 81(2), 315 - 22
Rapid procedures for determination of endo-N-acetyl-alpha-D-galactosaminidase in Clostridium perfringens, and of the substrate specificity of exo-beta-D-galactosidases; DiCioccio RA et al.; Culture fluid of Clostridium perfringens hydrolyzed the synthetic, chromogenic substrates beta-Gal-(1 leads to 3)-alpha-GalNAc-1 leads to OPh and beta-Gal-(1 leads to 3)-alpha-GalNAc-1 leads to OC6H4-NO2-o or -p to beta-Gal-(1 leads to 3)-GalNAc and the aglycon . Such assays facilitated the characterization and purification of this endo-N-acetyl-alpha-D-galactosaminidase activity . This activity was purified 1200-fold by fractionation with ammonium sulfate and chromatography on columns of Sephadex-G200, DEAE-Sephadex, and hydroxylapatite . The final preparation showed activity over a broad range of pH, with an optimum at 9.0, but less-pure material had two pH optima, 4.0 and 9.0 . Another assay method, which employed the synthetic, chromogenic substrates beta-Gal-(1 leads to 3)-beta-GlcNAc-1 leads to OC6H4NO2-p, beta-Gal-(1 leads to 4)-beta GlcNAc-1 leads to OC6H4NO2-p, and beta-Gal-(1 leads to 6)-beta-GlcNAc-1 leads to OC6H4NO2-p, was developed for the rapid identification of the linkage specificity of exo-beta-D-galactosidases from any source via a coupled reaction with N-acetyl-beta-D-hexosaminidase.

Zentralbl Bakteriol {B}, 1980 Mar, 170(3-4), 297 - 303
{Bacteriological and hygienic control of rendering plants (author's transl)}; Thiemann G et al.; Procedures for the hygienic inspection of animal rendering plants should include the bacteriological investigation of the meat meal, the bacteriological control of the used methods of disinfection of the contaminated rooms, of the transport vehicles and of the sewage of the contaminated side . Meat meal has to be free of Clostridium perfringens and Salmonellae . Clostridium perfringens is an indicator of the sterilizing effect of the rendering procedures . After an effective disinfection of rooms no gramnegative and only a small numbers of grampositive bacteria like aerobic bacilli should be demonstrable . Sewage of the contaminated side has to be free of Clostridium perfringens spores.

Can J Microbiol, 1980 Mar, 26(3), 324 - 9
Menadione reductase from Clostridium tyrobutyricum; Petitdemange H et al.; Menadione reductase (EC 1.6.99.2) has been purified 67-fold from Clostridium tyrobutyricum extracts . The molecular weight was found to be 60 000 and the prosthetic group was identified as FMN on the basis of the enzymatic analysis data . The binding of FMN to the menadione dehydrogenase apoenzyme was relatively weak with an apparent Km value of 2.5 x 10(-6) M . The enzyme exhibited group substrate specificity for compounds with a quinoid structure; naphthoquinones and benzoquinones without long carbon chain substituents were the most active . The reactivity of the enzyme with vitamin K1, coenzyme Q6, and cytochrome c was negligible and, with 2,6-dichlorophenol indophenol, relatively low . It was shown that the enzymatic reduction of menadione with the participation of either NADH or NADPH takes place by a "Ping-Pong" mechanism . The enzyme catalyzed the oxidation of NADH and NADPH at equal rates and was inhibited by dicumarol and p-chloromercuribenzoate.

J Pathol, 1980 Mar, 130(3), 193 - 200
The haemorrhagic exudate and its possible relationship to neurogenic inflammation; Malucelli BE et al.; The morphological effects of an aqueous solution of Nistatin, of Clostridium septicum and Tityus serrulatus toxins, of Bothrops jararaca and Agkistrodon piscivorus venoms on the vessels of the tendinous portion of the diaphragm were investigated in guinea-pigs . It was demonstrated that all these substances, when injected intrapleurally induced not only an increase in the permeability of venules but also haemorrhages originating at this segment of the microcirculation . Red cells were shown to escape from venules either by diapedesis or from restricted areas of these vessels which showed intense ultrastructural disorganisation of the vessel wall . Previous treatment of the animals with steroidal or non-steroidal anti-inflammatory drugs or with anticoagulants and chemotherapeutic drugs did not abolish the effect of these irritants . Electrical or chemical stimulation of the phrenic nerve had effects on the microcirculation of the diaphragm similar to those of Nistatin, toxins and venoms . Because of these findings and because Nistatin when injected intrapleurally induces an intense perineuritis of the phrenic nerve, a tentative hypothesis is proposed linking haemorrhagic exudation to the antidromic stimulation of sensitive nerves.

Arch Microbiol, 1980 Mar, 125(1-2), 159 - 65
Utilization of (E)-2-butenoate (crotonate) by Clostridium kluyveri and some other Clostridium species; Bader J et al.; Clostridium La 1 obtained from a Clostridium kluyveri culture was compared with a typical C . kluyvery strain (DSM 555) . The former grows on cortonate and is unable to use ethanol-acetate as carbon sources . The latter grows on crotonate only after long adaptation periods . Resting cells of both strains show also pronounced differences in the fermentation of crotonate . This holds even for C . kluyveri grown on crotonate . Besides several other differences the most striking is that there is no hybridization between the DNA of both strains . Crotonate seems not to be a very special carbon source since C . butyricum and C . pasteurianum grow on crotonate medium supplemented by peptone and yeast extract.

Mol Biol (Mosk), 1980 Mar-Apr, 14(2), 287 - 98
{Conditions for effective hydrogen photoevolution by chloroplasts in the presence of bacterial hydrogenase}; Krasnovskii AA et al.; The hydrogen photoevolution was studied to compare the efficiency of chloroplasts or solubilized chlorophyll in the presence of hydrogenase from Clostridium butyricum and methylviologen which links the electron transfer from photosystems to the exogenous enzyme . The hydrogen evolution by chloroplasts in the absence of exogeneous electron donors (or in the presence of irreversibly oxidized dithiotreitol or cysteine) is probably limited by cyclic electron flow shot-circuiting the photosystem 1 . Efficiency of hydrogen photoproduction when ascorbate or NADP.H are used as electron donors is probably limited by reverse reaction of photoreduced methylviologen with the oxidized electron donor . The combination of both dithiotreitol and ascorbate prevents the shot-circuiting of photosystem 1 by methylviologen; in this case the maximum efficiency of hydrogen photoevolution was achieved up to 400 mumol H2 per 1 mg chlorophyll per hour.

J Clin Microbiol, 1980 Mar, 11(3), 274 - 7
Rapid presumptive identification of anaerobes in blood cultures by gas-liquid chromatography; Sondag JE et al.; Production of volatile and nonvolatile metabolic acids in blood culture broths by aerobic, facultative anaerobic, and obligate anaerobic bacteria was analyzed by gas-liquid chromatography . Anaerobic blood culture isolates were presumptively identified by the qualitative analysis of volatile fatty acids . Isolates, with a characteristic Gram stain reaction and cellular morphology, were identified by the following acid patterns: Bacteriodes fragilis group with acetic and propionic acids; Fusobacterium with acetic, butyric, and usually propionic acids; Veillonella with acetic and propionic acids; gram-positive cocci with acetic and butyric acids; and Clostridium with acetic and butyric acids.

Am J Vet Res, 1980 Mar, 41(3), 348 - 50
Experimentally induced toxicoinfectious botulism in horses and foals; Swerczek TW; Four experiments were performed to elucidate the pathogenesis of toxicoinfectious botulism in horses and foals . Groups of horses and foals were inoculated with one of the following: (1) crude toxin of Clostridium botulinum, type B, given IV, (2) C botulinum spores, given IM, (3) C botulinum spores, given IM, in necrotic lesions, and (4) C botulinum spores, given orally with and without dexamethasone . Toxin of C botulinum in minute amounts is toxic to horses . Clostridium botulinum spores produced toxicosis only when necrotic lesions were present . When C botulinum spores were given orally, they were innocuous . Toxicosis occurred when dexamethasone and C botulinum spores were given orally to a foal with necrotic lesions . Corticosteroids appear to predispose foals to the disease . In the animals where C botulinum organisms infected necrotic lesions, the clinical signs and the lesions seen on necropsy were identical with those seen in spontaneously occurring toxicoinfectious botulism.

J Bacteriol, 1980 Mar, 141(3), 1230 - 8
In vivo and in vitro kinetics of nitrogenase; Davis LC et al.; We measured some of the kinetic parameters of nitrogenase to intact systems of Clostridium pasteurianum and Klebsiella pneumoniae to compare them with the kinetics of the enzyme in vitro . We found that the enzyme showed multiple apparent Km values for acetylene reduction in vivo, as it does in vitro . Carbon monoxide was a noncompetitive inhibitor of acetylene reduction; azide was a noncompetitive inhibitor of acetylene reduction, and nitrogen was a partial inhibitor of acetylene reduction . Cyanide was a noncompetitive inhibitor of acetylene reduction in C . pasteurianum but it was a metabolic poison in K . pneumoniae, in addition to being an inhibitor of nitrogenase . The partial nature of nitrogen inhibition was apparent in assays where both nitrogen and CO were present . Nitrogen did not alter the apparent Ki for CO, nor did the presence of CO enhance the competitive effectiveness of nitrogen . By using recombined nitrogenase fractions, we found that the ability of nitrogen to inhibit hydrogen evolution or acetylene reduction varied with the ratio of protein components . The in vivo inhibition of acetylene reduction by dinitrogen was comparable to that obtained with an excess of the Fe protein in vitro . We conclude that there is an effective excess of the Fe protein available under active growth conditions in vivo.

J Lipid Res, 1980 Mar, 21(3), 381 - 5
3 alpha-, 7 alpha-, and 12 alpha-OH group specific enzymic analysis of biliary bile acids: comparison with gas-liquid chromatograpy; Macdonald IA et al.; 3 alpha-Hydroxysteroid dehydrogenase (3 alpha-HSDH) from P . testosteroni, 7 alpha-HSDH (Escherichia coli ATCC No . 29532) and 12 alpha-HSDH (Clostridium group P, strain C48-50, ATCC No . 29733) were used to directly measure 3 alpha-, 7 alpha-, and 12 alpha-OH groups in extracted human bile-rich duodenal aspirates . Twelve samples chosen from widely differing ratios of cholic/chemodeoxycholic/deoxycholic were computed by solving three simultaneous equations . Comparison of these ratios with those obtained by a) thin-layer chromatography and 3 alpha-, 7 alpha-HSDH assays and b) gas-liquid chromatographic analysis showed no significant difference . Addition of known amounts of pure cholic, chenodeoxycholic, deoxycholic, or lithocholic acid to individual bile extracts gave an appropriate yield of 3 alpha-, 7 alpha-, and 12 alpha-OH groups . The direct (non-chromatographic) enzymic method has the advantages of being rapid, convenient, and inexpensive, and thus suitable for clinical use.

Antimicrob Agents Chemother, 1980 Mar, 17(3), 417 - 22
Bioassay of antibiotics in body fluids from patients receiving cancer chemotherapeutic agents; Wright DN et al.; Patients receiving antitumor chemotherapy are at increased risk of developing nosocomial infections, and the antibacterial therapy of such infections is often monitored by bioassay . The effect of antitumor agents on seven bioassay procedures using strains of Sarcina, Klebsiella, Clostridium, Pseudomonas, Staphylococcus aureus, and S . epidermidis or Bacillus was evaluated . The minimum inhibitory concentrations of six antitumor drugs, cytarabine, dactinomycin, doxorubicin, 5-fluorouracil, methotrexate, and vinblastine, determined for each of the test organisms, showed that 5-fluorouracil, dactinomycin, and doxorubicin are used at blood levels sufficient to interfere with bioassay procedures . The other drugs have minimum inhibitory concentrations as much as 100 times the expected blood levels . Antibiotic (gentamicin, kanamycin, cephalothin, and carbenicillin) recovery experiments in the presence of therapeutic levels of antitumor agents showed no in vitro inactivation of antibiotic . However, at low cephalothin concentrations (less than 20 microgram/ml) in the presnce of 5-fluorouracil, bioassay results were in error by as much as 100% . The data indicate that bioassay procedures for the determining of antibacterial drug levels may need to be modified for those patients receiving antitumor therapy with 5-fluorouracil, doxorubicin, or dactinomycin.

Zentralbl Bakteriol {B}, 1980 Mar, 170(3-4), 252 - 7
{Rendering of animal material in the Netherlands (author's transl)}; Edel W et al.; Just before the World War II rendering of all material of animal origin unfit for human and animal consumption was a fact and was later on regulated by Decree in 1942 . The rendering act dates from 1957 referring to as "processing into useful product" . There is a compulsery rendering which means that withdrawal of the material mentioned from rendering is in general prohibited . Material for rendering must be notified . In use is the dry rendering system (atmospheric batch cookers) . At the moment there are four (three large and one small) rendering plants, the first being established in 1926, and they are under strict government supervision but not under government management . A licence is needed . Supervision is carried out by the Veterinary Public Health Inspectorate . After heating no micro-organism must be present and therefore regular checks for Clostridium perfringens as an indicator organism are carried out besides a contol of ther thermocharts . Recontamination should be avoided . As a result of the extensive bacteriological control since 1972 (Salmonella and other enterobacteriaceae, the latter as indicator organisms) there has been noticed a still growing improvement of the hygienic condition in the rendering plants . During the past half year Salmonella could not be isolated from samples endproduct . Transportation of raw material for rendering after notification to the head of the municipal meat inspection service is done by the rendering plant in completely watertight vehicles . There is a Rendering Board which must be consulted when legislative measures related to rendering are taken.

Appl Environ Microbiol, 1980 Mar, 39(3), 525 - 9
Reversal of radiation-dependent heat sensitization of Clostridium perfringens spores; Gomez RF et al.; The effect of solute concentration on the sensitization of Clostridium perfringens spores to heat by ionizing radiation was investigated . As we have shown previously, spores of C . perfringens treated with gamma radiation are now sensitive to subsequent heat treatments than are spores that receive no radiation treatment . When gamma-irradiated spores were heated in the presence of increasing concentrations of glycerol or sucrose, the heat sensitivity induced by irradiation was progressively decreased . The magnitude of the increase in heat resistance induced by extracellular solutes was greater in gamma-irradiated spores than in nonirradiated spores . Based on these observations, it is proposed that the induction of heat sensitivity in spores by radiation is related to the loss of osmoregulatory or dehydrating mechanisms in irradiated spores.

Ann Microbiol (Paris), 1980 Mar-Apr, 131A(2), 171 - 9
{Transferable tetracycline resistance in "Clostridium difficile" (author's transl)}; Ionesco H; Tetracycline (Tc) resistance is transferable from a resistant strain of Clostridium difficile to a sensitive strain and this resistance is not curable . Resistances to erythromycin and clindamycin are curable but not transferable . These results suggest for these resistances a plasmid determinism . It is shown that a plasmid-mediated Tc resistance (pIP401) of C . perfringens is also transferable to C . difficile . Tc resistance is inducible in C . perfringens and constitutively expressed in C . difficile . In the Tc-resistant transcipients of C . difficile this resistance is either inducible or constitutive, whether the Tc-resistant donor strain was C . perfringens or C . difficile.

Proc Natl Acad Sci U S A, 1980 Mar, 77(3), 1403 - 7
Selenium-containing tRNAs from Clostridium sticklandii: cochromatography of one species with L-prolyl-tRNA; Chen CS et al.; 75Se-Labeled tRNAs were synthesized by Clostridium sticklandii cultures supplemented with 1 microM sodium {75Se}selenite or {75Se}selenocysteine . This process is highly specific for selenium; it occurred in the presence of 1.2 mM sodium sulfide and was not decreased by the further addition of a 500-fold molar excess of cysteine . The 75Se in these tRNAs was located in the polynucleotide portion of the molecules and not in esterified (alkali-labile) selenocysteine . Inhibition of cell multiplication by antibiotics that block either protein synthesis or DNA-dependent RNA synthesis did not prevent this 75Se incorporation . Three {75Se}tRNAs were separated from C . sticklandii cells labeled in the presence of chloramphenicol and were partially purified by chromatography on benzoylated DEAE-cellulose and DEAE-Sephadex A-50 columns . These were designated seleno-tRNAs I, II, and III according to their elution sequence from benzoylated DEAE-cellulose . Cochromatography of purified seleno-tRNA II on DEAE-Sephadex A-50 with an L-proline-accepting species suggests that it is a selenium-containing L-prolyl-tRNA.

Lancet, 1980 Feb 23, 1(8165), 383 - 4
Clostridium difficile associated diarrhoea: a role in inflammatory bowel disease?
Bolton RP, Sherriff RJ, Read AE.
56 patients with diarrhoea were screened for the presence of Clostridium difficile toxin in their stool . The test was positive in 9: 5 had severe inflammatory bowel disease and were receiving systemic steroids; 2 were on steroids for other conditions; 1 had been on antibiotics; and in 1 there was no apparent predisposing factor . In each case clearance of the toxin was associated with clinical improvement . Evidently Cl . difficile toxin is not specific for antibiotic-associated diarrhoea, but is associated with diarrhoea of various aetiologies, often unrelated to antibiotic therapy . Cl . difficile may be an important factor in some exacerbations of inflammatory bowel disease.

Lancet, 1980 Feb 23, 1(8165), 381 - 3
Therapeutic implications of Clostridium difficile toxin during relapse of chronic inflammatory bowel disease; LaMont JT et al.; Clostridium difficile toxin was present in the stools of six patients with chronic inflammatory bowel disease during symptomatic relapse . Only two of these individuals had received antibiotics known to cause pseudomembranous colitis, and on proctoscopy none had pseudomembranes . In all patients disappearance of toxin, either with vancomycin therapy (five patients) or spontaneously (one patient), was associated with symptomatic improvement . Cl . difficile toxin may complicate chronic inflammatory bowel disease, and contribute to relapse in some patients.

Zentralbl Bakteriol A, 1980 Feb, 246(1), 80 - 97
{Investigations on the characterization of a clostridial strain for tumour diagnosis (author's transl)}; Schau HP et al.; Careful delineation of a clostrial strain for human tumour diagnosis as against oncolytic tissue-active clostridial strains should be obligatory . We investigated the possibility to differentiate Clostridium butyricum CNRZ 528 (Bergere) against three different strains kept in our institute by means of biological, biochemical and by phage typing procedures . In addition, consistency in the control of half-technological spore production is required . It could be confirmed by this study that oncolytic properties of clostridial strains are biologically testable with adequate sensitivity in the hamster A Mel 3 tumour model . By means of typing procedures, we were able to clearly differentiate between Clostridium butyricum CNRZ 528 on one hand and Clostridium oncolyticum M 55 as well as Clostridium butyricum Jena H 8 on the other . The results of the tests are unsuitable for identifying Clostridium butyricum CNRZ 528 or Clostridium butyricum McClung 1672 A, respectively, nor can we distinguish Clostridium oncolyticum M 55 from Clostridium butyricum Jena H 8.

J Chir (Paris), 1980 Feb, 117(2), 121 - 3
{Gangrenous cholecystitis without lithiasis . A report on a case diagnosed by simple radiography of the abdomen (author's transl)}; Vinard JL et al.; The authors report a case of gangrenous cholecystitis without lithiasis but with secondary infection of the gallbladder by anaerobic germs . Pre-operative diagnosis was possible from simple x-rays of the abdomen which demonstrated an obvious fluid level in the gallbladder, and by parietography . An immediate cholecystectomy was performed . Bacteriological examination of the bile confirmed the presence of Clostridium perfringens . The main bile duct appeared normal and the bile in the common bile duct was sterile . Convalescence was uneventful with antibiotic cover, in spite of the age and condition of the patient . Gangrenous cholecystitis occurs more frequently without gallbladder stones in humans . Clinical signs are not specific but simple x-rays of the abdomen are pathognomonic and enable pre-operative diagnosis to be made . Urgent cholecystectomy appears to be the ideal treatment . This affection has a high mortality and early operation is therefore justified.

Infect Immun, 1980 Feb, 27(2), 387 - 90
Cecal toxin(s) from guinea pigs with clindamycin-associated colitis, neutralized by Clostridium sordellii antitoxin; Rehg JE; The cecal contents of guinea pigs with clindamycin-associated colitis contained a heat-labile toxin . This toxin was lethal for guinea pigs and mice, produced vascular permeability in the skin of rabbits, and was cytotoxic in tissue culture . The lethality in mice, vascular permeability in rabbit skin, and cytotoxicity in tissue culture monolayers were neutralized by Clostridium sordellii antitoxin.

Jpn J Med Sci Biol, 1980 Feb, 33(1), 1 - 6
Prevalence of Clostridium botulinum in fishes from markets in Osaka; Haq I et al.; A total of 142 samples of different sea foods, mostly fish, were procured from the near-by supermarkets to examine the edible parts for the presence of Clostridium botulinum . Eleven samples (7.7%) seemed to contain this organism . Of these samples, we identified the toxin type in seven; six were type C and the other one type D . Isolation of C . botulinum type C was successful form the six samples but that of type D failed.

J Am Vet Med Assoc, 1980 Feb 1, 176(3), 217 - 20
Toxicoinfectious botulism in foals and adult horses; Swerczek TW; Toxicoinfectious botulism was proved to be the cause of a neuromuscular paralytic syndrome in foals and adult horses . In eight successive cases, Clostridium botulinum type B was isolated at necropsy . Foals were either found dead without premonitory signs of illness or, most often, they had signs of progressive and symmetric motor paralysis . Stilted gait, muscular tremors, and the inability to stand longer than 4 to 5 minutes were the salient clinical signs . Other clinical manifestations included dysphagia, constipation, mydriasis, and frequent urination . As the disease progressed, dyspnea with extension of the head and neck, tachycardia, and respiratory arrest occurred . Death occurred most often 24 to 72 hours after the onset of clinical signs . The most consistent postmortem findings were congestion and edema of the lungs and excessive pericardial fluid, which contained free-floating strands of fibrin . Gastric ulcers, foci of necrosis in the liver, abscesses in the navel and lungs, and wounds of the skin and muscle were predisposing sites for development of toxicoinfectious botulism.

Antimicrob Agents Chemother, 1980 Feb, 17(2), 129 - 31
Comparative in vitro activity of 1-oxa-beta-lactam (LY127935) and cefoperazone with other beta-lactam antibiotics against anaerobic bacteria; Borobio MV et al.; The in vitro activity of 1-oxa-beta-lactam (LY127935), cefoperazone (T-1551), cefuroxime, cefsulodin, cefaclor, cefotaxime, and cefoxitin on 85 anaerobic clinical isolates (30 Bacteroides, 30 Clostridium, 25 Peptococcaceae) was simultaneously determined by the agar dilution test in two different media, Brucella Agar (Difco Laboratories) and Wilkins-Chalgren agar . In Wilkins-Chalgren agar, 90% of Bacteroides were inhibited by (micrograms per milliliter): LY127935, 0.5; T-1551, 64; cefoxitin or cefuroxime, 8; cefsulodin or cefotaxime, 32; and cefaclor, 128 . All Clostridia were inhibited in Wilkins-Chalgren by (micrograms per milliliter): LY127935, 4; T-1551, 2; cefoxitin, 6; cefuroxime, 0.12; cefsulodin, 0.5; cefaclor, 1; and cefotaxime, 8 . All Peptococccaceae were inhibited by T-1551, cefsulodin or cefotaxime at 4 microgram/ml and by cefoxitin or cefuroxime at 1 to 2 microgram/ml . With cefaclor at 8 microgram/ml, 92% of strains were inhibited, and LY127935 at 16 microgram/ml only inhibited 64% of strains . LY127935 was the most active of the antibiotics tested against Bacteroides, showing good activity against Clostridia and poor activity on Peptococcaceae, whereas T-1551 was more active against Peptococccaceae and had similar activity against Clostridia and poor activity on Bacteroides . There are no significant differences between minimal inhibitory concentrations obtained in Brucella Agar and those obtained in Wilkins-Chalgren.

J Infect Dis, 1980 Feb, 141(2), 218 - 22
Effects of Clostridium difficile toxin on tissue-cultured cells; Donta ST et al.; A partially purified toxin of Clostridium difficile induced similar morphologic changes in three different tissue-cultured mammalian cell lines . The morphologic changes were not associated with biochemical changes indentical to those caused by the enterotoxins of Vibrio cholerae and Escherichia coli . Although the mechanisms responsible for the noncytotoxic morphologic effects remain to be delineated, the toxin appears to exert its effects by directly affecting membrane constituents.

J Hyg (Lond), 1980 Feb, 84(1), 151 - 8
Comparison of media and methods for counting Clostridium perfringens in poultry meat and further-processed products; Adams BW et al.; A Most Probable Number (MPN) method involving Differential Reinforced Clostridial Medium followed by streaking on Willis & Hobbs medium was compared with direct plating of samples on Tryptose-Sulphite-Cycloserine agar without egg yolk, and two forms of Oleandomycin-Polymyxin-Sulphadiazine-Perfringens agar, one being prepared from a commercial, dehydrated product . With skin samples taken from chicken carcasses at different stages of processing, the three direct plating media gave similar counts of Cl . perfringens whereas results obtained with the MPN method were consistently lower . Although counts of Cl . perfringens from various further processed products were usually less than 10/g, the three plating media showed similar specificity for this organism . All media supported good growth of reference strains of Clostridium perfringens but it was found that physiologically similar species, including Cl . absonum, Cl . paraperfringens and Cl . perenne also grew uninhibited in these media and produced colonies identical with those of Cl . perfringens, thus indicating the need for confirmatory tests for Cl . perfringens when examining natural samples.

J Biol Chem, 1980 Jan 25, 255(2), 632 - 7
Riboflavin synthases of Bacillus subtilis . Purification and properties; Bacher A et al.; A variety of Bacillus and Clostridium strains were found to contain two forms of riboflavin synthase which can be easily separated by density gradient centrifugation . The fast sedimenting species accounts for 12 to 44% of the total riboflavin synthase activity in the strains analyzed . Both riboflavin synthases were purified to apparent homogeneity from cell extracts of a genetically derepressed mutant of Bacillus subtilis . The specific activities of the pure proteins were 50,000 nmol mg-1 h-1 (light enzyme) and 2,000 nmol mg-1 h-1 (heavy enzyme) . The sedimentation velocities (S20,w) were 4.1 and 26.5 S, respectively . Light riboflavin synthase showed a molecular weight of 70,000 in sedimentation equilibrium experiments . Sodium dodecyl sulfate polyacrylamide gel electrophoresis showed a single band corresponding to a molecular weight of about 23,500 . Thus the enzyme appears to consist of three identical subunits (alpha type) . Heavy riboflavin synthase has a molecular weight of 1,000,000 as shown by sedimentation equilibrium analysis . The protein appears to consist of 2 or 3 alpha subunits and approximately 60 beta subunits . A fragment apparently identical with light riboflavin synthase can be obtained from the heavy enzyme by mild dissociating treatment.

Biochim Biophys Acta, 1980 Jan 25, 595(2), 264 - 76
Mechanism of action of Clostridium perfringens enterotoxin . Effects on membrane permeability and amino acid transport in primary cultures of adult rat hepatocytes; Giger O et al.; Purified enterotoxin from the bacterium Clostridium perfringens rapidly decreased the hormonally induced uptake of alpha-aminoisobutyric acid in primary cultures of adult rat hepatocytes . At 5 min after toxin addition the decrease in alpha-aminoisobutyric acid uptake appeared not due to increased passive permeation (estimated with L-glucose) or to increased alpha-aminoisobutyric acid efflux . When short uptake assay times were employed a depression of alpha-aminoisobutyric acid influx was observed in toxin-treated hepatocytes . The depression of alpha-aminoisobutyric acid influx was correlated with a rapid increase in intracellular Na+ (estimated using 22Na+) apparently effected by membrane damage . In contrast, the uptake of cycloleucine in the presence of unlabeled alpha-aminoisobutyric acid (assay for Na+-independent amino acid uptake) by hepatocytes treated with toxin for 5 min was decreased to only a small extent or not at all depending upon experimental design . At later times, C . perfringens enterotoxin increased the exodus of L-glucose, 3-O-methylglucose and alpha-aminoisobutyric acid from pre-loaded cells indicating that the toxin effects progressive membrane damage . When enterotoxin was removed by repeated washing after 5--20 min the decay of alpha-aminoisobutyric acid uptake ceased and appeared to undergo recovery towards the hormonally induced control level . The degree of recovery of alpha-aminoisobutyric acid uptake was inverse to the length of time of exposure to toxin . Adding at 10 min specific rabbit antiserum against C . perfringens enterotoxin without medium change also reversed the effect of toxin on increased intracellular 22Na+, and on the exodus (from preloaded cells) of alpha-aminoisobutyric acid, L-glucose, and 3-O-methylglucose.

