Clostridia

Acta Pathol Microbiol Scand {B}, 1981 Oct, 89(5), 363 - 7
Enzymatic hydrolysis by bacterial phospolipases C and D of immobilized radioactive sphingomyelin and phosphatidylcholine; Malmqvist T et al.; An assay system for phospholipases C has been described with sphingomyelin immobilized to octyl-Sepharose CL-4B as substrate . The immobilization procedure was further developed and used with {14 C-choline}-sphingomyelin and {14C-choline} phosphatidylcholine (lecithin) . These immobilized radioactive phospholipids made the enzymatic assays easier to perform and made it possible to increase the sensitivity . Furthermore, since release of the choline part instead of the phosphate part of the substrate molecule was measured, it was possible to use this assay for phospholipase D as well . The enzyme characteristics of phospholipase D from Corynebacterium ovis were compared in this test system with those of three phospholipases C (from Clostridium perfringens, Bacillus cereus and Staphylococcus aureus) with respect to hydrolysing capacities and optimal ion concentrations.

Acta Pathol Microbiol Scand {B}, 1981 Oct, 89(5), 357 - 61
Enzymatic hydrolysis of immobilized sphingomyelin by three bacterial phospholipases C; Malmqvist T et al.; Through hydrophobic interaction, sphingomyelin was adsorbed to agarose beads containing octyl groups by a stepwise dilution procedure . This immobilized lipid was used as a substrate for three bacterial phospholipases C (E.C . 3.1.4.3.) . The degradation with time of this substrate showed two different fractions of the substrate according to hydrolysing velocity in the early part of the time-curve when phospholipases C from Bacillus cereus and Clostridium perfringens were used . The early fractions could be predigested by the enzymes, a procedure which resulted in linear time-curves . The corresponding early part of the time-curve for phospholipase C from Staphylococcus aureus was linear, indicating a comparatively large early fraction of the substrate for this enzyme . The stock gel of the immobilized lipid substrate could be stored for months . It was easily and reproducibly handled as a water suspension . After enzymatic hydrolysis the substrate was rapidly separated from enzyme and product by filtration . The enzyme assay presented thus represents a convenient way to avoid the difficulties connected with the use of temporary sonicated suspensions as substrate for bacterial phospholipases C.

Infect Immun, 1981 Oct, 34(1), 50 - 4
Stimulation of Clostridium perfringens enterotoxin formation by caffeine and theobromine; Labbe RG et al.; In the presence of 100 micrograms of caffeine per ml or 200 micrograms of theobromine per ml, sporulation of Clostridium perfringens NCTC 8679 rose from less than 1 to 80 or 85% . Enterotoxin concentration increased from undetectable levels to 450 micrograms/mg of cell extract protein . Heat-resistant spore levels increased from less than 1,000 to between 1 X 10(7) and 2 X 10(7)/ml . These effects were partially reversible by the addition of adenosine or thymidine . In the case of NCTC 8238, caffeine and theobromine caused a three- to fourfold increase in the percentages of cells possessing refractile spores and a similar increase in enterotoxin concentration . Heat-resistant spore levels, however, were unaffected . Inosine was ineffective in promoting sporulation in NCTC 8679.

Antimicrob Agents Chemother, 1981 Oct, 20(4), 437 - 42
Inhibition of ribonucleic acid polymerase by a bacteriocin from Bacteroides fragilis; Mossie KG et al.; The Bacteroides fragilis bacteriocin which inhibits ribonucleic acid (RNA) polymerase activity had a narrow activity spectrum in vivo and only inhibited the growth of certain B . fragilis strains . In vitro the bacteriocin was not specific and inhibited RNA polymerases from widely diverse bacterial genera . RNA polymerases from rifampin-resistant strains of Bacteroides thetaiotaomicron and Clostridium acetobutylicum were resistant to the bacteriocin in vitro . Purified bacteriocin bound to partially purified RNA polymerase, and both proteins were cosedimented in a glycerol gradient . In the RNA polymerase reaction, the bacteriocin acted as a competitive inhibitor for adenosine, cytidine, and uridine 5'-triphosphates and as a noncompetitive inhibitor for guanosine 5'-triphosphate . The bacteriocin did not inhibit RNA polymerase from chicken embryos.

Am J Gastroenterol, 1981 Oct, 76(4), 374 - 6
Pseudomembranous colitis . A complication of sulfasalazine therapy in a patient with Crohn's colitis; Pokorney BH et al.; Pseudomembranous colitis is a potentially life-threatening acute medical problem usually associated with a history of previous antibiotic exposure . Presented here is a case of sulfasalazine associated pseudomembranous colitis in a patient with known Crohn's colitis . The diagnosis was confirmed by identifying Clostridium difficile toxin in the stool . This a newly reported complication of sulfasalazine therapy.

Biochim Biophys Acta, 1981 Sep 4, 676(3), 395 - 400
Role of phosphatidylinositol in basal adenylate cyclase activity of rat heart sarcolemma; Panagia V et al.; The adenylate cyclase activity and phospholipid composition were determined in rat heart sarcolemma after treating the membranes with a phosphatidylinositol-specific phospholipase C . Complete hydrolysis of phosphatidylinositol in sarcolemma was associated with a marked loss of the basal adenylate cyclase activity . The recombination of the supernatant with the phosphatidylinositol-depleted membranes was found to reactivate the adenylate cyclase activity . The soluble component(s) in the supernatant, which restored the adenylate cyclase activity, was thermolabile and precipitated by ammonium sulfate . Extensive hydrolysis of phosphatidylcholine, phosphatidylethanolamine and sphingomyelin in sarcolemma with a Clostridium welchii phospholipase C treatment did not affect the basal adenylate cyclase activity . These results suggest that phosphatidylinositol anchors component(s) essential for the expression of basal adenylate cyclase activity to the myocardial cell membrane.

Biochemistry, 1981 Sep 1, 20(18), 5124 - 31
Transient kinetics of electron-transfer reactions of flavodoxins; Jung J et al.; Stopped-flow and laser photolysis methods have been used to investigate the rates of electron-transfer reactions of fully reduced riboflavin and the three oxidation states of Clostridium pasteurianum flavodoxin . Both normal and 7,8-dichloroflavin analogues were studied . Redox reagents included oxygen, ferricyanide, ferric EDTA, and several c-type cytochromes as oxidants and the semiquinone of 5-deazariboflavin as a reductant . The dependence of the rate of oxidation of the semiquinone form of the dichloro analogue flavodoxin upon oxidant concentration has provided clear evidence for the existence of a complex in the reaction pathway . Rate constant comparisons demonstrate that dichloro substitution decreases the rate of flavodoxin semiquinone oxidation by at least 1-2 orders of magnitude . The limiting first-order rate constants were found to be dependent on the redox potential of the oxidant, as would be predicted by theory if these were reflecting the actual electron-transfer reaction . Rate constant decreases upon chlorine substitution were also observed for the reduction of both oxidized and semiquinone forms of flavodoxin by deazariboflavin semiquinone . These results, considered in conjunction with the redox potential shift of the flavodoxin produced by the chlorine substitution, provide support for the hypothesis that electron transfer to and from the semiquinone form of the flavodoxin involves direct participation of the dimethylbenzene ring of the flavin . A comparison of oxidation rate constants for free and protein-bound fully reduced flavin suggests that the protein environment does not markedly influence coenzyme reactivity in this oxidation state.

Klin Padiatr, 1981 Sep, 193(5), 398 - 400
{Haemolysis as initial sign of Clostridium perfringens septicaemia in newborn (author's transl)}; Tollner U et al.; Cases of Clostridium perfringens, septicaemia are subject to a very gloomy prognosis in adults . This applies likewise to the rare cases reported in paediatric literature . The present report describes the fatal case of a newborn with Clostridium perfringens septicaemia . The principal sign was a severe haemolysis which failed to respond to blood exchange transfusions and antibiotic treatment . The characteristic changes of colour of the skin usually observed in septicaemia of newborn had probably been masked for some time by the reddish discolouration caused by the haemolysis, so that diagnosis was established at a late stage only.

Can J Surg, 1981 Sep, 24(5), 453 - 5
Necrotizing anaerobic infections; Bocking JK et al.; Severe necrotizing anaerobic infections carry a high mortality . From a retrospective review of 17 cases of gas gangrene, necrotizing fasciitis and nonclostridial myonecrosis seen at the Victoria and University hospitals in London, Ontario, between Jan . 1, 1975 and Dec . 31, 1977, the most common antecedent was found to be minor trauma . Location of the infection varied; the scrotum and perineum were most commonly involved . Local findings were most frequently edema, skin tenderness and induration . Mixed infections predominated with a pure growth of Clostridium sp . occurring in only three cases . The overall mortality was 29% . The mainstay of treatment after initial administration of fluids followed by broad-spectrum antibiotics is early aggressive surgical debridement of all necrotic tissue.

Am J Dis Child, 1981 Sep, 135(9), 820 - 2
Clostridium difficile associated with pseudomembranous colitis . Occurrence in a 12-week-old infant without prior antibiotic therapy; Adler SP et al.; In a previously healthy 12-week-old male infant with a two-week history of poor feeding, colic, and bloody stools, pseudomembranous colitis developed . No prior antibiotics were administered although the child had received dicyclomine hydrochloride . Clostridium difficile and its toxin were detected in the child's stool . Severe disseminated intravascular coagulopathy developed; the patient required total colectomy but eventually recovered . Clostridium difficile colonization has not, to our knowledge, been previously documented in infantile pseudomembranous colitis.

J Can Assoc Radiol, 1981 Sep, 32(3), 149 - 52
Pseudomembranous colitis in renal transplant recipients; plain film findings; Hutton L et al.; The plain abdominal film findings in nine renal transplant patients with pseudomembranous colitis were characteristic of but not specific for this entity and most commonly showed "thumb-printing" reflecting submucosal edema (five patients) and a persisting localized segmental ileus in the colon (five patients) . Based on the plain film findings, the differential diagnosis will include the acute stages of ulcerative colitis, granulomatous colitis, ischemic colitis and other inflammatory colitides, but the radiologist is in a position to suggest the correct diagnosis in the appropriate clinical setting . A barium study of the colon is not indicated, as diagnosis is usually accomplished by endoscopy . The etiology of this colitis has recently been shown to be a cytopathic toxin of the anaerobic bacterium, Clostridium difficile.

Can J Microbiol, 1981 Sep, 27(9), 942 - 8
Purification and partial characterization of proline dehydrogenase from Clostridium sporogenes; Monticello DJ et al.; A proline dehydrogenase which catalyzes the nicotinamide adenine dinucleotide (NAD) dependent oxidation of proline and the NADH-dependent reduction of delta 1-pyrroline 5-carboxylic acid (PCA) was purified from extracts of Clostridium sporogenes . Following purification, only one protein band was found on analytical polyacrylamide disc gels and on sodium dodecyl sulfate (SDS) - polyacrylamide disc gels . Sucrose density gradient centrifugation and SDS-gel electrophoresis indicated that the enzyme has a molecular weight of approximately 217 000 and consists of two subunits of equal size . During purification of proline dehydrogenase on hydroxylapatite the ratio of dehydrogenase activity to reductase activity decreases significantly, and a similar change in ratio was brought about by storage of partially purified enzyme preparations in low ionic strength buffers . Subsequent purification did not change the ratio . The dehydrogenase activity of proline dehydrogenase was inhibited by L-glutamate (Ki = 0.32 mM at pH 7.4 and Ki = 0.65 mM at ph 10.2) . However, the reductase activity of the purified enzyme was not affected by 100 mM L-glutamate.

Zh Mikrobiol Epidemiol Immunobiol, 1981 Sep, (9), 57 - 62
{Immunochemical study of Clostridium botulinum type B toxin}; Kliucheva VV et al.; Fractionation of type B . Cl . botulinum toxin, a protein complex, was carried out; as a result, 3 fractions, alpha, beta, and gamma, were isolated in a pure form, alpha-fraction, or neurotoxin, is highly toxic (5-10.10(7) LDm per 1 mg of protein), beta-fraction showed hemagglutinating activity (64-128 HAU per 1 mg of protein), gamma-fraction was not biologically active . The molecular weight of alpha and gamma-fractions was 150,000 . All these fractions had antigenic properties . alpha-fraction was serologically specific . beta- and gamma-fractions showed incomplete serologic identity.

Appl Environ Microbiol, 1981 Sep, 42(3), 477 - 83
Mechanisms of sorbate inhibition of Bacillus cereus T and Clostridium botulinum 62A spore germination; Smoot LA et al.; The mechanism by which potassium sorbate inhibits Bacillus cereus T and Clostridium botulinum 62A spore germination was investigated . Spores of B . cereus T were germinated at 35 degrees C in 0.08 M sodium-potassium phosphate buffers (pH 5.7 and 6.7) containing various germinants (L-alanine, L-alpha-NH2-n-butyric acid, and inosine) and potassium sorbate . Spores of C . botulinum 62A were germinated in the same buffers but with 10 mM L-lactic acid, 20 mM sodium bicarbonate, L-alanine or L-cysteine, and potassium sorbate . Spore germination was monitored by optical density measurements at 600 nm and phase-contrast microscopy . Inhibition of B . cereus T spore germination was observed when 3,900 micrograms of potassium sorbate per ml was added at various time intervals during the first 2 min of spore exposure to the pH 5.7 germination medium . C . botulinum 62A spore germination was inhibited when 5,200 micrograms of potassium sorbate per ml was added during the first 30 min of spore exposure to the pH 5.7 medium . Potassium sorbate inhibition of germination was reversible for both B . cereus T and C . botulinum 62A spores . Potassium sorbate inhibition of B . cereus T spore germination induced by L-alanine and L-alpha-NH2-n-butyric acid was shown to be competitive in nature . Potassium sorbate was also a competitive inhibitor of L-alanine- and L-cysteine-induced germination of C . botulinum 62A spores.

J Gen Microbiol, 1981 Sep, 126(Pt 1), 37 - 44
Extraction of spore-lytic enzyme from Clostridium perfringens spores; Gombas DE et al.; Various chemical reagents known to extract spore coat protein were used to extract spore-lytic enzyme (SLE) from intact and germinated spores of Clostridium perfringens . Of the reagents tested, 7.2 M-urea plus 10% (v/v) mercaptoethanol, pH 2.85, solubilized the most SLE activity per mg spores . The quantity of SLE extracted was dependent on the initial pH of the reagent, with a maximum between pH 2.7 and 3.0 . Germinated spores yielded more SLE than non-germinated spores upon urea/mercaptoethanol extraction . SLE release during spore germination probably utilizes a trigger mechanism not satisfied by germination alone . Significant amounts of SLE were released during germination when spores were suspended in potassium chloride or a complex germinant mixture containing brain-heart infusion, yeast extract and chloramphenicol, but not during germination with sodium nitrite, which non-enzymically lysed the cortical peptidoglycan . Greater solubilization of SLE activity was obtained by urea/mercaptoethanol extraction of spores germinated with nitrite than of spores germinated with either potassium chloride or the complex germinant.

Biull Eksp Biol Med, 1981 Sep, 92(9), 322 - 5
{Factors modulating production of antibodies against Clostridium perfringens in mammals}; Shemanova GF et al.; Immunogenic properties of Clostridium perfringens type A toxoid preparations obtained by different methods are described . As regards immunogenicity for guinea-pigs and man, toxoid obtained by the detoxification of preliminarily purified alpha-toxin (experimental toxoid) compares favourably with preparations obtained by the detoxification of alpha-toxin in a culture fluid . It was shown in experiments on guinea pigs that immunogenicity of experimental toxoid rises with the increase in the degree of purification of alpha-toxin used for detoxification . The least purified preparations turned out to be the most immunogenic for mice . In Cl . perfringens culture fluid, factor (factors) was found and separated, which reduced the immunogenicity of the respective toxoid in experiments with immunization of guinea pigs and increased it in mice . Activation of antibody-formation was disclosed to be nonspecific . The existence of this factor(s) should be taken into account in the development of vaccines and methods for evaluating their immunogenic properties.

Appl Environ Microbiol, 1981 Sep, 42(3), 394 - 9
Transformation of bile acids by Clostridium perfringens; Hirano S et al.; Thirty-five strains of Clostridium perfringens were examined for their ability to transform bile acids, both in growing cultures and by washed whole cells . All of the strains oxidized the 3 alpha-hydroxy group to an oxo group, and all except three converted the same alpha-hydroxy group into a beta-configuration . The oxidative 3 alpha-dehydrogenation was barely detectable under anaerobic cultural conditions but was clearly demonstrated in an aerated system using washed whole cells, with a pH optimum between 7.0 and 9.0 . The epimerizing reaction amounting to 10 to 20% conversion was observed in anaerobic cultures and also with resting cells, irrespective of oxygen supply . Both reactions were carried out with seven conventional 3 alpha-hydroxy bile acids, thus producing a series of 3-oxo and 3 beta-hydroxy derivatives that could be examined for gas-liquid chromatographic and mass spectrometric behavior . No evidence for the occurrence of 7 alpha- and 12 alpha-hydroxysteroid dehydrogenase activities among the test strains was found . A highly potent deconjugating hydrolase was elaborated by all of the strains.

Arch Inst Pasteur Tunis, 1981 Sep-Dec, 58(3), 355 - 73
{Thiol-dependent cytolytic bacterial toxins: streptolysin O and prominent toxins}; Alouf JE; Streptolysin O is the prototype of fifteen bacterial cytolytic protein toxins elaborated by gram-positive bacteria of species Streptococcus, Clostridium, Bacillus and Listeria . These toxins share a number of common properties: they are antigenically related as shown by cross-neutralization and immunoprecipitation; their cytolytic and other reducing agents; these toxins are inactivated by cholesterol and certain related sterols . This group of oxygen-labile cytolytic toxins has been named sulfyhdryl-activated toxins or thiol-activated cytolysins . The mechanism of action of these toxins is very likely identical or at least closely similar.

Biochim Biophys Acta, 1981 Aug 24, 665(2), 262 - 9
Bile induction of 7 alpha- and 7 beta-hydroxysteroid dehydrogenases in Clostridium absonum; MacDonald IA et al.; Eight strains of Clostridium absonum grown in the presence of 4 . 10(-4) M deoxycholate contained both NADP-dependent 7 alpha- and 7 beta-hydroxysteroid dehydrogenase activities . In one strain studied in detail, significant amounts of NADP-dependent 7 alpha- and 7 beta-hydroxysteroid dehydrogenase and NAD-dependent 7 alpha-hydroxysteroid dehydrogenase activities were demonstrated only when cells were grown in the presence of deoxycholate or chenodeoxycholate, both optimal at 4 . 10(-4) M . When the bile salt was deleted from the medium, only a trace of 7 alpha-hydroxysteroid dehydrogenase was present and 7 beta-hydroxysteroid dehydrogenase was absent . Other bile salts including cholate, ursodeoxycholate and keto bile salts were less effective as inducers . Addition of cholate to medium already containing deoxycholate at a suboptimal concentration enhanced the induction, while addition of ursodeoxycholate suppressed the induction . Further enhancement of 7 alpha- and 7 beta-hydroxysteroid dehydrogenase could be obtained by additions of deoxycholate (up to a total of 6 . 10(-4) M) during the growth of the organisms (in log phase) . As enzyme enhancement is blocked by addition of rifampicin to the medium, the authors conclude that the enzymes are bile salt-inducible . Growth curve studies revealed an optimal enzyme yield at a harvest time of approx . 6-9 h . We have preliminarily characterized several inducible enzyme components: an NADP-dependent 7 beta-hydroxysteroid dehydrogenase as well as both NAD- and NADP-dependent 7 alpha-hydroxysteroid dehydrogenases.

Biochim Biophys Acta, 1981 Aug 13, 660(2), 251 - 5
Thermostable, ammonium-activated malic enzyme of Clostridium thermocellum; Lamed R et al.; "Malic" enzyme (L-malate:NADP+ oxidoreductase (oxaloacetate-decarboxylating, EC 1.1.1.40) was purified from Clostridium thermocellum by DEAE-cellulose, agarose-NADP and Sephadex G-200 column chromatography . The 117-fold purified "malic" enzyme displayed a maximum activity of 135 units/mg at 40 degrees C and represented 0.8% of the total cell protein . Sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis analysis of the protein suggested 90% purity and an approximate tetrameric subunit molecular weight of 40 000 . The enzyme absolutely required both bivalent and monovalent cations for catalysis . Mn2+ and NH4+ were the most effective cationic activators examined . Increasing NH4+ concentration increased both enzyme activity and affinity toward L-malate . The apparent Km for L-malate was 3 X 10(-4) M at 0.4 mM NH4Cl . Enzyme activity increased linearly when temperature was raised between 22-60 degrees C and a Q10 of 2.1 was calculated from an Arrhenius plot . The enzyme was stable at heating at 60 degrees C but was denatured at higher temperatures . The enzyme half-life was 10 min at 72 degrees C . The enzyme displayed a broad pH optimum (7.2-87.2 for Tris-HCl buffer) but was inactivated by p-chloromercuribenzoate . The high thermal stability, low apparent molecular weight and NH4+ activation are properties not common to all previously described "malic" enzymes.

Biochem J, 1981 Aug 1, 197(2), 293 - 9
The release of N-acetyl- and N-glycolloyl-neuraminic acid from soluble complex carbohydrates and erythrocytes by bacterial, viral and mammalian sialidases; Corfield AP et al.; A series of substrates, sialyl(2 leads to 6)GalNAc and ganglioside GM3, containing either N-acetylneuraminic acid (AcNeu) or N-glycolloylneuraminic acid (GcNeu), has been prepared . The trisaccharide GcNeu(2 leads to 3)lactose was preapred by ozonolysis of GcNeu-GM3, and the disaccharides AcNeu(2 leads to 6)GalNAc and GcNeu(2 leads to 6)GalNAc were isolated from bovine submandibular-gland mucin by alkali elimination . Sialidases from Newcastle-disease virus, fowl-plague virus, influenza virus A2, Clostridium perfringens, Vibrio cholerae, Arthrobacter ureafaciens and human liver lysosomes were studied with the above substrates and all showed poorer cleavage of GcNeu-containing substrates when compared with the corresponding AcNeu-containing compounds . This was reflected in the Km and Vmax . values of these sialidases . Differences between viral and bacterial sialidases could be detected on the basis of their kinetic constants and time curves of sialic acid release . Preferred release of AcNeu relative to GcNeu was also observed with bovine submandibular gland mucin and a mixture of human and porcine erythrocytes, macromolecular substrates containing both AcNeu and GcNeu . The significance of differential cleavage of AcNeu and GcNeu by sialidases is considered together with examples of the role of GcNeu in physiologicaL systems.

Can J Microbiol, 1981 Aug, 27(8), 824 - 34
Stable isotope fractionation by Clostridium pasteurianum . 4 . Sulfur isotope fractionation during enzymatic S3O6(2-), S2O3(2-), and SO3(2-) reductions; Harrison GI et al.; Cell-free extracts from Clostridium pasteurianum grown on SO3(2-) utilize H2 to reduce S3O6(2-), S2O3(2-), SO3(2-) to H2S at a much faster rate than extracts from SO4(2-)-grown cells . This further supports the concept of an inducible dissimilatory type SO3(2-) reductive pathway in this organism . 35S dilution experiments further support the concept that S3O6(2-) and S2O3(2-) are pathway intermediates . The inducible SO3(2-) reductase is ferredoxin linked and the kinetics of the reduction and the sulfur isotope fractionation of the product can be altered by altering the growth conditions . The attending sulfur isotope fractionations are similar to those observed during the chemical decomposition of these compounds . In the case of S2O3(2-), 35S labelling experiments verified the conclusions derived from the stable isotope fractionation data concerning the relative reduction rates of the sulfane and sulfonate sulfurs . The reduction rates were also affected by enzyme concentration . The integrity of the whole cell is a necessary requirement for the large inverse isotope effects previously reported.

Br J Surg, 1981 Aug, 68(8), 560 - 4
The microflora of the postoperative stomach; Muscroft TJ et al.; The microflora and pH have been assessed in gastric aspirates from 163 patients after gastric surgery and have been compared with 51 patients with gastro-oesophageal carcinoma, 152 unoperated patients with peptic ulceration, 72 of whom were receiving cimetidine, 3 patients with pernicious anaemia and 27 normal subjects . The total viable bacterial count was closely related to the pH of the aspirate and was independent of the cause of the hypochlorhydria; 90 per cent of aspirates of pH 1--2 were sterile, while 94 per cent of those of pH 4--8 contained bacteria . Escherichia coli was found significantly more frequently in the postoperative and cancer groups, and Clostridium spp . were significantly more common in patients with carcinoma . Elevated gastric juice nitrite concentrations were significantly more common in aspirates of pH 4--8 and this correlated well with the presence of nitrate-reducing bacteria . The pH of an aspirate is a good indicator of the presence of bacteria and may be useful in selecting patients requiring postoperative prophylactic antibiotic cover . Hypochlorhydric patients are exposed to increased concentrations of nitrite and there may be an increased risk of gastric cancer induced by N-nitrosocompounds.

J Gen Microbiol, 1981 Aug, 125(Pt 2), 399 - 406
the involvement of Nitric Oxide in the inhibition of the phosphoroclastic system in Clostridium sporogenes by sodium nitrite; Woods LF et al.; The phosphoroclastic system was demonstrated in cell-free extracts of clostridium sporogenes by the production of carbon dioxide, acetyl phosphate, ATP and reduced NAD in the presence of pyruvate . The kinetics of acetyl phosphate production and NAD reduction were investigated . The addition of sodium nitrite to a suspension of C . sporogenes in glucose medium resulted in a rapid decrease in intracellular ATP concentration which was accompanied by an accumulation of pyruvate in the medium . This accumulation of pyruvate was caused by inhibition of phosphoroclastic system by nitrate . Nitrite inhibits this system by reaction of nitric oxide, formed from nitrate, with the non-haem iron of pyruvate:ferredoxin oxidoreductase.

Am J Vet Res, 1981 Aug, 42(8), 1302 - 5
Introduction and reisolation of selected gram-positive bacteria from fermented edible wastes; Talkington FD et al.; A fermentation process using Lactobacillus acidophilus added to edible food wastes was evaluated for its bactericidal action on selected gram-positive organisms . The Lactobacillus fermentation converts food wastes into an animal feed ingredient . In this study, 5 gram-positive bacteria of zoonotic importance were individually tested . These organisms were: Group E Streptococcus, Erysipelothrix rhusiopathiae, Clostridium perfringens, Corynebacterium pseudotuberculosis, and Listeria monocytogenes . For each experiment, Lactobacillus was first mixed into ground waste; one of the test organisms was then inoculated and mixed . This mixture was divided among eight 5.5-L containers and incubated (duplicates) at 5 C, 10 C, 20 C, and 30 C for 96 hours . Internal waste temperature, reduction-oxidation, and pH were monitored . Waste samples were taken initially and at subsequent 24-hour periods . Qualitative and quantitative recoveries of the test bacteria were attempted for each sample . Group E Streptococcus was reisolated in increasing numbers at all temperatures throughout the fermentation period . Erysipelothrix rhusiopathiae was recovered throughout the 96-hour period at 5 C; at 10 C it was recovered at 24 hours but not at 48 hours . Erysipelothrix rhusiopathiae was killed by 24 hours at 20 C and 30 C fermentation temperatures . Clostridium perfringens survived the entire test period at 5 C, 10 C, and 20 C; it was killed by 72 hours at 30 C . Neither Corynebacterium pseudotuberculosis nor Listeria monocytogenes was reisolated at any temperature at any time.

J Clin Microbiol, 1981 Aug, 14(2), 153 - 6
Hydrolytic enzymes of anaerobic bacteria isolated from human infections; Steffen EK et al.; Thirty-three strains of anaerobic bacteria isolated from human clinical specimens were examined for the presence of heparinase, hyaluronidase, chondroitin sulfatase, gelatinase, collagenase, fibrinolysin, lecithinase, and lipase activities . Pronounced heparinase activity was limited to species of the genus Bacteroides . A number of species of the genera Bacteroides and Clostridium produced hyaluronidase and chondroitin sulfatase . Gelatinase, collagenase, and fibrinolysin activities were encountered in isolates of the genera Bacteroides, Clostridium, and Peptostreptococcus . All strains capable of degrading collagen also hydrolyzed other protein substrates . Lipolytic activity was minimal among these anaerobic bacteria . No specific hydrolytic activity was consistently associated with the isolates.

J Bacteriol, 1981 Aug, 147(2), 463 - 70
Substrate specificity of citrate lyase deacetylase of Rhodopseudomonas gelatinosa and Rhodopseudomonas palustris; Giffhorn F et al.; Citrate lyase (EC 4.1.3.6) isolated from Rhodopseudomonas palustris was investigated with regard to its kinetic properties and its subunit composition . This enzyme was inactivated by citrate lyase deacetylase (EC 3.1.2.-) of Rhodopseudomonas gelatinosa . A corresponding cross-reaction was measured with partially purified deacetylase of R . palustris and citrate lyase of R . gelatinosa . The three different subunit types (alpha, beta, and gamma) of citrate lyase from R . gelatinosa wee purified to homogeneity, and antibodies were prepared against each of the three subunits and against the native enzyme complex . In corresondence with the enzymatic interactions, immunological cross-reactions were found between anti-enzyme and anti-large subunit antibodies and citrate lyase from R . palustris . On the other hand, no immunological cross-reactions were detectable among each of the antibodies and citrate lyases from Enterobacter aerogenes, Streptococcus diacetilactis, and Clostridium sphenoides . Antibodies against the large subunit of citrate lyase inhibited the deacetylase, but antibodies against the middle and small subunits did not, indicating that the large subunits of citrate lyase are involved in binding the deacetylase.

Biochim Biophys Acta, 1981 Jul 27, 654(2), 256 - 67
A basic isozyme of yeast glyceraldehyde-3-phosphate dehydrogenase with nucleic acid helix-destabilizing activity; Karpel RL et al.; A nucleic acid helix-destabilizing protein has been purified from Saccharomyces cerevisiae using affinity chromatographic techniques . Crude protein extracts at low ionic strength (approx . 0.05 M) were applied sequentially to tandem columns of native DNA-cellulose, aminophenyl-phosphoryl-UMP-agarose, poly(I . C)-agarose, poly(U)-cellulose and denatured DNA-cellulose . The 2 M NaCl eluant of the poly(U)-cellulose column was dialyzed to low ionic strength and recycled through native DNA-cellulose, poly(I . C)-agarose and poly(U)-cellulose . Purified helix-destabilizing protein eluted from the poly(U)-cellulose between 0.1 and 0.5 M NaCl . On the basis of enzymatic activity, immunological cross-reactivity, mobility on SDS gels, amino acid analysis and preliminary peptide mapping experiments, this material was identified as an isozymic fraction of glyceraldehyde-3-phosphate dehydrogenase . The major crystallizable isozyme of this enzyme from yeast is, however, considerably more acidic than the helix-destabilizing protein, and displays significantly lower helix-destabilizing activity . Stoichiometric levels of the isolated protein at low (approx . 0.01) ionic strength depress the Tm of poly(A-U) and poly {d(A-T)} by as much as 28 and 22 degrees C, respectively . Longer double helices, poly(A . U) and Clostridium perfringens DNA are also denatured by the helix-destabilizing protein, but at relatively slow rates . The binding of this protein to {3H}-poly(U) on nitrocellulose filters in {Na+}-dependent, with a 50% reduction at 0.09 M NaCl . Based on its effect on the circular dichroism spectrum of poly(A), the protein was shown to distort the conformation of the polynucleotide chain . An analogous protein from mammalian cells, P8, was also shown to depress poly(A-U) Tm.

