Med Microbiol Immunol (Berl), 1984, 173(1), 49 - 55 Interaction between Clostridium difficile and polymorphonuclear leucocytes from the elderly and post-operative cancer patients: phagocytosis and bactericidal function; Bassaris HP et al.; The ability of polymorphonuclear leucocytes (PMNs) from elderly healthy subjects and post-operative cancer patients to ingest and kill Clostridium difficile was studied . The percentage of phagocytosis in clinically healthy subjects aged 69-82 years ranged from 20.2 +/- 3.2 (mean +/- SD) to 34.1 +/- 4.2, depending on the strain of C . difficile, from 6.7 +/- 2.2 to 11.2 +/- 2.2 in post-operative cancer patients aged 65-69, and from 68.4 +/- 3.1 to 81.1 +/- 6.3 in young healthy control subjects . In both study groups, the defect was in part serum-associated . Although the total number of bacteria killed by the PMNs was reduced in the two study groups, the percentage of ingested bacteria killed by the PMNs was similar to that in the young healthy subjects . These differences were not noted when Escherichia coli and Staphylococcus aureus were used as the test organisms . The defective PMN function against C . difficile in the elderly and post-operative cancer patients described in this study may offer an explanation as to why these individuals are at high risk of developing antibiotic-associated colitis. J Mol Evol, 1984-85, 21(4), 339 - 45 Examination of protein sequence homologies: I . Eleven Escherichia coli L7/L12-type ribosomal "A" protein sequences from eubacteria and chloroplast; Otaka E et al.; Seven complete and four partial sequences of Escherichia coli L7/L12-type ribosomal "A" proteins obtained from various bacteria (E . coli, Bacillus subtilis, Micrococcus lysodeikticus, Rhodopseudomonas spheroides, Desulfovibrio vulgaris, Streptomyces griseus, Bacillus stearothermophilus, Clostridium pasteurianum, Arthrobacter glacialis, and Vibrio costicola) and spinach chloroplast have been reexamined using a computer program that searches for homologous tertiary structures . Comparison matrices for the sequences show that they match the sequence of E . coli L7 (EL7) if one assumes the insertion or deletion of certain residues at sites corresponding to residues 1, 38, 49, and 92 of EL7 . That two additional insertion points are found only in the spinach chloroplast protein suggests that the chloroplast protein probably diverged from the bacterial forms . Further phylogenetic relationships among these 11 prokaryote-type "A" proteins are discussed with respect to average correlation coefficients computed, taking into account the existence of the gaps. Can J Comp Med, 1984 Jan, 48(1), 111 - 2 An enzyme-linked immunosorbent assay for the detection of Clostridium perfringens enterotoxin antibody; Niilo L et al.; An enzyme-linked immunosorbent assay (ELISA) was adapted to test serum antibody to enterotoxin of Clostridium perfringens type A . The test was evaluated using sheep, calf and guinea pig sera and compared with passive hemagglutination and immunodiffusion tests . The ELISA was found to be more sensitive than the other two tests and was completely free from nonspecific reactions . The method was considered to be technically advantageous and suitable for semiautomated procedures. Arch Microbiol, 1984 Jan, 137(1), 63 - 9 The synthesis of acetyl-CoA by Clostridium thermoaceticum from carbon dioxide, hydrogen, coenzyme A and methyltetrahydrofolate; Pezacka E et al.; It has been demonstrated that enzymes from Clostridium thermoaceticum catalyze the following reaction in which Fd is ferredoxin and CH3THF is methyltetrahydrofolate . (for formula see text) . The system involves hydrogenase, CO dehydrogenase, a methyltransferase, a corrinoid enzyme and other unknown components . Hydrogenase catalyzes the reduction of ferredoxin by H2; CO dehydrogenase then uses the reduced ferredoxin to reduce CO2 to a one-carbon intermediate that combines with CoASH and with a methyl group originating from CH3THF to form acetyl-CoA . It is proposed that these reactions are part of the mechanism which enables certain acetogenic autotrophic bacteria to grow on CO2 and H2. J Clin Microbiol, 1984 Jan, 19(1), 60 - 2 Measurement of microbial alpha-amylases with p-nitrophenyl glycosides as the substrate complex; Trepeta RW et al.; The detection of alpha-amylase is commonly used in clinical microbiology laboratories to aid in differentiating Streptococcus bovis from other streptococci . It is also useful in identifying Eikenella corrodens and the gravis subspecies of Corynebacterium diphtheriae and in separating species of the genera Bacteroides, Clostridium, Actinomyces, and Bacillus . Currently, the most frequently used procedure utilizes starch as the substrate and iodine as the indicator . Starch is incorporated into a agar medium, the isolate is inoculated on the surface, and the medium is incubated for 24 to 48 h . A 15-min test containing p-nitrophenyl polyglycosides as the substrate complex was developed to yield results comparable with the agar-based starch test . The reagent was made in liquid form, 0.20 ml per tube, and could be incubated either in ambient air or at 35 degrees C . When dried, the p-nitrophenyl polyglycoside reagent could be stored at 0 degrees C for 4 weeks. Boll Ist Sieroter Milan, 1984, 63(6), 552 - 5 {Food poisoning caused by Clostridium botulinum type E}; Aureli P et al.; The results of a microbiological investigation carried out into a home-canned tuna fish are reported in relation to a suspected botulism case . Toxin of Cl . botulinum type E was detected by mouse toxicity and neutralization tests . The food specimen were also cultured for Cl . botulinum . The isolates was identified as Cl . botulinum type E by biochemical, gas chromatographic and immunological tests . The outbreak in which for the first time in Italy, the Cl . botulinum type E is involved, concerns one person who showed typical signs and symptoms consistent with botulism (abdominal cramps, dilatated pupils, diplopia, dysphagia, paralysis of lower upper limbs) . The laboratory results are discussed with relation to environmental characteristics of the micro-organism and their resistance to same chemical and physical factors with are involved in the canning practice. Int J Gynecol Pathol, 1984, 3(4), 348 - 60 Isolation and ultrastructure of rabbit ovarian mesothelium (surface epithelium); Nicosia SV et al.; This study evaluated the efficacy of various dissociating procedures for isolating mesothelial or surface epithelial cells from rabbit ovaries . The procedure which provided the highest and most homogenous cell yield involved the following sequence: (a) incubation of whole ovaries for 60 min in Clostridium histolyticum collagenase, type I (300 U/ml); (b) vortexing of ovaries at 30 rpm for 60 s in medium 199; (c) removal of partially detached surface epithelium by gentle scraping with a microdissecting blade; and (d) unit gravity enrichment of isolated epithelial organoids (surface layers and villous processes) . As shown by light and ultrastructural microscopy, this procedure cleaved the surface epithelium from over two-thirds of the underlying basement membrane and tunica albuginea . As estimated by further dispersal of epithelial organoids with trypsin-EDTA, 0.8 to 1.5 X 10(6) surface cells were routinely obtained from a single ovary . This isolation procedure is simple and provides an experimental tool for investigating in vitro the regulation and developmental behavior of ovarian surface epithelium. Prog Clin Biol Res, 1984, 152, 163 - 74 Bacteriology of hepatolithiasis; Tabata M et al.; Hepatolithiasis is associated with bile stasis and bacterial infection . Gallstones found in the intrahepatic bile duct are mostly calcium bilirubinate stones, the presence of which is closely related to the presence of bacteria . In the present study, a high incidence of bile infection was found in hepatolithiasis: 52 of 54 cases (96.3%) . This is in concordance with the other reports from Japan as well as from East Asia . E coli was the most frequent isolate followed by Klebsiella, Streptococcus (D), and Pseudomonas . Because of the frequent isolation of E coli in calcium bilirubinate stone cases, beta-glucuronidase from E coli has been thought to be responsible for the formation of calcium bilirubinate stones by effecting hydrolysis of bilirubin glucuronide to free bilirubin, which is insoluble in water . The recent introduction of improved anaerobic culture techniques has led to an increasing number of reports on the presence of anaerobes in the biliary tract . Anaerobes were isolated in 6 of 29 cases of hepatolithiasis (20.7%) in our series but more frequently in Kaohsiung, Taiwan (25 of 57 cases, or 44.4%) . Bacteroides and Clostridium were the most frequent isolates from the biliary tract and were shown to have beta-glucuronidase activity . Anaerobes were often found together with aerobes, suggesting the possibility of a synergistic effect that may influence the occurrence and development of cholangitis, which is often associated with hepatolithiasis . Though the biliary tract and liver are usually sterile, when an infection of the biliary tract occurs the route by which bacteria reach the region is thought to be hematogenous, lymphatic, or direct intraluminal ascending infection, the last being the most likely . Treatment of cholangitis associated with hepatolithiasis should be directed toward the removal of stones and termination of bile stasis . When cholangitis ensues, control of bacterial infection by antibiotics should be started without delay . The choice of antibiotics in controlling cholangitis is presented. Scand J Gastroenterol Suppl, 1984, 91, 31 - 43 Assay of metronidazole by HPLC compared with microbial method; Salvesen B et al.; A high-performance liquid chromatography (HPLC) method for the assay of metronidazole and its 2-hydroxymethyl metabolite in sera was compared with a microbiological method, an agar well diffusion technique with Clostridium perfringens as indicator strain . The HPLC technique involves separation of metronidazole from its two active major metabolites (the 2-hydroxymethyl and the 1-carboxymethyl derivatives) on a mu-Bondapak C18 column and UV detection at 313 nm . The mobile phase was 35% acetonitrile in 0.02 M acetate buffer pH 4 with a low rate 2.0 ml/min . Tinidazole was used as an internal standard and metronidazole and its 2-hydroxymethyl metabolite quantitated by peak height ratios . The 1-carboxymethyl derivative was well separated from the other peaks . The HPLC procedure proved to be superior with respect to sensitivity (detection limits: 0-1 microgram/ml serum for both compounds), speed and precision . It discriminates between metronidazole and its two major active metabolites and quantitates the total amounts present . The microbial technique codetermines all antibacterial active compounds and monitors the free, not protein bound moieties . Published data on the activity of the 2-hydroxymethyl metabolite against Cl . perfringens relative to metronidazole and published results on the protein binding of metronidazole were used to correlate data from the two methods on individual serum samples collected during the early, intermediate and late periods after a single intravenous dose of metronidazole to volunteers. Scand J Gastroenterol Suppl, 1984, 91, 103 - 11 Evaluation of Minitec and API as rapid diagnostic methods for anaerobic bacteria; Bergan T et al.; The performance of the miniaturized biochemical differentiation systems API 20 anaerobe System and Minitek was compared with pH reactions in PRAS media of 58 anaerobic or capno-aerophilic bacterial strains from recent clinical specimens . The overall correlation of the Minitek reactions was 87% and of the API reactions 85% . The reactions of acid formation from carbohydrates was slightly higher for Minitek . This resulted in a correct diagnosis in 90% after Minitek and 86% after API . If corrected for 3 strains of Clostridium perfringens for which double haemolysis would have prevented reporting of another diagnosis, 95% of the strains would have been correctly identified by Minitek compared to 91% with API . The errors were mostly due to lack of sensitivity of the reactions of the test kits. Adv Intern Med, 1984, 29, 85 - 107 Clostridium difficile colitis; Trnka YM et al.; Clostridium difficile has become one of the commonest pathogens of the lower intestinal tract . This organism appears unique in that infection almost always occurs during or after antibiotic therapy, suggesting that some component of the normal microflora prevents colonization by C . difficile . Once it has overgrown in the colon, C . difficile releases several toxins which cause tissue damage and diarrhea . Infection can range from a simple self-limited diarrheal illness to fulminant colitis with perforation and megacolon . Assay of stool filtrates reveals the presence of cytotoxin in nearly all patients with antibiotic-associated pseudomembranous colitis, and in approximately one third to one half of those with less severe infections . Effective therapy is available in the form of oral vancomycin, although the expense of this antibiotic has led to the use of oral metronidazole or bacitracin, which appear to be equally efficacious and considerably cheaper . Although we have learned a great deal about C . difficile in the past decade, a number of fascinating puzzles remain . We know very little about the immune response to this organism or its toxin, or whether a vaccine might someday be feasible . Similarly, we have very little insight into what effects antibodies exert on the normal colonic flora and how these effects allow C . difficile infection in a small percentage of patients . Studies of this pathogen will undoubtedly lead to a fuller understanding of the enormously