C R Seances Acad Sci D, 1980 Jan 7, 290(1), 41 - 4
{Degradation of collagen by the cariogenic bacteria, Streptococcus mutans}; Despres S et al.; The behaviour of cariogenic Bacteria (Streptococcus mutans) is studied with regard to collagen, which represents 90% of the dentine organic matrix . Collagenase activity of cariogenic Bacteria is measured with radioactive precursors and gel electrophoresis and compared to reference bacterial collagenase (Clostridium histolyticum) . Labelled collagen substrate has been prepared with two different methods: extraction by 0,5 M acetic acid from young Rat skin, previously labelled with L-proline 14C, or reduction by Na B3H4 . Both collagen sutstrates have been incubated for 2 h in Terleckyj medium in which the Streptococcus mutans have been inoculated . The experiments show a proteolytic activity of Streptococcus Mutans on the collagen substrate.

Biochim Biophys Acta, 1980 Jan 4, 589(1), 1 - 9
Biological activity of synthetic molybdenum-iron-sulphur, iron-sulphur and iron-selenium analogues of ferredoxin-type centres; Adams MW et al.; The molybdenum-iron-sulphur cluster {Fe6Mo2S8(SCH2CH2OH)9}3-, which contains two Fe3MoS4 cubane-like centres, is the best plausible analogue available to date for the molybdenum site of the nitrogenase enzymes . The iron-sulphur cluster {Fe4S4(S . CH2CH2OH)4}2- and the iron-selenium cluster {Fe4Se4(S . CH2CH2OH)4}2- are structural analogues of the ferredoxin Fe4S4 active centre . All three clusters would replace ferredoxin and mediate electron transfer to Clostridium pasteurianum hydrogenase in a H2-evolving system with sodium dithionite as the electron donor . The clusters would not replace hydrogenases which themselves are unable to evolve H2 from reduced ferredoxins . The molybdenum-iron-sulphur cluster would also replace ferredoxin in a chloroplast-ferredoxin-hydrogenase H2 evolving system.

Zentralbl Bakteriol A, 1980, 247(4), 495 - 501
Fermentation of 1,2-O-iso-propylidene-D-glucofuranose ("monoacetone glucose") by anaerobic bacteria; Cmelik SH; Various species of Clostridium, Bacteroides, Propionibacterium and Eubacterium were incubated in a 1% solution of 1,2-O-iso-propylidene-D-glucofuranose in a peptone-yeast-extract (PY) medium according to the VPI-technique . The volatile and non volatile acids were investigated by gas-liquid chromatography . All microorganisms showed a pattern of VFA different from that one produced by the glucose containing medium . In most microorganisms the formation of acetic acid was suppressed while the production of propionic, butyric, valeric and iso-valeric acids was stimulated . The production of succinic acid was not affected . Simultaneous determination of monoacetone glucose and glucose in the culture medium showed that the glucose analogue is used to a lesser extent than glucose.

Arch Geschwulstforsch, 1980, 50(1), 53 - 7
{Host range testing of tumour-selective strains of Clostridium butyricum by Bacteriophage 5 (author's transl)}; Schlechte H et al.; Subcultures of Clostridium oncolyticum M55 and in a slightly modified manner of an oncolytic strain of Cl . butyricum H8 are differentiated by lysotypic reaction of the bacteriophage 5 from several tumour-selective non-oncolytic strains and subcultures of Cl . butyricum.

Oncology, 1980, 37(4), 289 - 96
The exposure to humans to nitrite; Walters CL; Although nitrate is more abundant than nitrite in food and the environment in general, it requires reduction by, for instance, bacterial or plant enzymes before it is involved in the nitrosation of amines or amides . Part of the exposure of humans to nitrite arises from its use as a food additive where it performs a very useful function in protecting the consumer from pathogenic micro-organisms such as Clostridium botulinum . Some untreated foodstuffs, such as potatoes, tomatoes and beets, also contain low levels of nitrite . Nevertheless, the main source of human contact is that produced in vivo from nitrate ingested in foods in general and in vegetables in particular . Nitrate also occurs widely in drinking water supplies and this source can also contribute in some measure to human exposure . As yet, it is not possible to compute with accuracy the contribution from any endogenous synthesis within the gastrointestinal tract . Since the rate of nitrosation of an amine is dependent on the nitrite concentration to a power of greater than unity, it is probable that nitrite ingested in one application over a short period will be more active in the synthesis of N-nitroso compounds than a continuous supply at lower concentrations over long periods.

Microbiol Immunol, 1980, 24(5), 393 - 400
Morphological changes during conversion of Clostridium saccharoperbutylacetonicum to protoplasts by sucrose-induced autolysis; Ogata S et al.; When exponentially growing cells of Clostridium saccharoperbutylacetonicum (ATCC 13564) were exposed to hypertonic concentrations of sucrose (0.3--0.5 M), rapid degradation of the cell wall occurred (sucrose-induced autolysis) . The morphological changes from the original rod-shaped cells to protoplasts during the sucrose-induced autolysis were investigated by phase contrast and electron microscopy . When the cells were autolysed in the sucrose solution (0.35 M), each cell began to swell at the middle or at one pole and then formed a small bulb at the swollen part . The bulk consisted of the cytoplasm which was enveloped by the plasma membrane and extruded from the small gap produced by the degradation of the cell wall . The bulb gradually enlarged as lysis progressed, and finally became a protoplast which had no cell wall . The large pre-division cell frequently formed the bulb at the middle (septal site), while the small post-division cell formed the bulb at the pole.

Hoppe Seylers Z Physiol Chem, 1980, 361(6), 875 - 84
Nicotinic acid metabolism enzymic preparation and absolute configuration of the substrate for 2,3-dimethylmalate lyase; Lill U et al.; 1) A convenient method for the enzymatic preparation of a chemically and optically pure isomer of 2,3-dimethylmalic acid in g-amounts is described . Propionate, pyruvate and partially purified 2,3-dimethylmalate lyase (from Clostridium barkeri) were applied . 2) The enzymically formed product, m.p . 99--100 degrees C, {alpha}D20 = -16.4 (water), is related to the known stereochemistry of the Senecio alkaloid jacobine and to a laevorotatory 2,3-dimethylmalic acid derived from jaconecic acid, a degradation product of the alkaloid . From this relationship it appears likely that the substrate of the lyase is a component of the threo racemate and is of (2R,3S) configuration . 3) A three-dimensional X-ray structure analysis was performed and the structure refined to an R value of 0.049 . The asymmetric unit contains three independent threo dimethylmalic acid molecules . The anomalous dispersion effects of carbon and oxygen were used to determine the absolute configuration . These measurements yielded a (2R,3S) configuration . 4) We conclude from these results that (2R,3S)-2,3-dimethylmalate is the substrate of the lyase . The results also establish that previously isolated racemic 2,3-dimethylmalic acids, m.p . 143 degrees C and m.p . 104--106 degrees C, represent the erythro and threo pair, respectively.

Postgrad Med J, 1980 Jan, 56(651), 65 - 6
Clostridium difficile isolated from the stool of a patient with pseudomembranous colits following ampicillin plus flucloxacillin (Magnapen) therapy; Morgan RJ et al.; A case is reported of the isolation of Clostridium difficile from the stool of a patient with antibiotic-related pseudomembranous colitis.

Arch Microbiol, 1980 Jan, 124(1), 111 - 4
Degradation of pyrimidine bases in Clostridium sticklandii; Schafer R et al.; Resting cells of Clostridium sticklandii took up thymine or uracil, when grown in a medium containing 40 mM serine and 20 mM thymine or uracil . The uptake was much lower, when the cells had been grown in a complex medium . Cell-free extracts from cells grown in the complex medium reduced the two bases to the dihydro compounds and decomposed dihydrothymine to beta-ureidoisobutyrate, as indicated by thin-layer chromatography . Uptake and degradation were stimulated by both NADH and NADPH . Further breakdown did not occur, as 14CO2 was not evolved from C-2-labelled thymine or uracil . The rates of pyrimidine uptake and breakdown of C . sticklandii were lower than those reported for C . sporogenes (Hilton et al., 1975).

J Infect Dis, 1980 Jan, 141(1), 92 - 7
Binding of Clostridium difficile cytotoxin and vancomycin by anion-exchange resins; Taylor NS et al.; Cholestyramine and colestipol were tested for binding of Clostridium difficile cytotoxin with use of batch absorption and column chromatography . The toxin was bound by both resins and could not be eluted from cholestyramine with either an ionic of a pH gradient . Vancomycin bound to cholestyramine more strongly than to colestipol . Cholestyramine and vancomycin were also tested for therapeutic efficacy in the hamster model of clindamycin-induced cecitis . Both compounds delayed death and reduced levels of cytotoxin in stool; these effects were greatest for vancomycin . Use of the two compounds in combination reduced concentrations of biologically active vancomycin in stool, but the levels still exceeded the minimum inhibitory concentration for C . difficile . These data suggest that the therapeutic benefit of cholestyramine in some patients with antibiotic-associated pseudomembranous colitis is due to its binding of the C . difficile cytotoxin . Since anion-exchange resins also bind vancomycin, caution is necessary if resins are used concurrently with vancomycin for therapy.

J Bacteriol, 1980 Jan, 141(1), 386 - 8
Influence of growth conditions on glycine reductase of Clostridium sporogenes; Venugopalan V; Cells of Clostridium sporogenes were deficient in glycine reductase activity when grown in a rich medium containing 40 mM each of exogenously added pyruvate and proline or hydroxyproline . These cells lacked the selenoprotein and at least one more protein of the glycine reductase system . Proline or hydroxyproline in the medium also influenced the uptake of glycine by the cells.

Arch Intern Med, 1980 Jan, 140(1), 65 - 8
Pseudobacteremia caused by Clostridium sordellii; Lynch JM et al.; Thirteen of 280 (4.6%) blood cultures collected over a 12-day period were positive for Clostridium sordellii, a spore-forming anaerobe, rarely considered a human pathogen . Nosocomial bacteremia and intrinsic contamination of material used to culture blood were excluded as the source of the organism . Contaminated tincture of thimerosal used to swab the rubber stoppers of blood culture bottles prior to venting (aerobic) or during blind subculturing after 24 hours of incubation (anaerobic) in the clinical microbiology laboratory was determined to be the cause of the pseudobacteremia . After appropriate safe-guards were implemented, we have continued to use tincture of thimerosal for these procedures with no further growth of C sordellii from blood cultures . The importance of less-conspicuous steps in the routine processing of culture material have been reemphasized.

J Foot Surg, 1980 Winter, 19(4), 202 - 6
Gas gangrene: a postoperative complication; Cohen RF et al.; Gas gangrene, also known as clostridial myonecrosis, is a severe and acute infection usually caused by Clostridium septicum, which may contaminate a wound . On rare occasions it is a complication of elective bone surgery, although it is usually found in elderly persons after hip surgery . The onset is usually sudden and it may occur from 6 hr . to 3 days after tissue injury . Diagnosis may be difficult because of the similarity of symptoms to those of anaerobic cellulitis . Treatment consists of surgical debridement, antibiotic therapy, hyperbaric oxygen, and supportive measures . The authors review the literature, discuss the clinical aspects, and present a case history.

Z Erkr Atmungsorgane, 1980, 155(3), 292 - 304
{Application of the serological clostridium assay in tumour diagnostics (author's transl)}; Fabricius EM et al.; The suitability for serologic tumour diagnostics of the non-oncolysing strain Clostridium butyricum CNRZ 528 has been investigated . By introducing the transplantable Brown Pearce tumour of the rabbit and spontaneous tumours of the dog as test models the antibody production against clostridial rods--indicating malignant growth as had been proved with the Mose strain Cl . oncolyticum M55--has been evaluated . It could be established--utilizing the complement fixation test--that one serological method alone is not sufficient for obtaining clear-cut test results in anyone case . It is to be recommended that the indirect hemagglutination and the complement fixation tests ought to be combined . Our studies on specific clostridial antigens and on optimization of techniques will be continued.

Arzneimittelforschung, 1980, 30(7), 1051 - 6
Imidazole derivatives with potential biological activity; Belgodere E et al.; A series of 1-substituted imidazole-5-carbohydroxamic acids Ia, Ib and Ic were prepared from the corresponding 5-methoxycarbonyl imidazoles (IX) obtained by a univocal synthesis starting with the reaction of the amines (III) with ethylchloroacetate . On treatment of 4(5)-methoxycarbonyl imidazoles (XI) with alkylaryl halides (X), on the contrary, mixtures of 1-substituted-4(or 5)-methoxycarbonyl imidazoles were obtained that, when separated by thin-layer chromatography, gave the carbohydroxamic acids Ia, Ib, Id and Ie and IIa leads to f . The structure of the imidazole derivatives were obtained by means of IR, NMR and UV spectra . On carrying out tests of biological activity on these compounds, it had been found that the 5-carbohydroxamic acids possess, compared to the 4-carbohydroxamic ones, a greater activity . Particularly Ib and Ib--HCl seem fairly active against Klebsiella pneumoniae and Clostridium bifermentans, Ib--HCl against Bacillus subtilis, too.

Scand J Infect Dis Suppl, 1980, (Suppl 22), 16 - 29
Interaction of cytopathogenic toxin from Clostridium difficile with cells in tissue culture; Thelestam M et al.; Partially purified cytopathogenic toxin from Clostridium difficile induced morphological changes in five cell lines in tissue culture . The relative sensitivity scale of the cell lines was human lung and intestinal fibroblasts greater than Chinese hamster ovary cells much greater than mouse adrenal cells greater than mouse neuroblastoma cells . The cytopathogenic effect did not occur in toxin-treated lung fibroblasts incubated at 0 degree C . Pre-incubation of lung fibroblasts with 2,4-dinitrophenol prevented the cytopathogenic effect . The toxin bound to as yet unidentified receptors at the surface of human lung and intestinal fibroblasts . The toxin-induced morphological (actinomorphic) changes in lung and intestinal fibroblasts closely resembled the effects induced by the fungal metabolite cytochalasin B (CB), which is known to disrupt microfilaments reversibly . Indirect immunofluorescence with anti-actin antiserum demonstrated that the C . difficile toxin disrupted the straight actin filament bundles seen in normal fibroblasts . The cytopathogenic effect became apparent 3--5 h after exposure to toxin . However, irreversible intoxication occurred already within 20 min of exposure, as toxin-treated fibroblasts which were trypsinized and reseeded were not able to attach to the solid substratum and regenerate their typical shape, a process requiring reorganization of actin into microfilament bundles . Two possible different modes of action of the toxin, leading to microfilament disruption, are suggested: 1) Transmembrane signal by surface-bound toxin via microfilament-linked integral membrane protein(s) and 2) Penetration of surface-bound whole toxin or an active fragment, followed by its intracellular action . The experimental evidence so far is consistent with either of these mechanisms.

Infection, 1980, 8 Suppl 2, S163 - 6
Introduction of anaerobic methodology into a clinical microbiological laboratory; Sonntag HG; For the successful introduction of anaerobic methodology into a clinical microbiological laboratory, several factors are very important . These include: 1) laboratory; 2) establishment of skilful technicians and adequate equipment; and 3) communication with clinicians who believe in anaerobes as infectious agents and are interested in working with the microbiologist . The beneficial effects of these factors on a laboratory's efficiency in providing an anaerobic service are demonstrated by the following data . In the period January to July 1978, we found 234/1446 specimens (16.2%) positive for anaerobes . The isolated strains mainly belonged to the genera Propionibacterium (26.5%), Bacteroides (26.1%), Clostridium (26.1%) and gram-positive cocci (18.4%) . Fusobacterium could be found in 1.7% only . Specimens yielding anaerobes were mostly derived from infected wounds, pus, abscesses, and aspirated liquids from the pelvis, gall bladder or knee . The problem which now has to be solved concerns the significance of some of the anaerobic isolates as causative infectious agents in individual cases.

Infection, 1980, 8 Suppl 2, S117 - 22
The clinical significance, taxonomy and special methodological problems of the pathogenic clostridia; Bittner J; The clinical significance of clostridia is much greater than is generally recognized . The organisms are a major cause of septic abortion, Clostridium perfringens being the most important single organism . This species is also the principle agent in food-poisoning . Clostridium botulinum is considered to be one of the main causes of the sudden death syndrome in infants . As clostridia are universally distributed in nature and the human body, the isolation of an organism of this group from the human body is significant only if it can be linked with pathological changes . In the case of histotoxic disease, a direct gramstained smear from the lesion is of paramount importance, since the pathogens are always present in large numbers . Generally, a few simple procedures and tests ensure the rapid isolation and identification of the main pathogenic clostridia . C . perfringens, by far the most important species, may be identified by its ability to produce lecithinase on egg yolk-glucose agar and stormy clot in litmus milk . However, strain identification of this microorganism is much more complicated.

Microbiol Immunol, 1980, 24(7), 575 - 84
Purification and some properties of tetanolysin; Mitsui N et al.; Tetanolysin was purified from the culture fluid of a strain of Clostridium tetani by ammonium sulfate fractionation, acetone precipitation and repeated gel filtration . Two hemolysins with different molecular weights were separated by gel filtration, and the smaller one, tetanolysin, was further purified . The purification raised the specific activity of tetanolysin 1,050-fold to 500 HU/micrograms of protein . The purified preparation gave a single, relatively broad band on polyacrylamide gel electrophoresis, in which the activity was roughly parallel with the protein concentration . However, on sodium dodecylsulfate-gel electrophoresis it gave two bands with nearly equal amounts of proteins, showing molecular weights of 53,000 and 48,000 +/- 3,000 . Furthermore, isoelectric focusing revealed four peaks of the activity whose isoelectric pHs were 6.1, 5.6, 5.3, and 6.6 in decreasing order of the activity . These findings suggest that the preparation contains four hemolysins with different pIs, which are classifiable into two groups by molecular size . The preparation was completely free of tetanus neurotoxin and proteases . Tetanolysin was more strongly inhibited by cholesterol and more rapidly absorbed onto erythrocytes than theta-toxin of Cl . perfringens.

Microbiol Immunol, 1980, 24(6), 507 - 13
Ultrastructure of a hexagonal array in exosporium of a highly sporogenic mutant of Clostridium botulinum type A revealed by electron microscopy using optical diffraction and filtration; Masuda K et al.; The ultrastructure of a hexagonal array in the exosporium from spores of a highly sporogenic mutant of Clostridium botulinum type A strain 190L was studied by electron microscopy of negatively stained exosporium fragments using optical diffraction and filtration . The exosporium was composed of three or more lamellae showing and equilateral, hexagonal periodicity . Images of the single exosporium layer from which the noise had been filtered optically revealed that the hexagonally arranged, morphological unit of the exosporium was composed of three globular subunits about 2.1 nm in diameter which were arranged at the vertices of an equilateral triangle with sides of about 2.4 nm . The morphological units were arranged with a spacing of about 4.5 nm . the adjacent globular subunits appeared to be interconnected by delicate linkers.

Microbiol Immunol, 1980, 24(6), 469 - 77
Isolation of nontoxigenic variants associated with enhanced sporulation and alteration in the cell wall from Clostridium botulinum type a 190L by treatment with detergents; Takumi K et al.; Nontoxigenic variants were isolated from Clostridium botulinum type A strain 190L after treatment with detergents such as deoxycholate, sodium dodecyl sulfate, Tween 80 and Brij-58 . Deoxycholate was most effective for obtaining the variants . The variants exhibited a markedly increased frequency of sporulation compared with the oligosporogenic parent strain . The cell wall of the parent strain was composed of an outer layer and an inner layer, whereas that of the variants lost the outer layer . After treatment with mitomycin C the parent strain was subjected to lysis and produced bacteriophages with a hexagonal head and a contractible tail, while the nontoxigenic variants did not yield bacteriophages or phage-like structures . There appears to be a close relationship among the toxigenic and sporogenic properties, formation of the outer cell wall layer and lysogeny.

Annu Rev Biochem, 1980, 49, 93 - 110
Selenium-dependent enzymes; Stadtman TC; Selenium, molecular weight 78.96, resembles sulfur in many of its chemical properties and occurs in inorganic forms as H2Se, H2Se2O3, H2SeO3, and H2SeO4 which are the analogues of hydrogen sulfide, thiosulfate, sulfite, and sulfate, respectively . The commonly available radionuclide, 75Se, is a gamma emitter (half-life 122 days) that is used extensively as a tracer in biochemical studies and as a radiopharmaceutical agent for diagnostic purposes . Organoselenium compounds, in general, are less stable and more reactive than the corresponding sulfur analogues and these properties may account for the toxicity of selenium when it is incorporated indiscriminately in place of sulfur in cellular constituents . On the other hand living systems may have exploited the greater reactivity of certain types of organoselenium compounds in those instances where selenium is specifically required as a component of an enzyme or other macromolecule . Several enzymic processes that do not distinguish selenium from sulfur and therefore may be important in selenium toxicity were discussed in some detail in two earlier reviews on selenium biochemistry (1, 2) and this aspect of the problem is not treated here . Rather, the information currently available on the properties and catalytic functions of the four known selenium-dependent enzymes is summarized . These enzymes are formate dehydrogenases of Escherichia coli and several anaerobic bacteria, clostridial glycine reductase, mammalian and avian glutathione peroxidase, and nicotinic acid hydroxylase of Clostridium barkeri . Additional selenoproteins whose catalytic activities are as yet unidentified are mentioned.

Dev Biol Stand, 1980, 45, 143 - 9
The effects of laboratory animal diets on the potency tests of bacterial vaccines; Knight PA et al.; Comparative studies of the responses elicited by mice fed on PCD and FFG diets to a number of bacterial vaccines have shown a significant reduction in the immune response to tetanus toxoid but not to Clostridium septicum toxoid and increased resistance to challenge with E . coli and syngeneic tumour cells but not to Pasteurella multocida . These differences cannot readily be explained in terms of differences between the identifiable constituents of the diets and illustrte the dangers of vaccine potency tests that require an absolute level of response to the material under test.

Microbiol Immunol, 1980, 24(4), 271 - 9
Studies on the sulfite reduction test for clostridia; Kawabata N; Peptone-yeast extract (PY) medium containing 0.035% ferric ammonium citrate as an indicator, 0.05% sulfite as a substrate, 0.05% cysteine as a reducer and 0.5% glucose was found to be suitable for observing the sulfite reduction test . The effect of added cysteine on the test was suppressed by the addition of glucose . In cultures of bacteria grown for 2 days at 37 C in medium containing the above ingredients, 121 among 132 strains of clostridia, including 86 strains of Clostridium perfringens, gave a positive reaction . Although some strains of Salmonella and Proteus were positive, the specificity of the test for clostridia was thought to be relatively high . Positive reactions in a resting cell system were limited to some species of clostridia.

Scand J Infect Dis Suppl, 1980, (Suppl 22), 7 - 10
The experimental pathogenesis of antibiotic related colitis; Larson HE; Findings from several countries now closely associate Clostridium difficile and its toxin with PMC . In fact, testing for the toxin by means of tissue culture assay is being used more and more to define the proportion of patients with clinically significant antibiotic associated colitis . Reproduction of a similar entity in hamsters appears to fulfil the Koch-Henle postulates, establishing C . difficile as the cause of the syndrome . Antibiotic treatment creates susceptibility to infection rather than being directly responsible for the lesions . The manner in which this occurs is not clear . Animal experiments show that C . difficile is present in some environments and that it may spread through the air and be ingested to cause infection.

Scand J Infect Dis Suppl, 1980, (Suppl 22), 37 - 40
Characterization of Clostridium difficile and its differentiation from Clostridium sporogenes by automatic head-space gas chromatography; Larsson L et al.; Although 47 strains of Clostridium difficile and Clostridium sporogenes were studied by gas chromatography . Acidic and neutral volatile products, formed after 96 h of incubation in glucose-containing peptone yeast-extract medium, were chromatographed . All strains produced appreciable amounts of fatty acids, which were tentatively identified by gas chromatographic retention data . Chromatograms obtained when analysing diethyl ether extracts of culture media of all 47 strains were virtually identical . However, analysis of the broth media by automatic head-space gas chromatography, employing a glass capillary column, gave chromatographic patterns which differentiated the two Clostridium species studied . C . sporogenes was characterized by chromatographic patterns containing a dominant peak of isovaleric acid . Strains of this species produced larger amounts of early eluting compounds and much smaller amounts of butyric and valeric acid than did strains of C . difficile . Automatic head-space gas chromatography provides an efficient means for differentiation of C . difficile and C . sporogenes . This gas chromatographic technique is easier and more rapidly performed than analysis of either extracts of culture media.

Scand J Infect Dis Suppl, 1980, (Suppl 22), 11 - 5
Experimental studies of antibiotic associated colitis; Bartlett JG; Clostridium difficile has been implicated as the major cause of antibiotic-associated pseudomembranous colitis . The laboratory diagnostic test of choice is a tissue culture assay that demonstrates the presence of a cytopathic toxin neutralized by antitoxin to Clostridium sordelli . This toxin is found in stools from patients with antibiotic-associated pseudomembranous colitis and in stools from patients with antibiotic-associated diarrhea . Neutralization of toxin by antitoxin to C . sordelli appears to represent antigenic cross-reactivity, since both cultures of C . difficile also contain a cytopathic toxin neutralized by this toxin . Strains of C . difficile are susceptible to vancomycin and the clinical experience with oral administration of this agent shows promising results.

Antonie Van Leeuwenhoek, 1980, 46(6), 523 - 31
New isolation of Clostridium aceticum (Wieringa); Adamse AD; After many attempts to re-isolate Clostridium aceticum (Wieringa) had been unsuccessful, finally a new strain could be isolated that was morphologically and physiologically identical, as could be demonstrated by comparing this strain with the original one, retrieved recently from an old culture collection . Both strains showed the ability to produce cellular materials and acetate from a CO2-H2 gas mixture, as well as from fructose as the substrate . A detailed description of the enrichment and isolation procedures used, is given.

Cytobios, 1980, 29(114), 99 - 108
Association of vegetative antigens with the spores of Clostridium sporogenes; Princewill TJ; Clean spores of Clostridium sporogenes stored for two years continued to elicit the production of specific antibodies in immunized rabbits . Disintegrated spores stimulated not only antibodies specific for the spore component, but also those directed against 'H' and 'O' antigens of the sporangium cells.

Arch Microbiol, 1980 Jan, 124(1), 73 - 9
Methane formation from fructose by syntrophic associations of Acetobacterium woodii and different strains of methanogens; Winter JU et al.; When Acetobacterium woodii was co-cultured in continuous or in stationary culture with Methanobacterium strain AZ, fructose instead of being converted to 3 mol of acetate was converted to 2 mol of acetate and 1 mol each of carbon dioxide and methane, showing that interspecies hydrogen transfer occurred . In continuous culture the organisms formed a close physical association in clumps; the doubling time for each organism was 6 h at 33 degrees C . Methane mainly was derived from carbon positions 3 and 4 of the sugar, but other carbons also yielded methane; this was shown to be due to carbon dioxide-acetate exchange reactions by A . woodii in a manner similar to that carried out by Clostridium thermoaceticum . Four other methanogens, Methanobacterium M.o.H . and M.o.H . G, Methanobacterium formicicum, and Methanosarcina barkeri (not acetate-adapted) also produced similar results, when co-cultured with A . woodii.

Rev Argent Microbiol, 1980 Jan-Apr, 12(1), 10 - 7
{Toxoids of botulinum toxin type G}; Puig de Centorbi ON et al.; Clostridium botulinum type G toxin was obtained by the dialysis sac culture method . Crude toxin was submitted to precipitation either by 4.5 M (NH4)2SO4 (Table 2) or ethanol 96% up to 25% final concentration (Table 3) . Aliquots of crude toxin and fractions from the precipitation methods were activated by trypsin, detoxificated by formalin and adsorbed with aluminum phosphate . Twelve preparations of toxoids (Table 1) were obtained and assayed in laboratory animals . The immune response was studied through the toxin-antitoxin neutralization test, set up at a level of 4,000 mouse LD50 per ml . Guinea pigs had the highest titer of antitoxin (64,000 anti-mouse-LD50/ml) after its immunization with toxoid prepared with toxin precipitated by 4.5 M ammonium sulphate activated by trypsin and adsorbed with aluminum phosphate . Rabbits responded with a lower titer of antitoxin but had a similar response than guinea pigs to the same toxoids (Table 5) . Chickens did not show any antitoxin response above 4,000 anti-mouse-LD50 per ml.