Jpn J Antibiot, 1981 Jul, 34(7), 1078 - 81
{The absorption, excretion and influence on bowel flora of oral paromomycin sulfate (author's transl)}; Iwaki S et al.; The absorption, excretion and influence on bowel flora of oral paromomycin sulfate (aminosidine, PRM) were studied in ten normal volunteers taking a normal diet, and the following results were obtained . 1 . Serum levels of PRM were observed 0.46 micrograms/ml at a half hour, 1.14 micrograms/ml at 1 hour, 1.48 micrograms/ml at 2 hours, 0.70 micrograms/ml at 4 hours, 0.29 micrograms at 6 hours and were almost faded out at 12 hours after 4 grams of oral administration . 2 . During 0 approximately 2 hours, 2 approximately 4 hours, 4 approximately 6 hours and 6 approximately 12 hours, the mean urine concentration of PRM were observed 56.4 micrograms/ml, 56.2 micrograms/ml, 37.1 micrograms/ml and 13.8 micrograms/ml, respectively, and the total excretion in the urine by 12 hours were observed 21.14 mg (0.53%) . 3 . Oral administration of PRM caused fall in Lactobacillus, non spore-forming anaerobic Gram positive bacilli (BEP group) and Peptostreptococcus (P less than 0.001), Bacteroides (P less than 0.01) . However, after administration was discontinued, reduced bowel flora was returned to the normal range within a few days . No overgrowth of bowel flora by coliform, Clostridium or yeast was observed . Klebsiella oxytoca and toxigenic Clostridium difficile were not observed overgrowth . 4 . No side effect was observed clinically over 2 months.

Johns Hopkins Med J, 1981 Jul, 149(1), 6 - 9
Antimicrobial agents implicated in Clostridium difficile toxin-associated diarrhea of colitis; Bartlett JG; Records were reviewed for 329 patients who had antibiotic-associated diarrhea or colitis with stools showing a cytopathic toxin which is neutralized by Clostridium sordellii antitoxin . Previous studies indicate that the detection of this toxin implicate Clostridium difficile as the responsible pathogen . A spectrum of anatomical results in the colonic mucosa were found ranging from pseudomembranous colitis in 136 patients to an entirely normal endoscopic condition in 36 patients . The most frequently implicated agents were ampicillin, clindamycin, and cephalosporins . Collectively, these drugs accounted for approximately 80% of cases in which a single antimicrobial had been administered prior to the onset of symptoms . Analysis of data obtained in sequential time intervals showed a decline in the relative frequency of cases with use of clindamycin accompanied by an increase with use of cephalosporins . Less frequent drugs which were implicated in 3-8% of cases were, in rank order, penicillins other than ampicillin, erythromycin, sulfamethoxazole-trimethoprim and sulfasalazine . Tetracycline was the exclusive agent given to only three patients and there were no patients in whom chloramphenicol could be clearly implicated.

Pediatrics, 1981 Jul, 68(1), 50 - 4
Potentiation of Clostridium botulinum toxin aminoglycoside antibiotics: clinical and laboratory observations; Santos JI et al.; A child with infant botulism became apneic and died while receiving ampicillin and gentamicin therapy . As aminoglycosides have been implicated in the induction and/or exacerbation of neuromuscular blockade, we used an animal model to test the hypothesis that aminoglycosides potentiate neuromuscular blockade of botulinum toxin . In the range of aminoglycoside doses utilized in these experiments, both gentamicin and tobramycin enhance neuromuscular impairment and death of botulinum toxin-exposed mice . These results support recently published clinical observations that aminoglycosides may potentiate neuromuscular weakness caused by botulinum toxin, and suggest that these antibiotics should be used with caution in suspected cases of infant botulism.

Gastroenterology, 1981 Jul, 81(1), 5 - 9
Isolation rates and toxigenic potential of Clostridium difficile isolates from various patient populations; Viscidi R et al.; Stool specimens in various patient populations were examined to determine isolation rates of Clostridium difficile and the frequency with which this organism produces a cytopathic toxin in vitro . Clostridium difficile was isolated from 13 of 45 healthy neonates who had never received antimicrobials and the cytotoxin was detected in 12 . With 23 healthy children aged 4 to 24 mo the organism was recovered from 2 children and the cytotoxin was detected in 1 . Neither the organism nor the cytotoxin was found in specimens from healthy adults who denied antimicrobial usage during the prior 4 wk . Clostridium difficile was recovered from 12 of 56 adults receiving antimicrobials without diarrhea, but only one specimen yielded the cytotoxin . Both the cytotoxin and the organism were found in stools from most patients with antibiotic-associated colitis . Concentrations of Clostridium difficile, when present, were similar in all patient populations . Broth cultures of 165 isolates of Clostridium difficile showed all strains, but two produced the cytotoxin in vitro . However, the concentration of the toxin was substantially higher with strains recovered from patients with positive toxin assays . These observations indicate that neonates are often asymptomatic carriers of both Clostridium difficile and its cytotoxin . In adults, gastrointestinal symptoms correlated best with results of tissue culture assays and with toxigenic potential of the strains isolated.

J Biochem (Tokyo), 1981 Jul, 90(1), 295 - 8
Correspondence of the larger subunit of the MoFe-protein in clostridial nitrogenase to the nif D gene products of other N2-fixing organisms; Hase T et al.; The amino(N)-terminal sequence of the larger subunit (alpha) of the MoFe-protein from Clostridium pasteurianum was determined up to 179 amino acid residues by analyses of BrCN and tryptic peptides of the original subunit . Apparent similarities exist among the sequence of the clostridial alpha-subunit, that of the smaller subunit (beta) of the Azotobacter vinelandii MoFe protein, and those predicted from the nucleotide sequences of nif D genes in Klebsiella pneumoniae and Anabaena 7120 . In comparing the sequences of C . pasteurianum and K . pneumoniae, 45% of residues are identical of a total of 184 sites . Therefore, the larger subunit of the clostridial MoFe-protein must correspond to the nif D gene product of K . pneumoniae.

Pharmacotherapy, 1981 Jul-Aug, 1(1), 28 - 38
Metronidazole: in vitro activity, pharmacology and efficacy in anaerobic bacterial infections; Tally FP et al.; Metronidazole is a 5-nitroimidazole that has selective activity against anaerobic microorganisms, including bacteria and protozoa . Intravenous metronidazole has recently been approved by the U.S . Food and Drug Administration for the treatment of serious anaerobic bacterial infections . It is usually bactericidal at low concentrations, and its spectrum of activity encompasses almost all anaerobic bacteria and some capnophilic organisms . Anaerobic bacteria known to be resistant to metronidazole include occasional anaerobic cocci, some nonsporulating gram-positive bacilli and propionibacterium . Metronidazole is the most active antimicrobial agent against Bacteroides fragilis, the most resistant of anaerobic bacteria . Kill-curve studies demonstrate that there is a 2 to 5 log decrease in the number of colony forming units of B . fragilis and Clostridium perfringens within one hour . The only well documented metronidazole-resistant strain is a B . fragilis isolated from the normal flora of a patient on long-term metronidazole therapy for Crohn's Disease . Metronidazole resistance in Trichomonas vaginalis has recently been described in a few strains that are able to survive at increased oxygen tensions . Metronidazole has been shown to be efficacious in certain protozoal infections including trichomonal vaginitis, extraintestinal amebiasis, and giardiasis . Clinical studies have shown metronidazole to be efficacious in the therapy of a variety of anaerobic infections, including non-traumatic brain abscesses, intraabdominal sepsis, pelvic suppuration and necrotizing soft tissue infections . There have been disappointing results in the therapy of anaerobic pleuropulmonary infections with a number of superinfections caused by aerobic bacteria . Since metronidazole lacks any activity against aerobic bacteria, it must be combined with other agents, usually aminoglycosides, in the treatment of mixed infections involving anaerobic and aerobic bacteria.

J Bacteriol, 1981 Jul, 147(1), 262 - 6
Isolation of a plasmid responsible for caseinase activity in Clostridium perfringens ATCC 3626B; Blaschek HP et al.; Clostridium perfringens strain ATCC 3626B was cured of caseinase activity at a high frequency after treatment with acriflavine dye (2.5%) or elevated temperature growth (9.1%) . Caseinase-negative isolates retained the larger (9.4 megadaltons) pHB102 cryptic plasmid, but were missing the smaller (2.1 megadaltons) pHB101 plasmid present in the caseinase-positive wild-type strain . Dye-buoyant density-gradient centrifugation at 4 or 15 degrees C revealed that the pHB101 and pHB102 plasmids are temperature labile and easily converted into the nicked non-supercoiled or linear state.

Infect Immun, 1981 Jul, 33(1), 67 - 74
Intoxication of cultured human lung fibroblasts with Clostridium difficile toxin; Florin I et al.; The cytopathogenic effect of partially purified toxin from Clostridium difficile on cultured human lung fibroblasts was studied . Conditions for determination of 50% tissue culture dose were standardized . The cytopathogenic effect of the toxin was dependent on toxin concentration, exposure time, and density of the cells . Transfer of the cells to 0 degrees C did not inhibit binding of toxin to the fibroblast surface, but prevented the development of the cytopathogenic effect . Both binding of toxin and some intracellular step(s) were prevented by 2,4-dinitrophenol . These preventative effects were reversible . Before and concomitantly with the appearance of the cytopathogenic effect, the cellular uptake of uridine and of amino acids was markedly stimulated . Protein synthesis was depressed when 100% of the cells showed the cytopathogenic effect, but the synthesis of nucleic acids was inhibited only several hours later . The primary cellular target for the toxin is still unknown.

Infect Immun, 1981 Jul, 33(1), 285 - 91
Purified Clostridium difficile cytotoxin stimulates guanylate cyclase activity and inhibits adenylate cyclase activity; Vesely DL et al.; Antibiotic-associated pseudomembranous colitis has been linked with Clostridium difficile toxin . We examined the effect of toxins from four strains of C . difficile isolated from patients with pseudomembranous colitis on colonic adenylate (EC 4.6.1.1) and guanylate cyclase (EC 4.6.1.2) activities . Partially purified toxins had a cytotoxic effect on hamster fibroblasts in culture at a concentration of 10 ng/ml . Likewise, these toxins enhanced colonic guanylate cyclase activity two- to threefold, with the maximal stimulation being at 10 ng/ml . These toxins also enhanced guanylate cyclase activity in ileum, cecum, and duodenum . Both the cytotoxic activity on hamster fibroblasts and the enhancement of hamster guanylate cyclase activity were inhibited by antiserum to C . difficile toxin . These same toxins inhibited adenylate cyclase activity at a 100-ng/ml concentration, but had no effect at 10 ng/ml . They also had no effect at any concentration on colonic Na+-K+ adenosine triphosphatase . To be sure that the findings were not due to a contaminant, a purified C . difficile cytotoxin was used, and the same findings were found with the pure cytotoxin (at a 100-fold-lower concentration) . The data suggest that activation of guanylate cyclase may be a factor in the pathogenesis of antimicrobial-associated pseudomembranous colitis.

Antimicrob Agents Chemother, 1981 Jun, 19(6), 997 - 1003
Transferable tetracycline resistance in Clostridium difficile; Smith CJ et al.; The transfer of tetracycline resistance among strains of Clostridium difficile is described . Transfer occurred by a conjugation-like event that was insensitive to deoxyribonuclease, could not be mediated by donor culture filtrates or chloroform-treated donor cultures, and required cell-to-cell contact . Tetracycline-resistant progeny recovered from matings displayed a resistance phenotype identical to that of the donor in level of resistance, constitutive expression, and transmissibility . Although the original tetracycline-resistant donor contained 5 x 10(6)- and 22 x 10(6)-dalton plasmids, standard physical analyses of antibiotic-resistant transconjugants revealed no plasmid deoxyribonucleic acid molecules in common with the donor strain . Furthermore, tetracycline-susceptible derivatives of the original donor always possessed a plasmid complement identical to that of the resistant parental strain as determined by restriction endonuclease digestion analysis . The results indicate that the tetracycline resistance determinant(s) was not encoded by readily detectable plasmid deoxyribonucleic acid and may be chromosomally located.

Ann Emerg Med, 1981 Jun, 10(6), 312 - 4
Nontraumatic clostridial myonecrosis: an infectious disease emergency; Leung FW et al.; A 64-year-old man presented with a history of four days of lower abdominal pain and 12 hours of cutaneous discoloration, bullae formation, and swelling of the soft tissues of abdominal wall and right thigh . Myonecrosis of abdominal wall and an adenocarcinoma of the cecum were found at operation . Cultures of blood and fluid from the bullae yielded Clostridium septicum . Nontraumatic clostridial myonecrosis is a fulminant, usually fatal disease that is most often the result of bacteremia from an occult gastrointestinal lesion . Ulceration of the colon or terminal ileum is the most common predisposing condition, and is usually due to gastrointestinal or hematological malignancy . Patients often present with nonspecific complaints, including pain at the affected site and fever . The disease progresses rapidly to include bronze discoloration, edema, and hemorrhagic bullous lesions of the skin, subcutaneous emphysema, and myonecrosis . Presumptive diagnosis often can be made by Gram stain of the bullous fluid that reveals gram-positive bacilli and a paucity of leukocytes . Favorable outcome depends on prompt institution of appropriate antimicrobial therapy and surgical debridement of involved soft tissues, as well as correction of the underlying disorder . This disease should be considered to be a medical-surgical emergency.

Ann Emerg Med, 1981 Jun, 10(6), 307 - 11
Occult clostridial myonecrosis; Kizer KW et al.; Nontraumatic gas gangrene is extremely rare . It usually occurs following ulceration or perforation of an abdominal or pelvic viscus in persons having impaired host immune defenses . A fatal case of nontraumatic Clostridium septicum septicemia and myonecrosis in an elderly diabetic man with a perforated colon carcinoma is presented . The case illustrates the occult manner in which such cases typically present and the fulminant course of this disease . The need for rapid diagnosis and the current recommended treatment of gas gangrene are discussed.

Hoppe Seylers Z Physiol Chem, 1981 Jun, 362(6), 809 - 20
The reduction of allyl alcohols by Clostridium species is catalyzed by the combined action of alcohol dehydrogenase and enoate reductase; Bader J et al.; Cells, as well as crude extracts of Clostridium kluyveri or Clostridium spec . La 1, catalyze the hydrogenation of (E)- or (Z)-2-butenol to n-butanol . No single enzyme could be detected which directly accomplishes this reaction . It turned out that the reduction occurs as follows: 2-butenol leads to 2-butenal leads to n-butanal leads to n-butanol . The first step is catalyzed by the NAD-dependent alcohol dehydrogenase in C . kluyveri, the second by the recently detected enoate reductase which reduces not only nonactivated alpha, beta-unsaturated acylates but also alpha, beta-unsaturated aldehydes in a NADH-dependent reaction and the third step is again catalyzed by alcohol dehydrogenase . In Clostridium La 1 the alcohol dehydrogenase is NADP-dependent . The rate of the reduction of 2-butenol to n-butanol depends not only on the enzymes, but also on the ratio NAD(P)/NAD(P)H . In the presence of methylviologen cation radical which is formed by the reduction of methylviologen by the system H2/hydrogenase, the ratio NAD(P)/NAD(P)H is too small for the dehydrogenation of 2-butenol to 2-butenal . This explains the antagonistic effect of methylviologen in the hydrogenation of allyl alcohols and 2-enoates by both Clostridium species . Furthermore, the mechanism explains the finding that from a preparative point of view ethanol is a better electron donor than hydrogen for the stereospecific reduction of allyl alcohols.

Antimicrob Agents Chemother, 1981 Jun, 19(6), 980 - 6
Evaluation of eight cephalosporins in hamster colitis model; Ebright JR et al.; Eight commonly used cephalosporins were evaluated in the hamster colitis mode . They were all found to cause hemorrhagic cecitis and death within 10 days of being given as subcutaneous or oral challenges . Necropsy findings were indistinguishable from clindamycin-induced cecitis . Bacteria-free cecal filtrate obtained from hamsters dying of cephalosporin-induced cecitis contained toxin similar or identical to hat produced by Clostridium difficile isolated from the cecum of a hamster . Daily oral administration of poorly absorbed cephalosporins protected hamsters from clindamycin-induced cecitis and death as long as the cephalosporins were continued . The absorbable cephalosporins were ineffective in protecting hamsters from clindamycin-induced cecitis . This difference probably relates to the lower concentrations of absorbable cephalosporins maintained in the ceca of the hamsters . The possible correlation of these findings to human cases of cephalosporin-induced pseudomembranous colitis is discussed.

Infect Immun, 1981 Jun, 32(3), 1187 - 92
Membrane-damaging action of alveolysin from Bacillus alvei; Thelestam M et al.; We investigated membrane damage to human diploid, embryonic lung fibroblasts caused by highly purified alveolysin by measuring leakage of cytoplasmic markers and uptake of various metabolites, and we observed a leakage pattern typical of SH-activated cytolysins . However, the mode of membrane interaction resembled the mode of membrane interaction of theta-toxin from Clostridium perfringens rather than that of streptolysin O in the following respects: the activity on fibroblast membranes was high compared with the activity on sheep erythrocytes; the toxin did not bind irreversibly to fibroblast cytoplasmic membranes; considerable membrane damage was caused at 0 degrees C; and inhibition of amino acid uptake occurred in the absence of significant structural membrane damage . These findings imply that data on membrane effects caused by streptolysin O cannot be related indiscriminately to other SH-activated cytolysins . With regard to the mode of membrane interaction, two apparently different groups of SH-activated cytolysins exist.

Lab Anim Sci, 1981 Jun, 31(3), 259 - 62
Diet as a coadjuvant for development of antibiotic-associated diarrhea in hamsters (Mesocricetus auratus); Michelich VJ et al.; During a study of the effects of nutrition on experimental tumors in hamsters, fatal diarrhea developed . To determine the role of diet in this condition and the relationship between diet and antibiotics, two diets were used, ground commercial diet and a purified diet . Two antibiotics were used, neomycin sulfate and vancomycin . Diarrhea was evident soon after the animals were given the combination of purified diet and neomycin sulfate . Vancomycin initially acted as a suppressor of diarrhea, but hamsters fed the purified diet developed diarrhea after the drug was discontinued . None of the animals fed ground commercial diet treated with neomycin sulfate developed diarrhea . Clostridium difficile was isolated from most of the sick animals; all of the isolates of Clostridium difficile were toxigenic except one . No significant differences were seen among the Clostridium difficile isolated from different animals . Clostridium sporogenes and Clostridium tertium also were isolated from some of the animals.

Biken J, 1981 Jun, 24(1-2), 13 - 21
Complementation characteristics of newly isolated mutants from two groups of strains of Clostridium perfringens; Imagawa T et al.; 1 . Mutants of a group were converted to b group by a second NG treatment . The resulting b group mutants could not produce the marker products that had been lost on the first NG treatment but could produce the others by complementation . 2 . Mutants of b group were converted to constitutive mutants by a second NG treatment . No back mutation from b group was observed.

Biken J, 1981 Jun, 24(1-2), 1 - 11
Further studies on complementation between mutants of Clostridium perfringens; Tatsuki T et al.; 1 . Mutants devoid of lambda- and kappa-toxin and hemagglutinin (HA), respectively, were isolated from Cl . perfringens PB6K . The lambda- and HA- mutants could be classified into a and b groups by complementation but the kappa- mutants were all of the a group . 2 . All b group mutants isolated, irrespective of the marker used for isolation, were pleiotropically negative or leaky with respect to theta-, lambda- and kappa-toxin and HA production . 3 . Lambda-toxin produced by complementation was proved to be a rennet-like protease . 4 . The activities of 12 extracellular enzymes, including sialidase, of several b group strains and the parent PB6K were compared, but no definite differences were observed . From this finding, the productions of these enzymes were concluded not to be regulated by the same mechanism as theta-, lambda- and kappa-toxin and HA . 5 . Cl . perfringens CN3870 was also studied . Findings were similar to those on PB6K except for very low activity of HA.

Ann Emerg Med, 1981 Jun, 10(6), 315 - 7
Clostridium perfringens: an unusual case of septic arthritis; Harrington TM et al.; Anaerobic joint infections may be a more common occurrence than previously reported . A nine-year-old with a septic knee due to Clostridium perfringens following a puncture wound was seen in the emergency department . Diagnosis was established by isolating the organism from an anaerobic synovial fluid culture . The patient was treated successfully with open drainage and parenteral penicillin . This case illustrates the necessity for specifically requesting anaerobic cultures on synovial fluid of patients with acute septic monoarthritis.

Biochim Biophys Acta, 1981 May 29, 668(3), 325 - 32
Trypsin activation of enterotoxin from Clostridium perfringens type A: fragmentation and some physicochemical properties; Granum PE et al.; Clostridium perfringens type A enterotoxin was activated about 3-fold by treatment with trypsin, without an observed change in molecular weight . On denaturation in 8 M urea, the trypsinated enterotoxin lost a small peptide of about 4000 daltons . The single cysteine residue of enterotoxin was in the small peptide together with seven out of nine residues of proline . Trypsin activation, without removal of the small peptide, increased the 'outside' number of amino groups from eight to eleven . The trypsin treatment of the enterotoxin did not change the antigenic properties of the protein . Glycine was the C-terminal residue of the native enterotoxin while the dansyl alpha-amino acid of the N-terminal could not be identified.

J Biol Chem, 1981 May 25, 256(10), 5153 - 61
RNA polymerase from Clostridium acidi-urici . Characterization of a naturally occurring rifampicin-resistant bacterial enzyme; Murray CL et al.; We report here the isolation of a prokaryotic RNA polymerase that shows pronounced template specificity . The enzyme from Clostridium acidi-urici is highly active on DNA templates isolated from phage that infect Gram-positive organisms and is essentially inactive at either high or low ionic strength on DNA from phage associated with Gram-negative bacteria . The enzyme is also unique among RNA polymerases isolated from wild type bacteria in being highly resistant to inhibition by rifampicin . These properties are characteristic of the enzyme present in several independently isolated strains of C . acidi-urici . However, RNA polymerase present in other clostridial species resembles the enzyme present in Bacillus subtilis in sensitivity to rifampicin and template specificity.

Wien Klin Wochenschr, 1981 May 15, 93(10), 331 - 6
{Anaerobic bacteria in clinical specimens of a hospital laboratory (author's transl)}; Mittermayer H; Over a 14-month period 261 clinical specimens were cultured anaerobically, this figure representing about 3% of the total specimens submitted to the microbiological laboratory . Not accepted for anaerobic culture were stools, sputa, urines, vaginal and urethral swabs and swabs from superficial skin lesions . All materials were sent to the laboratory in an anaerobic transport medium and processed according to the recommendations of the American Society for Microbiology and the Center for Disease Control, Atlanta, Georgia . Not included in this figure are blood cultures and cultures from removed venous catheters . 35% of the specimens contained anaerobes . 169 anaerobic strains were isolated . 78 of them belonged to the Bacteroides fragilis group . Clostridium perfringens was found in only 7 cases . 14% of the strains were B . melaninogenicus, 7% other gram-negative non-sporeforming rods . In most of the cases a mixed aerobic-anaerobic flora was found . More than 75% of specimens obtained from infections originating from the lower intestinal tract yielded growth of anaerobes . In infections of the bile duct anaerobes were found in 25% of the cases . In infections of the upper urinary tract anaerobes played no major role . 35% of the isolates of the B . fragilis group were resistant to tetracycline, 17% to cefoxitin and 9% to clindamycin.

Experientia, 1981 May 15, 37(5), 451 - 2
On the production of 12 alpha-hydroxysteroid dehydrogenase from Clostridium group P, strain C48-50 ATCC 29733; Macdonald IA; The production of 12 alpha-hydroxysteroid dehydrogenase of Clostridium group P strain C48-50 was optimized when the organism was grown in the presence of 2% fructose and 0.1% dithiothreitol . It appears that an initial redox potential of less than -160 mV (achieved by autoclaving in the presence of dithiothreitol, dithioerythritol or cysteine) is important in the production of this enzyme.

Biochemistry, 1981 May 12, 20(10), 2908 - 16
Phase behavior of ether lipids from Clostridium butyricum; Goldfine H et al.; Ether lipids have been isolated from the phospholipid fraction of Clostridium butyricum IFO 3852 cells which had been grown in media devoid of biotin with added elaidic acid or oleic acid . The plasmalogen form of phosphatidylethanolamine (plasmenylethanolamine) from elaidate-grown cells was highly enriched with 18:1 in both the alk-1-enyl and acyl chains . A transition from the gel to liquid-crystalline state, with a peak maximum (Tm) at 33 degrees C and enthalpy delta H = 5.7 kcal/mol, was observed by differential scanning calorimetry . With the fluorescent probes cis- and trans-parinaric acids, transitions were observed at 33 degrees C on heating and at 29 degrees C on cooling . These transition temperatures are 5-6 degrees C lower than those reported for the corresponding diacyl lipid, dielaidoylphosphatidylethanolamine . A similar study of the phase behavior of both the elaidate-enriched and oleate-enriched glycerol acetal derivative of plasmenylethanolamine from C . butyricum revealed a large hysteresis of 12.5-16 degrees C . Hysteresis in the polar head group motion was also observed by 31P nuclear magnetic resonance . The elaidate-enriched lipid, which melted between 28 and 33 degrees C, appears to undergo supercooling prior to the transition to the gel state at about 18-13 degrees C, depending on the scanning rate . The formation of a more ordered gel state relative to plasmenylethanolamine was indicated by a 2-fold increase in delta H . Electron microscopy revealed a marked reorganization from typical multilamellar liposomes above Tm to large needle-like structures below Tm . The oleate-enriched glycerol acetal lipid formed the gel phase at -4 degrees C, which is 10 degrees C above the transition temperature reported for dioleoylphosphatidylethanolamine . Stabilization of oleate-enriched glycerol acetal lipid bilayers may result from hydrogen bonding between polar head groups . The relationship of the phase behavior of the ether lipids to the lipid composition and phase behavior of C . butyricum membranes is discussed.

Mikrobiologiia, 1981 May-Jun, 50(3), 458 - 66
{Fermentation products and the pectolytic enzymatic activity in Clostridium felsineum strains differing in intensity of sporulation}; Avrova NP et al.; Clostridium felsineum strains differing in the activity of pectolytic enzymes and the rate of spore formation varied also in the synthesis of fermentation products . Strains with the high rate of spore formation synthesized less butyric and propionic acids, but twice as much butanol; as a result, the proportion between butyric acid and butanol was 8 times as low as in the strains forming a small quantity of spores . The strains with the high activity of pectolytic enzymes contained 6.7 times more acetic acid, 3.5 times more ethanol and 2.7 times less propanol.

J Am Acad Dermatol, 1981 May, 4(5), 619 - 29
Pseudomembranous (antibiotic-associated) colitis; Saco LS et al.; We have come to understand the cause of antibiotic-associated pseudomembranous colitis (PMC) only in the last decade . Clostridium difficile produces the intestinal dysfunction and the characteristic finding of exudative plaques on the mucosa by elaborating a toxin in the colon . This report reviews the development of our knowledge of this disease and the rapid adoption of a rational therapy once the cause was specified . C . difficile or its toxin can be cultured or isolated from the stools of 90% of the patients with PMC . This organism is almost never found in healthy people or in any other conditions except inflammatory bowel disease, where its significance is not yet known . The detection of pseudomembranes by sigmoidoscopy establishes the diagnosis . The laboratory technics that confirm the presence of C . difficile and its toxin are being incorporated into many laboratories around the country . Treatment of diagnosed PMC is relatively simple and usually completely effective . The offending antibiotic is stopped, a proper fluid and electrolyte balance maintained, and oral vancomycin begun, 125 to 500 mg four times a day . Cholestyramine can also be used as an adjunct to this regimen . Relapse can occur in patients treated with oral vancomycin, necessitating a repeat course of therapy.

Appl Environ Microbiol, 1981 May, 41(5), 1184 - 91
Relationship of sporulation, enterotoxin formation, and spoilage during growth of Clostridium perfringens type A in cooked chicken; Craven SE et al.; Sporulation and enterotoxin formation were determined for 17 strains of Clostridium perfringens type A in autoclaved chicken dark meat and in Duncan-Strong sporulation medium . The mean numbers of heat-resistant spores detected after 24 h at 37 degrees C were log10 1.13 to log10 7.64/ml in Duncan-Strong medium and log10 4.93 to log10 6.59/g in chicken . Of 17 strains, 7 formed enterotoxin in Duncan-Strong culture supernatant (1.0 to 60 microgram/ml) and 8 produced enterotoxin in chicken (0.21 to 24 microgram/g) . Additional studies with chicken were conducted with C . perfringens NCTC 8239 . With an inoculum of 10(6) cells per g, greater than log10 7.99 vegetative cells per g were detected by 4 h in chicken at 37 degrees C . Heat-resistant spores occurred by 4 and 6 h and enterotoxin occurred by 8 and 6 h in autoclaved chicken dark meat and barbecued chicken drumsticks, respectively . Enterotoxin was detected in autoclaved dark meat after incubation at 45 degrees C for 1.5 h followed by 37 degrees C for 4.5 h, but not after incubation at 45 degrees C for 1.5 to 8 h . With an inoculum of 10(2) cells per g in oven-cooked or autoclaved chicken, greater than log10 8.00 vegetative cells per g were detected by 6 to 8 h at 37 degrees C, heat-resistant spores were detected by 8 h, and enterotoxin was detected by 12 h . A statistical analysis of odor determinants of chicken after growth of C . perfringens indicated that, at the 95% confidence level, the product was considered spoiled (off or unwholesome odor) by the time spores or enterotoxin were formed.