complex and still mysterious microbial ferment which lives within our gastrointestinal tract. Appl Environ Microbiol, 1984 Jan, 47(1), 28 - 30 Quantitation of pH- and salt-tolerant subpopulations from Clostridium botulinum; Montville TJ; Plating efficiencies of Clostridium botulinum 62A spores on media with variable pH (7.0 to 5.5) and salt (0, 1, 2, and 3%) levels revealed that only a very small subpopulation could give rise to colonies . The relative size of this subpopulation decreased by orders of magnitude with decreasing pH and increasing salt concentrations . Strong interactions of pH with salt were noted . For example, on a medium containing 2% salt at pH 5.5, colonies could be formed from only 1 in 100,000 spores . Proper monitoring of medium anaerobiosis was critical in obtaining reproducible results. Infect Immun, 1984 Jan, 43(1), 6 - 10 Bacterial translocation from the gastrointestinal tracts of rats receiving thermal injury; Maejima K et al.; Rats receiving nonlethal thermal burns over 20 or 40% of their total body surface area were tested at various intervals for the translocation of indigenous bacteria from their gastrointestinal tracts to their mesenteric lymph nodes, peritoneal cavities, and bloodstreams . No indigenous bacteria were cultured from these organs of control rats or from rats receiving 20% burns . However, 44% of the rats receiving 40% burns exhibited viable Escherichia coli, Proteus mirabilis, Staphylococcus sp . and Clostridium sp . in their mesenteric lymph nodes 2 days after thermal injury . Bacterial translocation after burn stress also was tested in antibiotic-decontaminated rats monoassociated with E . coli . E . coli attained population levels in these animals of 10(8) to 10(9) per g cecum . E . coli translocated to 100% of the mesenteric lymph nodes of both the control and 40% burned rats . However, E . coli translocated at a greater incidence to the spleens, livers, and peritoneal cavities of the burned rats compared with translocation to these organs in control rats . The numbers of E . coli translocating to the mesenteric lymph nodes, spleens, and livers also were greater in the 40% burned rats than in control rats . By 14 days after thermal injury, the rats were able to clear E . coli from their spleens and livers, and the infection remained localized in the mesenteric lymph nodes . These results support the concept that the indigenous gastrointestinal flora or exogenous organisms colonizing the gastrointestinal tract are potential sources of septicemia after thermal injury. Scand J Infect Dis Suppl, 1984, 42, 72 - 82 Anaerobic bacteria and beta-lactam antibiotics; Nord CE et al.; This review is concerned with anaerobic bacteria and beta-lactam antibiotics . Data on the susceptibility to beta-lactam antibiotics of anaerobic isolates in Sweden during 1983 is presented . Among the different bacterial groups, Clostridium perfringens, anaerobic cocci and curved rods were found to be very susceptible to the beta-lactams tested . C . difficile was susceptible to benzylpenicillin, piperacillin and imipenem, but resistant to the second and third generation of cephalosporins . Most fusobacteria were susceptible to the beta-lactam antibiotics investigated . However, some strains were found to be resistant to the penicillins . Against the Bacteroides fragilis group, imipenem had the best activity whereas moxalactam, piperacillin and cefoxitin had good activities . The majority of the Bacteroides non-fragilis strains were still susceptible to beta-lactam antibiotics . However, an increasing number of isolates were found to be resistant, especially to the penicillins when comparing with data from the last five years . The known mechanisms of beta-lactam resistance in anaerobic bacteria involve production of beta-lactamases, alteration of penicillin-binding proteins and blocked penetration of beta-lactams through the outer membranes of anaerobes . The presence of beta-lactamases in various Bacteroides and Clostridium species is reported as well as the characteristics of the different beta-lactamases encountered . The role of penicillin-binding proteins and penetration barrier in beta-lactam resistance is also discussed. Microbiol Immunol, 1984, 28(3), 281 - 9 Interrelationship between drug resistance and bacteriocinogeny of Clostridium perfringens; Miyoshi Y et al.; One hundred and eight strains of Clostridium perfringens were collected from polluted waters and tested for their drug resistance and bacteriocinogeny . Thirty of the strains were tetracycline resistant and 36 were bacteriocinogenic . Only one strain possessed both tetracycline resistance and bacteriocinogen . Transfer experiments on tetracycline resistance and bacteriocinogeny were carried out with several selected strains among these isolates . Both tetracycline resistance and bacteriocinogeny were shown to be transferable by conjugation-like procedures . Transfer experiments on these two properties revealed that there were two types of recipient strains among the isolates in regard to the acceptance of tetracycline resistance and bacteriocinogeny . In one type of the recipient strain, these two properties seemed to be incompatible although in the other, both tetracycline resistance and bacteriocinogeny were transferred by two-step mating . Transcipients of this two-step mating retransferred their tetracycline resistance but not bacteriocinogeny to another recipient strain . The existence of an incompatibility-like relationship between tetracycline resistance and bacteriocinogeny in one recipient strain, and the inability of these two properties to be co-transferred from the two-step transcipients , suggest a certain interrelationship between the factors responsible for tetracycline resistance and bacteriocinogeny . Arzneimittelforschung, 1984, 34(3), 321 - 5 Ceftriaxone: therapeutic results in various infections and kinetic studies; Giamarellou HJ et al.; Ceftriaxone (Ro 13-9904, Rocephin) was given to 67 patients suffering from 74 various infections . Patients had infections of the urinary tract (36), soft tissue phlegmon (12), infections of the respiratory tract (13), osteomyelitis (7), abscesses (5) and meningitis (1) . Infecting organisms were E . coli (26), Proteus spp . (20), P . aeruginosa (7), H . influenzae (6), Enterobacter spp . (6), K . pneumoniae (3), C . freundii (2), S . marcescens (1), S . aureus (4), S . pneumoniae (2), Peptostreptococcus spp . (4) and Clostridium spp . (1) . The organisms were often multiresistant . Dosage ranged from 1-2 g once or twice daily i.m . or i.v . The clinical response was excellent in 56 infections (75.6%) while 13 (17.6%) infections were improved and in 5 (6.8%) treatment failed . The pathogen was eradicated in 63 (85.1%), and relapsed in 3 (4.0%) while bacteria persisted in 11 (14.9%) . No appreciable side effects or toxicity were observed . Sputum, bile, cerebrospinal fluid and prostatic fluid kinetics revealed ceftriaxone concentrations several times above the minimal bactericidal concentrations required to kill the enterobacteriaceae, but mostly inadequate to inhibit P . aeruginosa strains . Ceftriaxone was a safe and effective drug for the treatment of various infections . However, with the exception of urinary tract infections it should be given with caution in P . aeruginosa infections. Zentralbl Bakteriol Mikrobiol Hyg {B}, 1984 Jan, 178(5-6), 542 - 50 {Effect of heating beef at 55 degrees C on the growth of Clostridium perfringens and Staphylococcus aureus}; Labadie J et al.; A tenderizing method has been set up for beef meat by heating be at 55 degrees C . The present paper deals with informations about the pieces of meat which could treated by this method and particularly about the growth of Clostridium perfringens and Staphylococcus aureus . Our results show that pieces of meat the dimensions of which are 8.8.8 . cm seem to be usable without danger for the consumers. Zh Mikrobiol Epidemiol Immunobiol, 1984 Jan, (1), 42 - 6 {Pathology and physiology of microbes . III . Criteria for assessing functional states}; Mel'nikova VA et al.; The study of different characteristics of the vital activity of microorganisms, made on type B Clostridium perfringens taken as an example, has disclosed the difficulty of evaluating the functional state of microorganisms only on the basis of the traditionally used characteristics of the population activity: the specific growth rate, the toxicity of the culture fluid and the antigenic properties of the culture . The economic and metabolic coefficients have been found to be the most suitable criteria for such evaluation; the state of the maximum physiological activity of the population is characterized by the maximum values of the economic coefficient and the minimum values of the metabolic coefficient . The minimum values of the economic coefficient and the maximum values of the metabolic coefficient are characteristic of the pathological state of the population. Exp Cell Res, 1984 Jan, 150(1), 194 - 204 Preparation of isolated liver endothelial cells and Kupffer cells in high yield by means of an enterotoxin; Blomhoff R et al.; A new method for preparing non-parenchymal rat liver cells (NPC) is described . The liver cell suspension, prepared by perfusing the liver with collagenase, was treated with enterotoxin from Clostridium perfringens for 15 min . The enterotoxin made the parenchymal cells leaky, and these cells could be separated from the NPC by centrifugation in a solution containing Nycodenz (20%, w/v) . During the centrifugation, the NPC floated, while the parenchymal cells sedimented . The yield of NPC per liver (200 g rat) was about 250 X 10(6) cells . The NPC were further separated into endothelial cells, Kupffer cells and stellate cells by centrifugal elutriation . This method was particularly useful for preparing endothelial cells in high yield (100 X 10(6) cells per liver) . Intravenously injected formaldehyde-treated albumin was selectively taken up by the endothelial cells . Isolated endothelial cells in suspension as well as in surface culture maintained their ability to endocytose this ligand. Infect Immun, 1984 Jan, 43(1), 54 - 8 Histopathological effect of botulinum C2 toxin on mouse intestines; Ohishi I et al.; Botulinum C2 toxin has histopathological activity in the mouse intestine and induces fluid accumulation in intestinal loops . The toxin caused degenerative and necrotic changes in the intestinal mucosa: intracellular vacuolization of epithelial cells, desquamation and necrosis of the villous epithelium, intercellular edema, and infiltration of lymphocytes and histiocytes . The detectable changes in the morphology of the intestinal mucosa preceded the increase in fluid accumulation in intestinal loops . Intraluminal injection of botulinum C2 toxin also induced the leakage of plasma protein into the intestinal lumen as determined by the extravasation of Evans blue . In contrast to botulinum C2 toxin, cholera and Clostridium perfringens enterotoxin controls caused a very slight protein leakage, although these toxins induced marked fluid accumulation in intestinal loops . The results indicate that the mode of action of botulinum C2 toxin in eliciting the secretory response is distinguishable from those of cholera and C . perfringens enterotoxins and suggest that botulinum C2 toxin induces the secretory response by cytopathic effect(s) on the epithelial cells of the intestine. Microbiol Immunol, 1984, 28(1), 23 - 31 Effect of Clostridium perfringens beta toxin on blood pressure of rats; Sakurai J et al.; Guanethidine treatment or adrenal medullectomy significantly inhibited the elevation in blood pressure induced by Clostridium perfringens beta toxin, and the combination of the two drastically reduced the pressure rise, to less than 19% of that in control rats . When rats were pretreated with tetrodotoxin or hexamethonium, the toxin-evoked rise was significantly inhibited . Elevation in blood pressure induced by the toxin in spinal rats tended to be less than that in control rats . When investigated by a microscopical technique, arteriolar constriction in the mesenteric vasculature was observed after the blood pressure elevation induced by the toxin reached a maximum . Blood flow in the skin decreased with an increase in blood pressure following intravenous injection of the toxin . It is concluded that beta toxin acts on the autonomic nervous system and produces arterial constriction. Rev Argent Microbiol, 1984, 16(1), 33 - 8 {Enzyme immunoassay in the detection of Clostridium perfringens enterotoxin}; Dobosch D; An enzyme-linked immunosorbent assay (ELISA) for C . perfringens enterotoxin detection was evaluated using purified enterotoxin preparations, supernatant fluids of sporulating C . perfringens cultures and other Clostridium species . The assay was found satisfactorily sensitive and specific to detect C . perfringens enterotoxin. Scand J Infect Dis Suppl, 1984, 43, 44 - 9 Antimicrobial susceptibility of anaerobic bacteria in Sweden in 1983; Nord CE et al.; This review is concerned with anaerobic bacteria and antimicrobial susceptibility . Data on the susceptibility to different antimicrobials of anaerobic isolates in Sweden during 1983 is presented . Among the different bacterial groups, Clostridium perfringens, propionibacteria, anaerobic cocci and curved rods were found to be susceptible to beta-lactam antibiotics, clindamycin and chloramphenicol . C . difficile was susceptible to benzylpenicillin and imipenem, but resistant to cefoxitin . Most C . difficile strains were also susceptible to clindamycin and fusidic acid, while all strains were susceptible to metronidazole and vancomycin . The majority of the fusobacteria were susceptible to beta-lactam antibiotics, clindamycin, metronidazole and chloramphenicol . However, some strains