Arch Geschwulstforsch, 1980, 50(7), 601 - 12
A theoretical models of oncolysis by Clostridium oncolyticum M 55 ATCC 13,732; Brantner H et al.; Based on experimental data obtained by own experiments and investigations of other authors the first time it is tried to develop a new theoretical model of oncolysis by Clostridium oncolyticum M 55 . The correlations between tumour, clostridial cells, the tumour kinin system and the immune system of the host are represented in their influence on oncolysis . The consequences for a therapeutic application of the tumour clostridia phenomenon are discussed.

Zentralbl Bakteriol Mikrobiol Hyg {B}, 1980, 171(6), 538 - 43
{The suitability of bioindicators according to DIN 58948 part 4 for monitoring gas-sterilizers (author's transl)}; Adam W et al.; In the Federal Republic of Germany bioindicators according to DIN 58948, Part 4, are generally used for testing the efficacy of ethylene oxide sterilizers . They are based on spores of Bacillus subtilis dried in sheep-blood on the bottom of a small test tube . As some authors doubted the resistance of these indicators to ethylene oxide especially in comparison with spore soil and spores of Clostridium perfringens, three different test procedures were performed showing that spores of sulfite reducing clostridia (Cl . perfringens included) are less resistant to ethylene oxide than spores of B . subtilis . The results are summarized in tables 1 to 3 . They are discussed with regard to literature on the subject with special emphasis to the significance of occlusion of spores in insoluble crystals . It is concluded that spores of B . subtilis are the most suitable test-organisms for monitoring ethylene oxide sterilization and that spore soil cannot be used for this purpose.

Infection, 1980, 8 Suppl 2, S171 - 5
Methods for testing antibiotic sensitivity of anaerobic bacteria; Garcia-Rodriguez JA et al.; Problems are still encountered in the performance and interpretation of tests of anaerobe sensitivity to antibiotics . A review of the methods currently used was carried out in order to determine factors modifying the activity of antibiotics . The sensitivity of Escherichia coli, Staphylococcus aureus and Clostridium perfringens to various drugs was tested under different conditions (including different culture media and incubation atmospheres) . Gentamicin and kanamycin showed no activity in tests with brain heart infusion agar incubated anaerobically or in 10% CO2 . The activity of lincomycin was much more readily influenced by test conditions than that of clindamycin . All the drugs examined except gentamicin and kanamycin showed more activity when tested in brain heart infusion agar than in Mueller Hinton blood agar.

Microbiol Immunol, 1980, 24(7), 595 - 601
Effect of oxidizing agents and sulfhydryl group reagents on beta toxin from Clostridium perfringens type C; Sakurai J et al.; Purified beta toxin from Clostridium perfringens type C was inactivated by the oxidizing agents o-iodosobenzoate (OIBA), oxidized glutathione, and ferricyanide, and by the sulfhydryl group regents 5,5'-dithio-bis(2-nitrobenzoic acid) (DTNB) and N-ethylmaleimide, iodoacetamide, and iodoacetic acid, causing loss of activity in various degrees depending on the concentration used . The activity of the toxin was not influenced by exposure to 1.0 mM of p-chloromercuribenzoate . The toxin treated by OIBA or DTNB was reactivated by incubation with 2-mercaptoethanol and dithiothreitol . The data suggest that beta toxin contains thiol groups which are essential for the activity.

Eur J Biochem, 1980, 107(1), 25 - 30
The ganglioside content of the milk fat-globule membrane and the mouse mammary-tumour virus isolated from the milk of infected mice . Partial characterization of a new disialoganglioside; Gosselin-Rey C et al.; The milk fat-globule membrane and the mouse mammary-tumour virus isolated from the milk of infected Swiss mice have been investigated for their content in gangliosides . When compared on the lipid phosphorus basis, viral envelope is found to contain more than twice as much lipid-bound sialic acid as fat-globule membrane . The ganglioside patterns of these two structures appear rather similar, except for the occurrence in fat-globule membrane of a low ganglioside homolog, presumably GM2, not detected in viral envelope . A common and dominant trait is the presence in both structures, as the main ganglioside, of a component which has been so far characterized as a disialoganglioside, having the same neutral glycolipid moiety as GD1a, but with both sialic acid residues displayig to Clostridium perfringens and Vibrio cholerae neuraminidase, the susceptibility typical of terminal sialic acid residues.

Appl Environ Microbiol, 1980 Jan, 39(1), 118 - 26
Enumeration of potentially pathogenic bacteria from sewage sludges; Dudley DJ et al.; To ascertain the health risks that may be posed by the land application of sewage sludges, a scheme was devised to determine the types and numbers of pathogenic and potentially pathogenic bacteria present in sludges . A processing treatment was adapted to sludge to give a homogenate which yielded the greatest numbers of viable bacteria . Conventional methods were successful in enumerating Klebsiella, Staphylococcus, gram-negative enteric bacteria, and commonly used indicator organisms . Modifications of conventional methods improved the enumeration of Salmonella, Mycobacterium sp., fluorescent Pseudomonas sp., and Clostridium perfringens . However, Shigella methodology yielded only one isolate . Utilizing the proposed scheme, the population densities of these organisms were estimated in three domestic wastewater sludges . In light of these results, the potential impact of land application of sewage sludges is discussed.

Biochim Biophys Acta, 1979 Dec 6, 548(3), 552 - 64
The electron spin relaxation of the electron acceptors of photosystem I reaction centre studied by microwave power saturation; Rupp H et al.; Photosystem I particles from spinach were reduced by illumination at 77 K . Under these conditions the one-electrom transfer from P-700 resulted in a reduction of only one acceptor molecule of the reaction centre . The EPR signals at g=2.05, 1.94 and 1.86 were attributed to reduced centre A and the smaller signals at g=2.07, 1.92 and 1.89 to reduced centre B . Reduction of both centres by dithionite in the dark lead to signals at g=2.05, 1.99, 1.96, 1.94, 1.92 and 1.89 . Thus, the features at g=2.07 and 1.86 disappeared and new signals at g=1.99 and 1.96 were observed . From the spectral changes it followed that the iron-sulphur centres A and B interact magnetically . Temperature dependent EPR spectra demonstrated a faster electron spin relaxation of centre A than of centre B . These conclusions were corroborated using microwave power saturation of the respective EPR signals . The saturation data of the fully reduced centres A and B could not be fitted using the saturation equation for a one-electron spin system . The magnetic interaction between the (4Fe-4S) CENTRes of the electron acceptors A and B resulted in saturation properties which are simular to those of the 2(4Fe-4S) ferredoxin from Clostridium pasteurianum . For centre X a high proportion of homogeneous broadening of the EPR lines was inferred from the inhomogeneity parameter (b=1.83) . It was, therefore, concluded that centre X is most probably an anion radical of chlorophyll . From the low temperature necessary for observing the EPR signal of centre X followed that the drastic relaxation enhancement has to be attributed to a magnetic interaction of the anion radical with iron.

NIPH Ann, 1979 Dec, 2(2), 17 - 24
Nitrite as a food additive; Dahle HK; Nitrite is used for its colouring, antimicrobial and flavouring effects as a food additive for several meat, fish and cheese products . Nitrite combines readily with secondary amines to form carcinogenic nitrosamines . Nitrosamines are found in many food products after nitrite addition and sometimes even without addition . Nitrite is regarded as an effective growth inhibitor for Clostridium botulinum and thereby its production of the lethal toxin . Today this is considered to be the main reason for addition of nitrite to food products . It should be possible to limit the addition of nitrite to a few special food products where Cl botulinum really represents a hazard to human health, i e to canned meat that is not sterilized by heat and some cured and fermented products . In Norway the use of nitrite is limited to products where growth of clostridia is possible, but in a few products nitrite is also allowed as a colour stabilizer . It is reasonable to expect that other countries will decide upon similar regulations . The naturally occurring nitrates in vegetables have to be included in the discussion due to the possibility of microbial reduction to nitrites.

Hoppe Seylers Z Physiol Chem, 1979 Dec, 360(12), 1693 - 702
Nicotinic acid metabolism . 2,3-Dimethylmalate lyase; Pirzer P et al.; 1) A new enzyme, 2,3-dimethylmalate lyase, was purified from Clostridium barkeri to about 80% homogeneity . Some of the properties of the enzyme are described . 2) It is shown that the 2,3-dimethylmalic acid (m.p . 143 degrees C) described in the literature represents only one racemic pair . This pair is not attacked by 2,3-dimethylmalate lyase . 3) The isolation of both racemic pairs of 2,3-dimethylmalic acid is described . Half of one pair, m.p . 104-106 degrees C, was converted to propionate and pyruvate by 2,3-dimethylmalate lyase . 4) In combination with earlier work performed by E.R . Stadtman and coworkers the results given under points 1--3 establish 2,3-dimethylmalate as an intermediate in the degradation of nicotinic acid by C . barkeri . 5) Experimental evidence indicates the 2,3-dimethylmalate lyase is no acyl-S-enzyme and that it is different in this respect as well as in quaternary structure from the apparently related enzymes citrate lyase and citramalate lyase.

Am J Vet Res, 1979 Dec, 40(12), 1752 - 6
Immunogenicity of Clostridium septicum in guinea pigs; Claus KD et al.; Five strains of Clostridium septicum were used to prepare bacterins, bacterin-toxoids, toxoid, and combinations of bacterins or bacterin-toxoids . These preparations were tested for immunogenicity in guinea pigs vaccinated subcutaneously with 1.0 ml of product . Usually, a second vaccination was given 21 to 24 days later . The immunity of groups of vaccinated guinea pigs was challenged with as many as 22 strains of C septicum . When challenge exposed with homologous strains at 21 to 24 days after one vaccination or 10 t0 18 days after a second vaccination, 60% to 100% of the guinea pigs in each group survived . Demonstrable cross-protection among strains of C septicum varied from none to 100% protection in vaccinated guinea pigs . A combination of bacterin-toxoid prepared from four selected strains protected 70% to 100% of the vaccinated guinea pigs challenge exposed with 21 strains . Duration-of-immunity studies demonstrated a twofold to fourfold decrease in protection when the vaccination-to-challenge interval was extended an additional 3 weeks . Strains of C septicum do not have an effective common immunogen and the stimulated immunity appears to be of short duration . Antitoxin was demonstrated to be less important than other factors in protecting against C septicum infection.

J Clin Microbiol, 1979 Dec, 10(6), 880 - 4
Cultures for Clostridium difficile in stools containing a cytotoxin neutralized by Clostridium sordellii antitoxin; Willey SH et al.; Stools from patients with antibiotic-associated diarrhea or colitis were cultured to detect the presence of Clostridium difficile . All specimens contained a cytotoxin which was neutralized by Clostridium sordellii antitoxin . Initial testing employed several methods with comparative merits in recovering this organism . These included the use of nonselective media, antibiotic-incorporated media, alcohol shock, and paracresol-containing broth . Optimal results were achieved with primary plating of serial dilutions onto a selective agar containing cycloserine and cefoxitin . This technique was then employed in a large number of specimens . The overall results showed that C . difficile was recovered in specimens from 71 of 73 patients . All isolates of C . difficile produced a cytotoxin which was neutralized by C . sordellii antitoxin in vitro . These results verify the utility of this medium and support the concept that C . difficile accounts for the cytotoxin found in stools in nearly all cases.

Can J Microbiol, 1979 Dec, 25(12), 1352 - 8
Indigenous bacteria influence the number of Salmonella typhimurium in the ileum of gnotobiotic mice; Roach S et al.; Gnotobiotic BALB/c mice associated with indigenous Lactobacillus, Bacteroides, and two fusiform-shaped Clostridium strains had fewer S . typhimurium present in the ileum 3 days after intragastric challenge with the pathogen than did similarly challenged germfree mice . Acetic and butyric acids were detected in the caecal contents of the gnotobiotic mice, but in smaller concentration than was present in conventionalized mice . No difference in the motility of the small intestine was detected between germfree, gnotobiotic, and conventionalized mice.

Appl Environ Microbiol, 1979 Dec, 38(6), 1081 - 5
Separation of botulinum-positive and -negative fish samples by means of a pattern recognition method applied to headspace gas chromatograms; Snygg BG et al.; A gas chromatographic headspace technique was used to analyze the gas produced during putrefaction of pond-raised, degutted trout, incubated in evacuated plastic pouches . The following samples were analyzed; 10 samples which, due to natural contamination with Clostridium botulinum, were toxic when injected into mice, 10 samples which were nontoxic when injected, and 9 samples inoculated with one strain of C . botulinum type E . The gas chromatograms showed the presence of 118 compounds in most samples . Quantitative differences among most chromatograms could be observed, but no compound was unique to any of the three groups . By means of a specific pattern recognition method, all negative samples were shown to fall into one group and were distinctly separated from the toxic samples . No differences could be observed between the two groups of inoculated and naturally contaminated trout samples . The results suggest that headspace analysis combined with pattern recognition analysis might prove to be a valuable method for screening studies of foods containing living cells of C . botulinum.

Arch Intern Med, 1979 Dec, 139(12), 1346 - 9
Clinical characteristics of anaerobic bactibilia; Bourgault AM et al.; During a two-year period, 1,892 patients underwent biliary tract surgery at the Mayo Clinic . Both aerobic and anaerobic bile cultures were performed in 371 patients and 253 of these were positive . Anaerobes were isolated from 100 patients, although only twice in pure culture . Only aerobes grew from cultures from 153 patients . One hundred cases of biliary tract infections involving anaerobes and an equal number involving aerobes only were reviewed in order to determine their clinical characteristics . Prominent features of anaerobic bactibilia included (1) a history of complex, multiple, biliary tract surgeries often involving biliary-intestinal anastomoses and common bile duct manipulation, (2) severe symptoms, (3) high incidence of postoperative infectious complications, especially wound infections . Further analysis of anaerobic biliary infections suggested that Bacteroides fragilis was more often associated with serious pathologic conditions of the biliary tract than was Clostridium.

Biomedicine, 1979 Dec, 31(9-10), 250 - 2
Effect of Clostridium perfringens neuraminidase on viability and antigenicity of human leukemic myeloblasts; Ogier C et al.; The effect of increasing concentrations of Cl . Perfringens neuraminidase and of pH on the dye exclusion ability and lymphocyte stimulating capacity of leukemic myeloblasts was studied . The higher the neuraminidase concentration, or the lower the pH was, the more myeloblasts died and the less the myeloblasts stimulated lymphocytes . Myeloblasts treated at a neutral pH and at low enzyme concentrations retained, but did not increase their antigenicity.

Vet Rec, 1979 Nov 24, 105(21), 480 - 2
Cerebrocortical necrosis in ruminants: effect of thiaminase type 1-producing Clostridium sporogenes in lambs; Cushnie GH et al.; Large numbers of orally inoculated thiaminase type 1-producing Clostridium sporogenes failed to establish in the alimentary tract of two conventionally born lambs . Conversely, when similar inoculations were given to two gnotobiotic lambs, large populations of Cl sporogenes established in their rumens and correspondingly high levels of thiaminase were produced . No clinical symptoms of thiamine deficiency or cerebrocortical necrosis were seen despite the presence of high levels of thiaminase in the rumen of one of the gnotobiotic lambs for a period of 86 days.

Biochim Biophys Acta, 1979 Nov 16, 558(1), 48 - 57
Asymmetry of the site of choline incorporation into phosphatidylcholine of rat liver microsomes; Higgins JA; {14C}Choline was incorporated into microsomal membranes in vivo, and from CDP-{14C}choline in vitro, and the site of incorporation determined by hydrolysis of the outer leaflet of the membrane bilayer using phospholipase C from Clostridium welchii . Labelled phosphatidylcholine was found to be concentrated in the outer leaflet of the membrane bilayer with a specific activity approximately three times that of the inner leaflet . During incorporation of CDP-choline and treatment with phospholipase C the vesicles retained labelled-protein contents indicating that they remained intact . When the microsomes were opened with taurocholate after incorporation of {14C}choline in vivo, the labelled phosphatidylcholine behaved as a single pool . Selective hydrolysis of labelled phosphatidylcholine in intact vesicles is not, therefore, a consequence of specificity of phospholipase C . These results indicate that the phosphatidylcholine of the outer leaflet of the microsomal membrane bilayer is preferentially labelled by the choline-phosphotransferase pathway and that this pool of phospholipid does not equilibrate with that of the inner leaflet.

J Biol Chem, 1979 Nov 10, 254(21), 10728 - 33
Structure of tetanus toxin . Demonstration and separation of a specific enzyme converting intracellular tetanus toxin to the extracellular form; Helting TB et al.; Protease activity has been demonstrated in culture supernatants of Clostridium tetani at various stages of fermentation . Gel chromatography of the concentrated filtrates revealed the presence of three enzymatically active fractions eluting at separate positions off the column . The smallest protease was found to "nick" the single chain intracellular tetanus toxin, producing the extracellular, two-chain structure of the molecule . As little as 3 ng of active protease were sufficient to cleave 50 microgram of intracellular tetanus toxin, suggesting that this enzyme is responsible for the observed structural change of the toxin molecule during its release into the culture medium . By comparison, the second protease, eluting at an intermediate position, exhibited only marginal activity towards intracellular toxin . The third, largest, enzyme was not active under the conditions of the assay . However, the latter protease effectively hydrolyzed low molecular weight histidyl peptides, and it is concluded that this enzyme is similar to the one described by Miller, P.A . Gray, C.T., and Eaton, M.D . (1960) J . Bacteriol . 79, 95-102 . The properties of the partially purified enzymes, including their differential behavior towards a number of protease inhibitors, are reported.

Arch Microbiol, 1979 Nov, 123(2), 137 - 41
Amino acid utilization patterns in clostridial taxonomy; Elsden SR et al.; The polyamide layer technique for the chromatographic separation of dimethylaminonaphthalene sulphonyl amino acids has been adapted to the qualitative analysis of amino acids in media before and after the growth of micro-organisms . The method has been used to study the amino acids metabolized by cultures of proteolytic clostridia growing in a medium consisting of an acid hydrolysate of casein as a source of amino acids and small amounts of yeast extract and trypticase as sources of growth factors . The chromatograms of the media after growth showed which amino acids were used and which new amino acids were produced . Clostridium botulinum type F (proteolytic), C . ghoni, C . mangenoti and C . putrificum were found to reduce proline to 5-aminovaleric acid and to produce 2-aminobutyric acid, properties they shared with C . sporogenes and C . sticklandii . C . botulinum type G and C . subterminale used glycine, lysine, serine, and arginine but in contrast to C . sticklandii they neither reduced proline to 5-aminovaleric acid nor produced 2-aminobutyric acid . Both organisms oxidized phenylalanine, tyrosine and tryptophan to phenylacetic acid, p-hydroxyphenyl acetic acid and indole acetic acid respectively . C . lituseburense and C . scatologenes used serine, threonine and arginine and produced 2-amino butyric acid and ornithine . C . lentoputrescens, C . limosum and C . malenomenatum resembled C . tetanomorphum by using glutamic acid and tyrosine . The chromatograms always showed the physiological group to which an organism belonged and in some cases were characteristic of the species.

Biochem J, 1979 Nov 1, 183(2), 471 - 4
Mechanism of formation, spectrum and reactivity of half-reduced eight-iron Clostridium pasteurianum ferredoxin in pulse-radiolysis studies and the non-co-operativity of the four-iron clusters; Butler J et al.; Reduction of fully oxidized Clostridium pasteurianum 8-Feox.,ox . ferredoxin by using pulse-radiolysis techniques yields the half-reduced species 8-Feox.,red . ferredoxin . The subsequent oxidation of 8-Feox.,red . ferredoxin with Co(NH3)5Cl2+ was studied . From a comparison with stopped-flow studies on the 2:1 Co(NH3)5Cl2+ oxidation of 8-Fered.,red . ferredoxin to the 8-Feox.,ox . form it is concluded that there is no redox co-operativity between the two 4-Fe centres in these reactions.

J Biochem (Tokyo), 1979 Nov, 86(5), 1345 - 52
Asymmetric manipulation of the membrane lipid bilayer of intact human erythrocytes with phospholipase A, C, or D induces a change in cell shape; Fujii T et al.; Changes in the membrane morphology and phospholipid content of human erythrocytes were determined after incubation of intact cells with each of various exogeneous phospholipases (PLases) . PLase A2 from Naja naja or bee venom induced crenation of the cells in parallel with hydrolysis of the membrane phosphatidylcholine (PC) . This crenated cell shape was reversed to a biconcave disc or cup-like form by a further treatment with lysophospholipase . In contrast, bacterial PLase C from Clostridium perfringens and Pseudomonas aureofaciens or fungal PLase D from Streptomyces chromofuscus induced invagination of the cells in parallel with hydrolysis of the PC . The action of the latter group of PLases on the membrane morphology was counteracted by PLase A2, and vice versa . Thus, participation of the membrane lipid bilayer in the induction of membrane conformational change and hence cell shape change was demonstrated.

J Bacteriol, 1979 Nov, 140(2), 468 - 78
Mechanism of acetate synthesis from CO2 by Clostridium acidiurici; Waber LJ et al.; Total synthesis of acetate from CO2 by Clostridium acidiurici during fermentations of hypoxanthine has been shown to involve synthesis of glycine from methylenetetrahydrofolate, CO2, and NH3 . The glycine is converted to serine by the addition of methylenetetrahydrofolate, and the resulting serine is converted to pyruvate, which is decarboxylated to form acetate . Since CO2 is converted to methylenetetrahydrofolate, both carbons of the acetate are derived from CO2 . The evidence supporting this pathway is based on (i) the demonstration that glycine decarboxylase is present in C . acidiurici, (ii) the fact that glycine is synthesized by crude extracts at a rate which is rapid enough to account for the in vivo synthesis of acetate from CO2, (iii) the fact that methylenetetrahydrofolate is an intermediate in the formation of both carbons of acetate from CO2, and (iv) the fact that the alpha carbon of glycine is the source of the carboxyl group of acetate . Evidence is presented that this synthesis of acetate does not involve carboxylation of a methyl corrinoid enzyme such as occurs in Clostridium thermoaceticum and Clostridium formicoaceticum . Thus, there are two different mechanisms for the total synthesis of acetate from CO2 by clostridia.

J Med Microbiol, 1979 Nov, 12(4), 449 - 57
Spore antigens in the classification of some clostridia; Princewill TJ; The spore antigens of Clostridium sporogenes, C . histolyticum, C . bifermentans and the butyric group were compared . By spore agglutination and fluorescent-antibody technique (FAT) the 69 strains of C . histolyticum were divided into two types: serum raised against type I (66 strains) reacted with all strains of this species but showed no cross reaction with any of the three types of C . sporogenes; serum raised against type II (three strains) did not react with strains of C . histolyticum type I but showed cross reaction with all the 66 strains of C . sporogenes type I . Thus, by antigenic analysis, spores of C . histolyticum type I were found to possess two components designated E and F; E was a type-specific component whilst F was shared by strains of type II . In addition, strains of C . histolyticum type II possessed a second component, G, which was shared by strains of C . sporogenes type I . There was no cross reaction between the precipitinogens of the spores of the two species . Five strains of C . bifermentans formed a homogeneous group as judged by spore agglutination, FAT and precipitation reactions . There was no cross reaction with any of the other proteolytic species studied . Five butyric strains also formed a homogeneous group . Two antigenic components were therefore assigned to the spore antigens of the two groups: J, for C . bifermentans and K, for the butyric strains.

Appl Environ Microbiol, 1979 Nov, 38(5), 789 - 94
Effect of sample transport systems on survival of bacteria in ground beef; Kotula AW et al.; The effects of two transport systems and cryoprotective agents on the survival of bacteria in ground beef samples were evaluated . Survival of Clostridium perfringens in ground beef samples after simulated transport (72 h) was higher (about 99%) in Dry Ice than in Trans Temp shipping units (-3 degrees C) . There were no significant differences between the two transport systems in survival of coliforms, Escherichia coli, Staphylococcus aureus, or aerobic bacteria . Mixing ground beef samples at a ratio of 1:1 (wt/vol) with 10, 20, or 30% buffered solutions of dimethyl sulfoxide or glycerol before freezing improved the survival of C . perfringens and coliforms in both transport systems . Recovery of E . coli was significantly higher with the addition of 10% dimethyl sulfoxide before Dry Ice transport . Addition of 10% dimethyl sulfoxide resulted in a 100% recovery of both S . aureus and aerobic bacteria from ground beef after simulated transport in Trans Temp shipping units . The use of cryoprotective agents can improve the survival of bacteria during transport of ground beef samples.

Arch Microbiol, 1979 Nov, 123(2), 203 - 8
NADH-dependent reduction of D-proline in Clostridium sticklandii . Reconstitution from three fractions containing NADH dehydrogenase, D-proline reductase, and a third protein factor; Schwartz AC et al.; The enzyme system from Clostridium sticklandii catalyzing the NADH-dependent reduction of D-proline was co-purified by chromatography on DEAE-cellulose at pH 8.2 and ammonium sulfate fractionation, and resolved into fractions containing three different protein components, NADH dehydrogenase, D-proline reductase and a third protein factor, by chromatography on DEAE-cellulose at pH 7.0 . Upon recombination of the fractions containing the three different protein components, the NADH-dependent reduction of D-proline was successfully reconstituted . The NADH dehydrogenase fractions oxidized NADH in the presence of artificial electron acceptors, and were inhibited by p-hydroxymercuriphenylsulfonate (50% at 80 nM) . They contained 3--4 different enzyme bands as revealed by polyacrylamide-gel electropherograms stained with the NADH-dependent reduction of 2,3,5-triphenyltetrazolium chloride . D-Proline reduction was also coupled to a leuco-methylene blue-generating system containing D-glucose and glucose-oxidase (EC 1.1.3.4) . Circumstantial evidence indicated that, among the clostridial proteins, only D-proline reductase and the third protein factor were needed for this reaction.

Vet Med (Praha), 1979 Nov, 24(11), 691 - 7
{Use of a selective sulfite-reducing medium for the isolation of Clostridium perfringens}; Konecny S et al.; The effect of sulphite-reduction in 33 sample bacterial strains was tested . With regard to the capacity of reducing sulphite in modified sulphite-reduction media in a wide scale of bacterial strains the possibility of an application of selective media with an addition of various concentrations of antibiotic solutions was checked . A concentration of 750 microgram of D-cycloserine per 1 ml of the sulphite-reduction medium appeared to be the most advantageous for the isolation and detection of sulphite-reductive clostridia, above all of Clostridium perfringens . This concentration ensured also a sufficient inhibition of undesirable bacteria without any affection of the growth and capacity of Clostridium perfringens to reduce sulphite in the applied medium.

J Med Microbiol, 1979 Nov, 12(4), 497 - 501
Catecholamine and cyclic nucleotide response of sheep to the injection of Clostridium welchii type D epsilon toxin; Worthington RW et al.; Injection of Clostridium welchii (C . perfringens) type D epsilon toxin into sheep caused large increases in catecholamine and cyclic adenosine 3',5'-monophosphate levels and moderate increases in cyclic guanosine 3',5'-monophosphate levels . Haemoconcentration also occurred . It is suggested that a rapidly developing brain oedema is the stimulus for a release of catecholamines which in turn activates adenyl cyclase . The resulting rise in cAMP causes glycogenolysis and hyperglycaemia.

J Infect Dis, 1979 Nov, 140(5), 818 - 21
Bacteriology of rattlesnake venom and implications for therapy; Goldstein EJ et al.; Although the incidence of infection secondary to the bites of venomous snakes remains unknown, the routine use of prophylactic antimicrobial therapy is advocated . In this study, the venom from 15 rattlesnakes was cultured, and 58 aerobic and 28 anaerobic strains of bacteria were isolated . The most common species isolated were Pseudomonas aeruginosa, Proteus species, coagulase-negative staphylocci, and Clostridium species . Bacteroides fragilis was also recovered . When the fang sheaths of four additional rattlesnakes were retracted and the fangs of these snakes decontaminated, 50% of the samples of venom had no bacterial growth (P = 0.035) . Until a clinical study is performed, the use of antimicrobial therapy that reflects that complex oral flora of rattlesnakes is still recommended in most cases of envenomization.

