Clostridia

J Gen Microbiol, 1976 Jan, 92(1), 81 - 8
The effect of sublethal doses of rifampin on the sporulation of Clostridium botulinum; Hawirko RZ et al.; Sublethal doses of rifampin (0-005 mug/ml), added to vegetatively growing cultures of a sporogenic mutant of Clostridium botulinum at inoculation time or after 4 h, resulted in a decrease of growth and in blockage of spore formation . But when rifampin was added 6 to 24 h after inoculation, normal growth and sporulation occurred, indicating that the time of addition was critical and that rifampin was most effective on rapidly dividing, exponential-phase cells . Ultrastructural studies showed that when rifampin was added at the time of inoculation, endospore development was blocked at stage III . During subsequent incubation (greater than 10 h) the cells lost their rigidity, and lysis of the mother cell was followed by that of the forespore . When the cultures were treated with rifampin at 4 h, about 40% of the cells were blocked at stage III and about 60% reached stages IV and V . Some showed excessive elongation and contained developing spores at each pole . They appeared to be derived from two daughter cells unable to form a division septum because of a specific inhibitory effect of rifampin on division . It would seem, therefore, that two daughter cells which are genetically coded to form endospores will do so irrespective of the development of a division septum, and the spores are formed at the 'old' polar regions.

J Gen Microbiol, 1976 Jan, 92(1), 67 - 75
The stability of toxigenicity in Clostridium botulinum types C and D; Oguma K; Several type C and D strains of Clostridium botulinum, which had been converted to the toxgenic state by phages, were serially transferred through cooked meat medium with and without specific anti-phage serum . Most of the converted strains lost their toxigenicity even during transfer without antiserum, and the non-toxigenic variants that appeared were resistant to lysis and conversion by the original phage . However, in some combinations of phage and host bacteria toxigenicity was stable after ten transfers, though it showed a transient decrease, and the non-toxigenic variants that arose remained sensitive to lysis and conversion . When converted strains were transferred in medium containing anti-phage serum, toxigenicity was lost more rapidly than in the absence of serum and the non-toxigenic variants that appeared remained sensitive to lysis and conversion by the parent phage . Filtrates of the supernatants of culture fluids of strains transferred without anti-phase serum converted non-toxigenic strains to toxigenicity at varying rates.

Acta Chir Scand, 1976, 142(6), 461 - 6
Pathophysiology in the acute afferent loop syndrome . A study in rats; Bengtsson S et al.; The afferent loop syndrome, i.e . occlusion of an afferent intestinal loop after a Billroth II partial gastrectomy, was induced in rats . After various time intervals the animals were killed and the haematocrit and serum osmolality were determined . The content of the occluded loop was analysed with respect to volume, bacterial flora and osmolality . In some cases a sample of the content was incubated at 37 degrees C and the osmolality determined at regular intervals . Groups of animals were studied in this way after 30 min or 1, 4, 8 or 12 h of occlusion . The haematocrit rose with time after the occlusion . The osmolality of the plasma and of the content of the occluded loop did not increase . The volume of fluid in the occluded loop increased continuously with time--from an average of 0.5 ml in the control cases to 6.1 ml after 12 h of occlusion . Experiments in vitro showed that the initial osmolality of the content of the loop was 300 mOsm . After incubation of the samples this increased by 43 to 146 percent . The bacterial content of the loop, including Clostridium perfringens, increased significantly . The results indicate that a marked breakdown of substances takes place in such an occluded intestinal loop, which increases the osmolality . As a result fluid is immediately attracted to the loop to keep the osmolality constant . A combination of this fluid increase due to osmosis and contractions of the intestinal wall leads to a pressure in the occluded loop which considerably exceeds the pressure in the common bile duct due to secretions from the pancreas and liver.

Dev Biol Stand, 1976, 32, 151 - 5
{Failure of anti-symptomatic vaccination and the methods of vaccine activity control}; Joubert L et al.; Blackleg (Clostridium chauvoei) infection in Charollais cattle appears to have undergone some etiological and pathogenic changes which are reflected in the apparent failure of vaccination . Two methods have been used to determine the quality of the vaccines in order to meet the different requirements specified by pharmacopeias . The two methods differ in the vaccination schedules of the guinea pigs and the test strain used which may be either a virulent culture or a suspension of spore in calcium chloride . Both methods appear to be efficient in selecting vaccines which provide good protection . It follows, therefore, that the vaccination failures would appear to be the result of intensive selection of beef cattle with reduced immunological response . If this is the case, the remedy for these vaccination failures might lie in an adaptation of the vaccination schedule to the early developing breeds of cattle obtained by intensive selective breeding.

Dev Biol Stand, 1976, 32, 143 - 50
Some problems concerning the assay of Clostridium chauvoei vaccine in laboratory animals; Knight PA et al.; The potency test prescribed for Cl . chauvoei vaccines in the British Veterinary Codex requires the complete survival of a group of immunized guinea pigs and death of all unvaccinated controls . Problems arising from the influence of extraneous factors such as animal strain, diet and seasonal variation on the outcome of this test are discussed together with alternative methods that might permit the use of a standard preparation.

Dev Biol Stand, 1976, 32, 137 - 41
Some observations on the quality control testing of Clostridium chauvoei vaccines; Chandler HM; Agglutination tests are not suitable for the estimation of the protective antibody level in the sera of vaccinated animals and should not be used for the quality control testing of blackleg vaccines . Some highly virulent strains of Cl . chauvoei are only effectively protected against by vaccines containing cells with a heat labile protective antigen in addition to the heat stable antigen common to all cells of Cl . chauvoei . As these highly virulent strains may be encountered in the field it is important that such strains should be selected for the challenge of vaccinated animals in the quality control testing of blackleg vaccines . It would be useful if an international center could be responsible for the collection and distribution of such strains.

Dev Biol Stand, 1976, 32, 123 - 30
Preparation and evaluation to Clostridium septicum toxoid; Kennedy K; Clostridium septicum was grown under controlled conditions to produce high levels of alpha toxin . The toxin was toxoided and concentrated by ultrafiltration . The toxoided alpha toxin retained immunizing properties . The concentrated toxoid, when formulated on Total Combining Power, in monovalent or multivalent clostridial products, provided protection to rabbits against direct challenge with Cl . septicum spores . The toxoided alpha toxin produced antitoxin levels which exceeded current minima.

Dev Biol Stand, 1976, 32, 115 - 22
Potency testing of Clostridium septicum bacterins in sheep and laboratory animals; Cardella MA et al.; The immunogenic potency of bacterins containing Clostridium septicum was evaluated in mice, hamsters, guinea pigs, rabbits and sheep to establish potency requirements for these bacterins . Two subcutaneous injections of Cl . septicum bacterins were shown to provide resistance against intramuscular spore challenge . Protection in mice, hamsters and guinea pigs was generally at low levels, in the range of 1 to 5 LD50 . Efficacy estimates, at these levels of challenge, indicated that a degree of relationship existed between the response of rabbits and the response of sheep . This relationship was confirmed at higher levels of spore challenge, at greater than 5 to 500 LD50 . Undiluted bacterins were classed as good or poor based on percent survival responses in sheep . Bacterins providing 90 to 100% survival were classed as good; those providing 30 to 57% survival were classed as poor . In rabbits, this classification was less evident with the undiluted bacterins . Dilution of bacterins to 1:32 provided a discriminatory level with 7 of 8 bacterins examined and represented correlation of the sheep and rabbits responses . A similar relationship was demonstrated between protection in sheep against spore challenge and the antitoxin response in rabbits to undiluted bacterins . Antitoxin responses, when obtained, were generally higher in rabbits than those obtained in sheep . Further evaluation of a good and a poor bacterin in rabbits indicated that no direct relationship existed between antitoxin titer in rabbits and resistance to spore challenge, when compared at the same dilutions of bacterins . Evaluation of serum agglutination responses to types I and II somatic antigens indicated a poor relationship between agglutination response and resistance to spore challenge in both sheep and rabbits . Cl . septicum toxoids were shown to provide resistance to spore challenge in rabbits.

Dev Biol Stand, 1976, 32, 103 - 12
Clostridium oedematiens: observations on potency assaying; Macheak ME; The United States has established a Standard Requirement for the potency testing of biological products containing Clostridium oedematiens belonging to two types: type B (Cl . novyi) and type D (Cl . haemolyticum) . Guinea pig testing has provided widely varying results depending on the origin of the animals . The tests reported are intended to determine the efficacity of the vaccines (Cl . novyi and Cl . haemolyticum) depending on the animal tested: guinea pigs, sheep and bovines; they further establish a parallelism between the antitoxin titer and the immunity of the animal.

Klin Monatsbl Augenheilkd, 1976 Jan, 168(1), 134 - 7
{Gas gangrene panophthalmitis (author's transl)}; Kessler S et al.; Gas gangrene panophthalmitis is a rare condition of penetrating injury to the globe . The infecting organism is usually Clostridium perfringens . Characteristic symptoms are a brawny swelling of the lids, marked chemosis, coffee-coloured discharge, hypopyon, ring abscess of the cornea, formation of gas bubbles in the anterior chamber, rise of intraocular tension and early amaurosis . Treatment consists in the evisceration or enucleation of the globe, rarely in the exenteration of the orbit . Antibiotics along (Penicillin, Tetracyclines) are insufficient . Administration of antiserum is almost completely abandoned, it is probably more dangerous than helpful . The use of hyperbaric oxygen is not indicated in cases of gas gangrene panophthalmitis . Extraocular extension of the infection and its danger for the individual is prevented by well-times surgical procedure.

Appl Environ Microbiol, 1976 Jan, 31(1), 83 - 90
Characterization of anaerobic heterotrophic bacteria isolated from freshwater lake sediments; Molongoski JJ et al.; Strict anaerobic culture techniques were used to quantitatively and qualitatively evaluate the anaerobic heterotrophic bacteria present at the sediment-water interface of hyperutrophic Wintergreen Lake (Augusta, Mich.) . Anaerobic plate counts remained constant from March through December, 1973, ranging from 2.4 X 10(6) to 5.7 X 10(6) organisms/g (dry weight) of sediment . The isolatable bacteria represented a small percentage of the total microbial community, which was shown by direct microscopic counts to be 2.0 X 10'' organisms/g (dry weight) of sediment during June and July . Bacteria of the genus Clostridium dominated the isolates obtained, accounting for 71.8% of the 960 isolates examined . A single species, Clostridium bifermentens, comprised 47.7% of the total . Additional bacterial groups and the percentage in which they were isolated included: Streptococcus sp . (10.8%), unidentified curved rods (9.5%y, gram-positive nonsporing rods (5.6%), and motile gram-negative rods (1.9%) . Temperature growth studies demonstrated the ability of all the isolates to grow at in situ sediment temperatures . Gas-liqid radiochromatography was used to determine the soluble metabolic end products from {U-14C}glucose and a U-14C-labeled amino acid mixture by representative sedimentary clostridial isolates and by natural sediment microbial communities . At in situ temperatures the natural sediment microflora produced soluble fermentative end products characteristic of those elaborated by the clostridial isolates tested . These results are considered strong presumptive evidence that clostridia are actively metabolizing in the sediments of Wintergreen Lake.

Appl Environ Microbiol, 1976 Jan, 31(1), 25 - 8
Rapid determination of dipicolinic acid in the spores of Clostridium species by gas-liquid chromatography; Tabor MW et al.; A gas-liquid chromatographic procedure has been developed to quantitate dipicolinic acid in bacterial spores . The culture, washed from a plate, was hydrolyzed with acid containing the internal standard, pyridine-2,4-dicarboxylate, and then extracted into methyl isobutyl ketone . The internal standard and dipicolinic acid were then extracted into a small volume of trimethylphenylammonium hydroxide . Injection of the resultant quaternary ammonium salts into a gas chromatograph yielded, via thermal decomposition, the methyl ester derivatives of the dipicolinic acid and the internal standard . The amount of dipicolinic acid in the sample was determined from a standard curve . The method was sensitive to 100 ng of dipicolinic acid per sample and was 1,000 to 5,000 times more sensitive than the commonly used methods . Preparation of the sample required less than 1.5 h and less than 15 min of the analyst's time.

Mikrobiologiia, 1976 Jan-Feb, 45(1), 28 - 32
{Hydrogenase activity of different strains of Clostridium butyricum}; Gogotov IN et al.; Extracts of the cells of Clostridium butyricum, strains MO-1, USA, TM-7B, WS-6K, and C . pasteurianum 038 absorb hydrogen in the presence of its different acceptors; hydrogen is absorbed most actively by the cell extracts of C . pasteurianum 038 . The cell extracts of all studied strains of Clostridium liberate hydrogen from reduced methylviologene, benzylviologene, and azocarmine . The rate of this process in the presence of reduced methyl-viologene was highest in the strains of C . butyricum, TM-7B, and MO-1 . Hydrogenases of all studied Clostridium strains are susceptible to the action of heat and oxygen.

Ann Ist Super Sanita, 1976, 12(2-3), 142 - 69
{Microbiological methods of mineral water analysis}; Merino JR; The Spanish legislature has announced the required microbiological standards for bottled mineral waters . These requirements were the subject of "Decree 607/13th March 1975" and they are the following: -A total absence of parasites and pathogen microorganisms . -A total account of Fungus: absence in 100 ml . -A total account of coliforms: absence in 100 ml . -E . coli: entire absence in 100 ml . -A total account of Streptococcus, absence in 100 ml . -A total account of Clostridium sulphite reducing: absence in 100 ml . -A total absence of Pseudomonas aeruginosa in 100 ml . Several technical methods which have been adopted in Spain for the microbiological investigation of mineral waters have been described . For the first time the Spanish legislation has specified the total absence of Pseudomonas aeruginosa in mineral waters and techniques wich reveal the differentiation between Pseudomonas aeruginosa and other Pseudomonas by fluorescent pigments . The confirmation of diagnosis must be to carry out by the Director of National Reference Centre of Health.

Vet Med Nauki, 1976, 13(1), 53 - 8
{Normal status of Bac . cereus and Cl . perfringens in the body of healthy slaughter animals}; Milev M; Studied were cattle, pigs, sheep, and young calves intended for slaughter . The experiments were carried out under productional conditions, strictly observing the routinely adopted practice of preslaughter handling . The blood of the animals was sampled prior to slaughter . Samples from the meat (musculature), spleen, kidneys, liver, mesenterial and body lymph nodes as well as feces were taken immediately after slaughter . It was established that Bacillus cereus and Clostridium perfringens were almost lacking in the musculature, mesenterial and body lymph nodes and in some of the parenchymal organs . These organisms were chiefly found in the intestinal tract . Eighty-three up to 100 per cent of the isolated cultures of Cl . perfringens and from 62.5 up to 100 per cent of the isolated cultures of Bac . cereus originated from the feces.

Dev Biol Stand, 1976, 32, 193 - 201
Antitoxin responses to Clostridium botulinum vaccines types C and D in guinea pigs; Mathews AG; In guinea pigs, the type C and/or type D antitoxin responses to a single dose of a bivalent or monovalent Cl . botulinum vaccine increase markedly between the fourth and ninth week after injection and still increase markedly by the ninth week . For type C, a similar pattern has been found in cattle . Antigens of types C and D mutually interfere with the antitoxin responses in guinea pigs . Graded doses of vaccine arouse graded antitoxin responses in guinea pigs . Stability trials of vaccines have emphasized the unsatisfactory nature of an absolute-response type of assay, rather than revealing any loss of potency during storage . Attention is drawn to the need for a graded-response type of assay in which vaccines under test are compared with a reference vaccine.

Dev Biol Stand, 1976, 32, 175 - 83
Serological classification and typing of Clostridium botulinum; Gimenez DF; Serological classification of Cl . botulinum, based on the antigenic structure of the toxins produced, is distinguished by the behaviour of the toxin-antitoxin mixtures in in vivo neutralization tests . Observations on the dissimilarity between strains within types, the behaviour of antitoxins in the cross-neutralization tests, the established concepts of the antigenic structure of the toxin types and the lack of a standard methodology for typing, have led to the definition of the terms efficiency, type, subtype, intratypic serological variant (ISV), and degree of serological homology . These definitions are primarily applied to typing and to the establishment of the taxonomic serological categories of type, subtype and ISV . In the case of antitoxin standardization, the accepted standard methods must be followed.

Scand J Gastroenterol, 1976, 11(5), 437 - 46
Anaerobic and aerobic bacteriological studies in biliary tract disease; Nielsen ML et al.; Thirty-eight patients without biliary tract disease, 68 patients with gallbladder stones or acalculous chronic cholecystitis, 30 patients with common duct obstruction due to stones or strictures, and 28 patients with common duct obstruction due to tumors or chronic pancreatitis were investigated . Gallbladder bile or common duct bile was cultured anaerobically and aerobically . Anaerobic culture procedures were based on the use of a glove-box and prereduced, anaerobically sterilized media . Transportation of samples was based on evacuation of atmospheric air with oxygen-free carbon dioxide and transportation-time less than 30 minutes . Patients with biliary tract and patients with common duct obstruction not due to stones or strictures did not yield bacteria on culture . Twenty-two per cent of patients with disease of the gallbladder but normal common ducts and 87% of patients with common duct stones or strictures yielded bacteria on culture of the bile . Among 41 patients with infected bile, 39% yielded anaerobic bacteria, either in pure culture (12%) or in mixed culture with anaerobic bacteria (27%) . B . fragilis and other non-sporing anaerobic bacteria were most frequently isolated, whereas Clostridium species were found in three patients only.

Infect Immun, 1976 Jan, 13(1), 172 - 9
Indigenous microorganisms prevent reduction in cecal size induced by Salmonella typhimurium in vaccinated gnotobiotic mice; Tannock GW et al.; Germfree CD-1 mice challenged by the oral route with Salmonella typhimurium had ceca that were abnormal in appearance and reduced in size compared to those of germfree controls . Similarly, germfree mice injected with heat-killed S . typhimurium or gnotobiotes associated with three indigenous microbes (Lactobacillus, Bacteroides, Clostridium), and subsequently challenged with S . typhimurium also had small ceca . By contrast, gnotobiotic mice that had been both injected with the heat-killed S . typhimurium and associated with the three indigenous microbes before challenge with S . typhimurium had ceca similar in size and appearance to germfree mice . Thus, indigenous microorganisms could interfere with the mechanism by which the pathogen induced the decrease in cecal size, but could do so only in mice injected with heat-killed bacteria . This phenomenon suggests synergism between the interference effected by the indigenous bacteria and the resistance mechanisms of the animal.

Vet Med Nauki, 1976, 13(10), 91 - 5
{Demonstration and type determination of Clostridium perfringens isolated from meat semipreserves}; Popova V; Investigated were a total of 92 samples of pasteurized cans of different batches . Six strains of Clostridium perfringens were isolated from a sample of pasteurized ham . To demonstrate a toxin and present its type differentiation a strain was investigated with the use of dermonecrotic test after Williams along with the cross toxin neutralization after the internationally accepted method, and the neutralization of the lecithinase activity after Williams . The use of native toxins at the type differentiation of slightly toxigenic strains of Clostridium perfringens isolated from food products proved inappropriate . The production of dry toxin and the application of cross toxinneutralization proved to be most suitable in the typing of such strains . The type differentiation carried out as described above showed that the studied strain belonged to type A.

Arch Exp Veterinarmed, 1976, 30(6), 903 - 12
{Experimental reproduction of necrotic enteritis in the chicken . 1 . Mono- and polyinfections with Clostridium perfringens and coccidia with reference to cage managemen}; Balauca N; Experimental mono-infection and poly-infection, using vegetative germs of a CL . perfringens Type A strain 1663 and its toxin as well as E . acervulina, necatrix or mitis oocysts, were applied to 100 SPF chicken aged seven days . While clostridial or coccidial mono-infection did not cause any loss and only unspecific pathologic-anatomic changes, polyinfection was accompanied by diarrhoea and slower weight increase, with 55.4 per cent of the chicken having been lost three to nine days from the outbreak of the infection . The pathologic-anatomic findings recorded from the chicken with poly-infections included haemorrhagic, ulcerative, and necrotic intestinal inflammations, primarily in the small intestin . The results of these experimental infections indicated a possible correlation between CL . perfringens and coccidia in the pathogenesis of necrotic enteritis.

Dev Biol Stand, 1976, 32, 85 - 9
An international serotyping system for Clostridium perfringens (welchii) type A in the near future; Stringer MF et al.; The importance of Clostridium perfringens (welchii) type A as an agent of clinical infection in man is well known and, more recently, its role as a cause of human food poisoning has become well recognized . There has been a growing awareness of the need for a system of finer subdivision within the type A group for better understanding of the epidemiology of Cl . perfringens in infections and food poisoning . A set of antisera used for typing and developed in the Food Hygiene Laboratory over a period of some years has proved to be of considerable value in this respect; it is now being integrated with similar sets of antisera prepared independently in the USA and Japan to form a comprehensive and internationally workable serotyping scheme . Negotiations regarding commercial production of the main type antisera are now in progress . It is hoped that serotyping facilities will be available in the near future to workers and institutions concerned with Cl . perfringens type A.

Dev Biol Stand, 1976, 32, 77 - 83
Solid-phase radioimmunoassays for quantitative antibody determination of bacterial exotoxins . Measurement of Clostridium perfringens type D epsilon antitoxin; Bernath S; Two reversed solid-phase radioimmunoassays have been developed for quantitative determination of antibodies against Clostridium perfringens type D epsilon toxin . 125I labelled prototoxin was used in the bromoacetyl cellulose-bound antibody method and in the antibody coated tube method . The procedures are based on the competition for 125I labelled prototoxin between the insoluble antibodies and the antibodies present in the unknown sample . The radioactivity bound to solid-phase is in inverse ratio to quantity of measured antibodies . The antibody values which can be detected are in the range of 0.004 IU/ml of investigated serum . The methods allow the rapid and inexpensive screening of large groups of vaccinated sheep, and are very suitable for measuring small amounts of Cl . perfringens D epsilon antibodies with a small experimental error.

Dev Biol Stand, 1976, 32, 45 - 61
{Standardization of the control technics of Clostridium perfringens vaccine . Use of a reference vaccine}; Bousicaux A; Control of Cl perfringens toxins . It is possible to employ a standardized technique for the control of the toxins of Cl . perfringens . Titres of LD50 per ml for the mouse and L+/50 per ml may be expressed with a confidence limit of 1.5 . The assay of toxins by L+/50 (relating to their antigenic value) permits the establishment of standards by which it is possible to obtain a satisfactory vaccine with a high degree of certainty . 10 L+/50 per dose for Cl . perfringens A 20 L+/50 per dose for Cl . perfringens B 180 L+/50 per dose for Cl . perfringens C 50 L+/50 per dose for Cl . perfringens D Control of vaccine values . Technique adopted : Determination of antibodies in the rabbit following vaccination . The confidence limit of the antitoxin titre is similarly equal to 1.5 when P = 0.05 . It would not appear to be necessary to establish minimum levels of antibodies to confer protection; it seems nevertheless that the results are influenced by considerable variations attributable to the animals used for the most part and that a reference vaccine is required . Freeze-dried vaccine . There is a natural antigenic activity in the rabbit which retains a considerable part of its antigenic properties after maintenance for one week at +37 degrees C . In our view this would be suitable for a reference vaccine.

Dev Biol Stand, 1976, 32, 31 - 4
Production and standardization of polyvalent Clostridium perfringens vaccine in Iran; Ardehali M et al.; In this presentation an account of the large-scale production and the standardization of polyvalent Clostridium perfringens vaccine in Iran is described . Over 20 million doses of this vaccine are produced and utilized in Iran every year . Certain modifications have been made to the culture media used in our laboratory for the production of this vaccine in order to bring down its cost . The prepared vaccine is highly immunogenic as determined by the laboratory examination on the quality of the vaccine according to British Veterinary Codex and the field reports.

Dev Biol Stand, 1976, 32, 245 - 50
The use of a multicomponent standard for clostridial vaccines; Gray AK; A multicomponent clostridial vaccine in being developed with the objective of acting as a standard preparation for the control of vaccines containing one to eight components . It is intended initially for use in monitoring those components which can be assessed by antitoxin response assays in rabbits . The components involved, therefore, are the principle toxoids of Clostridium (perfringens) welchii types B, C and D, Cl . oedematiens type B, Cl . septicum and Cl . tetani . The proposed standard also contains the toxoid of Cl . oedematiens type D and an anaculture of Cl . chauvoei . It is hoped later to develop the use of the two latter components as standards in other test systems . The aim of establishing the standard was to provide a means of controlling inter-laboratory variation, especially that due to animals.

Can J Comp Med, 1976 Jan, 40(1), 53 - 9
Necrotic enteritis in broiler chickens . III . Reproduction of the disease; Long JR et al.; Lesions typical of necrotic enteritis could be produced experimentally in from 11-26% of broiler chickens consuming feed containing approximately 10(7) Clostridium perfringens per gram . Highest mortality was produced using isolates from field cases of necrotic enteritis which were reisolated from experimental cases in the laboratory . Penicillin in the drinking water at 100,000 I.U./litre completely prevented mortality whereas chloramphenicol at 110 mg/litre delayed the onset and reduced the number of deaths compared to controls.

Mech Ageing Dev, 1976, 5(6), 443 - 6
Change in the stability of tendon collagen during ageing in the reptile, Calotes versicolor . A brief note; Panigrahy GK et al.; Age related changes in the digestibility of tendon collagen of the garden lizard, Calotes versicolor, were traced using type I collagenase from Clostridium histolyticum . Collagen from older lizards showed less susceptibility for collagenase digestion than the younger lizards suggesting increased number of cross-linkages comparable with mammals.

Adv Exp Med Biol, 1976, 74, 68 - 82
The iron-sulfur centers and the function of hydrogenase from Clostridium pasteurianum; Chen JS et al.; Hydrogenase from C . pasteurianum is an iron-sulfur protein containing at least two tetrameric iron-sulfur centers . Information on the structure of the remaining iron atoms must await future investigation . Although the EPR spectra of dithionite-reduced hydrogenase and eight-iron Fd showed some similarity, the CD spectra clearly indicated a difference . The tetrameric iron-sulfur centers of hydrogenase were shown to undergo redox changes when hydrogenase was oxidized or reduced . However, no evidence is now available to support a role for the tetrameric Fe-S centers, responsible for the EPR spectrum A, as the primary site for H2 binding and activation . Because we have found that the {Fe4S4(SR)4}-containing ferredoxins do not have hydrogenase activity, it is conceivable that the additional iron atoms and/or certain amino acid residues of hydrogenase also contribute to the unique catalytic properties of this enzyme . Chemical synthesis of Fe-S clusters with different peptide environments and with hydrogenase function would lead to the identification of these functional groups . X-ray diffraction studies on hydrogenase will certainly complement the other approaches . Knowledge of the structure of the active site of hydrogenase will certainly accelerate research into: (1) the synthesis of a stable catalyst to replace hydrogenase in systems designed to produce H2 by coupling this catalyst to a photoreducing system; and (2) the elucidation of the active sites of more complicated iron-sulfur enzymes such as nitrogenase.

Int J Pept Protein Res, 1976, 8(2), 155 - 65
The effect of calcium chloride on the activity and inhibition of bacterial collagenase; Head DL et al.; The enzymatic behavior and inhibition patterns of collagenase of Clostridium histolyticum in the presence of 0.5 M and 3.4 mM CaCl2 have been examined viscosimetrically . The more concentrated salt was found to enhance the rate of digestion of calfskin collagen when either measured viscosimetrically or colorimetrically by trinitrobenzenesulfonate . However, the rate of digestion of calfskin gelatin is unaffected by 0.5 M CaCl2 as determined colorimetrically . Calcium chloride also proved to have a marked effect on the inhibitory behavior of a series of imidazole compounds . Histidine (10mM) is about three-fold more effective as an inhibitor in 0.5 M CaCl2 than in 3.4 mM CaCl2, whereas a reverse effect is true for histamine, Imidazolylpropionate (10mM) was only weakly inhibitory (16%) in 0.5 M CaCl2 and not at all in 3.4 mM CaCl2 . Inhibition by 10 mM imidazole was not detectable . These observations may be useful in the design of inhibitors for tissue collagenases which share a number of common characteristics with the bacterial enzyme.

