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 w