J Infect Dis, 1979 Nov, 140(5), 653 - 8
The presence of antibody-coated anaerobic bacteria in asymptomatic bacteriuria during pregnancy; Meijer-Severs GJ et al.; Quantitative anaerobic culture of urine samples obtained from 593 pregnant women by suprapubic bladder aspiration was performed to establish the involvement of anaerobic bacteria in asymptomatic urinary tract infections . The fluorescent antibody (FA) test was applied to the sediments of bladder aspirates to determine the site of infection . Anaerobic bacteriuria (greater than or equal to 10(4) microorganisms/ml of urine) was found in 34 patients, of whom five were FA-positive . These anaerobes were identified as Lactobacillus minutus, Veillonella parvula (two patients) . Clostridium putrefaciens, and Peptostreptococcus anaerobius . Aerobic bacteriuria (greater than 10(4) microorganisms/ml of urine) was detected in 27 patients, of whom 13 were FA-positive . In 10 women with mixed aerobic/anaerobic bacteriuria, no FA-positive bacteria were found . The finding of FA-positive anaerobes may indicate that these organisms are involved in silent renal infection.

Appl Environ Microbiol, 1979 Nov, 38(5), 846 - 9
Effect of processing variables on the outgrowth of Clostridium sporogenes PA 3679 spores in comminuted meat cured with sorbic acid and sodium nitrite; Robach MC; The effects of the initial pH and a "short pump" on the outgrowth of Clostridium sporogenes PA 3679 spores in comminuted cured pork were studied . Fresh ground pork was cured with salt, sugar, phosphate, ascorbate, and varying amounts of sodium nitrite and sorbic acid . The product was comminuted and inoculated with 1,000 spores of C . sporogenes per g . The meat was stuffed into 1-ounce (ca . 28.4-g) aluminum tubes, cooked to 58.5 degrees C, cooled, and incubated at 27 degrees C to observe for swells . Product cured with 0.2% sorbic acid in combination with 40 ppm sodium nitrite (40 microgram/g) had better clostridium inhibition than did product cured with 120 ppm nitrite within a pH range of 5.0 to 6.7 . The sorbic acid-40 ppm nitrite combination also gave better clostridial protection than did the 120 ppm nitrite alone when reduced amounts of curing ingredients were present.

Appl Environ Microbiol, 1979 Nov, 38(5), 767 - 71
Toxin production by Clostridium botulinum in grass; Notermans S et al.; Investigations on farms where botulism has occurred in cows showed that proteolytic Clostridium botulinum type B was present in newly made grass silages . Experiments were undertaken to study growth and toxin production of C . botulinum in grass . Of the strains tested only proteolytic strains of C . botulinum types A and B were able to produce toxin with grass as a substrate . Proteolytic strains of type B produced both medium (12S) and large (16S) toxin forms . The minimal water activity (aw) for toxin production at pH 6.5 and 5.8 was 0.94 . At pH 5.3, toxin was produced at an aw of 0.985 . These results indicate that proteolytic strains of C . botulinum (if present) may multiply and produce toxin in wilted grass silages.

J Bacteriol, 1979 Nov, 140(2), 745 - 7
Properties and function of the proton-translocating adenosine triphosphatase of Clostridium perfringens; Hasan SM et al.; Growth of Clostridium perfringens was inhibited by compounds which dissipate or prevent the formation of electrochemical proton gradients . Membrane vesicles prepared from this organism exhibited Mg2+-dependent adenosine triphosphatase (ATPase) activity sensitive to N,N'-dicyclohexylcarbodiimide . Mg2+-ATPase activity was optimal of 50 degrees C, but no discrete pH optimum was observed . Adenosine triphosphate-dependent quenching of the fluorescence of the weak base quinacrine by everted membrane vesicles suggested that the Mg2+-ATPase is a proton pump capable of generating an electrochemical proton gradient . Adenosine triphosphate-dependent transport of Ca2+ by everted vesicles was sensitive to uncouplers and inhibitors of the Mg2+-ATPase.

Biochemistry, 1979 Oct 30, 18(22), 4860 - 9
Iron-sulfur clusters in the molybdenum-iron protein component of nitrogenase . Electron paramagnetic resonance of the carbon monoxide inhibited state; Davis LC et al.; Carbon monoxide inhibits reduction of dinitrogen (N2) by purified nitrogenase from Azotobacter vinelandii and Clostridium pasteurianum in a noncompetitive manner (Kii and Kis = 1.4 X 10(-4) and 4.5 X 10(-4) and 7 X 10(-4) atm and 14 X 10(-4) atm for the two enzymes, respectively) . The onset of inhibition is within the turnover time of the enzyme, and CO does not affect the electron flux to the H2-evolving site . The kinetics of CO inhibition of N2 reduction are simple, but CO inhibition of acetylene reduction is complicated by substrate inhibition effects . When low-temperature (approximately 13 K) electron paramagnetic resonance (EPR) spectra of CO-inhibited nitrogenase are examined, it is found that low concentrations of CO ({CO} = {enzyme}) induce the appearance of a signal with g values near 2.1, 1.98, and 1.92 with t1/2 approximately 4 s, while higher concentrations of CO lead to the appearance of a signal with g values near 2.17, 2.1, and 2.05 with a similar time course . The MoFe proteins from Rhizobium japonicum and Rhodospirillum rubrum, reduced with Azotobacter Fe protein in the presence of CO, give similar results . Under conditions which promote the accumulation of H2 in the absence of CO, an additional EPR signal with g values near 2.1, 2.0, and 1.98 is observed . The use of Azotobacter nitogenase components enriched selectively with 57Fe or 95Mo, as well as the use of 13CO, permitted the assignment of the center(s) responsible for the induced signals . Only 57Fe, when present in the MoFe protein, yielded broadened EPR signals . It is suggested that the MoFe protein of nitrogenase contains one or more iron-sulfur clusters of the type found in the simple ferrodoxins . It is further proposed that the CO-induced signals arise from states of the MoFe protein in which CO inhibits electron flow to the N2-reducing site so that the iron-sulfur cluster achieves steady-state net charges of -1 (high CO complex) and -3 (low CO complex) in analogy to the normal paramagnetic states of high-potential iron-sulfur proteins and ferredoxins, respectively . The "no-CO" signal may be either an additional center or the N2-reducing site with H2 bound competitively.

Nature, 1979 Oct 4, 281(5730), 398 - 9
Clostridium botulinum can grow and form toxin at pH values lower than 4.6; Raatjes GJ et al.; It is generally accepted that in Clostridium botulinum both growth and toxin formation are completely inhibited at pH values below 4.6 . This critical pH value has been confirmed by many investigators using food as substrate or culture media . Occasionally growth of C . botulinum and toxin formation at pH values lower than 4.6 have been reported . In these cases the authors ascribed the unexpected outgrowth and toxin formation to local pH differences in inhomogeneous media and growth of C . botulinum before pH equilibration, or to the fact that fungi created microenvironments within or adjacent to the mycelial mat, where the pH was higher than 4.6 as was demonstrated by Odlaug and Pflug . We show here that the general assumption that C . botulinum does not grow below pH 4.6 is incorrect . We have observed that growth and toxin formation by C . botulinum can take place in homogeneous protein rich substrates (containing 3% or more soya or milk protein) at pH values lower than 4.6.

J Antibiot (Tokyo), 1979 Oct, 32(10), 985 - 94
Studies on bacterial cell wall inhibitors . VII . Azureomycins A and B, new antibiotics produced by Pseudonocardia azurea nov . sp . Taxonomy of the producing organism, isolation, characterization and biological properties; Omura S et al.; Two new basic water-soluble antibiotics, azureomycins A and B, were isolated from the culture broth of an actinomycete, strain AM-3696, designated as Pseudonocardia azurea nov . sp . The antibiotics exhibit moderate antimicrobial activities against Gram-positive bacteria including penicillin-resistant Staphylococcus, Mycobacterium and Clostridium . They inhibit the synthesis of bacterial cell wall peptidoglycan.

Br J Surg, 1979 Oct, 66(10), 738 - 42
Antibiotic-associated colitis--a review of 66 cases; Mogg GA et al.; We have reviewed 66 cases of antibiotic-associated colitis since March 1975, which have been associated with a 27 per cent mortality . We believe antibiotics may predispose patients to this condition which is caused by a toxin produced by Clostridium difficile . Although the disease is rare, it is more common than previously reported . The presentation, methods of diagnosis and treatment are discussed.

J Hyg (Lond), 1979 Oct, 83(2), 231 - 6
Food poisoning in hospitals in Scotland; Sharp JC et al.; A review of 50 hospital-based outbreaks of food poisoning which were reported in Scotland during 1973--7, is described . At least 1530 persons consuming hospital-prepared food were involved . Thirty-one episodes were associated with Clostridium perfringens (C . welchii), 11 were due to food-borne salmonella infection, three to enterotoxigenic Staphylococcus aureus, and five incidents were of undetermined aetiology . This differs noticeably from the experience in England and Wales where salmonellas appear to predominate as the main cause of hospital outbreaks . Twenty-two incidents occurred in hospitals for psychiatric or mentally subnormal patients, and ten others were located in geriatric units . Only 33 hospitals were involved in the 50 outbreaks as nine hospitals experienced two or more episodes . The role of the hospital in the occurrence of food poisoning may be over-emphasized in comparison with other catering establishments, as outbreaks are more readily recognized and laboratory facilities are usually available for investigation, but it is also believed that many episodes may not be reported . The peculiar problems of the hospital-catering service and particularly those of the older long-stay hospitals, are discussed in relation to preventive measures which would minimize the hazards of food poisoning.

Infect Immun, 1979 Oct, 26(1), 150 - 6
Use of ganglioside affinity filters to identify toxigenic strains of Clostridium botulinum types C and D; Hayes S; Clostridium botulinum neurotoxin is synthesized by toxic clones grown anaerobically on ganglioside affinity filters . The toxin binds to the filters and is detected by reaction with 125I-immunoglobulin G from type-specific antitoxin . Toxin spots from culture filtrates were similarly identified . The C . botulinum type C and D strains were selected for developing this affinity filter assay because synthesis of the C1 and D toxins is bacteriophage dependent . Toxigenic clones were distinguished from prophage-cured atoxigenic derivatives . These studies represent a first step toward the development of a general nonbiological screening procedure for identifying botulinal toxin and toxigenic cells . The affinity filter methodology should facilitate genetic analysis of the basis of C . botulinum toxicity.

Proc Natl Acad Sci U S A, 1979 Oct, 76(10), 4986 - 9
Identification of iron-sulfur centers in the iron-molybdenum proteins of nitrogenase; Kurtz DM Jr et al.; The core extrusion method has been applied to the determination of the type ({2Fe-2S}, {4Fe-4S}) and number of iron-sulfur centers in the FeMo proteins of the nitrogenases from Clostridium pasteurianum and Azotobacter vinelandii . The method involves extrusion with o-xylyl-alpha, alpha'-dithiol, ligand exchange of the extrusion products with p-CF3C6H4SH (RFSH), and identification and quantitation of the resultant {FenSn(SRF)4}2- complexes (n = 2,4) by 19F NMR spectroscopy . In hexamethylphosphoramide/water, 4:1 (vol/vol), 49-56% of the Fe content was extruded as {Fe4S4(SRF)4}2-, corresponding to 3,4-4.0 Fe4S4 cores per alpha 2 beta 2 subunit complex . The extruded iron does not arise from the FeMo cofactor, separate examination of which detected no extrusion products, and corresponds to 90-103% of noncofactor iron . No significant quantity of Fe2S2 cores was extruded . These results indicate the presence of four {4Fe-4S} centers per alpha 2 beta 2 subunit complex in preparations undepleted in iron . There are two main structural populations of iron atoms in these proteins, those in the cubane-type Fe4S4 cores and those in the FeMo cofactor.

Avian Dis, 1979 Oct-Dec, 23(4), 1072 - 4
The prevention of experimentally induced necrotic enteritis in chickens by avoparcin; Prescott JF; Four groups of about seventy 2-week-old broiler chickens were challenged with a Clostridium perfringens Type A isolate . Inclusion of avoparcin at 20 ppm in feed prevented necrotic enteritis in one group, but 10 ppm was only marginally effective . Bacitracin at 110 ppm also prevented the disease . Necrotic enteritis was successfully reproduced in untreated control birds.

J Bacteriol, 1979 Oct, 140(1), 59 - 64
Spore lytic enzyme released from Clostridium perfringens spores during germination; Ando Y; The exudate of fully germinated spores of Clostridium perfringens was found to contain a large amount of a spore lytic enzyme which acted directly on alkali-treated spores of the organism to cause germination . Although no detectable amount of the enzyme was found in dormant spores during germination in a KCl medium, the enzyme was produced rapidly and released into the medium . The optimal conditions for enzyme activity were pH 6.0 and 45 degrees C . Maximum activity occurred in the presence of various univalent cations at a concentration of 50 mM . The enzyme was readily inactivated by several sulfhydryl reagents . A strong reducing condition was generated in the ionic germination of the spores, a minimum Eh level of -350 mV being reached 30 min after initiation of germination . Furthermore, adenosine triphosphate-dependent pyruvate:ferredoxin oxidoreductase (EC 1.2.7.1) was identified in both dorman and germinated spores . The relationship between the release of active enzyme and the generation of reducing conditions during germination is discussed.

Zentralbl Bakteriol {Orig A}, 1979 Oct, 245(1-2), 114 - 22
A proposed sero-grouping scheme for epidemiological investigation of food poisoning due to Clostridium perfringens type A; Chakrabarty AK et al.; Serological studies with soluble and particulate antigens of Cl . perfringens type A revealed that enterotoxin and spore antigens could be used as a suitable marker for epidemiological studies . 94% of the food poisoning strains of Cl . perfringens type A could be grouped into 3 groups with the help of 2 enterotoxin-specific sera and 90% in 4 groups with antispore sera . Heat-sensitive strains were found to be antigenically more homogenous than the heat-resistant ones . Sera raised against spores of heat-resistant strains could not agglutinate spore of any of the heat-sensitive strains . Similarly no spores of heat-resistant strains were agglutinable by serum raised against spores of heat-sensitive strains . On the basis of typing efficiency the two antigen, viz enterotoxin and spore antigens were used in the serogrouping scheme proposed for the epidemiological investigation of food poisoning with Cl . perfringens type A . Used together, enterotoxin and spore agglutinogen form an antigenic formula for each strain showing the serogroup to which it belongs.

J Gen Microbiol, 1979 Oct, 114(2), 349 - 54
Characterization of the autolytic enzymes of Clostridium perfringens; Williamson R et al.; Clostridium perfringens and isolated walls of this organism autolysed rapidly when incubated in buffer at pH 7.0 with the release of free-reducing groups but no N-terminal amino acids . The predominant autolytic enzyme was an endo-beta-N-acetylglucosaminidase, and an endo-beta-N-acetylmuramidase was also present . The autolytic enzymes could be solubilized by extraction of the organisms with 5 M-LiCl and would then subsequently bind to and rapidly lyse walls of Micrococcus luteus and, more slowly, formamide-extracted walls of C . perfringens and walls of Bacillus subtilis . Lysis of C . perfringens walls by these extracted enzymes could not be demonstrated.

Appl Environ Microbiol, 1979 Oct, 38(4), 637 - 41
Effect of environmental stress on Clostridium difficile toxin levels during continuous cultivation; Onderdonk AB et al.; A method for the continuous culture of Clostridium difficile has been described . It has been shown that subjecting continuous cultures of this microorganism to environmental stress results in increased levels of toxin in culture medium . Factors found to cause this release include alteration of the Eh from --360 to +100 mV or increasing the temperature from 37 to 45 degrees C . The increased toxin levels were not associated with a change in viable cell density or the numbers of spores present . Additional studies have shown that subinhibitory concentrations of vancomycin and penicillin, but not clindamycin, also cause an increase in toxin levels during continuous culture . The increase in supernatant toxin levels occurs concomitant with a decrease in sonicated cell extract toxin levels . The data suggest that a number of factors can cause a release of toxin from C . difficile into the surrounding medium.

Appl Environ Microbiol, 1979 Oct, 38(4), 606 - 11
Toxin production by Clostridium botulinum type A under various fermentation conditions; Siegel LS et al.; The time of appearance and the quantity of toxin produced by the Hall strain of Clostridium botulinum type A were examined under various conditions . A 70-liter fermentor and a complex medium consisting of 2% casein hydrolysate and 1% yeast extract plus an appropriate concentration of glucose were employed . Optimal conditions for toxin production were as follows: a nitrogen overlay at a rate of 5 liters/min, an agitation rate of 50 rpm, a temperature of 35 degrees C, and an initial glucose concentration of 1.0% with the pH uncontrolled . Under these conditions, the maximum toxin concentration (6.3 x 10(5) mouse median lethal doses/ml) was attained within 24 h . Cell lysis was apparently not required to obtain maximum toxin concentrations under the fermentation conditions described.

Tijdschr Diergeneeskd, 1979 Sep 15, 104(18), 707 - 12
{Studies on the persistence of Clostridium botulinum on a cattle farm (author's transl)}; Notermans S et al.; In the winter of 1978-1979, the presence of Clostridium botulinum was studied on a cattle farm, on which botulism caused by feeding the animals contaminated brewers' grains occurred in 1977 . Cl . botulinum type B, the cause of mortality among cattle at the time, was detected in grass silage prepared in 1978 . This organism was not detectable in a grass silage pit dating from 1977 and made prior to the outbreak of botulism . Investigations showed that proteolytic types of C . botulinum having grass as the substrate may produce large amounts of toxin . The production of toxin in grass silage pits may, however, be prevented by a low pH in conjunction with low water activity . The present study showed that the faeces of cattle were also contaminated with Cl . botulinum type B . The degree of infection ran parallel with the degree of contamination of silage feeding in these cases.

J Biol Chem, 1979 Sep 10, 254(17), 8139 - 42
One-electron reduction of flavodoxin . A fast kinetic study; Faraggi M et al.; The reaction kinetics of fully oxidized flavodoxin from Clostridium MP with the hydrated electron have been investigated by the pulse radiolysis method . Four spectrally distinct processes have been observed with the ultimate formation of the singly reduced flavin form of the protein . The last two species obtained in the reaction sequence are spectrally similar, and are connected through a reaction which is first order . It is proposed that this reaction involves a protein conformational alteration.

Biochemistry, 1979 Sep 4, 18(18), 4024 - 30
Deoxyribonucleic acid binding studies on several new anthracycline antitumor antibiotics . Sequence preference and structure--activity relationships of marcellomycin and its analogues as compared to adriamycin; DuVernay VH Jr et al.; The deoxyribonucleic acid (DNA) binding characteristics of adriamycin and several new anthracycline glycosides, including marcellomycin, aclacinomycin, rudolfomycin, musettamycin, and pyrromycin, have been studied . The fluorescence spectra were determined for all six anthracyclines, and the fluorescence quenching effects caused by interactions with the natural DNAs poly(dAdT)--poly(dAdT) and poly(dGdC) were characterized . Binding parameters were determined by Scatchard analyses of results obtained by spectrofluorometric titrations of anthracyclines with DNA . Consistent with earlier structure--activity relationship studies of nucleic acid synthesis inhibitory effects, the results demonstrate a correlation between the length of the glycosidic side chain and DNA binding affinity . In addition, the sugar residue 2-deoxyfucose appears to confer greater DNA binding ability than do the sugars rednosamine and cinerulose when present in the terminal position of the glycosidic side chain, also in agreement with earlier studies . The sequence preference of anthracycline--DNA interaction has been examined by using DNAs of varying GC content, including the naturally occurring calf thymus DNA (43% GC), Clostridium perfringens DNA (28% GC), and Micrococcus luteus DNA (72% GC) and the synthetic double-stranded copolymers poly(dGdC)--poly(dGdC) and poly(dAdT)--POLY(DAdT) . The results demonstrate that although adriamycin shows an absolute requirement for GC sequences for DNA binding, marcellomycin and its analogues showed no such sequence requirement . Furthermore, an AT preference for DNA binding was demonstrated with marcellomycin and its analogues.

Eur J Biochem, 1979 Sep, 99(3), 593 - 603
Mechanistic and stereochemical studies on the glycine reductase of Clostridium sticklandii; Barnard GF et al.; Clostridial glycine reductase multienzyme complex which catalyses the reaction: Glycine + ADP + Pi + 2H leads to Acetate + ATP + NH3 was solubilised and fractionated essentially according to the method of Stadtman {T.C . Stadtman (1970) Methods Enzymol . 17A, 956--966} into two components: protein A and 'glycine reductase' fraction . A reconstituted system obtained by combining the two components in the presence of dithiothreitol catalysed the conversion of glycine into acetate concomitant with the phosphorylation of ADP to ATP . Using the reconstituted system, in which the unwanted enzyme activity catalyzing an exchange of the alpha hydrogen atoms of glycine with the protons of the medium had been greatly reduced, it was found that the conversion of (2RS)-{2-14C, 2-3H1}glycine (3H/14C = 7.16) into acetate (3H/14C = 7.03) was attended by the retention of both the C-2 hydrogen atoms of glycine . Conversion of (2S)-{2-2H1, 2-3H1}glycine and (2R)-{2-2H1, 2-3H1}glycine by the reconstituted system gave (2S)-acetate and (2R)-acetate respectively showing that the reductive deamination of glycine occurs through an inversion of configuration . The cumulative information available on the glycine reductase reaction is embodied in a hypothetical mechanism of action for the enzyme.

Can J Microbiol, 1979 Sep, 25(9), 987 - 90
A modified tube method for the cultivation and enumeration of anaerobic bacteria; Ogg JE et al.; A new type of tube (the Lee tube) has been developed for use in the cultivation and enumeration of obligate anaerobes . The Lee tube is a double-walled, screw-capped tube which allows the formation of a thin cylinder of agar medium between the two walls . Anaerobiosis is achieved through deoxygenation of the deep cylinder of agar during sterilization, a minimum of head space, and use of a reducing agent to absorb oxygen introduced during the inoculation procedure . For several species of Clostridium, Bacteroides fragilis, Fusobacterium necrophorum, Veillonella alcalescens, and Pectinatus cerevisiiphilus, colony counts of cultures in the Lee tubes were comparable with those obtained in pour plates incubated in a BBL GasPak system and in anaerobic roll tubes.

Onderstepoort J Vet Res, 1979 Sep, 46(3), 121 - 4
Isolation and characterization of antibodies to Clostridium perfringens epsilon toxin from hyperimmune horse serum; Worthington RW et al.; Antibodies against epsilon toxin were isolated from hyperimmune horse serum by affinity chromatography . Purified epsilon prototoxin covalently bound to Affigel 202 was used as immunosorbent, and antibodies were eluted with 6.0 M guanidine chloride . In a single run 80 mg of antibody could be recovered from a 20 microliter column of immunosorbent . The antibody was shown to belong to the IgG(T) class of immunoglobulins.

Zentralbl Bakteriol {Orig A}, 1979 Sep, 244(4), 535 - 40
The effect of nalidixic acid on microaerophilic and anaerobic bacteria . Special study of clostridia; Cancet B; Study of the sensitivity of 141 strains of anaerobic and microaerophilic bacteria to nalidixic acid shows that few bacteria are inhibited by low concentrations (Veillonella, Eikenella, most of the Clostridia) . Nalidixic acid appears to be bactericidal with respect to Clostridium perfringens, and its point of attack in the DNA is probably different from that of metronidazole.

Rev Fr Transfus Immunohematol, 1979 Sep, 22(4), 375 - 85
{Association of acquired polyagglutinabilities of types T and B . An observation}; Janot C et al.; Although T and acquired B polyagglutinabilities are not exceptional, simultaneous occurence of the two types is a rarer phenomenon . Ten days after an open heart surgery operation, the patient, age 55, had an infectious syndrom : two strains, Clostridium perfringens and Peptococcus variabilis were isolated . Simultaneously, her red cells were found to be polyagglutinable . The association of T and acquired B polyagglutinabilities could be demonstrated by serological studies of the patient's red cells, and by in vitro transformation of normal red cells using culture supernatants obtained from the two isolated strains.

J Assoc Off Anal Chem, 1979 Sep, 62(5), 1007 - 10
Comparison of Stomacher and Waring Blendor for homogenizing foods to be examined for Clostridium perfringens; Harmon SM et al.; The Colworth Stomacher Model 400 homogenizer was compared with the Waring Blendor for preparing food homogenates to be examined for Clostridium perfringens . Forty-eight samples representing 6 different food types were inoculated with C . perfringens and examined by the AOAC official first action method for enumeration of C . perfringens in foods . Identical paired specimens of each food type were blended with the 2 devices, and plate counts were made as specified in the official first action method . The effects of frozen storage on plate counts were determined by examining 24 food samples that had been stored for 3 days at -68 degrees C and homogenized both devices . Results of a statistical analysis of the experimental data indicated no significant difference overall (P greater than 0.05) in the plate counts of homogenates prepared with the Waring Blendor or the Stomacher 400, either before or after frozen storage of the food samples . However, the overall plate count average of the 48 samples was slightly higher with the Waring Blendor than with the Stomacher 400 homogenizer.

J Biol Chem, 1979 Aug 25, 254(16), 7845 - 54
Action of Arthrobacter ureafaciens sialidase on sialoglycolipid substrates . Mode of action and highly specific recognition of the oligosaccharide moiety of ganglioside GM1; Saito M et al.; A new bacterial sialidase (N-acetylneuraminate glycohydrolase, EC 3.2.1.18) isolated from the culture filtrate of Arthrobacter ureafaciens was characterized in detail with respect to its action on sialoglycolipids . Strong electrolytes had a reversible inhibitory effect on the action of the enzyme on brain gangliosides in accordance with Debye-Huckel effect of ionic environment on ionic activity, and resulted in an acidic shift and a broadening of the pH optimum . Both ionic and non-ionic detergents markedly enhanced the enzymic activity on the gangliosides, and caused an acidic shift on the pH optimum of this enzyme . Sulfhydryl groups seemed to be involved in its active site . This enzyme had a highly specific action on sialidase-resistant ganglioside GM1, showing about 100-fold higher activity on GM1 than Clostridium perfringens sialidase, the only sialidase so far reported to cleave the lipid substrate in the presence of bile salts . In the absence of detergents, the activity of A . ureafaciens sialidase on GM1 was very low . Ganglioside GM1 in either the monomeric or micelar form was hydrolyzed to asialo-GM1 by A . ureafaciens sialidase most efficiently in the presence of sodium cholate of about three times the GM1 molar concentration . The presence of detergents increased both the Km and Vmax values for ganglioside GM1 . The oligosaccharide prepared from GM1 by ozonolysis was cleaved well by this sialidase in the absence of detergents, and no detergent was found to affect the hydrolysis . The Km value for the sugar substrate was about two orders of magnitude greater than that for the corresponding lipid substrate . It is suggested that the hydrophobic ceramide moiety increases affinity of the lipid substrate to the enzyme, but inhibits hydrolysis of the substrate, possibly due to its hydrophobic interaction with hydrophobic portions of the enzyme molecule (resulting in lower Km and Vmax for lipid substrates) . This inhibition may be released by detergent due to formation of mixed micelles of sialoglycolipid and detergent molecules . It is also indicated that recognition of the specific saccharide structure of GM1 by individual sialidases is essential for release of the resistant sialyl residue, and that A . ureafaciens sialidase seemed to have an isoenzymic or oligomeric structure.

Biochim Biophys Acta, 1979 Aug 14, 547(2), 411 - 6
Chelating agents protect hydrogenase against oxygen inactivation; Klibanov AM et al.; The effect of chelation on rate or air inactivation of hydrogenase from Clostridium pasteurianum has been investigated . All chelating agents used, whether water-soluble or water-insoluble, afforded protection against oxygen inactivation . EDTA appeared to be the most effective . Thus, in the absence of EDTA, hydrogenase in aqueous solution was nearly totally inactivated after 1 hour incubation in air, whereas 0.5 M EDTA (which did not affect significantly catalytic activity) allowed 41% retention of the initial activity even after 3 days incubation.

Z Exp Chir, 1979 Aug, 12(4), 209 - 15
{Tumor localization in vivo on the basis of scintigraphic detection of clostridium rods by means of 131J-labeled antibodies and F(ab')2 antibody fragments}; Vogt R et al.; The authors report about investigations for localization of tumours in vivo by scintigrafic iddntification of Clostridium butyricum Jena H8 by 131I-labelled antibodies and by 131I-labelled F(ab')2-fragment antibodies in mouse transplantation-tumour UVT 15264 . The application of 131I-labelled anti-clostridial-rod-antibodies in tumour bearing mice demonstrates increase of radioactivity in tumours of animals, pretreated by spores, as well as in animals without pretreatment . These appearently unspecific increases of radioactivity could not be demonstrated by application of 131I-labelled F(ab')2-fragment antibodies directed against clostridial rods, but also the specific fixation of 131I-labelled F(ab')2-fragment antibodies in tumours of mice, pretreated by clostridial spores, could not be demonstrated . The results are discussed with regard to further investigations.