Physiol Bohemoslov, 1976, 25(2), 155 - 8
Pregnancy interrupting effects of some bacterial toxins; Sechser T et al.; PIP: Pregnancy interrupting effects of some bacterial toxins were studied in mice . Embryotoxic properties of Shigella dysenteriae and Clostridium perfringens toxins, Escherichia coli endotoxin, Vibrio cholerae, and Escherichia coli interotoxins were compared up to Day 18 of pregnancy following injections on Day 6, 8, or 13 of pregnancy . Escherichia coli endotoxin caused embryotoxic effects in all stages of pregnancy while Escherichia coli enterotoxin, Vibrio cholerae enterotoxin, and Shigella dysenteriae toxin were most effective mainly at earlier stages of pregnancy . Histological studies revealed indirect embryotoxicity at later stages by placental damage . The main site of damage was the fetal side for Shigella dysenteriae toxin and the maternal side for Escherichia coli endotoxin, enterotoxin, and cholera toxin . Clostridium perfringens toxin had no embryonic effect .

Am J Med Sci, 1976 Jan-Feb, 271(1), 59 - 63
Case report . Clostridium perfringens septicemia with detection of phospholipase C activity in the serum; Moore A et al.; Clostridium perfringens septicemia with massive hemolysis is described in an elderly woman with refractory anemia . This case is highly unusual in that (1) the diagnosis was made during life by discovery of the organisms on a routine peripheral blood film, and that (2) phospholipase C activity was detected in the serum.

J Bacteriol, 1976 Jan, 125(1), 211 - 9
Spin-labeling studies on the lipids of psychrophilic, psychrotrophic, and mesophilic clostridia; Finne G et al.; Spin-labeling studies were conducted to elucidate the viscosity and phase transition temperatures of lipids isolated from psychrophilic, psychrotrophic, and mesophilic clostridia . Electron spin resonance spectroscopy indicated that the lipids, for all the growth temperatures tested, were in a fluid state and from 13 to 24 C higher than the corresponding lipid transition temperatures . When the organisms were grown at different temperatures, a psychrotropic and two mesophilic clostridia were shown to be able to adjust their lipid-phase transition temperature to the growth temperature . A psychrophilic Clostridium strain, when grown at different temperatures, synthesized lipids that had the same phase transition temperature . It is suggested that this lack of growth temperature-inducible regulation of lipid-phase transition temperature may be a molecular determinant for the psychrophily of this organism . It is proposed that the growth temperature range of an organism is dependent upon the ability of the organism to regulate its lipid fluidity within a specific range.

Vox Sang, 1976, 31(1), 64 - 6
Acquired B antigen disappearance by in vitro acetylation associated with A1 activity restoration; Gerbal A et al.; The chemical acetylation of RBC bearing the acquired B antigen led to the disappearance of the agglutinability by anti-B and restored the A1 specificity . The same results are obtained using RBC transformed in vitro by a Clostridium Tertium filtrate, where a deacetylase was reported.

Acta Biol, 1976, 27(2-3), 107 - 17
The fermentative production of acetone-butanol by Clostridium acetobutylicum; Fouad M et al.; Fourteen different media were used in the fermentative production of acetone-butanol . The highest total yields were achieved in medium I . Potato starch and soluble starch were suitable as carbon sources . The best concentrations of potato starch and soluble starch were 500.0 and 10.0 g/l, respectively . Peptone was the most favourable nitrogen source . The best concentration of peptone was 4.0 g/l . Calcium carbonate in 3.6 g/l acted as buffering agent in the fermentation process . The best initial pH value of the fermentation medium was 6.0 . The optimum temperature was 32--33degreesC . The fermentation process required 120 h to obtain maximum yields of acetone-butanol.

Appl Environ Microbiol, 1976 Jan, 31(1), 49 - 52
Inhibition of Clostridium perfringens by heated combinations of nitrite, sulfur, and ferrous or ferric ions; Asan T et al.; Heating mixtures of sodium nitrite, cysteine, and either ferrous sulfate or ferric chloride at 121 C for 20 min at pH 6.5 or 6.3 produced a potent inhibitor of Clostridium perfringens vegetative cells and spores when added to previously heat-sterilized fluid thioglycolate medium . When the mixtures containing FeSO4 at pH 5.2 or FeCl3 at pH 2.7 were heated, the inhibitory effect was not produced . These responses seem to eliminate the possibility that cysteine nitrosothiol is the agent responsible for the heated-nitrite inhibition known as the Perigo effect . The variable pH responses also cast doubt upon the role of the black Roussin salt as the agent of the Perigo effect.

Experientia Suppl, 1976, 26, 237 - 48
Properties of enzymes from Clostridium thermoaceticum and Clostridium formicoaceticum; Ljungdahl LG et al.; Methylenetetrahydrofolate dehydrogenase from C . thermoaceticum and C . formicoaceticum have been purified to homogeneity and compared . The two enzymes are very similar physically, chemically, and kinetically, but he C . thermoaceticum enzyme has a higher thermostablility, which is an intrinsic property of the protein . Formate dehydrogenase enzymes from both bacteria require selenite and tungstate for formation and these enzymes also appear to have similar properties, although the C . thermoaceticum is stable at 70 degrees C for more than one hour . Acetate kinase from C . thermoaceticum appears to be under metabolic control . It can be concluded that enzymes from C . thermoaceticum, although they are more thermostable, are very similar to corresponding enzymes from mesophilic organisms.

C R Acad Sci Hebd Seances Acad Sci D, 1975 Dec 22, 281(24), 2033 - 6
{The mode of action of bacteriocin N5 purified from Clostridium perfringens}; Ionesco H et al.; The purified bacteriocin N5 produced by Clostridium perfringens BP6K inhibited simultaneously the syntheses of DNA, RNA and protein in sensitive cells, without DNA degradation . Bacteriocin N5 inhibited the accumulation of leucin and caused the exit of the previously accumulated amino acid . The effects of bacteriocin N5 are very similar to those observed for colicins E1, K, A and I.

Eur J Biochem, 1975 Dec 15, 60(2), 467 - 76
Nitrogenase from Azotobacter chroococcum . Purification and properties of the component proteins; Yates MG et al.; 1 . A large-scale purification of the nitrogenase components from Azotobacter chroococcum yielded two non-haem iron proteins, both of which were necessary for nitrogenase activity and each had a specific activity of approximately 2000 +/- 300 nmol of acetylene reduced/mg protein per min in the presence of sautrating amounts of the other . This procedure freed the Mo-Fe protein from a protein contaminant which had an electron paramagnetic resonance signal at g = 1.94 . 2 . Both proteins were purified to homogeneity as determined by disc gel electrophoresis and ultracentrifugal analysis . Both proteins were oxygen-sensitive but not cold-labile . Ultracentrifugal analysis indicated that both proteins dissociated to a slight degree at concentrations below 2 mg/ml . 3 . The larger of the two proteins had a molecular weight of 227 000 and contained 1.9 +/- 0.3 atoms of Mo, 23 +/- 2 atoms of Fe, 20 +/- 2 acid-labile sulphide and 47 tryptophan residues/mol . The protein consists of 4 subunits of mol . wt 60 000 (approx.) . The reduced protein showed electron paramagmetic resonance signals at g = 4.29, 3.65 and 2.013 but not in the area of g = 5 to 6 . Upon oxidation abosrbance increased throughout the visible region of the ultraviolet visible spectrum, with a maximum difference between oxidised and reduced protein occurring at 430 nm . 4 . The smaller protein had a molecular weight of 64 000 and contained 4 g-atoms of Fe and 4 acid-labile sulphide groups/mol but no tryptophan . It had two subunits of mol . wt 30 800 . The reduced protein showed electron paramagnetic resonance signhe protein retained almost full activity after oxidation with phenazine methosulphate . The ultraviolet visible spectrum of oxidised protein was clearly different from that of the oxygen-inactivated protein: it had a sharp peak at 269 nm and a broad absorbance between 340 and 470 nm with a maximum difference between oxidised and reduced forms at 430 nm . Oxygen-inactivated protein showed a sharp peak at 277.5 nm and broad peaks from 305 to 360, 400 to 425 and 435 to 475 nm . 5 . Amino acid analyses of both proteins showed that most common amino acids were present with a preponderance of acidic residues . Analyses of compositional relatedness showed that the nitrogenase proteins from A . chroococcum were most closely related to those from A . vinelandii and least so to those from Clostridium pasteurianum.

Biochemistry, 1975 Dec 2, 14(24), 5257 - 60
Activation of acetyl- and butyrylcholinesterase by enzymatic removal of sialic acid from intact neuroblastoma and astroblastoma cells in culture; Stefanovic V et al.; Removal of sialic acid from intact mammalian nervous system cells in tissue culture is accompanied by an immediate increase in cellular cholinesterase activity . Treatment of hamster astroblast cells (clonal line NN) and mouse neuroblastoma cells (clonal lines S21, N18, and N115) for brief periods with a low level of Clostridium perfringens sialidase, 5 X 10(-3) units/ml, removed 1-15 mug of sialic acid per mg of cell protein and brought about a large increase in v0 and Vmax of cellular acetylcholinesterase (EC 3.1.1.7) . Butyrylcholinesterase (EC 3.1.1.8) activities also increased upon careful enzymatic removal of cellular sialic acid, and cells with characteristically low butyrylcholinesterase activity, e.g., adrenergic clonal line N115 neuroblasts displayed relatively high activity after treatment with sialidase . These findings open the possibility that adaptive regulation of cholinesterases in mammalian cells may be mediated rapidly through changes in their sialic acid content.

Zentralbl Bakteriol {Orig A}, 1975 Dec, 233(4), 542 - 52
{Bacteriocins of Clostridium septicum (author's transl)}; Schallehn G; 17 strains of Clostridium septicum were examined for bacteriocin production . Two bacteriocin-producing strains were detected by the stab culture method and the spot test using the supernatant fluids of the cultures . The bacteriocin of Cl . septicum Ovinus was active against all other strains of Cl . septicum without inhibiting the donro strain (Table 2) . This bacteriocin and one indicator strain (Wex) were chosen for further study . The bacteriocin, which was spontaneously produced during the logarithmic growth phase of the bacteriocinogenic strain Ovinus (Fig . 1) was not inducible with high temperatures or Mitomycin C . It was resistent to chloroform, but sensitive to heat (Fig . 4, 5) and proteolytic enzymes (Table 4) . Its MG is about 20000 (Fig . 3).

Appl Microbiol, 1975 Dec, 30(6), 964 - 9
Growth potential of Clostridium botulinum in fresh mushrooms packaged in semipermeable plastic film; Sugiyama H et al.; Fresh mushrooms (Agaricus bisporus) were inoculated in the stem, gill, or cap with Clostridium botulinum spores . They were placed with uninoculated mushrooms in paper board trays, which were then covered and sealed in a polyvinyl chloride stretch film to simulate prepackaged mushrooms available at retail stores . When incubated at 20 C, botulinum toxin could be detected as early as day 3, or 4, when the mushrooms still appear edible . Mushrooms inoculated in the stem with 1,000 type A spores frequently became botulinogenic; higher spore levels were needed if gills or caps were inoculation sites . Type B spores were less apt to produce toxic mushrooms . Respiration of the fresh mushrooms used up O2 more rapidly than could enter through the semipermeable wrapping film, so that the equilibrium O2 concentration became low enough for growth of C . botulinum . Inoculated mushrooms did not become botulinogenic when held at 4 C.

J Hyg (Lond), 1975 Dec, 75(3), 371 - 9
Clostridium botulinum in the lakes and waterways of London; Smith GR et al.; Mud samples collected during 1974 from a large proportion of the lakes and waterways of London were examined for Clostridium botulinum . Of 69 such sites, 50 (72.5%) contained at least one type of the organism . Of the 50 positive sites, 31, 12, 1 and 10 contained, respectively, types B, C, D and E . Most of the demonstrations of type B required trypsinization of culture filtrates . An examination of 7 lakes in Edinburgh, made for the purpose of comparison, showed that 4 contained type B and one type C . An analysis of the results gave quantitative information on the value of (1) resampling apparently negative lakes, (2) the use of both heated and unheated culture inocula, and (3) trypsinization of culture filtrates.

Proc Natl Acad Sci U S A, 1975 Dec, 72(12), 4854 - 8
NH---S hydrogen bonds in Peptococcus aerogenes ferredoxin, Clostridium pasteurianum rubredoxin, and Chromatium high potential iron protein; Adman E et al.; Results from refinement of the crystal structures of P . aerogenes ferredoxin and C . pasteurianum rubredoxin determined by x-ray diffraction show that there are 15-18 NH---S bonds in the former and six in the latter with lengths in the range 3.1-3.9 A . Earlier tritium exchange experiments are consistent with the presence of these hydrogen bonds in the ferredoxin structure and show that more peptide hydrogen atoms are available for exchange in apoferredoxin than in intact ferredoxin . Four types of NH---S bonds are observed and two of these are geometrically similar to the two types of 3(10) NH---O bonds . The existence of more NH---S bonds in ferredoxin than in high potential iron protein suggests why the -2 form of the Fe4S4 cluster is preferred in ferredoxin over the -1 form found in high potential iron protein . From comparison of Cys-X-Y-Cys sequences in rubredoxin, ferredoxin, and high potential iron protein we suggest that two Cys-X-Y-Cys-Z sequences, where Z may have conformation angles similar to glycine, are required to make a one-iron cluster, no more than one Cys-X-Y-Cys-Z-Gly sequence is required to form a Fe2S2 ferredoxin, and a Cys-X-Y-Cys-Gly sequence where Y has a conformation such that the cysteines bond to different iron atoms is necessary to form the tetrameric cluster.

Jpn J Exp Med, 1975 Dec, 45(6), 433 - 8
Clostridium perfringens exotoxins . III . Binding of theta-toxin to erythrocyte membrane; Hase J et al.; When Clostridium perfringens theta-toxin was incubated with sheep erythrocytes the toxin activity disappeared before lysis, the fact of which suggests fixation of the toxin to erythrocyte membranes . 2 . Theta-Toxin lost its activity by binding to cell membranes, and the membrane constituted inhibitor of theta-hemolysis was neither a protein, a carbohydrate nor a phosphatide, but was cholesterol . From these results this report proposes that the theta-toxin binding site of erythrocytes should be cholesterol.

Proc Natl Acad Sci U S A, 1975 Dec, 72(12), 4795 - 9
On the iron-sulfur cluster in hydrogenase from Clostridium pasteurianum W5; Erbes DL et al.; Hydrogenase, purified to an average specific activity of 328 mumol of H2 evolved/(min X mg of protein) from Clostridium pasteurianum W5, was found to have 4-5 Fe and 4-5 labile sulfur atoms per molecule of 60,000 molecular weight, in contrast with earlier reports of 12 Fe per molecule . Displacement of the iron-sulfur cluster from hydrogenase by thiophenol in 80% hexamethyl phosphoramide:20% H2O yielded the Fe4S4 (thiophenyl)4 dianion according to absorption spectroscopy . Electron paramagnetic resonance spectroscopy at 12 K showed that the iron-sulfur cluster in the enzyme could be reduced by the H2 to a state (g-values of 2.098, 1.970, and 1.898) similar to that in reduced ferredoxin and could be oxidized by dichlorophenolindophenol or H+ to a state (g-values at 2.099, 2.041, and 2.001) similar to that in high potential iron-sulfur proteins . These oxidations and reductions appeared to occur within the turnover time of the enzyme . Deuterium failed to narrow the electron paramagnetic resonance signal in either state, but the competitive inhibitor carbon monoxide reversibly formed a compound with either state and substantially altered the electron paramagnetic resonance . 13CO produced a broadening of these signals, suggesting the formation of a direct CO complex with the iron-sulfur cluster . These data are consistent with a model of the active site of the enzyme in which a four-iron four-sulfur cluster is a component that can accept one or two electrons from and donate either one or two electrons to substrates, and in which the iron-sulfur cluster serves as the site of binding of gaseous ligands.

J Bacteriol, 1975 Dec, 124(3), 1621 - 3
Competitive inhibition of transformation in group H Streptococcus strain Challis by heterologous deoxyribonucleic acid; Ceglowski P et al.; Glucosylated deoxyribonucleic acid (DNA) from phages T4 and T6 competes poorly with homologous DNA causing only a slight decrease of transformation in Group H Streptococcus strain Challis . Other types of heterologous DNAs (Micrococcus luteus, Clostridium perfringens, Escherichia coli, calf thymus and non-glucosylated phage T6 DNA), in contrast to glucosylated T4 and T6 DNAs, compete with transforming DNA to the normal, high extent . These results indicate that as in transformation of Bacillus subtilis, the presence of glucose attached to 5-hydroxymethylcytosine in phage T6 DNA considerably decreases the interaction of such DNA with competent cells of the Challis strain . It also indicates that the guanine plus cytosine content of DNA is not decisive in determining its interaction with competent cells.

Biochim Biophys Acta, 1975 Dec 1, 413(2), 309 - 16
Changes in lipid metabolism and cell morphology following attack by phospholipase C (Clostridium perfringens) on red cells or lymphocytes; Allan D et al.; When intact human erythrocytes were treated with phospholipase C (Clostridium perfringens), up to 30% of the membrane phospholipids were broken down without significant cell lysis . Only phosphatidylcholine and sphingomyelin were attacked . Ceramide (derived from sphingomyelin) accumulated, but 1,2-diacylglycerol (derived from phosphatidylcholine) was largely converted into phosphatidate . Up to 12% of the cell phospholipid could be converted into phosphatidate in this way . Pig erythrocytes and lymphocytes showed a similar but smaller synthesis of phosphatidate after phospholipase C attack . Phospholipase C also caused a marked morphological change in erythrocytes, giving rise to spherical cells containing internal membrane vesicles . This change appeared to be due to ceramide and de and diacylglycerol accumulation rather than to increased phosphatidate content of the cells.

Am J Med, 1975 Dec, 59(6), 851 - 6
Necrotizing pneumonia and empyema due to Clostridium perfringens . Report of a case and review of the literature; Bayer AS et al.; Clostridia are rare causes of pleuropulmonary infections in the absence of penetrating chest injuries; only 10 previous cases have been reported from civilian practice . An additional case of a rapidly progressive, necrotizing pneumonia and empyema is reported . Clostridial pneumonia is more likely to occur in patients with underlying pleuropulmonary disease . Unlike clostridial myonecrosis, it is rarely associated with toxemia; its mortality rate is comparable to that of nonclostridial pleuropulmonary infections . Appropriate antimicrobial therapy with surgical drainage of the empyema is the treatment of choice . Among the cases reviewed, an iatrogenic cause of infection involving an invasive procedure into the pleural cavity could be identified in seven of 11 cases . Aspiration of oropharyngeal contents was the likely route of infection in three other cases . In the remaining case, bacteremic seeding of the pleural cavity was the most probable mode of infection.

Jpn J Exp Med, 1975 Dec, 45(6), 479 - 88
Lung damage caused by phospholipase C and the changes in phospholipids in the rat lung; Ito M et al.; The lipid and protein fractions of the endobronchial lavage fluid from the normal rats which contained the lung surfactant were analysed . Lecithin, the main main component of the lung surfactant, was exclusively combined with a dextran precipitable protein . This protein then behaved as beta-globulin on cellulose acetate electrophoresis or low density lipoprotein on agarose gel filtration . After administration of phospholipase C (from Clostridium Welchii), the protein content of the lavage fluid increased markedly . The amount of dextran precipitable protein also increased markedly and its properties remained the same on gel filtration after treatment with phospholipase C . The phospholipids in the lavage fluid were not affected, although the total phospholipids in the lung tissue, especially lecithin, did decrease during the 10 days after treatment . Histopathologically, an eosinophilic dense exudative fluid appeared in both the interstitium and the broncho-alveolar spaces . A large number of the alveolar lining cells disappeared and a few of them were desquamated into the alveolar spaces . Thus, the immediate destruction of the alveolar lining cells after the administration of phospholipase C resulted in interstitial pneumonia in 10 days . The significance of phospholipase in pulmonary inflammation is discussed.

Infect Immun, 1975 Dec, 12(6), 1262 - 70
Molecular construction of Clostridium botulinum type A toxins; Sugii S et al.; Two Clostridium botulinum type A toxic fractions, named large (L) and medium (M) toxins, were eluted from Sephadex G-200 . Sucrose density gradient centrifugation resolved L toxin (2.5 X 10(8) to 3.0 X 10(8) mean lethal doses per mg of N) into two fractions, 19S and 16S . The same procedure performed at pH 8resolved it into three fractions; the heavier two were both nontoxic and hemagglutinin positive, and the lightest on (7S) was toxic . M toxin (12S) (4.5 X 10(8) to 5.0 X 10(8) mean lethal doses per mg of N) was homogeneous in electrophoresis and centrifugation at pH 6 . The latter procedure performed at pH 8 dmonstrated that it dissociated into uniform 7S components . The nontoxic component of M toxin was free from hemagglutinin . M toxin alone was demonstrated in culture by sucrose density gradient centrifugation at pH 6 . Dialysis of the culture supernatant resulted in partial formation of 16S toxin . Centrifugation of the crystalline toxin in 1 MNaCl demonstrated 16S toxin only . The toxic components of L, M, and crystalline toxins were antigenically identical . The nontoxic components of the crystalline and L toxins, consisting of two distinct antigens, were antigenically identical; that of M toxin was identical with one of these two antigens.

J Bacteriol, 1975 Dec, 124(3), 1295 - 1301
Transport of molybdate by Clostridium pasteurianum; Elliott BB et al.; The transport of 99MoO42- into dinitrogen-fixing cells of Clostridium pasteurianum was investigated . Transport of molybdate in this organism is energy dependent; sucrose is required in the minimal media, and the system is inhibited by the glycolysis inhibitors, NaF, iodoacetic acid, and arsenate . The cells accumulate molybdate against a concentration gradient, and the uptake shows a marked dependence on temperature (optimum 37 C) and pH (optimum 6.0) . The rate of molybdate uptake with increasing molybdate concentrations shows saturation kinetics with an apparent Km and Vmax of 4.8 X 10(-5) M and 55 nmol/g of dry cells per min, respectively . Inhibition studies with the anions SO42-, S2O32-, WO42-, and VO32- show that SO42- and WO42- competitively inhibit MoO42- uptake (apparent Ki {SO42-} is 3.0 X 10(-5) M; apparent Ki {WO42-} is 2.4 X 10(-5), whereas S2O32- and VO32- have no inhibitory effect . Exchange experiments with MoO42- show that only a small percentage of the 99MoO42- taken up by the cells is exchangeable . Exchange experiments with WO42- and SO42- indicate that once inside the cells WO42- and SO42- cannot substitute for MoO42-.

Biochim Biophys Acta, 1975 Nov 18, 412(1), 62 - 9
Studies on epsilon-prototoxin of Clostridium perfringens type D . Physicochemical and chemical properties of epsilon-prototoxin; Habeeb AF; epsilon-Prototoxin of clostridium perfringens type D consists of one polypeptide chain of 311 amino acids with the following composition: Asp52 Thr31 Ser25 Glu28 Pro12 Gly17 Ala14 Val28 Met5 Ile15 Leu18 Tyr17 Phe8 Lys31 His2 Arg5 Tyr2 . It has no free cysteine but contains one blocked cysteine . The N-terminal as well as the C-terminal residue is lysine . The ultracentrifuge pattern gave one single boundary having S020,w = 2.15 S and Do20,w = 5.56-10(-7) cm2/s . Calculation of the molecular weight from D020,w and S020,w gave a value of 34 250 . The molecular weight determined from sedimentation equilibrium using ultraviolet optics gave a value of 33 000 +/- 1000 . On the other hand molecular weights calculated from a calibrated Sephadex G-100 column in borate buffer was 25 000 and that in sodium phosphate, ionic strength 0.2, was 27 500 . This discrepancy between values obtained in the ultracentrifuge and by gel filtration is attributed to adsorption of epsilon-prototoxin by Sephadex.

Experientia . 1975 Nov 15;31(11):1270.
Creatinine metabolism by Clostridium welchii isolated from human faeces; ten Krooden E et al.; A Clostridium welchii has been isolated from human faeces which can deaminate creatinine to N-methyl hydantoin . Evidence suggests the reaction is inducible since the rate of conversion is increased by growth of the organism in creatinine-rich media.

Appl Microbiol, 1975 Nov, 30(5), 811 - 20
Low-temperature irradiation of beef and methods of evaluation of radappertization process; Anellis A et al.; An inoculated, irradiated beef pack (1,240 cans) was conducted for the determination of microbiological safety for unrestricted human consumption . Each can contained a mixture of 10(6) spores of each of 10 strains of Clostridium botulinum (5 type A and 5 type B), or a total of 10(7) spores/can . The cans were irradiated to various doses (100 cans/dose) with 60Co gamma rays at -30 +/- 10 C, incubated at 30 +/- 2 C for 6 months, and examined for swelling, toxicity, and recoverable botulinal cells . The minimal experimental sterilizing dose based on nonswollen, nontoxic sterile cans were 2.2 less than experimental sterilizing dose based on nonswollen, nontoxic sterile cans was 2.2 less than experimental sterilizing dose less than or equal to 2.6 Mrad . Using recoverable cells as the most stringent criterion of spoilage, and assuming the conventional simple exponential (without an initial shoulder) rate of spore kill, the "12D" dose was 3.7 Mrad when estimated on the basis of mixture of 10 strains totaling 10(7) spores/can, and 4.3 Mrad if it is assumed that each can of beef contained 10(6) spores of a single most resistant strain and all of these spores were of identical resistances . However, an analysis of the data by extreme value statistics indicated with 90% confidence that the spore death rate was not a simple exponential but might be a shifted exponential (with an initial shoulder), Weibull, lognormal, or normal, with a "12D" equivalent of about 3.0 Mrad regardless of the initial spore density per can . There was an apparent antagonism between the irradiated type A and B strains in the cans . Some of the cans contained type B toxin but did not include type B viable cells . Other cans had a mixture of type A and B toxins, but a large number of these cans did not yield recoverable type B cells . However, type A viable cells could always be demonstrated in those cans containing type A toxin.

J Biol Chem, 1975 Nov 10, 250(21), 8462 - 6
Effects of strong electrolyte upon the activity of Clostridium perfringens sialidase toward sialyllactose and sialoglycolipids; Barton NW et al.; Clostridium perfringens sialidase was purified by affinity chromatography . Kinetic properties of the enzyme were examined with sialyllactose and with mixed sialoglycolipids (gangliosides) as substrates . With the latter substrate in 0.01 M Tris-acete in the absence of strong electrolyte, the pH optimum for enzymatic activity was 6.8 . Addition of strong electrolyte (0.01 to 0.10 M Nac1) to the reaction medium caused an acidic shift and a broadening of the pH optimum, Enzymatic activity at pH 5.8 rose approximately 2.5-fold; a concomitant loss of activity at pH 6.8 was also observed . The alteration of enzymatic activity caused by strong electrolyte were dependent upon changes in Vmax . Km remained nearly invariant . Thus, a reversible transition of the enzyme from a relatively inactive to a highly active form occurred as a function of strong electrolyte concentration . Determination of the pK values of the active functional groups of C . perfringens sialidase revealed that the effects of strong electrolyte were exerted upon the pKa group of the enzyme . Strong electrolyte appeared to shield unfavorable electrostatic interactions between polyanionic sialoglycolipid micelles and the enzyme molecule, thus protecting the pKa group from inactivation . In comparision with the effects of strong electrolyte upon enzymatic activity toward the sialoglycolipid substrate, those observed with the monovalent substrate, sialyllacthose, were minor . Collectively, these findings indicate that ionic environment may effectively control the activity and relative substrate specificity of C . perfringens sialidase at a given pH . Furthermore, they explain the low pH optima and skewed pH profiles previously reported for enzymatic activity toward high molecular weight substrates.