were found to be resistant to penicillins . Against the Bacteroides fragilis group, clindamycin, chloramphenicol, metronidazole and imipenem had the best activity, whereas piperacillin and cefoxitin showed good activities . Most Bacteroides non-fragilis strains were susceptible to the antimicrobials tested . However, an increasing number of isolates were found to be resistant especially to penicillins when comparing with data from the last five years. Respiration, 1984, 46(4), 382 - 5 Spontaneous bacterial pleuritis in a patient with cirrhosis; Streifler J et al.; Empyema of the left pleural cavity developed suddenly in a nonalcoholic cirrhotic patient . Cultures of the pleural fluid under anaerobic conditions grew Clostridium perfringens, an organism normally found in the enteric flora . The infection developed in an old pleural effusion . Since there was no evidence of trauma, necrotizing pneumonitis or subphrenic infection, spontaneous bacterial pleuritis is proposed. Vet Med Nauki, 1984, 21(6), 34 - 9 {Anaerobic dysentery in pigs}; Ganovski D et al.; Anaerobic dysentery (necrotic enteritis) of sucking pigs was demonstrated on an industrial swine-breeding farm . The disease was found to be caused by Clostridium perfringens, type C, and attacked up to 3 per cent of all sucking pigs at the age of 1 to 5 days, mortality rate ranging as high as 50 per cent . Outbreaks were recorded only in boxes fixed on the floor of the premise itself . The control of the disease made use of complex measures, including washing of sows with disinfection solution prior to their accomodation in the farrowing unit, strict observation of the principle 'all in, all out', feeding medicated premixes to the sows, improved hygiene on the premise, vaccination of the sows with an antiperfringens vaccine, and oral treatment of the diseased pigs prior to sucking with a specific hyperimmune antitoxic serum as well as using broad-spectrum antibiotics, spectinomycin, and pharmazin . With all measures applied the disease was fully suppressed. Biochem Biophys Res Commun, 1983 Dec 28, 117(3), 803 - 8 Stimulation of prothrombinase activity of platelets and erythrocytes by sub-lytic treatment with phospholipase C from Clostridium welchii; Comfurius P et al.; Treatment of platelets or red cells with small amounts of phospholipase C from Clostridium welchii enables both cells, prior to the onset of lysis, to stimulate prothrombin conversion by coagulation factor Xa and Va in the presence of calcium . Phospholipase C treatment of both cells also exposes significant amounts of phosphatidylserine at the outer surface . The level of phosphatidic acid formed from diglycerides produced by phospholipase C action, is similar to that formed in activated platelets upon triggering the phosphatidylinositol cycle . A possible involvement of this cycle to activate platelets to become more procoagulant is discussed. Biochim Biophys Acta, 1983 Dec 19, 763(4), 383 - 92 Internalization of Clostridium difficile cytotoxin into cultured human lung fibroblasts; Florin I et al.; In cultured human lung fibroblasts treated with Clostridium difficile cytotoxin, the latency before appearance of the cytopathogenic effect was dose-related with a minimum of 45 min . At 37 degrees C, the toxin was accessible on all cells to inactivation with trypsin or neutralization with antitoxin during the first tenth of the latency . At 0 degrees C, the toxin was accessible considerably longer . The cytopathogenic effect was reversibly prevented by the lysosomotropic agents chloroquine and ammonium chloride, which had to be added within one-fifth of the latency to protect all cells . In the presence of chloroquine, but not of ammonium chloride, the time period during which the toxin remained amenable to neutralization with antitoxin was prolonged . The protective effect of ammonium chloride was not influenced by dropping the extracellular pH to 4.5, but that of chloroquine was abolished . The expression of the intoxication was not affected by inhibitors of the DNA, RNA or protein synthesis . Inhibitors of the energy metabolism prevented the cytopathogenic effect when added before the last phase of the latency . The results suggest that expression of the cytopathogenic effect requires internalization of the toxin, and that metabolic energy but no macromolecular synthesis is needed for the action of the toxin after this internalization. Appl Biochem Biotechnol, 1983 Dec, 8(6), 491 - 503 Production of acetic acid by immobilized whole cells of Clostridium thermoaceticum; Wang G et al.; Immobilized cells of Clostridium thermoaceticum for acetic acid production has been investigated . Using kappa-carrageenan gel as the immobilization-matrix, high cell concentration within the gel could be achieved and thus lead to high volumetric acetic acid productivity . Batch experiments using 3% gel showed that cell concentration up to 65 g (dry cell weight)/L gel could be achieved . These dry weight cell concentrations in the gel through immobilization are typically 10-15 times greater than what can be obtained in free-cell fermentations . The specific growth rate and acetic acid formation rate were similar to those observed for the free cells . Continuous culture experiments using a feed medium containing 20 g/L of glucose were performed where the reactor contained 50% by volume of the carrageenan gel and the pH was controlled at 6.9 . Different steady states were acheived at dilution rates ranging from 0.061 to 0.399 h-1 . Cells grew mainly near the surface of the gel and reached maximum concentration within the matrix of approximately 35 g/L . Dilution rates much greater than the maximum specific growth rate were obtained, which resulted in volumetric productivity up to 4.9 g/L-h . This value was significantly greater than that for the conventional continuous culture with free cells . Using a 40 g/L feed glucose concentration, steady states could be achieved between dilution rates of 0.12-0.4 h-1 . The maximum productivity further increased to 6.9 g/L-h at a dilution rate of 0.37 h-1 and at an acetic acid concentration of 19 g/L . The cell concentration was 60 g (dry weight)/L gel at steady state. Acta Pathol Microbiol Immunol Scand {B}, 1983 Dec, 91(6), 395 - 400 Toxin A of Clostridium difficile: production, purification and effect in mouse intestine; Lonnroth I et al.; Clostridium difficile produces one diarrhoeogenic toxin designed A, and one cytopathogenic toxin designed B . Toxin A was purified in a four-step-fractionation procedure . In the last purification step the toxin was separated by elution with galactose from an agarose gel . The purified toxin A induced a clear and watery hypersecretion in intestinal loops of mouse, while mixtures of toxin A and B induced a haemorrhagic secretion . At an ED50 value for the purified toxin A of 0.5 microgram there was a brief, optimal hypersecretion after four hours . Like the fluid secretion induced by cholera toxin, that induced by toxin A could be inhibited by chlorpromazine or by depletion of intestinal bile . In contrast to cholera toxin, however, toxin A did not activate intestinal adenylate cyclase--at least not permanently . Antisera which neutralized cholera toxin did not neutralize toxin A, and vice versa. Diagn Microbiol Infect Dis, 1983 Dec, 1(4), 331 - 3 Detection of Clostridium difficile cytotoxin in HEp-2 and CHO cell lines; Murray PR et al.; The detection of Clostridium difficile cytotoxin was compared in two established cell lines; Chinese hamster ovary (CHO) and a human epithelial line, HEp-2 . All specimens with positive toxin assays were detected with the CHO cell line, whereas only one-half of positive specimens were detected with the HEp-2 cells. Infect Immun, 1983 Dec, 42(3), 986 - 9 Infectivity of organisms recovered from polymicrobial abscesses; Brook I et al.; The ability to cause subcutaneous abscesses in mice was used to identify the pathogens among the bacteria recovered from 13 clinical abscesses . A total of 35 isolates (30 anaerobes and 5 aerobes), 16 of which were encapsulated, were recovered from these abscesses . Encapsulated organisms included eight Bacteroides spp . (three Bacteroides asaccharolyticus and one strain each of Bacteroides oralis, Bacteroides intermedius, Bacteroides biacutus, Bacteroides vulgatus, and Bacteroides ruminicola subsp . brevis), three anaerobic gram-positive cocci, two Clostridium spp., and two strains of Escherichia coli . Single organisms, or combinations thereof, obtained from these abscesses were inoculated subcutaneously into mice . All but one of the encapsulated organisms were able to cause abscesses by themselves and were recovered from the abscesses when inoculated with other organisms . Seven non-encapsulated organisms which were also recovered mixed with the encapsulated organisms were never able to cause abscesses nor could they be recovered after they were injected alone . Thirteen strains that did not induce abscesses when injected by themselves into mice survived when injected with other organisms which were encapsulated . In three instances, pairs of non-encapsulated organisms belonging to the last group were able to induce an abscess and survive in it when inoculated together . It was shown that the possession of a capsule by a clinical isolate increases the likelihood that it is a major contributor to the infectious process. Am J Public Health, 1983 Dec, 73(12), 1385 - 8 Infant botulism in the United States: an epidemiologic study of cases occurring outside of California; Morris JG Jr et al.; Data were obtained for the 96 hospitalized cases of infant botulism reported to the Centers for Disease Control between 1976-1980 from all states other than California . Forty-one cases were associated with Clostridium botulinum type A, 53 with type B, one with type F, and one with a strain of C . botulinum capable of producing both type B and F toxin . Cases occurred in 25 states; the disease was more common in the western part of the United States, with the highest attack rates reported for Utah and New Mexico . Birth-weights of hospitalized infants with infant botulism tended to be high compared with birth-weights in the United States population . Mothers of infants with infant botulism tended to be older and better educated than mothers in the general population . Seventy per cent of infants had been predominantly breast-fed; breast-feeding in type B cases was associated with a significantly older age at onset of illness. Anal Biochem, 1983 Dec, 135(2), 349 - 54 Estimation of ursodeoxycholic acid in human and bear biles using Clostridium absonum 7 beta-hydroxysteroid dehydrogenase; MacDonald IA et al.; Ursodeoxycholic acid was estimated in bile samples from humans and wild North American black bears using 7 beta-hydroxysteroid dehydrogenase purified from Clostridium absonum by Procion Red affinity chromatography . The percentage ursodeoxycholic acid was calculated by two methods: (a) 7 beta-hydroxyl groups were quantified using 7 beta-hydroxysteroid dehydrogenase and 3 alpha-hydroxyl groups (total bile acids) were quantified using 3 alpha-hydroxysteroid dehydrogenase . The percentage ursodeoxycholic acid was calculated on the basis of {7 beta-hydroxyl groups}/{3 alpha-hydroxyl groups} X 100 . (b) Bile was hydrolyzed with sodium hydroxide and subjected to thin-layer chromatography . Bands corresponding to cholic acid, chenodeoxycholic acid plus deoxycholic acid, and ursodeoxycholic acid were identified by the use of standards and Komarowsky's spray reagent . Total bile acids and total ursodeoxycholic acid were measured by elution of silica gel in unsprayed areas corresponding to the bile acid standards and quantification of the total bile acid in each eluate . Direct comparison of these methods validated the use of 7 beta-hydroxysteroid dehydrogenase in the estimation of ursodeoxycholic acid in the biles of black bears and of patients fed ursodeoxycholic acid for cholesterol gallstone dissolution . Relative percentages of ursodeoxycholic acid were 8-24% in four bears and 22 and 27% in the patients ingesting 500 and 750 mg ursodeoxycholic acid per day for 3 months, respectively . Predictably lower values were obtained in two control subjects and one patient ingesting 750 mg chenodeoxycholic acid per day for 3 months. Aust Vet J, 1983 Dec, 60(12), 374 - 7 Type C botulism in young dogs; Farrow BR et al.; A diffuse lower motor neurone paralysis developed in a 6-month-old male Australian cattle dog pup 4 days after it had eaten the carcase of a rotting duck in Centennial Park, Sydney . Two other dogs which ate smaller portions of the same carcase were less severely affected . Clostridium botulinum type C was isolated from and C . botulinum type C toxin was detected in faeces from the severely affected dog . The serum contained 25 LD50 of toxin/ml . The high C . botulinum count and toxin level in the faeces declined progressively during the ensuing weeks, but 114 days after ingesting the carcase C . botulinum type C was still present in faeces and a low toxin titre persisted . Soil, mud and water samples in the area of the duck ponds in the park contained C . botulinum type C spores . Spores and high toxin titres were also found in the intestine of the carcases of 2 birds in the area. Appl Environ Microbiol, 1983 Dec, 46(6), 1450 - 2 Failure of nisin to inhibit outgrowth of Clostridium botulinum in a model cured meat system; Rayman K et al.; Up to 550 ppm (550 micrograms/ml) of nisin in combination with 60 ppm (60 micrograms/ml) of nitrite failed to prevent outgrowth of Clostridium botulinum spores in pork slurries adjusted to pH 5.8 . Reducing the pH enhanced nisin activity . Proteolytic and nonproteolytic type B spores were equally resistant to nisin. J Clin Microbiol, 1983 Dec, 18(6), 1378 - 83 Fluorescent-antibody reagents