Appl Environ Microbiol, 1979 Aug, 38(2), 197 - 9
Action of egg white lysozyme on Clostridium tyrobutyricum; Wasserfall F et al.; A 500-U ml-1 portion of egg white lysozyme was able to kill 99% of 5 X 10(5) resting vegetative cells of Clostridium tyrobutyricum within 24 h of incubation at 25 degrees C . Spores were completely resistant to lysozyme . Proliferating vegetative cells were severely inhibited, although lysozyme-resistant cells developed in growing cultures in the presence of lysozyme . Whereas early stages of spore germination (loss of optical refractility and heat resistance) were not inhibited by lysozyme, the overall outgrowth of spore cells into vegetative cells was delayed by 1 day in the presence of 500 U of lysosyme ml-1 . This delay was independent of the lysozyme sensitivity or resistance of the mother culture of the used spores . It is suggested that this inhibition by lysozyme of the outgrowth of spore cells into vegetative cells of the lactate-fermenting C . tyrobutyricum is the basis for the observation that lysozyme can substitute for nitrate in preventing the "late gas" defect of Edam- and Gouda-type cheeses.

J Clin Microbiol, 1979 Aug, 10(2), 188 - 91
Obligate anaerobes in clinical veterinary practice; Hirsh DC et al.; Clinical specimens obtained from domestic animals were examined to determine the relative prevalence of obligate anaerobic bacteria and the species represented . Of 3,167 samples cultured anaerobically as well as aerobically, 2,234 were bacteriologically positive . Of these positive samples, 583 (26%) contained species of obligate anaerobic bacteria in a total of 641 isolates . Most positive samples contained anaerobes admixed with aerobic species, although 6% of such samples yielded pure cultures of obligate anaerobes . The most common sites from which anaerobes were isolated were abscesses (32% of abscesses cultured contained species of obligate anaerobes), peritoneal exudates (24%), and pleural effusions (20%) . Bacteroides melaninogenicus, Bacteroides spp., Peptostreptococcus anaerobius, and Bacteroides ruminicola accounted in the aggregate for approximately 50% of all anaerobic isolates . Bacteroides fragilis accounted for 1% of all the isolates, and members of the genus Clostridium accounted for 8%.

Can J Biochem, 1979 Aug, 57(8), 1093 - 8
The interaction of polymeric viologens with hydrogenases from Desulfovibrio desulfuricans and Clostridium pasteurianum; Glick BR et al.; The interaction between hydrogenases from either Desulfovibrio desulfuricans or Clostridium pasteurianum and electron donors methyl viologen or polymeric viologens was examined . Extracts from each organism contained a single gel electophoretic band of active hydrogenase . The hydrogenase of D . desulfuricans was much more stable than that of Cl . pasteurianum . With methyl viologen apparent Km and Vm values were 0.5 mM and 0.62 mumole H2/min per milligram protein for the Cl . pasteurianum and 0.7 and 6.2 mumole H2/min per milligram protein, respectively, for the D . desulfuricans enzyme . The hydrogenases bound the polymeric viologens more tightly than methyl viologen, more so for the enzyme of D . desulfuricans than for Cl . pasteurianum . Maximal rate of hydrogen production was less with the polymeric than with methyl viologen . The results suggest that the D . desulfuricans enzyme in conjunction wiion than that from Cl . pasteurianum.

Appl Environ Microbiol, 1979 Aug, 38(2), 297 - 300
Noncorrelation between mouse toxicity and serologically assayed toxin in Clostridium botulinum type A culture fluids; Betley MJ et al.; Toxicity in culture fluids of several Clostridium botulinum type A strains was assayed in mice and converted to weight equivalent . The toxin-related antigen in the samples was quantitated by a radioimmunoassay which used standards of known antigen concentration instead of the usually used toxicity . Freshly prepared samples had reasonably similar titers of toxin and antigen . When the samples were held at room temperature for several weeks, toxicity decreased more than antigenicity, but the relative decreases of the two varied with the samples . The results are discussed as evidence that serological assays of botulinum toxin cannot always be used for accurate determination of toxicity.

Appl Environ Microbiol, 1979 Aug, 38(2), 216 - 8
Bile acid inhibition of Clostridium botulinum; Huhtanen CM; Bile acids and their glycine and taurine conjugates were tested in vitro for inhibition of Clostridium botulinum types A and B . Cholic acid inhibited most strains at 2 mg/ml, whereas chenodeoxycholic acid inhibited all strains at 0.4 mg/ml . Deoxycholic acid inhibited one strain at 0.08 mg/ml and other strains at 0.4 and 2 mg/ml . Lithocholic acid inhibited all strains at 0.016 mg/ml . Glycine conjugates also showed considerable inhibition of some strains, whereas taurine conjugates were inactive.

J Hyg (Lond), 1979 Aug, 83(1), 1 - 9
Immunofluorescent study of the spore antigens of proteolytic strains of Clostridium botulinum; Princewill TJ; By means of the spore fluorescent antibody technique 31 strains of Clostridium botulinum types A (18 strains), B (10 strains) and F (3 strains) were found to belong to the same homogeneous group irrespective of their toxigenic types . Some strains of this species also cross-reacted with certain strains of Clostridium sporogenes types I, II and III and Clostridium histolyticum type II . By spore antigenic analysis it was found that Clostridium parabotulinum contained two components designated L and M, the former describing species specificity; the latter was the cross-reacting component shared by some strains of Clostridium sporogenes and Clostridium histolyticum . Following this, a scheme showing the distribution of spore antigenic components among various species of Clostridium was given.

Can J Microbiol, 1979 Aug, 25(8), 947 - 8
A new nitrogen-fixing Clostridium species from a high Arctic ecosystem; Jordan DC et al.; A hitherto undescribed species of yellow-pigmented, Gram-negative Clostridium sp., possessing nitrogenase activity, has been isolated from a number of sampling sites on the Truelove Lowland of Devon Island in the Canadian high Arctic . This bacterium, tentatively designated Clostridium arcticum sp . nov., accounted for 19% of all isolates recovered which were capable of anaerobic nitrogen fixation.

Aust N Z J Med, 1979 Aug, 9(4), 426 - 9
Necrotising enterocolitis associated with invasion by Clostridium septicum complicating cyclic neutropaenia; Bignold LP et al.; A fatal case of necrotising enterocolitis complicating cyclic neutropaenia in a 17-year-old boy is reported . The episode of enterocolitis was characterised by fulminant, generalised peritonitis associated with necrosis and gas formation in the wall of the ileum and caecum . Clostridium septicum was grown from premortem blood cultures and Gram positive bacilli typical of this organism were present in histological sections of the bowel wall . Necrotising enterocolitis in this and previously reported cases of cyclic neutropaenia resembles the agranulocytic form of intestinal necrosis which occasionally complicates leukaemia.

Biochem J, 1979 Aug 1, 181(2), 387 - 99
Effect of O-sulphate groups in lactose and N-acetylneuraminyl-lactose on their enzymic hydrolysis; Mian N et al.; 1 . Lactose 6'-O-sulphate, N-acetylneuraminyl-(alpha 2 leads to 3)-D-lactose 6'-O-sulphate, N-acetylneuraminyl ?-O-sulphate-(alpha 2 leads to 3)-D-lactose 6'0-O-sulphate, N-acetylneuraminyl ?-O-sulphate-(alpha 2 leads to 6)-D-lactose and N-acetylneuraminyl-(alpha 2 leads to 3)- and -(alpha 2 leads to 6))-lactose 6'-O-sulphate were prepared by chemical sulphation of lactose, N-acetylneuraminyl-lactose and tis isomers by using pyridine-SO3 reagent . 2 . Significant kinetic differences were observed in the enzymic hydrolysis of the sulphated derivatives compared with unsubstituted substrates . 3 . In the case of reactions catalysed by rat liver lysosomal and Clostridium perfringens neuraminidases (EC 3.2.1.18), the presence of an O-sulphate group in the N-acetylneuraminyl moiety affected the reaction by decreasing the Km and the Vmax, its presence in the galactosyl moiety affected the reaction by decreasing the Km and increasing the Vmax . and its presence in both N-acetylneuraminyl and galactosyl moieties decreased the Km and the Vmax . of the reaction . 4 . Mixed-substrate reaction kinetic data indicated competition between the sulphated and unsubstituted substrates for the same active sites on the neuraminidase molecule . 5 . Lactose 6'-O-sulphate neither behaved as a substrate nor acted as an inhibitor with respect to unsubstituted lactose and p-nitrophenyl beta-D-galactopyranoside when tested with lactase of suckling rat intestine and Escherichia coli beta-D-galactosidase (EC 3.2.1.23) . 6 . Preliminary investigation also indicated that, whereas glucose 6-O-sulphate and glucose 3-O-sulphate were were neither substrate nor inhibitor of glucose oxidase (EC 1.1.3.4), galactose 6-O-sulphate was oxidized half as fast as unsubstituted galactose by galactose dehydrogenase (EC 1.1.1.48).

Biochem J, 1979 Aug 1, 181(2), 377 - 85
Neuraminidase inhibition by chemically sulphated glycopeptides; Mian N et al.; Chemically sulphated glycopeptides (derived from pig duodenal mucosa) inhibited Clostridium perfringens neuraminidase (EC 3.2.1.18) activity in a pH-dependent manner . Analysis of inhibition kinetics data indicated that, although the enzyme inhibition could not be categorized into any of the classical types of inhibition, it could be interpreted as a function of the size and shape of the substrates used . The enzyme activity was inhibited by 86% and 40% when tested with bovine submaxillary-gland mucin (mol . wt . 4 x 10(5)-40 x 10(5) and N-acetylneuraminyl-lactose (mol . wt . 633) as substrates respectively . Presence of sulphated glycopeptide did not affect the binding of N-acetylneuraminic acid (mol . wt . 309), a competitive inhibitor of Vibrio cholerae neuraminidase, to the enzyme active site . The enzyme inhibition was thus considered to be due to steric hindrance as a consequence of the non-specific interactions between the enzyme molecule and polyanionic sulphated glycopeptide affecting the differential accessibility of the substrate molecules to the enzyme active site . The enzyme-inhibitor interaction could be suppressed by rapid and many-fold dilution of the reaction mixture, by concurrent addition of the inactive enzyme or by partial removal of the sulphate esters from the sulphated glycopeptide molecule by the action of Helix pomatia arylsulphatase (EC 3.1.6.1).

Helv Chir Acta, 1979 Aug, 46(3), 477 - 81
{Acute emphysematous cholecystitis}; Huber T et al.; Emphysematous cholecystitis is a rare form of acute cholecystitis, characterized radiographically by the presence of gas within the gallbladder . We report of a patient, who was admitted to the hospital with the diagnosis of acute abdomen . This patient had an emphysematous cholecystitis caused by Clostridium perfringens . We found the wall of the gallbladder emphysematous and gangrenous, the gallbladder was distended and contained purulent material, but no stones . However, in addition, the films of abdomen showed gas in the ducts . Diagnosis, pathogenesis and the aetiological and therapeutical aspects will be discussed.

Jpn J Med Sci Biol, 1979 Aug, 32(4), 199 - 205
Enzyme linked immunosorbent assay (ELISA) for detection of Clostridium botulinum type B toxin; Kozaki S et al.; The enzyme-linked immunosorbent assay using different techniques has been applied to determine botulinum type B toxin . With the so-called "sandwich" technique, about 5,000 mouse ip LD50 of type B toxin can be detected . With the "double-sandwich" technique, about 400 mouse ip LD50 of toxin is detected and different commerical antisera are useful . For accurate quantification of botulinum toxins in culture filtrates, addition of EDTA to samples seems to be necessary . Cross-reactivity of the assay depends on the specificity of the antisera against botulinum type B toxin used and is almost eliminated with antiserum prepared against the toxic component of type B toxin.

Eur J Biochem, 1979 Aug 1, 98(2), 597 - 612
The proton-translocating adenosine triphosphatase of the obligately anaerobic bacterium Clostridium pasteurianum . 1 . ATP phosphohydrolase activity; Clarke DJ et al.; 1 . The cell-membrane ATP phosphohydrolase of vegetatively grown Clostridium pasteurianum was specifically Mg2+-dependent, but demonstrated significant activity with GTP, CTP and UTP . It displayed approximate Michaelis-Menten kinetics only in the presence of certain effectors (e.g . phosphoenolpyruvate, fructose 1,6-bis-phosphate) which decreased the Km for ATP (to below 2 mM) but also V, whilst extending to pH 5.8 the effective pH range of activity of the enzyme . 2 . ATP phosphohydrolase activity of the membrane ATPase (BF0F1) was inhibited by N,N'-dicyclohexylcarbodiimide, butyricin 7423, Dio-9, 4-chloro-7-nitrobenzofurazan, efrapeptin, leucinostatin and quercetin, and to a lesser degree by aurovertin and citreoviridin . The enzyme was not inhibited by oligomycin, spegazzinine, tributyl tin, triethyl tin or venturicidin . The soluble ATPase (BF1) component differed in not being inhibited by N,N'-dicyclohexylcarbodiimide, butyricin 7423 or leucinostatin . 3 . The ATPase (BF0F1) complex and its soluble (BF1) component were separately purified . 4 . Dodecylsulphate/polyacrylamide gel electrophoresis separated only four polypeptide components in the purified ATPase (BF0F1), with approximate molecular weights (+/- 10%) as follows: subunit a, 65 500; subunit c, 57 500; subunit da, 43 000; subunit fa, 15 000 . The soluble (BF1 component contained only the three polypeptide subunits a, c and da . These were present in the BF0F1 preparation in the ratio 2 : 1 : 2; the contribution of subunit fa could not satisfactorily be quantified . 5 . Subunit a was identified as the component binding 4-chloro-7-nitrobenzofurazan and subunit fa as the component binding N,N'-dicyclohexylcarbodiimide . The ATP phosphohydrolase activity of the membrane ATPase was not activated by trypsin treatment and the ATPase (BF0F1) contained no trypsin-sensitive inhibitor protein subunit . 6 . Purified ATPase (BF0F1) was incorporated into artificial proteoliposomes which demonstrated ATP-dependent enhancement of 8-anilinonaphthalene-1-sulphonate fluorescence and ATP-dependent proton influx . These reactions were abolished by proton conductors (e.g . carbonylcyanide m-chlorophenylhydrazone) by valinomycin in the presence of a high external concentration of K+, or by N,N'-dicyclohexylcarbodiimide, butyricin 7423, Dio-9, 4-chloro-7-nitrobenzofurazan or leucinostatin . Oligomycin, tributyl tin, triethyl tin and venturicidin were not inhibitory . 7 . When stripped of the soluble BF1 component, such ATPase-proteoliposomes demonstrated nil ATP phosphohydrolase activity and did not display ATP-dependent enhancement of 8-anilino-naphthalene-1-sulphonate fluorescence or ATP-dependent protein influx . All of these activities were restored by incubation of the BF1-depleted proteoliposomes with a purified preparation of the soluble BF1 component.

Naunyn Schmiedebergs Arch Pharmacol, 1979 Jul 31, 308(1), 67 - 70
Interaction of botulinum type A, B and E derivative toxins with synaptosomes of rat brain; Kozaki S; Clostridium botulimum 125I-labelled derivative toxin immediately bound to rat synaptosomes . Of the two fragments of type B derivative toxin, the large-molecular-weight fragment (fragment I) inhibited the binding of labelled type B derivative toxin to synaptosomes in the same manner as unlabelled type B toxin did . The inhibition by the small-molecular-weight fragment (fragment II) was less than that by fragment I . These findings suggest that type B toxin binds to synaptosomes mainly with some part of fragment I . The binding of labelled type A and E derivative toxins was inhibited by either of the unlabelled type A or E derivative toxins, but not by type B derivative toxin . It is concluded that synaptosomes of rat brain possess relatively specific binding sites for botulinum toxin types.

Mol Cell Biochem, 1979 Jul 31, 26(2), 111 - 22
Oxidative inactivation of the molybdenum-iron-protein component of nitrogenase from clostridium pasteurianum; Gomez-Moreno C et al.; The sensitivity of the molybdenum-iron(MoFe)-protein of Clostridium pasteurianum nitrogenase toward oxidation has been studied by determining the enzymatic activity of this component after incubating it anaerobically in ferricyanide solutions of various oxidizing strengths (as measured by their oxidation potentials) . It was found that the MoFe-protein remains active at potentials up to +350 mV (vs . standard hydrogen electrode) but becomes readily inactivated at more oxidizing potentials, after a lag period, depending on the potential level and temperature . Oxidative inactivation by ferricyanide results in the release of most of the Mo, Fe and S atoms from the protein which causes the loss of the absorption bands in the visible region . The metals and sulfur could be re-incorporated by incubation in a mixture containing thiol, sulfide, molybdate, and ferric iron . The EPR spectrum of the oxidatively inactivated MoFe-protein showed that both the high- and low-field signals are readily affected . Re-incorporation of the metals and sulfur into the "bleached" protein produced an EPR spectrum similar to that of the air-inactivated protein . Incubation of the Mo-Fe-protein with mersalyl abolished its enzymic activity . The difference spectrum before and after mersalyl treatment resembles that of the soluble spinach ferredoxin.

Rev Infect Dis, 1979 Jul-Aug, 1(4), 614 - 24
The clinical spectrum of infant botulism; Arnon SS et al.; Infant botulism is the systemic illness that results when spores of Clostridium botulinum germinate in the infant's intestine and then produce botulinal toxin in vivo . As with other infectious diseases, infant botulism has a spectrum of clinical severity that ranges from a mild, outpatient illness to fulminant, sudden death . Most cases reported to date have been recognized in infants so weak and hypotonic that their need for hospital care was unquestioned; yet even this group of patients displayed a wide range in severity of illness . The outpatients were initially considered to be cases of "failure to thrive," while the fulminant cases were indistinguishable at autopsy from typical instances of the sudden infant death syndrome (SIDS, crib death) . This article discusses the observed spectrum of clinical severity, the management of the hospitalized patient, and the manner in which sudden death might result from production of butulinal toxin in the intestine.

Infect Immun, 1979 Jul, 25(1), 191 - 201
Purification and characterization of Clostridium difficile toxin; Rolfe RD et al.; Recent evidence indicates that toxigenic Clostridium difficile strains are a major cause of antimicrobial-associated ileocecitis in laboratory animals and pseudomembranous colitis in humans . C . difficile ATCC 9689 was cultivated in a synthetic medium to which 3% ultrafiltrated proteose peptone was added . Purification of the toxin from broth filtrate was accomplished through ultrafiltration (100,000 nominal-molecular-weight-limit membrane), precipitation with 75% (NH4)2SO4, and chromatographic separation using Bio-Gel A 5m followed by ion-exchange chromatography on a diethylaminoethyl-Sephadex A-25 column . The purified toxin displayed only one band on polyacrylamide gel electrophoresis, and approximately 170 pg was cytopathic for human amnion cells . The isolated toxin was neutralized by Clostridium sordelli antitoxin, heat labile (56 degrees C for 30 min), and inactivated at pH 4 and 9; it had an isoelectric point of 5.0, increased vascular permeability in rabbits, and caused ileocecitis in hamsters when injected intracecally . Treatment of the toxin with trypsin, chymotrypsin, pronase, amylase, or ethylmercurithiosalicylate caused inactivation, whereas lipase had no effect . By gel filtration, its molecular weight was estimated as 530,000 . Upon reduction and denaturation, the toxin dissociated into 185,000- and 50,000-molecular-weight components, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Extensive dissociation yielded only the 50,000-molecular-weight component . The toxin appears to be protoplasmic and is released into the surrounding environment upon autolysis of the cells . Attempts to correlate specific enzymatic activity with the toxin have been unsuccessful . These studies will help delineate the role of C . difficile toxin in antimicrobial-associated colitis and diarrhea.

Eur J Biochem, 1979 Jul, 97(2), 555 - 64
Monomer-micelle transition of the ganglioside GM1 and the hydrolysis by Clostridium perfringens neuraminidase; Rauvala H; The action of Clostridium perfringens neuraminidase on the ganglioside Gm1 tritiated in the ceramide moiety was studied . The rates of hydrolysis of the Gm1 ganglioside were determined from radioactivity in the neutral glycolipid product, which was separated from the substrate on DEAE-Sephadex columns . In order to study the physical state of the substrate in the conditions used in the neuraminidase treatment, the critical micelle concentrations of the Gm1 ganglioside were determined using formation of the triiodide anion in aqueous iodine solution as an indicator . The critical micelle concentrations were also obtained by determining the non-sedimenting radioactivity at different concentrations of the labeled ganglioside per total volume used in ultracentrifugation experiments . In addition, the concentrations of the monomeric ganglioside were concluded from the results of the ultra-centrifugation studies . The increase in the reaction rate of the Gm1 hydrolysis as the function of the substrate concentration was leveled off at 25-28 microM ganglioside . The abrupt change at this concentration is interpreted as reflecting the monomer-micelle transition of the ganglioside in the conditions used (50mM sodium acetate buffer, pH 4.6) . The critical micelle concentration was 29 microM on the basis of the triiodide test, and ultracentrifugation revealed the critical micelle concentration 28 microM . The reaction velocity of the hydrolysis was decreased immediately above the critical micelle concentration, and became constant at higher concentrations of the ganglioside . A close correlation to these changes in the reaction rate is suggested to exist in the concentrations of the monomeric Gm1 ganglioside . Saturation of the buffer used in the neuraminidase assays with butanol effected a striking change in the plot of reaction rate versus ganglioside concentration . The reaction rate increased up to 100-110 microM Gm1 ganglioside . The shift of the inflexion point in the rate plot from 25-28 microM to 100-110 microM ganglioside concentration is suggested to be due to a respective change in the critical micelle concentration effected by butanol . N-Acetylneuraminyllactosyl ceramide, lactosyl ceramide and asialo-Gm1 ganglioside had an inhibitory effect on the reaction . In contrast, N-acetylneuraminyllactose, lactose and some other free saccharides were not inhibitory . The results demonstrate that factors other than the saccharide structure must be taken into account when substrate specificity of a glycosidase is studied using competition experiments . It is suggested that the inhibition effected by the glycolipids is due to an increase in the micellar state of the Gm1 ganglioside.

Rev Infect Dis, 1979 Jul-Aug, 1(4), 693 - 7
Food and environmental aspects of infant botulism in California; Chin J et al.; In an effort to identify vehicles by which Clostridium botulinum spores might have reached the intestine of patients with infant botulism, 555 samples of foods, drugs, and environmental specimens were examined . Of the food items, C . botulinum was only found in nine of 90 (10%) honey specimens . Five patients had been exposed to honey that contained C . botulinum, and ingestion of honey was found to be a significant risk factor for type B infant botulism (P = 0.005) . In addition, C . botulinum was isolated from five samples of soil (three from case homes, two from control homes) and from vacuum cleaner dust from one case home . In every instance in which C . botulinum was isolated from a specimen of honey, soil, or duct associated with a case of infant botulism, the type of toxin (A or B) in the honey, soil, or dust isolate matched the type of toxin of the organism recovered from the infant . Isolation of C . botulinum from the soil of homes of control infants emphasizes the ubiquitous distribution of and exposure to this organism and suggests that host factors are important in the development of illness . Prevention of infant botulism will depend on the identification of these host factors, as well as on the identification of other vehicles that, like honey, may convey C . botulinum spores to susceptible infants.

Rev Infect Dis, 1979 Jul-Aug, 1(4), 683 - 8
Animal models for the study of infant botulism; Sugiyama H; Intestinal infection with Clostridium botulinum was produced by intragastric administration of C . botulinum spores in conventionally reared mice seven to 13 days old but not in younger or older mice . The 50% infective dose of one of the culture strains administered was 170 spores per nine-day-old mouse . Overt botulism did not develop in these animals, but infection with C . botulinum was evidenced by the presence of botulinal toxin in the colon for up to seven days after challenge . Infant mice were at least as sensitive to the lethal action of botulinal toxin as were adult mice, and evidence suggests that infant rats may have a similar age-related susceptibility to enteric botulinal infection . Germfree adult mice were very susceptible to infection with C . botulinum, acquiring intestinally infective doses of airborne spores . Within a few days after exposure to normal mice, the axenic mice became resistant to challenge with 10(5) C . botulinum spores.

Rev Infect Dis, 1979 Jul-Aug, 1(4), 637 - 41
Clostridium botulinum: characteristics and occurrence; Smith LD; Clostridium botulinum is not a well-defined species of bacterium . Instead, it is a conglomerate of four culturally distinct groups of organisms that, among them, produce seven serologically distinct toxins, all with similar pharmacological action . The principal habitat of C . botulinum is the soil, although its distribution in the soil is sometimes highly regional . Infant botulism is caused by two types of C . botulinum: type A and the proteolytic strains of type B . Type A strains, to whose toxin humans seem most susceptible, are found most frequently in the soil of the western United States; type B strains are somewhat more universally distributed, with a higher frequency of isolation from the soil of some Appalachian areas . The frequency of occurrence of type A and type B food-borne botulism parallels the distribution of these types in the soil.

J Gen Microbiol, 1979 Jul, 113(1), 29 - 35
Taxonomy of Clostridium tetani and related species; Nakamura S et al.; Clostridium tetani and its related species C . tetanomorphum, C . cochlearium and C . lentoputrescens were examined for DNA-DNA homology and biochemical properties . Two distinctly different groups were included under the name of C . tetanomorphum: one was identical with C . cochlearium and the name C . tetanomorphum was applied to the other group with some amendment of biochemical properties . Comparison of the type strain of C . lentoputrescens with wild strains obtained from horse faeces indicated that the name C . lentoputrescens should be abolished as a later synonym of C . cochlearium . Liquefaction of gelatin and spore shape, which have been used as the important criteria for differentiation of C . tetani-related species, were genetically insignificant.

J Clin Microbiol, 1979 Jul, 10(1), 14 - 8
API and Minitek systems in identification of clinical isolates of anaerobic gram-negative bacilli and Clostridium species; Hanson CW et al.; A comparison of the API and Minitek methods of biochemical testing was made on a variety of anaerobic bacteria . Although API and Minitek results were not compared to more standardized or conventional procedures of identification, multiple repeat testing of the two systems was done on routine clinical isolates and known organisms to determine (i) whether the reactions were reliably consistent, (ii) the ease of reading the two systems with respect to the frequency of questionable results, and (iii) the percentage of routine clinical isolates for which each system yielded an identification . The Minitek system gave a much lower incidence of difficult to interpret reactions . The two systems were comparable in terms of reproducibility and capability of yielding an identification of the anaerobic gram-negative bacilli and Clostridium species, but were unsatisfactory for routine use on most of the other anaerobic bacteria isolated.

Biochem J, 1979 Jun 15, 180(3), 647 - 54
Unaltered catabolism of desialylated low-density lipoprotein in the pig and in cultured rat hepatocytes; Attie AD et al.; Removal of the terminal sialic acid residues from many serum glycoproteins results in exposure of their penultimate galactose residues and rapid clearance from circulation by the liver . Low-density lipoprotein is a glycoprotein containing 21 galactose and 9 sialic acid residues per particle . Studies in this laboratory and others have shown that both the liver and extrahepatic tissues contribute to the degradation of low-density lipoprotein . This study was undertaken to determine whether desialylation of pig low-density lipoprotein alters its removal from circulation . Low-density lipoprotein was incubated at 37 degrees C with an agarose-bound neuraminidase, proteinase-free, from Clostridium perfringens . After 18 h at pH 5.0, 70% of the sialic acid residues were removed . The desialylated 131I-labelled and native 125I-labelled low-density lipoproteins were simultaneously injected into a pig, and their disappearance from plasma was followed for 96 h . The turnovers of the two were identical . In contrast, neuraminidase-treated fetuin was cleared about 200-fold faster than native fetuin . Studies were also performed in cultured rat hepatocytes . Rates of degradation of native and neuraminidase-treated low-density lipoprotein were similar, whereas asialo-fetuin was degraded at six to ten times the rate of native fetuin . Thus desialylation does not appear to alter low-density-lipoprotein catabolism by hepatic or extrahepatic cells.

Fortschr Med, 1979 Jun 14, 97(22), 1040 - 3
{Intestinal gas gangrene originating from the gallbladder: diagnosis, therapy, report of 2 cases}; von Gottberg C et al.; The problems of diagnosis and therapy of gas gangrene of the gastrointestinal tube and the gallbladder are discussed . 2 cases of gas edema of the gallbladder due to clostridium perfringens are reported . One of the patients survived . The early diagnosis of gas gangrene is important because of the extraordinary way of infection and the necessary early surgical and medicamentous therapy.