Cancer Res, 1975 Nov, 35(11 Pt 1), 2962 - 8
The reduction of N-hydroxy-4-acetylaminobiphenyl by the intestinal microflora of the rat; Wheeler LA et al.; The role of the intestinal flora in the conversion of N-hydroxy-4-acetyl-aminobiphenyl (N-OH-AABP) to 4-acetylaminobiphenyl has been examined . This reaction, which reverses the metabolic activation of the parent carcinogen, can be demonstrated in cultures of some bacteria indigenous to the intestinal microflora . These include cultures of Clostridium sp., Clostridium perfringens, Peptostreptococcus productus I, and Bacteroides fragilis ss . thetaiotaomicron and ss . vulgatus . In contrast, cultures of Lactobacillus plantarum and Escherichia coli show little or no capacity for this reaction . The reduction of N-OH-AABP is also carried out by homogenates of liver, kidney, and brain . On a weight basis, the cecal flora is considerably more active in reducing N-OH-AABP than are homogenates of tissues of the gastrointestinal tract . The cecal flora also has a greater activity for reducing N-OH-AABP than the stomach flora, an observation which may relate to the induction of tumors in the forestomach but not in the cecum of rats fed this compound . The products of the metabolism of N-OH-AABP have been compared in germ-free and conventional animals . Glucuronide conjugates of N-OH-AABP are found in the cecal contents and feces only of the germ-free rats, while 4-acetylaminobiphenyl is found in the feces only of conventional rats . These results suggest that the flora, by hydrolyzing glucuronides and reducing N-OH-AABP, may influence the level of metabolities of 4-acetylaminobiphenyl which are critical for carcinogenesis.

J Cell Sci, 1975 Nov, 19(2), 341 - 55
The perturbation of the human erythrocyte membrane by phospholipase C; Limbrick AR et al.; A study has been made of freeze-fractured preparations of erythrocyte ghosts modified by phospholipase C (Clostridium welchii) . Such membranes show a decrease in surface area of up to about 47% and lipid droplets appear on their external surface but there is no loss of protein . Freeze-fracture of maximally hydrolysed membranes exposes only very small areas of A faces and these appear particle-free . Most of the membranes are simply cross-fractured . At lower levels of hydrolysis there is more extensive exposure of A fracture faces but the particle density is less than in control preparations . If such exposed faces were representative of the whole membrane then the particle density would have been expected to increase . It is suggested either that areas of membrane with increased particle density do not fracture or that the particles revealed by freeze-fracture involve phospholipid as well as protein and are not revealed in the absence of phospholipid.

J Biochem (Tokyo), 1975 Nov, 78(5), 905 - 9
Action of bacterial collagenase on Ascaris cuticle collagen; Fujimoto D; The collagen from the cuticle of Ascaris lumbricoides was digested by Clostridium histolyticum collagenase {EC 3.4.24.3} in the presence and absence of CaCl2 . About 1.2 mumoles of amino groups per mg collagen was liberated when the digestion was performed in the presence of 5 mM CaCl2, whereas about 0.5 mumole of amino groups per mg collagen was liberated by digestion in the absence of CaCl2 . In contrast, CaCl2 influenced the extent of hydrolysis of rat tail tendon collagen only slightly . The results suggest that CaCl2 is necessary for the hydrolysis of certain regions in the molecule of Ascaris collagen and that such structures may not be present in mammalian collagens.

Appl Microbiol, 1975 Nov, 30(5), 873 - 5
Repair of heat-injured Clostridium perfringens spores during outgrowth; Barach JT et al.; Clostridium perfringens strain NCTC 8798 spores were injured by ultrahigh temperature treatment and were unable to outgrow in the presence of antibiotics used in selective enumeration media . Injured spores underwent repair in a nonselective laboratory medium and in foods.

Appl Microbiol, 1975 Nov, 30(5), 838 - 43
Inhibitor of Clostridium perfringens formed by heating sodium nitrite in a chemically defined medium; Moran DM et al.; An inhibitor of Clostridium perfringens formed when low levels of nitrite were autoclaved with a defined chemical medium . A systematic study of the medium revealed that only amino acids and mineral salts were involved in the production of this inhibitor, which was proven to be a toxic compound formed from cysteine, ferrous sulfate, and sodium nitrite . The inhibitor was compared to several known compounds . S-nitrosocysteine inhibited the test organism, but would not form in the test system in amounts large enough to explain the observed inhibition . Roussin red salt was unstable in the test system and therefore was not the inhibitor . Roussin black salt, which was also inhibitory, could form in sufficient amounts to explain the inhibition . A complex of cysteine, iron, and nitric oxide was detected in the autoclaved solution of cysteine, ferrous sulfate, and sodium nitrite; this cysteine complex did not appear to be inhibitory, however, at levels which could form in the autoclaved medium . The observed inhibition may have been due to the combined effects of sublethal concentrations of each compound.

Appl Microbiol, 1975 Nov, 30(5), 781 - 5
Differential effects of oxygen and oxidation-reduction potential on the multiplication of three species of anaerobic intestinal bacteria; Walden WC et al.; The sensitivity of three strains of anaerobic intestinal bacteria, Clostridium perfringens, Bacteroides fragilis, and Peptococcus magnus, to the differential effects of oxygen and adverse oxidation-reduction potential was measured . The multiplication of the three organisms was inhibited in the presence of oxygen whether the medium was at a negative oxidation-reduction potential (Eh of -50 mV), poised by the intermittent addition of dithiothreitol, or at a positive oxidation-reduction potential (Eh of near +500 mV) . However, when these organisms were cultured in the presence of oxygen, no inhibition was observed, even when the oxidation-reduction potential was maintained at an average Eh of +325 mV by the addition of potassium ferricyanide . When the cultures were aerated, the growth patterns of the three organisms demonstrated different sensitivities to oxygen . P . magnus was found to be the most sensitive . After 2 h of aerobic incubation, no viable organisms could be detected . B . fragilis was intermediately sensitive to oxygen with no viable organisms detected after 5 h of aerobic incubation . C . perfringens was the least sensitive . Under conditions of aerobic incubation, viable organisms survived for 10 h . During the experiments with Clostridium, no spores were observed by spore staining.

J Laryngol Otol, 1975 Nov, 89(11), 1147 - 50
Hyperbaric oxygen in anaerobic infection of the medistinum; Monies-Chass I et al.; In the last decade hyperbaric oxygen has been recognized as an important therapeutic tool in a variety of instances in which either destruction of anaerobic bacteria is urgent or an improvement in the oxygenation level is mandatory . We have used it successfully in a case of mediastinal anaerobic infection (gas gangrene) after medical and surgical measures had failed to eradicate the disease . The causative organism of gas gangrene, Clostridium perfringens (Welchii), is widely distributed . It may be cultured from the soil, house dust, human skin and faeces, etc . For this reason infection with Clostridium is practically inevitable whenever suitable conditions arise . As an anaerobic bacterium Clostridium Welchii multiplies readily in damaged tissues without contact with the air and devoid of a normal blood supply . This occurs especially in road accidents and war wounds in which broken bones and crushed muscles provide a suitable medium for the infection (Altmeier, 1965) . Sometimes this infection can occur too after abdominal or gynaecological operations (Hitchcock, 1965) . We present here a case of mediastinal gas gangrene which was caused by perforation of the oesophagus by a swallowed foreign body.

Infect Immun, 1975 Nov, 12(5), 1214 - 8
Histopathological effect of Clostridium perfringens enterotoxin in the rabbit ileum; McDonel JL et al.; Highly purified enterotoxin from Clostridium perfringens was found to have histopathological activity in the rabbit ileum . Unlike the action of cholera, Escherichia coli, and Shigella enterotoxins, epithelium was denuded from the tips of ileal villi at concentrations of the enterotoxin necessary to induce fluid accumulation in the rabbit . Whether or not this observed histopathology is essential for the diarrheal syndrome associated with Clostridium perfringens food poisoning remains unclear.

Can J Microbiol, 1975 Nov, 21(11), 1719 - 23
A comparative study on the heat stability of triosephosphate isomerase in psychrophilic, psychrotrophic, and mesophilic clostridia; Finne G et al.; The temperature stability of triosephosphate isomerase (EC 5.3.1.1) in the cell-free extracts of psychrophilic, psychrotrophic, and mesophilic Clostridium species has been investigated . Even though this enzyme was found to be heat labile in the psychrophilic isolates, no detectable loss in activity was evident when cell-free extracts were heated for 1/2 h at the maximum temperature of growth for the organisms . Two organisms, each with a maximum growth temperature of 23 degrees C, showed different heat stability of triosephosphate isomerase . However, a trend is evident that as the maximum growth temperature increases, the thermostability also increases . It is suggested that the heat liability of this enzyme is not a controlling factor in psychrophilism, but rather an adaptation to the cold habitat of these organisms.

Lancet, 1975 Nov 1, 2(7940), 861 - 3
A continuing common-source outbreak of botulism in a family; Horwitz MA et al.; In December, 1974, three cases of botulism occurred in a family; two were fatal . The first patient died after a 10-day illness without botulism being suspected . 4 days later, after a 2-day illness, the second patient was diagnosed as having botulism after a cardiorespiratory arrest; she died 3 days later . In the third patient, the only symptom was dysphagia . Clostridium botulinum type B was found in stool specimens from all three patients . Home-canned (bottled) mushrooms, which were found to contain C . botulinum type B and its toxin, were believed to be responsible for the outbreak; mushrooms were found at necropsy in the gastrointestinal tracts of both patients who died . Heat treatment of the mushrooms during canning had been inadequate.

Can J Microbiol, 1975 Nov, 21(11), 1711 - 8
Regulation and properties of an invertase from Clostridium pasteurianum; Laishley EJ; An intracellular invertase was induced in cultures of Clostridium pasteurianum utilizing sucrose as its carbon source for growth . This enzyme synthesis could be repressed by the addition of fructose of a sucrose-growing culture . In contrast, invertase activity was not affected by the addition of glucose to sucrose-growing cells and this enzyme could be induced in a glucose-metabolizing culture by the addition of sucrose . This enzyme was purified 10.5-fold over the induced lese, EC 3.2.1.26) by substrate-specificity studies . Invertase had a pH optimum of 6.5 and an apparent Km of 79.5 mM for sucrose, and required high concentration of potassium phosphate for maximum activity . Invertase was completely inactivated by a 2-min heat treatment at 60 degrees C . This enzyme was strongly inhibited by p-hydroxymercuribenzoate (pCMB) and weakly inhibited by 5,5'-dithiobis(2-nitrobenzoic acid), while cysteine could substantially reverse pCMB) inhibition, suggesting that sulfhydryl group(s) were necessary for invertase activity.

J Biol Chem, 1975 Oct 25, 250(20), 8049 - 54
The monovalent cation-induced association of formyltetrahydrofolate synthetase subunits . Kinetic and thermodynamic aspects; Harmony JA et al.; Formyltetrahydrofolate synthetase monomers are converted to catalytically active tetramers in the presence of monovalent cations . The stoichiometry of the reaction is 4M + 2C+ in equilibrium M4C2(2+) . A positive deltaS compensates for an unfavorable positive deltaH so that the overall reaction is exergonic . Both deltaH and deltaS become more positive as the temperature is increased . Association of subunits of the enzyme prepared from Clostridium cylindrosporum is second order with respect to monomer concentration, consistent with a rate-determining dimerization step . Activation parameters for this step at 20 degrees are: deltaG, 12.6 kcal mol-1; deltaH, 12.5 kcal mol-1; deltaS, -05 e.u . The rate-limiting step for the cation-dependent association of Clostridium acidi-urici monomers is believed to be a conformational alteration since first order kinetics is observed . The Eyring plot of the kinetic data obtained for the C . acidi-urici system has a sharp break at 15 degrees . Activation parameters for cation-induced association at 20 degrees are: deltaG, 21.5 kcal mol-1; deltaH, 14.0 kcal mol-1; deltaS, -26.6 e.u.

Biochim Biophys Acta, 1975 Oct 22, 403(2), 521 - 9
Regulatory responses of arginine deiminase in whole cells of Clostridium sporogenes; Venugopal V et al.; Arginine deiminase (EC 3.5.3.6) has been shown to have regulatory properties . The activity was observed to be sigmoidal with respect to substrate concentrations . Addition of histidine to the system caused the abolition of sigmoidal responses . The regulatory properties of the enzyme as well as the desensitising action of histidine could also be demonstrated with whole cell suspensions . The pH of the system also seemed to influence modulations in the enzyme.

Biochim Biophys Acta, 1975 Oct 21, 409(1), 75 - 85
Phospholipase C from Clostridium novyi type A . I; Taguchi R et al.; 1 . Phospholipase C (EC 3.1.4.3) from Clostridium novyi (oedematiens) type A was purified 2000-fold by (NH4)2SO4 precipitation, DEAE-Sephadex treatment in a batchwise system and Sephadex G-100 column chromatography . 2 . The purified preparation had a specific activity of 95 mumol per min per mg protein toward phosphatidylcholine . This preparation was free from protease, lipase and oxygen-labile delta-hemolysin . 3 . Phosphatidylcholine was hydrolyzed at the highest rate, while sphingomyelin and lysophosphatidylcholine were hydrolyzed at much lower rates . 4 . Sodium deoxycholate and divalent cations such as Mg2+ and Ca2+ were extremely effective in stimulating phosphatidylcholine-hydrolyzing activity of this enzyme . 5 . This enzyme hemolyzed horse red cells by hydrolyzing phosphatidylcholine, spingomyelin and phosphatidylethanolamine.

J Biol Chem, 1975 Oct 10, 250(19), 7842 - 7
High and low reduction potential 4Fe-4S clusters in Azotobacter vinelandii (4Fe-4S) 2ferredoxin I . Influence of the polypeptide on the reduction potentials; Sweeney WV et al.; Azotobacter vinelandii (4Fe-4S)2 ferredoxin I (Fd I) is an electron transfer protein with Mr equals 14,500 and Eo equals -420 mv . It exhibits and EPR signal of g equals 2.01 in its isolated form . This resonance is almost identical with the signal that originates from a "super-oxidized" state of the 4Fe-4S cluster of potassium ferricyanide-treated Clostridium ferredoxin . A cluster that exhibits this EPR signal at g equals 2.01 is in the same formal oxidation state as the cluster in oxidized Chromatium High-Potential-Iron-Protein (HiPIP) . On photoreduction of Fd I with spinach chloroplast fragments, the resonance at g equals 2.01 vanishes and no EPR signal is observed . This EPR behavior is analogous to that of reduced HiPIP, which also fails to exhibit an EPR spectrum . These characteristics suggest that a cluster in A . vinelandii Fd I functions between the same pair of states on reduction as does the cluster in HiPIP, but with a midpoint reduction potential of -420 mv in contrast to the value of +350 mv characteristic of HiPIP . Quantitative EPR and stoichoimetry studies showed that only one 4Fe-4S cluster in this (4Fe-4S)2 ferredoxin is reduced . Oxidation of Fd I with potassium ferricyanide results in the uptake of 1 electron/mol as determined by quantitative EPR spectroscopy . This indicates that a cluster in Fd I shows no electron paramagnetic resonance in the isolated form of the protein accepts an electron on oxidation, as indicated by the EPR spectrum, and becomes paramagnetic . The EPR behavior of this oxidizable cluster indicates that it also functions between the same pair of oxidation states as does the Fe-S cluster in HiPIP . The midpoint reduction potential of this cluster is approximately +340 mv . A . vinelandii Fd I is the first example of an iron-sulfur protein which contains both a high potential cluster (approximately +340 mv) and a low potential cluster (-420 mv) . Both Fe-S clusters appear to function between the same pair of oxidation states as the single Fe-S cluster in Chromatium HiPIP, although the midpoint reduction potentials of the two clusters are approximately 760 mv different.

Zentralbl Bakteriol {Orig A}, 1975 Oct, 233(2), 253 - 60
Studies on the cultivation of Clostridium oncolyticum M 55 . 5th communication: The influence of iron, zinc, and cobaltous ions on the growth and the kininase activity of clostridium oncolyticum M 55 ATCC 13.732; Brantner H et al.; The precipitation or iron sulfide by hydrogen sulfide excreting Clostridium oncolyticum M 55 led to further examinations in order to avoid this disturbing factor . The method should render the cultivation of Cl . oncolyticum M 55 under definite conditions and should also intensify the formation and the enrichment of the clostridial kininases . Cobaltous ions and zinc granules were found useful for cultivation, because no disturbances increased from both during sterilization and incubation . The determination of the growth cycles and the replication rates showed the usefulness of zinc for substitution of iron . Cobaltous ions did only allow a low replication rate . These results confirmed those showing the influence of chelate-forming agents on the kininase activity . Cobaltous ions were not able to exchange the central atom of the metallo-proteide . The influence of the peptone quality on the kininase activity in presence of zinc was also examined, since examinations with iron media have shown a direct connection between peptone quality and enzyme activity . The results with zinc-containing media were nearly 5-10% lower than those with iron . The results allowed also the conclusion that the kininase excreted by Cl . oncolyticum M 55 is an iron-containing metallo-proteide, whose central atom is exchangeable for zinc . The consequences of the loss of enzyme activity for the oncolytic therapy with Cl . oncolyticum M 55 will be shown in further examinations.

J Biochem (Tokyo), 1975 Oct, 78(4), 673 - 8
Characterization of two glycoasparagines isolated from the urine of patients with aspartylglycosylaminuria (AGU); Sugahara K et al.; Two major glycoasparagines (2-acetamido-N-(4'-L-aspartyl)-2-deoxy-beta-D-glycosylamines) were isolated from the urine of patients with aspartylglycosylaminuria (AGU) . They were composed of equimolar amounts of sialic acid, galactose, glucosamine, and aspartic acid . They were isomeric with respect to the position of sialic acid attachment, since they produced the same glycoasparagine on incubation with the neuraminidase from Clostridium perfringens . The structure of the resulting sialic acid-free glycoasparagine was determined as beta-Gal-(1 leads to 4)-beta-GlcNAc-Asn based on the following findings . It produced galactose on incubation with beta-galactosidase, and N-acetyllactosamine and aspartic acid on incubation with 4-L-aspartylglycosylamine amindo hydrolase.

Proc Natl Acad Sci U S A, 1975 Oct, 72(10), 4003 - 7
Determination of the iron-sulfur distances in rubredoxin by x-ray absorption spectroscopy; Shulman RG et al.; The high intensity x-ray flux from the synchrotron radiation at the Stanford Synchroton Radiation Project has been used to study the extended x-ray absorption fine structure (EXAFS) of the iron-sulfur protein Peptococcus aerogenes rubredoxin . Absorption measurements were made from 7080 eV, which is below the K-edge of iron, to about 650 eV above the edge and structure was obtained over the entire region . By means of a model iron-sulfur compound for evaluating the phase shifts, the variation of the absorption above the edge of lyophilized, oxidized rubredoxin was converted to iron-sulfur distances . The data were fitted with a least squares program to a model in which three distances R3 were kept equal and the fourth R1 was allowed to differ . The mean square error was constant over a region of this parameter space, becoming twice as large at R3 = 2.217, R1 = 2.389 and R3 = 2.268, R1 = 2.108 A . These values, which are the extreme differences allowed by the present data, are definitely closer to being equal than those found by the determination of the x-ray diffraction crystal structure of the similar protein from Clostridium pasteurianum . However, the average distance from our experiment is in excellent agreement with the average distance from the crystal structure determination . Preliminary EXAFS measurements were also made on the oxidized rubredoxin in solution at pH 7.0 . The spectra were unchanged, indicating that the average iron-sulfur distance change is less than 0.02 A . Upon reduction the average iron-sulfur bond length increased by about 0.05 A . Since the EXAFS measurements can give accurate determinations of distances in proteins both in crystals and solution, the technique should be widely applicable.

J Biochem (Tokyo), 1975 Oct, 78(4), 763 - 72
Formation of beta-methylmalate and its conversion to citramalate in Rhodospirillum rubrum; Osumi T et al.; Using a cell-free extract of Rhodospirillum rubrum, studies were made of the condensation reaction between propionyl-CoA and glyoxylate . When {14C}propionate was incubated with the extract in the presence of glyoxylate, ATP, CoA, Mg2+, and Mn2+, radioactivity was incorporated into several compounds . Two of the main products were characterized as citramalate (CMA) and erythro-beta-methylmalate (erythro-MMA) on the basis of their behavior compared with authentic samples of CMA and erythro-MMA in the following three analyses: (i) paper chromatography using two solvent systems, (ii) radio-gas chromatography on their methyl esters, and (iii) chemical conversion to readily crystallizable derivatives, that is, citramalyl chloralide for CMA, and thymine for MMA . The CMA was thought to be of L(+)-form based on the results of optical resolution with brucine and also its susceptibility to L(+)-citramalate lyase of Clostridium tetanomorphum . When the reaction was carried out with lower concentrations of the enzyme, only MMA was accumulated . However, when the reaction was allowed to proceed further after addition of higher concentrations of the enzyme and of excess semicarbazide to prevent further condensation, the amount of accumulated MMA was decreased and CMA was formed instead . Furthermore, the time course of MMA and CMA formation exhibited a pattern typical of a precursor-product relationship . From these results, it was concluded that MMA was formed by alpha-condensation between propionyl-CoA and glyoxylate, and that CMA was derived from MMA, possibly from its CoA derivative.

Am J Med Technol, 1975 Oct, 41(10), 377 - 86
Hematological findings in acute infections and septicemias; Talluto MR; This paper is a report on twelve cases of septicemia, the diagnosis made possible by medical technologists observing the degenerative morphological findings in the peripheral blood smears . This led to the finding of microorganisms, prior to the bacteriological confirmation, in an active emergency department . It also includes four case reports in which the organism was identified: a meningococcus, a pneumonococcus, an Escherichia coli, and a clostridium perfringens (welchii).

Biochem J, 1975 Oct, 151(1), 75 - 83
Physicochemical characterization of the four-iron-four-sulphide ferredoxin from Bacillus stearothermophilus; Mullinger RN et al.; 1 . A stable ferredoxin was prepared from Bacillus stearothermophilus and purified by chromatography on DEAE-cellulose and by electrophoresis . 2 . The minimum molecular weight determined from the amino acid composition was about 7900 and this was in reasonable agreement with a value of 8500 determined by polyacrylamide-gel electrophoresis . The ferredoxin contained four iron atoms and four labile sulphide groups per molecule . 3 . The optical absorption, optical-rotatory-dispersion and circular-dichroism spectra are typical of ferredoxins containing 4Fe-4S clusters . 4 . Oxidation-reduction titrations, combined with electron-paramagnetic-resonance (e.p.r.) spectroscopy, showed that the protein has a mid-point potential, at pH8, of -280 +/- 10mV, and that only one electron-accepting paramagnetic species is present . 5 . The e.p.r . spectrum of the reduced ferredoxin is more readily saturated with microwave power at low temperatures than those of the eight-iron ferredoxins, indicating that there is another mechanism of electron-spin relaxation in the latter . 6 . Mossbauer spectra of both redox states were observed over a range of temperatures and in magnetic fields . At high temperatures (77 degrees K and above) both redox states appear as quadrupole-split doublets; in the reduced state two resolved doublets are seen, suggesting appreciable localization of the additional reducing electron . 7 . The average chemical shift indicates formal valences of two Fe3+ and two Fe2+ in the oxidized state and three Fe2+ and one Fe3+ in the reduced state . However, the spectra indicate that there are differing degrees of electron delocalization over the iron atoms . 8 . At low temperatures (4.2 degrees K) the oxidized form shows no hyperfine magnetic interaction, even in an applied magnetic field, evidence that the oxidized ferredoxin is in a non-magnetic state as a result of antiferromagnetic coupling between the iron atoms . 9 . At 4.2 degrees K the reduced form shows a broad asymmetric pattern resulting from magnetic hyperfine interaction . This contrasts with the reduced ferredoxin of Clostridium pasteurianum, which shows a doublet, suggesting that in the latter there may be interaction between the two 4Fe-4S centres . 10 . In large applied magnetic fields, positive and negative hyperfine fields are seen in the Mossbauer spectra of the reduced ferredoxin, evidence for antiferromagnetic coupling between the iron atoms in the 4Fe-4S centre . The high-field spectra of the reduced ferredoxin of B . stearothermophilus are similar to those of the reduced ferredoxin of C . pasteurianum.

Ann Microbiol (Paris), 1975 Oct-Nov, 126(3), 343 - 56
{Purification and characterization of "Clostridium perfringens" BP6K-N5 strain bacteriocin N5 (author's transl)}; Wolff A et al.; The bacteriocin N5 produced by Clostridium perfringens, strain BP6K-N5, after UV irradiation induction, was purified by ammonium sulfate precipitation, followed by ion-exchange chromatography on DEAE cellulose and gel filtration on Sephadex G100 . By polyacrylamid-gel electrophoresis and immunodiffusion analysis the purified material was shown to be homogenous . The purified bacteriocin N5 is inactivated by proteolytic enzymes and by heat treatment at 50 degrees C for 15 minutes . It is a simple protein with a molecular weight of approximately 82.000 . The protein appears to be a single polypeptide chain, as no dissociation is obtained by sodium dodecyl sulfate and beta-mercaptoethanol disc-gel-electrophoresis.

Appl Microbiol, 1975 Oct, 30(4), 530 - 5
Convenient non-chromatographic assays for the microbial deconjugation and 7alpha-OH bioconversion of taurocholate; MacDonald IA et al.; We described two convenient assay methods to estimate bile acid deconjugation and bile acid bioconversion at the 7alpha-OH position by individual microorganisms grown in media containing taurocholic acid . The methods are based on (i) a selective chemical assay for taurine conjugates previously described and (ii) the use of a cell-free preparation of 7alpha-hydroxysteroid dehydrogenase from Escherichia coli to directly quantify 7alpha-OH groups . These non-chromatographic approaches have been applied to the study of three model strains of intestinal organisms, E . coli, Bacteroides fragilis, and Clostridium perfringens, grown in standard media in the presence of purified tritiated taurocholate . Assay results were confirmed by thin-layer chromatography solvent systems designed to separate conjugated from unconjugated bile acid and unmodified cholic acid nucleus from 7alpha-OH bioconversion product(s) (primarily 3alpha, 12alpha dihydroxy, 7-keto-cholanoic acid) . In addition, 7alpha-hydroxysteroid dehydrogenase activity was demonstrated in cell-free extracts of all three organisms . Of the three organisms, only C . perfringens was demonstrated to (i) deconjugate taurocholic acid, (ii) contain 3alpha-hydroxysteroid dehydrogenase activity, (iii) convert cholic acid into at least five labeled metabolites visible on thin-layer chromatography, and (iv) catalyze significant tritium exchange with water in the medium.

Appl Microbiol, 1975 Oct, 30(4), 499 - 502
Solid-phase radioimmunoassays for quantitative antibody determination of Clostridium perfringens type D epsilon toxin; Bernath S; Two reversed solid-phase radioimmunoassays were developed for quantitative determination of antibodies against Clostridium perfringens type D epsilon toxin . 125I-labeled prototoxin was used in the bromoacetylcellulose-bound antibody method and in the antibody-coated tube method . The antibody values which can be detected by the assays are in the range of 0.004 IU/ml of investigated serum . The methods allow the screening investigation of large groups of vaccinated sheep in a rapid and inexpensive way, and are very suitable for measuring small amounts of C . perfringens D epsilon antibodies with a small experimental error.

Arch Microbiol, 1975 Sep 30, 105(1), 67 - 71
Effect of cyclic AMP on catabolite repressed bacterial sporogenesis of an anaerobe; Emeruwa AC et al.; The sporulation of a high frequency sporogenic mutant of Clostridium botulinum was reduced to less than 30% in a medium containing 270 mM glucose . The repression was reversed from 30 to greater than 80% sporulation by the addition of 10(-5) or 10(-4) M cyclic 3',5'-adenosine monophosphate (cAMP) or monobutyrul cyclic AMP (B-cAMP) . No difference was observed in amount of growth with the addition of either the cAMP or B-cAMP . Glucose consumption was enhanced by the addition of either of the cyclic nucleotides and corresponding changes in pH were observed . The catabolite repression by glucose was reversed by ATP or ADP . Except for GTP, guanine nucleotides were not effective . The intracellular cyclic AMP levels were high in vegetative, sporulating and derepressed cells, but low in glucose-repressed cells . The findings suggest that the sporulation of the anaerobe was sensitive to catabolite repression which was specifically reversed by cyclic AMP.