for the identification of Clostridium botulinum; Glasby C et al.; Fluorescent-antibody reagents were prepared against vegetative cells of representative strains of each physiological group and toxin type of Clostridium botulinum known to have caused botulism in humans . A fluorescent-antibody reagent was also prepared for C . botulinum type G, which has been isolated from autopsy specimens but which has not clearly been implicated in botulism . These fluorescent-antibody reagents were evaluated against 200 strains of C . botulinum and 64 strains of other clostridia . Each reagent reacted with at least a 2+ intensity with all of the strains in its same toxin type and physiological group . Ninety-seven percent of the strains gave at least a 3+ reaction with the homologous group or toxin type reagent . Some cross-reactions occurred with reagents against different toxin type strains within a physiological group; there was less cross-reaction between physiological groups and very little reactivity of C . botulinum reagents with nontoxigenic organisms . Absorption of cross-reacting antibodies was not successful . Certain reagents could be used for presumptive laboratory identification of C . botulinum strains causing botulism, especially in infants . The type G reagent provided a good means of identifying C . botulinum type G, which lacks the lipase marker and whose toxigenicity may be more difficult to demonstrate in mixed cultures . There was a serological relationship between C . botulinum type G and some strains of Clostridium subterminale . This relationship provided evidence of differences between strains of C . botulinum type G isolated in two different countries. Int J Zoonoses, 1983 Dec, 10(2), 105 - 10 Use of enzyme linked immunosorbent assay (ELISA) in the quantitation of Clostridium perfringens type A enterotoxin and antienterotoxin antibodies; Narayan KG et al.; ELISA procedure for quantitation of enterotoxin of Clostridium perfringens type A was worked out . It was possible to detect as little as 0.0022 microgramme of enterotoxin . When applied to quantitate the enterotoxin in laboratory contaminated faeces and cooked minced beef, ELISA detected as little as 0.024 microgramme of enterotoxin . ELISA procedure for determination of titre of antibodies to enterotoxin was also developed using hyperimmunised rabbit and sheep sera . It was possible to use this procedure in sero-survey. Hoppe Seylers Z Physiol Chem, 1983 Dec, 364(12), 1653 - 63 On nitroaryl reductase activities in several Clostridia; Angermaier L et al.; Crude extracts of Clostridium kluyveri, Clostridium spec . La 1, Clostridium sporogenes and Clostridium pasteurianum catalyse the NADH-dependent reduction of the nitro group of p-nitrobenzoate . The former three Clostridia also use pyruvate as electron donor for this reduction . The NADH-dependent reductases have been partially purified and characterized from Clostridium kluyveri . Nitroalkyl compounds as well as nitrite, sulfite, sulfate and hydroxylamine are no substrates . Based on chromatographic behavior, separation pattern, yields, stability, pH optima, molecular masses and EPR studies the three NADH-dependent nitroaryl group reducing enzymes in Clostridium kluyveri (three activities in Clostridium spec . La 1 and two activities in Clostridium sporogenes) are different from alcohol dehydrogenase, aldehyde dehydrogenase, 3-hydroxy-butyryl-CoA dehydrogenase, butyryrl-CoA dehydrogenase, 2-enoate reductase, ferredoxin-NAD and ferredoxin-NADP reductase . The physiological roles of the nitroaryl reductases are not known . The reductase activities show losses of 80-90% during classical protein purification procedures . One of the three nitroaryl reductases exhibits a pH optimum of 10.5 . The crude extract reveals a pH optimum at 11.5 . The first step of the reduction reaction leads to the nitroradical anion (1 electron transfer) . The electron transfer to p-nitrobenzoate is also catalysed by ferrodoxin-NAD reductase from NADH and by ferredoxin-NADP reductase from NADP . Partially purified 2-oxo-acid synthases from Clostridium sporogenes catalyse with low rates the reduction of p-nitrobenzoate as well as 2-nitroethanol in the presence and absence of ferredoxin using pyruvate or 2-oxo-4-methylpentanoate as electron donors, respectively . The NADH-dependent reduction of p-nitro-benzoate accounts for at least 70% and the 2-oxo acid-dependent reduction for about 5% of the total nitroaryl reductase activity in the Clostridia . It seems that the pyridine nucleotide-dependent nitroaryl reductases are enzymes so far unknown in Clostridia. Proc Natl Acad Sci U S A, 1983 Dec, 80(23), 7244 - 8 Phorbol esters, phospholipase C, and growth factors rapidly stimulate the phosphorylation of a Mr 80,000 protein in intact quiescent 3T3 cells; Rozengurt E et al.; Addition of biologically active phorbol esters to intact quiescent 3T3 mouse cells stimulates an extremely rapid (detectable within seconds) phosphorylation of a Mr 80,000 cellular protein (termed "80k") . Phorbol 12,13-dibutyrate enhances 80k phosphorylation in a dose-dependent manner; half-maximal effect is obtained at 32 nM . The possibility that this phosphorylation is related to the activation of Ca2+-activated phospholipid-dependent protein kinase is suggested by the fact that phospholipid breakdown induced by exogenous treatment of the cells with phospholipase C from Clostridium perfringens or with platelet-derived growth factor, which is a potent activator of endogenous phospholipase C activity, also causes a rapid enhancement of 80k phosphorylation . Moreover, prolonged pretreatment of the cells with phorbol 12,13-dibutyrate, which leads to a marked decrease in the number of specific phorbol ester binding sites, prevents the phosphorylation of 80k stimulated by phorbol esters, phospholipase C, and platelet-derived growth factor . These findings provide evidence obtained with intact cells that implicate the stimulation of Ca2+-activated phospholipid-dependent protein kinase in the action of phorbol esters and other growth factors. J Periodontol, 1983 Dec, 54(12), 740 - 5 Degradation of basement membrane collagen by proteinases from human gingiva, leukocytes and bacterial plaque; Uitto VJ; Basement membranes in human gingiva are found in dento-epithelial junction, epithelial-connective tissue junction and endothelium-connective-tissue junction where they have important attachment and filtering functions . The ability of plaque and gingiva-derived proteinases to degrade Type IV collagen, the major protein of basement membranes, was examined in vitro . The basement membrane collagen (Type IV) isolated from bovine lens capsules was incubated in the presence of enzyme samples . The degradation was assayed by the release of hydroxyproline from the insoluble substrate and by examining the peptide pattern of the residue by polyacrylamide gel electrophoresis . Salt extracts of inflamed gingival specimens degraded basement membrane collagen into soluble form and produced degradation products that were similar to those produced by human leukocyte extracts . Gingival crevicular fluid collected from patients with severe adult periodontitis also digested the substrate but the degradation pattern was different from the leukocyte and gingival extract samples . The pattern closely resembled the degradation produced by bacterial plaque extracts . A third type of cleavage was observed when collagenase from Clostridium histolyticum was incubated with basement membrane collagen . Crevicular fluid and the extracts from gingiva, leukocytes and plaque also contained gelatinase and elastase-like enzyme activities that have earlier been shown to be potent in degrading basement membrane . It was concluded that enzymes capable of degrading basement membrane collagen in gingivitis and periodontal disease may originate from both plaque bacteria and human leukocytes . It also appeared that the enzymes responsible are more likely to be gelatinase and elastase-like enzymes than specific collagenases. Infect Immun, 1983 Dec, 42(3), 1183 - 6 Effect of Clostridium perfringens epsilon toxin on the cardiovascular system of rats; Sakurai J et al.; Pressor activity was demonstrated by the intravenous injection of purified epsilon toxin of Clostridium perfringens type D but not by that of epsilon prototoxin . The activity was completely abolished by C . perfringens type B or D antiserum, but not by type A, C, or E antiserum . The rise in blood pressure caused by the toxin was accompanied by a decrease in the blood flow in skin without any change of heart rate and electrocardiogram readings . These results indicate that epsilon toxin possesses pressor activity as well as lethal and dermonecrotic activities. Gastroenterology, 1983 Dec, 85(6), 1403 - 6 Gastrointestinal cytomegalovirus infection in a homosexual man with severe acquired immunodeficiency syndrome; Gertler SL et al.; A 44-yr-old white homosexual man with a history of Pneumocystis carinii pneumonia developed watery diarrhea and fever . Flexible sigmoidoscopy revealed yellow plaques in the sigmoid colon suggestive of pseudomembranous colitis . Stool examination for ova and parasites, Clostridium difficile toxin, and cultures for pathogens, including Clostridium difficile, were negative . Infection with multiple organisms, including Mycobacterium avium intracellulare, Toxoplasma gondii, herpes simplex, and cytomegalovirus, was demonstrated . Postmortem examination showed esophageal and colonic ulcerations with cytomegalic inclusions, and cytomegalovirus was cultured from the colonic lesions . This report describes a case of disseminated cytomegalovirus infection with esophageal and colonic involvement in a homosexual man with the acquired immunodeficiency syndrome. J Hosp Infect, 1983 Dec, 4(4), 375 - 82 A prospective randomized trial to compare mezlocillin and metronidazole with cefuroxime and metronidazole as prophylaxis in elective colorectal operations; Ambrose NS et al.; A prospective randomized trial has compared a broad spectrum ureidopenicillin with a broad spectrum cephalosporin for prophylaxis against the aerobic organisms encountered during elective colonic surgery . Even though only two doses of antibiotics were administered the incidence of severe sepsis was low . Severe wound infection occurred in three of the patients receiving mezlocillin and metronidazole (6 per cent) compared with six in the group receiving cefuroxime and metronidazole (13 per cent) . Minor wound sepsis was recorded in 24 per cent of patients receiving mezlocillin and metronidazole compared with only 11 per cent after cefuroxime and metronidazole . There were two episodes of septicaemia, one in each group, and three abscesses, all of which occurred in patients receiving metronidazole and mezlocillin . The total number of surgically related infections was, however, significantly less with cefuroxime and metronidazole (N = 13) compared with mezlocillin and metronidazole (N = 23; P less than 0.03) . Escherichia coli was the principal organism responsible for surgically-related postoperative sepsis: (22 isolates: 14 mezlocillin and eight cefuroxime) all of which sensitive to the agents used . Pseudomonas aeruginosa was recovered from 10 patients (three mezlocillin and seven cefuroxime), all of the isolates were resistant to both antibiotics and were associated with severe morbidity . There were 11 isolates of Staphylococcus spp . (nine mezlocillin and two cefuroxime: P less than 0.03) . Postoperative diarrhoea occurred in six patients, all were in the group receiving cefuroxime and metronidazole . (Clostridium difficile was recovered from the stool in three of which one was associated with Cl . difficile cytotoxin.) FEBS Lett, 1983 Nov 14, 163(2), 212 - 6 Resonance Raman spectroscopy of Azotobacter vinelandii ferredoxin I . Vibrational features of the {3Fe-3S} cluster; Lutz M et al.; Low temperature resonance Raman spectra have been obtained for Clostridium pasteurianum and Bacillus stearothermophilus ferredoxins . Several heretofore undetected fundamental bands have been observed and these data have been used to discriminate the vibrational contribution of the {3Fe-3S} cluster to the spectrum of Azotobacter vinelandii ferredoxin I . The vibrational features of the {3Fe-3S} core distinguish it from other 3-iron clusters and imply structural differences among this class of iron-sulfur clusters. Lancet, 1983 Nov 5, 2(8358), 1043 - 6 Prospective randomised trial of metronidazole versus vancomycin for Clostridium-difficile-associated diarrhoea and colitis; Teasley DG et al.; 101 patients with Clostridium-difficile-associated diarrhoea or colitis were prospectively randomised to 10-day oral courses of metronidazole, 250 mg four times a day, or vancomycin, 500 mg four times a day . 7 did not complete the protocol and were dropped from analysis . Pseudomembranous colitis (PMC) was diagnosed after endoscopy in 33 patients . Of the remaining patients without PMC, 38 had both C difficile culture and cytotoxin and 23 had only culture evidence of C difficile . 