Biochim Biophys Acta, 1979 Jun 13, 554(1), 68 - 75
Alteration of human erythrocyte plasma membranes by perfringolysin O as revealed by freeze-fracture electron microscopy . Studies on Clostridium perfringens exotoxins V; Mitsui K et al.; When human erythrocyte membranes were treated with perfringolysin O (Clostridium perfringens theta-toxin) and examined by electron microscopy after freeze-fracture, two ultrastructural alterations were observed in fracture faces of membrane . (1) A random aggregation of intramembranous particles was seen in the fracture face of the protoplasmic half (PF face) of all membranes treated with the toxin, even if at a low concentration (40 hemolytic units/ml) . On the other hand, the aggregation in the fracture face of the exoplasmic half (EF face) was observed only in membranes treated with a high concentration (3300 hemolytic units/ml) for 2 h . (2) Round protrusions and "cavities" with 30 nm in diameter were visible in EF and PF faces of membranes treated with a high concentration, respectively . These structures were always protruded toward cytoplasmic side, but did not appear to form holes through the membrane . Ring and arc shaped structures with a dark center of 26 nm and a distinct border of 5 nm in width were observed when the toxin alone was negatively stained at a very high concentration (170,000 hemolytic units/ml) . These structures were also produced in the presence of cholesterol even if the toxin concentration was low.

Biochim Biophys Acta, 1979 Jun 13, 554(1), 125 - 32
Accessibility of sialo components in a murine tumor cell to extracellular N-acetylneuraminate glycohydrolase (sialidase); Barton NW et al.; Lipid-bound sialic acid in the murine melanoma cell is not totally inaccessible to an exogenous macromolecular probe, as formerly believed . Roughly 30% of the dialic acid bound to lipid, and an equal proportion of the sialic acid bound to protein is cleaved by the action of Clostridium perfringens N-acetylneuraminate glycohydrolase (neuraminidase, sialidase) when the purified enzyme is added to the suspenion medium of intact murine melanoma cells freshly derived from the tumor . Cleavage of lipid-bound sialic acid is indifferent to the presence of Ca (2+) in the medium . However, maximum release from protein requires a physiological concentration of this divalent cation . Variation in ionic strength has no effect on release of sialic acid . These findings show that restricted portion of the bound sialic acid may be released from the intact murine melanama cell by the extracellularly supplied enzyme acting topographically.

J Biol Chem, 1979 Jun 10, 254(11), 4499 - 501
Determination of cooperative interaction between clusters in Clostridium pasteurianum 2 (4Fe-4S) ferredoxin; Sweeney WV et al.; The effect of reducing one 4Fe-4S cluster in Clostridium pasteurianum 2 (4Fe-4S) ferredoxin on the reduction potential of the unreduced cluster has been investigated . While such an effect is suggested by both the x-ray structure of Peptococcus aerogenes 2 (4F-4S) ferredoxin and the polypeptide conformational change on reduction present in clostridial-type 2 (4Fe-4S) ferredoxins, present studies indicate that cluster-cluster cooperative interaction is not strong enough to be of functional importance in these proteins.

S Afr Med J, 1979 Jun 9, 55(24), 986 - 8
Postoperative Clostridium septicum dermatitis simulating toxic epidermal necrolysis; Findlay GH et al.; Widespread detachment of the epidermis as a result of Clostridium septicum infection after a laparotomy for intestinal obstruction is described . A clinicopathological picture, hitherto undescribed, is outlined.

Arch Microbiol, 1979 Jun, 121(3), 255 - 60
Selenium requirement for active xanthine dehydrogenase from Clostridium acidiurici and Clostridium cylindrosporum; Wagner R et al.; The xanthine dehydrogenase of Clostridium acidiurici and C . cylindrosporum was assayed with methyl viologen as acceptor . In C . acidiurici the basal activity level was about 0.3 mumol/min x mg of protein . Cells grown on uric acid in the presence of 10(-7) M selenite showed a 14-fold increase in xanthine dehydrogenase activity, which decreased with higher selenite concentrations (10(-5) M) . The supplementation with 10(-7) M molybdate or tungstate was without effect . High concentrations of tungstate decreased the xanthine dehydrogenase if selenite was also present . In comparison, high concentrations of molybdate affected only a small decrease in activity level at the optimal concentration for selenite and relieved to some degree the inhibitory effect of 10(-5) M selenite . With hypoxanthine and xanthine as substrates for growth again only the addition of selenite was necessary to show a similar increase in xanthine dehydrogenase activity . C . acidiurici could be grown in a mineral medium . Both xanthine dehydrogenase and formate dehydrogenase exhibited the highest level of activity if selenite and tungstate were present in that medium . In C . cyclindrosporum the basal activity level of xanthine dehydrogenase was about 0.95 mumol/min x mg of protein . The addition of 10(-7) M selenite to the growth medium increased the activity level about 3-fold, but the highest level (3.7 U/mg) was reached if 10(-7) M molybdate was also added . The presence of tungstate resulted in a decreased enzyme activity.

Appl Environ Microbiol, 1979 Jun, 37(6), 1127 - 31
Bacterial formation of omega-muricholic acid in rats; Sacquet EC et al.; In the feces of conventional rats, the amount of omega-muricholic and hyodeoxycholic acids vary according to the diet . To understand this phenomenon, we investigated the bacterial formation of these bile acids . The present paper reports the first isolation, from conventional rat feces, of a strain of Clostridium group III which transforms beta-muricholic acid, the main bile acid in germfree rats, into omega-muricholic acid.

Eur J Biochem, 1979 Jun, 97(1), 103 - 12
Purification and some properties of a hitherto-unknown enzyme reducing the carbon-carbon double bond of alpha, beta-unsaturated carboxylate anions; Tischer W et al.; 2-Enoate-reductase, a previously unknown soluble enzyme is present in Clostridium kluyveri and another Clostridium species growing on (E)-2-butenoate . From the latter the reductase was purified 88-fold with an overall yield up to 74% . The enzyme was pure as judged by polyacrylamide gel electrophoresis with and without sodium dodecyl sulphate as well as by isoelectric focusing . The purification of the enzyme was performed in the presence of (E)-2-methyl-2-butenoate as substrate to keep the enzyme in the oxidized state and under anaerobic conditions . The purification procedure included an ammonium sulphate precipitation, chromatography on DEAE-Sepharose CL-6B, hydroxylapatite and Sepharose CL-6B . The enzyme reduces different alpha,beta-unsaturated carboxylate anions such as (E)-2-butenoate, (E)-2-methyl-2-butenoate, (E)-cinnamate and probably many others in a NADH-dependent reaction to the saturated carboxylate anions . Fumarate, 3-phenyl-2-propinate, 2-enoyl-methyl and CoA esters proved not to be substrates for the purified reductase . NADPH does not act as an electron donor . The enzyme was shown to have a molecular weight of about 450,000 by gel chromatography . It consists of subunits with a molecular weight of 78,000 . Per subunit about 1 FAD, 3.5--3.8 atoms of iron and 4.0 labile sulphur atoms have been found indicating a conjugated iron-sulphur flavo-protein . Copper could not be detected . The isoelectric point was 8.4 . As shown by absorption spectroscopy the enzyme can be reduced by NADH and reoxidized with dichloroindophenol, hexacyanoferrate III, oxygen and substrates . Addition of 8 mol p-hydroxymercuribenzoate to 1 mol subunit completely destroyed the activity of the reductase . So far no physiological role of the enzyme is known.

Can J Microbiol, 1979 Jun, 25(6), 719 - 21
Thiosulfate formation and associated isotope effects during sulfite reduction by Clostridium pasteurianum; Chambers LA et al.; During growth of Clostridium pasteurianum on sulfite, approximately half the sulfite was reduced to sulfide and half to thiosulfate . Sulfide was enriched in 32S or 34S at different stages of growth and thiosulfate was enriched in 32S, particularly in the sulfane atom . It is suggested that thiosulfate in these bacterial cultures arose from a secondary chemical reaction . The chemical formation of thiosulfate from sulfide and sulfite was also accompanied by sulfur isotope fractionation . The implications of these results with respect to 'inverse' isotopic effects are discussed.

Can J Biochem, 1979 Jun, 57(6), 719 - 26
Structural homologies in alanine-rich acidic ribosomal proteins from procaryotes and eucaryotes; Visentin LP et al.; The amino acid composition and amino-terminal sequence have been determined for the alanine-rich, acidic ribosomal 'A' protein (equivalent to Escherichia coli L7/L12) from three procaryotic cell types that live under extreme environmental conditions (Arthrobacter glacialis, Clostridium pasteurianum, and Bacillus stearothermophilus) as well as from wheat germ, a eucaryote source . These data are compared with previously published 'A' protein sequences from other procaryotes and eucaryotes . All the procaryotic 'A' proteins, with the exception of the very acidic 'A' protein from Halobacterium cutirubrum, show similar charge, size, and amino acid composition, as well as an extensive sequence homology in the N-terminal region . Some differences are observed between gram-negative and gram-positive bacteria . The 'A' proteins from eucaryotes contain two tyrosine molecules, an amino acid absent in procaryotic 'A' proteins, as well as a reduced number of valine residues and an increased amount of aspartic acid . The N-terminal sequence of wheat germ 'A' protein shows considerable homology with other eucaryotic 'A' proteins and also with H . cutirubrum . It also shows some sequence homology with E . coli 'A' proteins.

J Urol, 1979 Jun, 121(6), 819 - 20
Emphysematous cystitis associated with Clostridium perfringens bacteremia; Maliwan N; Anaerobes are recognized rarely as the cause of urinary tract infection . A case is reported in which there were clinical signs of sepsis, positive blood culture for Clostridium perfringens and radiographic demonstration of emphysematous cystitis without any other recognized source of infection.

Am J Clin Nutr, 1979 Jun, 32(6), 1231 - 7
Comparative apparent digestibility of casein in holoxenic, axenic, and Clostridium bifermentans monoassociated rats; Corring T et al.; The aim of the present study was to determine the role of the intestinal microflora in the dietary utilization of casein . The apparent digestibility of dry matter, organic matter, energy, and nitrogen was measured in holoxenic, axenic, and Clostridium befermentans monoassociated rats deprived or not deprived of their pancreatic and bile secretions . The apparent digestibility of energy decreased in all the animals whatever the bacterial environment, after ligation of the common duct . Concerning the apparent digestibility of nitrogen, the intestinal microflora allowed the rats to compensate for the absence of the pancreatic proteolytic enzymes . It is suggested that a small part of casein, which was not hydrolyzed when the animals were deprived of their pancreatic and bile secretions, underwent a hydrolysis from the bacterial proteases.

Arch Latinoam Nutr, 1979 Jun, 29(2), 208 - 19
{Nutritional optimization of thermal processing of canned food}; Barreiro Mendez JA et al.; The model developed by Barreiro, Salas and Herrera (7) for the prediction of nutrient losses during the thermal processing of conduction heated foods was used in this work to optimize the thermal processing, maximizing nutrient retention in processes with equivalent sterilization values . The processes were stimulated in a digital computer (taking pea puree canned in cans 307 x 409 as the product analyzed) . Aall of the processes had equivalent sterilization values with respect to Clostridium botulinum . The thiamine retention associated with each process was calculated by the model of Barreiro, Salas and Herrera (7) . The maximum fraction of thiamine retained was of 0.688 for a process at 114.2 degrees C (237.5 degrees F) for 95 minutes . Also, the possibility of the existence of a can with optimum dimensions for maximum thiamine retention, with the same sterilization values, was studied . It was found that this optimum, with the same does not exist; on the contrary, the retention was at a minimum when the diameter approaches infinite and the height zero, and viceversa . For this reason, from the nutrient retention point of view, it is better to process the product in flat or slender cans for a given sterilization value.

Appl Environ Microbiol, 1979 Jun, 37(6), 1173 - 5
Enzyme-linked immunosorbent assay for detection of Clostridium botulinum type E toxin; Notermans S et al.; The enzyme-linked immunosorbent assay using the "double-sandwich" technique was utilized to determine Clostridium botulinum type E toxin . With this technique, about 80 mouse intraperitoneal 50% lethal doses of toxin could be detected . Cross-reaction was hardly observed with C . botulinum type A and B toxins . No cross-reaction was observed with culture supernatants of C . botulinum type C or other Clostridium strains . In all probability this was due to the high specificity of the antiserum prepared aginst the toxic component of type E toxin.

Appl Environ Microbiol, 1979 Jun, 37(6), 1103 - 9
Sodium nitrite and sorbic acid effects on Clostridium botulinum spore germination and total microbial growth in chicken frankfurter emulsions during temperature abuse; Sofos JN et al.; Samples of (i) a control or of (ii) sodium nitrite-containing or (iii) sorbic acid-containing, mechanically deboned chicken meat frankfurter-type emulsions inoculated with Clostridium botulinum spores, or a combination of ii and iii, were temperature abuse at 27 degrees C . Spore germination and total microbial growth were followed and examined at specified times and until toxic samples were detected . The spores germinated within 3 days in both control and nitrite (20, 40 and 156 micrograms/g) treatments . Sorbic acid (0.2%) alone or in combination with nitrite (20, 40, and 156 micrograms/g) significantly (P less than 0.05) inhibited spore germinations . No significant germination was recorded until toxic samples were detected . A much longer incubation period was necessary for toxin to be formed in nitrite-sorbic acid combination treatments as contrasted with controls or nitrite and sorbic acid used individually . Total growth was not affected by the presence of nitrite, whereas sorbic acid appeared to depress it . Possible mechanisms explaining the effects of nitrite and sorbic acid on spore germination and growth are postulated.

Zentralbl Bakteriol {Orig A}, 1979 Jun, 243(4), 528 - 41
{Experimental studies on the pathogenicity of tetanus spores regarding eucaryotic cell systems (author's transl)}; Schneeweiss U et al.; After injection of known numbers of viable tumour cells admixed with tetanus spores into the subcutaneous tissue between the shoulder blades the sigmoidal tetanus lethality curves of mice can be quantitatively analysed following the underlying growth of tetanus clostridia and tumour cells . We suggest that the "driving force" for exponential clostridial growth and toxin synthesis are oxygen-consuming clones of tumour cells . Our experimental data are in good accordance with a stochastic mathematical model which allows for computing the cloning probability of one single spore on the basis of a 1-mitosis-1-clostridium division-principle.

Appl Environ Microbiol, 1979 Jun, 37(6), 1196 - 200
Raffinose increases sporulation and enterotoxin production by Clostridium perfringens type A; Labbe RG et al.; Replacement of starch with raffinose in Duncan and Strong sporulation medium improved percent sporulation in six of eight strains tested . Enterotoxin concentration in cell extracts was increased in the case of four of five known enterotoxin-positive strains . With strain NCTC 10240, levels of 0.3, 0.4, and 0.5% raffinose produced the highest enterotoxin concentration 300 to 320 micrograms of enterotoxin per mg of cell extract protein . At a level of 0.4% raffinose the highest enterotoxin concentration in cell extracts of NCTC 10240 occurred after 8 h of growth in Duncan and Strong medium . Enterotoxin produced in the presence of starch or raffinose by three separate strains all migrated at similar Rm by polyacrylamide gel electrophoresis.

Am J Vet Res, 1979 Jun, 40(6), 876 - 81
Occurrence of anaerobic bacteria in diseases of the dog and cat; Berg JN et al.; A survey for anaerobic bacteria was conducted in 314 clinical specimens from dogs and cats . A total of 187 anaerobic isolates in pure and mixed culture were isolated from 111 of the specimens that contained anaerobic bacteria . Common isolated included Actinomyces (9.1%), Clostridium perfringens (19.3%), other Clostridium spp (11.2%), Peptostreptococcus anaerobius (7.5%), Bacteroides melaninogenicus (13.4%), other Bacteroides spp (17.6%), and Fusobacterium necrophorum (5.3%) . Anaerobic bacteria were involved in serious lesions that often were life threatening to the animals . Antibiotic susceptibility data indicated that the lincomycin family, the penicillin family, chloramphenicol, and cephaloridine are preferred drugs for treatment of anaerobic infections . Data from the survey were used in formulation of a table to aid practitioners in clinical diagnosis of disease caused by anaerobes . Clostridium perfringens was isolated in large numbers from five of six dogs with a clinical diagnosis of canine hemorrhagic gastroenteritis and from one cat with hemorrhagic diarrhea . Experimental infections were induced in rats, using caine feces as inoculum . Induced lesions contained aerobic and anaerobic bacteria similar to those bacteria isolated in the clinical survey, indicating that feces may serve as a major source of these bacteria in clinical infections of the dog.

J Gen Microbiol, 1979 Jun, 112(2), 225 - 33
The effect of transition metal ions on the resistance of bacterial spores to hydrogen peroxide and to heat; Waites WM et al.; The presence of 10 microM-Cu2+ increased the lethal effect of hydrogen peroxide on spores of Clostridium bifermentans but not on those of Clostridium sporogenes PA 3679, Clostridium perfringens, Bacillus cereus or Bacillus subtilis var . niger . Cu2+ at 100 muM also increased the lethal effect of heat on spores of C . bifermentans but not on those of B . sutilis var . niger . The rate and extent of Cu2+ uptake by spores of C . bifermentans and B . subtilis var . niger were similar, but examination of unstained sections of spores by electron microscopy suggested that Cu2+ is bound by the protoplasts of spores of C . bifermentans but not of B . subtilis var . niger.

Biochemistry, 1979 May 29, 18(11), 2335 - 9
Mechanism of Lactobacillus leichmannii ribonucleotide reductase studied with Coalpha-{alpha-(Aden-9-yl)}-Cobeta-adenosylcobamide (Pseudocoenzyme B12) as coenzyme; Blakley RL et al.; Coalpha-{alpha-(Aden-9-yl)}-Cobeta-adenosylcobamide (pseudocoenzyme B12) purified from Clostridium tetanomorphum has been reacted with ribonucleotide reductase purified from Lactobacillus leichmannii under various conditions, and the properties of the products obtained have been compared by electron paramagnetic resonance (EPR) with those previously reported for products formed from the normal coenzyme (adenosylcobalamin) . The rapidly formed intermediate and the slowly formed "doublet" species from the pseudocoenzyme have EPR spectra identical with those formed from the normal coenzyme . This and other considerations make it less likely that the unusual magnetic properties of the rapidly formed intermediate are due to strongly distorted octahedral symmetry about Co(II) as previously postulated . Instead it is probable that the EPR spectrum is due to interaction of the radical pair by both exchange coupling and magnetic dipole--dipole coupling . Although Coalpha-{alpha-(aden-9-YL)}cob(II)amide in solution does not show superhyperfine splitting in the EPR spectrum because of its base-off configuration, the cob(II)amide formed by degradation of the pseudocoenzyme within the catalytic site of the enzyme did show triplets due to a nitrogen axially coordinated to cobalt . This suggests that binding of the cob(II)amide to the reductase catalytic site causes a shift to the base-on form.

Biochim Biophys Acta, 1979 May 25, 573(2), 332 - 42
Transformation of 4-androsten-3,17-dione by growing cultures and cell extracts of Clostridium paraputrificum; Glass TL et al.; Growing cultures of Clostridium paraputrificum transformed 4-androsten-3,17-dione to 3 alpha-hydroxy-5 beta-androstan-17-one in a sequential manner with 5 beta-androstan-3,17-dione as an intermediate . The addition of 1.5 mM menadione to log-phase cultures which had formed 5 beta-androstan-3,17-dione resulted in a partial reoxidation of this steroid to 4-androsten-3,17-dione . However, this treatment also resulted in transient inhibition of culture growth . Resumption of growth was accompanied by complete reduction of 4-androsten-3,17-dione to 5 beta-androstan-3,17-dione . Cell extracts of C . paraputrificum were capable of carrying out these reductive transformations in the absence of added cofactors . However, Sephadex G-25 treated extracts required NADH or NADPH for these reactions . A flavin nucleotide, either FAD (plus NADH or NADPH) or FMN (plus NADH) was highly stimulatory for 4-androsten-3,17-dione reduction to 5 beta-androstan-3,17-dione . NADH was the preferred reduced pyridine nucleotide for reduction of the C4-C5 double bond, while time-course measurements suggested that NADPH was the preferred donor for reduction of the 3-keto group.

J Membr Biol, 1979 May 25, 47(3), 285 - 301
Effect of purified phospholipases on the binding of tetrodotoxin to axon plasma membrane; Chacko GK; The role of phospholipids in the binding of {3H}tetrodotoxin to garfish olfactory nerve axon plasma membrane was studied by the use of purified phospholipases . Treatment of the membranes with low concentrations of either phospholipase A2 (Crotalus adamanteus and Naja naja) or phospholipase C (Bacillus cereus and Clostridium perfringens) resulted in a marked reduction in tetrodotoxin binding activity . A 90% reduction in the activity occurred with about 45% hydrolysis of membrane phospholipids by phospholipase A2, and with phospholipase C the lipid hydrolysis was about 60--70% for a 70--80% reduction in the binding activity . Phospholipase C from B . cereus and Cl . perfringens had similar inhibitory effects . Bovine serum albumin protected the tetrodotoxin binding activity of the membrane from the inhibitory effect of phospholipase A2 but not from that of phospholipase C . In the presence of albumin about 25% of the membrane phospholipids remained unhydrolyzed by phospholipase A2 . It is suggested that these unhydrolyzed phospholipids are in a physical state different from the rest of the membrane phospholipids and that these include the phospholipids which are directly related to the tetrodotoxin binding component . It is concluded that phospholipids form an integral part of the tetrodotoxin binding component of the axon membrane and that the phospholipase-caused inhibition of the binding activity is due to effects resulting from alteration of the phospholipid components.

Arch Dermatol, 1979 May, 115(5), 580 - 1
Induction of colitis in hamsters by topical application of antibiotics; Feingold DS et al.; Syrian hamsters are exquisitely sensitive to clindamycin; as little as 1 mg/kg of clindamycin given systemically causes a fatal colitis . Clindamycin and erythromycin were applied topically daily to the shaved backs of Syrian hamsters in a hydroalcoholic vehicle . A daily dose of 0.1 mg of clindamycin was lethal to more than half the hamsters and 1 mg to all the animals . The antibiotic-associated toxin from Clostridium difficile was present in their cecal material . Based on body surface areas and estimated usual volumes applied, the lethal dose in hamsters is not dissimilar to that given humans for acne . Oral tetracycline therapy protected the animals from clindamycin toxicity, but the animals died three days after stopping tetracycline if topical clindamycin applications were continued.

J Clin Microbiol, 1979 May, 9(5), 627 - 8
Survival of anaerobic bacteria in common laboratory diluents; Casciato DA et al.; The survival of six species of anaerobic bacteria was studied in simple or commercially available diluents . Bacteroides fragilis and Fusobacterium nucleatum showed excellent survival in all diluents including distilled water . Fusobacterium mortiferum survived well in all diluents except water and water supplemented with 0.1% gelatain . Clostridium perfringens survived best in phosphate-buffered saline with gelatin . Peptococcus asaccharolyticus required gelatin added to the basic diluent, and Streptococcus intermedius showed excellent survival only in minimal essential medium with gelatin . These diluents could provide effective and economical alternatives to more complex and costly diluents often used in work with anaerobic bacteria.

Appl Environ Microbiol, 1979 May, 37(5), 978 - 84
Effect of reducing agents on oxidation-reduction potential and the outgrowth of Clostridium botulinum type E spores; Smith MV et al.; Oxidation-reduction potential (Eh) levels were measured and standardized to pH (Eh7) for Trypticase soy broth containing various concentrations of reducing agents . Prereduced Trypticase soy broth with no added reducing agents exhibited a potential of -141 mV . Ascorbic acid at 0.2 to 0.005% and sodium thioglycolate at concentrations below 0.05% produced an Eh7 higher than the prereduced Trypticase soy broth containing no added reducing agents . The addition of cysteine hydrochloride,2-mercaptoethanol, and sodium formaldehyde sulfoxylate to prereduced Trypticase soy broth resulted in a reduction of Eh7 compared to the system without added reducing agents . The order of relative reducing intensity (from highest to lowest) for the reducing agents when comparing molar concentration was: sodium formaldehyde sulfoxylate,2-mercaptoethanol, cysteine hydrochloride, sodium thioglycolate, and ascorbic acid . Optimal growth of the test organism occurred at low Eh7 and low concentration of the reducing agents . A direct correlation existed between growth of the test organism and -Eh7 x -log concentration of the reducing agent.

J Gen Microbiol, 1979 May, 112(1), 61 - 6
Host modification of chlamydiae: presence of an egg antigen on the surface of chlamydiae grown in the chick embryo; Allan I et al.; Egg-grown chlamydiae (EGO) have a yolk sac antigen assoicated with their surface which is absent from cell monolayer-grown organisms (CGO) . EGO infectivity was specifically neutralized by rabbit antiserum to normal yolk sac; CGO infectivity, before or after incubation with normal yolk sac material, was not neutralized . Treatment of EGO with Clostridium welchii culture filtrate, containing phospholipase C, abolished spontaneous infectivity for monolayers and neutralization by anti-yolk sac antiserum but did not affect centrifuge-assisted infectivity . The possible significance of host antigen on the chlamydial surface is considered.

J Gen Microbiol, 1979 May, 112(1), 203 - 6
High and low toxin production by a non-toxigenic strain of Clostridium botulinum type C following infection with type C phages of different passage history; Oguma K et al.; Toxin production in Clostridium botulinum types C and D is governed by specific bacteriophages . Prior passages of a phage controlling type C toxin production caused subsequently lysogenized bacteria to become variably toxigenic . This appears to be one of the causes of the decrease in toxigenicity which is common in some type C and D strains . The morphology of bacteria was also changed from rod-shaped to filamentous by infection with a successively propagated phage.

Nord Vet Med, 1979 May, 31(5), 214 - 21
Clostridium botulinum in fish; Huss HH et al.; 1407 fish caught in Scandinavian waters, the North Sea and the North Atlantic have been examined for the presence of Cl . botulinum . The incidence in gut samples expressed as percentage of fish tested was generally highest in fish from Scandinavian coastal waters and the Baltic Sea (4--43%), decreasing in fish from the North Sea (0--8%), and the organism was practically absent in fish from the North Atlantic . When gut samples were examined, the incidence was highest in demersal fish (cod and flatfish) as compared with pelagic fish (herring) . The latter fish species were mainly contaminated on outer surfaces and gills . Only type E was detected in this survey, and Cl . botulinum was not detected in any wild fresh water fish . It is suggested that type E spores may originate in the sea bed and that they be spread by fish and water currents.

Ann Microbiol (Paris), 1979 May-Jun, 130 A(4), 441 - 8
{Presence of antibodies in human colostral secretory IgA against enteric commensal bacteria: biological implications (author's transl)}; Liem ND et al.; Antisecretory component, anti-alpha, anti-mu and anti-Fc (gamma) fluorescent antibodies were used to detect the presence of immunoglobulins with antibody activity against enteric commensal bacteria in human colostrum and serum . Forty nine colostrum samples were studied; all of them displayed secretory IgA (sIgA) antibodies reacting with Bacteroides thetaiotaomicron, Clostridium perfringens and Escherichia coli serotype O141:H32 without any K antigen . The amount of sIgA antibodies was always related to the sIgA colostral concentration varying greatly from one patient to another . For the 3 lactating women studied, the colostrum sIgA antibodies were largely predominant as compared to the antibodies of other classes; in their sera, no antibody having the same anticommensal specificity was detected in the IgA fraction while these antibodies were found in IgM and IgG . Our results are incompatible with the existence of local antigenic stimulation, and the IgA transfer from serum into mammary secretion appears unlikely, but these results are perfectly compatible with the antigenic stimulation of gut associated lymphoid tissue and subsequent migration in mammary tissue.

Appl Environ Microbiol, 1979 May, 37(5), 1038 - 40
Effect of heat treatment on the performance of tryptose-sulfite-cycloserine agar for enumeration of Clostridium perfringens; Brodsky MH et al.; Dissolving dehydrated tryptose-sulfite-cycloserine agar by only boiling or microwaving was found to inhibit Clostridium perfringens colony development in pour plates when compared with C . perfringens recovery in tryptose-sulfite-cycloserine agar prepared by autoclaving.

Can J Microbiol, 1979 May, 25(5), 642 - 5
High yields of coatless spores of Clostridium perfringens strain 8--6 in a defined medium; Sacks LE et al.; Coatless spores of mutant strain 8--6 of Clostridium perfringens were formed reproducibly at levels exceeding 10(7)/mL in a defined medium . Confirming previous observations, the spores are quite stable in water and resistant to heat, octanol, and ethanol . They also resist 5% phenol . Electron microscopy confirms, with further evidence, the probable absence of any structure exterior to the cortex . The levels of sporulation attained are sufficient to permit recovery and purification for future studies on physicochemical and germination properties . The defined medium makes possible studies on spore coat (and exterotoxin) biosynthesis and coat assembly.