J Biol Chem, 1975 Sep 25, 250(18), 7435 - 42
Tetanus toxin . The effect of chemical modifications on toxicity, immunogenicity, and conformation; Robinson JP et al.; Tetanus toxin has been isolated from the extract of Clostridium tetani and analyzed for purity using various methods, e.g . sedimentation velocity, gel filtration, polyacrylamide gel electrophoresis, immunoelectrophoresis, and immunodiffusion . The homogeneous toxin, characterized by a minimum lethal dose of 10 pg (18- to 20-g mouse), was judged to be of high purity . The amino acid composition was determined and found to be in good agreement with reported values for both filtrate and extract toxin . The corrected sedimentation coefficient, so20,w, was found to be 7.5, and the molecular weight was estimated to be 150,000 . These values agree closely with those reported by others . The toxin was modified using the conventional formaldehyde reaction to produce toxoid and both reductive methylation and carbamylation, which are highly specific for lysyl residues . Under certain reaction conditions, carbamylation of the toxin completely eliminated toxicity . Whereas reductive methylation yielded a high degree of conversion of lysine to dimethyllysine and monomethyllysine, the toxicity, albeit greatly reduced, was never completely eliminated . The circular dichroic spectrum of each chemically modified toxin was obtained, resolved into Gaussian components, and compared with that of native toxin, which is estimated to contain about 20% alpha helix and 23% beta structure . The far ultraviolet circular dichroic spectra of toxin and toxoid were characterized by negative extrema at 208 nm and 217 nm attributable to ordered secondary structure, and toxin also exhibited a distinct shoulder at 223 nm . Carbamylated toxin and methylated toxin were characterized by negative extrema at 210 nm and 206 nm, respectively, and both exhibited shoulders at 216 to 217 nm and 223 nm . The toxin and derivatives exhibited multiple negative extrema above 250 nm which were assigned to the various aromatic residues . There were differences in the spectra of the toxin and derivatives over the entire wavelength region, thus suggesting changes in the local environment of various chromophores . In particular, the rotational strengths of many of the bands assigned to tryptophan, tyrosine, and phenylalanine were altered in the derivatives . Also, in the far ultraviolet region of the circular dichroic spectrum, the data were suggestive of some reduction in the amount of both alpha helix and beta structure in the derivatives . However, there was no evidence of extensive conformational changes, e.g . unfolding, in the modified toxins . Presently, it is not known if the small conformational differences between toxin and toxoid are important in the loss of toxicity with the retention of immunogenicity in the derivative . The modification data are consistent with the hypothesis that separate amino acid residues are involved in toxicity and immunogenicity.

Biochim Biophys Acta, 1975 Sep 16, 406(1), 97 - 107
Relation between various phospholipase actions on human red cell membranes and the interfacial phospholipid pressure in monolayers; Demel RA et al.; The action of purified phospholipases on monomolecular films of various interfacial pressures is compared with the action on erythrocyte membranes . The phospholipases which cannot hyorolyse phospholipids of the intact erythrocyte membrane, phospholipase C from Bacillus cereus, phospholipase A2 from pig pancreas and Crotalus adamanteus and phospholipase D from cabbage, can hydrolyse phospholipid monolayers at pressure below 31 dynes/cm only . The phospholipases which can hydrolyse phospholipids of the intact erythrocyte membrane, phospholipase C from Clostridium welchii phospholipase A2 from Naja naja and bee venom and sphingomyelinase from Staphylococcus aureus, can hydrolyse phospholipid monolayers at pressure above 31 dynes/cm . It is concluded that the lipid packing in the outer monolayer of the erythrocyte membrane is comparable with a lateral surface pressure between 31 and 34.8 dynes/cm.

Biochim Biophys Acta, 1975 Sep 16, 406(1), 83 - 96
Organization of phospholipids in human red cell membranes as detected by the action of various purified phospholipases; Zwaal RF et al.; 1 . The action of eight purified phospholipases on intact human erythrocytes has been investigated . Four enzymes, e.g . phospholipases A2 from pancreas and Crotalus adamanteus, phospholipase C from Bacillus cereus, and phospholipase D from cabbage produce neither haemolysis nor hydrolysis of phospholipids in intact cells . On the other hand, both phospholipases A2 from bee venom and Naja naja cause a non-haemolytic breakdown of more than 50% of the lecithin, while sphingomyelinase C from Staphylococcus aureus is able to produce a non-lytic degradation of more than 80% of the sphingomyelin . 2 . Phospholipase C from Clostridium welchii appeared to be the only lipolytic enzyme tested, which produces haemolysis of human erythrocytes . Evidence is presented that the unique properties of the enzyme itself, rather than possible contaminations in the purified preparation, are responsible for the observed haemolytic effect . 3 . With non-sealed ghosts, all phospholipases produce essentially complete breakdown of those phospholipids which can be considered as proper substrates for the enzymes involved . 4 . Due to its absolute requirement for Ca2+, pancreatic phospholipase A2 can be trapped inside resealed ghosts in the presence of EDTA, without producing phospholipid breakdown during the resealing procedure . Subsequent addition of Ca2+ stimulates phospholipase A2 activity at the inside of the resealed cell, eventually leading to lysis . Before lysis occurs, however, 25% of the lecithin, half of the phosphatidylethanolamine and some 65% of the phosphatidylserine can be hydrolysed . This observation is explained in relation to an asymmetric phospholipid distribution in red cell membranes.

Mikrobiologiia, 1975 Sep-Oct, 44(5), 893 - 8
{Correlation of the DNA nucleotide makeup with the physiological and cytological characteristics of spore-forming anaerobic bacteria}; Duda VI et al.; The nucleotide composition of DNA from 12 studied species of anaerobic bacteria belongs to AT type, with G+C varying from 28.4 to 36.8 mole% . In the anaerobic group of Clostridium bifermentans, a correlation has been established between the nucleotide composition of DNA, the type of appendages on spores, and some physiologo-biochemical characteristics . The nucleotide composition of DNA in the spores of four anaerobic species is shifted toward GC type as compared to DNA in the vegetative cells . Data on the content of GC pairs in DNA of the spores may sometimes be of a higher taxonomic value than the corresponding evidence on DNA of the vegetative cells.

Appl Microbiol, 1975 Sep, 30(3), 420 - 3
Detection of Clostridium botulinum toxin by local paralysis elicited with intramuscular challenge; Sugiyama H et al.; Clostridium botulinum toxin can be identified by a characteristic, acute local paralysis that follows its injection into the gastrocnemius ("calf" muscle) of mice . The local botulism can be elicited with slightly less than one-tenth the toxin amount that is needed to kill mice by the intraperitoneal (i.p.) challenge route . The practical sensitivities of the intramuscular (i.m.) versus i.p . tests are about equal, however, because maximum sample volume injectable i.m . is 0.1 ml as compared to the 0.5-ml range that can be given i.p . i.m . injection of 10 or more mouse i.p . mean lethal doses causes paralysis in about 1 h, and an i.m . injection of about 0.5 i.p . mean lethal doses causes paralysis in 3 to 4 h . Toxin neutralization by homologous type of antitoxin only can be demonstrated with an incubated mixture of toxin and antitoxin . Although not as convenient as the i.p . method for routine use to detect botulinum toxin, the i.m . method has characteristics which could make it a useful supplement to the presently accepted i.p . procedure.

Hoppe Seylers Z Physiol Chem, 1975 Sep, 356(9), 1353 - 8
{Vergleichende Untersuchungen mit (14c)5-Aminolävulinat und (14c)Uroporphyrinogen (author's transl)}; Dauner HO et al.; Cell-free extracts from Clostridium tetanomorphum, a microorganism which synthesizes corrins but no heme, are capable of converting both 5-aminolevulinate and uroporphyrinogen III into cobyrinic acid . Comparative examinations with (14C)5-aminolevulinate and (14C)uroporphyrinogen yielded corresponding results . Cell-free extracts from Clostridium tetanomorphum contain uroporphyrinogen III . To obtain good radiochemical yields it is therefore necessary to use substrates of high specific radioactivity . A method for the preparation of 14C-labelled uroporphyrin I-IV with high specific radioactivity is described.

Am J Clin Pathol, 1975 Sep, 64(3), 385 - 8
Primary osteomyelitis due to an anaerobic microorganism; Isenberg HD et al.; Primary osteomyelitis in a teen-aged boy that mimicked Ewing's tumor radiologically showed small Gram-negative rods on the original smear . The organism isolated was an obligately anaerobic bacterium, finally identified as Clostridium sphenoides . This finding underlines the need for microbiologic analysis of orthopedic lesions.

J Bacteriol, 1975 Sep, 123(3), 978 - 84
Role of molybdenum in dinitrogen fixation by Clostridium pasteurianum; Cardenas J et al.; The role of Mo in the activity and synthesis of the nitrogenase components of Clostridium pasteurianum has been studied by observing the competition of Mo with its structural analogue W . Clostridial cells when fixing N2 appeared strictly dependent upon the available Mo, showing maximal N2-fixing activity at molybdate concentrations in the media of 10 muM . Cells grown in media with 3 times 10(-6) muM Mo, although showing good growth, had only 15% as much N2-fixing activity . In the presence of W the synthesis of both nitrogenase components, molybdoferredoxin and azoferredoxin, was affected . Attempts to produce nitrogenase in W-grown cells by addition of high molybdenum to the media in the presence of inhibitors of protein synthesis showed that Mo incorporation into a possible inactive preformed apoenzyme did not occur . Unlike other molybdoenzyme-containing cells, in which W either is incorporated in place of Mo to yield inactive protein or initiates the production of apoprotein, C . pasteurianum forms neither a tungsten substituted molybdoferredoxin nor an apoprotein . It is concluded that in C . pasteurianum molybdenum is an essential requirement for both the biosynthesis and activity of its nitrogenase.

J Bacteriol, 1975 Sep, 123(3), 1115 - 23
Regulation of breakdown and synthesis of L-glutamate decarboxylase in Clostridium perfringens; Cozzani I et al.; L-Glutamate decarboxylase (GAD) activity of Clostridium perfringens (ATCC 8009) cells grown in various culture conditions was investigated . Remarkable variations of GAD level occur during the growth cycle in thioglycollate broth . These changes are affected by the pH of the culture medium . Addition of alkali to the culture media results in decrease of cell GAD activity, whereas increase of enzyme level occurs only in cells growing in unbuffered media . The results indicate that the mechanism regulating the GAD levels is sensitive to the changes of pH (or buffering substances) rather than to the steady pH values . Neither repression by glucose nor induction by L-glutamate was observed . Moreover, high concentrations of the free amino acid substrate in the culture media considerably decrease cell GAD activity, owing to the buffering effect of the amino acid . The molecular mechanism supporting the variations of GAD activity during the growth cycle of the cells were investigated and tentatively related to the structural and functional properties of the pure enzyme . It is shown that the drop of GAD activity during the lag phase is due to protein breakdown . Evidence is presented suggesting a control of protein degradation by its quaternary structure . Data are also reported supporting de novo synthesis of GAD during the late logarithmic phase of cell growth . Finally, the possible role of GAD as part of the pH regulation system of C . perfringens cells is discussed in relation both to physiologic conditions of the bacterial cell and to the molecular mechanisms regulating the GAD activity in vivo.

Zentralbl Bakteriol {Orig A}, 1975 Sep, 233(1), 75 - 9
Phenotypic chain formation in Clostridium welchii by suramin; Shaikh MR et al.; Cl . welchii NCTC 6785 was grown in 1% glucose containing medium having 1% Suramin w/v . The cells grew into long chains of thirty units . This effect was not found beyond 0.5% concentration . On transfer into Suramin free medium the chains reverted to normal morphology indicating the change in morphology to be phenotypic.

Vopr Neirokhir, 1975 Sep-Oct, (5), 24 - 8
{Serologic diagnosis of brain tumors using Clostridium butyrium spores (experimental and clinical studies)}; Krechmer Kh; Spores of the Clostridium butyrium M 55 strain, introuduced intravenously, proliferate selectivity in tumours of individual with newgrowths forming bacilli and then multiply evoking the appearance of antibacillary antibodies . In persons with no tumours there emerge only antispore antibodies . Since the spores and bacilli of this strain have antigens of a different nature their antibodies are encountered separately . The finding of antibacillary antibodies following injection of spores thus serves as an indirect proof for the presence of a malignant tumour . In experiments on animals (rats) with neurogenic tumours induced with ethylnitosole urea and also in patients with brain tumours this phenomenon has been studied in detail.

Onderstepoort J Vet Res, 1975 Sep, 42(3), 91 - 8
The partial purification of Clostridium perfringens beta toxin; Worthington RW et al.; An attempt was made to purify Clostridium perfringens Beta toxin . Crude toxin prepared by ammonium sulphate precipitation of culture supernatants was purified by chromatography on Sephadex G50, Sephadex G100 and DEAE cellulose . This material, although highly purified was not homogeneous on polyacrylamide gel electrophoresis . It had a toxicity of 800 000 mouse MLDs/mg N, a typical protein absorption spectrum in the UV region, an iso-electric point of 5, 6 and the main component had a molecular mass of 42 000 +/- 2 000 (estimated by electrophoresis in sodium dodecyl sulphate containing polyacrylamide gels).

Chem Phys Lipids, 1975 Sep, 15(1), 15 - 26
The activation of phospholipase C from Clostridium Welchii by quinine: an absolute requirement for calcium ions; Klein R et al.; Quinine activates the hydrolysis of phosphatidyl choline suspensions by phospholipase C (E.C . 3.1.4.3) obtained from Clostridium welchii . Low levels of calcium are an absolute requirement for this activation: Mg2+, Ba2+, Sr2+, and Zn2+ are ineffective . The induction period, or lag phase for this enzyme is dependent upon both calcium concentration and substrate interfacial surface area . At low concentrations (less then 50 muM) calcium ions affect the induction period but not the maximal rate of hydrolysis, whereas guinine predominantly affects the rate of hydrolysis by alterations in the surface charge carried by the substrate.

Infect Immun, 1975 Sep, 12(3), 536 - 43
Affinity chromatography purification of Clostridium perfringens enterotoxin; Scott VN et al.; Anti-enterotoxin immunoglobulins immobilized on CH-Sepharose or CNBr-Sepharose were used for affinity chromatography purification of Clostridium perfringens enterotoxin . Cell extracts containing enterotoxin or partially purified toxin preparations were applied to the column and nonspecifically-bound protein was eluted . NaOH was used to elute specifically bound toxin . The purity of enterotoxin purified by Sephadex G-100 chromatography followed by affinity chromatography appears similar to toxin highly purified by conventional means . The procedure can be used successfully for the rapid (less than 2 h) purification of small amounts of enterotoxin.

Ann Microbiol (Paris), 1975 Sep, 126(2), 175 - 80
{Comparative study of neuraminidases from "Diplococcus pneumoniae" and "Clostridium perfringens"}; Houdret N et al.; Neuraminidases have been purified from the culture medium of two microorganisms, one aerobic, Diplococcus neumoniae, the other anaerobic, Clostridium perfringens . The enzymatic properties of the 2 neuraminidases have been studied (pH optimum; effect of cations; activity toward different substrates: neuraminyllactose, dilactaminyllacto-N-tetraose, gangliosides, alpha1-acid glycoprotein, Collocalia glycoprotein, ovine submaxillary mucin, porcin intestinal and human bronchial mucins).

Arch Microbiol, 1975 Aug 28, 104(3), 237 - 40
The active species of "CO2" utilized in ferredoxin-linked carboxylation reactions; Thauer RK et al.; The active species of "CO2", i.e . CO2 or HCO-3(H2CO3) utilized by enzymes catalyzing ferredoxin-linked carboxylation reactions was determined . The enzyme investigated was pyruvate synthase from Clostridium pasteurianum (EC 1.2.7.1; Pyruvate: ferredoxin oxidoreductase) . Data were obtained which were compatible with those expected if CO2 is the active species . The dissociation constant (Ks) of the enzyme-CO2 complex was measured . At pH 7.2 Ks for CO2 of pyruvate synthase was found to be approximately 5 mM.

Eur J Biochem, 1975 Aug 15, 56(2), 427 - 32
On the biosynthesis of 5-methoxybenzimidazole . Precursor-function of 5-hydroxybenzimidazole, benzimidazole and riboflavin; Wurm R et al.; 1 . In Clostridium thermoaceticum 5-hydroxybenzimidazole is methylated to 5-methoxybenz-imidaxole and transformed to 5-methoxybenzimidazolylcobamide . 5-Hydroxybenzimidazolycobamide is also methylated to 5-methoxybenzimidazolylcobamide . These results indicate a possible precursor function of 5-hydroxybenzimidazole in the biosynthesis of 5-methoxybenzimidazole . 2 . The same microorganism uses benzimidazole to form benzimidazolylcobamide . This or externally added benzimidazolylcobamide, although taken up by the cells, is not further transformed (i.e . hydroxylated and methylated to 5-methoxybenzimdazolylcobamidel) . This excludes a precursor function of benzimidazole in the biosynthesis of 5-methoxybenzimidazole . 3 . Contrary to the biosynthesis of 5,6-dimethylbenzimidazole, 5-methoxybenzimidazole is not formed from riboflavin, but riboflavin inhibits the growth and the production of 5-methoxybenzimidazolylcobamide in Clostridium thermoaceticum . A tentative scheme for the biosynthesis of 5-methoxybenzimidazole via a riboflavin analog is discussed.

Biochemistry, 1975 Aug 12, 14(16), 3642 - 7
Replacement of acyl and alk-1-enyl groups in Clostridium butyricum phospholipids by exogenous fatty acids; Khuller GK et al.; The effect of exogenous unsaturated fatty acids on the acyl and alk-1-enyl group composition of the phospholipids of Clostridium butyricum has been examined . Unsaturated fatty acids support the growth of this organism in the absence of biotin . When cells were grown at 37 degrees in media containing oleate or linoleate and a Casamino acid mixture containing traces of biotin, the exogenous fatty acids were found mainly in the alk-1-enyl chains of the plasmalogens with less pronounced incorporation into the acyl chains . However, at 25 degrees in this medium, both the acyl and alk-1-enyl chains contained substantial amounts of the 18:1 supplement plus the C19-cyclopropane chains derived from it . Ak-1-enyl chains in all the major phosphatide classes showed a uniformly high substitution by the oleate supplement in cells grown at 37 degrees . The oleate and C19-cyclopropane content of the acyl chains was more variable among the phosphatide classes . At 37 degrees, trans-9-octadecenoic acid (elaidic acid) also supported growth and was incorporated into both acyl and alk-1-enyl chains at a high level . When cells were grown on oleate at 37 degrees in media containing biotin-free Casamino acids, both the acyl and alk-1-enyl chains had a high level of 18:1 plus C19-cyclopropane chains . In the cells grown at 37 degrees with oleate substantial changes were seen in the phospholipid class composition . There was a large decrease in the ethanolamine plus N-methylethanolamine plasmalogens with a corresponding increase in the glycerol acetals of these plasmalogens . The glycerol phosphoglycerides were also significantly lower with the appearance of an unknown, relatively nonpolar phospholipid fraction.

Biochem J, 1975 Aug, 149(2), 471 - 4
X-ray photoelectron spectra of iron-sulphur proteins; Andrews PT et al.; The X-ray photoelectron spectra of the 2p, 3s and 3p levels of iron in oxidized Clostridium pasteurianum ferredoxin indicate that the eight iron atoms in the molecule are indistinguishable . Their magnetic state is indicated both by core polarization splitting of the 3s electrons, and by "shake-up' satellites on the 2p lines . Similar satellites are observed in the 2p lines of reduced Chromatium high-potential iron-sulphur proteins and oxidized spinach ferredoxin, indicating that there too the iron atoms are magnetic . The low observed magnetic susceptibility of these proteins is therefore due to spin-coupling between the iron atoms in the active centre.

J Med Microbiol, 1975 Aug, 8(3), 429 - 35
The nuclear dehydrogenation of steroids by intestinal bacteria; Goddard P et al.; We have postulated that bacteria able to dehydrogenate the bile-acid nucleus are important in the aetiology of cancer of the colon . In this paper we report on screening for the ability to carry out two such reactions . The relevant enzymes are produced by a high proportion of strains of Clostridium paraputrificum, C . tertium and C . indolis, and by small numbers of strains in other clostridial species, but not by organisms of the other genera tested . Strains able to dehydrogenate the bile-acid nucleus represent a high proportion of the lecithinase-negative clostridia isolated from faeces of people living in Britain but a low proportion of those from people living in Uganda or Hong Kong.

Hoppe Seylers Z Physiol Chem, 1975 Aug, 356(8), 1203 - 8
{The occurrence of a reductase of delta2-carboxylic acids in Clostridium kluyveri with a stereospecificity different from that of butyryl-CoA dehydrogenase (author's transl)}; Hashimoto H et al.; A Clostridia strain (R-strain) which hydrogenates tiglinate (1b) and alpha-methylcinnamate (1c) in the presence of hydrogenase gas in 2H2O to (2R, 3S)2-methyl-{2,3-2H}butyrate (5b, H = 2H) and (alphaR, betaR)alpha-methyl{alpha,beta-2H}dihydrocinnamate (5c, H = 2H), respectively, was isolated . The configuration at C-3 was determined by 1H-NMR spectroscopy in the presence of Eu(fod)3 . The stereochemistry of this hydrogenation is the mirror image of that which has been determined with intact cells of another strain of Clostridium kluyveri (S-strain) . In the presence of hydrogen gas, the R-strain hydrogenates crotonate in 2H2O to butyrate with the following deuterium distribution: C-2, 1.85; C-3, 1.35; and C-4, 0.63 deuterium atoms . Crotonate seems to be the substrate of two reductases with sterically different actions . Tiglinate (1b) and alpha-methylcinnamate, however, are hydrogenated only by that reductase which is different from the butyryl-CoA dehydrogenase.

Hoppe Seylers Z Physiol Chem, 1975 Aug, 356(8), 1195 - 201
{On the specifity and stereospecificity of the conversion of different 2,3-unsaturated acids by Clostridium kluyveri (author's transl)}; Hashimoto H et al.; Further 2,3-unsaturated acids are revealed which can be reduced by Clostridium kluyveri with crotonate or butyrate as hydrogen donors . Unsaturated and saturated 3-halogenated acids are transformed into the saturated halogen-free acids . The following reaction sequence is proposed: a) hydrogenation, b) elumination of HX and c) again hydrogenation . Tiglinate ((E)-2-methyl-2-butenoate) and (E)-2-methyl-2-pentenoate are stereospecifically reduced to the (S)-2-methyl substituted acids . C . kluyveri contains endogenous material; in the presence of hydrogen acceptors such as 2,3-unsaturated acids this is degraded to acetate, and the reducing equivalents liberated hydrogenate the unsaturated acid . In a transient phase the hydration products of the unsaturated acids are present in the non-activated form in appreciable amounts . Tiglinate as well as crotonate is partially converted to ethyl methyl ketone and aceton and/or propanol, respectively.

Proc Natl Acad Sci U S A, 1975 Aug, 72(8), 2868 - 72
Synthetic analogs of active sites of iron-sulfur proteins: bis (o-xylyldithiolato) ferrate (III) monoanion, a structurally unconstrained model for the rubredoxin Fe-S4 unit; Lane RW et al.; To complete the set of synthetic analogs of the three recognized types of active sites in iron-sulfur redox proteins, the compound (Et4N){Fe((SCH2)2C6H4)2}, derived from o-xylyl-alpha,alpha'-dithiol, has been prepared and its structure has been determined by x-ray diffraction . The bischelate anion contains a near-tetrahedral Fe(III)-S4 coordination unit with small rhombic distortions and all Fe-S bond distances in the range 2.252-2.279 A . Its electronic properties have been partially characterized by measurement of electronic absorption, paramagnetic resonance, Mossbauer spectra, and magnetic susceptibility . The analog, as the protein, exhibits the Fe(III)/Fe(II) redox couple . These results substantiate designation of {Fe((SCH2)2C6H4)2}- as a synthetic analog of the Fe(III)(S-Cys)4 center in oxidized rubredoxin proteins . Comparison of the analog structure with that of the Clostridium pasteurianum rubredoxin active site shows that the former is substantially less distorted from idealized tetrahedral symmetry, and is considered to represent an essentially unconstrained structural model of the latter . Provided the grossly distorted tetrahedral stereochemistry of the protein site persists through final structural refinement, the analog-protein structural comparison supports an entatic state description of oxidized rubredoxin.

J Gen Microbiol, 1975 Aug, 89(2), 337 - 44
The effect of chlorine on spores of Clostridium bifermentans, Bacillus subtilis and Bacillus cereus; Wyatt LR et al.; The effect of chlorine on the germination, outgrowth, colony formation and structure of spores of Clostridium bifermentans, Bacillus subtilis var . niger and Bacillus cereus was examined . Chlorine decreased heat resistance and slowed or prevented germination and swelling, but spores that did swell were usualy able to elongate to form vegetative cells . Chlorine removed protein from spores, apparently from the coat, and allowed lysozyme to initiate germination . Treatment with other agents that remove spore-coat protein increased the lethal effect of chlorine by as much as 4000-fold, suggesting that coat protein protects spores against chlorine.

Can J Microbiol, 1975 Aug, 21(8), 1259 - 69
{Nitrosoguanidine-induced attenuated Clostridium perfringens type A mutant in gas gangrene}; Lapointe JR et al.; A fully virulent classical type A strain of Clostridium perfringens was treated during its logarithmic growth phase with 100 mug/ml of N-methyl-N'-nitro-N-nitrosoguanidine, the bacteria being exposed to the mutagen for 30 min at 37 degrees C in a phosphate buffer adjusted to pH 6.2; after treatment the suspension was streaked on sheep blood agar plates, and colonies that showed an alteration in the theta-hemolysis pattern were selected for isolation . The virulence of two mutants, thus altered in their theta-hemolysis, was studied . One, designated LNG 5, was still capable of killing most of the inoculated guinea pigs in less than 24 h with all the clinical, macroscopic, and bacteriological signs of gas gangrene; however, histological sections showed that tissue damage was not as marked as with the wild strain . On the contrary, the second mutant, labelled LNG 11, was completely avirulent as far as gas gangrene was concerned; indeed, the injection of fluid cultures containing 1 times 10(8) - 10(9)/ml viable bacteria, was not followed by any clinical, bacteriological, or histological signs of gas gangrene . However, strain LNG 11 did give rise to a firm swelling of the inoculated thigh with a corresponding acute inflammatory response of the connective tissue, although the muscle fiber was unaltered . Eventually, this local reaction was followed by necrosis of the skin accompanied by an acute or subacute inflammation with fibroblastic proliferation . These superficial lesions healed spontaneously . They could not be reproduced with crude filtrate alone or with washed bacilli . Strain LNG 11 was therefore considered to be soletly an attenuated strain since, although avirulent as far as gas gangrene is concerned . it is still capable of producing low levels of toxic material . This appears to be the first time that such a strain of C . perfringens type A has been obtained by nitrosoguanidine treatment.

Appl Microbiol, 1975 Aug, 30(2), 319 - 23
Inhibition of Clostridium botulinum by strains of Clostridium perfringens isolated from soil; Smith LD; Thirty-one soil samples were examined for the presence of organisms capable of inhibiting growth and toxin production of strains of Clostridium botulinum type A . Such organisms were found in eight samples of soil . Inhibiting strains of C . perfringens were found in five samples, of C . sporogenes in three and of Bacillus cereus in three . Three of the C . perfringens strains produced an inhibitor effective on all 11 strains of C . botulinum type A against which they were tested, seven of eight proteolytic type B strains, one nonproteolytic type B strain, five of nine type E strains and all seven type F strains, whether proteolytic or nonproteolytic . They did not inhibit any of 26 type C strains, 6 type D strains, 4 type E strains, or 24 C . sporogenes strains . In mixed culture, an inhibitor strain of C . perfringens repressed growth and toxin production by a C . botulinum type A strain even though it was outnumbered by the latter about 40 times . It also repressed growth and toxin production of C . botulinum in mixed culture of soils in which this latter organism naturally occurred when cooked meat medium but not when trypticase medium was used.