52 evaluable patients received vancomycin and 42 received metronidazole . There were two treatment failures with metronidazole and none with vancomycin (p = 0.20); and two relapses with metronidazole versus six with vancomycin (p = 0.17) . Treatment in 1 patient in each group was discontinued because of drug intolerance . Response and relapse rates of the 33 patients with PMC were no different from those of the remaining patients . Pharmacy cost for the dosage used was $387.48 to $520.00 for vancomycin and $11.84 for metronidazole . Metronidazole and vancomycin have equivalent efficacy and relapse rates and are tolerated to a similar extent by patients with C-difficile-related diarrhoea and colitis, but metronidazole is considerably more economical. Eur J Biochem, 1983 Nov 2, 136(2), 427 - 34 Purification, characterisation and reconstitution of glutaconyl-CoA decarboxylase, a biotin-dependent sodium pump from anaerobic bacteria; Buckel W et al.; Glutaconyl-CoA decarboxylase from Acidaminococcus fermentans is a biotin enzyme, which is integrated into membranes . It is activated by Triton X-100 and inhibited by avidin . The results obtained by a combination of both agents indicate that biotin and the substrate-binding site are located on the same side of the membrane . The decarboxylase was solubilized with Triton X-100 and purified by affinity chromatography on monomeric avidin-Sepharose . The enzyme is composed of three types of polypeptides: the group of alpha chains (Mr 120000-140000) containing the biotin, the beta chain (60000) and an apparently hydrophobic gamma chain (35000) . Sodium ions specifically protected the latter chain from tryptic digestion . It was supposed, therefore, that this chain might function as the Na+ channel . The beta and gamma chains but not the alpha chain could be labelled by N-ethyl-{14C}maleimide . Similar decarboxylases but with much smaller biotin peptides (Mr 15000-20000) were isolated from Peptococcus aerogenes and Clostridium symbiosum . The decarboxylases from all three organisms could be reconstituted to active sodium pumps by incubation with phospholipid vesicles and octylglucoside followed by dilution . The Na+ uptake catalysed by the enzyme from A . fermentans was completely inhibited by monensin and activated twofold by valinomycin/K+ indicating an electrogenic Na+ pump . The coupling between Na+ transport and decarboxylation was not tight . During the reaction the ratio decreased from initially 1 to 0.2 . The three organisms mentioned above and Clostridium tetanomorphum without glutaconyl-CoA decarboxylase are able to ferment glutamate and require 10 mM Na+ for rapid growth . There is no correlation between the concentration of monensin necessary to inhibit growth and the presence of decarboxylase in these organisms. Klin Wochenschr, 1983 Nov 2, 61(21), 1081 - 7 {Clostridium difficile and antibiotic-associated colitis in risk patients: 2-month epidemiologic study in an intensive care unit}; Loeschke K et al.; A toxin produced by Clostridium difficile has been implicated in the pathogenesis of antibiotic-associated colitis . It is not known how often the microorganism is encountered in Germany particularly in high risk patients . Therefore, following a lethal case of colitis, stool samples of 90 patients and 30 staff members of an intensive care unit were screened routinely for C . difficile over 2 months . The organism was found in 6 of 41 patients treated with antibiotics (14.6%); four of them apparently acquired C . difficile while in hospital whereas in 2 a pre-existing carrier state could not be excluded . Colitis developed in 3 of the 6 patients as judged from endoscopy or a positive cytotoxin assay; in 2 patients (not subjected to endoscopy) colitis was suspected on clinical grounds, and 1 patient became an asymptomatic carrier . C . difficile was not found in 49 patients without antibiotic medication, in the health personal and in 12 patients of a general ward . Patients harbouring C . difficile were clustered in certain bed sites of the unit . Environmental studies recovered the microorganism from bed pan washing machines of bedridden and from toilets of ambulant patients but not from other sites like the hands of the personal . These results suggest that chronic carriers of C . difficile, as far as they are identified by current bacteriological methods, are rare in Germany (not more than 2 out of 132 persons investigated, i.e . 1.5%) . The frequent finding of C . difficile in patients treated in certain bed sites supports the view that the infection may be acquired from exogenous sources . Antibiotic-associated colitis should be considered more often when intensive care patients are treated with antibiotics. J Biochem (Tokyo), 1983 Nov, 94(5), 1715 - 8 Detection of D-erythro and L-threo sphingosine bases in preparative sphingosylphosphorylcholine and its N-acylated derivatives and some evidence of their different chemical configurations; Hara A et al.; Sphingosylphosphorylcholine prepared from native sphingomyelin by the Kaller procedure was found to comprise about 70% of the L-threo (2S, 3S) isomer and 30% of the D-erythro (2S, 3R) isomer . This analytical result was obtained by gas-liquid chromatography (GLC) of trimethylsilyl derivatives of N-acetylsphingosines which were prepared by enzymatic hydrolysis of synthetic N-acetylsphingosylphosphorylcholines with Clostridium perfringens phospholipase C . Some other evidence of the different chemical configuration between the erythro and threo isomers of synthetic N-acylated sphingosylphosphorylcholines was also provided by thin layer chromatography (TLC), optical rotatory dispersion (ORD), and fast atom bombardment (FAB) mass spectrometry. Appl Environ Microbiol, 1983 Nov, 46(5), 1103 - 12 Characterization of Lactobacillus sp . strain 100-37 from the murine gastrointestinal tract: ecology, plasmid content, and antagonistic activity toward Clostridium ramosum H1; McCormick EL et al.; A gram-positive, nonsporulating, microaerophilic rod that had two colonial variants was obtained during a study in which anaerobic bacteria were isolated from murine gastrointestinal tracts and screened for cryptic plasmids . The rod (both colonial variants) was identified as a Lactobacillus sp . (strain 100-37) by selective media, gas chromatography, and biochemical tests . In monoassociated, ex-germfree mice, the bacterium colonized the gastrointestinal tract and formed a thick, continuous layer on the keratinized squamous epithelium of the nonsecreting portion of the stomach . When lysate preparations of both colonial variants were electrophoresed in agarose gels, two bands which stained with ethidium bromide were detected with each lysate . When the DNA preparations were exposed to UV light, the lower ethidium bromide band gradually disappeared while the top band became either broader or more intense . The approximate size of the lower band was 2.2 megadaltons, as determined by comparison with plasmid molecular weight standards . In a search for phenotypes which could be encoded by the cryptic 2.2-megadalton plasmid, we detected an antagonistic activity toward an obligate anaerobe isolated from mouse feces, Clostridium ramosum H1 . The antagonistic factor was precipitated with (NH4)2SO4 (70% saturation) from supernatant solutions of broth cultures of strain 100-37 . The factor was not inducible with mitomycin C or UV light, but was stable in flowing steam for up to 50 min, and in buffers of pHs over a range of 1.6 to 6.8 . It was nondialyzable and inactivated by trypsin and papain.(ABSTRACT TRUNCATED AT 250 WORDS) J Assoc Off Anal Chem, 1983 Nov, 66(6), 1510 - 3 Freeze-dried mixed cultures as samples for proficiency tests and collaborative studies in food microbiology; Peterz M et al.; A method is presented for preparing samples of freeze-dried mixtures of microorganisms for proficiency tests and collaborative studies . The samples may include most microorganisms that are found in routine analysis in food laboratories . Transport of samples during 48 h did not decrease the number of microorganisms, nor was the variability among samples significantly affected by transport . The standard deviation of counts after 5 weeks of storage varied from 0.04 (Staphylococcus aureus) to 0.17 (Clostridium perfringens) log unit . Storage of samples for 10 weeks decreased the number of viable organisms by 0.02-0.43 log unit . Variability among samples increased for Providencia alcalifaciens and Bacillus cereus after 10 weeks of storage . No significant increase was found for the other organisms. Infect Immun, 1983 Nov, 42(2), 480 - 6 Intestinal colonization of infant hamsters with Clostridium difficile; Rolfe RD et al.; Infant hamsters of different ages were examined for their susceptibility to enteric Clostridium difficile colonization . Intragastric administration of C . difficile to infant hamsters resulted in multiplication of the organism in the intestinal tracts of animals 4 to 12 days old; hamsters younger or older were resistant to C . difficile intestinal colonization . Toxicity to the colonized animals could not be demonstrated despite cytotoxin titers in some infant hamsters comparable to titers found in the intestinal tracts of adult hamsters with C . difficile-associated intestinal disease . When introduced into 4-day-old hamsters, C . difficile colonized the intestinal tract and remained at high levels until the animals were 13 days old, at which time the presence of intestinal C . difficile could no longer be demonstrated . The number of C . difficile required to colonize the intestinal tracts of 50% of 7-day-old hamsters was 18 viable cells . On the other hand, 10(8) viable cells of C . difficile failed to colonize the intestinal tracts of healthy, non-antibiotic-treated adult hamsters. J Clin Pathol, 1983 Nov, 36(11), 1233 - 6 Use of gas-liquid chromatography as a screening test for toxigenic Clostridium difficile in diarrhoeal stools; Pepersack F et al.; In order to determine if gas-liquid chromatography (GLC) on concentrated stool extracts could be substituted to cell culture assay for cytotoxicity, we prospectively studied 154 diarrhoeal stools submitted for detection of Clostridium difficile toxin . Isocaproic-positive samples were cultured on egg yolk agar supplemented with cycloserine, cefoxitin and fructose for isolation of C difficile, and on egg yolk agar plus kanamycin for isolation of other clostridium species . Of the 154 samples, 129 were GLC-negative (height of the isocaproic peak less than 1.2 cm) and were toxin-negative . Twenty-five stools yielded isocaproic acid; C difficile isolated from 13 of them, six of which were also toxin-positive . Four other isocaproic-positive samples yielded C bifermentans and C sordellii; all were toxin-negative . These results indicate that a negative GLC is an excellent screening test for excluding C difficile infection; positive results must be checked by toxin testing and culture since they are not necessarily associated with the presence of C difficile or its toxin. Dis Colon Rectum, 1983 Nov, 26(11), 703 - 4 Nontraumatic Clostridium septicum gangrenous myonecrosis; Collier PE et al.; Nontraumatic Clostridium septicum infections may present as either septicemia or as metastatic myonecrosis . Most of these infections occur in debilitated patients with diabetes who are receiving cancer chemotherapy . The majority have a hematologic abnormality or a carcinoma of the colon . Usually there is an ulcerative lesion of the gastrointestinal tract that serves as the portal of entry . While most of these patients die from overwhelming sepsis, our patient was debrided early and treated promptly with high-dose penicillin therapy as well as hyperbaric oxygen therapy . He fully recovered from C . septicum gas gangrene and underwent resection of a recurrent colonic cancer. J Dairy Res, 1983 Nov, 50(4), 449 - 57 Antagonistic activity of Debaryomyces hansenii towards Clostridium tyrobutyricum and Cl . butyricum; Fatichenti F et al.; A study of the inhibitory action of Debaryomyces hansenii (31 strains) on Clostridium tyrobutyricum (5 strains) and Cl . butyricum (2 strains) on laboratory media showed that Deb . hansenii inhibited the growth of these organisms, and that this effect was due not only to competition for nutrients but also to the production of both extra- and intracellular antimicrobial metabolites . The inhibitory effect varied with strain and occurred whether the yeasts were grown aerobically or under reduced O2 tension. Coll Relat Res, 1983 Nov, 3(6), 445 - 58 Characterization of a large fragment from annelid cuticle collagen and its relationship to the intact molecule; Murray LW et al.; Digestion of the cuticle collagen from the annelid Nereis virens with Clostridium histolyticum collagenase yields a native, collagenase-resistant fragment (CCRF) of the molecule with an Mr of about 900,000 (Kimura and Tanzer, J . Biol . Chem . 