J Cell Physiol, 1979 May, 99(2), 191 - 200
The effects of Clostridium perfringens enterotoxin on morphology, viability, and macromolecular synthesis in Vero cells; McClane BA et al.; Vero (African green monkey kidney) cells grown in tissue culture monolayer were sensitive to Clostridium perfringens enterotoxin . Within 30 minutes of exposure to the enterotoxin gross morphological damage was observed and within 40 minutes approximately 75% of the cells had detached . Nearly half of the cells were nonviable following 35 to 40 minutes incubation with the enterotoxin . Doses as low as 0.1 ng caused small but detectable inhibition of plating efficiency of the cells while more than 100 ng caused the inhibition to approach 100% . Total inhibition of DNA, RNA, and protein synthesis occurred within 30 minutes exposure to enterotoxin . Heat inactivated enterotoxin had no apparent effects upon cellular morphology, detachment, viability, plating efficiency, or incorporation . We propose that the enterotoxin induces structural damage to the cytoplasmic membrane which results in loss of electrolytes and other essential substances from the cells . The outcome of this process is shut down of macromolecular synthesis, gross morphological damage, and eventual death of the cell.

J Infect Dis, 1979 May, 139(5), 586 - 9
Ornidazole and anaerobic bacteria: in vitro sensitivity and effects on wound infections after appendectomy; Palmu A et al.; The sensitivities of 68 clinical isolates of Bacteroides fragilis, 18 of Clostridium perfringens, and 11 of other Clostridium species were tested against ornidazole alone and in combination with ampicillin and gentamicin . A concentration of 3.1 microgram of ornidazole/ml inhibited 98% of the strains of B . fragilis, with greater sensitivity when ampicillin and gentamicin were also present . A concentration of 6.2 microgram of ornidazole/ml inhibited 16 of 18 strains of C . perfringens and all 11 strains of other Clostridium species . Concentrations in serum and tissue were determined after intravenous infusion of 500 mg of ornidazole 15 min prior to appendectomy . During the operation the concentration in serum was 7.90 +/- 0.57 microgram/ml, and in appendix tissue, 5.26 +/- 0.60 microgram/g . In the series of 200 patients undergoing appendectomy, six patients treated with ornidazole and 12 patients treated with placebo developed a wound infection . In patients with perforated appendix, the rate of wound infection was 7.1% in those given ornidazole and 63.6% in those given placebo (P = 0.004) . Not a single B . fragilis was isolated from appendix swabs or wound exudates after prophylaxis with ornidazole.

J Biol Chem, 1979 Apr 25, 254(8), 2626 - 9
Bacterial metabolism of corticoids with particular reference to the 21-dehydroxylation; Winter J et al.; Clostridium paraputrificum, an obligate anaerobe recovered from human feces, reduces the alpha,beta-unsaturated carbonyl of deoxycorticosterone, corticosterone, cortisone, and cortisol . The same steroids are 21-dehydroxylated by Culture 116, recently isolated from human feces, and by a closely related organism, Eubacterium lentum . The 21-dehydroxylase has no effect on hydroxyl groups at carbon atoms 11 and 17.

J Biol Chem, 1979 Apr 25, 254(8), 3045 - 53
Inhibitor of human collagenase from cultures of human tendon; Vater CA et al.; A potent inhibitor of human collagenases, released from human tendon explants in culture, has been purified and partially characterized . The tendon inhibitor has an estimated molecular weight of 25,000 . It is relatively heat-stable but undergoes loss of activity following exposure to trypsin . It inhibits trypsin-activated rheumatoid synovial collagenase as well as the enzyme obtained from polymorphonuclear leukocytes . No inhibition of collagenase from Clostridium histolyticum (clostridiopeptidase A, EC 3.4.24.3) was noted . This collagenase inhibitor may be a factor in the regulation of extracellular connective tissue catabolism.

Biochim Biophys Acta, 1979 Apr 12, 567(2), 453 - 63
Carbamyl phosphate-dependent ATP synthesis catalyzed by formyltetrahydrofolate synthetase; Buttlaire DH et al.; Formyltetrahydrofolate synthetase (formate:tetrahydrofolate ligase (ADP-forming), EC 6.3.4.3) from Clostridium cylindrosporum catalyzes phosphate transfer from carbamyl phosphate to ADP . This activity is lost when monovalent cations are removed and is recovered when K+ is added back . Carbamyl phosphate is an inhibitor of the formyltetrahydrolfolate synthetase forward reaction, and formate as well as phosphate inhibit the ATP synthesis reaction . Acetyl phosphate and phosphonoacetate are inhibitors of both reactions . The results of kinetic studies support the concept that carbamyl phosphate is an analog of the putative intermediate of the formyltetrahydrofolate synthetase reaction, formyl phosphate.

Ann Trop Med Parasitol, 1979 Apr, 73(2), 145 - 8
Outbreak of botulism in Kenyan nomads; Smith DH et al.; During disease surveillance in Kenya, a series of deaths were investigated among a group of nomadic Gabra in Marsabit . The cause was identified as botulism (Clostridium botulinum Type A), contracted from sour milk prepared traditionally in a gourd . Reported outbreaks of botulism in Africa would appear to be extremely rare.

Nucleic Acids Res, 1979 Apr, 6(4), 1449 - 65
Core nucleosomes by digestion of reconstructed histone-DNA complexes; Bryan PN et al.; Reconstructed complexes of the inner histones (H2A, H2B, H3, H4) and a variety of DNAs were digested with micrococcal nuclease to yield very homogeneous populations of core nucleosomes (nu 1) . Nucleosomes containing Micrococcus luteus DNA (72% G+C); chicken DNA (43% G+C), Clostridium perfringens DNA (29% G+C); or poly(A-dT.poly(dA-dT) have been examined by circular dichroism, thermaldetenaturation, electron microscopy, and DNAse I digestion . Circular dichroism spectra of all particles show a typically suppressed ellipticity at 260--280 nm and a prominent alpha-helix signal at 222 nm . All particles show biphasic melting except nu 1 (dA-dT), which show three prominent melting transitions at ionic strength less than or equal to 1 mM . DNAse I digestion of nu 1 (dA-dT) produces a ladder of DNA fragments fiffering in lengthy by one base residue . nu 1 (dA-dT) contain 146 base pairs of DNA and exhibit an average DNA helix pitch of 10.4-10.5 bases per turn . There appear to be two regions of different DNA pitch wihtin nu 1 (dA-dT) . It is suggested that the two regions of DNA pitch might correspond to the two regions of the melting profiles.

Crit Care Med, 1979 Apr, 7(4), 176 - 81
Tetanus: a review; Alfery DD et al.; Tetanus is caused by the organism Clostridium tetani, which produces tetanospasmin, a neurotoxin responsible for the clinical manifestations of muscle rigidity and reflex spasms . The majority of cases follow an anaerobic wound infection associated with trauma . Incubation period is usually 3 days to 3 weeks . 75% of patients present with trismus . Reflex spasms are seen in 70% of patients and characterize the severity of the disease . Treatment involves removal of the offending organism, neutralization of free neurotoxin, controlling rigidity and reflex spasm, and minimizing complications . Diazepam may be used alone in mild cases . Severe cases require the addition of nondepolarizing neuromuscular blocking agents and mechanical ventilation . Respiratory complications occur early and require aggressive airway management . A serious, late complication is the syndrome of sympathetic nervous system overactivity that is treated with alpha and beta blockade . High mortality rates seen in the United States may be due to delays in diagnosis and lack of familiarity with treatment . The disease is preventable with adequate immunization.

Can J Microbiol, 1979 Apr, 25(4), 522 - 7
Outgrowth and sporulation studies on Clostridium botulinum type E: influence of isoleucine; Hawirko RZ et al.; A defined medium (CDM) is described which supported growth and sporulation of type E strains of Clostridium botulinum, but not sporulation of other serotypes of C . botulinum or C . sporogenes . As compared to growth in complex medium, spore outgrowth was delayed and both the growth rate and the cell yield was reduced . However, efficiency of sporulation of the type E MSpt strain in a chemically defined medium (CDM) was the same as that in complex medium and, in fact, sporulation was nearly synchronous and completed within 3 h of the first appearance of phase-bright endospores, compared with completion in 9 h in TPGY . Growth studies with CDM, from which single amino acids were omitted, showed that isoleucine was essential for outgrowth of heat-activated spores of the MSp+ strain, whereas valine was required for that of the Ts-25 mutant . Radioactive isoleucine was incorporated by germinating MSp+ spores at an earlier stage and at a more rapid rate than labelled methionine or mixed amino acids . Uptake studies showed that isoleucine accumulated in a prominent acid-soluble pool during outgrowth, a period when its incorporation into protein was not evident . The results suggest that the isoleucine may be required for a purpose other than protein synthesis during outgrowth.

J Cell Biol, 1979 Apr, 81(1), 43 - 9
Binding of Clostridium botulinum neurotoxin to the presynaptic membrane in the central nervous system; Hirokawa N et al.; Large synaptosome fractions were isolated from the cerebellar and cerebral cortices of rats and were incubated with Clostridium botulinum type A neurotoxin in vitro . The binding of the neurotoxin to the synapses was observed by electron microscopy, using the double-sandwich immunocytochemical method . Botulinum neurotoxin was preferentially bound to the presynaptic membrane in the large synaptosome fraction . The binding regions for the neurotoxin were localized on both the extrajunctional and junctional areas of the presynaptic membranes and appeared as patches of various sizes . However, they did not exist on the postsynaptic membranes . Botulinum neurotoxin is proposed to be a useful analytical tool for understanding the characteristics of the presynaptic membranes in the central nervous system.

Infect Immun, 1979 Apr, 24(1), 7 - 11
Experimental reproduction of neonatal diarrhea in young gnotobiotic hares simultaneously associated with Clostridium difficile and other Clostridium strains; Dabard J et al.; Clostridium difficile, C . perfringens, and C . tertium are very often present simultaneously in the feces of conventional diarrheic young hares, whereas these three bacterial species are rarely encountered and never present simultaneously in the feces of healthy young hares . When a strain of each of the three bacterial species was monoassociated with axenic young hares, the appearance of pathological disorders was only observed in animals monoassociated with C . difficile, when the number of C . difficile exceeded 10(8) per g of fresh feces . When a strain of C . perfringens or a strain of C . tertium, or both, was associated with C . difficile, diarrhea and death occurred more rapidly than in hares monoassociated with C . difficile . C . difficile and C . perfringens became established more rapidly when disassociated than when monoassociated with axenic hares . The association of C . perfringens and C . tertium with axenic hares did not bring about any pathological disorders . It may be concluded that C . difficile is the causal agent of neonatal diarrhea in conventional and gnotobiotic young hares and that other strains of Clostridium enhance its pathogenic effect . C difficile alone or associated with C . perfringens or C . tertium does not play any pathogenic role in young rats, mice, or rabbits.

Appl Environ Microbiol, 1979 Apr, 37(4), 667 - 9
Survival of bacteria in carcasses; Gill CO et al.; Bacteria injected into the bloodstream of guinea pigs shortly before death decreased in number in carcass tissues for about 1 h after death . If initial bacterial numbers were sufficiently low, all bacteria were eliminated, and carcass tissues were sterile 24 h after death . Carcass tissue sterility was maintained with an initial density of Clostridium perfringens or Salmonella typhimurium of 20 cells per g or with an initial density of the other species examined of several hundred cells per gram . With larger numbers of strict and facultative anaerobes, growth commenced after 3 h in carcasses incubated at 30 degrees C . Spores of C . perfringens were killed over the same period as vegetative cells, but growth did not commence until 8 h after death . Bactericidal activity in carcass tissues must therefore be taken into account in evaluating the significance of reports of deep-tissue contamination of carcasses from meat animals.

Arch Surg, 1979 Apr, 114(4), 445 - 8
Intestinal strangulation in germfree and monocontaminated dogs; Yale CE et al.; Earlier studies in the germfree rat demonstrated that the common intestinal bacteria vary greatly in their ability to cause death after intestinal strangulation . Some of these experiments were repeated in adult, germfree and monocontaminated beagles . Neither short closed-loop hemorrhagic nor long closed-loop ischemic strangulation killed the germfree dog . Either procedure rapidly killed the dog with a conventional bacterial flora or a dog monocontaminated with Clostridium perfringens . The dogs monocontaminated with Bacteroides fragilis died after several days, whereas Escherichia coli killed only one of three animals . These experiments demonstrate the usefulness of the germfree dog as a unique research subject, confirm our earlier studies on intestinal strangulation in germfree rats, and further emphasize the differing lethal potentials of the intestinal bacteria in intestinal strangulation.

Arch Ophthalmol, 1979 Apr, 97(4), 661 - 3
Clostridium perfringens corneal ulcer; Stern GA et al.; A corneal ulcer caused by Clostridium perfringens developed in a 76-year-old woman with Sjogren's syndrome . Experimental C perfringens keratitis was induced in rabbits by the intrastromal injection of 10(7) organisms . In both our patient and the experimental animals, a bullous lesion overlay the affected area of the cornea . This may be a specific lesion in clostridial infections of the cornea . Clostridium perfringens should be regarded as an opportunistic corneal pathogen, and anaerobic cultures should be performed in all cases of suspected bacterial corneal ulcer.

Med Microbiol Immunol (Berl), 1979 Mar 13, 167(1), 61 - 70
Metronidazole bioassay with increased sensitivity; Jokipii L et al.; The bioassay of metronidazole using Clostridium butyricum incorporated in agar plates detected concentrations higher than 1.0 microgram/ml . Gentamicin in the sample did not affect the growth of the target organism or the inhibition by metronidazole . Penicillin in the sample could be eliminated by the incorporation of penicillinase in the agar or by using as the target organism a surface inoculum of the penicillin-resistant Bacteroides fragilis . Increasing duration and temperature of aerobic prediffusion before allowing the growth of the strictly anaerobic Cl . butyricum increased the diameters of inhibition by metronidazole, but did not affect the threshold of detectability . The incorporation of metronidazole in the agar increased the sensitivity of the assay ten- to 100-fold.

J Biol Chem, 1979 Mar 10, 254(5), 1462 - 8
alpha-Clostripain . Chemical characterization, activity, and thiol content of the highly active form of clostripain; Gilles AM et al.; A highly active form of clostripain, composed of two polypeptide chains (Mr = 43,000 and 12,500), was isolated by hydrophobic chromatography from the culture medium of Clostridium histolyticum . It differs in amino acid composition, namely in the value for cyst(e)ine, from that previously reported . The analyses of the separated chains are given . Activity is related to the number of free cysteine residues and full activity is obtained only after complete reduction of the disulfide bonds . Specific modifications by sulfhydryl reagents and tosyl lysine chloromethyl ketone of one thiol group, the one implicated in the activity, are reported . High specificity of alpha-clostripain is restricted to arginyl peptide bonds as tested on parvalbumin.

Rev Infect Dis, 1979 Mar-Apr, 1(2), 386 - 97
Antibiotic-associated colitis: effects of antibiotics on Clostridium difficile and the disease in hamsters; Fekety R et al.; Fifteen isolates of Clostridium difficile from hamsters and human patients were inhibited or killed by low concentrations of metronidazole, vancomycin, penicillin, and ampicillin; the isolates were often reesistant to tetracycline, cephalosporins, trimethoprim-sulfamethoxazole, clindamycin, erythromycin, and aminoglycosides . Antibiotics to which C . difficile was susceptible were able to prevent or postpone the colitis caused by clindamycin in hamsters . Colitis could be produced by treatment of hamsters with any one of these antibiotics . Production of colitis not only involved selection of resistant variants, but in some instances seemed to result from the acquisition of organisms after treatment, their persistence despite treatment, or from subinhibitory cecal concentrations of antibiotic (explainable by either pharmacologic factors or enzymatic inactivation) . As in humans, no organisms other than C . difficile have been implicated conclusively as etiologic agents of colitis in hamsters . Our results suggest it may be wise to use isolation precautions for patients with colitis caused by C . difficile.

Rev Infect Dis, 1979 Mar-Apr, 1(2), 379 - 85
Partial purification and characterization of a cytotoxin from Clostridium difficile; Taylor NS et al.; A trypsin-sensitive, heat-labile cytotoxin was purified from the supernatant of a culture of Clostridium difficile by a procedure that included ultrafiltration, precipitation with (NH4)2SO4, gel filtration, and ion-exchange chromatography . The procedure resulted in recovery of 20% of the cytotoxin and an estimated 1,500-fold increase in cytotoxic activity . The minimal amount of protein required to give an actinomorphic response in WI-38 cell cultures was 1.4 ng/ml . The estimated molecular weight of the cytotoxin is 240,000 . A cytotoxin having similar properties was purified from the stool of a patient with antibiotic-associated pseudomembranous colitis by (NH4)2SO4 precipitation and gel filtration chromatography . This procedure resulted in a recovery of 26% of the cytotoxin, a 50-fold increase in cytotoxic activity, and a cytotoxic response with a minimum of 12.1 ng of protein/ml.

Rev Infect Dis, 1979 Mar-Apr, 1(2), 370 - 8
Colitis induced by Clostridium difficile; Bartlett JG et al.; Clostridium difficile has been implicated as the major cause of antibiotic-associated pseudomembranous colitis . The current laboratory diagnostic test of choice is a tissue culture assay that demonstrates the presence of a cytopathic toxin neutralized by antitoxin to Clostridium sordellii . This toxin was found in stools from 42 of 43 patients with antibiotic-associated pseudomembranous colitis and in stools from 12 of 78 patients with antibiotic-associated diarrhea . Specimens from patients with gastrointestinal conditions unrelated to administration of antibiotics and those from healthy controls were uniformly negative . Neutralization of toxin by antitoxin to C . sordellii appears to represent antigenic cross-reactivity, since broth cultures of C . difficile also contain a cytopathic toxin neutralized by this antitoxin . Strains of C . difficile are susceptible to vancomycin, and the initial clinical experience with oral administration of this agent shows promising results.

Zentralbl Bakteriol {B}, 1979 Mar, 168(2), 18 - 36
{Germinal change and its significance in the hospital (author's transl)}; Alexander M; To receive objective informations about clinically supposed difference of bacteria spectrum in hospital infections within the last 20 years, we made statistical comparatively investigations between varying periods and in different fields . In the whole material of the Medical Department we found an increase of Proteus, Pseudomonas and Klebsiella (p = 0,01%) and an increase of Candida while Staphylococci, Streptococci, Pneumococci, Clostridium and Meningococci have clearly decreased . In the Department of Nephrology we found a significant increase of Proteus and Pseudomonas (in regard to the manifold findings also of Klebsiella) and a significant decrease of Enterococci (in regard to the manifold findings also of E . coli) . In the Reanimation Department there were in opposite to a strong increase of Candida, followed by Pseudomonas, Klebsiella, Enterobacter and Pneumococci, a decrease of Staphylococcus aureus, Enterococci and Streptococci in the sputum findings . In the Urological Clinic while the period of report Pseudomonas and Enterobacter increased significantly, Staphlococcus aureus and Streptococci decreased significantly . In the Neurosurgical Clinic the part of the gramnegative germs of all germinal isolations increased from 8,3% to 51,5% at which especially E . coli, Enterobacter and Pseudomonas were more often found.

Rev Chir Oncol Radiol O R L Oftalmol Stomatol Chir, 1979 Mar-Apr, 28(2), 129 - 36
{Surgical infections with anaerobic bacteria (splenic abscess ruptured into the peritoneum}; Radulescu D; A case is presented, of a patient aged 19, who, in the course of otic suppuration treated by tetracycline administration, developed a septicemic condition with a gigantic splenic abscess followed by generalized peritonitis . The initially unfavourable evolution had an improved course following splenectomy and peritoneal drainage, especially after the identification of the anaerobic germs that determined the infection (Bacteroides clostridii formis and Clostridium bifermentans) and the introduction of an adequate therapy.

Infect Immun, 1979 Mar, 23(3), 795 - 8
Ultrastructural changes of cultured human amnion cells by Clostridiu difficile toxin; Chang TW et al.; The ultrastructure of the surface of primary human amnion monolayer cells undergoing cytopathology induced by Clostridium difficile toxin was examined by scanning electron microscopy . Our observations indicated that the type and distribution of cell surface projections were altered dramatically by this toxin . The patterns of such surface changes were specific for the two different types of cells found in this cell culture . Cells with demarcated borders showed rearrangement of microvilli into globular chains or ridges which lined up with the branching membrane . Cells without demarcated borders exhibited studlike microvilli, all arranged into ridges or globular chains . These changes were noted after 1 h of toxin exposure and persisted without further progression, in spite of continued toxin exposure, up to 48 h . These data indicate that C . difficile produces a cytolytic toxin and that scanning electron microscopy may be useful in determining toxin-cell interactions.

Zentralbl Bakteriol {Orig A}, 1979 Mar, 243(1), 113 - 8
{Enzymatic mechanisms of the oncolysis by Clostridium oncolyticum M 55 ATCC 13.732 (author's transl)}; Brantner H et al.; After a short review of older attempts at explanation of the oncolysis by Cl . oncolyticum M 55 a new theory will be presented . In connection with new results of enzymological investigations the metabolic correlations between the tumor cell and the vegetative clostridial cell and the consequences of the results for an oncolytic therapy will be demonstrated.

P N G Med J, 1979 Mar, 22(1), 57 - 9
Clostridium welchii type C antitoxin in the treatment of "pig bel" (enteritis necroticans): a controlled trial in Papua New Guinea; Rooney J et al.; Enteritis necroticans remains a serious problem in the Highlands of Papua New Guinea . Earlier claims that Clostridium Welchii Type C antitoxin is effective in lowering mortality rates were subjected to a prospective controlled trial in 1972-73 . Use of the antitoxin was found to be of no benefit in treatment of this disease.

P N G Med J, 1979 Mar, 22(1), 30 - 4
The prevention of pig-bel in Papua New Guinea; Lawrence G et al.; In many Highlands hospitals pig-bel is the commonest cause of death in children over 12 months of age . In a double blind controlled trial in Sina Sina we have shown that Clostridium welchii type C beta toxoid (beta toxoid) protects against pig-bel (p < 0.02) . Of 2,538 children given beta toxoid, two developed pig-bel in the following two years . Seventeen of 2,532 control children vaccinated with tetanus toxoid developed pig-bel . We suggest that beta toxoid be given to Highlands children at two, four and six months of age . Boosters may be needed . Health services in the Highlands need to be improved so that this vaccine can be effectively delivered.

Rev Infect Dis, 1979 Mar-Apr, 1(2), 254 - 62
Virulence factors of Clostridium perfringens; Smith LD; Clostridium perfringens produces a variety of virulence factors . The mechanism of action of these factors usually falls into one of three groups . Some of these virulence factors, such as the alpha toxin, which is phospholipase C, and the kappa toxin, which is a collagenase, are enzymes that hydrolyze substances essential to the integrity of membranes or other body structures . Other virulence factors, such as the beta, episolon, and iota toxins, act primarily on the vascular endothelium, causing increased capillary permeability, especially in the brain . Still others, such as the delta and theta toxins, are essentially hemolysins . Theta toxin is similar in action and serologically related to streptolysin O.

Clin Chim Acta, 1979 Mar 1, 92(2), 167 - 75
Evaluation of a rapid, sensitive and specific assay for the determination of collagenolytic activity in biological samples; Lefevere MF et al.; Several methods for the determination of collagenolytic activity were compared from the point of view of sensitivity, selectivity, simplicity and practical value for large numbers of biological samples . A labelled collagen substrate was prepared using {3H}acetic anhydride . The specificity of the assay as well as conditions allowing an optimum detection limit were investigated . The influence of low temperatures, lyophilisation and salt concentration on Clostridium histolyticum collagenase have been investigated.

Clin Orthop, 1979 Mar-Apr, (139), 92 - 6
Clostridium perfringens septic arthritis . Report of a case and review of the literature; Schiller M et al.; This is a report of Clostridium perfringens septic arthritis in a 5-year-old boy and a review of the literature on 6 previously reported cases . The treatment is the same as for other organisms producing septic arthritis.

J Biochem (Tokyo), 1979 Mar, 85(3), 833 - 8
Studies on nitrate reductase of Clostridium perfringens . II . Purification and some properties of ferredoxin; Seki S et al.; A ferredoxin was purified from Clostridium perfringens by DEAE-cellulose chromatography and Sephadex G-50 gel filtration . It had absorption maxima at 390 and 280 nm . The molecular weight was estimated to be 6,000 by Sephadex gel filtration and from the results of amino acid analysis . The isoelectric point was 3.0 . It contained four atoms of iron, four atoms of labile sulfur, and six cysteine residues . This ferredoxin as well as ferredoxin from C . pasteurianum acted as an electron donor for nitrate reductase from C . perfringens . The ferredoxin could also act as an electron donor for the hydrogenase from C . pasteurianum in hydrogen evolution.

Appl Environ Microbiol, 1979 Mar, 37(3), 496 - 504
Clostridium botulinum growth and toxin production in tomato juice containing Aspergillus gracilis; Odlaug TE et al.; The ability of spores of one type A and one type B strain of Clostridium botulinum to grow and produce toxin in tomato juice was investigated . The type A strain grew at pH 4.9, but not at pH 4.8; the type B strain grew at pH 5.1, but not at pH 5.0 . Aspergillus gracilis was inoculated along with C . botulinum spores into pH 4.2 tomato juice; in a nonhermetic unit, a pH gradient developed under the mycelial mat, resulting in C . botulinum growth and toxin production . In a hermetic unit, mold growth was reduced, and no pH gradient was detected; however, C . botulinum growth and low levels of toxin production (less than 10 50% lethal doses per ml) still occurred and were associated with the mycelial mat . The results of tests to find filterable or dialyzable growth factors were negative . It was demonstrated that for toxin production C . botulinum and the mold had to occupy the same environment.

Appl Environ Microbiol, 1979 Mar, 37(3), 433 - 7
Bacteriocin production by Clostridium acetobutylicum in an industrial fermentation process; Barber JM et al.; High titers of a noninducible bacteriocin were produced by Clostridium acetobutylicum in a molasses fermentation medium used for the industrial production of solvents . Release of the bacteriocin towards the end of the exponential growth phase was accompanied by lysis of the culture and inhibition of the production of solvents . The producer cells were sensitive to the bacteriocin, which only affected other C . acetobutylicum strains and a Clostridium felsineum strain . The thermolabile bacteriocin was not inactivated by protease enzymes and had no optimum stability between pH 4 and 5 . The sedimentation coefficient of the bacteriocin was 6S.

J Lipid Res, 1979 Mar, 20(3), 325 - 33
7alpha-Dehydroxylation of cholic acid and chenodeoxycholic acid by Clostridium leptum; Stellwag EJ et al.; The rate of 7alpha-dehydroxylation of primary bile acids was quantitatively measured radiochromatographically in anaerobically washed whole cell suspensions of Clostridium leptum . The pH optimum for the 7alpha-dehydroxylation of both cholic and chenodeoxycholic acid was 6.5-7.0 . Substrate saturation curves were observed for the 7alpha-dehydroxylation of cholic and chenodeoxycholic acid . However, cholic acid whole cell K0.5 (0.37 micron) and V (0.20 mumol hr-1mg protein-1) values differed significantly from chenodeoxycholic acid whole cell K0.5 (0.18 micron) and V (0.50 mumol-1 hr-1 mg protein-1) . 7alpha-Dehydroxylation activity was not detected using glycine and taurine-conjugated primary bile acids, ursodeoxycholic acid, cholic acid methyl ester, or hyocholic acid as substrates . Substrate competition experiments showed that cholic acid 7 alpha-dehydroxylation was reduced by increasing concentrations of chendeoxycholic acid; however, chenodeoxycholic acid 7alpha-dehydroxylation activity was unaffected by increasing concentrations of cholic acid . A 10-fold increase in cholic and 7alpha-dehydroxylation activity occurred during the transition from logarithmic to stationary phase growth whether cells were cultured in the presence or absence of sodium cholate . In the same culture, a similar increase in chenodeoxycholic acid 7alpha-dehydroxylation was detected only in cells cultured in the presence of sodium cholate . These results indicate the possible existence of two independent systems for 7alpha-dehydroxylation in C . Leptum.