J Bacteriol, 1975 Aug, 123(2), 419 - 27
Isolation and characterization of Clostridium perfringens mutants altered in both hemagglutinin and sialidase production; Rood JI et al.; The relationship between the production of hemagglutinin and sialidase activities by Clostridium perfringens was investigated by screening for mutants producing reduced levels of hemagglutinin activity . Twelve mutants were isolated; all produced reduced levels of sialidase activity and several had other altered phenotypic markers . Revertants that regained the ability to produce active hemagglutinin were isolated . All of these revertants produced increased sialidase activity . These results show that the production of hemagglutinin activity is directly related to the production of sialidase activity . Evidence is also presented that the processes of sporulation and the production of extracellular proteins are interrelated.

Infect Immun, 1975 Aug, 12(2), 419 - 32
Rabbit corneal damage produced by Pseudomonas aeruginosa infection; Gray LD et al.; Gross, light microscopic, and electron microscopic examination of the rabbit corneal destruction produced by experimental Pseudomonas aeruginosa infections revealed a combination of acute inflammation and liquefaction necrosis of the cornea . Degeneration of the epithelial cells and the start of polymorphonuclear leukocyte infiltration of the cornea occurred initially . These changes were followed by loss of the epithelium, degeneration and loss of the keratocytes and endothelium, loss of the characteristic weblike pattern of the proteoglycan ground substance, dispersal of ultrastructurally normal collagen fibrils, extensive accumulation followed by degeneration of polymorphonuclear leukocytes, and accumulation of plasma proteins and fibrin in the necrotic cornea . Histochemical examination of the cornea suggested a loss of the proteoglycan ground substance but not of collagen . Rabbit corneas injected with Clostridium histolyticum collagenase showed gross and cellular changes similar to those observed during the pseudomonal infections; however, histochemical examination suggested a loss of collagen, and electron microscopy revealed ultrastructurally abnormal collagen fibrils . The results support the idea (i) that a bacterial or host-derived collagenase is not required for extensive corneal damage during a P . aeruginosa corneal infection, and (ii) that a P . aeruginosa corneal infection may severly damage the cornea by producing extensive corneal edema and by causing the loss of the corneal proteoglycan ground substance, thus resulting in dispersal of undamaged collagen fibrils, weakening of the cornea, and subsequent descemetocele formation and corneal perforation by the anterior chamber pressure.

Infect Immun, 1975 Aug, 12(2), 225 - 32
Sensitive assay for detection of toxin-induced damage to the cytoplasmic membrane of human diploid fibroblasts; Thelestam M et al.; A sensitive assay was developed for detection and quantitation of subtle permeability changes in the cytoplasmic membrane of human diploid fibroblasts . Release of the non-metabolizable amino acid {1-14C}alpha-aminoisobutyric acid (AIB; molecular weight (103) from the cytoplasm of prelabeled cells was used as an indicator of toxin-induced membrane damage . An optimal procedure for labeling these cells was designed after varying the conditions with regard to pH, temperature, concentration of AIB, composition of medium, and incubation time . Toxin-induced release of AIB was compared with release of a previously described nucleotide label, {3H}uridine . Melittin from bee venom and the polyene antibiotics filipin and amphotericin B in low concentrations induced a strikingly greater release of AIB than of nucleotide label . The sensitivity of this assay was furthermore demonstrated by treatment with the following bacterial cytolysins: phospholipase C and theta-toxin from Clostridium perfringens, alpha-, beta-, delta-, and gamma-toxins from Staphylococcus aureus, and streptolysin S from Streptococcus pyogenes . In spite of their different modes of action, all these membrane-active toxins at low concentrations induced a significant release of AIB label . For an equal release of nucleotide label, several times higher concentrations were required.

Infect Immun, 1975 Aug, 12(2), 440 - 2
Measurement of biological activities of purified and crude enterotoxin of Clostridium perfringens; Nilo L; Enterotoxin of Clostridium perfringens was assayed and compared with toxicity in mice and erythemal activity in guinea pigs . Conversion factors were used to express these biological activities of crude enterotoxin in terms of weight of pure enterotoxin protein . One microgram of enterotoxin was equivalent to 3.41 erythema units and to 0.68 mouse median lethal dose.

Infect Immun, 1975 Aug, 12(2), 274 - 80
Effects of Clostridium perfringens enterotoxin on metabolic indexes of everted rat ileal sacs; McDonel JL et al.; Everted sacs of rat ileum were incubated in Ringer phosphate solution while oxygen uptake, glucose uptake, and lactate production were determined . Sacs treated with Clostridium perfringens enterotoxin, in the form of crude cell-free extract and purified protein, consumed significantly less oxygen than untreated sacs . However, the toxin-treated sacs took up glucose and produced lactate at levels that were not significantly different than those observed in controls . We conclude that oxidative metabolism is inhibited by the action of the toxin, whereas conversion of glucose to lactate via glycolysis seems unaffected

C R Acad Sci Hebd Seances Acad Sci D, 1975 Jul 28, 281(4), 317 - 9
{Transfer of the tetracycline-chloramphenicol plasmid in Clostridium perfringens}; Sebald M et al.; Two plasmids which confer resistance to Tetracycline-Chloramphenicol and Erythromycine-Clindamycine were found in a strain of Clostridium perfringens . The plasmid (Tet-Chl) was shown to be transferable to sensitive strains of C . perfringens . The transfer experiments were made with the wild type strain or strains cured by one plasmid or the other as donor strains.

N Z Med J, 1975 Jul 23, 82(544), 52 - 4
Food poisoning--four unusual episodes; Begg RC; Four unusual outbreaks of food poisoning occurring in the Dunedin Health District during the period 1971-1973 are described . These involved a contaminated cordial, a death associated with a Clostridium perfringens outbreak, salmonellosis and infectious hepatitis in persons eating uncooked shellfish and symptoms associated with the ingestion of a normally edible fish--the trumpeter.

Biochim Biophys Acta, 1975 Jul 21, 400(1), 162 - 6
The application of collagenase, purified by affintiy chromatography, to the isolation of insoluble elastin from bovine aorta; Spina M et al.; Clostridium histolyticum collagenase (clostridiopeptidase A, EC 3.4.4.16), purified by affinity chromatography, was applied to the isolation of insoluble elastin from bovine aorta . The extremely low level of N-terminal residues (1.6 mol per 10(6) g of protein) present in this preparation indicated the almost complete lack of hydrolytic damage caused by the isolation procedure . The amino acid profile of the aortic elastin was found to be almost identical to that of insoluble elastin prepared from bovine ligamentum nuchae by the same method.

Zentralbl Bakteriol {Orig A}, 1975 Jul, 232(2-3), 303 - 7
{Serological examinations with apathogenic clostridia (M 55) (author's transl)}; Gericke D et al.; Spores of the apathogenic clostridium butyricum (M 55) germinate after intravenous application in mammalians in tumor tissue only . On the basis of these earlier findings a diagnostic and prognostic procedure, used so far, namely the agglutinationtest with vegatative forms of this clostridium was improved by creation of a hemagglutination reaction . With different methods some different hemagglutinins have been isolated . Their efficacy was tested against specific antibodies in rabbits and mice . The higher sensitivity of these procedures was demonstrated.

Infect Immun, 1975 Jul, 12(1), 143 - 7
Rabbit immunoglobulin responses to the flagella, somatic, and protective antigens of a highly protective strain of Clostridium chauvoei; Chandler HM; The immunoglobulin response of rabbits to the flagells (H), somatic (O), and protective antigens of a highly protective strain of Clostridium chauvoei was studied using antisera that had been fractionated by Sephadex G-200 chromatography . The H antigen elicited the characteristic agglutinin response to a protein antigen--early production of 19S globulin followed by persistent 7S globulin production . The O antigen stimulated a transient agglutinin response which was detected in both the 19S and 7S serum fractions . Protective antibody was assayed by passive protection tests in mice . Using these tests the protective activity of the rabbit sera was found to be confined exclusively to the 7S serum fractions . Purified immunoglobulin G, prepared by DEAE-cellulose chromatography of the above sera, was also tested and found to confer considerable passive protection on mice . It is considered that either the protective antigen fails to stimulate an immunoglobulin M response or that immunoglobulin M is relatively ineffective in conferring protection against infection in the mouse passive protection tests.

Can J Surg, 1975 Jul, 18(4), 339 - 41, 344
Fulminating nonclostridial gas-forming infection: a case of necrotizing fasciitis; Blanchard RJ; Perianal infection in a 40-year-old man resulted in extensive necrotizing fasciitis of the retroperitoneal space and septic shock . Despite the fact that radiography revealed linear streaking in the belly of the psoas muscle due to gas formation, the nature of the infection was necrotizing fasciitis and not myonecrosis . This contradicts Brightmore's contention that such a finding always indicates clostridial myonecrosis . Despite the absence of Clostridium welchii, necrotizing fascitis is none the less extremely serious, usually occurring in the limbs or abdominal wall superficial to muscle layers . The case reported is unusual in that infection affected the fascia deep to abdominal muscles in the retroperitoneal space, where surgical exposure is difficult . An appropriate surgical approach afforded adequate treatment.

Can J Microbiol, 1975 Jul, 21(7), 1136 - 8
Characterization of two reserve glucans from Clostridium pasteurianum; Brown RG et al.; The reserve glucans of C . pasteurianum, previously assumed to be a single polysaccharide, have been fractionated into two polysaccharides which resemble amylopectin and dextran . Both polysaccharides are produced when cells are grown with sucrose, D-glucose, or D-fructose as carbon sources.

J Virol, 1975 Jul, 16(1), 107 - 15
Development of bacteriophage F1 in Clostridium sporogenes: characterization of RNA transcripts; Taylor DE et al.; RNA transcription was investigated during the development of F1 phage, which is specific for the strict anaerobe Clostridium sporogenes . RNA species transcribed during F1 phage infection were characterized with respect to time of appearance and molecular weight by polyacrylamide gel electrophoresis . Ten mRNA species were characterized, of which five were produced early in infection and five were synthesized late in infection . All the above 10 species were transcribed from one strand of F1DNA, the heavy strand . Two additional mRNA species were transcribed from the light strand of F1 phage DNA later in infection . Throughout the F1 phage infective cycle, rRNA was continuously synthesized by cells of C . sporogenes.

Surg Gynecol Obstet, 1975 Jul, 141(1), 35 - 9
Streptococcal cellulitis of the scrotum and penis with secondary skin gangrene; Haury B et al.; Cellulitis of the scrotum and penis is caused, in the majority of instances, by a beta hemolytic streptococci without a discernible portal of entry . Clostridium, occasionally, will result in this disease as a manifestation of a perirectal abscess . In either instance, fluid accumulates rapidly in the closed space between Colles' and Buck's fascia, producing intense swelling of the scrotum . If this compartment is not immediately decompressed by linear incisions, devascularization of the scrotal and penile skin will often occur, resulting in gangrene . Immediate treatment of the bacterial infection with penicillin also is essential . If gangrene does develop, radical debridement of the necrotic tissue as well as a wide margin of adjacent inflamed skin must be undertaken . Continual monitoring of the microflora of the debrided would is essential for the selection of the appropriate antibiotic against any secondary intruders . Coverage of the granulating would is accomplished when the would bacterial count is below 10-5 per gram of tissue.

Can J Surg, 1975 Jul, 18(4), 372 - 5, 378
Bacterial peritonitis and the bursting strength of intestinal anastomoses; Kilam SK et al.; In order to study the bursting strength of intestinal anastomoses, resection of a wedge of ileum and creation of an anastomosis were performed, and peritonitis was induced in five groups of rats . Peritonitis was induced by intraperitoneal injection of bacterial suspensions of Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Clostridium welchii, and the exteriorized ileal loop in each rat was regularly inspected for peritoneal exudation, adhesions, hyperemia, obstruction, leakage and bursting strength . All bowel anastomoses performed in the presence of peritonitis were weaker than those done in a sterile field (control group) and the type of organism influenced the strength of the anastomosis and the histopathologic changes; in this study the weakest anastomoses were associated with E . coli infection.

Zentralbl Bakteriol {Orig B}, 1975 Jul, 160(4-5), 432 - 42
{A comparison of the assay regulations for sterility testing between the USP XIX and the European Pharmacopoeia (author's transl)}; Seyfarth H; The USP XIX is to be published in the summer 1975 . In this paper the most important passages of its sterility test are compared with the regulations of the EP 1 . So a test should be rendered possible to come up to both Pharmacopoeia . a) Both the membrane filtration method and the direct inoculation of media may be used as the test methods . The membrane filtration method is the method of choice . But the mentioned pore diameter of 0.45 +/- 0.02 mum is too large . Those filters with a pore diameter of 0.2 +/- 0.02 mum are preferable . b) While EP 1 does not mention the culture media, USP XIX prescribes Fluid Thioglycollate Medium and Soybean-Casein Digest Medium . These culture media are not sufficient . The use of Fluid Thioglycollate Medium, Soybean-Casein Digest Medium and Fluid Sabouraud Medium is suggested . c) The same organisms are recommended for the control of the culture medium as well as for the determination of the minimal inhibiton concentration (Clostridium sporogenes, Bacillus subtilis, Staphylococcus aureus, Candida albicans and perhaps a mould fungus) . d) In the USP the number of samples is decided by the risk of contamination . This is preferable to the EP 1, where the number of samples is fixed by the size of the charge . e) While EP 1 lays down 7 days as an incubation time for bacteria as well as fungi, the USP XIX prescribes a time between 7 and 14 days, depending on the contamination risk and on the method used . As a compromise 10 days for bacteria and 14 days for fungi are recommended.

Lab Anim, 1975 Jul, 9(3), 233 - 9
A review of 105 necropsies in captive baboons (Papio cynocephalus); Kim JC et al.; In contrast to the findings in newly caught babboons in Africa, the leading cause of death in captivity in America was a pneumonia and enteritis complex . Bacterial species such as Proteus morgani, Proteus mirabilis, Clostridium sp., Pseudomonas paracolon, pathogenic Escherichia coli and cocci have been isolated in association with a pneumoenteric syndrome of 35 newborn baboons, including neonates . The majority of these animals died with pulmonary factors such as anoxia and pneumonia.

Zentralbl Bakteriol {Orig A}, 1975 Jul, 232(2-3), 294 - 302
{Toxigenesis of Clostridium novyi type A . 2 . Communication: Extraction alpha-toxin from the cells by hypertonic buffers (author's transl)}; Schallehn G; Great amounts of alpha-toxin produced by Cl . novyi type A and accumulated within the cell can be extracted by 0,2M sodium citrate buffer (pH 7,7) during the course of 2 to 5 days at 37 degrees C . Physiological saline, 0,1 M sodium citrate buffer (pH 7,7) and 0,05 to 0,2 M EDTA-buffers (pH 7,7) were not effective for complete extraction . During the process of extraction the lattice like structure of the peripheral part of the Cl . novyi cell wall was partially destroyed . The lattice is supposed to be a diffusion barrier for the toxin and may be regarded as one reason for the toxin accumulation in Cl . novyi.

Eur J Biochem, 1975 Jul 1, 55(2), 455 - 63
Heterogeneity of enterotoxin-like protein extracted from spores fo Clostridium perfringens type A; Frieben WR et al.; Enterotoxin-like protein was extracted from spores of three enterotoxin-positive and three enterotoxin-negative strains of Clostridium perfringens type A by urea/mercaptoethanol, alkaline mercaptoethanol and alkaline dithiothreitol . Disc immunoelectrophoresis demonstrated that three distinct enterotoxin-like proteins could be extracted . In 7% acrylamide gels, type I, type II, and type III enterotoxinlike proteins had relative mobilities of 0.52, 0.63, and 0.73 respectively . In contrast to disc immunoelectrophoresis, immunoelectrophoresis in agar gel demonstrated identical electrophoretic properties for the various entertoxin-like proteins . Immunoelectrofocusing experiments gave isoelectric points of 4.43, 4.43, 4.36, and 4.52 for purified entertoxin and type I, type II, and type III enterotoxin-like proteins respectively . Ferguson plots (i.e., log relative mobility versus acrylamide concentration) yielded nonparallel lines which intersected at a nonsieving concentration of acrylamide indicating that the various species of enterotoxin-like protein differed in size . Estimation of the molecular weight of purified enterotoxin and the three species of enterotoxin-like protein was done by comparing the slopes obtained in Ferguson plots with those obtained using proteins of a known molecular weight . Molecular weights of 38000, 36500, 23000, and 15400 were obtained for purified enterotoxin, type I, type II, and type III enterotoxin-like protein respectively . Collectively, the evidence indicates that fractionation of the different species of enterotoxin-like protein was due primarily to differences in their size, and that different forms of enterotoxin-like protein can be extracted from spores of different strains of C . perfringens type A.

Br J Surg, 1975 Jul, 62(7), 518 - 9
Fatal Clostridium welchii septicaemia following acute cholecystitis; Clancy MT et al.; A case of Clostridium welchii septicaemia following acute cholecystitis is described . The onset was acute and a rapidly fatal outcome ensued . Radiological findings were negative . An approach to the antibiotic treatment and general management is discussed.

Poult Sci, 1975 Jul, 54(4), 1000 - 7
Reduction of Clostridium perfringens by feed additive antibiotics in the ceca of chickens infected with Eimeria tenella; Arakawa A et al.; Two experiments were performed ot investigate the effect of feed additive antibiotics on Clostridium perfringens and Enterobacteriaceae in the ceca of chickens infected with Eimeria tenella . In the first experiment, chickens were continuously fed rations containing thiopeptin, 2 mg./kg.; bacitracin, 20 mg./kg.; penicillin, 12 mg./kg.; or chlortetracycline, 22 mg./kg . One day after antibiotic feed was given, each bird received an oral inoculation of 30,000 E . tenella oocysts . The growth of C . perfringens was stimulated by the infection in unmedicated chickens . Dietary thiopeptin, bacitracin, penicillin, or chlortetracycline suppressed the number of C . perfringens recovered 5 and 7 days after infection . Enterobacteriaceae were increased by the infection, but dietary antibiotics did not reduce the increase . In the second experiment, chickens were given feed containing amprolium plus ethopabate, 125 plus 8 mg./kg., and a combination of the coccidiostat and one of 4 antibiotics: thiopeptin, bacitracin, penicillin, or chlortetracycline . Birds were each given an oral inoculation of 30,000 coccidiostat-resistant E . tenella oocysts . Infection resulted in an increase of C . perfringens in the unmedicated control and the coccidiostat-treated groups . Dietary thiopeptin, bacitracin, penicillin, or chlortetracycline reduced the number of C . perfringens found 5 and 7 days after infection . Counts of Enterobacteriaceae were increased by the infection, but dietary antibiotics did not suppress the increased counts . In both experiments, dietary administration of antibiotics did not reduce gross cecal lesions.

Eur J Biochem, 1975 Jul 1, 55(2), 445 - 53
The internal-alkaline pH gradient, sensitive to uncoupler and ATPase inhibitor, in growing Clostridium pasteurianum; Riebeling V et al.; 1 . The intracellular pH was measured in growing Clostridium pasteurianum with and acid-base equilibrium distribution method . {14C}Dimethyloxazolidinedione, {14}methylamine and {14C}acetic acid were used as "deltapH-indicators" . During growth the extracellular pH decreased from 7.1 to 5.1; simultaneously the intracellular pH changed from 7.5 to 5.9 . Thus, the intracellular pH was more alkaline than the extracellular pH by 0.4 to 0.8 pH-units . 2 . This pH gradient (interior alkaline) was abolished by the proton conductor carbonylcyanide m-chlorophenylhydrazone and the ATPase inhibitor N,N'-dicyclohexylcarbodiimide . The pH gradient could not be demonstrated in cells depleted of an energy substrate . These results suggest that the pH gradient is formed by an ATPase-driven extrusion of protons from the cells rather than by a Donnan potential . 3 . Growth of the organism was inhibited by low concentrations of both carbonylcyanide m-chlorophenylhydrazone (5 muM) and dicyclohexylcarbodiimide (5 muM) . This finding suggests that the pH gradient is essential for the growing cell as it may be required for substrate accumulation and other types of transport processes.

Eur J Biochem, 1975 Jun 16, 55(1), 111 - 7
The active species of 'CO2' utilized by reduced ferredoxin:CO2 oxidoreductase from Clostridium pasteurianum; Thauer RK et al.; Reduced ferredoxin:CO2 oxidoreductase (CO2 reductase) from Clostridium pasteurianum catalyzes the reduction of 'CO2' to formate with reduced ferredoxin, an isotopic exchange between 'CO2' and formate in the absence of ferredoxin, and the oxidation of formate to 'CO2' with oxidized ferredoxin . The active species of 'CO2', i.e . CO2 or HCO3 (H2CO3), utilized by the enzyme was determined . The method employed for the species identification was that of Copper et al . (1968) . Both 'CO2' reduction to formate and the exchange reaction were studied . Data were obtained which are compatible with those expected if CO2 is the active species . The V and the dissociation constant Ks of the enzyme - CO2 complex in dependence of pH were determined from initial velocity studies of the exchange reaction . V was found to be only slightly affected by pH between 5.5 and 7.5 . Ks was markedly dependent on pH; the constant increased with decreasing pH from 0.2 mM at pH 7.5 to 3 mM at pH 5.5.

Zentralbl Bakteriol {Orig A}, 1975 Jun, 232(1), 91 - 9
{Toxigenesis of Clostridium onvyi type A . 1 . Communication: production of alpha-toxin in vitro (author's transl)}; Schallehn G; The production and excretion of alpha-toxin in vitro was studied with two pathogenetically different strains of Cl . novyi type A . During the exponential phase of growth nearly all alpha-toxin produced by strain 955 was concentrated within the cell, while only small amounts of toxin could be detected in the culture medium . On the contrary, alpha-toxin produced by strain Fo during the exponential phase of growth was found in equal parts (240 DLM/2 times 10(8) cells) within and outside the cell . During the stationary phase of growth 50 to 75% of the intracellular alpha-toxin was released into the culture fluid . The different rates of excretion of alpha-toxin may be the reason for the differing phenomena in gas gangrene of guinea pigs infected with Cl . novyi.

Zentralbl Bakteriol {Orig A}, 1975 Jun, 232(1), 100 - 4
{Clostridium novyi type A bacteriophages (author's transl)}; Schallehn G et al.; In the culture fluids and on the cells of two toxigenic strains of Clostridium novyi type A, bacteriophages were detected and confirmed by electron microscopy (Fig . 1 and 2) . The heads of the bacteriophages are hexagonal and 500 A in diameter . The tails have a length of 1400 A and a diameter of 60 A . Reproduction of the bacteriophages was not yet possible owing to the lack of sensitive indicator strains.

J Trauma, 1975 Jun, 15(6), 515 - 8
Clostridium sordelli infection; Browdie DA et al.; A case of human Clostridium sordelli soft tissue infection is presented . Analysis of this patient's course led to the use of a mouse experimental model for examination of this organism's potential for toxin production . Data thus obtained correlated with that seen in this instance of human infection, indicates that the lethal effects of this organism may be related to the ability to Clostridium sordelli to produce a widespread "toxin-mediated" edema with subsequent marked "third-space" sequestration of fluid.

Jpn J Med Sci Biol, 1975 Jun, 28(3), 157 - 64
Responses of Clostridium botulinum type B and E progenitor toxins to some clostridial sulfhydryl-dependent proteases; Oishi I et al.; Sulfhydryl-dependent proteases produced by Clostridium botulinum types A, B, and F, Clostridium histolyticum, Clostridium sporogenes and Clostridium perfringens activate preferentially type E over type B progenitor toxin but less efficiently than trypsin . The results explain why activable toxin is demonstrable in culture of a strongly proteolytic type B strain.

Jpn J Microbiol, 1975 Jun, 19(3), 167 - 72
Observations on nonconverting phage, c-n71, obtained from a nontoxigenic strain of Clostridium botulinum type C; Oguma K et al.; A nontoxigenic mutant (C-N71) obtained from a toxigenic strain of Clostridium botulinum type C, Stockholm, with nitrosoguanidine treatment was found to be lysogenic by the lysis test . Although the filtrate of a passaged lysate of this nontoxigenic but lysogenic strain, C-N71, lysed cells of the nontoxigenic strain C-AO2 equally as well as the converting phage c-st obtained from the strain C-Stockholm, it did not convert C-AO2 to the toxigenic state . The lysis spectrum of this filtrate was the same as that of the c-st phage . The ability of the filtrate to lyse the indicator cells, C-AO2, was destroyed neither by trypsin nor DNase but was inactivated by heat treatment at 80 C for 10 min . This suggested that the agent which caused lysis was not boticin but probably a phage . An electron micrograph of the complete phage, c-n71, which was similar in morphology to that of the c-st phage was obtained from the filtrate of strain C-N71 . Anti-c-n71 phage rabbit serum neutralized both the lytic and the converting activities of the c-st phage . These findings strongly suggest that the c-n71 phage is a mutant of the c-st phage which lacks the gene controlling production of botulinum type C toxin.

Can J Microbiol, 1975 Jun, 21(6), 920 - 6
Observations on the distribution and ecology of Clostridium botulinum type E in Alaska; Miller LG; Environmental samples collected along the coastline and from the interior of Alaska were examined for the presence of Clostridium botulinum . Clostridium botulinum type E was detected in soils from 5 of 12 beaches; in 7 of 115 non-coastal soil samples; in sediments from six of eight locales; in gills of salmon from two fishing areas; and in the feces of 1 of 44 colonic samples from marine mammals . The basic biochemical characteristics of the isolates were determined . Tube tests for demonstrating gelatin liquefaction proved insensitive with these strains, whereas a plate test detected gelatinase in all isolates . The presence of multiple nidi and the continual discharge of organic materials into the environment may contribute to the perpetuation of botulinum spores by which foods prepared form marine animals become contaminated . An emphasis should be placed upon the need for measures to reduce environmental contamination, to reduce contamination during food preparation, and to alert continually the population of the hazard wherever botulism is endemic.

Appl Microbiol, 1975 Jun, 29(6), 802 - 6
Microflora of maize prepared as tortillas; Capparelli E et al.; Very little is known of the microflora in tortillas, the major component in the diet of many Guatemalans and other Central Americans . Based in a Guatemalan highland Indian village, this study examined the types and amounts of bacteria, yeasts, and molds in tortillas and in their maize precursors . Coliforms, Bacillus cereus, two species of Staphylococcus, and many types of yeast were the main contaminants, but low concentrations of alpha-hemolytic Streptococcus, facultative Clostridium, and other bacterial types were also found . When tortillas were first cooked, the bacterial counts dropped to 1,000 or fewer organisms per g, a safe level for consumption . Under village conditions, bacterial counts regained precooking levels (about 10(8) organisms/g) within 24 h and rose even higher after 48 h . Reheating caused very little change; hence, bacterial levels remained very high in old tortillas kept for later consumption . A search for the sources of contamination showed that most came from water used in preparation and from the soiled hands of women preparing the tortillas . As an attempt to correct certain nutritional needs of the population, the Institute of Nutrition for Central America and Panama initiated a tortilla fortification project in the Guatemalan village . The bacterial counts in fortified tortillas did not differ significantly from those in oridinary tortillas . Furthermore, the level of contamination was constant among tortillas of different sizes and among tortillas made from different types of maize.

J Biochem (Tokyo), 1975 Jun, 77(6), 1165 - 9
Origin and early evolution of transition element enzymes; Egami F; In this paper we speculate on the origin and early evolution of transition element enzymes . Iron, molybdenum, and zinc, the most abundant transition elements in seawater, presumably complexed with compounds accumulated in the primeval sea in the course of chemical evolution forming compounds with subsequently evolved to form proenzymes or early enzymes with low activity and broad specificity . Iron complexes may be regarded as precursors of electron transfer enzymes, molybdenum complexes as precursors of enzymes involved in the metabolism of small molecules, and zinc complexes as precursors of hydrolytic and transferring enzymes, including enzymes participating in the metabolism of macromolecules and information transfer . The different iron, molybdenum, and zinc enzymes found in bacteria including Clostridium may then have arisen through specialization by increases in the enzyme specificity of these proenzymes . Copper would have been incorporated as an enzyme constituent after the elevation of environmental redox potential, probably due to the accumulation of atmospheric oxygen.