252: 8018, 1977) . We have produced 940 nm long, SLS-crystallites from the collagenase-resistant fragment; the SLS pattern matches a region at one end of the 2,400 nm SLS obtained from intact cuticle collagen . Upon denaturation, the fragment yields two subunits, CCRFA and CCRFB, which can be separated by SDS polyacrylamide gel electrophoresis or by chromatography on DEAE-cellulose; the subunits are in about a 2:1 ratio . The subunits have amino acid compositions which are similar to those of the original A and B chains, although the fragments have a higher content of acidic residues and a lower content of hydroxyl residues . Previous studies of the intact B chain have shown that there is about one methionine in the chain, and that it is located near the COOH terminus . The CCRFB subunit also contains about one methionine, indicating that CCRFB is probably derived from the COOH end of the intact molecule . Based on these composite data, we have provisionally defined the amino and carboxyl ends of the cuticle collagen SLS and provide additional evidence that the molecule is a heteropolymer with the formula A(B)2. Appl Environ Microbiol, 1983 Nov, 46(5), 1169 - 75 Influence of carbohydrates on growth and sporulation of Clostridium perfringens in a defined medium with or without guanosine; Sacks LE; Clostridium perfringens strains NCTC 8238, NCTC 8798, NCTC 8679, 8-6, FD-1, and PS52 formed high levels of heat-resistant spores in a defined medium (D) with various sugars as energy sources . Strain PS49 formed high levels of heat-resistant spores when grown with dextrin and methylxanthines . The experiments showed the possibility of carrying out experiments on the sporulation of certain C . perfringens strains in a completely defined medium, without using the ill-defined polysaccharide dextrin . The addition of guanosine and sucrose to D medium generally suppressed sporulation in most strains and made it possible to prepare overnight cultures consisting mainly of vegetative cells . These cultures could be used to inoculate D medium directly, eliminating both the need to wash cells and the lag which normally occurs when cells have been grown in a different medium . Except for strains PS52 and NCTC 8238, guanosine generally increased growth rates and reduced sporulation for all strains when grown on simple sugars . Methylxanthines decreased growth rates and increased sporulation of NCTC 8679 and PS49 when present in D medium with dextrin . In the absence of guanosine, strains NCTC 8798 and 8-6 grew much slower on glucose than on disaccharides . Strain PS52 grew on lactose only after a prolonged lag . For strains requiring dextrin for good sporulation, a commercial dextrin (Difco Laboratories) was found to be readily filter sterilized, making it possible to prepare large amounts of media for use in the production of spores (or enterotoxin). Exp Cell Res, 1983 Nov, 149(1), 227 - 36 Enzymatic isolation of cells from neonatal calvaria using two purified enzymes from Clostridium histolyticum; Hefley TJ et al.; The enzymatic isolation of cells with bacterial collagenase has proved to be a powerful technique for the study of a wide variety of tissues . Unfortunately, for some applications such as the isolation of cells from membranous bone, the cellular damage that results from the exposure of the cells to cytotoxic contaminants of bacterial collagenase has limited the usefulness of this approach . The use of chromatographically purified collagenase alone is often ineffective or very slow to release cells from tissue . We have found that two enzymes are necessary and sufficient to isolate cells from neonatal mouse calvaria: purified collagenase and neutral protease . These two enzymes can be chromatographically purified on a preparative scale to yield 100 mg amounts of each enzyme . The purified enzymes can be recombined in amounts which will digest calvaria at the same rate as the crude bacterial collagenase from which they were derived . The cells that are isolated using the purified enzymes are undamaged, as indicated by the measurement of their equilibrium density on gradients of Ficoll and sodium metrizoate . Cells isolated with crude collagenase never reach an equilibrium density upon isopyknic centrifugation, whereas cells isolated with the purified enzymes reach an equilibrium density of 1.074 g/ml in 90 min. Mikrobiologiia, 1983 Nov-Dec, 52(6), 869 - 74 {Glucose metabolism in Clostridium sporogenes and Clostridium sticklandii bacteria}; Golovchenko NP et al.; Clostridium sporogenes 272 has a high rate of glucose fermentation . Its cell-free extract contains all glycolytic enzymes catalysing glucose degradation to pyruvate and shows the phosphoroclastic activity . C . sticklandii CSG has a low rate of glucose fermentation . Hence, the activity of the following enzymes is lower in this organism comparing to C . sporogenes: phosphohexoisomerase (EC 5.3.1.9), phosphofructokinase (EC 2.7.1.11), aldolase (EC 4.1.2.13), triosephosphate isomerase (EC 5.3.1.1) and glyceraldehyde phosphate dehydrogenase (EC 1.2.1.12) . Moreover, it is possible that the system of glucose transport into the cell is damaged in C . sticklandii. Proc Natl Acad Sci U S A, 1983 Nov, 80(22), 6799 - 803 Immunological crossreactivity of eukaryotic C1-tetrahydrofolate synthase and prokaryotic 10-formyltetrahydrofolate synthetase; Staben C et al.; Antiserum to yeast C1-tetrahydrofolate (C1-H4folate) synthase reacts with other eukaryotic C1-H4folate synthases and prokaryotic 10-formyltetrahydrofolate (10-CHO-H4folate) synthetases {formate:tetrahydrofolate ligase (ADP-forming), EC 6.3.4.3} even though these enzymes vary in subunit size and function and probably vary widely in sequence . The comigration of the purified enzymes with the immunoreactive material establishes the specificity of the reaction for C1-H4folate synthase proteins . Reciprocal crossreaction of the antibody to Clostridium acidiurici 10-CHO-H4folate synthetase with the eukaryotic proteins indicates that such broad cross-species reactions are not specific to the antisera elicited in response to the yeast C1-H4folate synthase . These specific crossreactions among divergent species have been observed only on an electrophoretic transfer blot of a denaturing polyacrylamide gel . These observations may have been possible because of the sensitivity and specificity of the technique, which differ from more conventional immunochemical methods. J Bacteriol, 1983 Nov, 156(2), 828 - 36 Characterization of a cellulose-binding, cellulase-containing complex in Clostridium thermocellum; Lamed R et al.; The isolation and biochemical characterization of the extracellular form of a cellulose-binding factor (CBF) from Clostridium thermocellum is described . The CBF was isolated from the culture supernatant by a two-step procedure which included affinity chromatography on cellulose and gel filtration on Sepharose 4B . The isolated CBF was homogeneous as determined by immunoelectrophoresis, polyacrylamide gel electrophoresis, gel filtration, and analytical ultracentrifugation analysis . The CBF was found to form a complex which exhibited a molecular weight estimated at 2.1 million . Electron microscopic analysis of negatively stained preparations of the isolated CBF revealed a particulate, multisubunit entity of complicated quaternary structure . The molecule appeared to be about 18 nm in size . Although urea failed to break the complex into its component parts, polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate resolved the CBF complex into 14 polypeptide bands . Immunoprecipitation experiments confirmed that these polypeptides indeed formed part of the same complex . Interestingly, by using the whole-cell immunization procedure described in the accompanying article (Bayer et al., J . Bacteriol., 156:818-827, 1983) only one CBF subunit (Mr = 210,000) was found to be antigenically active . By using a gel-overlay assay technique, at least eight of the remaining CBF-associated polypeptide components were shown to exhibit cellulolytic activity . The results are consistent with the contention that the CBF comprises a discrete, multisubunit complex or group of closely related complexes which exhibit separate antigenic and multiple cellulase activities in addition to the property of cellulose binding . It appears that the CBF is not only responsible for the adherence of the cells to cellulose but also constitutes a major part of the cellulolytic apparatus of this organism. J Biol Chem, 1983 Oct 25, 258(20), 12427 - 33 Hydrogenosomal ferredoxin of the anaerobic protozoon, Tritrichomonas foetus; Marczak R et al.; A low molecular weight iron-sulfur protein has been purified from Tritrichomonas foetus by deoxycholate extraction of whole cells, ion exchange chromatography, and gel filtration . The purified protein was essentially homogeneous as judged by isoelectric focusing, polyacrylamide gel electrophoresis, and gel filtration . A pI of 4.3 was observed . The molecular weight of the protein was estimated to be 12,000 . Chemical and spectral analysis showed the protein to have a {2Fe-2S} cluster . The absorbance spectrum of the oxidized protein showed maxima at 280, 340, 458 and shoulders at 410 and 550 nm . The maximum observed A458/A280 ratio was 0.82 and the absorbance of the oxidized protein at 458 nm was 8,000 M-1 X cm-1 . The low temperature EPR spectrum of the protein reduced with dithionite revealed axial symmetry with features at g values of g = 1.94 and g = 2.02 . The oxidized protein gave no EPR signal in the g = 1.8 to 2.2 range . Cell fractionation studies indicated the localization of this protein in the hydrogenosome . The protein was able to function as an electron transport component in the reduction of metronidazole (a 5-nitroimidazole derivative) by pyruvate:ferredoxin oxidoreductase and hydrogenase from T . foetus and also from Trichomonas vaginalis and Clostridium pasteurianum as well as in the reduction of cytochrome c by plant NADPH:ferredoxin oxidoreductase . This protein has the characteristics of a ferredoxin and is likely to be a physiological electron carrier in hydrogenosomal pyruvate oxidation. Biochim Biophys Acta, 1983 Oct 18, 760(2), 221 - 9 Characterization of the metalloproteinase inhibitor produced by bovine articular chondrocyte cultures; Morales TI et al.; Primary cultures of bovine articular chondrocytes release a latent metalloproteinase which is activated by incubation with organomercurials to degrade proteoglycans . All the enzyme present in the culture medium is latent and binds to columns of heparin-Sepharose . The yield of activity from the heparin-Sepharose columns (measured after organomercurial treatment) is approximately 300-1000% depending on the chondrocyte culture batch . Recombination of column fractions shows that the increase in activity is due to the separation of an inhibitor of the metalloproteinase by the chromatographic step . The metalloproteinase inhibitor has a molecular weight of approximately 35000 (determined by Bio-Gel P-60 chromatography) and binds reversibly to columns of concavalin A-Sepharose . It is relatively heat stable (30 min at 60 degrees C) and resistant to inactivation by trypsin (2 h, 37 degrees C, 10 microgram/ml trypsin) . The inhibitor is active against rat uterine collagenase and gelatinase but does not affect bacterial metalloproteinases such as thermolysin and Clostridium histolyticum collagenase. Exp Cell Res, 1983 Oct 15, 148(2), 413 - 22 Ultrastructural effects of Clostridium difficile toxin B on smooth muscle cells and fibroblasts; Wedel N et al.; The mechanism by which Clostridium difficile toxin B causes cells in culture to round was investigated . Cultured human lung fibroblasts and rabbit aortic smooth muscle cells were treated with partially purified or purified toxin B and monitored by light and transmission electron microscopy (TEM) . Both preparations caused progressive cell rounding which correlated with disorganization of actin-containing myofilament bundles . Thin myofilaments became fragmented and finally disappeared (after 24 h) and dense bodies became more prominent, while all other organelles appeared unaffected. Biochim Biophys Acta, 1983 Oct 12, 734(2), 257 - 66 A study on the topological distribution of phospholipids in microsomal membranes of chick brain using phospholipase C and trinitrobenzenesulfonic acid; Dominski J et al.; The transbilayer distribution of phospholipids in chicken brain microsomal membranes has been investigated using trinitrobenzenesulfonic acid and phospholipase C from Clostridium welchii . The exposure of intact microsomes to trinitrobenzenesulfonic acid showed that the labelling of aminophospholipids followed biphasic kinetics, indicating that these membranes contain a fast- and a slow-reacting pool of aminophospholipids . Use of microsomes radioiodinated on their surface led to the conclusion that the fast-reacting pool may be located on the outer leaflet of the microsomal vesicles . It contains about 35% of the phosphatidylethanolamine, 29% of the ethanolamine plasmalogens and 18% of the phosphatidylserine . The treatment of intact microsomes with the phospholipase C Cl . welchii produced the hydrolysis of 50% of the phospholipids without any loss of their permeability properties, indicating that they are not permeable to the hydrolase . Phospholipids extracted from the microsomes were hydrolyzed rapidly by the phospholipase C with the exception of phosphatidylserine and phosphatidylinositol . In intact microsomes about 90% of phosphatidylcholine, 32% of ethanolamine phospholipids and 60% of sphingomyelin were accessible to the phospholipase . These results suggest that the phospholipids have an asymmetric distribution in chicken brain microsomes, the external leaflet containing about 75% of the choline phospholipids and 25% of the aminophospholipids, whereas an opposite distribution is observed in the inner leaflet. J Gen Microbiol, 1983 Oct, 129 (Pt 10), 3227 - 37 Partial purification and characterization of two enzymes involved in isovaleric acid synthesis in Clostridium bifermentans; Britz ML et al.; Conversion of leucine to isovaleric acid by Clostridium bifermentans is achieved by the action of at least two enzymes . One is a transaminase producing alpha-ketoisocaproic acid, which was purified 30-fold from osmotic lysates of late-exponential phase cells by repeated chromatography on DEAE-Sepharose C16B and Sephacryl S300: this represented a 147-fold purification of activity found in sonically disrupted cells . This enzyme had an apparent molecular weight of approximately 190000 and was composed of six identically sized sub-units (molecular weight 31000 +/- 1000) . Transamination required pyridoxal phosphate and pyruvate and was optimal at pH 8.6; the apparent Km for leucine was 7.0 mM . Activity was totally inhibited by 1 mM-p-chloromercuribenzoate and partially inhibited by other thiol reagents . The second enzyme decarboxylated alpha-ketoisocaproic acid to form isovaleric acid and was also partially purified by chromatography on DEAE-Sepharose C16B and Sephacryl S300 . It has an apparent molecular weight of 240000 and required FAD and coenzyme A for activity; the Km for alpha-ketoisocaproic acid was 4.2 mM and activity was optimal around pH 8.0 . This enzyme was a flavoprotein with absorption maxima at 280, 320 and 400 nm, and a fluorescent maximum at 500 nm . The prosthetic group, FAD, dissociated from the