Gastroenterology, 1979 Mar, 76(3), 468 - 76
Partial purification of a toxin found in hamsters with antibiotic-associated colitis . Reversible binding of the toxin by cholestyramine; Humphrey CD et al.; A toxin with cytotoxic and enterotoxic activities was isolated from cecal contents of hamsters receiving lincomycin . The toxin was partially purified by ultracentrifugation, ultrafiltration, (NH4)2SO4 precipitation, and gel filtration . Cytotoxic activity, assayed on monolayers of HeLa cells, was restricted to material that eluted in the molecular weight range of 107,000 +/- 6,000 daltons . Cytotoxicity of crude AAC toxin could be demonstrated at concentrations as low as 0.04 microgram/ml . The toxin was heat labile (55 degrees-60 degrees C for 0.5 hr) and sensitive to trypsinization, acidification at pH 3, or alkalinization at pH 9 . Cytotoxic activity was inhibited by Clostridium sordellii antitoxin . Enterotoxic activity of the crude toxin and the cytotoxic fraction from gel filtration was demonstrated by fluid secretion in ligated rabbit ileal loops . Studies were done in vitro with cholestyramine resin, vancomycin, or gentamicin to determine if the toxin was bound or denatured by these drugs . It was demonstrated that cholestyramine bound the toxin, significantly reducing its cytotoxicity . Reversible binding of the cytotoxic material was demonstrated by salt gradient elution . Neither vancomycin nor gentamicin had any effect on the in vitro cytotoxic activity of the toxin.

Lancet, 1979 Feb 3, 1(8110), 227 - 30
Prevention of necrotising enteritis in Papua New Guinea by active immunisation; Lawrence G et al.; Necrotising enteritis (pig-bel) caused by Clostridium welchii type C is a major cause of illness and death in the Highlands of Papua New Guinea . In a controlled trial of active immunisation with a clostridial toxoid prepared from type-C cultures the incidence of pig-bel in over 2500 immunised children within 24 months of immunisation was less than an eighth of that in a control group . Necrotising enteritis in Papua New Guinea is now a preventable disease.

Gastroenterology, 1979 Feb, 76(2), 356 - 61
Role of clostridial toxin in the pathogenesis of clindamycin colitis in rabbits; LaMont JT et al.; The pathophysiology of antibiotic-associated colitis was studied in rabbits with severe ileocolitis induced by oral administration of clindamycin . Cell-free, sterile filtrates of cecal contents of rabbits with clindamycin colitis contained a toxin that was lethal for mice and cytotoxic for HeLa-cell monolayers . The toxin was heat labile, was inactivated by pronase but not trypsin, and had a mol wt by gel filtration on Sephadex G-100 of 45,000 . The toxin was neutralized by antiserum to Clostridium perfringens type E, but not by other clostridial antisera . The toxin also caused severe necrosis of rabbit rectal epithelium during 18-hr organ culture, which could be completely reversed by neutralization with C . perfringens type E antiserum . These studies indicate that clindamycin colitis in rabbits is caused by overgrowth of a clostridial species, which releases a heat-labile toxic protein of mol wt of 45,000 capable of necrosing colonic epithelial cells.

J Lipid Res, 1979 Feb, 20(2), 234 - 9
12alpha-Hydroxysteroid dehydrogenase from Clostridium group P strain C48-50 ATCC No . 29733: partial purification and characterization; Macdonald IA et al.; The growth of Clostridium group P strain C48-50 {an anaerobe that contains 12alpha-hydroxysteroid dehydrogenase (12alpha-HSDH) in the absence of other dehydrogenases active upon bile salts} is greatly enhanced by the addition of 2.0% d-fructose or d-glucose to the growth medium . Other sugars were less effective . The production of NADP-dependent 12alpha-HSDH paralleled the growth of the organism which was optimal at 72 hr . Growth (and enzyme production) were suppressed by the addition of bile salt to the medium; the order of suppression was deoxycholate > chenodeoxycholate >> cholate; 1 mM of either of the dihydroxy-bile salts inhibited 96% of the growth and 100% of the enzyme production . Kinetic studies on cell-free preparations of 12alpha-HSDH revealed a pH optimum of 7.8 with greater linearity of NADP evolution with time occurring only at more alkaline pH values (9-10) . Lineweaver-Burke plots revealed Michaelis constant (K(m)) values in the range of 3-5 x 10(-4) M for deoxycholate and its glycine and taurine conjugates, while higher values were found for cholate and conjugates (K(m) value for taurocholate was 3 x 10(-3) M) . Although there was no activity with NAD, 12alpha-HSDH was shown to bind onto both NAD- and NADP-Sepharose columns, with stronger binding on the latter . The enzyme was purified 20-fold by NAD-Sepharose chromatography . The molecular weight was estimated at 100,000 by Sephadex G-200 and a series of molecular weight markers . Substrate specificity studies showed that a variety of bile salts containing 12alpha-OH groups reacted; notably, the 3alpha-sulfates of cholate and deoxycholate were nonsubstrates.-Macdonald, I . A., J . F . Jellett and D . E . Mahony . 12alpha-Hydroxysteroid dehydrogenase from Clostridium Group P strain C48-50 #29733: partial purification and characterization.

Appl Environ Microbiol, 1979 Feb, 37(2), 194 - 7
Clostridium botulinum in the Gulf of Thailand; Tanasugarn L; A survey was carried out to determine the incidence of Clostridium botulinum in samples of mud, sand, and fish from the Gulf of Thailand . Enrichment cultures from 762 samples of mud and sand from seven different areas around the Gulf were tested . C . botulinum type D was present in 10 samples, and type E was present in 2 samples taken from the west coast at Hua Hin . Enrichment cultures from 16,773 fish grouped into 2,151 samples yielded 10 filtrates containing C . botulinum type D and 5 containing type E . All of the toxic filtrates were obtained from samples of fish taken from the west coast of the Gulf of Thailand.

J Med Microbiol, 1979 Feb, 12(1), 17 - 28
The variable response of bacteria to excess ferric iron in host tissues; Miles AA et al.; The enhancement by exogenous ferric iron, both systemic and local, of the infectivity of 120 strains of bacteria, representing 17 genera, was measured in the skin of guinea-pigs . Systemic iron enhanced only 23% of 115 strains, and local iron 49% of 71 strains . Systemic iron, by an apparently anti-inflammatory action, depressed the size of lesions produced by 27 of the non-enhanced strains from nine of the genera tested . For most strains, the degree of enhancement was small, ranging from 2- to 8-fold, and often evident only with the more effective local iron; among these were some near-saprophytes like Mycobacterium phlei, M . smegmatis, Bacillus cereus and Clostridium bifermentans . Substantial enhancement, from 14- to 50-fold, was observed with the more pathogenic among the strains tested: namely BCG, Corynebacterium ovis, C . murium, Listeria monocytogenes, Erysipelothrix rhusiopathiae, Cl . perfringens, Cl . septicum, Cl . oedematiens, and some strains of Klebsiella spp., Proteus spp . and Aeromonas hydrophila . The enhancement of BCG by a single dose of iron given locally with the inoculum was only feebly manifest after 7 days, but substantial after 14--19 days, indicating the decisive effect of interference with an early humoral defence on the establishment of chronic infection some time later . Insofar as guinea-pigs whose antibacterial defences are lowered by substantial amounts of exogenous iron in the circulation represent human subjects at risk of infection because of clinical states characterised by excess of available iron, the results of the survey suggest that only a minority among the environmental bacteria can take advantage of the decreased resistance associated with such states; but that this minority is likely to include the more virulent strains in the environment.

J Clin Microbiol, 1979 Feb, 9(2), 282 - 3
Isolation of Clostridium botulinum from Honey; Midura TF et al.; Methods for the isolation of Clostridium botulinum from honey samples are described . A total of 9 of 90 honey samples were positive for C . botulinum; 6 of the positive samples had been fed to babies who developed infant botulism.

Nord Vet Med, 1979 Feb, 31(2), 81 - 6
Toxin production by Clostridium botulinum type E in fresh herring in relation to the measured oxidation potential (Eh); Huss HH et al.; Recent work has showed high positive oxidation-reduction potential (Eh) values in fresh fish flesh, whereas strongly reducing conditions exist in fish viscera and spoiled fish flesh . The present study has demonstrated that this difference in measured Eh does not significantly influence growth and toxin production by Cl . botulinum type E . In comparison, storage temperature and the spore load in fish markedly influence toxin production . The public health significance of rapid toxin formation and high toxin titers demonstrated in ungutted fish is pointed out.

J Pediatr, 1979 Feb, 94(2), 331 - 6
Honey and other environmental risk factors for infant botulism; Arnon SS et al.; Infant botulism results from the in vivo production of toxin by Clostridium botulinum after it has colonized the infant's gut . Epidemiologic and laboratory investigations of this recently recognized disease were undertaken to identify risk factors and routes by which C . botulinum spores might reach susceptible infants . Clostridium botulinum organisms, but no preformed toxin, were identified in six different honey specimens fed to three California patients with infant botulism, as well as from 10% (9/90) of honey specimens studied . By food exposure history, honey was significantly associated with type B infant botulism (P = 0.005) . In California, 29.2% (12/41) of hospitalized patients had been fed honey prior to onset of constipation; worldwide, honey exposure occurred in 34.7% (28/75) of hospitalized cases . Of all food items tested, only honey contained C . botulinum organisms . On household vacuum cleaner dust specimens and five soil specimens (three from case homes, two from control homes) contained Clostridium botulinum . The known ubiquitous distribution of C . botulinum implies that exposure to its spores is universal and that host factors contribute importantly to the pathogenesis of infant botulism . However, honey is now an identified and avoidable source of C . botulinum spores, and it therefore should not be fed to infants.

Jpn J Exp Med, 1979 Feb, 49(1), 13 - 8
Clostridium perfringens exotoxins . VI . Reactivity of perfringolysin O with thiol and disulfide compounds; Mitsui K et al.; The reactivity of perfringolysin O with thiol and disulfide compounds was studied . The activation potency of thiols was roughly proportional to the reaction rate constants of 5,5'-dithiobis-(2-nitrobenzoic acid) with thiols, which should be inversely proportional to their oxidation-reduction potentials . 1,2-Dimercaptoethane, which had the highest rate constant, most potently activated the toxin among the thiols tested and 4,4'-dipyridyl disulfide, which is known to be one of the most potent thiol-disulfide exchanging reagents, strongly inhibited toxin activity . Toxin activity was also inhibited by other thiol inhibitors.

Appl Environ Microbiol, 1979 Feb, 37(2), 181 - 6
Fluid accumulation in mouse ligated intestine inoculated with Clostridium perfringens enterotoxin; Yamamoto K et al.; Clostridium perfringens enterotoxin, when inoculated into the ligated intestinal loop of mice, caused marked distension due to fluid accumulation . The increase in weight of the intestinal loop was proportional to the log dose of enterotoxin within a range from 1 to 16 micrograms . The fluid accumulation was arrested by washing the loop with saline or by injection of the specific anti-enterotoxin serum into the loop 5 or even 30 min after inoculation of the enterotoxin . A significant increase in weight of the loop was found as early as 10 min after inoculation of the toxin . These results may suggest that entergotoxin is neither bound firmly to the mucosal membrane nor permeates into the cells of the intestinal wall . The mouse intestinal loop test is economical, simple to perform, and applicable for quantitative determination of the enteropathogenic activity of C . perfringens enterotoxin.

J Hyg (Lond), 1979 Feb, 82(1), 133 - 42
Microbiological aspects of polyphosphate injection in the processing and chill storage of poultry; Mead GC et al.; During commercial processing of broiler chickens, injection of polyphosphate (Puron 604 or 6040) resulted in microorganisms being added to the deep breast muscle . The level of contamination was related to the microbiological condition of the injection solution . Injection of polyphosphate had no effect on the shelf-life of fresh chilled carcasses held at 1 degree of 10 degrees C but changes were observed in the growth rate of microorganisms in the deep muscle and in the composition of the muscle microflora following storage . Cross-contamination of carcasses and the transfer of organisms from the skin to the deep muscle during injection was demonstrated with a marker strain of Clostridium perfringens . However, both processes were influenced by the number of marker organisms applied initially to the skin . The above findings are discussed in relation to the possible behaviour of any food poisoning bacteria present.

J Med Microbiol, 1979 Feb, 12(1), 29 - 41
Spore antigens of Clostridium sporogenes; Princewill TJ; By means of spore-agglutination of fluorescent-antibody techniques, three serological types were identified among 84 strains of Clostridium sporogenes . Four spore antigens were identified, designated A, B, C and D . A, B and C were specific for the respective types whilst D was a group antigen shared by strains of the three types . The spore antigens had corresponding somatic antigens; the type-specific somatic antigens were designated I, II, III, and the shared somatic antigen IV . The flagellar antigens were found to be type specific and were designated 1, 2 and 3; no common flagellar antigen was detected . The results of precipitation tests with spore extracts depended on the method of testing . By a capillary-tube ring method there were cross reactions among the three types of C . sporogenes, whilst by immunodiffusion in agar layers the reactions were generally type specific . In a disintegrated spore extract, two non-protein antigenic components, possibly polysaccharide, were detected by means of immunoelectrophoresis . This extract showed cross reaction with antisera to strains of all types of all types of C . sporogenes by the ring test and by immunodiffusion.

J Clin Pathol, 1979 Feb, 32(2), 148 - 52
Effect of pH on sporicidal and microbicidal activity of buffered mixtures of alcohol and sodium hypochlorite; Death JE et al.; The effect of pH on the activity of buffered sodium hypochlorite solution, and a buffered methanol/sodium hypochlorite mixture, against Bacillus subtilis spores was investigated . The best results, considering both sporicidal activity and stability, were achieved in the pH range 7.6--8.1 . The sporicidal activity and stability of five alcohol/hypochlorite mixtures, each containing a different alcohol and buffered to pH 7.6 and of hypochlorite alone buffered to pH 7.6, were compared . The mixtures were marginally more sporicidal than hypochlorite alone when fresh but were much less stable . An unbuffered methanol/hypochlorite mixture, a methanol/hypochlorite mixture buffered to pH 7.6, and hypochlorite alond buffered to pH 7.6 were all found to be effective against six vegetative organisms and spores of B . subtilis and Clostridium sporogenes . By buffering alcohol/hypochlorite mixtures or hypochlorite solution alone in the pH range 7.6--8.1, high sporicidal activity can be achieved with low concentrations of alcohol and hypochlorite . Such formulations show promise for the disinfection of heat-sensitive medical equipment.

Biochim Biophys Acta, 1979 Jan 26, 561(1), 69 - 76
The effects of temperature and ethidium bromide on the banding of heat-denatured DNA in gradients of NaI; Strayer DR; The effects of temperature and ethidium bromide on the banding of heat-denatured DNA was studied during equilibrium centrifugation in density gradients of NaI . Centrifugation at 10 degrees C prevents the partial renaturation of Escherichia coli DNA and Clostridium perfringens DNA that occurs at 20 degrees C . A centrifugation temperature of --5 degrees C is required to prevent renaturation of T7 phage DNA . Ethidium bromide decreases renaturation of Escherichia coli DNA during centrifugation at 20 degrees C and causes a small shift in the buoyant density of both denatured and native DNA . Equilibrium centrifugation at lower temperatures prevents DNA renaturation and permits increased utilization of the large buoyant density difference between native and heat-denatured DNA in gradients of NaI.

Arch Microbiol, 1979 Jan 16, 120(1), 73 - 6
Ferredoxin degradation in growing Clostridium pasteurianum during periods of iron deprivation; Schonheit P et al.; Clostridium pasteurianum was grown in batch cultures on media with an initial iron concentration of 10 micron . The uptake of iron and the synthesis of ferredoxin was followed . All the iron present in the medium was taken up by the cells before 50% of the final cell density was attained . The bacteria then continued to grow in the complete absence of exogenous iron . Ferrodoxin was synthesized during growth until the exogenous iron concentration dropped below 1 micron . During growth in the absence of iron ferredoxin was degraded with the result that at the end of growth the cells did not contain ferredoxin . The specific activity of the iron sulfur protein, pyruvate synthase (E.C . 1.2.7.1), remained constant during growth of C . pasteurianum in the absence of exogenous iron . This finding suggest that ferredoxin was used as an endogenous source of iron for the synthesis of essential iron proteins during periods of iron deprivation.

Arch Microbiol, 1979 Jan 16, 120(1), 61 - 6
Competition for L-glutamate between specialised and versatile Clostridium species; Laanbroek HJ et al.; Clostridium cochlearium could be reproducibly enriched in an L-aspartate- and L-glutamate-limited, anaerobic chemostat inoculated with anaerobic sludge . L-glutamate, L-glutamine and L-histidine were the only fermentable substrates . Less specialised clostridia of the C . tetanomorphum type could only be isolated from batch enrichments with L-glutamate and L-aspartate as energy sources . Competition experiments with C . cochlearium and C . tetanomorphum in a L-glutamate-limited chemostat resulted in the selective elimination of the latter species . Addition of glucose to the medium resulted in coexistence of both species . The molar growth yields for L-glutamate at different dilution rates at 30 degrees C were determined for both species . The maximum specific growth rates on L-glutamate were 0.55 h-1 for C . cochlearium and 0.35 h-1 for C . tetanomorphum.

J Bacteriol, 1979 Jan, 137(1), 332 - 9
Acetate assimilation pathway of Methanosarcina barkeri; Weimer PJ et al.; The pathway of acetate assimilation in Methanosarcina barkeri was determined from analysis of the position of label in alanine, aspartate, and glutamate formed in cells grown in the presence of {14C}acetate and by measurement of enzyme activities in cell extracts . The specific radioactivity of glutamate from cells grown on {1-14C}- or {2-14C}acetate was approximately twice that of aspartate . The methyl and carboxyl carbons of acetate were incorporated into aspartate and glutamate to similar extents . Degradation studies revealed that acetate was not significantly incorporated into the C1 of alanine, C1 or C4 of aspartate, or C1 of glutamate . The C5 of glutamate, however, was partially derived from the carboxyl carbon of acetate . Cell extracts were found to contain the following enzyme activities, in nanomoles per minute per milligram of protein at 37 degrees C: F420-linked pyruvate synthase, 170; citrate synthase, 0.7; aconitase, 55; oxidized nicotinamide adenine dinucleotide phosphate-linked isocitrate dehydrogenase, 75; and oxidized nicotinamide adenine dinucleotide-linked malate dehydrogenase, 76 . The results indicate that M . barkeri assimilates acetate into alanine and aspartate via pyruvate and oxaloacetate and into glutamate via citrate, isocitrate, and alpha-ketoglutarate . The data reveal differences in the metabolism of M . barkeri and Methanobacterium thermoautotrophicum and similarities in the assimilation of acetate between M . barkeri and other anaerobic bacteria, such as Clostridium kluyveri.

Am J Clin Nutr, 1979 Jan, 32(1), 244 - 50
Clindamycin-induced colitis; Fekety R et al.; The hamster model of enterocolitis after the administration of clindamycin was used to study various drugs used in treatment of the disease in humans . Current evidence strongly suggests toxigenic, clindamycin-resistant Clostridium difficile is a cause of the disease in hamster and man . This organism is susceptible to vancomycin and metronidazole, and the disease could be prevented in the hamster so long as the antibiotics were given orally . A fatal colitis almost invariably ensued once they were discontinued . Administration of cholestyramine significantly prolonged survival of hamsters, but did not pervent death or colitis . Corticosteroids or atropine-diphenoxylate (Lomotil) did not alter the disease . The hamster model may be useful in studying other kinds of treatment of this disease.

Perspect Pediatr Pathol, 1979, 5, 137 - 52
Pig Bel; Cooke RA; Pig Bel is a form of acute, segmental, necrotizing enteritis presenting as a common and life-threatening disease among the people (particularly the children) of the Highlands of Papua New Guinea . It relates to the consumption of pig meat and is thought to be caused by Clostridium welchii type C (an organism not usually present in the human intestine), the organism being transmitted to man by means of contaminated pig meat . Pib Bel resembles the diseases called "Darmbrand" which occurred in Northern Germany in the years that immediately followed World War II . Darmbrand was assocated with a Clostridium welchii infection, possibly precipitated by malnutrition . It disappeared within a few years of its recognition . Conditions that closely resemble the clinical and pathological features of Pig Bel have been reported from Uganda and Thailand . In these countries, only a few cases have been encountered and they have not been associated with the eating of pig meat or with a clostridial infection.

Z Allg Mikrobiol, 1979, 19(3), 211 - 20
Inhibition by glycine of the catabolic reduction of proline in Clostridium sticklandii: evidence on the regulation of amino acid reduction; Schwartz AC et al.; The growth of Clostridium sticklandii on the substrate pair L-alanine-L-proline (reductant-oxidant each 40 mM) in a medium containing 2 g/l yeast extract was completely inhibited by equimolar amounts of glycine, although glycine itself should be used as oxidant by the cells . The effect of glycine was the same, whether L-alanine, L-arginine, or L-serine wwere used as reductants . Performance of the growth experiments in media of high osmolarity excluded the possibility that the inhibition effected by glycine was caused by the synthesis of defective cell wall peptidoglycan . In cell-free extracts an inhibition of L-proline reduction by glycine was observed that did not belong to anyone of the known types of kinetic inhibition . It depended upon the presence of a functioning glycine-reducing enzyme system, besides glycine itself, and was lost after the purification of D-proline reductase . It was concluded from these results that a protein, besides glycine, participated in the inhibition of L-proline reduction . The regulatory implications of the inhibition for the energy metabolism of C . sticklandii are discussed.

Vet Med Nauki, 1979, 16(2), 81 - 7
{Comparative clinical, bacteriological and pathohistological studies of the sex organs of infertile cows}; Radoslavov V et al.; Investigations were performed with materials from 39 cows divided into two groups: I -- internal sexual organs of 29 cows slaughtered because of infertility; and II -- biopsy material from 10 cows that showed symptomies sterility . The same material was studied microbiologically and histopathologically by routine methods . The material was taken from the cervix, the uterus, and the horns . Bacteriologically, the presence was established of Str . uberis . E . coli, Str . dysgalactiae, Staph . et albus et citreus, Corynebacterium pyogenes, Proteus vulgaris, Str . agalactiae, Candida albicans, Clostridium sp., and others . Organisms were also isolated from the genitalia of cows showing no clinical symptoms of endometritis . Histologically, the cervix of the animals in the 1st group presented necrobiosis of the glandular epithelium, and reticulohistiocytic proliferation . Most characteristic were the changes in the uterine corpus where stasis, oedema, low necrobiotic epithelium of the glands, and a surrounding reticulocytic proliferation with the presence of lymphocytes and plasmatic cells were observed . It was found that no direct correlation between the histologic changes, the microbiologic, and the clinic results.

Arch Geschwulstforsch, 1979, 49(2), 97 - 105
{Germinating capacity of Clostridium butyricum Jena H 8 in transplantation tumors (author's transl)}; Mehnert WH et al.; Following parenteral application of spores of Clostridium butyricum their intratumoral germination and oncolysis were demonstrated on a variety of mouse transplantation tumors . The germination and propagation of the clostridia was restricted exclusively to the tumor tissue, they could not be demonstrated bacterioscopically in parallelly run normal tissue specimens . The observed differences of germination and lysis rate are due, in part, to tumor host and transplantate-specific properties.

J Wildl Dis, 1979 Jan, 15(1), 3 - 9
Antibodies to Clostridium botulinum toxins in free-living birds and mammals; Ohishi I et al.; Naturally-occuring antibodies against Clostridium botulinum toxins were found in Cathartes aura (turkey vultures), Canis latrans (coyotes) and Corvus brachyrhynchos (crows) by the passive hemagglutination (PHA) test and verified by the serum neutralization (SN) test . The prevalence of IHA antibodies was 18 of 20 vultures (90%), 5 of 12 crows (42%) and 25 to 110 coyotes (23%) . Vultures and coyotes were seropositive by the PHA test against A, B, C, D, and F toxins . The highest antibody titer 1:8192 was in vulture serum against type C . In descending order, the highest antibody levels were against type C, D, F, E, A and B toxins.

Clin Orthop, 1979 Jan-Feb, (138), 250 - 3
Clostridial myonecrosis in a patient undergoing oxacillin therapy for exacerbation of chronic foot ulcers and osteomyelitis . A case report; Miskew DB et al.; Gas gangrene developed from a chronic foot ulcer in the absence of periferal vascular disease or diabetes mellitus in a hospitalized patient undergoing parenteral antibiotic therapy . Within a 6 hour period the patient developed profound toxemia necessitating emergency and life saving leg amputation . Classically clostridial myonecrosis is diagnosed by the clinical course and the gram stain . In this case, 2 preoperative gram stains failed to show gram-positive rods . At the time of surgery, frank fasical and muscle necrosis in the peroneal compartment dictated extending the below knee amputation to above the knee . In retrospect demonstration of clostridial species and myonecrosis in the pathological specimen confirmed the clinical impression . The identified organism, Clostridium sporogenes has rarely been implicated as a cause of gas gangrene.

Trans R Soc Trop Med Hyg, 1979, 73(1), 85 - 90
Infant foods as a potential source of diarrhoeal illness in rural West Africa; Barrell RA et al.; It is common practice in rural areas of The Gambia to prepare infant foods in quantities which are sufficient to meet the needs of the day rather than one meal . These are then stored at ambient temperatures for periods up to 12 hours for feeding to the child on demand . The total viable counts and levels of Bacillus cereus, Clostridium welchii, Staphylococcus aureus and Escherichia coli were determined in 294 infant foods samples from nought to eight hours after preparation . The presence of Salmonella was determined in 10 g samples of food . In the first hour after preparation the proportion of foods dangerously contaminated was high during the rainsy season, significantly more so than during the dry season . Foods not consumed fresh were very often hazardous and almost always so after 8 hours . This problem may be a causal factor in weanling diarrhoea which also shows marked seasonal variation in prevalence.

Am J Med, 1979 Jan, 66(1), 63 - 6
Clostridium septicum bacteremia . Its clinical significance; Koransky JR et al.; The medical records of 59 patients with Clostridium septicum bacteremia were reviewed; 42 (71 per cent) of these patients had malignancies . One half had hematologic malignancies, and one half had solid tumors . Of the 21 patients with solid tumors, 14 (67 per cent) had cancer of the colon . Among these patients, the cecum was the most frequent site of malignancy . The cecum and distal ileum were the most probable portals of entry for C . septicum bacteremia among the 28 patients examined at autopsy . Patients admitted to the hospital with C . septicum bacteremia usually have fulminating clinical courses and, unless the appropriate antibiotics are administered soon after admission, the outcome is fatal . The results of this study demonstrate the high association of C . septicum bacteremia and malignancy, and the need for early recognition and therapy.

Antonie Van Leeuwenhoek, 1979, 45(1), 95 - 101
Activity of some enzymes during growth and sporulation of Clostridium botulinum type E; Emeruwa AC; The activities of alkaline and acid phosphatases, glucose dehydrogenase and NADH oxidase were assayed in cell-free extracts of sporogenic and asporogenic mutants of Clostridium botulinum . During growth of both mutants, the activities of alkaline and acid phosphatases were relatively constant, but during sporulation of the sporogenic mutant, the alkaline phosphatase activity rose to a maximum of 70 mumol/min x mg protein whereas the acid phosphatase decreased rapidly before it increased, indicating a possible role in sporogenesis . Glucose dehydrogenase activity was detected only in cell-free extracts of the sporogenic mutant and reached a maximum of 7 mumol/min x mg protein during the endospore maturation stage . The NADH oxidase activity was detected in both mutants . The NADH oxidase seems to stimulate glucose oxidation in both mutants during growth and the dehydrogenation processes of the butyric type of fermentation during spore formation in the sporogenic mutant . The findings suggest that increased glucose dehydrogenase activity in C . botulinum, as in Bacillus species, may serve as a spore event marker and that alkaline and acid phosphatases may play a regulatory role in anaerobic sporulation metabolism.

Arch Immunol Ther Exp (Warsz), 1979, 27(5), 709 - 14
Study on the immunological heterogeneity of Clostridium botulinum B type toxin; Rymkiewicz D et al.; The toxins prepared by dialysis-culture method from proteolytic (P) and nonproteolytic (NP) strains of C . botulinum were different . NP toxins from 3 strains (American, Japanese and Polish) showed a higher activation ratio, lower protein nitrogen content, and lower neutralization rate compared with P toxins . Antitoxic titers of rabbit anti-P and especially anti-NP sera were always higher when titrated with NP than with P toxins . The values of the regression coefficients in neutralization experiments were not dependent on the sera preparations, but only on the toxins . The difference between the common slope b for several sera tested against P and NP toxins was statistically significant . A difference between the passive protection of mice against P and NP toxins was observed only at very low levels of unitage . These results suggested immunological heterogeneity of B type botulinum toxin . However, for practical purposes, it is not necessary to supplement therapeutic antitoxin with a factor which neutralizes NP toxin.

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