Hoppe Seylers Z Physiol Chem, 1975 Jun, 356(6), 653 - 62
Reduced ferredoxin: CO2 oxidoreductase from Clostridium pasteurianum . Effect of ligands to transition metals on the activity and the stability of the enzyme; Thauer RK et al.; Reduced ferredoxin: CO2 oxidoreductase (CO2-reductase) from Clostridium pasteurianum catalyzes the reduction of CO2 to formate at the expense of reduced ferredoxin, an isotopic exchange between CO2 and formate in the absence of ferredoxin, and the oxidation of formate to CO2 with oxidized ferredoxin . The three activities were found to be equally affected by monovalent anions known to be ligands to transition metals: The enzyme was reversibly inhibited by azide (Ki = 0.004mM), cyanate (Ki = 0.3 mM), thiocyanate (Ki = 1mM), nitrite (Ki = 0.4mM), nitrate (Ki = 6mM), chlorate (Ki = 3mM), fluoride (Ki = 5mM), and by chloride, bromide, iodide (Ki greater than 5mM) . There was no observable effect of pH on the inhibition constants . The enzyme was not inhibited by carbon monoxide . The enzyme was irreversibly inactivated by low concentrations (10muM) of cyanide . The rate of inactivation increased with increasing pH with an inflection point near pH 9.5 . Reduced ferredoxin and formate rather than oxidized ferredoxin or CO2 protected the enzyme from inactivation by cyanide . The enzyme was protected by azide and cyanate from inactivation . In the presence of high concentrations of the monovalent anions the rate of inactivation by heat (55 degrees C), by molecular oxygen, and by cyanide was decreased by a factor of more than 100 . Half maximal protection was observed at the Ki concentrations of the two reversible inhibitors . The data are interpreted to indicate that a transition metal of weak "a class" character and a disulfide are catalytically significant groups of CO2-reductase from C . pasteurianum.

Hoppe Seylers Z Physiol Chem, 1975 Jun, 356(6), 1027 - 42
Purification and characterization of neuraminidase from Clostridium perfringens; Nees S et al.; Clostridium perfringens cells were cultivated on a large scale using an automatic system . Neuraminidase secreted by the cells into the culture medium was purified 380 000-fold by: precipitation with ammonium sulfate between 50 and 85% saturation, filtration on Sephadex G-75, electrophoresis on polyacrylamide gel, and by isoelectric focusing . Three enzyme fractions with different migration rates were obtained by preparative disc electrophoresis in polyacrylamide gel, and five fractions with isoelectric points between pH 4.7 and 5.4 were observed after isoelectric focusing . This microheterogeneity disappeared after denaturation of the enzyme in 0.1% sodium dodecylsulfate or 8M urea . The isoelectric point of the denatured enzyme corresponded to pH 4.3 . All enzyme fractions were identical with regard to their immunological and kinetic properties; they had the same molecular weights . The origin of the different "conformers" of neuraminidase is discussed . The existence of genuine isoenzymes could largely be excluded . The yield of neuraminidase was 65%, which corresponded to about 10 mg of pure enzyme from 100 l of culture medium . The enzyme was free of protease and various other glycosidase activities . The neuraminidase preparation appeared not to be contaminated by other proteins as judged by electrophoretic analysis using either the native enzyme or the enzyme denatured by sodium dodecylsulfate or urea; ultracentrifugation; chromatography on Sephadex G-200; and immunological methods . The molecular weights of the native or denatured enzyme were found to be in the range between 60 000 and 69 000 (on an average 63 750) using four independent methods . The existence of subunits of neuraminidase was excluded . The neuraminidase exhibited a spec . act . of 580 or 615 U/mg protein with glycopeptides from edible birds' nests or sialyllactose, respectively, as substrates . Additional kinetic properties and the UV-absorption spectrum of the enzyme are described.

Appl Microbiol, 1975 Jun, 29(6), 861 - 3
Radiation resistance of spores of some Clostridium perfringens strains; Clifford WJ et al.; Clostridium perfringens spores (eight strains) were irradiated in a model system with 60Co gamma rays at -30 C . The quantal response data obtained were analyzed with extreme value statistics . It was found (at the 95% confidence level) that all eight strains followed the same rate of death and that this rate was probably (at the 95% level) not exponential . The statistics did not exclude, however, a normal, lognormal, Weibull, or related rate of spore kill . A more definitive study would be required to distinguish between the latter distributions.

J Gen Microbiol, 1975 Jun, 88(2), 229 - 44
Taxonomy of the Clostridia: ribosomal ribonucleic acid homologies among the species; Johnson JL et al.; rRNA homologies have been determined on reference strains representing 56 species of Clostridium . Competition experiments using tritium-labelled 23S rRNA were employed . The majority of the species had DNA with 27 to 28% guanine plus cytosine (%GC) . These fell into rRNA homology groups I and II, which were well defined, and a third group which consisted of species which did not belong in groups I and II . Species whose DNA was 41 to 45% GC comprised a fourth group . Thirty species were placed into rRNA homology group I on the basis of having 50% or greater homology with Clostridium butyricum, C . perfringens, C . carnis, C . sporogenes, C . novyi or C . pasteurianum . Ten subgroups were delineated in homology group I . Species in each subgroup either had high homology with a particular reference species or a similar pattern of homologies to all of the reference organisms . The eleven species in rRNA homology group II had 69% or greater homology to C . lituseburense . Species in groups I and II had intergroup homologies of 20 to 40% . The six species in group II had very low homologies with groups I and II . Negligible homology also resulted when five of the species were tested against the sixth, C . ramosum . The five species having DNA with 41 to 45% GC were C . innocuum, C . sphenoides, C . indolis, C . barkeri and C . orotic um . Little rRNA homology was apparent between C . innocuum and the other high % GC species or with several Bacillus species having similar %GC DNA . Correlations between homology results and phenotypic characteristics are discussed.

Biochemistry, 1975 May 20, 14(10), 2146 - 51
Optical spectra and electronic structure of flavine mononucleotide in flavodoxin crystals; Eaton WA et al.; The polarized single-crystal absorption spectra of the oxidized and semiquinone forms of flavodoxin from Clostridium MP have been measured with a double beam recording microspectrophotometer . The spectra establish that the radical species in the crystal is the neutral (blue) falvine semiquinone . Combination of the spectra reported here with polarization data from previous fluorescence and stretched-film studies provides transition moment directions for the first two phi-phi transitions of the oxidized form . Predictions of molecular orbital theory are in good agreement with these experimental directions . The crystal spectra of the semiquinone indicate that the two lowest frequency transitions have the same detailed orbital origin as the corresponding transitions of the oxidized form; in the semiquinone these transitions appear at lower frequency, are closer together, and, as predicted from detailed considerations of transition probabilities, exhibit approximately half the absorption intensity . Our hypothesis of a common orbital origin suggests that semiquinone formation takes place by the addition of an electron to the lowest empty phi orbital of the oxidized form without any gross electronic rearrangement.

Ann Microbiol (Paris), 1975 May-Jun, 126A(4), 421 - 34
{The spore germination of "Clostridium tyrobutyricum" III.--An hypothesis on the mechanism of initiation (author's transl)}; Bergere JL et al.; Spores of C . tyrobutyricum do not contain 3-phosphoglyceric acid (PGA) but a polysaccharide which could replace PGA as an energy source during germination . The absence of PGA, which is an inhibitor of phosphotransacetylase, confirms the role of the acetyl-CoA synthesizing system in the germination initiated by acetate . Spore extracts of C . tyrobutyricum, as extracts of vegetative cells, were found to contain a ferredoxin and exhibited a NADH-ferredoxin oxydase activity which required the presence of an acetyl-CoA regenerating system, suggesting that this enzyme is also involved in germination . From this results, an hypothesis on the role of initiators (acetate and NH4+) in the mechanism of initiation of spore germination in C . tyrobutyricum is proposed . Acetate would have an effect on the utilisation of the endogenous polysaccharide and on glucose catabolism, and therefore, would be an effector for the production of the energy required particularly to transport cations into the spore.

Zentralbl Bakteriol {Orig A}, 1975 May, 231(4), 485 - 90
Feathery outbrusts of the colonies of Clostridium sporogenes containing chain forming mutants after ultraviolet irradiation; Shaikh MR et al.; Cl . sporogenes NCTC 532 was irradiated with UV light, 20 mins . irradiated colonies after initial 48 hours incubation at 37 degrees C and 10-20 days ageing at room temperature (22 to 25 degrees C), gave rise to feathery outbrusts . The material from these outbursts grew in long chains in fluid media . A pure chain - forming culture could not be obtained as the single cells or pairs always "contaminated" the chain - forming culture.

J Gen Microbiol, 1975 May, 88(1), 27 - 35
The preparation of immunogenic cell walls from a highly protective strain of Clostridium chauvoei; Chandler HM et al.; Conventional methods for the preparation of cell walls of a highly protective strain of Clostridium chauvoei destroy the protective antigen . Bacteria were therefore lysed by the enzyme pronase instead of by the mechanical disintegration methods commonly employed . Final purification and separation of cell walls and membranes was achieved by equilibrium density-gradient centrifugation with sodium iodide in a zonal rotor . The resultant cell walls had a two-layered structure when seen in ultra-thin section and were highly immunogenic when used to immunize mice against challenge with C . chauvoei . Rabbit antisera raised against the cell walls provided passive protection against challenge in mice and the level of protection was not diminished by the absorption of all agglutinins from the sera . These results confirm previous observations that the protective antigen is a heatlabile cell wall antigen which stimulates the production of non-agglutinating protective antibody.

J Med Microbiol, 1975 May, 8(2), 299 - 309
Reinvestigation of the taxonomy of Clostridium bifermentans and Clostridium sordellii; Nakamura S et al.; The taxonomic relationships between Clostridium bifermentans and C . sordellii were reinvestigated by numerical taxonomy, studies of DNA-DNA homology and DNA duplex thermal stability, and by analysis of cell-wall sugar components . Although the results indicate that both species may be grouped into one geno-species, C . sordellii strains could be differentiated from C . bifermentans strains on the basis of a few phenetic criteria that include the inability to ferment mannose and sorbitol, the absence of mannose in the cell wall, the production of urease, the absence of arginine deaminase activity, and susceptibility to inhibition of growth by mannose.

J Med Microbiol, 1975 May, 8(2), 289 - 97
Susceptibility to mitomycin C and lecithinase activities of Clostridium oedematiens (C . novyi) type B and D; Nakamura S et al.; In tests with broth-culture products of Clostridium oedematiens, none of 15 type-B strains showed beta-toxin lecithinase activities exceeding 20 egg-units per ml, whereas 12 of 13 type-D strains consistently produced much greater amounts of the lecithinase . The types also differed in their susceptibility to lysis by mitomycin C (MC) . Of 13 type-D strains tested, 12 were sensitive to MC at a concentration of 1 or 2 mug per ml, whilst 14 of 15 type-B strains were insensitive . Phage-like particles were observed in the MC-lysates of some type-D strains . No type-specific differences in the production of indole or the fermentation of maltose were demonstrated.

Zentralbl Bakteriol {Orig A}, 1975 May, 231(4), 451 - 65
{Swarming phenomenon of an aeromonas spec (author's transl)}; Muller HE et al.; A genuine swarming phenomenon, such as has previously been known to occur in Proteus, Bacillus and Clostridium species only, was observed in an Aeromonas species . Fig . 1 shows the terraced swarming zones of the Aeromonas species on nutrient agar . The swarming rate, expressed as the growth of the swarming zone per time unit, was measured to be 70-120 mum/min on blood agar at 30 degrees C . The swarming could be inhibited by incubation at 37 degrees C (Table 2), by low saline concentrations (Table 3) as well as by addition of 4-nitro-phenylglycerol to the medium (Table 4) . A DIENES-phenomenon between the swarming zones of Proteus strains and that of the Aeromonas species could not be observed (Fig . 2) . The manner of swarming as seen in phase contrast microscopy was the same kind as that of Proteus . Furthermore, it could be shown by means of light- and electronmicroscopical investigations that the swarming phenomenon is connected with changes in the cell morphology and the form of flagellation (Figs . 4 and 5) . Whereas in broth cultures (Fig . 3) as well as in the centre of colonies on solid media (Fig . 5a) the cells appeared as cocoid rods with polar flagellation, they developed elongated forms at the edge of the swarming zone, which - either in addition to or devoid of the polar flagella - were peritrichously populated with thin, flagella-like filaments (Figs . tb, 6, 7 and 8) . The discussion deals with the various forms of bacterial surface translocation and investigates into the role of peritrichous flagella or fimbriae in the swarming phenomenon.

J Infect Dis, 1975 May, 131(5), 559 - 66
Production and characterization of exotoxin(s) of Shigella dysenteriae type 1; McIver J et al.; A semicontinuous fermenter system was developed in which broth culture filtrates of Shigella dysenteriae type 1 yielded substantial amounts of exotoxin . Biologic activity of the exotoxin was characterized by means of three assays: the rabbit ileal loop for fluid evocation (enterotoxicity), mouse lethality after parenteral injection (neurotoxicity), and HeLa cell toxicity in vitro (cytotoxicity) . Although the culture filtrate was highly active, disc electrophoresis revealed that the toxin is a minor component of the mixture of proteins in the crude preparation, and that the minor representation contrasts with the relative prominence of exotoxins in cultures of other bacteria, such as Vibrio cholerae, Corynebacterium diphtheriae, and Clostridium tetani . Filtrate toxin, when purified by isoelectric focusing in polyacrylamide gel, was found to contain two toxic moieties . One was resolvable as a single band with an isoelectric joint (pI) of 7.25, a molecular weight of 72,000, and all three types of biological activity . The second moiety was isoelectric at pH 6.00, contained two subcomponents, and further contrasted with the pI 7.25 toxin by being more cytotoxic while being devoid of enteroneurotoxic activity.

Appl Microbiol, 1975 May, 29(5), 598 - 603
Purification and properties of Clostridium botulinum type F toxin; Yang KH et al.; Clostridium botulinum type F toxin of proteolytic Langeland strain was purified . Toxin in whole cultures was precipitated with (NH4)2SO4 . Extract of the precipitate was successively chromatographed on diethylaminoethyl-cellulose at pH 6,0, O-(carboxymethyl) cellulose at pH 4.9, and finally diethylaminoethyl-cellulose at pH 8.1 . The procedure recovered 14 percent of the toxin assayed in the starting culture . The toxin was homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, double gel diffusion serology, and isoelectric focusing . Purified toxin had a molecular weight of 150,000 by gel filtration and 155,000 by sodium dodecyl sulfate-polyacrylamide gel electrophoresis . Specific toxicity was 9.6 X 10-6 mean lethal doses per absorbancy (278 nm) unit . Sub-units of 105,000 and 56,000 molecular weight are found when purified toxin is treated with a disulfide reducing agent and electrophoresed on sodium dodecyl sulfate-polyacrylamide gels . Reciprocal cross neutralizations were demonstrated when purified type F and E toxins were reacted with antitoxins which were obtained with immunizing toxoids prepared with purified toxins.

J Infect Dis, 1975 May, 131 Suppl, S81 - 5
Isolation of Clostridium in human infections: evaluation of 114 cases; Gorbach SL et al.; One hundred fifty-two strains of Clostridium were isolated from 144 patients over a 14-month-peroid . These included 23 recognized species and 23 strains that were unclassified . Soft tissues or abscesses yielded 84 strains of Clostridium . Intraabdominal sites predominated, but clostridia were recovered from empyema, carcinoma, frostbite with gas gangrene, muscle abscess, aortic graft, and brain abscess . Blood cultures yielded 65 strains of Clostridium from 49 patients, representing 0.3% of 16,314 blood cultures (or 2.6% of 2,168 positive cultures) . Clostridium perfringens was most common in blood, accounting for 37 isolates (57%) . Clostridial bacteremia was often unrelated to the clinical setting and was found in alcoholics with aspiration or Streptococcus pneumoniae pneumonia, pulmonary tuberculosis, empyema, meningococcemia, and infantile gastroenteritis . In 20 of the 49 patients (41%), aerobic or other anaerobic bacteria were cultured concurrently from the blood . Thus, clostridial bacteremia should be interpreted with caution since there may be little correlation with the patient's clinical condition.

Infect Immun, 1975 May, 11(5), 932 - 6
Antigenicites of fragments of Clostridium botulinum type B derivative toxin; Kozaki S et al.; Two fragments with molecular weights of 110,000 and 60,000 were separated in a preparatory scale by gel filtration of the reduced Clostridium botulinum type B trypsinized derivative toxin on Sephadex G-200 with 0.05 M tris(hydroxymethyl)aminomethane-0.38 M glycine buffer, pH 8.3, containing 5 mM ethylenediaminetetraacetate, 1 mM dithiothreitol, and 2 M urea as eluant . They were both antigenic, forming crossing precipitin lines against type B antitoxin in agar gel double diffusion tests.

Appl Microbiol, 1975 May, 29(5), 590 - 4
Common mesophilic anaerobes, including Clostridium botulinum and Clostridium tetani, in 21 soil specimens; Smith LD; A relatively rich medium was markedly superior to a dilute medium for the isolation of anaerobic bacteria from soil . The obligate anaerobes isolated from 21 soil samples were all clostridia and the counts ranged from 2.7 X 10-2 to 3.3 X 10-6 per g . The organisms most frequently isolated were Clostridium subterminate, C . sordelii, C . sporogenes, C . indolis, C . bifermentans, C . mangenoti, and C . perfringens . Seventeen other species were also recognized but almost one-third of the isolates could not be identified with any known species of Clostridium . C . botulinum type A was demonstrated in six soil samples, and type B in one . These soils were neutral to alkaline in reaction (average pH 7.9) and low in organic matter content (1.4%) . The association of C . botulinum types A and B with neutral to alkaline soils was statistically significant (P = 0.001) as was their association with soils low in organic matter (P = 0.005) . C . botulinum types E and F were found in one soil sample, pH 4.5, with organic matter 13.7% . C . tetani was isolated from two soil samples, both of intermediate pH value and higher than average organic matter content.

J Med Microbiol, 1975 May, 8(2), 235 - 49
Preparation of a glycoprotein fraction from pooled human plasma and its evaluation as a substrate for the assay of Clostridium welchii (C . perfringens) neuraminidase; Fraser AG et al.; A glycoprotein fraction (fraction VII) suitable for use as a substrate in assays of microbial neuraminidase was prepared from pooled human plasma . It is pasteurised during preparation to eliminate the risk of transmission of serum hepatitis . This results in polymerisation of some of the gamma1-acid glycoprotein, but fraction VII is shown to be an excellent substrate for the neuraminidase of Clostridium welchii (C . perfringens) . A sensitive assay procedure is described . A number of factors may interfere with the measurement of NANA released by the action of microbial neuraminidase and procedures are described for evaluation of this problem . Fraction VII is easy to prepare, cheap and available in standard form in large amounts (inquiries should be addressed to J . K . S.); it is recommended for routine use as a convenient substrate for neuraminidase assays.

Biochem J, 1975 May, 148(2), 279 - 94
Inhibition of glucose 6-phosphatase by pure and impure C-type phospholipases . Reactivation by phospholipid dispersions and protection by serum albumin; Cater BR et al.; 1 . Pure or impure C-type phospholipases hydrolysed rat liver microsomal phosphatides in situ at 5 degrees or 37 degrees C . At 5 degrees C mean hydrolysis of total phospholipids was 90% by Bacillus cereus and 75% by Clostridium perfringens (Clostridium welchii) C-type phospholipases . 2 . Four degrees of inhibition of glucose 6-phosphatase (D-glucose 6-phosphate phosphohydrolase; EC 3.1.3.9) resulted . (a) At 37 degrees C inhibition was virtually complete and apparently irreversible . (b) At 5 degrees C phospholipase C inhibited 50-87% of the activity expressed by intact control microsomal fractions . (c) Bovine serum albumin present during delipidation alleviated most of this inhibition: at 5 degrees C phospholipase C plus bovine serum albumin inhibited by 0-35% (mean 18%):simultaneous stimulation by the destruction of its latency seems to offset glucose 6-phosphatase inhibition, sometimes completely . (d) If latency was first destroyed, phospholipase C plus bovine serum albumin inhibited 30-50% of total glucose 6-phosphatase activity at 5 degrees C . Only this inhibition is likely largely to reflect the lower availability of phospholipids, essential for maximal enzyme activity, as it is virtually completely reversed by added phospholipid dispersions . Co-dispersions of phosphatidylserine plus phosphatidylcholine (1:1, w/w) were especially effective but Triton X-100 was unable effectively to restore activity . 3 . Considerable glucose 6-phosphatase activity survived 240min of treatment with phospholipase C at 5 degrees C, but in the absence of substrate or at physiological glucose 6-phosphate concentrations the delipidated enzyme was completely inactivated within 10min at 37 degrees C . However, 80mM-glucose 6-phosphate stabilized it and phospholipid dispersions substantially restored thermal stability . 4 . It is concluded that glucose 6-phosphatase is at least partly phospholipid-dependent, and complete dependence is not excluded . For reasons discussed it is impossible yet to be certain which phospholipid class(es) the enzyme requires for activity.

J Med Microbiol, 1975 May, 8(2), 251 - 63
The production of neuraminidase by food poisoning strains of Clostridium welchii (C . perfringens); Fraser AG et al.; The production of neuraminidase by a classical strain of Clostridium welchii (C . perfringens) type A was studied . Good yields were produced in 5% Proteose Peptone-water medium (PPW5); the enzyme was essentially extracellular but some further neuraminidase could be released by ultrasonic disintegration of the cells . This also released N-acyl neuraminic acid-aldolase (NAN-aldolase) and the degree to which this interferes with the assay for neuraminidase was evaluated . Forty-one British reference food-poisoning strains of C . welchii type A were examined for extracellular neuraminidase production in PPW5 . Twelve of 17 strains that produce so-called heat-sensitive spores were neuraminidase positive whereas 20 of 24 strains that are non-haemolytic and produce very heat-resistant sporeswere neuraminidase negative . Variation was found in the ability to produce neuraminidase among strains of a single Hobbs' serotype; four Hobbs' type-13 strains produced neuraminidase but a fifth did not . Disruption of the cells of a Hobbs' type-2 strain that did not produce any extracellular neuraminidase released NAN-aldolase but there was no evidence of cell-associated neuraminidase . British food-poisoning strains of C . welchii type A thus include some that are clearly neuraminidase positive and some that still cannot be shown to produce neuraminidase . There is no correlation between lack of neuraminidase production and the ability to cause food poisoning, although the majority of non-haemolytic heat-resistant strains do not produce neuraminidase . It remains possible that neuraminidase may play a part in C . welchii gas gangrene; it is suggested that the ability to define neuraminidase-negative strains may now be of value in investigating this possibility.

Infect Immun, 1975 May, 11(5), 1061 - 8
Characterization of enterotoxin purified from Clostridium perfringens type C; Skjelkvale R et al.; Enterotoxin produced by a sporulating culture of Clostridium perfringens type C, which had been isolated from a case of severe necrotic enteritis, was purified . The molecular weight was estimated to be 36,000 by gel chromatography on Sephadex G-100 and 33,400 by ultracentrifugation . The sedimentation coefficient S20,W was 2.92 . The toxin protein exhibited unusual behavior on sodium dodecyl sulfate gels, and toxin aggregates having molecular weights of 68,000, 85,000, 105,000, and 140,000 were obtained . The purified enterotoxin often separated, apparently due to slight charge differences, into two protein bands on 7% polyacrylamide gels . Electrofocusing of enterotoxin on polyacrylamide gels gave an approximate isoelectric point of 4.3, with the enterotoxin being fractionated into four distinct protein bands . The specific toxicity of the enterotoxin was about 1,900 mouse mean lethal doses per mg of calculated nitrogen . The data obtained indicate that the enterotoxin from C . perfringens type C is identical in most respects to that obtained from type A strains . Whether or not this toxin plays a role in the necrotic enteritis caused by type C strains is unknown at present.

Biochim Biophys Acta, 1975 Apr 8, 382(4), 479 - 93
Interaction of Clostridium perfringens theta-haemolysin, a contaminant of commercial phospholipase C, with erythrocyte ghost membranes and lipid dispersions . A morphological study; Smyth CJ et al.; Commercially available preparations of phospholipase C from Clostridium perfringens are commonly contaminated with theta haemolysin, one of a group of bacterial haemolysins called oxygen labile (O-labile) haemolysins . Treatment of erythrocyte ghosts and a mixed lipid dispersion containing cholesterol with commercially available phospholipase C in the absence of Ca-2+ and the presence of phosphate buffer and/or EDTA resulted in the formation and release of ring or arc-shaped structures . Highly purified phospholipase C, free of theta-haemolysin, produced no changes in the morphology of erythrocyte ghosts or lipid dispersions in the presence of phosphate or EDTA, but caused the formation of typical diglyceride droplets in the presence of Ca-2+ in the absence of these inhibitors . Ring structures, identical to those caused by commercial phospholipase C, were formed on addition of highly purified theta-haemolysin to erythrocyte ghost membranes, lipid dispersions containing cholesterol and cholesterol dispersions, but not on treatment of membranes from Micrococcus lysodeikticus . Heat-inactivated O-haemolysin (60 degrees C for 10 min) produced no such effects . The dimensions of rings and arcs displayed heterogeneity . The outside diameters in various preparations varied from approx . 27-58 nm with border thickness of 4.1-7.8 nm.

Biochemistry, 1975 Apr 8, 14(7), 1390 - 6
Studies on interaction between histone V (f2c) and deoxyribonucleic acids; Hwan JC et al.; Histone V (2fc) from chick erythroctes was used in the study of its interaction with DNA from various sources . Complexes between this histone and DNA were formed using the procedure of continuous NaCl gradient dialysis in urea . Two physical methods, namely thermal denaturation and circular dichroism (CD), were used as analytical tools . Thermal denaturation of nucleohistone V with chick or calf thymus DNA shows three melting bands: band I at 45-50 degrees corresponds to free base pairs; band II at 75-79 degrees, and band III at 90-93 degrees correspond to histone-bound base pairs . In histone-bound regions, there are 1.5 amino acid residues/nucleotide in nucleohistone V . In contrast, a value between 2.9 and 3.3 was determined for nucleohistone I (fl) (H . J . Li (1973), Biopolymers 12, 287) . Similar melting properties have been observed for histone V complexed with bacterial DNA from Micrococcus luteus . Histone V binding to DNA induces a slight transition from a B-type CD spectrum to a C-type spectrum . Trypsin treatment of nucleohistone V reduces melting band III much more effectively than band II . Such a treatment also restores DNA to B conformation in the free state . Reduction of the melting bands of nucleohistone V by polylysine binding follows the order of I greater than II greater than III, accompanied by the increase of a new band at 100 degrees . When two bacterial DNAs of varied A + T (adenine + thymine) content simultaneously compete for the binding of histone V, the more (A " T)-rich DNA is selectively favored . Under experimental conditions described here, Clostridium perfringens DNA with 69% A + T is bound by histone V in preference to chicken DNA with 56% A + T although the latter has natural sequences for histone V binding.

Biochim Biophys Acta, 1975 Apr 8, 382(4), 565 - 75
Hydrolysis of erythrocyte membrane phospholipids by a preparation of phospholipase C from Clostridium Welchii . Deactivation of (Ca-2+, Mg-2+)-ATPase and its reactivation by added lipids; Coleman R et al.; 1 . Haemoglobin-free erythrocyte ghosts were prepared in 40 imosM bicarbonate buffer, pH 7.4, containing 1 mM EDTA (40 imosM/l mM EDTA) . The ghost preparation was highly permeable on preparation but partially resealed on incubation in media containing Ca-2+ . 2 . A partially purified preparation of phospholipase C from Clostridum welchii caused an increase in observed Mg-2+-ATPase activity, reflecting a change in the permeability of the ghost to substrate . The phospholipase did not decrease Mg-2+-ATPase even at the highest levels tested . Mg-2+-ATPase activity could therefore be used as a permeability indicatior in these experiments . 3 . Both (Ca-2+, Mg-2+)-ATPase activities of the ghosts were progressively lost as a result of the phospholipid hydrolysis induced by phospholipase C . 4 . When a haemolysin in the commercial preparation was destroyed by heat-treatment, deactivation of the (Ca-2+, Mg-2+)-ATPase and (Na+, K+, Mg-2+)-ATPases were still observed but permeability changes were greatly reduced . 5 . The products of phospholipase action were not inhibitory to the Ca-2+, Mg-2+)-ATPase . 6 . Lysolecithin brought about a reactivation of the (Ca-2+, Mg-2+)-ATPase which was superimposed upon permeability changes in the preparation . 7 . Reactivation of the (Ca-2+, Mg-2+)-ATPase was brought about by a nonlytic, mixed lipid preparation without significant effect upon permeability . 8 . Human erythrocyte (Ca-2+, Mg-2+)-ATPase therefore appears to be an enzyme which responds to perturbation of the lipid environment in the membrane and is a "lipid-dependant" enzyme.