protein during purification resulting in an inactive apoenzyme which was only partially re-activated by FAD . Activity was completely inhibited by several thiol reagents tested at 1 mM. J Steroid Biochem, 1983 Oct, 19(4), 1441 - 50 Tetra- and hexahydro derivatives of aldosterone and 18-hydroxycorticosterone by chemical and microbial reductions; Harnik M et al.; Preparative methods were developed for reduction with NaBH4 at 0 of 3 beta, 5 alpha- and 3 alpha, 5 beta-tetrahydroaldosterone (1) and (12) to their respective 20 alpha-ol derivatives 2a and 13a . Corroboration of structures was obtained by periodate oxidations to the lactols 3b and 14b and thence, by further oxidation, to the lactones 4 and 15 respectively; these lactones were also independently obtained from 1 and 12 . Reduction with NaBH4 at 80 degrees C converted 1 and 12 into 18-hydroxy-3 beta, 5 alpha, 20- and 18-hydroxy-3 alpha, 5 beta, 20-hexahydrocorticosterone 6a and 17a respectively, which were mixtures of epimers at C-20 . Compound 17a could also be prepared by reduction of the lactone 21 with sodium aluminum bis-(methoxyethoxy) hydride . Again, periodate oxidations of 6a and 17a gave the lactols 7b and 22b and thence, by Jones oxidation, the diketolactones 8 and 23, which were also prepared from 18-hydroxy-11-dehydrocorticosterone (10) and 18-hydroxycorticosterone (24) respectively . Improved conditions for reduction with Clostridium paraputrificum permitted convenient conversion of aldosterone (11), the corresponding 18 leads to 11 lactone 18a and 18-hydroxycorticosterone (24) into their 3 alpha, 5 beta-tetrahydro derivatives. J Comp Pathol, 1983 Oct, 93(4), 597 - 601 Clostridium fallax as a cause of gas-oedema disease in a horse; Coloe PJ et al.; We record a fatal case of gas-oedema disease (malignant oedema) in a 5-year-old horse . The nature of the lesion is consistent with a gas-oedema type infection due to a Clostridium spp . The causative organism was isolated and identified by conventional biochemical tests and by gas-liquid chromatographic analysis of its metabolic products as Clostridium fallax, but significant variations in the reported biochemical characteristics of Cl . fallax were detected . We believe that this is the first reported case of Cl . fallax infection in a horse. J Antimicrob Chemother, 1983 Oct, 12(4), 347 - 56 Susceptibility of clostridia from farm animals to 21 antimicrobial agents including some used for growth promotion; Dutta GN et al.; The minimal inhibitory concentrations of 21 antimicrobial agents were determined by an agar dilution method against 68 strains of 18 Clostridium species isolated from caeca of pigs, cattle and poultry . Of the therapeutically used antibiotics, chloramphenicol was most active in vitro followed by penicillin G, the lincosamides and tetracycline . Penicillin-resistant Cl . butyricum strains produced a beta-lactamase . Avoparcin, carbadox, monensin, nitrovin and virginiamycin were the most effective of the growth promoting antimicrobial agents . Many clostridial species were naturally resistant to flavomycin and Cl . sporogenes was naturally resistant additionally to bacitracin, tiamulin, the lincosamides and virginiamycin component M . The macrolide, lincosamide and streptogramin resistant strains showed four different patterns of resistance to the antibiotics . One of these resistance patterns was the natural resistance of Cl . sporogenes to the lincosamides and virginiamycin component M which is a streptogramin group A antibiotic. J Clin Microbiol, 1983 Oct, 18(4), 1017 - 9 Efficiency of various bile salt preparations for stimulation of Clostridium difficile spore germination; Wilson KH; Taurocholate, desoxycholate, and cholate stimulated germination of Clostridium difficile spores in broth medium and enhanced recovery of C . difficile spores on a selective agar medium . Desoxycholate and some crude taurocholate preparations also inhibited multiplication of vegetative cells . At a concentration of 1.2 X 10(-2) M, sodium cholate inhibited multiplication of vegetative cells, but at concentrations of 1.2 X 10(-3) to 2.4 X 10(-3) M, it stimulated germination without inhibiting cell multiplication . Thus, pure sodium taurocholate and sodium cholate may effectively be incorporated in cefoxitin-cycloserine-fructose agar, whereas some crude preparations of sodium taurocholate decrease recovery on this medium. J Clin Microbiol, 1983 Oct, 18(4), 1006 - 7 Concomitance of cytotoxigenic and non-cytotoxigenic Clostridium difficile in stool specimens; Borriello SP et al.; Six patients with antibiotic-associated diarrhea and one patient with diarrhea unrelated to antibiotic use yielded both cytotoxigenic and non-cytotoxigenic isolates of Clostridium difficile from the same stool specimens . In addition, these isolates were shown to be pathogenic and nonpathogenic, respectively, in the hamster model of antibiotic-associated colitis . These data imply that more than one toxin type of C . difficile may be harbored simultaneously . If toxin testing is used to identify C . difficile, more than one colony must be tested. J Clin Pathol, 1983 Oct, 36(10), 1184 - 7 Pseudomembranous colitis associated with changes in an ileal conduit; Shortland JR et al.; A case of antibiotic associated pseudomembranous colitis following total cystectomy is reported, in which there was involvement of the ileal conduit . The small bowel remaining in situ was uninvolved . Bacteriological studies revealed Clostridium difficile and the toxin in both colon and ileal conduit . Relevant publications concerning pathogenesis are discussed, in relation to the unusual site described in this case . Epidemiological evidence is reviewed which suggests that isolation of patients with pseudomembranous colitis is a logical course of action. J Wildl Dis, 1983 Oct, 19(4), 302 - 7 Prevalence of Clostridium botulinum type C in substrates of phosphate-mine settling ponds and implications for epizootics of avian botulism; Marion WR et al.; Prevalence and conditions for occurrence of Clostridium botulinum type C were examined on phosphate-mine settling ponds and a natural wetland in northern Florida between April 1981 and March 1982 . Substrate samples were collected monthly (winter) and semi-monthly (summer) from 16 locations on seven ponds . Selected environmental parameters were measured at each location at the time of sampling . Mouse inoculation tests and toxin neutralization tests using enrichment culture filtrates were conducted to identify C . botulinum type C in the samples . The bacteria were identified in 26 (5.6%) of 467 sediment samples . Occurrences were distributed over four of the seven ponds and included nine of the 16 sample locations, but were restricted to the months April through October . The organism occurred over a wide range of ecological conditions found on the ponds during these months . If the presence of C . botulinum type C in the substrate is a prerequisite for botulism to occur, the prevalence and fairly wide distribution of this organism on settling ponds makes it difficult to predict where future outbreaks may occur. Appl Environ Microbiol, 1983 Oct, 46(4), 961 - 3 Interaction of pH and NaCl on culture density of Clostridium botulinum 62A; Montville TJ; Clostridium botulinum 62A growth rates declined with decreasing pH and increasing salt levels . Lysis rates, however, were affected only by pH . Due to competition between growth and lysis rates, an accurate assessment of interactive effects was obtained only when optical density determinations were made at multiple intervals. Infect Immun, 1983 Oct, 42(1), 64 - 70 Proteolysis of sialoglycoprotein by Pasteurella haemolytica cytotoxic culture supernatant; Otulakowski GL et al.; Proteolytic enzyme activity releasing sialo glycopeptides from 3H-labeled human erythrocyte ghosts was detected in cytotoxic (leukotoxic) culture supernatants from 9 of 12 Pasteurella haemolytica serotypes . Microcrystalline cellulose thin-layer chromatograms of radioactive water-soluble products showed the following two radioactive peaks: a high-mobility minor peak (Rf, 0.54 to 0.74), identified as sialic acid, and a low-mobility major peak (Rf, 0.18 to 0.21), partially characterized as a trichloroacetic acid-soluble, sialic acid-rich fragment with a molecular weight of greater than 3,500, not extractable by chloroform . The sialic acid content of this fragment after treatment with Clostridium perfringens neuraminidase was estimated to be 7.2 X 10(-2) mumol mg-1 . The presence of neuraminidase as a separate activity in some culture supernatants was confirmed . It is considered to be responsible for the observed release of free sialic acid . Preliminary studies with the crude enzyme showed that it has a broad pH optimum around pH 7.0 and that activity is not affected by inhibitors of trypsin, chymotrypsin, thermolysin, thio and serine enzymes, nor by an inhibitor of neuraminidase, 2,3-dehydro-2-deoxy-N-acetylneuraminic acid . Activity was, however, inhibited by o-phenanthroline at a high concentration after prolonged treatment . The enzyme hydrolyzed glycophorin at a rate four times higher than the rate for casein . Free glycophorin inhibited the enzyme-induced release of radioactive products from 3H-labeled ghosts . It is speculated that the novel enzyme is a neutral protease, probably metal-dependent, with specificity for sialoglycopeptides . The possible relationship of this protease to the previously reported host species-specific leukotoxicity of P . haemolytica and its potential role in virulence is discussed. J Nutr Sci Vitaminol (Tokyo), 1983 Oct, 29(5), 601 - 9 Influence of diets low in protein or lysine on the cecal flora of rats with reference to cecal contents; Takahashi M et al.; To determine the effect of a certain diet on the intestinal flora of rats, the cecal flora of rats fed a low protein or low lysine diet was examined . Total counts of bacteria in the cecal contents of rats given the four kinds of diet (a normal protein diet, a low protein diet, a normal lysine diet and a low lysine diet) were not significantly different . The counts of Streptococcus, Enterobacteriaceae, and Clostridium perfringens in the cecal contents from rats fed the low protein diet were significantly lower than those from rats fed the normal protein diet . The count of Lactobacillus in the cecal contents of the low protein group were significantly lower than those of the control . But no significant difference was found between the levels of most of the free amino acids in the cecal contents of the low lysine group and those of the control group. Can J Microbiol, 1983 Oct, 29(10), 1470 - 4 A new inorganic pyrophosphate utilizing bacterium from a stagnant lake; Varma AK et al.; An apparently new species of Clostridium was isolated from mud samples obtained from a stagnant lake near Athens, Georgia . These organisms were able to utilize inorganic pyrophosphate as a source of energy for growth, a phenomenon representing the simplest ATP-generating system in the biological world. Can J Microbiol, 1983 Oct, 29(10), 1241 - 6 Transferable tetracycline resistance in Clostridium perfringens strains of porcine origin; Rood JI; These studies represent the first systematic survey of the incidence of conjugative antibiotic resistance in Clostridium perfringens . Ninety-two antibiotic-resistant porcine strains were examined to see if they could donate their antibiotic-resistance determinants to sensitive recipient strains . Fifteen of the 89 tetracycline-resistant strains transferred their tetracycline resistance in mixed-plate mating experiments but no transfer of macrolide-lincosamide resistance was detected . The efficiencies of transfer of tetracycline resistance varied from 1.3 X 10(-3) to 1.9 X 10(-6) transconjugants per donor cell . Significantly higher transfer efficiencies were observed when both the donor and recipient strains were derivatives of strain CW 362 . These values ranged from 3.7 X 10(-1) to 4.6 X 10(-2) transconjugants per donor cell . This high frequency transfer system should prove invaluable for further genetic studies on this microorganism. J Biochem (Tokyo), 1983 Oct, 94(4), 1053 - 9 Purification and some properties of nitrite reductase from Clostridium perfringens; Sekiguchi S et al.; Nitrite reductase from Clostridium perfringens was purified by chromatographies on DEAE-cellulose, DEAE-Sephadex, Sephadex G-150, and hydroxylapatite and by isoelectric focussing to a homogeneous state, showing essentially a single protein band in disc gel electrophoresis and a single immuno-precipitation line in double diffusion against antiserum obtained from immunized rabbits . The reductase was induced in the presence of nitrate . It had a molecular weight of 54,000 and showed no absorption peak in the visible region . The pH optimum was 6.2 and Km for nitrite was 5 mM . Ferredoxin, as well as viologen dyes, was found to be an electron donor . The product of nitrite reduction was hydroxylamine . This reductase was inhibited by o-phenanthroline and azide but not by cyanide or diethyldithiocarbamate. J Appl Bacteriol, 1983 Oct, 55(2), 203 - 8 Egg-yolk trypticase soy agar for the enumeration of heat-damaged spores of Clostridium sporogenes; Michels MJ et al.; In heat-resistance studies with spores of Clostridium sporogenes BC-2, an improved recovery medium was needed for severely heat-damaged spores as the used previously--Wynne medium in Miller-Prickett tubes--did not allow accurate counts of spores because of gas formation and disruption of agar . Initial test with pour plates of Viande-Leyure medium containing egg-yolk gave much increased counts for spores previously heated for 50 min at 112 degrees C; this increase was attributed to the presence of egg-yolk . Addition of egg-yolk to Reinforced Clostridial Agar, All-Culture