J Clin Microbiol, 1981 May, 13(5), 940 - 6
Highly sensitive assay for Clostridium perfringens enterotoxin that uses inhibition of plating efficiency of Vero cells grown in culture; McDonel JL et al.; A highly sensitive and reproducible biological assay for Clostridium perfringens enterotoxin is described that uses Vero (African green monkey kidney) cells grown in tissue cultures . Very small doses of the enterotoxin inhibited the plating efficiency of the cells . This inhibition of plating efficiency could be used to detect as little as 0.1 ng (1 ng/ml) of enterotoxin, and a linear dose-response curve was obtained with 0.5 to 5 ng (5 to 50 ng/ml) . A nonlinear, but reproducible, curve was obtained with a dose range from 0.1 to 100 ng (1 to 1,000 ng/ml) . A new unit of biological activity, called the plating efficiency unit, was defined as that amount of enterotoxin that caused a 25% inhibition of the plating of 200 cells inoculated into 100 microliters of medium in a microwell culture system . One milligram of highly purified enterotoxin contained about 400,000 plating efficiency units . Additional studies demonstrated that the biological and serological activities of the enterotoxin molecule were not equally labile.

Am J Physiol, 1981 May, 240(5), C234 - 8
Enzymatic isolation of cells from bone: cytotoxic enzymes of bacterial collagenase; Hefley T et al.; The enzymatic isolation of cells from fetal rat calvaria is most effectively achieved with crude Clostridium histolyticum collagenase . However, this bacterial collagenase damages the cells during the digestion of the tissue . We have used cell density, as measured by isopycnic centrifugation on polysucrose gradients, as an indicator of cell damage . There are at least two enzymes in crude bacterial collagenase capable of damaging the cells in this tissue . One of these is clostripain that has been well characterized . The other cytotoxic enzyme is uncharacterized, and its effects are not evident until the clostripain activity has been inhibited by alpha-tosyl-lysyl chloromethane . The apparent activity of this second enzyme can be inhibited by withholding magnesium from the digestion medium and by increasing the potassium concentration of the digestion medium.

Arch Surg, 1981 May, 116(5), 537 - 40
Elective cholecystectomy . The role of biliary bacteriology and administration of antibiotics; Farnell MB et al.; One hundred patients who underwent elective cholecystectomy were studied prospectively to define the accuracy of an intraoperative Gram's stain procedure in discovering the presence of bactibilia, the apparent necessity for prophylactic use of antibiotics, and the resulting septic complication rate . Antibiotics were used for prophylaxis in 48 patients and were withheld from 52 . Intraoperative Gram's stain procedure and culture were used in all cases . Bactibilia was present in 23% of the group and was accurately predicted by Gram's stain in 87% of all . The only septic complication was a stitch abscess in one patient in the group that received antibiotics . The presence of bactibilia does not seem to be associated with an increased incidence of infectious complications in elective cholecystectomy for chronic cholecystitis . Routine use of Gram's stain should detect the presence of Gram-positive rods (Clostridium), thus allowing early initiation of appropriate treatment.

J Biol Chem, 1981 Apr 25, 256(8), 3712 - 7
Separation and characterization of heavy and light chains from Clostridium botulinum type C toxin and their reconstitution; Syuto B et al.; Clostridium botulinum type C toxin consists of a heavy and a light chain with molecular weights of 98,000 and 53,000, respectively, which are linked by one disulfide bond . The two components were separated from each other by quaternary aminoethyl Sephadex A-50 column chromatography by stepwise elution with NaCl in 27.5 mM borax-45 mM sodium dihydrogen phosphate buffer, pH 8.0, containing 5% 2-mercaptoethanol at 0 degrees C . The purified components had different amino acid compositions and antigenicities, and the toxicity of the toxin was neutralized completely by either anti-heavy chain Fab or anti-light chain Fab . the two components could be reconstituted to form an active molecule with recovered toxicity which varied according to the method used . Maximum recovery was obtained in a system in which the intersubunit S--S bond was first formed in the presence of high concentration of neutral salts, after which the concentration of salt was gradually decreased . The reconstituted preparation was highly toxic and had the same properties as the parental toxin on chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and immunodiffusion . By the use of three perturbants, the fractions of exposed tryptophans and tyrosines of the preparation were found to be almost the same as that of the parental toxin.

Biochim Biophys Acta, 1981 Apr 17, 674(1), 118 - 27
Influence of cations on lysozyme-induced germination of coatless spores of Clostridium perfringens 8-6; Sacks LE; Bacterial endospore germination is powerfully influenced by inorganic salts, cations having especially important effects . Spores of Clostridium perfringens 8-6 are unusual in lacking a spore coat; these spores germinate only in the presence of lysozyme, which readily digests the exposed cortex . Lysozyme-induced germination showed the same response to ionic strength and valence of cations as does lysozyme hydrolysis of peptidoglycan, and close parallels are evident in the influence of inorganic cations on germination of normal spores . La3+ and transition element cations inhibited lysozyme-induced germination at low concentration, again demonstrating parallels with their action on lysozyme digestion of peptidoglycan and on the germination of normal spores . The poly-cations poly(L-lysine) and Ruthenium Red inhibited at extremely low concentrations . Mn2+ and Co2+, at appropriately low concentrations, stimulated lysozyme germination of 8-6 spores and also lysis of Micrococcus lysodeikticus.

Jpn J Med Sci Biol, 1981 Apr, 34(2), 61 - 8
Separation of Clostridium botulinum type A derivative toxin into two fragments; Kozaki S et al.; Two fragments with molecular weights of 105,000 (fragment I) and 58,000 (fragment II) were separated chromatographically from each other after Clostridium botulinum type A derivative toxin adsorbed onto a QAE-Sephadex column was treated with dithiothreitol and urea . They were antigenic and formed crossing precipitin lines against anti-derivative toxin in agar gel diffusion tests . Upon removal of dithiothreitol and urea by dialysis, the two fragments reassembled to reconstruct the derivative toxin molecule.

J Clin Pathol, 1981 Apr, 34(4), 416 - 9
IgG antibodies against anaerobic bacteria in blood donor sera; Schwan A; Fifty-six blood donor sera were examined by indirect immunofluorescence for IgG antibodies to a selection of anaerobic bacterial strains, often isolated from wound cultures . Another 25 sera were examined by IgG antibodies to six NCTC Bacteroides strains . A wide range of IgG antibody titres were found against Fusobacterium, Clostridium, and anaerobic streptococcal species . Very low titres were found against the Bacteroides strains . It is suggested that the testing of a single serum specimen for IgG antibodies against a representative collection of Bacteroides fragilis strains could be helpful in verifying a clinical suspicion of infection with these organisms.

J Clin Pathol, 1981 Apr, 34(4), 414 - 5
Antitoxin production in antibiotic-associated colitis?
Lishman AH, Al-Jumaili IJ, Record CO.
The production of antitoxin after Clostridium difficile-induced diarrhoea has not been reported previously . The stool of a patient with prolonged antibiotic-associated diarrhoea contained C . difficile toxin, and the serum neutralised the cytopathic effect of C . difficile toxin in tissue culture.

Appl Environ Microbiol, 1981 Apr, 41(4), 1074 - 7
Identification, distribution, and toxigenicity of obligate anaerobes in polluted waters; Daily OP et al.; A seasonal occurrence of obligately anaerobic bacteria, predominantly of the genera Bacteroides and Clostridium, in a polluted water site has been observed . The number of anaerobes varied from 1.8 X 10(3) cells/ml in the warmer months to 10 cells/ml in winter . Several isolates were toxigenic, indicating a potential human health hazard.

Arch Intern Med, 1981 Apr, 141(5), 663 - 4
Recurrent pseudomembranous colitis unassociated with prior antibiotic therapy; Moskovitz M et al.; Fulminant and recurring pseudomembranous colitis developed in an elderly woman without prior antibiotic administration within the previous year . Stool culture yielded Clostridium difficile, and tissue cultures showed C difficile cytotoxin . Treatment with vancomycin hydrochloride was initially successful, but a serious relapse after its discontinuation necessitated low-dose long-term prophylaxis . Clostridium difficile may be responsible for pseudomembranous colitis unassociated with antibiotic therapy.

Am J Med, 1981 Apr, 70(4), 906 - 8
Epidemiology of antibiotic-associated colitis; isolation of Clostridium difficile from the hospital environment; Fekety R et al.; Clostridium difficile is the most important cause of antibiotic-associated colitis . Using selective media, it was found that contamination with this organism was common in the environment of patients in the hospital with the disease . It was often found on floors, hoppers, toilets, bedding, mops, scales and furniture . This organism was also present on these items, but less often, in areas in which patients known to carry this hardy spore-forming organism had not been detected . Air, food and walls were negative . The organism was isolated from the hands and stools of asymptomatic hospital personnel . It was also found on surfaces in a patient's home . The importance of the various sources of the organism in its spread in the hospital is not known, and further studies are needed . It is suggested that enteric isolation precautions, and careful handwashing and cleansing of potentially contaminated surfaces and objects may be worthwhile when cases of antibiotic-associated colitis are identified.

J Am Vet Med Assoc, 1981 Apr 1, 178(7), 732 - 4
Suppurative abomasitis associated with Clostridium septicum infection; Eustis SL et al.; A 1-week-old calf and two 1-month-old lambs from different farms in eastern South Dakota had severe abomasitis characterized by diffuse suppurative inflammation, edema, and emphysema . Clostridium septicum in the abomasal wall of each was identified by immunofluorescence and isolation . The lesions and microbiologic findings established the disease as braxy, a condition infrequently reported in the United States and thought to affect sheep only.

Gastroenterology, 1981 Apr, 80(4), 697 - 70
Occurrence of Clostridium difficile toxin during the course of inflammatory bowel disease; Meyers S et al.; Clostridium difficile toxin, the presumed mechanism of antibiotic-associated pseudomembranous colitis, has been suggested as a contributory factor to mucosal injury in inflammatory bowel disease . We evaluated its incidence and apparent role in 65 consecutive patients with diarrheal and inflammatory bowel diseases . Toxin was demonstrated in 3 of 18 patients with ulcerative colitis (17%), 1 of 26 with Crohn's colitis (4%), and 5 of 21 with a variety of diarrheal illnesses (24%) . Toxin appeared only in those who had been exposed to antimicrobials within 2 mo . In inflammatory bowel disease, presence of toxin was not correlated with disease severity . We conclude that Clostridium difficile toxin appears only in patients exposed to antimicrobials and is unlikely to be a significant contributory factor in inflammatory bowel disease.

Appl Environ Microbiol, 1981 Apr, 41(4), 1078 - 80
Microbial profiles of country-cured hams aged in stockinettes, barrier bags, and paraffin wax; Draughon FA et al.; No significant differences were found in surface microflora of county-cured hams covered with stockinettes, barrier bags, or a coating of paraffin wax during aging, except for a reduction in mold growth on waxed hams . The incidence of Clostridium spp . was lost in all treatments . Micrococcus spp . and Streptococcus spp . were the most common contaminants, but caused no apparent spoilage problem in any treatment.

Am J Epidemiol, 1981 Apr, 113(4), 436 - 44
Restaurant-associated type A botulism: transmission by potato salad; Seals JE et al.; In the period November 13-18, 1978, seven cases of type A botulism occurred in persons who had eaten in a restaurant in Colorado . The outbreak was recognized when two persons who had independently eaten at the restaurant were hospitalized with an illness compatible with botulism . Surveillance efforts identified five additional cases . Potato salad made at the restaurant and available for service during an 11-day period was epidemiologically incriminated as the vehicle of botulinal toxin transmission (p less than 0.00001) . Laboratory studies showed that Clostridium botulinum spores on the surface of potatoes could survive baking in the manner used by the restaurant and that botulinal toxin could be produced in potatoes contaminated with C . botulinum spores.

J Neurosurg, 1981 Apr, 54(4), 455 - 60
High-dose BCNU with autologous bone marrow rescue for recurrent glioblastoma multiforme; Hochberg FH et al.; Eleven patients with recurrent malignant glioma were treated with single high doses of BCNU ranging from 600 to 1400 mg/sq m . To prevent the characteristic late myelosuppression observed after conventional doses of BCNU, autologous bone marrow harvested just before drug treatment was infused 24 to 36 hours after therapy . Higher doses of BCNU causes earlier and more profound myelosuppression; one patient died on pancytopenia, breakdown of the gut epithelium, and Clostridium septicemia 10 days after receiving 1400 mg/sq m of BCNU . All patients experienced transient emesis; four developed transient elevation of hepatic enzymes, two reversible interstitial pulmonary infiltrates, and two who received 1400 mg/sq m BCNU suffered irreversible cortical damage . Eight patients receiving 600 to 1200 mg/sq m demonstrated reconstitution of polymorphonuclear leukocytes an platelets within at least 30 days after treatment . With a follow-up time of up to 19 months, four patients improved, three stabilized, and three deteriorated and died . The median survival time was 7 months . Computerized tomography performed on patients receiving constant corticosteroids showed diminished contrast enhancement and mass effect in eight patients . High-dose BCNU at doses up to 1200 mg/sq m with marrow rescue is a feasible approach to the treatment of patients with glioblastoma.

J Biol Chem, 1981 Mar 25, 256(6), 2808 - 12
Nucleotide sequence of the gene coding for the nitrogenase iron protein from Klebsiella pneumoniae; Sundaresan V et al.; We report the complete DNA sequence of the Klebsiella pneumoniae nifH gene, the gene which codes for component 2 (Fe protein or nitrogenase reductase) of the nitrogenase enzyme complex . The amino acid sequence of the K . pneumoniae nitrogenase Fe protein is deduced from the DNA sequence . The K . pneumoniae Fe protein contains 292 amino acids, has a Mr = 31,753, and contains 9 cysteine residues . We compare the amino acid sequence of the K . pneumoniae protein with available amino acid sequence data on nitrogenase Fe proteins from two other species, Clostridium pasteurianum and Azotobacter vinelandii . The C . pasteurianum Fe protein, for which the complete sequence is known, shows 67% homology with the K . pneumoniae Fe protein . Extensive regions of strong conservation (90-95%) are found, while other regions show relatively poor conservation (30-35%) . It is suggested that these strongly conserved regions are of special importance to the function of this enzyme, and the findings are discussed in the light of evolutionary theories on the origin of nif genes.

J Biol Chem, 1981 Mar 10, 256(5), 2199 - 206
Magnetic interactions between dysprosium complexes and two soluble iron-sulfur proteins; Blum H et al.; A tetranuclear ferredoxin from Rhodopseudomonas gelatinosa {4Fe-4S}(+2,+3) has been examined by electron paramagnetic resonance techniques . The temperature-dependence and relaxation characteristics of the spectral lines indicate that the spin-lattice interaction is described at low temperatures by a T2 law which gives way to a T9 Raman relaxation as the temperature is raised . At higher temperatures an Orbach process becomes dominant . In the presence of dysprosium complexes the relaxation and line widths are modified . From crystallographic structure determinations of similar proteins we are able to relate the dysprosium effects to the spatial separation between the complex and the tetranuclear cluster . This scale is then tested against a ferredoxin from Clostridium pasteurianum which contains two tetranuclear clusters, {4Fe-4S}(+1,+2) . We find that for these soluble iron-sulfur proteins the dysprosium complexes form a shell at the protein surface . The magnetic interaction between the clusters and the complexes altering the relaxation time goes as r-6, while the low temperature line broadening is described by an r-3 dipole interaction.

Schweiz Med Wochenschr, 1981 Mar 7, 111(10), 350 - 5
{Mild spontaneous course of a case of pseudomembranous colitis . Case report and literature review}; Jaeger A et al.; Pseudomembranous colitis (PMC) caused by a toxin produced by Clostridium difficile is described in the literature as a severe diarrheal disease with a high mortality rate . A case which tends to absolve PMC from this reputation is reported involving an outpatient who developed well documented PMC subsequent to ampicillin therapy but required no treatment . The number of unreported cases of antibiotic-associated colitis with and without pseudomembrane formation is probably very high, since only severe cases of diarrhea are thoroughly investigated . In a chronological literature review an attempt is made to update the nomenclature of antibiotic-associated colitis . There are recent reports of a connection between the Clostridium difficile toxin and the chronic inflammatory "non-bacterial" intestinal diseases ulcerative colitis and Crohn's disease . The authors finally consider whether in cases of antibiotic-associated diarrhea efforts should be made to isolate Clostridium difficile and/or demonstrate the presence of its toxin, for the purposes of prognosis and therapy.

Biochim Biophys Acta, 1981 Mar 6, 641(2), 401 - 9
Protective effects of osmotic stabilizers on morphological and permeability alterations induced in Vero cells by Clostridium perfringens enterotoxin; McClane BA et al.; Culture medium made hypertonic by the addition of osmotic stabilizers such as sucrose, poly(ethylene glycol), dextran and bovine serum albumin protected against changes in morphology and plasma membrane permeability induced by Clostridium perfringes enterotoxin . The protection did not appear to be due to binding inhibition . Results of these studies support an osmotic disruption mechanism for the action of the enterotoxin . A comprehensive model of the enterotoxin's action based on an osmotic disruption mechanism is proposed.

Zentralbl Bakteriol A, 1981 Mar, 249(1), 99 - 103
Oxygen-dependent metronidazole-resistance of Clostridium histolyticum; Fuzi M; Sensitivity determinations with 6 strains of Clostridium histolyticum showed that the inhibitory action of metronidazole was highly dependent on the oxygen concentration of the environment . Under anaerobic conditions they were sensitive but at increased oxygen concentrations moderately sensitive or resistant . The flexible resistance of this aerotolerant anaerobe against metronidazole may interfere with results of sensitivity determinations, estimation of blood levels and it may influence the effectiveness of the drug in anaerobic infections due to aerotolerant anaerobes.

Aust Fam Physician, 1981 Mar, 10(3), 177 - 80
Tetanus; Feery BJ; Tetanus results from the action of a powerful exotoxin produced by Clostridium tetani . It can be prevented only by active and passive immunization procedures . The vaccine is one of the safest and most effective vaccines ever developed, and is capable of inducing long term protection in individual vaccinees . Adequate immunization before, or during pregnancy can eliminate the risk of neonatal tetanus in infants born in developing countries . In non-immune patients, tetanus prophylaxis requires the use of tetanus immunoglobulin, or tetanus antiserum in addition to adequate wound care.

Pediatrics, 1981 Mar, 67(3), 381 - 6
Antibiotic-associated pseudomembranous colitis in children; Viscidi RP et al.; Ten cases of antibiotic-associated pseudomembranous colitis in children are reviewed . The ages ranged from 4 years to 17 years; the most frequently implicated antimicrobial agents were penicillins in six children and clindamycin in two . Stool assays showed specimens from all ten patients yielded a cytopathic toxin which was neutralized by Clostridium sordellii antitoxin with titers ranging from 1:40 to 1:40,000 . Bacterial cultures of nine specimens uniformly yielded Clostridium difficile with a median concentration of 10(5.4) organisms per gram of wet weight . All nine isolates of C difficile showed a vitro production of a cytopathic toxin which was similar to or identical with that which was detected in the original stool specimen . All ten patients recovered . Six were treated with oral vancomycin and showed a good therapeutic response; one patient, however, suffered two relapses when treatment was discontinued, requiring a total of three courses of oral vancomycin . Two patients received cholestyramine and responded well . These observations provide supportive evidence that C difficile is responsible for antibiotic-associated pseudomembranous colitis in children and document efficacy of the newer therapeutic modalities in this patient population as well.

Dig Dis Sci, 1981 Mar, 26(3), 218 - 24
Bacteria and gallstones . Etiological significance; Tabata M et al.; Two hundred consecutive gallstone cases have been subjected to bacteriological study employing improved anaerobic culture techniques . In addition to Clostridia species, species of anaerobes such as Bacteroides fragilis, Peptococcus, Veillonella, and Eubacterium are found to be present, some of which possessed beta-glucuronidase activity . This finding has a certain bearing on the etiology of bile pigment calcium stones . In addition to Escherichia coli, Bacteroides and Clostridium often found in the biliary tract may contribute to the formation of bile pigment calcium stones by producing beta-glucuronidase and thus deconjugating bilirubin diglucuronide to form free unconjugated bilirubin which in turn combines with calcium, leading to stone formation . In contrast, very low bacterial incidence was associated with cholesterol stones and so-called black stones--sometimes called pure pigment stones--and thus bacteria play little role, if any, in their formation . Therefore, bile pigment calcium stones and so-called black stones, which are often classified as pigment stones indiscriminately, should be differentiated not only because of their difference in their appearance and composition but also in their etiological background . Regardless of the kind of stones present in the common duct, the incidence of bacteria was found to be increased.

Arch Microbiol, 1981 Mar, 129(1), 49 - 52
Role of hydrogen sulfide in mercury resistance determined by plasmid of Clostridium cochlearium T-2; Pan-Hou HS et al.; Mercury resistance of Clostridium cochlearium T-2P was found to be controlled by a different mechanism from those reported so far since no mercury-reducing activity was detected in this strain . The H2S generating ability as well as the demethylating activity in this bacterium was eliminated by the treatment of the cured acridine dye and recovered by the conjugation of the cured strain with the parent strain . In addition, the strain which lost their abilities to generate H2S and to decompose methylmercury, showed higher sensitivity to mercurials than the parent strain . From these results, the genes conferring both the activities seemed to reside on the plasmid and the mechanism of mercury resistance was probably based on a detoxification mechanism involving methylmercury decomposition and inactivation of the inorganic mercury with H2S.

Appl Environ Microbiol, 1981 Mar, 41(3), 737 - 45
Isolation and characterization of thirteen intestinal microorganisms capable of 7 alpha-dehydroxylating bile acids; Hirano S et al.; Thirteen anaerobic bacteria capable of performing the 7 alpha-dehydroxylation of both cholic acid and chenodeoxycholic acid were isolated from human feces and also from sewage . Ten organisms from heat-treated samples were species of Clostridium identical or closely related to the Clostridium bifermentans-C . sordellii group and consisted of four strains elaborating 7 alpha-dehydroxylase alone and six strains capable of catalyzing both 7 alpha-dehydrogenation and 7 alpha-dehydroxylation . The remaining three organisms, recovered from fresh human feces, were gram-positive, nonflagellated, nonsporeforming, anaerobic rods and comprised two distinct species . Strain HD-17, still unidentified, had both activities, but was unique in that it exclusively 7 alpha-dehydroxylated cholic acid while biotransforming chenodeoxycholic acid, preferably though 7 alpha-dehydrogenation . Two unclassified strains, b-8 and c-25, metabolized both acids though 7 alpha-dehydroxylation and 7 alpha-dehydrogenation . Except for strains b-8 and c-25, all of th 7 alpha-dehydroxylating bacteria split the conjugated bile acid series, and hydrolases were detected in cell-free filtrates of early stationary-phase broth cultures.

Ann Surg, 1981 Mar, 193(3), 361 - 4
Clostridium septicum infection and malignancy; Katlic MR et al.; Evidence mounts favoring the relationship, albeit unexplained, between Clostridium septicum infection and malignancy, particularly hematologic or intestinal malignancy . Seven patients with C . septicum gangrene or sepsis have been treated at the Massachusetts General Hospital in the years 1977-79 . All of these patients have had associated malignant disease: four patients had colon adenocarcinomas, two patients had acute myeloblastic leukemias, and one patient had breast carcinoma . In six of the seven patients, the malignancy was in an advanced state; the breast carcinoma showed no evidence of recurrence after mastectomy, 17 years earlier . A bowel portal of entry is postulated in five patients . Despite prompt use of appropriate antibiotics, the only survivors were two of the four patients who underwent early extensive debridement . These results suggest that, in the patient with C . septicum infection, malignancy should be sought; that, in the septic patient with known malignancy, C . septicum should be considered; and that, in the absence of external source in the patient with clostridial myonecrosis or sepsis, the cecum or distal ileum should be considered a likely site of infection . Increased awareness of this association between C . septicum and malignancy, and aggressive surgical treatment, may result in improvement in the present 50-70% mortality rate.

Ann Microbiol (Paris), 1981 Mar-Apr, 132(2), 149 - 57
{Detection of "Clostridium difficile" on minimal and selective medium and by immunofluorescence antibody staining (author's transl)}; Hubert J et al.; A minimal and selective medium containing 5 aminoacids, 3 vitamins, a low sugar concentration and 2 antibiotics (cefoxitin at 16 microgram/ml and streptomycin at 500 microgram/ml) is described for isolation of Clostridium difficile from the gut . Comparable results were obtained with this medium and using spore isolation by the sodium thioglycolate-lysozyme technique . An antiserum specific to C . difficile was prepared and used for detection by immunofluorescent antibody staining . This is a very sensitive technique which leads to a quick and early identification of this bacterial species.

Infect Immun, 1981 Mar, 31(3), 890 - 5
Vascular permeability activity of botulinum C2 toxin elicited by cooperation of two dissimilar protein components; Ohishi I et al.; Botulinum C2 toxin has vascular permeability as well as lethal activities . Both activities are elicited by cooperation of two dissimilar protein components, designated components I and II, which individually have very low activities . The vascular permeability activity of C2 toxin, demonstrated as blueing response after intravenous injection of Evans blue, was markedly enhanced by treatment with trypsin and was abolished by neutralization with either anti-component I or II serum . Inflammatory reactions, such as edema, congestion, and hemorrhage, were found at the site of intradermal injection of trypsinized C2 toxin . No vascular permeability activity was demonstrated by the intradermal injection of the toxin of Clostridium botulinum types A through F . These results indicate that C2 toxin has a novel biological activity, which is not possessed by the neurotoxin elaborated by C . botulinum types A through F . This suggests that C2 toxin causes lethality in a different way from that of botulinum neurotoxin, which is known to inhibit the presynaptic release of acetylcholine at the neuromuscular junction.

Appl Environ Microbiol, 1981 Mar, 41(3), 675 - 8
Nature of intracellular type A botulinum neurotoxin; Krysinski EP et al.; The neurotoxin in cells of young Clostridium botulinum type A culture was extracted with lysozyme . Highly purified neurotoxin preparation, obtained by processing the extract in two chromatographic steps had only unnicked (single-chain) molecules of molecular weight comparable to that of the dichains isolated from type A crystals . Trypsinization converted the unnicked molecules into dichains whose component subunits were of sizes indistinguishable from those of the neurotoxin from crystals . The enzymatic treatment increased toxicity of crude extract 30-fold but did not activate the purified intracellular neurotoxin preparation . The results indicated that intracellular type A botulinum neurotoxin is unnicked, is not fully activated, and is activated in the time between its extraction and purification . Since trypsinization nicked all of the single chains without increasing toxicity, nicking was not causally related to toxicity activation.

J Lipid Res, 1981 Mar, 22(3), 458 - 66
Formation of urso- and ursodeoxy-cholic acids from primary bile acids by Clostridium absonum; Macdonald IA et al.; Eight strains of Clostridium absonum were shown to form ursocholic acid (UC) from cholic acid (C) and ursodeoxycholic acid (UDC) from chenodeoxycholic acid (CDC) but did not transform deoxycholic acid (DC) in whole cell cultures . The structures of UC and UDC were verified by mass spectroscopy, and by thin-layer chromatography using Komarowsky's spray reagent . The organism transformed C and CDC at concentrations below 1.5 . 10(-3) M and 5.0 . 10(-4) M, respectively; higher concentrations were inhibitory . Optimal yields of the final products were realized at about 15-22 hr and 9-15 hr of incubation, respectively, and were in the range of 60-70% . Additionally, the 7 keto-derivatives, 7 keto-deoxycholic acid (7K-DC) or 7 keto-lithocholic acid (7K-LC) were also formed from C and CDC . With longer periods of incubation, increasing yields of 7K-DC and 7K-LC and decreasing yields of UC and UDC were observed . These time course studies suggest that 7K-DC and 7K-LC are intermediates in the formation of UC and UDC from the primary bile acids . We propose the occurrence of C right harpoon over left harpoon 7K-DC right harpoon over left harpoon UC and CDC right harpoon over left harpoon 7K-LC right harpoon over left harpoon UDC with increasing dominance of back reaction of the second step on aging of the culture . When the initial pH value of the medium was manipulated within the range of 5.8-9.0, increasing yields of UDC from CDC were obtained at higher pH values (maximum yield at pH 9.0 was 83%), with total inhibition of growth and transformation at pH 5.8 . In contrast, UC was produced from C at all pH values studied, with marginal differences in yields (maximum yield at pH 8.0 was 50%) . In all cases, formation of UC from C was much slower than that of UDC from CDC . In contrast, C . paraperfringens transformed none of the above bile acids . We propose that C . absonum, or a biochemically similar species, may be present in the human gut and give rise to UDC (and UC) in vivo.-Macdonald, I . A., D . M . Hutchison, and T . P . Forrest . Formation of urso- and ursodeoxycholic acids from primary acids by Clostridium absonum.

Arch Dermatol, 1981 Mar, 117(3), 154 - 5
Pseudomembranous colitis after topical application of clindamycin; Milstone EB et al.; Abdominal cramping and diarrhea developed in a 24-year-old woman with facial acne vulgaris five days after she started topical therapy with 1% clindamycin hydrochloride . A stool specimen contained a significant titer of a toxin produced by Clostridium difficile . Findings from sigmoidoscopy and a colonic biopsy specimen were consistent with pseudomembranous colitis . The patient became asymptomatic after ten days of supportive care and oral vancomycin hydrochloride therapy . This case is presented as an example of pseudomembranous colitis associated with topical application of clindamycin.

J Clin Microbiol, 1981 Mar, 13(3), 498 - 502
Optimal fluorescein-to-protein ratios of bacterial direct fluorescent-antibody reagents; Hebert GA et al.; A number of bacterial systems were studied with specific direct fluorescent-antibody reagents prepared from rabbit antiserum fractions and having a wide range of fluorescein-to-protein ratios . These systems included Bacteroides, Bordetella, Clostridium, Escherichia, Legionella, Listeria, Salmonella, Shigella, and Streptococcus . For all systems studied, a fluorescein-to-protein ratio of 30 was optimal for conjugates prepared from ammonium sulfate fractions (greater than 75% gamma globulin) and pure immunoglobulin G desorbed from the Sepharose-bound protein A of Staphylococcus aureus . A pepsin digestion procedure is described that yielded the F(ab')2 piece of pure immunoglobulin G; this was labeled and studied at two fluorescein-to-protein ratios.