Ann Microbiol (Paris), 1975 Apr, 126(3), 295 - 314
{An intracellular polysaccharide involved in sporulation of "Clostridium butyricum" I . Cytology, production and preliminary enzymic analysis (author's transl)}; Bergere JL et al.; Free glucose concentration and polysaccharide production in Clostridium butyricum cells have been studied with an enzymatic method . Results indicated a substantial decrease in intracellular glucose content simultaneously with a production of polysaccharide prior to the end of exponential growth . Then the polysaccharide accumulated rapidly to reach a maximum just before the first refractile spores appeared, and it decreased by 50% during the last stages (V and VI) of sproulation . Electron micrographs of ultrathin sections have demonstrated that most of the polysaccharide is located inside the mother cell cytoplasm as large granules when the remaining is dispersed within the spore cytoplasm beginning during stage III of the sporulation . Overall results showed that production and use of C . butyricum polysaccharide were closely related to sporulation . The isolated polysaccharide exhibited poor water solubility, iodine spectrum with a lambda max at 545 nm and 72% beta-amylolysis . Total hydrolysis occurred with amyloglucosidase indicating an alpha-glucan containing alpha(1 leads to 4) and alpha(1 leads to 6) glucose linkages . The debranching from its beta-dextrin limit by pullulanase revealed the presence of a glycogen like-type structure with some external chains which are longer than those of a normal glycogen . This glycogen arrangement appeared to be of clusters linked by linear chains at least as long as the longest external chains.

Appl Microbiol, 1975 Apr, 29(4), 444 - 7
Molecular construction of Clostridium botulinum type F progenitor toxin; Ohishi I et al.; Molecular dissociation of purified type F progenitor toxin with an S20,W of 10.3 and a molecular weight of 235,000 into two components, toxic and atoxic, was demonstrated by ultracentrifugation, gel filtration, and diethylaminoethyl-Sephadex chromatography at pH 7.5 . The ultracentrifugal analysis indicated that type F progenitor toxin dissociates into components of the same molecular size of 5.9S . The toxic component contained a toxicity of 2.5 times 10-8 50% lethal doses per mg of N . Much higher stability of progenitor toxin than that of derivative toxin, particularly at pH below 5, suggests that only progenitor toxin can act as an oral toxin.

J Gen Microbiol, 1975 Apr, 87(2), 219 - 38
The identification and purification of multiple forms of theta-haemolysin (theta-toxin) of Clostridium perfringens type A; Smyth CJ; The theta-haemolysin of Clostridium perfringens was purified from culture supernatant fluids of type A strains by fractional ammonium sulphate precipitation and isoelectric focusing in narrow pH 5 to 8 gradients . Four components detected on electrofocusing were designated theta-1(pI6-8to6-9),theta-2(pI6-5to6-6),theta-3(pI6-1to6-3) and theta-4(pI5-7to5-9) . Specific activities ranged from 0-4 x 10-6 to 1-2 x 10-6 haemolytic units/mg protein and 2950 to 3600 LD-50/mg protein . Each haemolytic component was activated by cysteine hydrochloride, and inactivated by cholesterol, by addition of sheep erythrocyte ghosts and by heating at 60 degrees C for 10 min; mouse erythrocytes were more resistant than sheep erythrocytes to haemolysis . A reaction of identity was obtained between components in gel diffusion . Sodium dodecyl sulphate polyacrylamide discgel electrophoresis gave molecular weights in the range 59,000 to 62,000 for each component . A similar value was obtained for theta-1 on density gradient ultracentrifugation . Although the multiple forms were free of 11 factors present in culture supernatants, crossed immunoelectrophoresis and disc gel electrophoresis revealed minor contaminants . These studies reveal that theta-haemolysin has physical properties in common with other oxygen-labile haemolysins.

Am J Vet Res, 1975 Apr, 36(4 Pt 2), 583 - 5
Ulcerative enteritis--clostridial antigens; Berkhoff GA; The causative agent of ulcerative enteritis (UE) is a clostridium and it is believed that it belongs to a new, hitherto undescribed species . It has been named, tentatively, Clostridium colinum . Transmission experiments carried out with artificial cultures of this clostridium have clarified some aspects of the pathogenesis of UE and have brought into focus other aspects of its pathogenesis, epizootiology, and immunology that need to be investigated . Support for research is needed: 1) To investigate the distribution in nature of C colinum . For this purpose the development of more suitable selective and differential culture mediums should be encouraged . 2) To further study the pathogenesis of UE through application of the fluorescent labeled antibody procedure . 3) To search for toxins produced by this clostridium, possibly through use of ligated intestinal loops in chickens . 4) To develop a quick and simple means for the differential diagnosis of UE . Use of a fluorescent antibody test has been suggested as an area of research to fill this need.

Infect Immun, 1975 Apr, 11(4), 640 - 8
Determination of toxin-induced leakage of different-size nucleotides through the plasma membrane of human diploid fibroblasts; Thelestam M et al.; Human diploid lung fibroblasts were treated with cytolytic bacterial toxins and the nature of the membrane damage was investigated . {3H} uridine was used for differential labeling of cytoplasmic components of small or large molecular size . Two principal size categories were achieved by labeling the fibroblasts in either early growth phase or stationary phase, a high-molecular weight ribonucleic acid label and a low-molecular-weight nucleotide label . The size of the labeled molecules was determined by perchloric acid precipitation and gel chromatography . Leakage of labeled molecules of different size indicated the size of the "functional pores" in the plasma membrane caused by the test substance . The nonionic detergent Triton X-100 produced large functional pores in the fibroblast membrane as evidenced by rapid leakage of both large and small labeled molecules . Theta-toxin from Clostridium perfringens and the polyene antibiotic filipin both gave rise to considerably small functional pores in the plasma membrane . Although small molecules easily passed the treated membrane, large molecules could not escape from the cells even after prolonged treatment with these substances or by increasing their concentration . By the contrast, the leakage profiles obtained with melittin from bee venom or with delta-toxin from Staphylococcus aureus in each case suggested the formation initially of pores of intermediate size that increased upon prolonged incubation or when higher concentrations were used.

Biochim Biophys Acta, 1975 Mar 28, 386(1), 80 - 6
Application of affinity chromatography to the purification of collagenase for the isolation of insoluble elastin; Serafini-Fracassini A et al.; ClostrIdium histolyticum collagenase (clostridiopeptidase A, EC 3.4.4.19) was purified by batchwise separation with DEAE-cellulose followed by affinity chromatography on a column of alkali-treated elastin . The N-terminal amino acid profile of elastin isolated from bovine ligamentum nuchae using this enzyme preparation was compared with that of a duplicate sample purified with a mixture of collagenase I and II (Yoshida, E, and Noda, H . (1965) Biochim . Biopsys . Acta 105, 562-574) . An approx . three-fold decrease in the molar concentration of N-terminal residues and a considerable reduction in their number was obtained by using the former enzyme preparation.

J Biol Chem, 1975 Mar 25, 250(6), 2062 - 72
The use of 13C nuclear magnetic resonance of aromatic amino acid residues to determine the midpoint oxidation-reduction potential of each iron-sulfur cluster of Clostridium acidi-urici and Clostridium pasteurianum ferredoxins; Packer EL et al.; 13C NMR of the aromatic residues of Clostridium acidi-urici {Phe2}ferredoxin (a chemically modified ferredoxin in which a phenylalanyl residue replaces a tyrosyl residue) and Clostridium pasteurianum ferredoxin permits one to distinguish and probe each iron-sulfur (Fe7S7) cluster neighboring the aromatic residues within each protein . This is because the ring carbon resonance shifts of the phenylalanyl and tyrosyl residues can be distinguished . The 13C NMR results suggest that the midpoint oxidation-reduction potentials of the two Fe4S4 clusters in C . pasteurianum and C . acidi-urici ferredoxin differ by 10 plus or minus 5 mv and smaller than mv, respectively . 13C NMR of an equilibrium mixture of methyl viologen-reduced C . acidi-urici and C . pasteurianum ferredoxin shows that the protein midpoint oxidation-reduction potential of C . acidi-urici ferredoxin is 47 plus or minus 10 mv lower than that of C . pasteurianum ferredoxin . We attribute the differences in cluster and protein midpoint oxidation-reduction potentials to differences in protein structure.

Vopr Med Khim, 1975 Mar-Apr, 21(2), 143 - 7
{Ornithine transcarbamylase in Clostridium perfingens}; Dolgikh MS et al.; Five strains of A type Cl . perfringens of different titres of toxicity (BP6K, SR-12, 2836, 2910, 1) possessed a distinct activity of ornithine transcarbamylase . In cells the maximal activity of the enzyme was observed within 3-5 hrs of growth of the culture . The enzyme was partially purified and some of its physico-chemical properties were studied . Ornithine transcarbamylase was termostable . Monoiodine acetate, p-chloromercurybenzoate, semicarbazide, Hg-2+, Cu-2+ and Fe-3+ inhibited the enzymatic activity, but Mg-2+ and Ca-2+ activated the enzyme . Ornithine transcarbamylase was shown to be active in wide region of pH: from pH 6.0 to pH 9.0 and higher . Two peaks of the enzyme activity were observed: the first (a more distinct one) at pH 8.6-8.9 and the second, smaller,--at pH 6.5-6.7.

Can J Microbiol, 1975 Mar, 21(3), 235 - 44
Stable isotope fractionation by Clostridium pasteurianum . 1 . 34S/32S: inverse isotope effects during SO4-2- and SO3-2- reduction; McCready RG et al.; During growth on minimal salts--sucrose media supplemented with various concentrations (10-4-10-2 M) of sodium sulfate, Clostridium pasteurianum grew at a normal rate and only evolved sulfide in late stages of growth on 10-2 M SO4-2- . The evolved sulfide was slightly enriched in 34S as compared to the medium sulfur . On the other hand, sulfide was evolved during growth on all concentrations of sulfite tested . Large normal and inverse isotopic effects were observed in the evolved sulfide during SO3-2- reductions . In contrast, the intracellular sulfur showed much smaller fractionations . The complexity of the isotopic patterns suggests that a dissimilatory sulfite reductase system may be induced by high concentrations of sulfite.

Onderstepoort J Vet Res, 1975 Mar, 42(1), 25 - 7
Effect of Clostridium perfringens epsilon toxin on the blood brain barrier of mice; Worthington RW et al.; It was shown that Clostridium perfringens epsilon toxin has the effect of allowing the passage of 125I polyvinyl-pyrrolidone and 125I human serum albumin into mouse brain . These substances did not enter the brains of normal control mice . The passage of albumin into the brains of mice poisoned with epsilon toxin was extremely rapid . When large doses of toxin (+/-4 000 MLD) were given death ensued within 2-3 min at which stage 1,5% of the injected albumin had already entered the brain . In cases where smaller doses were given and the time interval between injection and death was longer the figure was increased to 2-2 1/2% of the injected plasma albumin.

Nord Vet Med, 1975 Mar, 27(3), 150 - 2
{Acute bovine mastitis caused by clostridium perfringens type A (Author's transl.)}; Nesbakken T et al.; An acute case of bovine mastitis caused by clostridium perfringens type A is described . The condition appeared before delivery and was characterised by gas formation in the udder and severe general signs . In spite of parenteral treatment with large doses of penicillin and streptomycin combined with frequent stripping of the udder the animal died two days after the onset of symptoms . The strain isolated proved sensitive to penicillin, chloramphenicol and tetracyclin, but resistant to streptomycin and sulphonamides.

J Gen Microbiol, 1975 Mar, 87(1), 120 - 8
The physiological function of nitrate reduction in Clostridium perfringens; Hasan SM et al.; Fermentation-balance studies have been carried out on Clostridium perfringens grown in the presence and absence of nitrate in the medium . Nitrate is able to serve as an electron acceptor for these bacteria, permitting increased growth yields over those obtained in its absence . This increase is due to an increase in the proportion of metabolite molecules which can participate in substrate-level phosphorylation reactions when an inorganic acceptor is available . The nitrate reduction can be regarded as a primitive form of anaerobic respiration in these bacteria, since it is clearly coupled to their energy metabolism and is not assimilative in function . We believe that the existence of this kind of energy metabolism in these bacteria has significant evolutionary implications.

Infect Immun, 1975 Mar, 11(3), 563 - 75
Enterotoxin formation by different toxigenic types of Clostridium perfringens; Skjelkvale R et al.; Sixty-nine strains of Clostridium perfringens of different toxigenic types were investigated for enterotoxin production . Enterotoxin was definitively detected only in strains of types A and C . This is the first report where enterotoxin production has been demonstrated in a toxigenic type other than type A . The exterotoxin-positive type C strains were isolated from cases of enteritis necroticans ("pig bel+) in New Guinea . The major enterotoxin from type C showed a reaction of complete identity with enterotoxin from type A in immunodiffusion using anti-enterotoxin serum prepared against the latter; it induced erythema when injected intradermally into depilated guinea pigs and caused fluid accumulation in the rabbit ileal loop . The results indicate that the major enterotoxin from type C was serologically and biologically similar to enterotoxin from type A . In some C was serologically and biologically similar to enterotoxin from type A . In some type C strains, an enterotoxin was detected that showed a reaction of partial serological identity . Spore coat proteins were extracted from 14-strains by alkaline dithiothreitol, and the extracts were assayed for enterotoxin-like spore protein . Enterotoxin could be extracted from type A and type C spores, and all positive strains showed a reaction of complete identity in immunodiffusion with enterotoxin obtained from cell extracts of type A . Disc immunoelectrophoresis demonstrated that two distinct components that reacted serologically with anti-enterotoxin serum were present in both the cell extract and in extracted spore protein from one type C strain . These distinct components differed in molecular weight.

Infect Immun, 1975 Mar, 11(3), 526 - 9
Analysis of unidirectional fluxes of sodium during diarrhea induced by Clostridium perfringens enterotoxin in the rat terminal ileum; McDonel JL et al.; Net intestinal transport of sodium in vivo, in control and enterotoxin (Clostridium perfringens)-treated rats, was resolved into two unidirectional fluxes, influx from and efflux into the lumen of the terminal ileum . In rats treated with the toxin, sodium influx remained similar to control values even during fluid and electrolyte loss to the lumen . Net loss of sodium was shown to be due to nearly a twofold increase in sodium efflux to the lumen in toxin-treated animals . There was only slight histopathological damage to the mucosa, especially noticeable at the tips of villi.

Appl Microbiol, 1975 Mar, 29(3), 345 - 51
Influence of carbohydrates on growth and sporulation of Clostridium perfringens type A; Labbe RG et al.; Growth and sporulation of Clostridium perfringens type A in Duncan and Strong (DS) sporulation medium was investigated . A biphasic growth response was found to be dependent on starch concentration . Maximal levels of heat-resistant spores were formed at a starch concentration of 0.40% . Addition of glucose, maltose, or maltotriose to a sporulating culture resulted in an immediate turbidity increase, indicating that biphasic growth in DS medium may be due to such starch degradation products . Amylose and, to a lesser extent, amylopectin resulted in biphasic growth when each replaced starch in the sporulation medium . A levels of heat-resistant spores approximately equal to the control was produced with amylopectin but not amylose as the added carbohydrate . Addition of glucose or maltose to a DS medium without starch at stage II or III of sporulation did not alter the level of heat-resistant spores as compared with the level obtained in DS medium with starch . Omission of starch or glucose or maltose resulted in an approximately 100-fold decrease in the number of heat-resistant spores, although the percentage of sporulation (90%) was unaffected . The role of starch and amylopectin in the formation of heat-resistant spores probably involves the amyloytic production of utilizable short-chain glucose polymers that provide an energy source for the completion of sporulation.

Lancet, 1975 Feb 22, 1(7904), 420 - 1
Clostridium defficiel in the urogenital tract of males and females; Hafiz S et al.; A study of the occurrence of Clostridium difficile in the urogenital tract of males and females revealed higher isolation-rates in patients attending the special (venereal-disease) clinic than in patients attending family-planning and urological clinics . The presence of Cl . difficile in patients with venereal diseases is being investigated to see if the organism is simply an opportunist infecting a urethra disturbed by some antecedent disease, or if it is perhaps a primary cuase of disease.

Nouv Presse Med, 1975 Feb 15, 4(7), 483 - 6
{Septicemias of superinfection in resucitation . Their prevention by restriction of antibiotics}; Rapin M et al.; Nosocomial septicemias supervened during the 15 trimesters following the opening of a new built critical care unit were reviewed . During the five first trimesters the incidence of nosocomial septicemias was 5.9 p.cent . It increases to 10.7 p.cent (p smaller than 0.001) during the five following trimesters . Then a drastic restriction in antibiotic administration was performed, including withdrawal of any prophylactic antibiotherapy, preferential use of penicilline G to prevent infection by Clostridium and cocci Gram positive species surinfection, and, in case of bacterial proved infection, use of a narrow spectrum adapted antibiotic for a time as short as possible . During the 5 trimesters following the application of this antibiotic restriction, the incidence of nosocomial septicemias dropped to 5.7 p.cent (p smaller than 0.001) . This phenomenon was related to the lowered incidence of the Gram negative bacilli septicemias from 11.1 p.cent to 4.8 p.cent (p smaller than 0.001) while the incidence of Gram positive cocci septicemias remained unchanged . These results strongly suggest the responsibility of the overuse of antibiotics in the determination of nosocomial septicemias and the efficiency of a drastic antibiotic restriction upon the surinfection of patients hospitalized in a critical care unit.

J Med Microbiol, 1975 Feb, 8(1), 167 - 72
A defined medium for the growth of Clostridium tetani and other anaerobes of clinical interest; Watt B et al.; The growth of six strains of Clostridium tetani was assessed in a chemically supplemented commercially available defined medium . All strains grew reliably even after 12 serial passages, and two strains produced demonstrable toxic activity after passage . Consistent growth of the test strains could also be obtained on a solid version of this medium ("CA109-S" medium), and the strains could be serially passaged on this medium . Preliminary evidence is presented that the medium supports the surface growth of some other test anaerobes . Such a defined solid medium might prove of value in further studies on the surface growth of C . tetani and of other anaerobes of clinical interest.

J Hyg (Lond), 1975 Feb, 74(1), 1 - 6
Clostridium botulinum in Scottish fish farms and farmed trout; Burns GF et al.; Rainbow trout and specimens of pond mud were collected from three fish farms and examined for the presence of Clostridium botulinum . Two of the farms were constructed with concrete channels and one was mud-bottomed . Cl . botulinum was isolated only from the mud-bottomed farm (24% of muds), and the isolates were all non-proteolytic type B . The implications of the presence of Cl . botulinum spores in the mud of fish farms is discussed.

Can J Microbiol, 1975 Feb, 21(2), 221 - 6
Comparison of the effects of three fluorocarbons on certain bacteria; Van Auken OW et al.; Three fluorocarbons were tested to determine their effect on bacterial growth . Freon 11 and 21 in various concentrations had an inhibitory effect on selected test organisms, but Freon 22 had no effect . Both aerobic and anaerobic microorganisms, as well as gram-positive and gram-negative species, were included among the bacteria tested . Freon 11 and 21 caused a similar response with Freon 11 being more inhibitory to some species and Freon 21 more inhibitory to others . Inhibition was dependent on the concentration of the halocarbon and resulted in decreased respiration rates at all concentrations tested . Results reported here indicate that the action of the fluorocarbons tested is bactericidal rather than bacteriostatic, Serratia marcescens and Clostridium botulinum were the species most sensitive to the halocarbons tested.

Am J Vet Res, 1975 Feb, 36(2), 181 - 5
Grain overload in cattle and sheep: changes in microbial populations in the cecum and rumen; Allison MJ et al.; Samples from the rumen and cecum of cattle and sheep were cultured to determine changes in microbial populations resulting from overfeeding with grain . Before the animals were overfed, the predominant organisms from both sides were those that grew anaerobically on a relatively nonselective ruminal fluid medium and would not grow on selective mediums designed to culture lactobacilli, streptococci, coliforms, or Clostridium perfringens . By 24 hours after overfeeding, lactic acid bacteria had increased in numbers so that they were the most numerous organisms in both the rumen and the cecum . The concentrations of coliforms and C perfringens also increased after overfeeding and were generally higher in the cecum than in the rumen.

J Infect Dis, 1975 Feb, 131(2), 182 - 5
Susceptibility of anaerobic bacteria to metronidazole: relative resistance of non-spore-forming gram-positive baccilli; Chow AW et al.; Susceptibility of 358 clinical isolates of obligate anaerobes to metronidazole was determined by an agar-dilution technique . Only 66% of all isolates were inhibited by 6.25 mug/ml, whereas 30% required larger than or equal to 50 mug/ml . Considerable variation in susceptibility was observed among different genera and species of bacteria . Fusobacterium was most senstitive, followed by Clostridium, Bacteroides and Peptococcus, Peptostreptococcus, Veillonella and Acidaminococcus, and non-spore-forming gram-positive bacilli . Bacteroides fragilis was more sensitive than other species of Bacteroides . Similarly, Clostridium perfringens was more susceptible than other species of Clostriduim . While metronidazole appears to be a promising antimicrobial agent for infections caused by Fusobacterium, Clostrididium, and B.fragilis, therapy for infections with other anaerobic bacteria should be guided by in vitro tests of sensitiivity.

Appl Microbiol, 1975 Feb, 29(2), 145 - 51
Collagenolytic activity of some marine bacteria; Merkel JR et al.; Reconstituted, acid-extracted collagen was used to prepare a medium to screen proteolytic marine bacteria for their ability to elaborate collagenolytic enzymes . The medium was resistant to solubilization by trypsin, hyaluronidase, chondroitinase ABC, and various marine proteinases, but was readily hydrolyzed by commercial Clostridium collagenases . Eighty-seven marine isolates collected in the vicinity of Bermuda, Oahu (Hawaii), and Stone Harbor and Cape May, N . J., were screened . Approximately 44 per cent of the isolates were capable of elaborating enzymes that hydrolyzed reconstituted collagen gels . Several cultures produced collagenolytic enzymes only when grown in the presence of collagen or degradation products of collagen, and with very few exceptions the presence of collagen in the medium greatly enhanced collagenolytic enzyme production . The enzymes from a collagenolytic Bermuda marine isolate were studied in more detail to illustrate that the enzymes capable of hydrolyzing reconstituted collagen were separable from nonspecific proteinases by zone electrophoresis and that these enzymes were true collagenases by virtue of their ability to hydrolyze native bovine Achilles'tendon obtained from three different sources.

Can J Microbiol, 1975 Feb, 21(2), 181 - 5
Composition of the capsular polysaccharides of Clostridium perfringens as a basis for their classification by chemotypes; Paine CM et al.; An analytical procedure, using gas-liquid chromatography, was developed for the identification of the per(trimethylsilyl) ethers of the constituent monosaccharides obtained from the capsular polysaccharides of Clostridium perfringens Hobbs 5, Hobbs 9, Hobbs 10, and NCTC 10578 . Qualitative and quantitative differences between the major polysaccharide components enabled the differentiation of the four strains of C . perfringens investigated.

Cancer Res, 1975 Feb, 35(2), 287 - 90
Colon carcinogenesis with azoxymethane and dimethylhydrazine in germ-free rats; Reddy BS et al.; The effect of intestinal microflora on the sensitivity of the colon to the carcinogenic effect of azoxymethane and a large dose of 1,2-dimethylhydrazine was studied using germ-free and conventional female Fischer rats . Injection s . c . of 1,2-dimethylhydrazine-induced tumors of the ear duct, kidney, and small intestine of conventional rats but none in germ-free animals . Only 20% germ-free rats showed 1,2-dimethylhydrazine-induced colonic tumors, whereas 93% of conventional rats developed multiple colonic tumors . Intrarectal instillation of azoxymethane appreciably increased the multiplicity of colonic tumors in germ-free rats and in gnotobiotic rats contaminated with Clostridium perfringens, as compared to conventional controls . None of the germ-free rats showed ear duct tumors . The incidence of kidney tumors was lower in germ-free rats than in other groups . It is concluded than the intestinal microbial populations alter the effect of carcinogens in the large intestine.

Jpn J Microbiol, 1975 Feb, 19(1), 1 - 6
Immunochemical characterization of cell wall protein antigen purified from the cell wall autolysate of Clostridium botulinum type A; Takumi K et al.; The cell wall protein antigen was solubilized from the isolated cell walls of Clostridium botulinum type A by autolysis and purified by diethylaminoethyl-cellulose column chromatography followed by gel filtration on Sephadex G-150 . The two fractions showed a high degree of the serological activity and produced a main fused precipitin line in immunodiffusion tests against the homologous antiserum . The fact that antigenic fractions contained various kinds of amino acids but no detectable amounts of amino sugars or carbohydrates suggests that the antigens were principally composed of proteins . The protein antigen possessed multiple antigenic components in immunoelectrophoresis . As serological activity, the antigen was heat-stable and resistant to tryptic digestion but sensitive to the actions of pronase, nagarse or pepsin . The protein antigen appeared to be responsible for the common antigenicity among the proteolytic strains of C . botulinum.

Can J Microbiol, 1975 Feb, 21(2), 146 - 51
Immunophyletic relationships between actinomycetales and eubacteriales determined by examinations of their cytoplasmic antigens; Kwapinski JB et al.; Cytoplasms obtained from a total of 205 different strains of Actinomycetales, Eubacteriales, Lower Fungi, and Protozoa were examined against selected anti-cytoplasm antisera using the immunodiffusion and counterimmunoelectrophoresis procedures . Immunological relationships between the different genera have been revealed and, in respect to Eubacteriales, the possible immunophyletic determinants have been discerned . Cytoplasms of Enterobacter, Clostridium, Staphylococcus, and Diplococcus were found to retain the significant linkage determinants between large groups of Eubacteriales and Actinomycetales . Evolutionary implications of the findings are discussed.

Biochim Biophys Acta, 1975 Jan 31, 376(1), 63 - 71
The iron electron-nuclear double resonance (ENDOR) of 4-Fe clusters in iron-sulfur proteins from Chromatium and Clostridium pasteurianum; Anderson RE et al.; Iron electron-nuclear double resonance (ENDOR) measurements were made of the 4-Fe clusters in oxidized Chromatium high-potential iron-sulfur protein, dithionite-reduced high-potential iron-sulfur protein in 80% dimethylsulphoxide, fully reduced Clostridium pasteurianum ferredoxin in aqueous solution and in 80% dimethylsulfoxide . The hyperfine couplings determined show that: i) the electron distribution in each case is nearly symmetric; ii) there are two types of iron in oxidized high potential iron-sulfur protein; iii) only one type of iron is observed in each fully reduced 4-Fe cluster; iv) the data also suggest a greater electron delocalization onto the ligands as compared to the 2-Fe ferredoxins.