Medium and Trypticase Soy Agar showed that Trypticase Soy Agar with egg-yolk was the best recovery medium . For the final formulation, the value of supplementation with cysteine-HCl and methylene blue was also shown . The resultant Egg-yolk Trypticase Soy Agar is conveniently prepared from BBL Trypticase Soy Agar (40 g) with the addition of 0.4 g/l cysteine-HCl, 4 mg/l of methylene blue and 2% Oxoid egg-yolk emulsion aseptically to the melted basal medium . For optimal spore counts, pour plates are incubated anaerobically for 5--7 d at 30 degrees C. Biochem Biophys Res Commun, 1983 Sep 15, 115(2), 658 - 65 13C and 61Ni isotope substitutions confirm the presence of a nickel (III)-carbon species in acetogenic CO dehydrogenases; Ragsdale SW et al.; The nickel-containing CO dehydrogenases from Acetobacterium woodii and Clostridium thermoaceticum were studied by EPR spectroscopy in order to define the components involved in the EPR spectrum obtained by reaction of the enzymes with the substrate, CO . Using isotopic substitution techniques, these experiments unequivocally establish that a nickel-carbon species is involved in the g = 2.08, 2.02 EPR signal . Comparing the 61Ni- and 59Ni-substituted enzymes, the g = 2.08 component of the resonance was found to be mainly due to nickel with a smaller contribution by the carbon species . Reaction of the CO dehydrogenase with {13C}CO versus {12C}CO showed that a carbon species, formed from CO, was the major contributor to the g = 2.02 EPR signal . In addition, the oxidized CO dehydrogenase was found to exhibit a Ni (III) EPR signal analogous to that of the hydrogenases from the methanogenic and sulfate-reducing bacteria. Biochemistry, 1983 Sep 13, 22(19), 4472 - 80 Influence of pN2 and pD2 on HD formation by various nitrogenases; Li JL et al.; Formation of HD from D2 has been demonstrated with nitrogenase preparations from Azotobacter vinelandii, Clostridium pasteurianum, Klebsiella pneumoniae, and Azospirillum sp . We conclude that the formation of HD from D2 is a general property of nitrogenases . However, the nitrogenases differ in their Ki values for D2 (N2 fixation) and in their rates of catalyzing HD formation; among the nitrogenases tested, C . pasteurianum nitrogenase had the lowest activity for formation of HD . When contaminating N2 was removed from the atmospheres above reaction mixtures, less than 1% of the total electron flux in the system was directed to HD formation; hence, we doubt that N2-independent HD formation is significant . A working hypothesis is suggested that operates without invoking an N2-independent reaction for forming HD. Biochemistry, 1983 Sep 13, 22(19), 4556 - 61 Inhibition of collagenase from Clostridium histolyticum by phosphoric and phosphonic amides; Galardy RE et al.; Di- and tripeptides with sequences present in collagen that are known to occupy the S1' through S3' subsites at the active site of the collagenase from Clostridium histolyticum do not themselves inhibit this zinc protease . Thus glycylproline, glycylprolylalanine, and their C-terminal amides are not inhibitors . N alpha-Phosphorylglycylproline, N alpha-phosphorylglycyl-L-prolyl-L-alanine, and their C-terminal amides are weak inhibitors with IC50's (concentration causing half-maximal inhibition) of 4.6, 0.8, 3, and 1.5 mM, respectively . Extension of glycyl-L-prolyl-L-alanine to L-leucyl-glycyl-L-prolyl-L-alanine gives a tetrapeptide known to occupy the S1, S1', S2', and S3' subsites of collagenase when present in collagen but that still does not itself inhibit the enzyme . (Isoamylphosphonyl)glycyl-L-prolyl-L-alanine, a peptide containing a tetrahedral phosphorus atom at the position of the amide carbonyl carbon of the L-leucylglycyl amide bond of the parent tetrapeptide, inhibits collagenase with an IC50 of 16 microM, at least 1000-fold more potent than the parent peptide . Substitution of the two-carbon ethyl chain of alanine for the five-carbon isoamyl chain of leucine increases the IC50 to 46 microM . Substitution of the n-decyl chain for the isoamyl chain does not change the IC50 . (Isoamylphosphonyl)glycyl-glycyl-L-proline contains a tripeptide that does not occupy the S1' through S3' subsites of collagenase when this peptide is present in collagen and thus has an IC50 of 4.4 mM . (Isoamylphosphonyl)glycyl-L-prolyl-L-alanine may be an analogue of the tetrahedral transition state for the hydrolysis of the natural collagen substrate.(ABSTRACT TRUNCATED AT 250 WORDS) Burns Incl Therm Inj, 1983 Sep, 10(1), 17 - 29 1981 circus fire disaster in Bangalore, India: causes, management of burn patients and possible presentation; Das RA; The circus fire disaster claimed 92 lives and 300 others were injured . A total of 119 patients were treated in the Burns centre at Victoria Hospital . Forty-two patients were treated as outpatients and 77 cases were admitted . Fourteen patients with more than 80 per cent burns of the body surface died within 48 hours of the disaster . Three patients out of the remaining 63 cases died in the course of treatment, 32 patients were operated by escharectomy and skin grafting or flap procedures . Proper medical assessment, early fluid therapy and respiratory care saved many critical patients . Human Antitetanus toxin and Pseudomonas hyper Immune globulin seem to have a definite role in the prevention and control of infection with Clostridium tetani and Pseudomonas aeruginosa 'Furacin' was found to be a valuable topical agent. Pediatr Infect Dis, 1983 Sep-Oct, 2(5), 364 - 6 Lack of relationship of Clostridium difficile to antibiotic-associated diarrhea in children; Elstner CL et al.; We studied prospectively the conversion rate to Clostridium difficile-positive stool cultures in 31 children receiving oral antibiotics for common infections and looked for a possible association of C . difficile colonization with diarrhea . The incidence of pretreatment positive stool cultures was 35% with the majority of positive findings in infants less than 1 year of age . After treatment with oral antibiotics C . difficile was cultured from the stool of 42% of the children . Eleven children developed diarrhea during antibiotic therapy . Seven of these children had at least one stool culture positive for C . difficile and four had persistently negative cultures . Oral antibiotic treatment of common infections in otherwise healthy children does not appear to predispose to stool colonization with C . difficile, nor is the presence of C . difficile in stools in these children significantly associated with the onset of antibiotic-associated diarrhea. J Gen Microbiol, 1983 Sep, 129 (Pt 9), 2837 - 45 Identification of Clostridium butyricum and Clostridium beijerinckii by gas-liquid chromatography and sugar fermentation: correlation with DNA homologies and electrophoretic patterns; Magot M et al.; Sixty-five strains of clostridia of the butyricum group were studied by DNA-DNA hybridization, electrophoresis of cell proteins, gas-liquid chromatography, and fermentation of glycerol, inositol and ribose . The DNA--DNA hybridization results confirmed that strains of this group belong to two main species, Clostridium butyricum and C . beijerinckii . Five strains did not hybridize with the reference strains of these two species . Most of the strains could be identified by quantitative gas-liquid chromatographic analysis combined with fermentation patterns . The other strains could be identified by their protein electrophoretic patterns. J Clin Microbiol, 1983 Sep, 18(3), 733 - 4 Inoculum preparation for anaerobic susceptibility tests; Murray PR et al.; The results of anaerobic susceptibility tests performed with inocula prepared directly from agar isolation media and from overnight broth cultures were compared . Altogether, 93.0% of the results with these two inoculum preparations were within one twofold dilution, including 92.5 and 95.6% of the results with Clostridium species and the Bacteroides fragilis group, respectively . Thus, inocula prepared from agar subculture plates resulted in more-timely susceptibility test results . In addition, the test organisms used in this study grew better when the inoculum was prepared directly from agar plates. J Clin Microbiol, 1983 Sep, 18(3), 491 - 4 Isolation of Clostridium pseudotetanicum from a patient with gas gangrene; Katoh N et al.; Clostridium pseudotetanicum was isolated along with Serratia marcescens, Proteus mirabilis, and Staphylococcus epidermidis from a patient suffering from gas gangrene who had been injured in the right leg by a power cultivator . Experimental infection of the hind leg of mice with C . pseudotetanicum and the three kinds of aerobic bacteria did not produce any different macroscopic finding in the infection site, compared with aerobic bacterial injection, except for some enlargement of the involved tissue and some slightly altered histolytic findings . Ampicillin, rifampin, and tinidazole were the most active antimicrobial agents against C . pseudotetanicum. J Lipid Res, 1983 Sep, 24(9), 1119 - 26 Separation of 7 alpha- and 7 beta-hydroxysteroid dehydrogenase activities from clostridium absonum ATCC# 27555 and cellular response of this organism to bile acid inducers; Macdonald IA et al.; Both 7 alpha- and 7 beta-hydroxysteroid dehydrogenases (HSDH) were induced by either chenodeoxy-(CDC) or deoxycholic (DC) acid in C . absonum . 7 beta-HSDH was partially purified 35-fold from CDC-induced cultures of C . absonum by Procion Red (PR) affinity chromatography and high performance liquid chromatography (HPLC) using a TSK 3000 SW gel filtration column . A relative molecular weight of 200 K was estimated for 7 beta-HSDH using Sephacryl S-300 chromatography . Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the 35-fold purified 7 beta-HSDH showed six polypeptides in the molecular weight range of 40-50 K . Induction of cultures of C . absonum with CDC or DC (0.4 mM) also resulted in the differential synthesis of at least five new polypeptides with molecular weights of 94 K, 42 K, 32 K, 21 K, and 16 K . The 16 K polypeptide was induced by DC but not by CDC . SDS-PAGE of Triton X-100-solubilized membranes from these extracts revealed the presence of a new membrane-associated polypeptide of molecular weight 80 K . The soluble inducible polypeptides were eliminated during purification of the 7 alpha- and 7 beta-HSDH and, therefore, are not required for these enzyme activities . It is proposed that this organism synthesized 7 alpha- and 7 beta-HSDH as well as a series of other proteins in response to bile acids which may, in the absence of the dehydrogenases, be toxic to C . absonum . The HSDH's catalyze the epimerization of chenodeoxycholic acid to ursodeoxycholic acid, which is less toxic than the chenodeoxycholic acid . The other proteins may assist the survival of the organism in a high bile acid environment by mechanisms not yet understood. Biokhimiia, 1983 Sep, 48(9), 1548 - 54 {Molecular structure and immunochemical properties of highly purified hemagglutinin from Clostridium botulinum type A}; Ivanova LG et al.; A procedure for isolation of highly purified hemagglutinin from a toxic complex of culture filtrates of Cl . botulinum type A is described . This procedure includes precipitation with (NH4)2SO4, chromatography on Sephadex G-100, G-200 and DEAE-cellulose, specific adsorption on human erythrocytes and affinity chromatography . Using polyacrylamide gel electrophoresis, it was shown that hemagglutinin is a heteropolymeric protein consisting of a monomer (Mr 53 000) and a trimer (Mr 160 000) . The monomer is made up of two subunits with Mr 13 000 and one subunit with Mr 27 000 covalently linked by SS-crosslinks . The number and nature of the SS-crosslinks and SH-groups in the protein molecule were determined and a hypothetical structural model of hemagglutinin was proposed . Using immunochemical analysis, it was shown that some (but not all) serological properties of the highly purified protein from Cl . botulinum type A and of its partially purified counterpart are similar to those of hemagglutinin from Cl . botulinum type B. Public Health Rep, 1983 Sep-Oct, 98(5), 412 - 5 The Food and Drug Administration's role in the canned salmon recalls of 1982; Hayes AH Jr; The Alaska salmon industry conducted 9 recalls of 7 3/4-oz cans of salmon in 1982 after a 7 3/4-oz can of Alaskan salmon was implicated in illness and one death in Belgium from Clostridium botulinum type E toxin . By the code number on the can, the Food and Drug Administration (FDA), Seattle District, traced it to a specific salmon packer . Subsequently, the FDA received a report about a defect in the can . Investigation of the salmon packer's plant by the Agency revealed that the equipment used at the plant to reform the cans--which arrived at the cannery in a nearly flattened state--might have been responsible for the defect . The death and illness in Belgium, combined with the results of the FDA inspection of the plant implicated in the Belgian incident, provided strong evidence of the existence of a hazardous situation that might have widespread adverse health effects . The Food and Drug Administration therefore requested the firm to recall its 1980 and 1981 production of salmon packaged in 7 3/4-oz cans . The Agency then began an investigation of all U.S . salmon packed inn cans of this size that had been reformed on the equipment implicated in the can defect . Of 300,000 cans examined, 22 with the defect were found . As additional firms were identified as having used the defective cans, subsequent recalls were initiated. J Clin Microbiol, 1983 Sep, 18(3), 614 - 21 Comparison of three methods for anaerobe identification; Appelbaum PC et al.; In this study we evaluated the ability of three commercial methods, API 20A (Analytab Products, Plainview, N.Y.), Minitek (BBL Microbiolog