Biochim Biophys Acta, 1981 Feb 27, 667(2), 433 - 51
Low-temperature magnetic circular dichroism spectra and magnetisation curves of 4Fe clusters in iron-sulphur proteins from Chromatium and Clostridium pasteurianum; Johnson MK et al.; The magnetic circular dichroism (MCD) spectra of the 4Fe clusters in the iron-sulphur proteins high-potential iron protein from Chromatium and the 8Fe ferredoxin from Clostridium pasteurianum have been measured over the wavelength range 300-800 nm at temperatures between approx . 1.5 and 50 K and at magnetic fields up to 5 tesla . In both cases the proteins have been studied in the oxidized and reduced states . The reduced state of high-potential iron protein gives a temperature-independent MCD spectrum up to 20 K, confirming the diamagetism of this state at low temperature . The MCD spectrum of samples of oxidized ferredoxin invariably show the presence of a low concentration of a paramagnetic species, in agreement with the observation that the EPR spectrum always shows a signal at g = 2.01 . The paramagnetic MCD spectrum runs across the whole of the wavelength range studied and therefore most probably originates from an iron-sulphur centre . The diamagnetic component of the MCD spectrum of oxidized ferredoxin is very similar to that of reduced high-potential iron protein . The low-temperature MCD spectra of oxidized high-potential iron protein and reduced ferredoxin reveal intense, temperature-dependent bands . The spectra are highly structured with that of high-potential iron protein showing a large number of electronic transitions across the visible region . The MCD spectra of the two different oxidation levels are quite distinctive and should provide a means of establishing the identity of these state of 4Fe clusters in more complex proteins . MCD magnetisation curves have been constructed from detailed studies of the field and temperature dependence of the MCD spectra of the two paramagnetic oxidation states . These plots can be satisfactorily fitted to the theoretically computed curves for an S = 1/2 ground state with the g factors experimentally determined by EPR spectroscopy . The low-temperature MCD spectra of the reduced 2Fe-2S ferredoxin from Spirulina maxima are also presented and MCD magnetisation curves plotted and fitted to the experimentally determined g factors.

J Biol Chem, 1981 Feb 25, 256(4), 1968 - 74
Isolation and structural characterization of human lymphocyte and neutrophil gangliosides; Macher BA et al.; Gangliosides were isolated from purified preparations of human peripheral blood lymphocytes and neutrophils . Structural analyses and comparisons were performed by direct probe mass spectrometry and by degradation studies with the following enzymes: Escherichia freundii endo-beta-galactosidase; Clostridium perfringens and Arthrobacter ureafaciens neuraminidase; and jack bean beta-N-acetylhexosaminidase and beta-galactosidase . This combination of techniques allowed us to obtain carbohydrate composition and sequence information without the aid of methylation or carbohydrate compositional analyses using only 1-2 mg of purified gangliosides . On the basis of these studies we propose that human lymphocytes and neutrophils have gangliosides with the following structures . NeuAc alpha 2 leads to 3Gal beta 1 leads to 4Glc beta 1 leads to 1Cer Structure A NeuAc alpha 2 leads to ? GlcNAc beta 1 leads to 3Gal beta 1 leads to 4Glc beta 1 leads to 1Cer Structure B NeuAc alpha 2 leads to ? Gal beta 1 leads to 3,4GlcNAc beta 1 leads to 3Gal beta 1 leads to 4Glc beta 1 leads to 1Cer Structure C All three compounds were isolated from both cell types with structure A being the major lymphocyte ganglioside and structure C the major neutrophil ganglioside . Structure B is a novel ganglioside and may represent a leukocyte-specific glycosphingolipid . Neuraminidase degradation studies demonstrated that only one ganglioside species of each cell type contains an internally linked sialic acid residue, and on the basis of thin layer chromatographic analysis this component is the same as the major brain ganglioside, GM1 (II3-N-acetylneuraminosyl-gangliotetraosylceramide) . In addition, large gangliosides with the general structure NeuAc alpha 2 leads to ?(Gal beta 1 leads to 3,4GlcNAc beta 1 leads to 3)n Gal beta 1 leads to 4Glc beta 1 leads to 1Cer were isolated . These results are discussed as they relate to blood group antigens and specific cell surface markers in human leukocytes.

Biochim Biophys Acta, 1981 Feb 20, 641(1), 138 - 47
Some properties of a new electrogenic transport system: the ammonium (methylammonium) carrier from Clostridium pasteurianum; Kleiner D et al.; Clostridium pasteurianum is able to build up about 100-fold gradients of methylammonium across the cell membrane . Methylammonium enters the cell by means of a carrier as shown by the energy requirement, saturation kinetics and a pH profile with a narrow maximum between pH 6.2 and 6.8 . The methyl ammonium transport (apparent Km = 150 microM, V = 100 mumol/min per g dry weight) is competitively inhibited by ammonium (apparent Ki = 9 microM) . The low Ki value and the observation that methylammonium cannot serve as a carbon or nitrogen source for Cl . pasteurianum strongly indicate that ammonium rather than methylammonium is the natural substrate . Uncouplers and inhibitors of energy metabolism or of the membrane-bound ATPase inhibit transport . Cl . pasteurianum maintains a membrane potential (interior negative) in the range 80-130 mV . This membrane potential was identified as the energy source: the same agents that block transport also decrease the membrane potential, and artificial generation of a membrane potential (by addition of valinomycin to K+-loaded cells) induces concentrative uptake of methylammonium . Thus NH4+ (or CH3NH3+) must be the transported species . Digestion of the cell wall by lysozyme does not abolish the transport activity.

Lancet, 1981 Feb 14, 1(8216), 371 - 2
Is pseudomembranous colitis infectious?
Greenfield C, Burroughs A, Szawathowski M, Bass N, Noone P, Pounder R.
A cluster of eight patients in two adjacent hospital wards acquired acute diarrhoea within a period of 11 days . All their stool samples contained Clostridium difficile toxin and C . difficile was isolated in every case . Three patients had rectal biopsy findings compatible with pseudomembranous colitis (PMC) . All the patients responded to treatment with oral vancomycin . Until the possibility of PMC being acquired by cross-infection is clarified such patients should be nursed in isolation with strict enteric precautions.

J Clin Pathol, 1981 Feb, 34(2), 217 - 20
Non-toxigenic clostridia in babies; Mitchell RG et al.; Non-toxigenic clostridia, Clostridium butyricum but occasionally C . paraputrificum, were isolated from mainly extra-alimentary sites of 28 babies in maternity hospital over a period of 40 weeks . During that time, C . butyricum was isolated from the blood of two babies who died of necrotising enterocolitis and of two babies without referable symptoms . Otherwise these organisms appeared to be of no pathogenic significance.

Can J Microbiol, 1981 Feb, 27(2), 216 - 25
Growth inhibition activity and bacteriophage and bacteriocinlike particles associated with different species of Clostridium; Nieves BM et al.; Thirty-four strains corresponding to 16 species of Clostridium were examined for growth inhibition activity and production of phage and bacteriocinlike particles . Twenty-five strains were found to inhibit growth of one or more of the other strains . The widest range of activity corresponded to C . bifermentans 18137 which inhibited growth of 23 strains . Cell-free filtrates of positive cultures were inactive with the exception of the one from C . bifermentans 18137 . A wide variety of particles with diverse morphology was observed in lysates of cultures treated with mitomycin C (1 microgram/mL).

Appl Environ Microbiol, 1981 Feb, 41(2), 375 - 80
Nisin: a possible alternative or adjunct to nitrite in the preservation of meats; Rayman MK et al.; Nisin at 75 ppm (75 microgram/g) was superior to 150 ppm of nitrite in inhibiting outgrowth of Clostridium sporogenes PA3679 spores in meat slurries, which had been heated to simulate the process used for cooked ham . The inhibitory activity of nisin decreased as the spore load or pH of the slurries increased . Unlike nitrite, inhibition by nisin was unaffected by high levels of iron either as a constituent of meats or when added as an iron salt . In slurries treated with 75 ppm of nisin, refrigerated storage for 56 days resulted in depletion of nisin to a level low enough to allow outgrowth within 3 to 10 days if the slurries were subsequently abused at 35 degrees C . In contrast, a combination of 40 ppm of nitrite and either 75 or 100 ppm of nisin almost completely inhibited outgrowth in these slurries . The nisin-nitrite combination appeared to have a synergistic effect, and the low concentration of nitrite was sufficient to preserve the color in meats similar to that of products cured with 150 ppm of nitrite.

Am J Med, 1981 Feb, 70(2), 432 - 8
Bacteria newly recognized as nosocomial pathogens; Fraser DW; Bacteria recently recognized as nosocomial pathogens generally fall into three categories: those that grow slowly, those that are fastidious in their nutritional or atmospheric requirements and those that resemble commensals . Each characteristic has contributed to the delay in perceiving their importance . Mycobacterium chelonei and Myco . fortuitum--which grow slowly, although characterized as "rapid-growing" mycobacteria--cause sternal osteomyelitis, pericarditis and endocarditis after cardiac surgery as well as other wound infections after many types of surgery . Myco . chelonei-like organisms have been found to cause "sterile" peritonitis in patients receiving long-term peritoneal dialysis . Legionella pneumophila and L . micdadei are fastidious bacteria that were more difficult to detect because they stain poorly with the Gram method . They cause pneumonia and lung abscess, especially in immunocompromised people . Clostridium difficile is an anaerobe that causes toxin-mediated pseudomembranous colitis in persons given antibiotics that inhibit competing gut bacteria . Chylamydia trachomatis, an intracellular organism that has not been grown in vitro, causes pneumonia and conjunctivitis in young infants who acquire the organism from their mothers at birth . Group JK bacteria cause septicemia in patients whose immune responses have been suppressed and must be distinguished from "diphtheroid" contaminants in blood cultures . Clinicians, microbiologists and epidemiologists must be alert to the characteristics of these organisms that make them easily overlooked and should also anticipate the existence of other bacteria not yet identified.

Appl Environ Microbiol, 1981 Feb, 41(2), 539 - 41
Carboxymethyl cellulase and cellobiase production by Clostridium acetobutylicum in an industrial fermentation medium; Allcock ER et al.; The production of a carboxymethyl cellulase and a cellobiase by Clostridium acetobutylicum was demonstrated . In liquid medium the carboxymethyl cellulase was induced by molasses, and it was not repressed by glucose . Optimum carboxymethyl cellulase activity occurred at pH 4.6 and 37 degrees C.

Antimicrob Agents Chemother, 1981 Feb, 19(2), 274 - 8
Macrolide-lincosamide-streptogramin resistance patterns in Clostridium perfringens from animals; Dutta GN et al.; Different patterns of resistance against commonly used macrolide, lincosamide, and streptogramin antibiotics were found in Clostridium perfringens of animal origin . The patterns were designated as (i) macrolide-lincosamide-streptogramin group B generalized resistance, (ii) macrolide-lincosamide generalized resistance, (iii) macrolide-lincosamide inducible resistance, and (iv) macrolide-lincosamide-streptogramin low-level generalized resistance . The strains of the fourth pattern were able to inactivate pristinamycin and virginiamycin . The macrolide-susceptible strains showed a bimodal distribution of lincomycin and clindamycin susceptibility levels . The susceptible strains were inhibited by 0.25 micrograms of lincomycin per ml and 0.03 micrograms of clindamycin per ml . The low-level resistant strains were inhibited at concentrations of 2 to 4 micrograms of lincomycin per ml and 0.5 to 2 micrograms of clindamycin per ml.

Infect Immun, 1981 Feb, 31(2), 608 - 14
Common polysaccharide antigens from the cell envelope of Clostridium perfringens type A; Dayalu KI et al.; Soluble antigens were obtained by extracting five serotype strains of Clostridium perfringens type A with water at 100 degrees C . The type-specific polysaccharides were precipitated with ethanol, and the common antigens were recovered from the ethanol supernatants by concentration, dialysis, and lyophilization . Refluxing the water-extracted cell residues with 1% acetic acid followed by concentration, dialysis, and lyophilization gave additional common antigen fractions . A comprehensive, side-by-side comparison of the antigen fractions, the ethanol precipitate, the ethanol supernatant, and the acetic acid supernatant, revealed that common antigens were recovered in all three fractions, and that three distinct entities were responsible for the formation of the observed common immunoprecipitin lines; whereas many fractions possessed all three immunoprecipitin lines, others contained only one or two . The serological homology observed between the various antigen fractions was apparently a consequence of N-acetylglucosamine- and N-acetylmannosamine-containing polymers . The common antigens were presumably associated with the cell envelope and may be the type of markers sought previously by others for the serological identification of C . perfringens.

Infect Immun, 1981 Feb, 31(2), 536 - 46
Purification and characterization of Clostridium perfringens delta-toxin; Alouf JE et al.; Delta-toxin, an extracellular hemolysin released by Clostridium perfringens type C, was purified from culture supernatant fluid by sequential ammonium sulfate precipitation, thiol-Sepharose gel chromatography, isoelectric focusing, and Sephadex G-75 gel filtration . The purified preparation had a specific activity of 320,000 hemolytic units per mg of protein and was homogeneous, as determined by immunochemical and electrophoretic tests . This toxin was characterized as a single polypeptide chain composed of 391 amino acid residues, 30% of which were hydrophobic . The molecular weight was found to be 42,000, and the isoelectric point was pH 9.1 . Delta-toxin appeared to be amphiphilic by charge shift electrophoresis in a three-detergent system . It was immunogenic in rabbits and lethal to mice at a dose of 0.12 micrograms . The lytic activity of delta-toxin was restricted to erythrocytes of even-toed ungulates (sheep, goats, and pigs) . This activity was inhibited by GM2 ganglioside but not by other gangliosides, cholesterol, lecithin, or sphingomyelin.

Med Biol, 1981 Feb, 59(1), 11 - 20
The action of batulinum toxin at the neuromuscular junction; Sellin LC; Botulism results from the action of a protein neurotoxin (mol . wt . congruent to 150,000) produced by the bacterium Clostridium botulinum, of which there are eight known strains . Botulinum neurotoxin is the most potent biological toxin known, having a median lethal dose of 5--50 ng/kg body weight . The primary site of action of botulinum toxin is the cholinergic nerve terminal, where it blocks the release of the neurotransmitter acetylcholine . Death usually results from respiratory failure . Nonlethal doses of botulinum toxin can induce sprouting of the nerve terminal and have significant postsynaptic effects, including muscle atrophy and alterations in the membrane electrical properties of the muscle fiber . There is no universally available treatment for botulinum intoxication . However, immunotherapeutic and chemotherapeutic procedures are not being developed and will be discussed.

Med Clin (Barc), 1981 Jan 25, 76(2), 70 - 2
{Spontaneous peritonitis due to Clostridium perfringens in three patients with cirrhosis (author's transl)}; Moya Mir MS et al.; Three cases of spontaneous peritonitis due to Clostridium perfringens in cirrhotic patients with a fatal outcome are reported . The diagnosis was made clinically in two patients and by post-mortem examination in the third . One patient had elevated values of serum alpha-fetoprotein . These cases are compared with three other reported cases in the literature . Blood cultures were negative in the three patients, a fact that lends support to the theory of transmural migration of bacteria . In the authors' experience C . perfringens is the third most frequent agent responsible for spontaneous peritonitis in cirrhosis, preceded by E . coli and Streptococcus and followed by Klebsiella, a surprising fact given the scarce number of reported cases . Routine abdominal paracentesis is recommended in any cirrhotic patient with ascites, followed by appropriate antibiotic treatment whenever positive cultures are obtained . The efficacy of treatment is probably doubtful . The literature on antibiotic treatment of spontaneous peritonitis in cirrhosis is reviewed.

C R Seances Acad Sci III, 1981 Jan 19, 292(3), 285 - 7
{Penicillin resistance due to the production of a beta-lactamase in Clostridium butyricum}; Magot M; Eight strains of clostridia of the butyricum group, resistant to penicillins were isolated from perinatal infections . MIC's of these strains vary from 1,000 to 3,000 micrograms/ml for benzyl-penicillin, and from 5 to 10 micrograms/ml for cefoxitin . The substrate profile of the enzymes is that of a penicillinase.

J Biol Chem, 1981 Jan 10, 256(1), 165 - 9
Isolation and characterization of a novel monosialosylpentahexosyl ceramide from Tay-Sachs brain; Itoh T et al.; A novel monosialoganglioside was isolated from Tay-Sachs brains . It represented about 0.1% of the total ganglioside mixture . Compositional analysis by gas-liquid chromatography indicated that it contained glucose, galactose, N-acetylgalactosamine, N-acetylneuraminic acid, and long chain base in the molar ratio of 1:2:2:1:1 . The ganglioside was found to be resistant to neuraminidase (Clostridium perfringens), beta-hexosaminidase (jack bean), and beta-galactosidase . However, it could be degraded by a human liver beta-hexosaminidase preparation in the presence of an activator to produce a glycolipid chromatographically identical with authentic IV3NeuAc-GgOse4-ceramide . This glycolipid product was resistant to beta-galactosidase (jack bean), but could be readily degraded to GgOse4-ceramide by neuraminidase . Mild formic acid hydrolysis degraded the intact ganglioside to an asialo derivative chromatographically identical with the pentahexosyl ceramide (GalNAc-Gal-GalNAc-Gal-Glc-ceramide) derived from GD1a-GalNAc . The asialo derivative could then be degraded to GgOse4-ceramide and GgOse3-ceramide by sequential treatment with jack bean beta-hexosaminidase and beta-galactosidase . These data suggest that the novel ganglioside is a monosialosylpentahexosyl ceramide with the sialosyl group attached to the penultimate galactose moiety of the pentahexosyl ceramide backbone, and it has the following structure: GalNAc(beta 1-4)Gal(beta 1-3)GalNAc(beta 1-4)Gal(beta 1-4)Glc-ceramide (formula see text).

Med Clin (Barc), 1981 Jan 10, 76(1), 23 - 5
{Clostridium septicum bacteremia in a diabetic patient (author's transl)}; Asencio Marchante R et al.; This report concerns the clinical and bacteriological study of a female diabetic who presented with a febrile illness of sudden onset and acute toxemia . Physical examination at admission was unrewarding . A few hours after admission lesions characteristic of gas gangrene appeared in the right upper limb without prior trauma, and a clostridial infection was suspected . Bacteriological studies of the exudate and of the patient's blood led to the isolation of Clostridium septicum, thus confirming the clinical suspicion . The disease had a precipitous course and the patient died 12 hours after admission in spite of the administration of high doses of intravenous penicillin . While waiting for bacteriological data, early antibiotic treatment appears to be indicated in any diabetic patient with an acute toxic syndrome and absence of localized infection.

Gut, 1981 Jan, 22(1), 34 - 7
Spectrum of antibiotic-associated diarrhoea; Lishman AH et al.; In an attempt to find the extent to which Clostridium difficile could be implicated as the cause of antibiotic-associated diarrhoea, the stools of 53 patients who had diarrhoea after a course of antibiotics were investigated for the presence of C . difficile toxin . Ten of the patients (19%) were found to be positive, but the stools of four out of 53 patients without diarrhoea after a course of antibiotics were also found to contain C . difficile toxin (7.5%) . The titre of toxin in patients both with and without diarrhoea fell within the same range (up to 10(-5)) . Neither the organism nor its toxin was found in the stool of 26 patients with ulcerative colitis, eight with Crohn's disease, 49 with non-specific diarrhoea, and 27 normal controls . We conclude that, while C . difficile is responsible for a proportion of cases of antibiotic-associated diarrhoea, the concentration of toxin is not the sole factor affecting the severity of this disorder.

Br J Surg . 1981 Jan;68(1):38.
Gas gangrene in intestinal angina; McLenachan J et al.; A patient with Clostridium welchii infection complicating intestinal angina is reported . Surgical resection of bowel of dubious viability is important to avoid this unusual complication.

Clin Exp Neurol, 1981, 17, 147 - 51
Subacute cholinergic dysautonomia in childhood; Hopkins IJ et al.; Clinical features of a 10-year-old boy who presented with manifestations of subacute autonomic neuropathy are described . The pupillary abnormalities, reduced tear, sweat and saliva production, and visceral dysfunction suggest a lesion restricted to cholinergic postganglionic endings of the autonomic nervous system . There was no evidence of dysfunction at the neuromuscular junction or elsewhere in the nervous system . The cause of the cholinergic dysautonomia has not been found . Clostridium botulinum infection could not be verified, and although a variety of Clostridium species were isolated from the faeces they could not be shown to produce a neurotoxin.

Med Microbiol Immunol (Berl), 1981, 170(1), 27 - 35
Occurrence of toxin-producing Clostridium difficile in antibiotic-associated diarrhea in Sweden; Aronsson B et al.; From 1324 patients with antibiotic-associated diarrhea (AAD) 1643 stool samples were analyzed by a cell test for Clostridium difficile toxin in stool filtrates and cultivation for occurrence of C . difficile strains . In patients with no detectable toxin in their stool strains of C . difficile were isolated in 2.2% whereas when toxin was detectable, the isolation rate varied from 17% to 36% . Furthermore, there was a correlation between toxin titre in stool filtrate and production of cytotoxin in vitro by the corresponding C . difficile strains . Five clostridial strains, not belonging to the species C . difficile, were found to produce typical cytotoxin in vitro . However, five strains identified as C . difficile by biochemical reactions and gas liquid chromatography, did not produce an extracellular cytotoxin . The antibiotic susceptibility patterns of the Clostridium strains were investigated . No correlation was recognized between antibiotic resistance of isolated Clostridium strains and the AAD-inducing antibiotic penicillins and linco/clindamycin . Neither did cases of relapse of diarrheal disease after vancomycin treatment harbour C . difficile strains with increased resistance to vancomycin . It is concluded that the pathogenesis of antibiotic-associated enterocolitis is more complex than a mere intestinal overgrowth of resistant strains of C . difficile.

Avian Dis, 1981 Jan-Mar, 25(1), 184 - 94
Naturally occurring and experimentally induced castor bean (Ricinus communis) poisoning in ducks; Jensen WI et al.; Castor bean (Ricinus communis) poisoning accounted for the death of several thousand ducks in the Texas panhandle in the fall and winter months of 1969-1971 . Signs of intoxication resembled those of botulism, except for mucoid, blood-tinged excreta . The most common lesions were severe fatty change in the liver, widely distributed internal petechial hemorrhages or ecchymoses, and catarrhal enteritis . Nearly intact castor beans were found in the stomach of one duck during field necropsy . Fragments of seed coat resembling castor bean were found in the stomachs of 10 of 14 ducks examined in the laboratory . Clinical signs and postmortem lesions observed in wild ducks were induced experimentally in mallards (Anas platyrhynchos) by force-feeding intact castor beans . Toxicity titrations were erratic, but the LD50 appeared to be between three and four seeds . The mouse toxicity test, used to detect Clostridium botulinum toxin in the blood serum of intoxicated ducks, was negative in every case . Hemagglutination and precipitin tests generally failed to detect castor bean in extracts of excreta or intestinal contents of experimentally intoxicated ducks.

J Infect Dis, 1981 Jan, 143(1), 51 - 4
Studies on the epidemiology of colitis due to Clostridium difficile in hamsters; Toshniwal R et al.; Hamsters treated with vancomycin developed enterocolitis significantly more often in a conventional animal room than in a room designed to prevent cross-infection with Clostridium difficile . In the conventional room C . difficile was isolated from cages, food racks, floors, buckets, the hands of caretakers, and the stools of animals with enterocolitis but not from untreated hamsters, air, or food from freshly opened bags . C . difficile was not isolated from environmental sources in the clean room . It was not possible to determine which of the sources of the organism was most important in its spread . Cross-infection with C . difficile may be important in the pathogenesis of antibiotic-associated enterocolitis in hamster colonies.

Hoppe Seylers Z Physiol Chem, 1981 Jan, 362(1), 33 - 8
Stereospecific reductions of 2-en-1-ols catalyzed by Clostridium kluyveri; Angermaier L et al.; With ethanol as electron donor, resting cells of Clostridium kluyveri reduced (E)-2-methyl-2-buten-1-ol, (E)-3-methyl-2-penten-1-ol and (E)-2-methyl-3-phenyl-2-propen-1-ol to (R)-2-methyl-1-butanol, (R)-3-methyl-1-1-pentanol and (R)-2-methyl-3-phenyl-1-propanol, respectively . Within the experimental errors of ORD measurements the products were optically pure . That means that only one of the two possible trans additions to the carbon-carbon double bond of these derivatives of allyl alcohol took place . The reduction of (E)-2-methyl-3-phenyl-2-propen-1-ol was carried out in 2H2O buffer . Assuming tha all 3 substrates bind in an identical manner to the enzyme, the product should be (2R,3S)-2-methyl-3-phenyl-{2,3-2H2}-1-propanol . The optimal pH and ethanol concentration for the reduction have been determined . Under an atmosphere of hydrogen the reduction of the unsaturated alcohols proceeded incompletely and slower than in the presence of ethanol under an atmosphere of nitrogen.

Acta Paediatr Scand, 1981 Jan, 70(1), 129 - 30
Case report . Clindamycin associated pseudomembranous colitis; Beesley J et al.; Pseudomembranous colitis is rare in children . We describe a case associated with clindamycin in which Clostridium difficile and its enterotoxin were isolated from the stool . Treatment with oral vancomycin brought about a prompt and complete recovery.

Arch Geschwulstforsch, 1981, 51(1), 51 - 7
Cocultivation of Clostridium oncolyticum with normal and tumour cell lines; Schlechte H et al.; During incubation of monolayer cell cultures with clostridia spores in the presence of 1.5 per cent of oxygen vegetative bacterial rods become visible and cell necrosis occurs preferentially in tumour cell lines . After such cocultivation tumour cells show a higher amount of clostridia not elutable by washing than finite cells . Electron microscopy of associated clostridia shows vegetative rods in contact to and inside of tumour cells.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1981, 250(1-2), 142 - 6
Simple method for isolation and presumptive identification of Clostridium difficile; Al-Jumaili IJ et al.; Clostridium difficile can be isolated from stools and presumptively identified by inoculating the stools onto CCCBA and the culture looked at under ultra violet light where the golden-yellow fluorescent colonies show up against a dark background . The colonies are then identified by testing them using the API ZYM system and the results further confirmed by testing the culture for toxin production using the tissue culture technique employing Cl . sordellii antitoxin.

Zentralbl Bakteriol Mikrobiol Hyg {A}, 1981, 249(4), 512 - 9
Measurement of specific multiplication of an oncolytic and a nononcolytic strain of Clostridium butyricum inside the tumour UVT 15264 of the mouse; Schlechte H et al.; Clostridium butyricum was recultivated from tumour material after treatment of tumour-bearing mice with spores and counted by soft agar colony technique . The oncolytic strain Cl . butyricum H8 grows inside the tumour UVT 15264 up to 8 X 10(8)/g during 48 h . In this time part of tumour became lysed . Multiplication inside of this tumour by the tumour specific but nononcolytic strain Cl . butyricum CNRZ 528 could not be detected by application counting technique . Combination of spore treatment with antibiotic chemotherapy in dosage able to prevent oncolysis or in dosage to reduce them corresponds with loss of clostridial multiplication or with reduced vegetative multiplication respectively.

Appl Environ Microbiol, 1981 Jan, 41(1), 179 - 83
Persistence of Clostridium botulinum type B on a cattle farm after an outbreak of botulism; Notermans S et al.; On farms involved in botulism outbreaks, cycles of Clostridium botulinum have occurred . The cycles were initiated by feeding brewers' grains contaminated with proteolytic C . botulinum type B to the cows . Spreading of manure containing feces of these cows increased the contamination of the pastures . In grass silages prepared with wilted grass from these pastures the number of C . botulinum type B organisms increased, and toxin type B was produced . Feeding cows with the contaminated silage fodder completed the cycle . Besides contamination of human foodstuffs (milk and meat), further contamination of the environment occurred . It was demonstrated that fowl may be important vectors in spreading C . botulinum.

J Infect Dis, 1981 Jan, 143(1), 22 - 7
Isolation of Clostridium botulinum type G and identification of type G botulinal toxin in humans: report of five sudden unexpected deaths; Sonnabend O et al.; Clostridium botulinum type G has not been identified until now from humans or animals; it has been isolated only twice, from soil samples in Argentina . Type G organisms were isolated from necropsy specimens in four adults and an 18-week-old infant . Type G botulinal toxin was demonstrated in the serum of three of these individuals . The toxic dose in mice ranged from 2 to 7 50% lethal doses/ml . These persons died suddenly and unexpectedly at home, without any pathologic evidence to account for the cause of death in four cases . Symptoms in two individuals were similar to those observed in food-borne botulism . Thus, a prompt postmortem search for toxin and organisms of C . Botulinum in blood and feces may be worthwhile in determining the etiology of unexplained deaths . More microbiologic, physiologic, and toxicologic data are needed to clarify the role of C . botulinum in the pathogenesis of sudden unexpected death in infants and adults.

Arch Microbiol, 1981 Jan, 128(3), 341 - 2
Relative levels of guanosine 5'-diphosphate 3'-diphosphate (ppGpp) in some N2 fixing bacteria during derepression and repression of nitrogenase; Kleiner D et al.; Addition of ammonium to N2 fixing cultures of Azotobacter vinelandii, Klebsiella pneumoniae and Clostridium pasteurianum rapidly reduced the intracellular levels of guanosine 5'-diphosphate 3'-diphosphate (ppGpp) by 70-90% . This change might reflect a regulatory role of ppGpp in nitrogen metabolism.

Ann Biol Clin (Paris), 1981, 39(1), 1 - 8
{Role of Clostridium and its toxin in pseudo-membranous colitis (author's transl)}; Bryskier A et al.; At present many authors consider that pseudo-membranous colitis is of bacterial origin . The main pathogenic agent is Clostridium difficile . It is not easy to isolate this organism in the stool, selective media are under study . It liberates a lipo-glycoprotein exotoxin during lysis . It is only partially purified, its structure is not fully elucidated . Its molecular weight is not yet precisely determined . It consists of several polymerised polypeptide fragments of molecular weight 50 000 . It is a thermolabile acid and alkaline sensitive cytotoxin which acts on the cell membranes and the ileo-caeco-colonic mucosa of man and animals . Clostridium difficile is transmissible by a small number of high risk carrier subjects who are potentially patients with pseudo-membranous colitis . Antibiotic therapy may lead to unbalance of the ecosystem represented by the bacterial flora of the digestive tract and favour the multiplication of a resistant strain to the administered antibiotic . The appearance of pseudo-membranous colitis requires the association of sufficient bacterial development (equal or greater than 10(7) germs per gram of stools) and the liberation of a cytotoxin . The pathogenic treatment consists of antibiotic therapy by Vancomycin or Metronidazole which seems, at present, the most active on the germs and a toxin absorbent, such as Cholestyramine, Coliptol hydrochloride or Heavy metals.

Scand J Infect Dis Suppl, 1981, 26, 9 - 13
The role of anaerobes in human infections; Finegold SM; Anaerobic bacteria have been shown to play a role in infection of all types in humans . Certain infections are notable for the prominent role played by anaerobes; included are brain abscess, chronic sinusitis and otitis media, oral and dental infections, neck space infections, bite infections, lung abscess, aspiration pneumonia, empyema, intra-abdominal infections of all types (notably peritonitis, intra-abdominal abscess, and liver abscess), abdominal surgical wound infections, female genital tract infections of all types, various superficial and deep soft tissue infections, and osteomyelitis . In recent years, two new anaerobic infections have been appreciated-infant botulism and pseudomembranous colitis due to Clostridium difficile . Considerable progress has been made recently in delineating factors predisposing to anaerobic infections, virulence factors in anaerobes, and host defense mechanisms vs . these organisms . Taxonomic schemes for anaerobes have been improved and simplified considerably, as have techniques for growing and identifying anaerobic bacteria . Rapid procedures are coming into the picture . Finally, much has been learned about therapy of these infections . Although there has been some problem of increased resistance of anaerobes to antimicrobial agents, several promising new drugs are now available to offset this.