J Biol Chem, 1975 Jan 10, 250(1), 261 - 70
Electron paramagnetic resonance and water proton relaxation rate studies of formyltetrahydrofolate synthetase-manganous ion complexes . Evidence for involvement of substrates in the promotion of a catalytically competent active site; Buttlaire DH et al.; Conformational properties of the active site of formyltetrahydrofolate synthetase from Clostridium cylindrosorum have been examined by EPR spectroscopy and by solvent proton relaxation rate (PPR) studies of manganous complexes with the enzyme . Ternary enzyme-Mn-nucleotide complexes give EPR spectra which are very similar to those for the binary Mn-nucleotide complexes . However, upon addition of tetrahydrofolate to form the quaternary complexes, enzyme-MnADP-tetrahydrofolate and enzyme MnATP-tetrahydrofolate the EPR line shapes are changed substantially . Spectra for the quaternary complexes exhibit narrow line widths, and the splitting patterns are characteristic of a slightly asymmetric electronic environment for the bound Mn(II) . Addition of formate to the ADP quatenary complex induces a further significant narrowing of the EPR line widths, although in the absence of tetrahydrofolate, formate does not influence the EPR spectrum for the enzyme-MnADP species . Both Pi and nitrate cause changes in the EPR patterns for the higher complexes of the enzyme which involve both ADP and tetrahydololate . However, the Pi effect is not influenced by the presence of formate whereas the characteristic effect of nitrate is potentiated only when formate is present . EPR sectra for the thernary complex with the beta, gamma-methylene analog of ATP App(CH2)p differ significantly from spectra for the binary App(CH)p complex is not influenced by further additions of tetrahydrofolate and of tetrahydorfolate and formate . The failure of spectra for the App(CH)p complex to respond to additions of the other substrates for the reaction is in marked contrast to the behavior found for the natural nucleotide substrates and is tentatively attributed to the lack of a protein-mediated interaction between the nucleotide and tetrahydrofolate binding sites in the analog complex . The frequency dependence of solvent PRR in the presence of the various complexes allows an estimate of the correlation times for electron-nuclear dipolar interaction and thereby the extent of hydration of the bound Mn(II) among the various complexes..

J Biol Chem, 1975 Jan 10, 250(1), 254 - 60
Equilibrium and water proton relaxation rate enhancement properties of formyltetrahydrofolate synthetase-manganous ion-substrate complexes; Buttlaire DH et al.; Binding of Mn(pi)-nucleotide complexes to the enzyme formyltertrahydrofolate synthetase (EC 6.3.4.3) from Clostridium cylindrosporum has been examined in the presence and absence of other substrates by solvent proton relaxation mearurements . MnADP and MnATP form ternary complexes with the enzyme with highly enhanced proton relaxation rates for water . The enhancement parameters, epsilont, for the MnADP and MnATP ternary complexes are 19.8 and 12.5, respectively at 24.3 MHZ and 25 degrees . Titration curves with constant total concentrations of enzyme and Mn(pi) with variable nucleotide concentration are similar to those observed in similar titrations with the endp and MnATP are 175 muM and 64 muM, respectively at 25 degrees . Addition of tetrahydrofolate to solutions of the MnADP OR MnATP ternary complexes lowers the observed relaxation enhancement markedly . An analysis of titration curves with constant total concentrations of enzyme, Mn(pi), and nucleotide with variable tetrahydrofolate concentration gives the dissociation constant for tetrahydrofolate from the respective quaternary complexes . The affinity of the enzyme for tetrahydrofolate is increased 6-fold when MnADP is present at the active site whereas a 3-fold increase is observed with MnATP present . Furthermore, there is a 20-fold increase in the enzyme's affinity for tetrahydrofolate when both MnADP and the third substrate, formate, are present . The observed relaxation rate of water for solutions of the complex, enzyme-MnADP-tetrahydrofolate-formate, is deenhanced with respect to the rate observed for the simple aquo-Mn(pi) solution . Addition of nitrate to solutions of the above complex increases the affinity of the enzyme for tetrahydrofolate and MnADP by an additional factor of 5 and lowers the relaxation rate further to a value which approaches that for solutions of the enzyme and substrates which lack the paramagnetic cation.

Biochim Biophys Acta, 1975 Jan 6, 378(1), 44 - 53
Interaction of 4,5-dibromo-2,7-di-(acetatomercuri)-fluorescein with DNAs of different base composition; Dattagupta N et al.; The changes in absorption spectra in the visible region observed on adding different naturally occurring and synthetic DNA duplexes to solutions of 4,5-dibromo-2,7-di-(acetatomercuri)-fluorescein indicate that the mercurial reacts with polynucleotides of this type . The reaction is reversible as proved by adding excess of KCN which restores the original spectra of the free dye . The interaction is characterised also by quenching of the fluorescence of the dye and the induction of optical activity in it . The extent of these spectral effects depends strongly on the (A+T) content of the complexed DNA and decreased in the order: poly {d(A-T)}, Clostridium perfringens DAN, Escherichia coli DNA, Micrococcus luteus DNA and poly(dC) . From equilibrium-dialysis experiments the same order in affinity is obtained when these poly-nucleotides are at equilibrium with the same concentration of 4,5-dibromo-2,7-di-(acetatomercuri)-fluorescein . From the changes produced by different mercurials in the ORD spectra and viscosity of a DNA solution it has been concluded that 4,5-dibromo-2,7-di(acetatomercuri)-fluorescein does not cause any drastic alteration of the secondary structure of DNA.

Acta Biol Med Ger, 1975, 34(9), 1557 - 9
{Studies on human sera for antibodies against Clostridium butyricum strain Jean H 8}; Mehnert WH et al.; In 235 human sera, antibodies against Clostridium butyricum strain Jena H8 could be demonstrated in 33 patients (15%); only 189 sera (80%) reacted negatively, and in 5% (13 sera) a questionable reaction was obtained . The results are discussed, pointing out the necessity of preliminary titer determination if the clostridia-tumor phenomenon is to be utilized for early diagnosis of cancer.

Arch Geschwulstforsch, 1975, 45(2), 111 - 20
{Experiments concerning the enhancement of clostridial growth (Clostridium butyricum 1672 A McClung) by means of the anaerobic metabolism of Ehrlich ascites carcinoma cells of the mouse (author's transl)}; Negelein E et al.; Ehrlich ascites carcinoma cells stimulate the vegetative growth of Clostridium butyricum in a modified Parker medium under N2 atmosphere . The enhancement of clostridial growth depends on the number of tumour cells added . The Ehrlich ascites carcinoma cells keep the ordinary transplantability after anaerobic incubation with the vegetative forms of Clostridia . Further means for the manometric investigations of the tumour clostridium phenomenon are discussed.

Biochimie, 1975, 57(5), 603 - 8
{Purification and chemical study of a Collocalia glycoprotein}; Houdret N et al.; A glycoprotein was purified from the aqueous extract of "edible bird's nest" (Collocalia) using free flow preparative electrophoresis and represented the main fraction of Collocalia glycoproteins . This glycoprotein is homogeneous upon agarose electrophoresis and slightly polydisperse upon ultracentrifugation (S So 20w = 3,0) . The carbohydrate moiety contains galactose, mannose, glucosamine, galactosamine and sialic acid, which is completely released by Clostridium perfringens or Diplococcus pneumoniae neuraminidases and has the same chromatographic behaviour as N-acetyl-neuraminic acid . The peptide part of the glycoprotein is rich in serine, threonine and proline . About 40 p . cent of the hydroxyaminoacids are involved in carbohydrate-peptide linkages.

Zentralbl Bakteriol {Orig A}, 1975, 230(2), 241 - 5
{Studies on the culture of clostridium butyricum strain M 55 ATCC 13.732 . 4th Communication: The influence of peptone quality upon kininase activity (author's transl)}; Brantner H et al.; Studies of the oncolytic effect of Cl . butyricum strain M 55 resulted in the hypothesis of oncolysis to be an effect of a continuous production of kinin-decomposing enzymes . A decomposition of bradykinin will reduce capillary permeability and microcirculation in the tumour tissue, thus releasing and maintaining the process of necrotization . To achieve an optimal oncolytic effect of Cl . butyricum M 55, in addition to conditions already demanded in preceding papers, also an abundant production of kininase is required . Experience has shown peptone-containing media to be most suitable for culturing the M 55 strain, kininase formation being dependent upon peptone quality . The best yield of kinine-decomposing enzymes was achieved with the following peptones: peptone from Gelatine Merck, Tryptose Merck, Microbiotone Oxoid, Proteose Oxoid and Trypticase BBL . To obtain the greatest possible kininase production by the spores of Cl . butyricum M 55 applied for oncolytic therapy, the peptones mentioned are recommended for culture.

Zentralbl Bakteriol {Orig A}, 1975, 230(2), 237 - 40
Induction of chain formation in Clostridium sporogenes by suramin; Rafi Shaikh M et al.; Cl . sporogenes NCTC 532 was grown in serial doubling dilutions of Suramin from 1% to 0.125% w/v in BHI broth containing 1% glucose . After overnight incubation the culture grew into extremely long chains on transfer into suramin free medium reverted to normal morphology.

Scand J Gastroenterol, 1975, 10(1), 109 - 11
Clostridium septicemia following biliary surgery in a gastrectomized patient; Aukee S et al.; A 75-year-old woman was subjected to biliary surgery 38 years after partial gastrectomy for ulcer . There was a history of gallstones of 10 years duration, pentagastrin-resistant achylia, cholecystolithiasis and choledocholithiasis complicated by stenosis of papilla of vater, cholecystitis and pancreatitis . Peroperative cholangiography and biliary tract surgery were performed . On the third postoperative day heavy jaundice and hemolysis developed, leading to death of the patient . Culture of bile taken at operation revealed strains of Clostridium perfringens and Escherichia coli . Autopsy showed a picture of gas gangraena of the liver and Clostridium septicemia . The role of achylia, blind loop, and biliary obstruction in bile surgery is stressed.

Arch Geschwulstforsch, 1975, 45(1), 16 - 8
{A simple method for breeding major spore quantities of Clostridium butyricum to be used in experiments for tumour diagnostics and therapy (author's transl)}; Mehnert WH et al.; A simple method is described for breeding major spore quantities of Clostridium butyricum using a modification of Berger's iron bouillon . The method allows the breeding of cultures with comparatively high spore-forming rates which is essential for the use of such spores in experiments in the field of early tumour diagnostics and tumour therapy.

Int Arch Allergy Appl Immunol, 1975, 48(4), 495 - 504
On the question of permeability of the blood-brain barrier to botulinum toxin; Boroff DA et al.; The clinical symptoms of botulinum intoxication suggest that besides the involvement of the peripheral nervous system, the central nervous system is also affected . Studies were undertaken to determine whether pure toxin of Clostridium botulinum type A could be demonstrated in the brains of poisoned mice . With the aid of autoradiography of the toxin marked with 125-I and indirect fluorescent labeling it was possible to show the presence of the toxin in the parenchyma of the brain.

Avian Dis, 1975 Jan-Mar, 19(1), 192 - 5
An outbreak of type-C botulism in three-weeek-old broilers; Page RK et al.; Botulism was diagnosed in 3-week-old broilers from clinical signs, absence of postmortem and histopathological lesions, and demonstration of toxin in the serum of comatose birds . Passive immunization of mice with Clostridium botulinum type-C antitoxin protected against a challenge with serum from comatose birds containing the Cl . botulinum toxin . Total mortality for the grow-out period exceeded 27% and was not altered by water medication with penicillin . Bacitracin at 100 g per ton reduced mortality to 5-7 birds per day.

Acta Microbiol Acad Sci Hung, 1975, 22(4), 397 - 402
{Application of microbial enzymes in studies of steroid metabolism (author's transl)}; Schlegel J et al.; In vitro models are necessary for studying the biotransformation of new steroid drugs and to determine the structure of the metabolites and their biological activity . For that reason microorganisms and their enzymes were used to investigate the anabolic steroid Oral-Turinabol (4-chloro-17alpha-methyl-17beta-hydroxy-1,4-androstadiene-3-one) . Clostridium paraputrificum transformed Oral-Turinabol into the hydrogenation products 4beta-chloro-17alpha-methyl-17beta-hydroxy-5beta-1-androstene-3-one (I) and 4beta-chloro-17alpha-methyl-5beta-1-androstene-3alpha,17beta-biol (II); Rhodotorula glutinis to 4alpha-chloro-17alpha-methyl-5alpha-1-androstene-3beta,17beta-hydroxy-5alpha-1-androstene-3-one (III) and 4alpha-chloro-17alpha-methyl-5alpha-1-androstene-3beta,17beta-diol (IV) . The hydroxylation products 6beta-hydroxy- (V), 7beta-hydroxy- (VI), 15alpha- (VII) and 15beta-hydroxy-Oral-Turinabol (VIII) resulted from Absidia glauca and Aspergillus flavus . The metabolites II-V were isolated until now from mammals.

Vet Med Nauki, 1975, 12(6), 84 - 8
{Comparative study of the protective qualities of some protease inhibitors in the drying of Cl . oedematiens type B toxin}; Peichev B; A comparative study was carried out on the lethal properties of dry toxins of Clostridium oedematiens Type B obtained by means of salting cultural Seitz-filtrates, containing or lacking protease inhibitors, being dried for 24 and 48 hours at 37 degrees C . It was found that when ammonium sulfate was used for salting, the dry toxins from such filtrates, containing 0.0001 gram% specific soya inhibitor, showed 2 to 300 times higher toxicity than those containing no inhibitor . The addition of 3 per cent normal horse serum to the cultural filtrates of Clostridium oedematiens Type B favoured the production of dry toxins of higher lethal properties . The decrease in the lethal properties of a dry toxin without inhibitor was found to be in a positive correlation with the drying term and the residual moisture of the toxin . When a protease inhibitor was employed the drying term had no effect on the activity of the dry toxins.

Arch Microbiol, 1975, 102(2), 145 - 9
The metabolism of pyrimidines by proteolytic clostridia; Hilton MG; Uracil was used by growing cultures of Clostridium sporogenes, and by proteolytic strains of C . botulinum types A and B . Uracil was not used by C . bifermentans; C . botulinum, type B (non-proteolytic); C . botulinum, type F (non-proteolytic); C . botulinum, type E; C . butyricum; C . cochlearium; C . difficile; C . histolyticum; C . oedematiens, type A; C . paraputrificum; C . scatologenes; C . specticum; C . sordellii; C . sticklandii; C . tertium; C . tetani; C . tetanomorphum; C . welchii, types A, B, C, E and 4 untyped strains . The growth of C . sporogenes was not increased by uracil; it was reduced to dihydrouracil . Experiments with washed cells of C . sporogenes showed that the uracil-reducing system was inducible . Washed cell suspensions incubated under hydrogen with uracil, thymine, iso-barbituric acid, 5-amino uracil and cytosine consumed 1 mole H2/mole pyrimidine . The reduction product of cytosine was dihydrouracil indicating that it was deaminated before reduction . The reduction products of the remaining pyrimidines were the corresponding dihydro derivatives . Extracts of C . sporogenes reduced uracil in the presence of NADPH2 but not NADH2.

J Bacteriol, 1975 Jan, 121(1), 184 - 91
Factor 420-dependent tyridine nucleotide-linked hydrogenase system of Methanobacterium ruminantium; Tzeng SF et al.; Methanobacterium ruminantium was shown to possess a nicotinamide adenine dinucleotide phosphate (NADP)-linked factor 420 (F420)-dependent hydrogenase system . This system was also shown to be present in Methanobacterium strain MOH . The hydrogenase system of M . ruminantium also links directly to F420, flavin adenine dinucleotide (FAD), flavin mononucleotide (FMN), methyl viologen, and Fe-3 plus . It has a pH optimum of about 8 and an apparent Km for F420 of about 5 x 10-6 M at pH 8 when NADP is the electron acceptor . The F420-NADP oxidoreductase activity is inactive toward nicotinamide adenine dinucleotide (nad) and no NADPH:NAD or FADH2(FMNH2):NAD transhydrogenase system was detected . Neither crude ferredoxin nor boiled crude extract of Clostridium pasteuranum could replace F420 in the NADP-linked hydrogenase reaction of M . ruminantium . Also, neitther F420 nor a curde "ferredoxin" fraction from M . ruminantium extracts could substitute for ferredoxin in the pyruvate-ferredoxin oxidoreductase reaction of C . pasteurianum.

Scand J Infect Dis, 1975, 7(2), 135 - 9
Regression line analysis for five antibiotics with strains of Clostridium species; Dornbusch K et al.; The susceptibility of 90 strains of clostridia to 5 antibacterials was determined by the agar plate dilution method and the disc diffusion test . The correlations between minimum inhibitory concentration and inhibition zone were quite acceptable for lincomycin, clindamycin, tetracycline and doxycycline to calculate regression lines by the least square method . The strains showed comparatively wide ranges of susceptibility . For erythromycin, no regression line could be calculated, since all the strains tested were susceptible over a narrow range of concentrations.

Scand J Infect Dis, 1975, 7(2), 127 - 34
Antibiotic susceptibility of Clostridium species isolated from human infections; Dornbusch K et al.; In antibacterial susceptibility testing by the agar plate dilution method with 252 strains of Clostridium perfringens, 22 strains of C . novyi type A and 7 of C . novyi type B, with 21 strains of C . bifermentans, 12 strains of C . sordellii and 18 of C . sporogenes, the majority of the strains were inhibited by penicillin G, cephalothin, chloramphenicol, erythromycin, tinidazole and metronidazole . Resistance to lincomycin and clindamycin was noticed among strains of all clostridia species as well as to tetracycline and doxycycline among strains of C . perfringens.

Zentralbl Bakteriol {Orig A}, 1975, 230(2), 246 - 51
Outbursting wedges of the colonies of Clostridium welchii containing chain forming mutants after ultra violet irradiation; Rafi Shaikh M et al.; Cl . welchii NCTC 6785 was irradiated with UV light . The 40 minutes irradiated cells after 3-8 days ageing at room temperature (22-25 degrees C) and 48 hours initial incubation at 37 degrees C produced hairy outbursting wedges . These wedges gave rise to the colonies composed of chain forming mutants, which on transfer into the fluid medium grew into a pure chain forming culture.

Appl Microbiol, 1975 Jan, 29(1), 1 - 6
Minimal growth requirements for Clostridium perfringens and isolation of auxotrophic mutants; Sebald M et al.; The minimal growth requirements for two strains of Clostridium perfringens were defined, and both synthetic and semisynthetic plating media were developed . Plate counts of the wild-type strains on both of these minimal media were equivalent to those on complex media . A number of auxotrophic mutants of each strain were isolated, and their phenotypes were defined.

J Infect Dis, 1975 Jan, 131(1), 58 - 63
Classification of enterotoxins on the basis of activity in cell culture; Keusch GT et al.; Two cell culture systems were used in a study of the biological properties of several bacterial enterotoxins in vitro . By means of one model, in which HeLa cell monolayers were used, cytotoxic effects, interms of detachment of cells from a glass surface due to cell death, were assayed . By means of the second model, activation of the adenyl cyclase-cyclic adenosine 3', 5'-monophosphate (AMP) system, in terms of increased steroidogenesis by Y-1 adrenal cells (an effect which we have termed cytotonic), was assayed . None of the four toxins manifested both properties . Rather there was a clear segregation into two cytotoxic enterotoxins (Shigella dysenteriae type 1 and Clostridium perfingens) and two cytotonic products (Vibrio cholerae and Escherichia coli) . These data raise the possibility that some enterotoxins may not interact with the adenyl cyclase-cyclic AMP system; this possibility has also been suggested by studies of the toxin of S . dysenteriae type 1 in the rabbit small bowel . These cell culture systems may therefore serve as convenient models for the study of the mechanism of action of both classes of enterotoxin.

Acta Med Iran, 1975, 18(3-4), 111 - 28
Clostridium difficile; Modaber I; Seventy-five meconium samples were examined for the presence of Cl . difficile; 3 strains were isolated . Additionally 45 laboratory animal faeces specimens were tested for the same purpose, a further 2 cases were isolated . These five suspicious strains were identified as Cl . difficle according to the tests mentioned in the previous paragraphs . The organisms isolated here showed the same characteristics as five of the strains received and also as the organisms isolated from the inoculated animals with the crude cultures of Cl . difficile . These organisms were variable in size, roughly 2-9 XO.3-0 . 8u, Gram positive rods, motile, capsulated, flagellated, most probably peritrichous, possessing non-bulging spores located terminally or subterminally, free spores were rarely detectable . Cell arrangements: singly or in pairs and occasionally in short chains . On longer incubation the organisms slightly shifted to become Gram variable and longer in size . Colonies on ordinary agar and solid blood agar appeared to be punctiform and rough . On the other hand the colony appearance on the rest of the solid media which are mentioned previously are as follows: 1-3 mm in diameter, greenish, smooth, non-haemolytic, entire some showing slight irregularities of their edges . Colonies slightly raised, butyrous and semi opaque to opaque . This organism does not liquify the serum of Loeffler medium and also does not cause any changes of this medium . The metachromatic granules are readily seen by Albert's staining . Neither proteolytic nor lipolytic activities are possessed by this organism . Sensitivity to antibiotics showed the same pattern as mentioned about the strains received.

J Membr Biol, 1975, 23(3-4), 293 - 204
Phospholipases . III . Effects of ionic surfactants on the phospholipase-catalyzed hydrolysis of unsonicated egg lecithin liposomes; Goldhammer AR et al.; Apparent values of Km and Vmax have been measured for catalysis of hydrolysis of unsonicated egg lecithin liposomes, activated through addition of 0.4 M n-hexanol, by phospholipases A2 from bee and snake venoms and by phospholipase C from Clostridium welchii as a function of the concentration of three surfactants: hexadecylamine, hexadecyltrimethylammonium bromide, and dihexadecyl phosphate . For all three enzymes, values of Km and Vmax show little or no dependence on the concentration of these ionic surfactants, demonstrating that the liposomal surface charge is not a crucial factor in determining susceptibility to phospholipase-catalyzed hydrolysis.

Jpn J Med Sci Biol, 1974 Dec, 27(6), 285 - 96
Isolation and characterization of an esterase-active enzyme from pronase with special reference to activation of Clostridium botulinum type E progenitor toxin; Miura T; (1) By sequential chromatography on CM-Sephadex, SP-Sephadex and Sephadex G-100, an esterase-active enzyme was isolated from a commercial preparation of pronase, (2) The purity of the isolated enzyme was proven by disc electrophoresis and sedimentation analysis . (3) The molecular weight of the esterase-active enzyme was approximately 16,000 . The sedimentation coefficient was 1.85 S . The N- and C-terminal amino acids were asparagine and valine, respectively . (4) The enzyme was similar to that of bovine trypsin in the substrate specificity . It appeared to be the same as the trypsin-like enzyme isolated from pronase by Wahyby (1968) . However, the relative rates of hydrolyses of different substrates by the enzyme differed from those of bovine trypsin . (5) The enzyme activated Clostridium botulinum type E progenitor toxin . Even a higher toxicity was resulted by treating the progenitor toxin with this enzyme than with bovine trypsin . (6) The results support the hypothesis that activation of type E progenitor toxin involves hydrolysis of a lysine ester bond.

J Lipid Res, 1974 Sep, 15(5), 500 - 7
Phospholipids of Clostridium butyricum . V . Effects of growth temperature on fatty acid, alk-1-enyl ether group, and phospholipid composition; Khuller GK et al.; Many anaerobic bacteria have a high proportion of 1-alk-1'-enyl ethers (plasmalogens) among their phospholipids . We have examined the effects of growth temperature on the phospholipid, fatty acid, and alk-1-enyl group compositions of Clostridium butyricum . When the growth temperature was decreased from 37 degrees C to 25 degrees C, the proportion of glycerol phosphoglycerides (the sum of phosphatidylglycerol and the corresponding plasmalogen) increased at the expense of the ethanolamine and N-methylethanolamine phosphoglycerides . Analysis of the proportion of these lipids present in the diacyl and 1-alk-1'-enyl-2-acyl forms has shown a substantial increase in the plasmalogen form of the glycerol phosphoglycerides and a decrease in the plasmalogen forms of the ethanolamine and N-methylethanolamine phosphoglycerides . An analysis of the fatty acids and alk-1-enyl groups isolated from the total phospholipids of cells grown at 25 degrees C, 30 degrees C, and 37 degrees C has shown a general increase in the proportions of unsaturated and cyclopropane hydrocarbon chains at lower growth temperatures . When the temperature was lowered from 37 degrees C to 25 degrees C, the fatty acids had progressively more unsaturated and cyclopropane chains and fewer saturated chains . The alk-1-enyl groups, in particular those from the ethanolamine and N-methylethanolamine plasmalogens, were more saturated at 30 degrees C than at 37 degrees C . When the growth temperature was lowered to 25 degrees C, there was little further change in the degree of unsaturation of the alk-1-enyl groups.

J Lipid Res, 1971 Mar, 12(2), 214 - 20
Phospholipids of Clostridium butyricum . IV . Analysis of the positional isomers of monounsaturated and cyclopropane fatty acids and alk-1'-enyl ethers by capillary column chromatography; Goldfine H et al.; The positional isomers of the cyclopropane fatty acids of Clostridium butyricum phospholipids have been analyzed by capillary column gas-liquid chromatography . Greater than 95% of the methylenehexadecanoic acids was the 9,10 isomer . On the other hand, 60-70% of the hexadecenoic acid precursors was the Delta(7) isomer, and the remainder was the Delta(9) isomer . Of the methyleneoctadecanoic acids 75-80% was the 11,12 isomer, with the remainder being the 9,10 isomer . There were approximately equal amounts of the Delta(9)- and Delta(11)-octadecenoic acids in the phospholipids . This study reveals a surprisingly strong specificity of the cyclopropane synthetase for the (n-7) series of monoenoic fatty acids . An analysis by capillary column chromatography of the monoenoic and cyclopropane aldehyde dimethylacetals derived from the plasmalogens (1-alk-1'-enyl-2-acyl-glycero-phosphatides) of C . butyricum revealed the presence of the same positional isomeric mixtures of the 16- and 18-carbon monoenoic residues in approximately the same ratios as were found in the fatty acids . In the formation of the cyclopropane alk-1'-enyl ethers there was also specificity for the (n-7) series, but it was not as strong as that seen in the fatty acids . The ratio of the 7,8 isomer to the 9,10 isomer was higher in the methyl-enehexadecanals than in the corresponding fatty acids . This paper extends the use of Golay capillary columns to the analysis of the positional isomers of plasmalogen aldehydes as their dimethylacetal derivatives.

Clin Obstet Gynecol, 1970 Jun, 13(2), 291 - 304
Septic abortion and septic shock; Santamarina BA et al.; PIP: Septic shock in obstetric cases is described, and patient management is discussed . Cases of septic abortion complicated by septic shock differ from those resulting from bacterial shock . During the 10 years prior to 1968, Boston City Hospital had a yearly average of 550-600 abortions, and about 26% were septic . Septic shock developed in 3.8% of patients . During 1968 and 1969, the number of abortions dropped to 491 and 461, respectively, with 107 septic for 1968 and 73 for 1969; during these years, only 4 cases had septic shock, but these resulted in 2 maternal deaths . Both of these cases had been mistreated due to inappropriate diagnosis . The pathophysiology and management of these cases are described . Most septic abortions complicated by septic shock result from gram-negative organisms which produce endotoxin; in addition, 2 gram-positive organisms, Clostridium welchii and C . perfringens, are frequently responsible for septic shock . Often, illegal abortion was the cause for septic abortion and shock, but it can also occur with prolonged ruptured membranes and chorioamnionitis in the last trimester . These patients presented with a variety of clinical symptoms, including septic shock . They required intensive therapy; about 95% of all septic abortions can be controlled and respond favorably to adequate antibiotic therapy and evacuation of uterine contents . If the patient fails to respond to this form of therapy, septic shock is frequently a complication . Under these circumstances, immediate surgical intervention, in the form of total abdominal hysterectomy and bilateral salpingo-oophorectomy, is performed, and the vaginal cuff is left open to allow for adequate drainage . Without such treatment, mortality rates increase to 60% . The use of vasopressors if central venous pressure is very low is recommended by this hospital group, Isuprel in particular . Exchange transfusion is advocated to combat hemolysis .

Am J Obstet Gynecol, 1966 Nov 1, 96(5), 633 - 44
Clinical management of septic abortion complicated by hypotension; Douglas GW et al.; PIP: A mortality of 22% is reported for 50 cases of septic abortion complicated by hypotension . Patients were treated by a variety of medical managements, and 44 underwent surgery . 38% were subjected to hysterectomy based on the following indications: 1) unresponsive septic, hypotension, large uterus; 2) intrauterine douche with necrosis; and 3) anuria . Experience with this series indicated the following procedural principles: 1) hourly recording of blood pressure and urinary output; 2) gram-stained smears from cervix; 3) infusion of intravenous fluids and antibiotics geared to combat gram-negative organisms and Clostridium welchii (vasopressor may be required for patients in deep shock); and 4) surgical treatment to remove the source of infection and toxicity . Rationale for clinical management is discussed .

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