Scand J Infect Dis Suppl, 1981, 26, 46 - 53
Nitroimidazoles: in vitro activity and efficacy in anaerobic infections; Tally FP et al.; Nitroimidazoles, including metronidazole, tinidazole and ornidazole, are low molecular weight antimicrobial compounds with excellent activity against anaerobic microorganisms . These compounds are usually bactericidal at low concentrations and their spectrum of activity encompasses almost all the anaerobic bacteria and some capnophylic organisms . The few anaerobic bacteria known to be resistant to the nitroimidazoles include occasional anaerobic cocci, some nonsporing gram positive bacilli and Propionibacterium . Nitroimidazoles are the most active antimicrobial agents known against Bacteroides fragilis, the most resistant of anaerobic bacteria . Kill-curve studies demonstrate that there is a 2 to 5 log decrease in the number of colony forming units with Bacteroides fragilis and clostridium perfringens within one hour . The killing is unaffected by inoculum, growth rate or components of the medium . However, a metronidazole resistant isolate of B . fragilis has been shown to have decreased ability to take up 14C-metronidazole as well as lessened ability to reduce metronidazole . This is associated with a decrease in the nitroreductase activity . The in vitro observations have also been demonstrated in vivo . Clinical studies have shown nitroimidazoles to be efficacious in the therapy of a variety of anaerobic infections including non-traumatic brain abscess, intraabdominal abscesses, pelvic suppuration and necrotizing soft tissue infections . However, there have been disappointing results in the therapy of anaerobic pleuropulmonary infections, with a number of superinfections caused by aerobic bacteria.

Scand J Infect Dis Suppl, 1981, 26, 42 - 5
In vitro susceptibility of anaerobic bacteria to nitroimidazoles; Olsson-Liljequist B et al.; The in vitro susceptibility of 355 Swedish clinical isolates of anaerobic bacteria to three nitroimidazole drugs was determined by the agar dilution method . The drugs were metronidazole, tinidazole and ornidazole . In addition the in vitro susceptibility of 32 strains of clostridium difficile to metronidazole was determined . Gram-negative rods and clostridia were inhibited by 4 microgram/ml or less of all three compounds except one strain of C . difficile, which was not inhibited by 16 microgram/ml of metronidazole . Against strains of the Bacteroides fragilis group, tinidazole was slightly more active than metronidazole and ornidazole . Only about 70% of anaerobic cocci (peptococci and peptostreptococci) were inhibited by 4 microgram/ml of the three drugs, whereas the remaining isolates were resistant (minimum inhibitory concentration greater than or equal to 8 microgram/ml) . Anaerobic streptococci were not included in this material . Actinomyces, arachnia and propionibacteria were resistant to the nitroimidazole drugs . At achievable serum levels, nitroimidazoles inhibited the majority of anaerobic bacteria found with greatest frequency in anaerobic infections.

J Gen Microbiol, 1981 Jan, 122(Pt 1), 41 - 6
Immunological analysis of the EDTA-soluble antigens of Clostridium difficile and related species; Poxton IR et al.; Antigens were extracted with EDTA from 32 strains representing 10 species of Clostridium . When these antigens were examined by an enzyme-linked immunosorbent assay, marked cross-reactions were observed between C . difficile, C . sordellii and C . bifermentans . The cross-reactive antigen, visualized by crossed immunoelectrophoresis, was carbohydrate.

Am J Pediatr Hematol Oncol, 1981 Spring, 3(1), 87 - 8
Successful therapy of Clostridium septicum sepsis in a child with Burkitt's lymphoma; Toledano SR et al.; The association between Clostridium septicum sepsis in patients with malignant disease has been frequently documented . A presentation with fever, neutropenia, vomiting, and an acute abdomen is characteristic of this anaerobic infection which has been uniformly fatal in children with acute leukemia . We report the unusual course and the successful treatment of an adolescent with an abdominal Burkitt's lymphoma with leukemic transformation and clostridium septicum sepsis and cellulitis.

Arch Microbiol, 1981 Jan, 128(3), 294 - 8
Effect of molecular hydrogen and carbon dioxide on chemo-organotrophic growth of Acetobacterium woodii and Clostridium aceticum; Braun K et al.; During growth of Acetobacterium woodii on fructose, glucose or lactate in a medium containing less than 0.04% bicarbonate, molecular hydrogen was evolved up to 0.1 mol per mol of substrate . Under an H2-atmosphere growth of A . woodii with organic substrates was completely inhibited whereas under an H2/CO2-atmosphere rapid growth occurred . Under these conditions H2 + CO2 and the organic substrate were utilized simultaneously indicating that A . woodii was able to grow mixotrophically . Clostridium aceticum differed from A . woodii in that H2 was only evolved in the stationary phase, that the inhibition by H2 was observed at pH 8.5 but not at pH 7.5, anf that in the presence of fructose and H2 + CO2 only fructose was utilized . The hydrogenase activity of fructose-grown cells of C . aceticum amounted to only 12% of that of H2 + CO2-grown cells . With A . woodii a corresponding decrease of the activity of this enzyme was not observed.

Arch Microbiol, 1981 Jan, 128(3), 288 - 93
Clostridium aceticum (Wieringa), a microorganism producing acetic acid from molecular hydrogen and carbon dioxide; Braun M et al.; Spores of Clostridium aceticum (Wieringa) from 1947 have been revived, and a study for a redescription of this microorganism has been carried out . C . aceticum was Gram negative . The cells were rodshaped and peritrichously flagellated . Round spores were formed in terminal position . The DNA contained 33 mol % guanine plus cytosine . The organism was obligately anaerobic and grew either chemolithotrophically with H2 + CO2 or chemoorganotrophically with compounds such as fructose, L-glutamate, L-malate or pyruvate . H2 and CO2 were converted to acetic acid according to the following equation: 2CO2 + 4H2 leads to CH3COOH + 2H2O The optimal temperature for growth was 30 degrees C . The optimal pH for chemolithotrophic growth was 8.3 . The doubling times for chemolithotrophic and chemoorganotrophic growth were 25 and 8 h, respectively.

Coll Relat Res, 1981, 1(1), 59 - 72
Production of collagenase inhibitor by the growth cartilage of embryonic chick bone: isolation and partial characterization; Yasui N et al.; Production of collagenase and collagenase inhibitors by the explants of epiphyseal, metaphyseal and diaphyseal regions of embryonic chick limbs during development has been investigated . Collagenase-inhibitory activity was first detected in the culture medium of the diaphyseal region of limbs at stage 36 where cartilage matrix erosion began to occur . However, neither active nor latent collagenase was detected in the media of any regions examined . At stage 38 and later, the maximum production of the inhibitory activity was observed in the explants of metaphyseal region (growth cartilage), while collagenase production was only in the diaphyseal region . Two collagenase inhibitors were isolated and purified approximately 150 fold from the culture medium of stage 38 and 43 metaphyseal regions of limbs by anion- and cation-exchange chromatography followed by gel filtration . The inhibitors are cationic proteins with the same molecular weight of approximately 25,000, but slight difference in molecular charge . They are heat-stable and inhibit collagenases from tadpole skin, chick bone and skin and human granulocytes and gelatinases from human granulocytes and chick skin as well as trypsin, but not Clostridium histolyticum collagenase . A possible function of the inhibitors in multistep regulations of the collagen-degrading enzyme system at the region of osteo-chondral transition is discussed.

J Med Primatol, 1981, 10(4-5), 263 - 4
Acute gastric dilation and rupture in Macaca arctoides associated with Clostridium perfringens; Christie RJ et al.; A clinical case of gastric dilation and rupture is described in an adult male Macaca arctoides . Clostridium perfringens was isolated from the heart blood and liver . Data collected from our macaque colony for 13 years indicated a mortality rate of 3.5% due to acute dilation.

Microbiol Immunol, 1981, 25(9), 915 - 27
Characterization of two inducible bacteriophages, alpha 1 and alpha 2, isolated from Clostridium botulinum type A 190L and their deoxyribonucleic acids; Kinouchi T et al.; Two inducible bacteriophages, alpha 1 and alpha 2, isolated from Clostridium botulinum type A strain 190L and their deoxyribonucleic acids (DNAs) were purified and characterized . Phage alpha 1, which is unable to form plaques on any strain of C . botulinum, was produced in large quantities after treatment with mitomycin C (MC), whereas phage alpha 2, which was induced in much lower quantities than phage alpha 1, propagated in cultures of type A strain Hall . The phage DNAs were exclusively synthesized after induction with MC . Alpha 1 and alpha 2 DNAs had sedimentation coefficients of 34.0 and 30.6 S, corresponding to molecular weights of 31.9 x 10(6) and 23.5 x 10(6), respectively . The buoyant density in CsC1 was 1.682 g/cm3 for alpha 1 DNA and 1.680 g/cm3 for alpha 2 DNA . Based on thermal denaturation characteristics, the genomes of both phages were shown to be double-stranded DNAs . Agarose gel electrophoretic profiles of the phage DNAs digested with restriction endonuclease EcoRI revealed nine fragments for alpha 1 DNA and six fragments for alpha 2 DNA . The molecular weights of the phage DNAs as determined by restriction enzyme analysis were 30.55 x 10(6) for alpha 1 DNA and 25.83 x 10(6) for alpha 2 DNA . Nontoxigenic mutants obtained from strain 190L could, like the toxigenic parent strain, produce the two phages after treatment with MC . Lysogenic conversion to toxigenicity by phage alpha 2 was not observed with the nontoxigenic mutants . It seems likely that there is no relationship between either phage genome and the toxigenicity of C . botulinum type A.

Biochem J, 1981 Jan 1, 193(1), 267 - 73
Effect of bile salts on the hydrolysis of gangliosides, glycoproteins and neuraminyl-lactose by the neuraminidase of Clostridium perfringens; Gatt S et al.; Studies were done on the effect of bile salts on the rates of hydrolysis of the N-acetylneuraminyl linkages of several sialic acid-containing compounds by the neuraminidase of Clostridium perfringens . When GM3-ganglioside, two glycolipids (glycophorin and orosomucoid) and neuraminyl-lactose were used as substrates, hydrolysis was obtained even in the absence of bile salts, but addition of this detergent, below its critical micellar concentration, increased the reaction rates; above the critical micellar concentration of the detergent rates decreased again . When a second ganglioside, GM1, was used as substrate, the requirement for bile salts was absolute; hydrolysis was not observed at all without this detergent . With increasing concentrations of bile salt and in the presence of high concentrations of enzyme, rates of hydrolysis increased, reaching maximal values at fixed ratios of bile salt to GM1-ganglioside . Physical measurements showed that mixtures of bile salt and GM1-ganglioside form mixed micelles that have a higher critical micellar concentration, a lower molecular weight and greater axial ratio than the corresponding micelles of pure GM1-ganglioside.

Zentralbl Chir, 1981, 106(13), 873 - 81
{Epidemiology of nosocomial clostridial myonecrosis (gas gangrene) (author's transl)}; Zavadova M; The occurence of Clostridium perfringens in the environment of different hospital wards and on the skin and in the faeces of in-hospital patients was examined . The author holds the view that most cases of manifest gas gangrene are caused by autogenous infection and not coming from outside.

Cancer Biochem Biophys, 1981, 5(3), 141 - 6
Neuraminidase activity and cell surface sialic acid turnover in Rous sarcoma virus transformed chick embryo fibroblasts; Myers-Robfogel MW et al.; Neuraminidase activity of Rous sarcoma virus transformed chick embryo fibroblasts (RSV-CEF) was assayed using an exogenous substrate, neuraminlactitol-{3H}, and endogenous, cell surface {14C}-N}-acetyl-neuraminic acid . RSV-CEF had higher neuraminidase activity toward both substrates than did chick embryo fibroblasts (CEF) or nontransformed, Rous associated virus infected CEF (RAV-CEF) . The total sialic acid content of RSV-CEF was lower than CEF or RAV-CEF, and more of the total sialic acid was accessible to extracellular Clostridium perfringens neuraminidase . Activity of the enzymes synthesizing and degrading the substrate for sialyltransferase, cytidine-5'-monophosphate-N-acetyl-neuraminic acid (CMP-AcNeu) was measured in order to determine whether control of substrate levels for sialyltransferase might contribute to the decreased levels of glycoprotein bound sialic acid . No change in activity of these enzymes was found in RSV-CEF as compared to CEF or RAV-CEF.

Microbiol Immunol, 1981, 25(1), 1 - 11
Deconjugation of bile salts by Bacteroids and Clostridium; Masuda N; Deconjugation of bile salts by four strains of Bacteroides and four strains of Clostridium was studied by use of resting cells and cell-free culture supernatants . Bacteroids strains yielded active cells but showed relatively low bile salt hydrolase (BSH) activity in the culture supernatants while the reverse was the case for the spore-forming clostridial strains . BSH was formed constitutively and was oxygen insensitive . The optimum pH was between 4.5 and 5.0 . Marked substrate specificity was found in two strains, one Clostridium and one Bacteroides, which showed restricted activity against taurine conjugates . Bacteroides in general attacked the taurine conjugates of dihydroxy bile acids more readily than the trihydroxy taurine conjugates . Deconjugated bile acid moieties were further modified by some resting cells, depending on the bacterial strain while no enzymatic activity other than that of BSH was found in the culture supernatants . Cells of B . fragilis 2536 performed 7 alpha-dehydrogenation when the pH of the medium allowed the reaction, and this oxidative process was markedly enhanced in the presence of an abundant supply of oxygen as a terminal electron acceptor . C . perfringens PB 6K produced the 3- keto product in addition to the 3 beta-hydroxy derivative of the liberated bile acids and the formation of the latter derivative seemed to take place without preliminary deconjugation.

Antimicrob Agents Chemother, 1981 Jan, 19(1), 51 - 5
Susceptibility of anaerobic bacteria to nine antimicrobial agents and demonstration of decreased susceptibility of Clostridium perfringens to penicillin; Marrie TJ et al.; The activity of moxalactam, cefoxitin, cephalothin, cefamandole, chloramphenicol, clindamycin, metronidazole, and ticarcillin was determined against 344 isolates of anaerobic bacteria . The activity of penicillin G was determined as well for 234 isolates not of the Bacteroides fragilis group . Moxalactam was more active than cephalothin and cefamandole and slightly less active than cefoxitin . Metronidazole was the most active antimicrobial agent against the B . fragilis group, whereas chloramphenicol was most active overall . Clostridium species were the most resistant group of organisms tested . Relatively high concentrations of penicillin were required to inhibit the C . perfringens strains: 80% at 0.5 U/ml and 100% at 16 U/ml . Our study demonstrates the need for periodic anaerobe susceptibility testing in order to better guide empiric antibiotic therapy.

Equine Vet J, 1981 Jan, 13(1), 56 - 8
A negative serological relationship between cases of grass sickness in Scotland and Clostridium perfringens type A enterotoxin; Gilmour JS et al.; In an attempt to compare the equine grass sickness as reported in Europe with that described in the Republic of Colombia, sera from horses experiencing grass sickness in Scotland were used in neutralisation tests with Clostridium perfringens type A enterotoxin . The sera, from acute and chronic cases of the disease, failed to neutralise either crude or partially-purified enterotoxin . Neither were precipitin lines formed when the sera were treated against the toxin in immunoelectrophoresis . These results suggest that grass sickness in Europe and the equine disease in Colombia have a different aetiology.

Appl Environ Microbiol, 1981 Jan, 41(1), 315 - 7
Enterotoxin formation by Clostridium perfringens type A in a defined medium; Labbe RG; Enterotoxin was produced by 9 of 10 strains of Clostridium perfringens type A when grown in a defined medium . Additional dextrin increased the amount of enterotoxin in extracts of sporulating cells of strain NCTC 10239.

Appl Environ Microbiol, 1981 Jan, 41(1), 124 - 9
Effects of pH shifts, bile salts, and glucose on sporulation of Clostridium perfringens NCTC 8798; Hickey CS et al.; The sporulation of Clostridium perfringens NCTC 8798 was studied after exposing vegetative cells to: pH values of 1.5 to 8.0 in fluid thioglycolate broth (for 2h) and then transferring them to Duncan-Strong (DS) sporulation medium; sodium cholate or sodium deoxycholate (0.3 to 6.5 mM) in DS medium; or Rhia-Solberg medium with 0.4% (wt/wt) starch, glucose, or both added at 0 to 55 mM . At pH 1.5, no culturable heat-resistant spores were formed . For cells exposed to pH 3.0, 4.0, 5.0, or 6.0, increases in heat-resistant spores were not seen until after a lag of 12 to 13 h, whereas the lag was only 2 to 3 h for cells exposed to pH 7.0 or 8.0 . Maximal spore crops were produced after only 6 to 8 h for cells exposed to pH 7 or 8, but 16 to 18 h was required for production of maximal spore crops by cells exposed to the lower-pH media . The addition of sodium cholate (3.5 to 6.5 mM) to DS medium only slightly reduced the culturable heat-resistant spore count from 1.9 X 10(7) to 3 X 10(6)/ml . The addition of 1.8 mM or more sodium deoxycholate reduced the culturable heat-resistant spore count to less than 10/ ml . When either starch or glucose alone was added to Rhia-Solberg medium there was no production of culturable heat-resistant spores, but a combination of 0.4% (wt/wt) starch and 4.4 mM glucose yielded 6 X 10(5) spores/ml . The spore production remained at this level for glucose concentrations of 6 to 22 mM, but then declined to about 3 X 10(3) spores per ml at higher concentrations.

Zentralbl Bakteriol Mikrobiol Hyg {B}, 1981 Jan, 172(4-5), 427 - 33
Prevalence of food-poisoning (enterotoxigenic) Clostridium perfringens type A in blood and fish meal; Chakrabarty AK et al.; Four of 9 strains of Cl . perfringens type A isolated from 4 of 7 blood meal and 5 of 12 fish meal samples were found to be enterotoxigenic on biological tests and immunodiffusion test with enterotoxin specific serum . Enterotoxin produced by these strains was identical with that of food-poisoning strains . Of these 4 strains, 3 were heat-resistant and 1 was heat-sensitive spore formers . Rate of sporulation of enterotoxigenic strains was greater than the non-enterotoxigenic ones . Animal and public health significance of these strains were discussed.

Microbiol Immunol, 1981, 25(5), 423 - 32
Pharmacological effect of beta toxin of Clostridium perfringens type C on rats; Sakurai J et al.; The biological effect of purified beta toxin of Clostridium perfringens type C in vivo was investigated . After intravenous injection of the purified beta toxin into rats, a rise in blood pressure and a simultaneous fall in heart rate were observed . After the blood pressure reached a maximum, the heart rate recovered gradually, and electrocardiographic and respiratory changes began . The rise in blood pressure induced by beta toxin tended to be proportional to the amount of toxin . The latent period between the injection of toxin and the onset of the increase, and also the time between the injection and the maximum pressure induced by the toxin decreased with increasing concentration of the toxin . A good correlation was found between the factor producing the rise in blood pressure and beta toxin . Alpha adrenergic and ganglionic blocking agents reduced blood pressure levels elevated by beta toxin . The data suggest that the toxin causes a release of catecholamines, and that the increase in blood pressure was induced by released catecholamines.

Ann Anesthesiol Fr, 1981, 22(4), 351 - 8
{Postoperative gas gangrene . Apropos of 22 cases}; Delalande JP et al.; The authors report twenty two cases of post-operative gas gangrene . In the series studied mortality was 40.9 p . 100, independent of age and sex . Rapidly progressive forms were the most severe . The delay before effective treatment was prescribed influenced prognosis . In clinical terms, shock and associated renal insufficiency were grave, as well as a picture of respiratory distress which led, in certain cases, to contra-indication of one of the therapeutic possibilities, i.e . that of hyperbaric oxygen . Responsible organisms could be isolated in nineteen cases from local samples . There was a marked predominance (15 cases) of clostridium perfringens . Contamination with aerobic flora was common . Examination to assess favourizing circumstances led essentially to a conclusion of the role of microbial contamination, ischemia, broad spectrum antibiotics, absence of appropriate antibiotics and underlying immuno-depression . Treatment was based in the majority of cases on the triple combination of antibiotics, surgery and hyperbaric oxygen, as well as the correction of any general systemic disorders . Mortality was markedly reduced (31 p . 100) in patients receiving complete and early treatment . The gravity and recrudescence of disorders due to anaerobic organisms lead the authors to review current therapeutic possibilities . Appropriate treatment should be prescribed in all situations where an infection due to anaerobic organisms is feared, and should cover the risk of clostridial infection (penicillin 200,000 mu/kg/24 h) as well as the risk of bacteroides (metronidazole 25 mg/kg/24 h) . Curative treatment should be prescribed, even in the absence of bacteriological proof, on the basis of presumptive clinical evidence, this being a true emergency which should not be delayed under any circumstances.

J Biol Chem, 1980 Dec 25, 255(24), 11737 - 42
Purification and properties of alpha-N-acetylgalactosaminidase from Clostridium perfringens; Levy GN et al.; Exo-alpha-N-acetylgalactosaminidase has been purified 8000-fold from Clostridium perfringens by gel filtration, ion exchange chromatography, isoelectric precipitation, and negative adsorption on human O type erythrocytes . The resulting enzyme is active at physiological pH and temperature . Phenyl glycosides, oligosaccharides, mucins, glycolipids, and cell membranes are substrates for this enzyme . The result of enzyme action on blood type A erythrocytes is the loss of A activity and the simultaneous appearance of H activity, characteristic of the O blood group type . Polyacrylamide gel electrophoresis in sodium dodecyl sulfate demonstrates electrophoresis in sodium dodecyl sulfate demonstrates that the blood group A-destroying activity is distinct from the other glycosidase activities found in C . perfringens.

Appl Environ Microbiol, 1980 Dec, 40(6), 1007 - 11
Plasmid-controlled mercury biotransformation by Clostridium cochlearium T-2; Pan-Hou HS et al.; A strain of Clostridium cochlearium having methylmercury-decomposing ability was isolated . The ability was cured by the treatment with acridine dye and recovered by the conjugation of the cured strain with the parent strain . The cured strain then showed the activity to methylate mercuric ion as previously reported (M . Yamada and K . Tonomura, J . Ferment . Technol . 50:159-166, 1971) . These results and the agarose gel electrophoretic pattern of the deoxyribonucleic acids from the lysates indicate a possible role of plasmids in controlling the mercury biotransformation of the two opposite directions in a single bacterial strain: methylation in the absence of the plasmid and demethylation in the presence of it . A possible mechanism for mercury resistance involving hydrogen sulfide is discussed.

South Med J, 1980 Dec, 73(12), 1644 - 5
Clostridium paraputrificum sepsis in sickle cell anemia; Brook I et al.; We have described clostridial sepsis in a child who presented with an abdominal crisis of sickle cell disease . Clostridium paraputrificum should be included in the list of organisms known to cause sepsis in patients with sickle cell anemia.

Onderstepoort J Vet Res, 1980 Dec, 47(4), 287 - 9
Effective immunization of lambs against enterotoxaemia; Cameron CM; In contrast to adult sheep, 2- to 3-month-old lambs do not respond well to a single injection of Clostridium perfringens Type D oil adjuvant epsilon toxoid . This unresponsiveness can be overcome, however, by administering 2 injections of oil adjuvant vaccine or one injection of oil adjuvant followed 4 weeks later by an injection of alum-precipitated toxoid . The latter procedure evokes protective antitoxin levels which persist for 8 months, and a booster injection of alum-precipitated toxoid given at this stage results in an immunity which lasts for at least 1 year.

Appl Environ Microbiol, 1980 Dec, 40(6), 1023 - 6
Effect of fermentation conditions on toxin production by Clostridium botulinum type B; Siegel LS et al.; To obtain high yields of toxin for the preparation of purified neurotoxoids, we examined the time of appearance and the quantity of toxin produced by the Bean strain of Clostridium botulinum type B under various conditions by using a fermentor system . The medium employed consisted of 2.0% casein hydrolylsate and 1.5% yeast extract plus an appropriate concentration of glucose . The maximum toxin concentration (4 x 10(5) to 5 x 10(5) mouse median lethal doses per ml) was attained within 48 h under the following fermentation conditions: an initial glucose concentration of 0.5 or 1.0%, a temperature of 35 degrees C, a nitrogen overlay at a rate of 5 liters/min, and an agitation rate of 50 rpm.

Pediatrics, 1980 Dec, 66(6), 936 - 42
Infant botulism: clinical spectrum and epidemiology; Thompson JA et al.; Between 1977 and 1979, 12 cases of infant botulism were diagnosed in Utah, and 87 control patients (normal, nonbotulism neurologic disease, and nonbotulism systemic disease) were evaluated . Observations from these patients suggest an expanded clinical spectrum of infant botulism including asymptomatic carriers of organism; mild hypotonia and failure to thrive; typical cases with constipation, bulbar weakness, and hypotonia; and children with a picture compatible with sudden infant death syndrome . Clostridium botulinum was isolated from the stools of three normal control infants and nine control infants who had neurologic diseases that were clearly not infant botulism . These infants were termed "asymptomatic carriers" of the organism . The occurrence of the asymptomatic carrier state suggests that a diagnosis of infant botulism cannot be made on a basis of culture results alone, but must rest in historical documentation and physical confirmation of progressive bulbar and extremity weakness with ultimate complete resolution of symptoms and findings over a period of several months . A common set of environmental features characterizes the home environment of children with infant botulism and "asymptomatic carriers" and includes: nearby constructional or agricultural soil disruption, dusty and windy conditions, a high water table, and alkaline soil conditions.

Radiology, 1980 Dec, 137(3), 625 - 7
Clostridium septicum infection associated with colonic carcinoma and hematologic abnormality; Schaaf RE et al.; Six patients with Clostridium septicum sepsis seen at Duke University Medical Center over a two-year period also had other abnormalities, consisting of hematologic disorders in 3 and colon tumors in 3 . Three patients died of sepsis; 2 survived following disarticulation of the arm to control gas gangrene, while the sixth patient survived the sepsis but died of metastatic disease . When anaerobic cultures are positive for C . septicum, antibiotics should be given immediately . The high incidence of underlying colon tumor, especially in the cecum, should prompt consideration of a barium-enema examination.

Pediatrics, 1980 Dec, 66(6), 928 - 31
Neonatal necrotizing enterocolitis associated with penicillin-resistant, toxigenic Clostridium butyricum; Sturm R et al.; The cause of neonatal necrotizing enterocolitis (NEC) is unknown . An association between NEC and clostridial infection has been reported from several centers, but the organisms have not been extensively characterized . Clostridium butyricum was isolated from the peritoneal fluid and cerebrospinal fluid of a neonate with NEC . The organism was resistant to the penicillins, but sensitive to vancomycin . Toxin production was demonstrated . Although the role of clostridial toxins in the pathogenesis of NEC is unknown, clostridial toxins are well established as the causes of two other intestinal diseases (antibiotic-associated pseudomembranous colitis and pig-bel) . Further investigation of the role of clostridia in the pathogenesis of NEC and of the use of oral, nonabsorbable antibiotics in the treatment of NEC is needed.

Infect Immun, 1980 Dec, 30(3), 668 - 73
Purification and characterization of two components of botulinum C2 toxin; Ohishi I et al.; Two dissimilar proteins, designated as components I and II, of botulinum C2 toxin elaborated by strain 92-13 were purified to a homogeneous state . The molecular weights determined by sodium dodecyl sulfate gel electrophoresis were 55,000 for component I and 105,000 for component II . Whereas each component showed no or feeble toxicity even after being treated with trypsin, the toxicity was elicited when these two components were mixed and trypsinized . The toxicity of the mixture of components I and II at a ratio of 1:2.5 on a protein basis was 2.2 X 10(4) mouse intraperitoneal 50% lethal doses per mg of protein and increased by 2,000 times or more when treated with trypsin . These results indicate that the molecular characteristics of botulinum C2 toxin differ from those of the toxin of Clostridium botulinum types A through F in that C2 toxin is constructed with two separate protein components, which are not covalently held together, and that its toxicity is elicited by cooperation of the two components.

Infect Immun, 1980 Dec, 30(3), 656 - 60
Antigenic similarity of toxins produced by Clostridium botulinum type C and D strains; Oguma K et al.; Antisera against purified type C1 toxin of Clostridium botulinum and its heavy-chain component cross-neutralized type D toxin . Antisera against partially purified type D toxin cross-neutralized type C1 toxin . From the latter serum, a component which neutralized only type D toxin and a component which equally neutralized both C1 and D toxins were obtained . We concluded that the cross-neutralization was not due to the fact that type C and D strains produce both C1 and D toxins but rather to the fact that the toxins have an antigen(s) common to their molecules . The results of the agar gel-double-diffusion test also supported this conclusion.

Int J Zoonoses, 1980 Dec, 7(2), 78 - 84
Active perfringocin typing of food poisoning strains of Clostridium perfringens type A--a new tool for epidemiological investigations; Satija KC et al.; Active perfringocin typing, as an epidemiological tool, for investigation into food poisoning outbreaks due to C . perfringens type A has been developed . Ninety strains included in this study were from England, America, India, Japan, France and Canada Nine indicator strains exhibiting different patterns of inhibition were selected for perfringocin typing . With these indicator strains, 81.11 per cent of the strains were typable into ten perfringocin types . Type "a" and "b" alone could type 47.92 per cent of the strains . Type "f" was predominantly found to be for the American strains and "b" and "c" for the Indian strains . Active perfringocin typing developed for the first time for the food poisoning strains of C . perfringens type A was expected to prove useful in the epidemiological investigations.

J Infect Dis, 1980 Dec, 142(6), 899 - 902
Passive bacteriocin typing of strains of Clostridium perfringens type A causing food poisoning for epidemiologic studies; Satija KC et al.; A passive bacteriocin typing system was developed for use as an epidemiologic tool to study outbreaks of food poisoning caused by Clostridium perfringens type A . The 90 strains tested were from England, the United States, India, Japan, France, and Canada . Forty of 74 strains produced bacteriocin when irradiated with ultraviolet light for 40-90 sec . Eight bacteriocins were concentrated with ammonium sulfate and titrated on a common indicator strain . These bacteriocins were found to be specific for C . perfringens type A . All of the 90 strains were typable, with 16 distinct bacteriocin types . Most (85.56%) of the strains were types 1-6 . Strains of C . perfringens type A clustered into certain patterns of bacteriocin types according to their country of origin and source of isolation.

Appl Environ Microbiol, 1980 Dec, 40(6), 994 - 1002
Behavior of pathogenic bacteria in the oyster, Crassostrea commercialis, during depuration, re-laying, and storage; Son NT et al.; Oysters (Crassostrea commercials) harvested from major cultivation areas within the state of New South Wales, Australia, were commonly contaminated with low levels of the food-poisoning organisms Bacillus cereus, Clostridium perfringens, and Vibrio parahaemolyticus . Salmonella was found in oysters on only one occasion . These bacteria were cleansed from oysters during oyster purification by re-laying in a non-polluted waterway . Oysters were laboratory contaminated to levels in excess 1,000 cells per g with either B . cereus, C . perfringens, V . parahaemolyticus, Salmonella typhimurium, or S . senftenberg . These species were cleansed from such oysters during purification in a laboratory depuration unit that used ultraviolet light for sterilizing the depuration water . Escherichia coli was also cleansed from oysters under the same re-laying or depuration conditions so that its measurement alone could be used to indicate the cleansing of the above pathogenic species . The levels of these bacteria were also measured during the storage of oysters under conditions that occur during marketing . While B . cereus counts remained relatively stable during storage, the Salmonella spp . gradually decreased in numbers and C . perfringens rapidly died off . V . parahaemolyticus counts increased slightly during the first 4 days of storage, after which decreases occurred.

J Clin Pathol, 1980 Dec, 33(12), 1189 - 92
Detection of Bacteroides fragilis and Bacteroides melaninogenicus by direct immunofluorescence; Labbe M et al.; A new diagnostic kit, which contains a polyvalent antiserum for either Bacteroides fragilis or Bacteroides melaninogenicus, was tested for reliability and specificity on 146 clinical samples of different origin . A correlation between the culture and immunofluorescence was observed for B . fragilis in 87.39% of cases and for B . melaninogenicus in 81.48% of cases . When pure cultures were tested, aerobically as well as anaerobically, false-positive reactions were observed with staphylococci and Clostridium ramosum spores . The well-defined morphology of these bacteria and spores allows for the elimination of any diagnostic error . The method is rapid, and the margin for error is limited . The test gives a semiquantitative idea of the number of bacteroides organisms present in the clinical specimens even in the presence of a mixed flora.

J Biol Chem, 1980 Nov 25, 255(22), 10644 - 50
Regulation of phosphatidylcholine biosynthesis in cultured chick embryonic muscle treated with phospholipase C; Sleight R et al.; Cultures of embryonic chick muscle cells grown in medium containing phospholipase C from Clostridium perfringens incorporated {3H}choline into lipid at a rate 3- to 5-fold higher than control cultures . To determine the mechanism by which stimulation of phosphatidylcholine synthesis occurred in phospholipase C-treated cells, activities of enzymes and levels of intermediates in the biosynthetic pathway for phosphatidylcholine were examined . Activities of choline kinase, choline phosphotransferase, glycerol-3-phosphate dehydrogenase, glycerol-3-phosphate acyltransferase, acylglycerol-3-phosphate acyltransferase, and phosphatidic acid phosphatase in phospholipase C-treated cells were the same or only slightly higher than in control cells . CTP:phosphocholine cytidylyltransferase, on the other hand, was 3 times as active in homogenates from phospholipase C-treated cells . Levels of phosphocholine decreased and levels of CDP-choline increased in phospholipase C-treated cells, and a calculation of the disequilibrium ratio indicated that the cytidylyltransferase reaction was not at equilibrium . The cytidylyltransferase was, thus, identified as the regulatory enzyme for choline flux in these cells . The cytidylyltransferase was located in both the cytosolic and particulate fractions from cultured muscle cells and a much larger portion of enzyme activity was associated with the particulate fraction in cells treated with phospholipase C . Sonicated preparations of total chick lipids, phosphatidylethanolamine, and phosphatidylserine greatly stimulated the cytosolic cytidylyltransferase activity but had no effect on the particulate enzyme . Neither stimulation of incorporation of {3H}choline into lipid nor activation of the cytidylyltransferase was dependent on protein synthesis . A model for the mechanism of regulation of phosphatidylcholine synthesis in embryonic chick muscle is presented.

Biochim Biophys Acta, 1980 Nov 20, 626(1), 234 - 43
Purification and composition of colonic epithelial mucin; LaMont JT et al.; Colonic mucin was purified from homogenized scrapings of rat colonic epithelial cells using gel filtration and ion-exchange chromatography . High molecular weight water-soluble mucin was separated from low molecular weight proteins by gel exclusion chromatography on Sepharose 4B, and was further separated into two major mucin fractions and several non-mucin fractions on DEAE-cellulose . Fraction IV, the major mucin, was a sulphated glycoprotein with 62% carbohydrate by weight, and high concentrations of serine and threonine . A more acidic mucin, fraction V, had similar composition . Approx . 85% of the sialic acid of fractions IV and V were removed after incubation with Clostridium perfringens neuraminidase . Blood group A but not group H activity was present in fractions III, IV, and V . Ultracentrifugation experiments showed that fraction IV migrated as a single peak, whereas fraction V contained two components . Our study indicates that colonic mucin consists of at least two closely related acidic high molecular weight glycoproteins which can be separated from non-mucin contaminants by ion-exchange chromatography.

Sem Hop, 1980 Nov 18-25, 56(43-44), 1793 - 5
{Susceptibility of anaerobic bacteria to cefoxitin (author's transl)}; Freydiere AM et al.; Nine strains of Clostridium perfringens type A and 10 strains of Bacteroides fragilis s . spec . fragilis, recently isolated from blood culture, were tested for their cefoxitin susceptibility . The MIC for Clostridium perfringens ranged from 0,6 to 5 mg/l and 1,2 to 10 mg/l for Bacteroides fragilis . In most cases, the MBC were identical with the MIC for Clostridium perfringens, but for Bact . frag . there was a difference of 2 dilutions in their MBC . Cefoxitin activity was not modified by a variation in inoculum density.

Biochem J, 1980 Nov 15, 192(2), 665 - 72
Efficiency of ferredoxins and flavodoxins as mediators in systems for hydrogen evolution; Fitzgerald MP et al.; 1 . The efficiencies of ferredoxins and flavodoxins from a range of sources as mediators in systems for hydrogen evolution were assessed . 2 . In supporting electron transfer from dithionite to hydrogenase of the bacterium Clostridium pasteurianum, highest activity was shown by the ferredoxin from the cyanobacterium Chlorogloeopsis fritschii and flavodoxin from the bacterium Megasphaera elsdenii . The latter was some twenty times as active as comparable concentrations of Methyl Viologen . Ferredoxins from the cyanobacterium Anacystis nidulans and the red alga Porphyra umbilicalis also showed high activity . 3 . In mediating electron transfer from chloroplast membranes to Clostridium pasteurianum hydrogenase the flavodoxin from Anacystis nidulans proved the most active with Nostoc strain MAC flavodoxin and Porphyra umbilicalis ferredoxin also being appreciably more active than other cyanobacterial and higher plant ferredoxins . 4 . In both hydrogenase systems the ferredoxin and flavodoxin from the red alga Chondrus crispus and the ferredoxin from another red alga Gigartina stellata showed very low activity . 5 . There appeared to be no apparent correlation of efficiency in supporting hydrogenase activity with midpoint redox potential (Em) of the mediators, though some correlation of Em with the efficiency of the mediators in supporting NADP+ photoreduction by chloroplasts, or pyruvate oxidation by a Clostridium pasteurianum system, was evident . 6 . Activity of the mediators in the hydrogenase systems therefore primarily reflects differences in tertiary structure conferring differing affinities for the other components of the systems.

J Biol Chem, 1980 Nov 10, 255(21), 10460 - 3
Chemical cross-linking stabilizes the enzymic activity and quaternary structure of formyltetrahydrofolate synthetase; de Renobales M et al.; Clostridium cylindrosporum formyltetrahydrofolate synthetase tetramers cross-linked with dimethyl suberimidate remained active in the absence of the monovalent cations normally required for enzymic activity and the tetrameric conformation . The modified enzyme was analyzed by sodium dodecyl sulfate electrophoresis, sedimentation velocity, and gel permeation chromatography . Under the experimental conditions used, the enzyme was only partially cross-linked; 74% of the enzyme was cross-linked dimer or monomer . Nonetheless, the modified enzyme is able to retain enzymic activity and the tetrameric structure under conditions where native enzyme would be completely dissociated and inactivated . The result suggests that cross-linked dimers strongly associate with each other and with monomers . Flame emission spectroscopy indicates that cross-linked enzyme contains two monovalent cations per tetramer.

Vet Rec, 1980 Nov 8, 107(19), 445 - 6
An outbreak of type C botulism in captive monkeys; Smart JL et al.; In an outbreak of type C botulism in a group of captive primates, six squirrel monkeys, six white throated capuchin monkeys and two weeper capuchin monkeys succumbed rapidly and died . Clostridium botulinum type C toxin was detected in the remains of the chopped chicken feed and in nine of 11 blood samples and one of three stomach contents samples from the affected animals.

Am J Dis Child, 1980 Nov, 134(11), 1052 - 6
Anaerobic bacteremia in children; Brook I et al.; Twenty-nine anaerobic isolates were recovered from 28 pediatric patients with anaerobic bacteremia: 14 Bacteroides sp (11 in the B fragilis group); four anaerobic Gram-positive cocci; four Clostridium sp; four Propionibacterium acnes; and three Fusobacterium sp . No aerobic bacteria were isolated from these patients . The gastrointestinal tract was the possible portal of entry in 13 instances, eight of which were due to Bacteroides organisms, four to Clostridium sp, and one to F nucleatum . The ear, sinus, and oropharynx were probable portals of entry in seven instances, four of which were due to Peptococcus sp and two to Fusobacterium sp . Five patients (18%) died; four had bacteremia caused by B fragilis group, and one had bacteremia caused by P acnes . The average duration of antimicrobial therapy was 20 days (range, seven to 72 days) . The early recognition and rapid institution of appropriate antimicrobial and surgical therapy are of utmost importance in improving the outcome of these patients.

Am J Clin Nutr, 1980 Nov, 33(11 Suppl), 2527 - 32
Studies on the epidemiology of antibiotic-associated Clostridium difficile colitis; Fekety R et al.; Vancomycin protects hamsters from the development of Clostridium difficile colitis after treatment with clindamycin, and vancomycin is useful in treatment of humans with the disease . Relapses have occurred in both hamsters and humans when vancomycin is discontinued . Vancomycin appears to enhance susceptibility to colonization with C . difficile by eliminating competing intestinal organisms . The nature of these organisms is not known, but various tools are now available to aid in identifying them . Cancer chemotherapeutic agents should be added to the list of factors such as surgery and antibiotics that may predispose to emergence of C . difficile . The number of organisms required for colonization of antibiotic-treated hamsters is low and cross-infection seems to play a role in the disease in hamster colonies . The organism can be detected on surfaces in rooms of patients with the disease, and on the hands of personnel caring for them . Outbreaks of the disease have been recognized . Our results suggest isolation precautions should be used to prevent spread of the organism from patients with the disease to others being treated with antibiotics.

J Med Microbiol, 1980 Nov, 13(4), 609 - 12
Effect of temperature, humidity and exposure to oxygen on the survival of anaerobic bacteria; Hoffmann S et al.; The relative effect of humidity, temperature and exposure to oxygen on the survival of 11 strains of Bacteriodes spp., Fusobacterium necrophorum, Eubacterium lentum, Clostridium ramosum, Peptococcus asaccharolyticus and Peptostreptococcus anaerobius was evaluated . Suspensions of the test strains were applied to membrane filters either in empty sterile petri dishes or on plates of non-nutrient agar to provide dry or moist conditions . The filters were stored aerobically or anaerobically in an atmosphere containing 10% CO2 at either 4 degrees C or 35 degrees C . After holding periods of 0.375-384 h, the filters were transferred to nutrient plates inside an anaerobic glove box . After incubation the growth on the filters was compared with that on control filters that had not been stored . Survival was better at 4 degrees C than at 35 degrees C . For the gram-negative organisms, survival was better under moist than under dry conditions, whereas for gram-positive organisms this finding was sometimes reversed . The results indicate that, generally, anaerobic bacteria kept under aerobic conditions survive best under moist conditions at 4 degrees C.

Gastroenterology, 1980 Nov, 79(5 Pt 1), 948 - 51
Role of Clostridium difficile in a case of nonantibiotic-associated pseudomembranous colitis; Peikin SR et al.; A 57-yr-old woman with chronic diarrhea, mild azotemia, and red cell casts in her urine was found to have pseudomembranous colitis . She had not received antimicrobial agents for at least 2 yr . Membranoproliferative glomerulonephritis was found in kidney biopsy, and her renal function improved spontaneously . Pseudomembranous colitis was diagnosed by endoscopic biopsy . Her stool contained a cytophatic toxin that was neutralized by Clostridium sordellii antitoxin, and Clostridium difficile was cultured on selective media . This case indicates that C . difficile may be a cause of nonantibiotic-associated pseudomembranous colitis, as well as antibiotic-associated pseudomembranous colitis.

Can J Microbiol, 1980 Nov, 26(11), 1275 - 83
Dinitrogen-fixing bacteria: computer-assisted identification of soil isolates; Rennie RJ; Dinitrogen-fixing (acetylene-reducing) bacteria may be readily isolated from soils but extensive biochemical or immunobiological testing, or both, are required to identify them absolutely . A computer-assisted scheme for identification of nine genera of dinitrogen-fixing bacteria was developed and tested . The computer program is based on interpretation of the 70 biochemical tests of the API 20E and 50E, supplemented with tests for acetylene reduction, nitrate and nitrite reduction, catalase, oxidase, motility, and growth on MacConkey's bile salt medium . Dinitrogen-fixing Enterobacteriaceae (Klebsiella pneumoniae, Enterobacter cloacae, and Erwinia herbicola) were accurately identified using the data base in the API analytical profile index . Nonenteric dinitrogen-fixing bacteria (Azotobacter spp., Azospirillum spp., Derxia sp., Rhodospirillum sp., Clostridium sp., and Bacillus spp.) were subjected to these tests to form a new data base for these bacteria . The API tests agreed with standard biochemical tests commonly used to identify these bacteria, were reproducible with time, and were sufficiently unique to permit accurate identification of each species . The use of the API 20E and 50E tests plus the additional seven tests with these known data bases permitted rapid and precise identification of acetylene reducing bacteria from various agricultural ecosystems.

Zh Mikrobiol Epidemiol Immunobiol, 1980 Nov, (11), 42 - 6
{Isolation and properties of a highly purified Clostridium botulinum type F toxin}; Uvarova RN et al.; A scheme for isolation of highly purified type F Cl . botulinum toxin has been developed . The scheme allows one to obtain the toxin with specific activity of 1.5--4.0 X 10(7) Dlm/mg protein, molecular weight of 150,000 daltons, a typical protein UV spectrum and the optical density ratio E280/E260 = 1.98.

J Lipid Res, 1980 Nov, 21(8), 1022 - 31
The enzymes of phospholipid synthesis in Clostridium butyricum; Silber P et al.; We have examined extracts of Clostridium butyricum for several enzymes of phospholipid synthesis . Membrane particles were shown to catalyze the formation of CDP-diglyceride from {3H}CTP and phosphatidic acid . The reaction was dependent on Mg2+ and stimulated by monovalent cations . CDP-diglyceride formed in vitro was found to be a substrate for both phosphatidylglycerophosphate synthetase and phosphatidylserine synthetase . The formation of phosphatidylglycerophosphate from added CDP-diglyceride and {U-14C}sn-glycerol-3-phosphate was dependent on Mg2+ and Triton X-100 . The dephosphorylation of endogenously-generated phosphatidylglycerophosphate to yield phosphatidylglycerol was observed to be pH-dependent . The formation of phosphatidylserine from CDP-diglyceride and L-{3-14C}serine was stimulated by Mg2+ and Triton X-100 . dCDP-diglyceride was a suitable substrate for both phosphatidylglycerophosphate synthetase and phosphatidylserine synthetase . Phosphatidylserine decarboxylase activity was barely detectable in membrane particles from C . butyricum . The addition of E . coli membrane particles provided efficient phosphatidylserine decarboxylase activity in this system . Although plasmalogens are the principal lipids of C . butyricum, none of the products of phospholipid synthesis formed in vitro contained measurable amounts of plasmalogens . The subcellular distribution of both phosphatidylglycerophosphate synthetase and phosphatidylserine synthetase in C . butyricum was also studied . Both were found to be membrane-associated.

Vet Pathol, 1980 Nov, 17(6), 738 - 47
The effects of Clostridium perfringens type A enterotoxin in Shetland ponies--clinical, morphologic and clinicopathologic changes; Ochoa R et al.; Severe abdominal pain, classic colic signs and hemorrhagic gastro-entero-cecocolitis were induced in three conventional Shetland ponies by intravenous injection with Clostridium perfringens Type A enterotoxin . Histological examination showed marked congestion, edema and hemorrhage of the large and small intestine and sloughing of the tips of the intestinal villi . Marked vacuolar degeneration of hepatocytes with dilatation of the spaces of Disse also was found . Clinical changes consisted of severe hypoglycemia, markedly increased aspartate aminotransferase levels and leukopenia that occurred rapidly.

Zh Mikrobiol Epidemiol Immunobiol, 1980 Nov, (11), 60 - 4
{Use of the aggregate hemagglutination method for determining cholera vibrio neuraminidase}; Vertiev IuV et al.; The suitability of the aggregate hemagglutination test for neuraminidase determination in biological fluids has been evaluated . This test allows one to determine neuraminidase at a concentration of 2--4 mg/ml . The sensitivity of this test is more than 2 orders higher than that of gel diffusion technique and the thiobarbituric assay . The test has revealed that neuraminidase produced by Vibrio cholerae is not immunologically related to other kinds of neuraminidase produced by Corynebacterium diphtheriae, Clostridium perfringes and Erysepelothrix insidiosa.

J Med Microbiol, 1980 Nov, 13(4), 573 - 9
The carbon dioxide requirements of anaerobic bacteria; Reilly S; Some clinically significant anaerobic bacteria were incubated in pure culture in anaerobic jars containing a range of atmospheric concentrations of CO2 . Growth of Clostridium perfringens, C . sporogenes and C . septicum was independent of the amount of CO2 . Small supplements of CO2 (0.25%) allowed good growth of the majority of anaerobes studied--an observation contrary to established teaching . The exceptions were Fusobacterium necrophorum which showed an absolute requirement for CO2 of at least 1% and B . melaninogenicus which needed an atmospheric content of 10--40% CO2 for optimal growth . The inclusion of CO2 in the anaerobic jar at a final concentration of 10% is to be recommended for all routine isolation procedures.

Biochemistry, 1980 Oct 14, 19(21), 4801 - 7
Binding of Clostridium perfringens {125I}enterotoxin to rabbit intestinal cells; McDonel JL; 125I-Labeled enterotoxin from Clostridium perfringens was utilized to characterize the association of the enterotoxin with cells isolated from rabbit intestine and tissue homogenates from liver, kidney, and brain . The enterotoxin was found to bind in a specific and saturable manner to cells from intestine and to tissue homogenates from liver and kidney but not the brain . Detailed studies of the binding were carried out with the ileal epithelial intestinal cells . The rate and amount of binding of enterotoxin to cells appeared to be temperature dependent . Apparent affinity and association and dissociation rate constants were calculated for what appeared to be two classes of saturable binding sites . The amount of enterotoxin molecules that bound per milligram of cell protein was similar in tissue of intestinal, liver, and kidney origin (approximately 10(13) molecules/mg of cell protein) . Spontaneous dissociation into the supernatant medium was observed to be much slower than expected from calculations based on the rate of association . Chaotropic ions did not enhance dissociation of the enterotoxin from cells . Enterotoxin binding was demonstrated to be heat labile (binding ability was lost after the enterotoxin was heated for 10 min at 60 degrees C) . A mechanism is described whereby the enterotoxin binds and then is inserted into the membrane where it becomes trapped.

J Clin Pathol, 1980 Oct, 33(10), 1002 - 5
Antimicrobial susceptibilities of Clostridium difficile; Shuttleworth R et al.; The antimicrobial susceptibilities of 78 strains of Clostridium difficile isolated from patients with and without gastrointestinal symptoms were determined and compared . Strains from patients with symptoms were more likely to show resistance to antibiotics . The antimicrobial susceptibilities of toxigenic and non-toxigenic strains were found to be similar.

Atherosclerosis, 1980 Oct, 37(2), 277 - 84
Effect of sialic acid removal on human low density lipoprotein catabolism in vivo; Malmendier CL et al.; This study was undertaken to determine whether sialic acid removal alters the catabolism of low density lipoprotein in humans . Human low density lipoproteins labeled in vitro with 125I were incubated in the presence (termed desialylated) or absence (sialylated) of neuraminidase . The treatment with neuraminidase from Clostridium perfringens removed 90% of the sialic acid residues which did not change the chemical composition of the lipoproteins . Sialylated or desialylated LDL were injected intravenously into normal human subjects . The mathematical analysis of the plasma radioactivity decay curves (followed for 220 h) of desialylated low density lipoproteins, when compared with sialylated LDL, showed a shorter mean transit time (51 h vs 60 h), a 52% faster metabolic catabolic rate and an increased volume of distribution . The data are consistent with a proposed metabolism of low density lipoproteins: in humans, desialylation appears to accelerate the first step of the low density lipoprotein conversion but not to alter its final catabolism.

Obstet Gynecol, 1980 Oct, 56(4), 518 - 21
Clostridium perfringens septicemia following cesarean section; Mariona FG et al.; A Clostridium perfringens infection is described in an 18-year-old primigravida following cesarean section at 43 weeks' gestation . Despite the ubiquitous distribution of these organisms, such infections are rarely encountered in obstetrics . Characteristics of these bacteria and appropriate diagnostic and therapeutic procedures are presented . Despite the infrequency of this potentially lethal complication, survival depends on early diagnosis and aggressive treatment.

Jpn J Med Sci Biol, 1980 Oct, 33(5), 255 - 61
In vitro inactivation of Clostridium botulinum toxins types B, C and E by digestive juices of man and ducks; Notermans S et al.; Inactivation of botulinum toxins type B-L, B-M, C-L and E in the digestive juices of man and ducks was determined . Botulinum toxins of the M size (B-M and E) lost their toxicity completely in the gastric juices of both man and ducks, but toxins of the L size (B-L and C-L) lost their toxicity only partially . The toxins were hardly affected by duodenal fluid even after exposure to gastric juice . The toxins were not inactivated by cecal fluid from ducks . Since human digestive juices and those from ducks appeared to have comparable effects on the inactivation of toxin, differences in susceptibilities of the animal species to orally acquired toxins are in all probability not caused by digestive action of those species.

J Hyg (Lond), 1980 Oct, 85(2), 271 - 4
Clostridium botulinum in British soil; Smith GR et al.; Soil samples from various parts of Britain were examined for Clostridium botulinum by a sensitive technique comparable with that recently used for mud samples from British aquatic environments . The results showed beyond doubt that in Britain the prevalence of the organism in soil is much lower than in mud . Of 174 samples from all sites examined only 10 (5 . 7%) could be shown to contain Cl . botulinum; this finding was consistent with the results of surveys made by less sensitive techniques in 1922, 1928 and 1942 . No type other than B was found . The evidence suggested that in certain localized areas the prevalence was likely to be high . Three sites associated for many years with animals were included in the survey; at the Zoological Society's premises at Regent's Park and Whipsnade, and at the Market paddocks, Gorgie, Edinburgh, the prevalence of Cl . botulinum was either very low, or nil . In an earlier survey of the redeveloped site of the former Metropolitan Cattle Market, London, 25% of soil samples gave a positive result and no less than four types (B, C, D . and E) were demonstrated.

Dig Dis Sci, 1980 Oct, 25(10), 783 - 4
Bacitracin therapy in antibiotic-associated pseudomembranous colitis; Tedesco FJ; Two patients with antibiotic-associated pseudomembranous colitis, and stool positive for Clostridium difficile cytotoxin were successfully treated with oral bacitracin . One patient had previously suffered two relapses of pseudomembranous colitis following successful treatment with vancomycin and one patient was allergic to vancomycin . Bacitracin appears to be a reasonable choice to treat patients with antibiotic-associated colitis who are allergic to vancomycin . Further studies comparing vancomycin and bacitracin are needed.

Lab Anim, 1980 Oct, 14(4), 347 - 51
Enterotoxaemia involving Clostridium perfringens iota toxin in a hysterectomy-derived rabbit colony; Eaton P et al.; During an explosive outbreak of fatal enteropathic disease involving Clostridium perfringens iota (i) toxin, a total of 183 deaths occurred in 18 weeks . The clinical signs and post-mortem findings are reported . Examinations for virus, Bacillus piliformis and coccidia were negative . Clostridium perfringens i toxin was detected in 22 of 27 animals examined (81.5%), but clostridia were not isolated . Various treatments were attempted . It is concluded that i toxin and the syndrome described are closely related.

Antimicrob Agents Chemother, 1980 Oct, 18(4), 566 - 73
Relationship between metronidazole metabolism and bactericidal activity; Chrystal EJ et al.; It has been suggested that the microbicidal effect of metronidazole is mediated by an intermediate in nitro group reduction . We have found that the addition of Escherichia coli enhances the lethal effect of metronidazole on Bacillus fragilis and suggest that this intermediate may form in one bacteria and kill another . Because acetamide forms during the reduction of metronidazole, we examined the possibility that the same partially reduced intermediate in metronidazole reduction may be both an intermediate in the formation of acetamide and the ultimate reactive form of metronidazole which is responsible for its bactericidal action . Thus, we determined the relationship between bacterial survival and the formation of acetamide when cultures of B . fragilis, Clostridium perfringens, and E . coli were incubated anaerobically in the presence of metronidazole . We found that the log of the early bacterial survival was proportional to the formation of acetamide . The rate of loss of metronidazole was not dependent on the concentration of bacteria in the medium, suggesting that any proposed intermediate formed at a rate which was proportional only to the concentration of metronidazole.

J Nutr, 1980 Oct, 110(10), 2103 - 8
Incidence of increased numbers of Clostridium perfringens in the intestinal tract of rats fed xylitol; Wekell MM et al.; Sprague-Dawley rats were fed diets containing up to 20% xylitol for 49 days . When the rats were fed a xylitol regimen intended to produce adaptation to xylitol, approximately half of the animals adapted to xylitol and remained free from diarrhea during the feeding regimen . The other half did not adapt to xylitol and developed severe and persistent diarrhea accompanied by large volumes of intestinal gas . These non-adapted rats had significantly higher levels of intestinal tract Clostridium perfringens (10(6)--10(11) organisms per gram intestinal contents) than did control rats fed a xylitol-free cornstarch diet (0-10(4) organisms per gram) . Rats adapted to dietary xylitol did not have detectable levels of C . perfringens in the gastrointestinal tract.

J Clin Microbiol, 1980 Oct, 12(4), 617 - 9
New presumptive identification test for Clostridium perfringens: reverse CAMP test; Hansen MV et al.; The reverse CAMP test proved to be a highly sensitive test since 97.0% of all Clostridium perfringens cultures tested gave an easily discernible positive reaction.

J Bacteriol, 1980 Oct, 144(1), 306 - 11
Clostridium perfringens type A: in vitro system for sporulation and enterotoxin synthesis; Smith WP et al.; Polysomes were isolated from an enterotoxigenic strain of Clostridium perfringens during vegetative growth and at 1-h intervals after transfer into Duncan-Strong sporulation medium . During vegetative growth, about 67% of the ribosomes were in polysomal complexes . This proportion decreased to about 20% during the first 2 h in sporulation medium and then gradually increased to a maximum of 45% at 6 h . Ribosomes isolated from cells in vegetative or in sporulation phase could equally translate vegetative, sporulation, and natural viral R17 messenger ribonucleic acid with either vegetative or sporulation initiation factors . When polysomes were allowed to complete their nascent chains with labeled amino acids in vitro, most of the polypeptides synthesized by the vegetative phase and by the sporulation phase polysomes appeared to be identical . There were, however, notable differences upon further investigation . Specifically, when antiserum against the enterotoxin was reacted with the completed polypeptides, no counts were precipitated from the vegetative products . On the other hand, up to 12% of the total labeled protein was precipitated from the products obtained with the sporulation phase polysomes . Upon electrophoresis on sodium dodecyl sulfate, the putative enterotoxin synthesized in vitro ran as a major band with a molecular weight of 35,000, and as two minor bands with molecular weights of 17,000 and 52,000, respectively.

JAMA, 1980 Sep 5, 244(10), 1128 - 30
Clostridium perfringens wound infection associated with elastic bandages; Pearson RD et al.; Clostridium perfringens wound infections were associated with the use of nonsterile elastic outer bandages in diabetic patients who had undergone lower extremity amputation for vascular insufficiency . In each case a second surgical procedure was required . Elastic bandages similar to those used in these procedures were found to contain C perfringens and other clostridial species . This report illustrates the need for maintenance of a sterile, nonpermeable inner barrier to prevent transudation of bacteria into the wound and the potential benefit of using sterile elastic outer bandages after amputation for vascular insufficiency.

Helv Paediatr Acta, 1980 Sep, 35(4), 349 - 57
{Pseudomembranous entercolitis after antibiotic treatment: vancomycin therapy}; Eisenhoffer N et al.; After reporting the case of a child suffering from pseudomembranous colitis (PMC) rapidly cured by vancomycin, the authors present current knowledge which had led to this new mode of therapy . A patient of 2 years 7 months was treated with amoxycillin per os for 8 days for otitis media . A week after this treatment, diffuse abdominal pains appeared, with 8-12 diarrhoeic stools per day containing blood and mucus and recurrent episodes of rectal prolapsus . PMC was diagnosed after rectoscopy and rectal biopsy . The symptomatology persisted for 4 weeks at which time a treatment was begun with vancomycin 4 x 125 mg/day per os . Improvement of the gastrointestinal symptoms occurred 36 hours later, and a new rectal biopsy demonstrated recovery from PMC . Clinical and experimental studies have demonstrated that PMC is due to the proliferation of a toxin producing anaerobic bacterium, clostridium difficile . This organism is highly sensitive to vancomycin . Treatment with vancomycin eliminates clostridium difficile and its toxin from the stools of patients suffering from PMC and permits clinical and histological recovery.

Mikrobiologiia, 1980 Sep-Oct, 49(5), 813 - 20
{Fixation of molecular nitrogen and activity of the microflora in the bottoms of certain lakes in the Estonian SSR and the Rybinsk reservoir}; Saralov AI et al.; The rate of molecular nitrogen fixation was determined in bottom grounds of three Estonian lakes and the Rybinsk water reservoir in the summer of 1977--1978 . Certain species of nitrogen fixing bacteria were found to be confined to lakes of certain trophic type . Ecological niches with the mass growth of Clostridium pasteurianum, Azomonas agilis and Clostridium butyricum were detected in the sediments of eutrophic lakes . Ecological niches of Az . insignis and Cl . acetobutylicum occur in polyhumic lakes . Ecological niches for the nitrogen fixing microaerophilic organisms Methylosinus trichosporium, M . sporium and Mycobacterium flavum can be found in the sediments of water reservoirs of any trophic type . The important ecological factors which favour the growth of microflora with a high rate of molecular nitrogen fixation in bottom grounds are as follows: a sufficiently high temperature, a weakly alkaline reaction, the presence of oxygen in the water near the bottom, and the ratio of organic carbon to total nitrogen about 10--11 . The destruction of organic substances in such sediments involves both aerobic and anaerobic microflora, and Cl . butyricum is abundant among nitrogen fixing microorganisms.

J Infect Dis, 1980 Sep, 142(3), 408 - 13
Epidemiology of experimental enterocecitis due to Clostridium difficile; Larson HE et al.; Hamsters can survive a course of clindamycin if they are held in a protected environment . Inoculation of Clostridium difficile regularly results in fatal enterocecitis in such animals but is without effect in untreated animals . These findings suggest that in the development of enterocecitis, clindamycin treatment and infection with C . difficile are separate events, and they imply that hamsters usually acquire C . difficile from environmental sources . Environments appear to differ in the risk of exposure to C . difficile, high-, medium-, and low-risk areas being recognizable . Once introduced, C . difficile may spread from animal to animal . Parallel with the incidence and epidemiology of human antibiotic-associated pseudomembranous colitis are discussed.

Appl Environ Microbiol, 1980 Sep, 40(3), 571 - 7
Development of ethanol tolerance in Clostridium thermocellum: effect of growth temperature; Herrero AA et al.; The growth of Clostridium thermocellum ATCC 27405 and of C9, an ethanol-resistant mutant of this strain, at different ethanol concentrations and temperatures was characterized . After ethanol addition, cultures continued to grow for 1 to 2 h at rates similar to those observed before ethanol was added and then entered a period of growth arrest, the duration of which was a function of the age of inocula . After this period, cultures grew at an exponential rate that was a function of ethanol concentration . The wild-type strain showed a higher energy of activation for growth than the ethanol-tolerant derivative . The optimum growth temperature of the wild type decreased as the concentration of the ethanol challenge increased, whereas the optimum growth temperature for C9 remained constant . The results are discussed in terms of what is known about the effects of ethanol and temperature on membrane composition and fluidity.

J Clin Microbiol, 1980 Sep, 12(3), 413 - 20
Quantitative aerobic and anaerobic bacteriology of infected diabetic feet; Sapico FL et al.; Quantitative aerobic and anaerobic cultures of deep tissue were performed on amputated infected lower limbs of 13 diabetic patients immediately after surgery . Dissection was made through intact skin distant from any preexisting ulcer . The results were compared with those obtained from: (i) ulcer swabs (pre- and postamputation), (iii) curettage of the ulcer base, and (iii) needle aspiration after normal saline injection . Anaerobic transport media were used for anaerobic cultures before prompt transfer to the anaerobic chamber . A mean of 4.7 bacterial species per specimen was seen (2.3 aerobes, 2.4 anaerobes) . Mean log10 growth per gram of tissue was as follows: (i) aerobes plus anaerobes = 6.99, (ii) aerobes = 6.42, and (iii) anaerobes = 7.65 . There was poor concordance between the deep tissue culture results and the results from other modalities of culture collection, though the results from other modalities of culture collection, though the results from curettage and saline aspiration were slightly better than those obtained from ulcer swabs . The most frequently isolated organisms were enterococci, anaerobic streptococci, and species of Proteus, Clostridium, and Bacteroides . When anti-microbial therapy is indicated for this patient population, the possibility of the concomitant presence of aerobes as well as anaerobes should be considered.

Arch Microbiol, 1980 Sep, 127(2), 167 - 9
Analysis of the fermentation pathways of clostridia using double labelled glutamade; Buckel W; L-(4-14C, 3-3H)Glutamate was used as a tool to elucidate the pathway of its fermentation . The methylaspartate pathway was found in strains of Clostridium tetanomorphum, C . malenominatum, C . limosum, C . lentoputrescens and C . tetani, whereas the hydroxyglutarate pathway was detected in C . sporosphaeroides . These data were supported by measurement of enzyme activities present in cell-free extracts.

Appl Environ Microbiol, 1980 Sep, 40(3), 511 - 5
Heat resistance of the chemical resistance forms of Clostridium botulinum 62A spores over the water activity range 0 to 0.9; Alderton G et al.; Having available the separate chemical resistance forms of Clostridium botulinum 62A spores from an investigation of the effect of spore form on wet heat resistance and also a method for measuring heat resistance at known water activities over the whole water activity (aw) range, we measured the heat resistance of these preparations at four different temperatures at each aw interval of 0.1 from aw 0 to aw 0.9 . The required temperature dependence of resistance was calculated for each aw increment . The spore forms showed a low resistance at aw values of 0 and 0.7 of 0.9, with a rise in resistance in the range aw 0.1 to 0.5 . The temperature dependence values behaved similarly.

Can J Microbiol, 1980 Sep, 26(9), 1162 - 4
A survey of Clostridium perfringens enterotoxin antibody in human and animal sera in western Canada; Niilo L et al.; Sera from human, cattle, sheep, swine, and horse populations in western Canada were tested for the presence of Clostridium perfringens enterotoxin antibody by the passive hemagglutination (PHA) test, supplemented by an immunodiffusion test and by counterimmunoelectrophoresis . A total of 224 human, 345 cattle, 165 sheep, 620 swine, and 768 horse serum samples were examined . Low-titer reactions in the PHA test were detected in human, cattle, horse, and swine sera, in that order, with no titers demonstrated in sheep . The titers in human sera ranged up to 1:128 and three of these samples were also positive in the other two serological tests . The significance of this antibody is not clear, but it is suggested that the low prevalence of the antibody may reflect a low prevalence of enterotoxigenic C . perfringens strains in western Canada . Such serological surveys may be applicable to epidemiological studies involving enterotoxigenic C perfringens.

Can J Microbiol, 1980 Sep, 26(9), 1153 - 7
Ultrastructure of sporulating cells of Clostridium perfringens type A grown in the presence of raffinose; Labbe RG et al.; Seven-hour sporulating cells of Clostridium perfringens type A strain NCTC 10240 grown in the presence of raffinose versus starch were larger, produced a thicker cortex, and were altered in spore coat structure . Strain NCTC 8798 possessed an expanded subcoat region and (or) produced coat fragments when grown in the presence of raffinose.

Infect Immun, 1980 Sep, 29(3), 863 - 72
Interaction of streptolysin O from Streptococcus pyogenes and theta-toxin from Clostridium perfringens with human fibroblasts; Thelestam M et al.; The membrane-damaging properties on human diploid embryonic lung fibroblasts of streptolysin O (from Streptococcus pyogenes) and theta-toxin (from Clostridium perfringens) were compared . The results are consistent with the suggested mechanism for hemolysis by streptolysin O involving one fixation site and one lytic site of this cytolysin . However, the membrane-damaging activity of the two toxins differed with respect to (i) relative cytolytic activity on human diploid lung fibroblasts compared with that on sheep erythrocytes, (ii) binding to the fibroblast membrane, (iii) activity at 0 degrees C, (iv) membrane repair after more than 30 min, and (v) effect on influx of amino acids . It is concluded that the mechanism of membrane damage caused by theta-toxin differs from that of cytoplasmic membrane . These results question the current concept that all thiol-activated, cholesterol-inactivated bacterial toxins are similar both structurally and functionally.

Vopr Med Khim, 1980 Sep-Oct, 26(5), 674 - 7
{Isolation and properties of 3 Clostridium histolyticum collagenases}; Solov'eva NI et al.; The collagenases (I, II and III) have been obtained in a highly purified state from fresh cultural medium of Clostridium histolyticum . The collagenases were similar in their properties to clostridiopeptidase A . The three enzymes differed in their molecular weights, isoelectric points and in some chemical properties . Collagenase II exhibited the most potent hydrolytic activity . Its collagenolytic activity was two-fold higher and the peptidase activity was twenty-fold higher as compared with that of collagenase I . All the three enzymes were inactive towards azocasein and were inhibited by EDTA and cysteine.

Obstet Gynecol, 1980 Sep, 56(3), 311 - 5
A cluster of septic complications associated with illegal induced abortions; Gold J et al.; Between August 7 and October 14, 1977, Clostridium perfringens organisms were isolated from endometrial and/or blood specimens from 3 women who had been hospitalized in McAllen, Texas, after having had illegal induced abortions . One of the women died of clostridial septicemia . A single abortionist was suspected in these 3 cases . The authors investigated the illnesses of these 3 women and those of 6 other women who were hospitalized at some time from January through October 1977 after having had illegal abortions but did not identify a common abortionist . The patient who died was a Medicaid-eligible woman who had had an abortion performed by a nonphysician after public funding of abortion was restricted in Texas as of August 5, 1977 . The authors examined the medical records of all 230 women hospitalized in McAllen General Hospital with abortion complications from 1977 through January 1978 . The local effect of restriction of public funds for abortion was to be studied . The incidence of women admitted to the hospital with febrile abortion complications after August 5, 1977, did not differ from that of women admitted from January 1 through August 4, 1977 . However, the incidence of hospitalization of Medicaid-eligible women with abortion-related complications was greater after August 5 than it was in the earlier period.

Biochim Biophys Acta, 1980 Aug 14, 600(3), 974 - 85
Characterization of membrane permeability alterations induced in Vero cells by Clostridium perfringens enterotoxin; McClane BA et al.; Alterations in plasma membrane permeability induced by Clostridium perfringens enterotoxin were studied using Vero (African green monkey kidney) cells which were radioactively labeled with four markers of different molecular size . The markers were alpha-amino{14C}isobutyric acid (Mr 103), 3H-labeled nucleotide (Mr approx . 300), 51Cr label (Mr approx . 3000) and {3H}RNA (Mr>25000) . Over a 2h period, enterotoxin caused significant release of aminoisobutyric acid, nucleotides and 51Cr label but not RNA . The effects of enterotoxin on label release were dose- and time-dependent . The rate of release of markers was dependent upon their size . Permeability alterations could be detected within 15 min with a high dose of enterotoxin . Gel chromatography of released material was used to determine that markers of Mr 3000 but not 25000 leaked from permeabilized cells . It was concluded that enterotoxin is producing functional 'holes' of limited size in the membrane . Permeability changes due to enterotoxin treatment differed between confluent and nonconfluent (growing) cells . We propose that the primary action of the enterotoxin is to interact with the plasma membrane and produce functional 'holes' of defined size . The resultant alterations in membrane permeability cause the loss of essential cellular substances which inhibits processes such as macromolecular synthesis and eventually leads to cell deterioration and death.

J Biol Chem, 1980 Aug 10, 255(15), 7174 - 80
Purification of carbon monoxide dehydrogenase, a nickel enzyme from Clostridium thermocaceticum; Drake HL et al.; Carbon monoxide dehydrogenase (CO dehydrogenase) has been purified from the homoacetate-fermenting bacterium, Clostridium thermoaceticum . By use of 63Ni, it has been determined that the dehydrogenase is a metallo nickel enzyme . Nickel was rapidly taken up by the organism and most of the ingested metal was found to be incorporated into CO dehydrogenase . As estimated by gel filtration, the native enzyme has a molecular weight of 410,000 . Ferredoxin and a membrane-bound b-type cytochrome, both obtained from C . thermoaceticum, are rapidly reduced by the enzyme in the presence of carbon monoxide and both are considered to be native electron carriers . FMN and Desulfovibrio vulgaris cytochrome c3 were also reduced by the enzyme, while spinach ferredoxin, FAD, NAD, and NADP were not . CO dehydrogenase activity was not appreciably affected by propyl iodide, methyl iodide, carbon tetrachloride, or metal chelators, but was reversibly inhibited by KCN . A method for the in situ assay of CO dehydrogenase in polyacrylamide gels is presented.

Can J Microbiol, 1980 Aug, 26(8), 952 - 8
Stable isotope fractionation by Clostridium pasteurianum . 3 . Effect of SeO32- on the physiology and associated sulfur isotope fractionation during SO32- and SO42- reductions; Harrison GI et al.; Increased SeO32- concentration reduced H2S evolution from SO32- during whole cell and cell-free extract reductions by Clostridium pasteurianum . H2S production from SO42- was completely inhibited by SeO32- in stationary phase cells . Generation times increased with greater SeO32- concentration, the increase with 1 mM SeO32- being a factor of 2.5 for 1 mM SO32-, and over 3 for 1 mM SO42- reductions . In vitro and in vivo experiments with proposed intermediates of the SO32- reduction pathway show that SeO32- inhibited both the S3O62- to S2O32- and S2O32- to S2- reaction sequences with the latter being more pronounced in growth experiments . Both extracts and whole cells reduced SeO32- to Se0 but Se0 granules were not found in the cell's cytoplasm . The formation of S2O32- by an extracellular chemical mechanism appears not to have occurred in these experiments . Increased SeO32- concentration had the effect of compressing the isotopic release pattern for H2S along the H2S production axis and did not significantly alter the maximum and minimum values of delta 34S . Thus, inhibition by SeO32- limited the conversions of sulfur species without altering the isotopic selectivity of rate-controlling steps in the pathway.

J Mol Evol, 1980 Aug, 15(4), 309 - 15
Repeated structure and possible gene duplications in high potential iron protein and rubredoxin; McLachlan AD; The three-dimensional structures of bacterial high potential iron protein (HIPIP) and rubredoxin have been searched for repeats to test whether these molecules evolved by independent tandem gene duplications . HIPIP has no structural repeats in spite of the observed repeated pattern in the amino acid sequence from Rhodopseudomonas gelatinosa . Rubredoxin from Clostridium pasteurianum has repeated hairpin loops of ten alpha-carbon atoms on both sides of the active centre iron-sulphur complex, which can be superposed within a root mean square deviation of 0.84 A by rotating about a local pseudodyad axis . The structural repeat matches a weak repeat in the amino acid sequence . It is concluded that the sequence repeats in HIPIP are probably a coincidence but that rubredoxin may have evolved by gene duplication from a dimer of two primitive hairpin loops.

Pediatrics, 1980 Aug, 66(2), 282 - 4
Aerobic and anaerobic bacteriology of perirectal abscess in children; Brook I et al.; Aspirates of pus from perirectal abscesses in 28 children were studied for aerobic and anaerobic bacteria . A total of 87 isolates (64 anaerobic and 23 aerobic) were recovered from the patients, an average of 2.3 anaerobes and 0.8 aerobes per specimen . Anaerobic organisms alone were recovered from 15 specimens (54%), and in nine specimens (32%) they were mixed with aerobic organisms . Aerobic organisms were recovered in pure culture in only four patients (14%) . The predominant anaerobic organisms were Bacteroides sp (32 isolates, including 14 B fragilis group and seven B melaninogenicus group), Gram-positive anaerobic cocci (15), Fusobacterium sp (six), and Clostridium sp (three) . The predominant aerobic organisms were Escherichia coli and Staphylococcus aureus (six of each), group A beta-hemolytic streptococci, Pseudomonas aeruginosa, and Proteus morganii (two of each) . Five children had a serious underlying chronic disease; three of these had an accompanying bacteremia and two died . Incision and drainage were performed on all patients: some children also received parenteral, or oral antibiotic treatment, or both.

Infect Immun, 1980 Aug, 29(2), 819 - 21
The rat as an animal model for infant botulism; Moberg LJ et al.; Susceptibility to intraintestinal Clostridium botulinum colonization of conventional infant and germfree adult rats is comparable to that of mice . C . botulinum-monoassociated rats pass C . botulinum toxin in their milk.

Infect Immun, 1980 Aug, 29(2), 390 - 4
Evidence that botulinum C2 toxin has two dissimilar components; Iwasaki M et al.; Botulinum C2 toxin produced by most toxigenic and nontoxigenic strains of Clostridium botulinum types C and D contains two distinct protein components, and these were separated by ion-exchange chromatography and gel filtration . Neither of these components manifested the original toxicity, but the original toxicity was restored when the two components were mixed together and trypsinized . This indicates that C2 toxin consists of two dissimilar protein components and that the cooperation of the two is required to elicit toxicity.

Can J Microbiol, 1980 Aug, 26(8), 992 - 7
Electrophoretic analysis of Clostridium botulinum types A and B hemagglutinins; DasGupta BR; The serologically related hemagglutinins of Clostridium botulinum types A and B were resolved into several bands in polyacrylamide gels in the presence of sodium dodecyl sulfate . Reduction of the samples with mercaptoethanol partly changed the band patterns, but with each hemaglutinin the two fastest moving bands in the reduced and unreduced samples were the same . In type A these two smallest units were of molecular weight 14 500 and 19 500; in type B they were of molecular weight 14 500 and 20 000 . The experimentally determined molecular weight of the other resolved units agreed with those calculated on the basis that they were aggregations of the two smallest units in varying but whole multiples.

Am J Vet Res, 1980 Aug, 41(8), 1277 - 9
Clostridium difficile-associated cecitis in guinea pigs exposed to penicillin; Lowe BR et al.; Penicillin treatment resulted in lethal hemorrhagic cecitis in seven of eight guinea pigs . Cecal contents at necropsy from all seven animals contained a cytopathic toxin which was neutralized by Clostridium sordellii and C difficile antitoxins . Bacteriologic cultural examinations of these specimens yielded penicillin-sensitive strains of C difficile which produced a similar or identical cytotoxin in vitro . Stools obtained before penicillin administration and cecal contents from control animals lacked a cytotoxin and cultures failed to yield C difficile . Intracecal injection of cell-free supernatant of C difficile broth cultures reproduced the lesion noted with penicillin treatment . These results implicate C difficile as an agent of penicillin-induced lethal hemorrhagic cecitis in guinea pigs.

Can J Microbiol, 1980 Aug, 26(8), 1018 - 22
Plasmid detection in a bacteriocinogenic strain of Clostridium perfringens; Li AW et al.; Bacteriocinogenic Clostridium perfringens, strain 28, harboured plasmid DNA detectable by dye-bouyant density-gradient centrifugation . This plasmid DNA was absent from an ultraviolet light cured variant which had simultaneously lost its immunity and ability to produce bacteriocin . Agarose gel electrophoresis of the plasmid DNA revealed at least six bands but denaturation experiments suggested three plasmids occurring in more than one conformation . Electron microscopy revealed three major size distributions of circular DNA of molecular weights 1,5,6, and 7.1 megadaltons . Some evidence suggests that the 5.6 megadalton plasmid may control bacteriocin 28 production.

J Med Microbiol, 1980 Aug, 13(3), 445 - 50
Scanning electronmicroscopy of the jejunum in enteritis necroticans; Walker PD et al.; Intestinal tissue resected at laparotomy from patients in Papua New Guinea at various clinical stages of enteritis necroticans, locally known as pig-bel, has been examined under the scanning electronmicroscope . Evidence obtained from parallel studies of experimental infection in pigs is presented . Progressive destruction of the intestinal mucosa was seen during the course of the disease in man . Numerous filamentous rods morphologically consistent with the appearance of Clostridium perfringens type C, were seen to be attached the affected areas of gut and were associated with the necrotic tissue . The mechanism of pathogenicity includes a stage of attachment to the surfaces of jejunal villi, local multiplication, and the production of beta toxin which may be protected from tryptic digestion by the inadequacy of pancreatic protease production in susceptible subjects and by the ingestion of a trypsin inhibitor . The association of the condition with pork feasting is discussed.

Biochim Biophys Acta, 1980 Jul 29, 608(2), 301 - 14
Clostridial apoferredoxin messenger ribonucleic acid . Assay and partial purification; Liao HH et al.; An assay for Clostridium pasteurianum apoferredoxin messenger ribonucleic acid (mRNA) was developed, based on the synthesis of the protein in vitro . Quantitation of apoferredoxin synthesis was accomplished by trypsinization of the cell-free incubation labeled with 3H- or 14C-labeled amino acids, separation of the products by SDS-urea polyacrylamide gel electrophoresis, and excising and counting the NCS-solubilized gel band corresponding to the unique 52-amino acid tryptic peptide derived from apoferredoxin . Its synthesis was shown to be RNA dependent, and was optimized with respect to several parameters of the in vitro protein-synthesizing system . The specificity of the assay was examined with RNA from Clostridium acidi urici, a related species the ferredoxin of which does not yield the 52-amino acid tryptic peptide, and by the use of {3H}leucine, which is not present in C . pasteurianum apoferredoxin . By these methods, the overestimation of apoferredoxin synthesis due to the comigration of fragments from other in vitro products with the legitimate apoferredoxin-derived peptide could be accounted for . The apoferredoxin mRNA was partially purified by the sequential zonal sucrose gradient centrifugation of total RNA followed by Sephadex G-200 chromatography of the enriched RNA, after which a fraction was obtained in which apoferredoxin mRNA was 20-fold enriched . The enriched RNA fraction can now be used for further purification of the apoferredoxin-coding sequences by cloning procedures.

Ann Microbiol (Paris), 1980 Jul-Aug, 131B(1), 11 - 20
{Rate of isolation of "C . difficile" from stools of hospitalized patients: susceptibility of 75 strains (author's transl)}; Tytgat F; Seventy-five strains of Clostridium difficile were recovered from 1,276 stools from patients . Fifty-six of these strains were found to be toxigenic . The rate of isolation of C . difficile was high in culture of stools from patients in surgical intensive care units and in pediatrics units . The susceptibility of these isolates to 13 antimicrobial agents was tested by agar dilution technics . Vancomycin, metronidazole, penicillin and ampicillin at 4 micrograms/ml inhibited all the strains . Cefoxitin, cefotaxim and lincomycin were never active against C . difficile . Only 13.3% of the strains were inhibited by clindamycin at 4 micrograms/ml.

Zh Mikrobiol Epidemiol Immunobiol, 1980 Jul, (7), 43 - 7
{Ultrastructure of Clostridium septicum}; Shakhbanov AA; The study of Cl . septicum by means of electron microscopy revealed that these organisms had peritrichously located flagella and the five-layer cell wall . The formation of small oval spores was observed by the end of the first day . The first sign of sporulation was the appearance of the prespore membrane followed by the formation of the prespore; the formation of the spore coat and the spore cortex began simultaneously at stage IV . The spore coat appeared first as short osmiphil scales . They were located at a certain distance from the outer prespore membrane and had a three-layer structure . Later these scales grew longer and merged, forming an electron-dense layer around the spore . The exosporium had a multilayer structure . Cl . septicum spores had characteristic spiniform processes on the exosporium surface.

Appl Environ Microbiol, 1980 Jul, 40(1), 161 - 2
Stabilities of Clostridium botulinum type B and C toxins in ruminal contents of cattle; Kozaki S et al.; Purified L (large) toxins of Clostridium botulinum type B and C are more stable than M (medium) toxins in ruminal contents of cattle . That finding suggests that the stabilities of L toxins are in part responsible for the incidence of intoxication of cattle.

Am J Dis Child, 1980 Jul, 134(7), 679 - 80
Aerobic and anaerobic bacteriology of pilonidal cyst abscess in children; Brook I et al.; Aspirates of pus from pilonidal abscesses in 25 children were studied for aerobic and anaerobic bacteria . A total of 76 isolates (63 anaerobic and 13 aerobic) were recovered from the patients, accounting for 2.52 anaerobes and 0.52 aerobes per specimen . Anaerobic organisms were recovered from all the specimens, and in eight cases (32%) they were mixed with aerobic organisms . The predominant anaerobic organisms were Bacteroides sp (36 isolates, including ten B fragilis group and ten B melaninogenicus group), Gram-positive anaerobic cocci (16) . Fusobacterium sp (five), and Clostridium sp (four) . The predominant aerobic organisms were Escherichia coli (four) and group D streptococci, alpha-hemolytic streptococci, and Proteus sp (two of each) . We believe this study is the first to demonstrate the mixed anaerobic and aerobic bacteriology of pilonidal cyst abscesses in children.

Avian Dis, 1980 Jul-Sep, 24(3), 658 - 64
Experimental botulism in Pekin ducks; Notermans S et al.; Clostridium botulinum types B, C, and E were found to produce toxin in killed Pekin ducks after oral administration of spores to the live birds . The highest toxin production was found when the ducks were killed within 40 minutes after the administration of the spores . If the ducks were killed after 4 hours, only small amounts of toxin or no toxin at all was found . Pekin ducks were sensitive to botulinum toxin type C-L after oral administration, the LD50 being 9.6 X 10(4) intraperitoneal mouse LD50 . Toxin types B-L, B-M, C-M, and E failed to produce symptoms of botulism . The intravenous LD50 for types B-L, B-M, C-L, and E were respectively 1.5 X 10(4), 1.6 X 10(4), 3.0 X 10(2), and 2.5 X 10(6) intraperitoneal mouse LD50 . Although the results clearly showed that ducks are orally not sensitive for botulinum toxin types B and E, multiplication and toxin production of C . botulinum types B and E in dead ducks is possible . Therefore, such carcasses can be vectors in cycles of these types in the environment.

Postgrad Med J, 1980 Jul, 56(657), 501 - 3
Non-traumatic clostridial myositis: an unusual feature of brain death; Thys JP et al.; In a case of brain death, a Clostridium sordellii and Escherichia coli septicaemia--of probable gastrointestinal origin--developed and was associated with a diffuse clostridial myositis . Factors responsible for the initiation and development of this unusual clostridial infection are discussed.

Antimicrob Agents Chemother, 1980 Jul, 18(1), 13 - 9
Susceptibility of Clostridium botulinum to thirteen antimicrobial agents; Swenson JM et al.; A total of 224 strains of Clostridium botulinum (including isolates from 14 patients with infant botulism and 4 with wound botulism) and 15 strains of C . sporogenes were tested by agar dilution for susceptibility to tetracycline, metronidazole, erythromycin, penicillin, rifampin, chloramphenicol, clindamycin, cephalothin, cefoxitin, vancomycin, sulfamethoxazole-trimethoprim, nalidixic acid, and gentamicin . At least 90% of the C . botulinum strains tested (except for nonproteolytic strains of toxin type F with penicillin) were susceptible to all drugs except sulfamethoxazole-trimethoprim, nalidixic acid, and gentamicin . Minimal inhibitory concentrations for strains from patients with infant and wound botulism were similar to those for other C . botulinum strains.

J Bacteriol, 1980 Jul, 143(1), 274 - 84
In vivo energetics and control of nitrogen fixation: changes in the adenylate energy charge and adenosine 5'-diphosphate/adenosine 5'-triphosphate ratio of cells during growth on dinitrogen versus growth on ammonia; Upchurch RG et al.; The effects of the intracellular energy balance and adenylate pool composition on N2 fixation were examined by determining changes in the energy charge (EC) and the ADP/ATP (D/T) ratio of cells in chemostat and batch cultures of Clostridium pasteurianum, Klebsiella pneumoniae, and Azotobacter vinelandii . When cells of C . pasteurianum, K . pneumoniae, and A . vinelandii in sucrose-limited chemostats were examined, in all cases the EC increased greater than or equal to 15% when the nitrogen source was switched from N2 to NH3 and decreased greater than or equal to 15% when the nitrogen source was switched from NH3 to N2 . The D/T ratio of the same cultures decreased greater than or equal to 70% when they were switched from N2 to NH3 . In such cultures the adenylate pools remained constant when the cells were grown on either NH3 or N2 . In nitrogen (NH3)-limited cultures, the adenylate pool was two- to threefold higher than the adenylate pool in sucrose-limited cultures, and the nitrogenase content of such cells was two- to threefold greater than the nitrogenase content of sucrose-limited N2-fixing cells . The EC and D/T ratio of cells from batch cultures of C . pasteurianum growing on NH3 in the presence of N2 were 0.82 and 0.83, respectively, but when the NH3 was consumed and the cells were switched to a nitrogen-fixing metabolism, the EC and D/T ratio changed to 0.70 and 0.90, respectively . Conversely, when NH3 was added to N2-fixing cultures the EC and D/T ratio changed within 1.5 h the EC and D/T ratio of NH3-grown cells . The nitrogen content of N2-fixing cells to which NH3 was added decreased at a rate greater could be accounted for by cell growth in the absence of further synthesis . This decay of nitrogenase activity (with a half-life about 1.2 to 1.4 h) suggests that some type of inactivation of nitrogenase occurs during repression . The nitrogenase of whole cells was estimated to be operating at about 32% of its theoretical maximum activity during steady-state N2-fixing conditions . Similarities in the data from chemostat and batch cultures of both aerobic and anaerobic N2-fixing organisms suggest that low EC and high D/T ratio are normal manifestations of an N2-fixing physiology.

J Infect Dis . 1980 Jul;142(1):113.
Inhibition of binding of Clostridium difficile toxin by steroids; Chang TW et al.; No detectable inhibition of binding of Clostridium difficile toxin to human erythrocyte lysate was found with alpha-D-(+)-fucose, 1-(-)-fucose, ribose, arabinose, mannose, galactose, xylose, galactosamine, glucosamine, mannosamine, N-acetyl glucosamine, N-acetyl galactosamine, N-acetyl mannosamine, N-acetyl neuraminic acid, lactose, sucrose, lactulose, neuraminidase, saponin, or amantadine . Inhibition was found, however, with a number of sterols and bile acids . In general, bile acids were more active than cholesterol and related compounds . When the bile salts were added greater than or equal to 10 min after exposure to toxin, inhibition was no longer observed . These results suggest that the membrane receptor for C . difficile toxin is not likely to be a carbohydrate component . On the other hand, the inhibitory activity by steroids suggests that a lipid component may play an important role in toxin binding.

J Clin Microbiol, 1980 Jul, 12(1), 18 - 21
Evaluation of a clostridial alpha-toxin disk test for rapid presumptive identification of group B streptococci; Smith JA et al.; An alpha-toxin disk test is described in which group B streptococci completed the hemolysis of sheep erythrocytes partially lysed by the alpha-toxin of Clostridium perfringens . The test was performed satisfactorily on the sheep blood agar primary isolation plate, as well as on pure cultures . A total of 95% of strains of pure group B streptococci tested produced positive reactions within 5 h, and all were positive after overnight incubation, with patterns of synergistic hemolysis readily distinguishable from those seen with group A streptococci . The preparation of disks is well within the scope of most clinical laboratories.

J Gen Microbiol, 1980 Jul, 119(1), 217 - 23
The reduction of nitrate to ammonium by a Clostridium sp . isolated from soil; Caskey WH et al.; Cultures of Clostridium KDHS2 reduced 15NO3- to 15NH4+ with a concurrent increase in molar growth yield of 15.7% compared with fermentatively grown bacteria . The bacteria exhibited a Ks (NO3-) of 0.5 mM and reduced NO3- maximally at a rate of 0.1 mumol h(-1) mg dry wt)-1 . A partially purified nitrate reductase was obtained which had a Km (NO3-) of 0.15 mM . The reduction of 13NO3- to 13NH4+ by resting bacteria was not inhibited by NH4+, glutamate, glutamine, methionine sulphoximine or azaserine . Glutamine synthetase affected neither the synthesis nor the activity of the NO3(-)-reducing enzymes . The results are consistent with the hypothesis that NO3- reduction to NH4+ in this Clostridium sp . is dissimilative . SO32-, but not SO42-, inhibited the reaction, apparently at the level of NO2- reduction.

Med J Aust, 1980 Jun 28, 1(13), 662 - 3
Wound botulism; Fullerton P et al.; It is well recognized that food contamination can result in botulism either from ingestion of performed toxin, in classical botulism, or through absorption of toxin from bacteria within the gut, in infant botulism . Botulism due to contamination of wounds with Clostridium botulinum is not commonly recognized . We report a case of wound botulism occurring in an eight-year-old boy, characterized by early ptosis, dysphagia and dysarthria and then followed by progressive generalized paralysis and fixed dilated pupils, but with intact sensorium . Management consisted of early wound debridement and prolonged intensive respiratory and nutritional support . Recovery was complete.

J Biol Chem, 1980 Jun 10, 255(11), 5004 - 6
Chemical characterization of an alkali-labile bond in the polypeptide of proline reductase from Clostridium sticklandii; Seto B; In previous studies, it was shown that a 4600-dalton pyruvate-containing peptide is released from proline reductase by mild alkali treatment . The alkali-sensitive bond proved to be between Ser-Glu, and it was suggested that an ester, rather than a peptide linkage, might be involved . In the present study, the effects of additional esterolytic reagents, (I) LiBH4 and (II) NH2OH, on proline reductase have been investigated and compared with 0.1 N NaOH-induced cleavage . Treatment with reagents I and II released a peptide identical with the peptide released by alkali as judged by electrophoretic mobility on thin layer sheets, COOH-terminal analyses, and amino acid compositional studies . The glutamic acid residue is converted to alpha-amino-delta-hydroxyvaleric acid after reductive cleavage with LiBH4 . The liberation of the peptide fragment by the relatively specific esterolytic reagent, LiBH4, provides additional support of the presence of an ester linkage in native proline reductase.

J Biol Chem, 1980 Jun 10, 255(11), 5364 - 72
Purification to homogeneity of H blood group beta-galactoside alpha 1 leads to 2 fucosyltransferase from porcine submaxillary gland; Beyer TA et al.; A beta-galactoside alpha 1 leads to 2 fucosyltransferase has been solubilized from porcine submaxillary glands and purified 124,000-fold to homogeneity by repeated affinity chromatography on GDP-hexanolamine agarose . Sodium dodecyl sulfate polyacrylamide gel electrophoresis of the purified enzyme revealed two electrophoretic species with apparent Mr = 60,000 and 55,000 . The two enzyme species have not been completely resolved, but both appear to be active forms of the fucosyltransferase with approximately equal specific activities . Glycosidase digestion of the fucosylated products with the alpha 1 leads to 2-specific fucosidase from Clostridium perfringens and the alpha 1 leads to 3/alpha 1 leads to 4-specific fucosidase from almond emulsin indicates that the enzyme forms exclusively the Fuc alpha 1 leads to 2Gal linkage with a variety of acceptor substrates . A GDP-fucose hydrolase activity co-purifies with the fucosyltransferase . Identical rates of thermal inactivation and co-migration on gel electrophoresis under nondenaturing conditions suggest that the two activities are due to a single enzyme species.

Zentralbl Bakteriol A, 1980 Jun, 247(1), 95 - 100
{Phage conversion of Clostridium novyi type A (author's transl)}; Schallehn G et al.; Clostridium novyi type A, the cause of gas gangrene in man and animal, produces four main toxic components (alpha, gamma, delta, and epsilon) . The alpha toxin is a necrotizing toxin produced in lethal amounts by strains of type A . This lethal toxin induces a characteristic subcutaneous colorless gelatineous edema when guinea pigs are infected with C . novyi type A . In this paper we report further evidence on the relationship of bacteriophages to the production of alpha toxin by C . novyi type A . The results showed that five phage-sensitive bacterial strains that did not produce alpha toxin could be isolated from alpha toxin producing stains . These nontoxigenic strains could be infected with specific bacteriophages isolated from the different toxigenic strains of C . novyi type A and converted to alpha toxin production . The toxigenicity of these converted strains depended upon the continued participation of specific bacteriophages . These bacteriophages were shown to be very unstable in culture supernatant fluids.

Aust Vet J, 1980 Jun, 56(6), 285 - 91
Levamisole vaccine combinations . 1 . Heightened antibody response; Hogarth-Scott RS et al.; Levamisole was combined with a clostridial vaccine and experiments conducted in sheep to determine the effect on anthelminitic and vaccine efficacy . Vaccine potency was assessed in separate experiments in 200 sheep (divided into 5 groups) . The individuals in each group were injected with one of the following 5 different preparations: levamisole combined with anaerobic vaccine at pH of either 3.5 or 6.0, vaccine alone at a pH of either 3.5 of 6.0, and levamisole alone . The combined levamisole/vaccine preparations had been stored for not less than 6 months prior to testing . A statistically-significant heightened antibody response to 5 antigenic components in the vaccine was obtained when the pH of the vaccine was reduced to 3.5 and the vaccine was given in combination with levamisole . This trend was present in the antibody response of the sheep when a pooled sample of the individual serums was tested at 6 weeks, 8 weeks and 14 weeks (8 weeks after the second injection) . From the analysis of the individual serums at week 8 (2 weeks after the 2nd injection) there was evidence that the distribution curve of the titres had shifted to the right suggesting that the zero or poor responders in the population had improved immunocompetence, with the better responders also improving proportionately.The immunogenicity of the blackleg component was tested in sheep, 9 weeks and 32 weeks after the second injection and gave complete protection to a lethal challenge of Clostridium chauvoei . This combination offers not only increased ease of administration by providing a single anthelmintic and anaerobic vaccines injectable preparation (probably treatment with either or both is the two most common and important veterinary procedures undertaken by farmers, but in addition, the antibody titre of the treated animals is significantly increased . Part 2 of this paper establishes that the anthelmintic efficacy of the levamisole/vaccine combination was not impaired and that tissue reactions at the site of injection were acceptable.

Infect Immun, 1980 Jun, 28(3), 1041 - 3
Production of Clostridium difficile antitoxin; Ehrich M et al.; We have produced antitoxin to the toxin of Clostridium difficile in rabbits and in goats . Antitoxin dilutions of 1/8,000 and 1/5,120 were capable of neutralizing lethal doses of the toxin in mice and in tissue culture, respectively.

J Infect Dis, 1980 Jun, 141(6), 772 - 4
Colitis associated with metronidazole therapy; Saginur R et al.; A 46-year-old woman was treated with oral metronidazole for trichomonal vaginitis and developed diarrhea, which persisted for five weeks . Tissue culture assay of stool supernatant showed a cytopathic toxin that was neutralized by Clostridium sordellii antitoxin, and cultures yielded Clostridium difficile, which produced a similar or identical cytotoxin in vitro . This isolate proved sensitive to metronidazole at 0.25 microgram/ml . Prior reports have indicated that metronidazole may also be used therapeutically in patients with antibiotic-associated colitis ascribed to other agents . The patient presented here shows the enigma that the same agent used for therapy of colitis may also cause this complication.

Arch Intern Med, 1980 Jun, 140(6), 834 - 5
Neutropenic enterocolitis and Clostridium septicum infection in patients with agranulocytosis; Rifkin GD; Two patients with previously undiagnosed agranulocytosis had rapidly fatal Clostridium septicum sepsis and neutropenic enterocolitis . This toxigenic organism has a known predilection for cecal lesions . The association reported herein suggests that C septicum may be the cause of this necrotizing enteropathy.

Am J Dis Child, 1980 Jun, 134(6), 564 - 6
Chronic otitis media in children . Microbiological studies; Brook I; Aspiration of the exudate through the open perforation was performed in 68 children with chronic otitis media . The middle ear aspirate and a swab specimen of the external auditory canal were cultured aerobically and anaerobically . Seventy-eight isolates were recovered from the middle ears, 99 from the external ear canals, and 95 were present at both sites . Aerobes only were isolated from 33 patients (48.5%), nine (13.2%) had only anaerobes, and 26 (38%) had a culture that grew both aerobes and anaerobes . There were 99 aerobic isolates . Aerobes commonly recovered were Pseudomonas aeruginosa, Staphylococcus aureus, Proteus sp, Klebsiella pneumoniae, and Haemophilus influenzae . There were 74 anaerobes isolated . Anaerobes commonly isolated were anerobic Gram-positive cocci, Bacteroides sp, and Clostridium sp . These findings demonstrate the polymicrobial bacteriology of chronic otitis media in children . Cultures collected from the external auditory canals prior to their sterilization can be misleading . Reliable information can be obtained from the ear exudates when collected through the open perforation in the tympanic membrane.

Gastroenterology, 1980 Jun, 78(6), 1584 - 6
Bacitracin treatment of antibiotic-associated colitis and diarrhea caused by Clostridium difficile toxin; Chang TW et al.; Four cases of antibiotic-associated colitis and diarrhea caused by Clostridium difficile were successfully treated with oral bacitracin, 25,000 units four times daily for 7-10 days . Diarrhea resolved in all of the cases, in 2 days, with disappearance of Clostridium difficile toxin in the stools in 3 out of 4 patients so measured . Two of the patients treated had relapses after vancomycin, while the other 2 were experiencing the first episodes . One patient relapsed after bacitracin treatment, but was treated successfully with vancomycin . Our preliminary experience indicates that bacitracin, being less expensive and more readily available world-wide than vancomycin, could be used as an alternative drug for toxin-induced colitis or diarrhea.

J Clin Microbiol, 1980 Jun, 11(6), 604 - 9
Cultural and physiological characteristics and antimicrobial susceptibility of Clostridium botulinum isolates from foodborne and infant botulism cases; Dezfulian M et al.; Isolates Clostridium botulinum from foodborne and infant botulism cases in the United States were compared on the basis of toxigenicity, cultural and biochemical characteristics, metabolic products, and susceptibility to antimicrobial agents . Seventy-eight strains, including 42 from foodborne and 36 from infant botulism sources, were examined . Cultures on anaerobic blood agar exhibited circular, spindle, and rhizoid (medusa head) colonies . Overall, the characteristics of isolates from foodborne and infant botulism cases were quite similar . We concluded that it was not possible to differentiate C . botulinum isolates associated with foodborne botulism from those recovered from infant botulism cases . All of the 78 strains produced an unidentified indole derivative(s), detected with paradimethylaminocinnamaldehyde reagent, and hydrocinnamic acid, detected by gas-liquid chromatography; all exhibited a high degree of resistance to cycloserine, sulfamethoxazole, and trimethoprim . These characteristics should prove to be useful in the isolation and identification of C . botulinum from mixed microbial populations.

Gut, 1980 Jun, 21(6), 493 - 9
Mucosal damage mediated by clostridial toxin in experimental clindamycin-associated colitis; Abrams GD et al.; A toxin produced by Clostridium difficile has been implicated in the pathogenesis of antibiotic-associated colitis in humans and experimental animals . This study was undertaken in order to define the sequential evolution of caecal mucosal lesions in the hamster and to relate those lesions directly to the clostridial toxin . Sterile filtrates from a culture of C . difficile and from caecal contents of clindamycin-treated hamsters were studied with respect to their effects on the caecal mucosa and on cultured cell monolayers . The toxic filtrates both produced cellular swelling in vitro, and appeared to have a similar cytotoxic effect on caecal epithelial cells in vivo . Cellular damage was followed by extensive epithelial desquamation and the evolution of an acute pseudomembranous typhlitis . The pathogenetic sequence produced by the filtrates was identical with that previously described after direct clindamycin treatment . These findings demonstrate that intraluminal clostridial toxin can mediate development of the characteristic antibiotic-associated mucosal lesions.

J Immunol, 1980 Jun, 124(6), 2615 - 9
Genetic control of the immune response to ferredoxin: linkage and mapping of T cell proliferation and antibody production genes to the MHC of mice; Sikora LK et al.; The genetics of the immune response in the mouse were studied by using the antigenically simple, stable, naturally occurring protein ferredoxin (Fd) from Clostridium pasteurianum . The immune status of mice primed and boosted with Fd was assessed by using two parameters of immunity: T cell proliferation and serum antibody production with the ELISA method . In both assay systems, the response has been shown to be H-2 linked: k, b, and s haplotypes respond to Fd, and H-2d mice are nonresponders . It is apparent that different immunoregulatory events modulate the response in the responder strains; these factors become evident in the recombinant analysis of the response and to date an immunoregulatory gene(s) has been mapped to at least the K/I-A subregions . F1 analysis demonstrated a gene dose-dependent response of the strains studied.

Antimicrob Agents Chemother, 1980 Jun, 17(6), 957 - 60
Susceptibilities of anaerobic bacteria to cefoperazone and other antibiotics; Kaye D et al.; Two hundred fifty clinical isolates of anaerobic bacteria were tested for suceptibility to cefoperazone, cefamandole, cefoxitin, carbenicillin, clindamycin, and chloramphenicol . Anaerobic gram-positive cocci were susceptible to all of the antibiotics tested . Clindamycin was the most active agent against Bacteroides species, followed by chloramphenicol and then cefoxitin . Cefoperazone was less active than cefoxitin and equal in activity to carbenicillin . Cefamandole was the least active antibiotic against Bacteroides . B . distasonis, B . vulgatus, B . thetaiotaomicron, and B . ovatus were more resistant to the antibiotics than B . melaninogenicus, B . oralis, or B . bivius . Clindamycin was the most active agent against Clostridium species, followed by chloramphenicol; the three cephalosporins and carbenicillin were about equal in activity . Clindamycin was the most active antibiotic against Fusobacterium species, followed by chloramphenicol, carbenicillin, and cefoperazone (which were about equally active) and then cefamandole.

Antimicrob Agents Chemother, 1980 Jun, 17(6), 1004 - 7
Therapy of obstetrical infections with moxalactam; Gibbs RS et al.; We evaluated moxalactam in 62 patients with puerperal or postabortal genital infections . In all patients, the initial dose was 6 g/day . In 84% of patients, we found anaerobes in genital specimens . Of aerobic isolates, only enterococci were resistant . Among anaerobes tested, only two isolates (a Clostridium leptum and a Bacteroides disiens) had minimal inhibitory concentrations of greater than or equal to microgram/ml . Good clinical responses occurred in 56 of 62 (90%) . Moxalactam was well tolerated with little local irritation and minimal hepatic, renal, or hematological abnormalities.

J Hyg (Lond), 1980 Jun, 84(3), 443 - 56
Application of serological typing to the investigation of outbreaks of Clostridium perfringens food poisoning, 1970-1978; Stringer MF et al.; Serological typing was used as an epidemiological tool in the investigation of 524 outbreaks of Clostridium perfringens food poisoning in the United Kingdom and 37 outbreaks in other countries . Five thousand five hundred and fifty-four (77%) of 7245 strains of C . perfringens associated with the 561 outbreaks were typable with the 75 Food Hygiene Laboratory antisera; in 354 (63%) of these outbreaks a specific serotype was established as being responsible for the outbreak . An assessment is made of the ability of two additional sets of antisera, prepared against 34 American and 34 Japanese strains of C . perfringens, to increase the number of strains which can be typed . The extent of cross-reaction between the three sets of antisera was determined and the results are discussed in relation to the source and history of the type strains.

Br J Exp Pathol, 1980 Jun, 61(3), 261 - 71
Experimental pigbel: the production and pathology of necrotizing enteritis due to Clostridium welchii type C in the guinea-pig; Lawrence G et al.; An animal model for pigbel in man was developed using guinea-pigs . Intragastric dosing with growing cultures of Clostridium welchii Type C only produced necrotic lesions if protease inhibitors were given as well . beta toxin, which is made by the Type C organism, causes the intestinal damage and is very easily destroyed by proteases . Protease inhibitors in soybean and aprotinin were effectively in inducing disease in animals on a normal diet, while inhibitors in sweet potato, which inhibit only trypsin, were only effective in animals on a low-protein diet . In experiments using intragastric dosing, and in those where cultures and toxic filtrates were injected directly into the jejunum, the animals could be protected with an excess of pancreatic enzymes or by active or passive immunization against beta toxin . The pathology off Type C necrotizing enteritis in guinea-pigs had the macroscopic and microscopic features of pigbel in man . These experiments suggested the basic importance of a low-protein diet and dietary trypsin inhibitors in the pathogenesis of pigbel in man.

Appl Environ Microbiol, 1980 Jun, 39(6), 1120 - 2
Improved method for purification of enterotoxin from Clostridium perfringens type A; Granum PE et al.; The purification procedure of Clostridium perfringens type A enterotoxin has been improved . The cell sonic extract was precipitated twice with ammonium sulfate, first 40% saturated to concentrate the enterotoxin and then 15% saturated . The two precipitations were followed by gel filtration on Sephadex G-100 . The enterotoxin appeared to be homogeneous on 7% polyacrylamide gel electrophoresis after this three-step purification procedure, with a recovery of 56% and a 12.3-fold purification . The solubility properties at different pH values, temperatures, and ammonium sulfate concentrations are also given as basis for the purification procedure.

Zentralbl Bakteriol A, 1980 Jun, 247(1), 101 - 13
{Characterization of bacteriophages of Clostridium novyi type A (author's transl)}; Imhoff D et al.; Four bacteriophages of Clostridum novyi type A (PFo, P5771, PA1350e and P19402) were examined . The phages were spontaneously released to the culture medium in titers of 10(6) to 10(8) pfu/ml at the end of the bacterial growth of the donor strain . The phage titer could be increased to 10(9) to 10(12) pfu/ml by growing the phages in the culture of the indicator strain C . novyi 5771/HS 10 . These high titered phage suspensions were used for morphological studies and for the production of anti-phage-sera . The phages of C . novyi were unstable and lost most of their infectivity within 24 h . Lyophilizing the phages in glutamate medium seemed to be one possible way of partially stabilizing these phages . Phages PFo, P5771, PA1350e and P19402 were similar in morphology and size, in antigenic pattern and in plaque morphology . Phage PA1350e was stabile only at pH 7 and below 40 degrees C for a short time . It was inactivated at 50 degrees C within 20 min, at 55 degrees C and at 60 degrees C in 4 min.

Biochim Biophys Acta, 1980 May 29, 623(1), 84 - 8
Photo-oxidation of L-glutamate decarboxylase from Escherichia coli, sensitized by the coenzyme pyridoxal phosphate and by proflavin; Cozzani I et al.; Irradiation of L-glutamate decarboxylase (L-glutamate 1-carboxy-lyase, EC 4.1.1.15) from Escherichia coli by visible light absorbed by the intrinsic chromophore, pyridoxal phosphate, caused the selective modification of two methionines per enzyme monomer . The disulfoxide derivative exhibited modified circular dichroism, chromatographic and kinetic properties, suggesting a conformational role for the two methionine residues . Irradiation of the enzyme in the presence of proflavin revealed the presence of two distinct groups of tryptophan residues with markedly different photooxidation rate constants . No evidence of involvement of tryptophans in the catalytic mechanisms of the enzyme was obtained . The results are compared with those obtained on irradiation of L-glutamate decarboxylase from Clostridium perfringens.

Biochim Biophys Acta, 1980 May 29, 623(1), 124 - 38
Nitrogenase XII . Mössbauer studies of the MoFe protein from Clostridium pasteurianum W5; Huynh BH et al.; We have studied the molybdenum-protein (MoFe protein) from Clostridium pasteurianum with Mossbauer spectroscopy in the temperature range from 1.5 to 200 K in magnetic fields up to 55 kG . Except for some small differences in the hyperfine parameters the results for the C . pasteurianum protein are essentially the same as those published previously for the protein from Azotobacter vinelandii, i.e . (30 +/- 2) Fe atoms partition into two identical cofactor centers M (each center most likely containing six Fe atoms and one Mo atom), four P-clusters (each center containing four Fe atoms), and one iron environment labeled S (about two Fe atoms per holoenzyme) . We have analyzed the spectra of the cofactor centers in three distinct oxidation states, Formula: (see test) . The diamagnetic (electronic spin S = 0) state MOX is attained by oxidation of the native, EPR-active (S = 3/2) state MN . The reduced state MR is observed in steady state under nitrogen fixing conditions; high-field Mossbauer studies show that the cofactor centers are paramagnetic (integer electronic spin S greater than or equal to 1) in the state MR . We have evaluated the complex high-field spectra resulting from the P-clusters in the oxidized state POX . The analysis shows that one iron site is characterized by a positive hyperfine coupling constant A0 while the other three sites have A0 less than 0 . A slightly modified set of parameters also fits the high-field data of the MoFe protein from A . vinelandii . Finally, we will present a discussion summarizing our principle results obtained to date for the proteins from A . vinelandii and C . pasteurianum.

Infect Immun, 1980 May, 28(2), 303 - 9
Oral toxicities of Clostridium botulinum type C and D toxins of different molecular sizes; Ohishi I et al.; Clostridium botulinum type C progenitor toxins of different molecule sizes, C-L (16S) and C-M (12S), were purified from cultures of strains 573, Stockholm, and CB-19 . C-L toxin showed some hemaggglutinin activity, whereas C-M toxin did not . Neither C-L nor C-M toxin was activated upon trypsinization . Molecular dissociation of purified type C-L and C-M toxins into toxic and nontoxic components was demonstrated by sucrose density gradient ultracentrifugation and diethylaminoethyl-Sephadex chromatography at pH 8.0 . The molecular construction of type C progenitor toxin appears to be analogous to that reported for botulinum toxins of other types . C-L and D-L toxins showed higher oral toxicities to mice than did C-M or D-M toxin . Such higher oral toxicities were ascribed to the higher stabilities of these toxins in gastric and intestinal juices.

Eur J Biochem, 1980 May, 106(2), 439 - 47
The reversible dehydration of (R)-2-hydroxyglutarate to (E)-glutaconate; Buckel W; 1 . During fermentation with whole cells of Acidaminococcus fermentans or Clostridium microsporum the pro-3S hydrogen of (R)-2-hydroxyglutarate or of its precursor (S)-glutamate is eliminated stereospecifically . Since (E)-glutaconate but not its Z isomer is fermented by whole cells or cell-free extracts of A . fermentans, the overall dehydration of (R)-2-hydroxyglutarate to (E)-glutaconate can be described as syn . 2 . The fermentation of (E)-glutaconate required acetyl phophate, CoA and NAD, that of (S)-glutamate or (R)-2-hydroxyglutarate additionally MgCl2, FeSO4 and dithioerythritol . The fermentations of all three substrates were inhibited by avidin and stimulated by biotin . 3 . The hydration of (E)-glutaconate was measured enzymically by the formation of (R)-2-hydroxyglutarate . The dehydration of the hydroxy acid was assayed by the release of 3HOH from (2R)-2-hydroxy{3-3H}glutarate . Optimum conditions were found by activation of the cell-free extract with MgCl2, FeSO4, dithioerythritol, acetyl phosphate anmd NADH followed by the reaction which only required acetyl phosphate and CoA as cofactors . Activation and reaction had to be performed anaerobically . 4 . The dehydration was inhibited by 2 mM azide, 1 mM arsenate, 1 mM hydroxylamine, 20 micro M dinitrophenol or 10 micro M carbonylcyanide p-trifluoromethoxyphenylhydrazone . 5 . It is concluded that the actual substrates of the dehydration are the corresponding thiol esters . The data indicate a catalytical phosphorylation during the reaction.

Appl Environ Microbiol, 1980 May, 39(5), 943 - 9
Effect of moderately acidic pH on heat resistance of Clostridium sporogenes spores in phosphate buffer and in buffered pea puree; Cameron MS et al.; The effect of pH in the range 5.0 to 7.0 on the thermal destruction of spores of Clostridium sporogenes putrefactive anaerobe 3679 was examined by three methods: a capillary tube method in which spores were suspended in phosphate buffers, a thermoresistometer method in which spores were suspended in buffered pea puree adjusted to the same set of pH values, and a thermal death time can method in which spores were again suspended in buffered pea puree . The results indicated that increasing acidity is, in general, accompanied by decreasing heat resistance, although the pH effect was more pronounced at the higher than at the lower processing temperatures . Certain pH values appear to be critical, as they produced, in all three sets of experiments, effects which would not be predicted by the overall relationship between acidity and spore heat resistance . Differences between heat resistance in phosphate buffer as compared with that in pea puree adjusted to the same pH were also noted . D-values in buffer were found to be lower than those in pea puree, except at the highest temperatures coupled with the lowest pH values . The differences between buffer D-value and pea puree D-value were found to increase with increasing pH and with decreasing temperature . On the other hand, at all pH values examined, z-values determined in buffer were somewhat higher than those determined in pea puree adjusted to the same pH.

Acta Pathol Jpn, 1980 May, 30(3), 445 - 50
A report on the pathology of type A botulism; Toyoda H et al.; An autopsy case of type A botulism, a sacrifice during the first outbreak in Japan, was presented . The patient exhibited typical neuromuscular symptoms and died on the 8th day from the onset of illness . Type A Clostridium botulinum was demonstrated from the stools and the toxin from the blood serum during the course of disease . The infection source of botulinus bacilli, however, was not clarified . The autopsy diagnoses were made as follows; 1) type A botulism, 2) bronchopneumonia, 3) respirator brain, 4) congestion, 5) hemorrhage in vagal nerve, myocardium and endometrium, 6) gastric erosion, 7) cloudy swelling of kidneys, 8) enterocolitis, 9) focal necrosis of liver and adrenals, 10) demyelination of cranial nerves, and 11) focal hyaline degeneration of striated and smooth muscles and myocardium . The botulinus toxin was confirmed from the samples of cardiac blood or autopsy.

J Clin Microbiol, 1980 May, 11(5), 470 - 3
Identification of toxigenic Clostridium difficile by counterimmunoelectrophoresis; Welch DF et al.; A counterimmunoelectrophoresis (CIE) technique which reacted positively with culture filtrates of Clostridium difficile was developed and compared with a cytotoxicity assay in human embryonic lung cell cultures . CIE, employing C . sordellii antitoxin, detected 17 of 17 C . difficile strains . Of those positive by CIE, 13 were cytotoxic in cell culture . Fourteen Clostridium species other than C . difficile, C . sordellii, and C . bifermentans were negative by CIE . C . sordellii and C . bifermentans gave positive CIE results but were not cytotoxic . Similar sensitivity of toxin detection was observed for both methods . Optimal conditions for performing CIE included use of 48-h chopped meat-glucose broth cultures as the antigen source, use of a 10x-concentrated U.S . Standard C . sordillii antitoxin, and electrophoresis for 1.5 h in 0.05 M tris(hydroxymethyl)aminomethane-barbital-sodium barbital, pH 8.8, at a constant current of 6 mA/slide . CIE appears to be a suitable alternative to the cytotoxicity assay and may serve as a means for presumptive identification of C . difficile.

Mikrobiologiia, 1980 May-Jun, 49(3), 427 - 32
{Regulation of nitrogenase activity and biosynthesis in Clostridium butyricum}; Saralov AI; The effect of various factors on the activity and biosynthesis of nitrogenase in Clostridium butyricum was estimated by the rate of acetylene reduction at different growth phases in the static culture . The activity of nitrogenase was found to be low in vegetative cells; it increased and reached the maximum at the stage of prospore formation and decreased in the course of sporulation . The duration of different stages in the cell growth cycle depended on temperature and the composition of the medium . An increase in the rate of nitrogenase synthesis at the stage of prospore formation was favoured by the presence of sodium acetate or yeast autolysate in the medium; ammonium chloride inhibited the mechanism of nitrogenase biosynthesis . Cytoplasmic membranous structures such as tubular-vesicular or lamellar mesosomes were formed in active nitrogen fixing cells grown in a medium with yeast autolysat . Such structures were absent from cells with a low activity grown in a medium containing ammonium chloride.

Hoppe Seylers Z Physiol Chem, 1980 May, 361(5), 697 - 702
{1H-NMR-spectroscopic evidence for the release of N-acetyl-alpha-D-neuraminic acid as the first product of neuraminidase action (author's transl)}; Friebolin H et al.; The 1H-NMR spectroscopy was used to study the anomeric configuration of N-acetyl-D-neuraminic acid released by the action of neuraminidase . The hydrolysis of NeuAcalpha 2 leads to 3 Gal-beta 1 leads to 4Glc (20mM) by the enzymes of Clostridium perfringens and Arthrobacter ureafaciens (50 mU, 150 mU and 800 mU, respectively) in 50mM Na/K-phosphate buffer pD 5.4 was observed by recording the spectra . On the basis of the characteristic signals of the protons at C-3 (alphaNeuAc: delta{H(3e)} = 2.72, delta{H(3a)} = 1.64; betaNeuAc: delta{H(3e)} = 2.25, delta{H(3a)} = 1.84) the product of the enzymatic cleavage was identified to be the N-acetylneuraminic acid in the alpha-anomeric form . Two hypotheses are discussed to explain how the enzymatic hydrolysis may occur and how N-acetyl-alpha-D-neuraminic acid leaves the catalytic site of the neuraminidases with retention of the C-2 configuration.

Antimicrob Agents Chemother, 1980 May, 17(5), 901 - 4
Comparison of moxalactam (LY127935) and cefotaxime against anaerobic bacteria; Jorgensen JH et al.; The in vitro activities of moxalactam (LY127935 {6059S}) and cefotaxime were compared with those of cefoxitin, cefamandole, cefuroxime, carbenicillin, and penicillin by agar dilution susceptibility testing of a variety of anaerobic bacteria . Moxalactam proved to be the most active agent tested against Bacteroides fragilis and other species of the B . fragilis group . Moxalactam and cefotaxime showed activity similar to the other drugs against the remaining species of Bacteroides, Fusobacterium, Actinomyces, Propionibacterium, and Veillonella . Penicillin was the most effective drug tested against most species of Clostridium, the anaerobic gram-positive cocci, and Eubacterium lentum.

MMW Munch Med Wochenschr, 1980 Apr 25, 122(17), 633 - 6
{Metronidazole sensitivity of anaerobes . A comparison with other chemotherapeutics (author's transl)}; Werner H et al.; In comparative in vitro studies, broad spectrum penicillins and cephalosporins showed a relatively weak action in the range of beta-lactamase-positive bacteroides species (B . fragilis, B . thetaiotamicron) bacteroidaceae activity being otherwise good . Clindamycin was more effective against bacteroides species than against bacteria of the fusobacterium-sphaerophorus group . Metronidazole, however, showed no gap in its efficacy against bacteroidaceae, peptococcaceae and clostridium spp . with the exception of Propionibacterium acnes . The substance is therefore suitable alone and in combination for the chemotherapy of anaerobic infections.20

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