Clostridia

Microbiol Immunol, 1979, 23(6), 443 - 54
Isolation and partial characterization of exosporium from spores of a highly sporogenic mutant of Clostridium botulinum type A; Takumi K et al.; Homogeneous fragments of exosporium were isolated and purified from mature spores of a highly sporogenic mutant derived from Clostridium botulinum type A strain 190L . The exosporium was composed of three lamellae and showed a hexagonal array when negatively stained . The hexagonal array of isolated exosporium was resistant to sodium dodecyl sulfate, urea, dithiothreitol, and proteolytic enzymes such as trypsin, pronase, and nagarse, except for pepsin . The hexagonal array was partially disintegrated with 5 M guanidine-HCl and almost completely disrupted with 8 M urea in combination with 1% mercaptoethanol under alkaline conditions . The purified exosporium fraction was composed mainly of protein (69.1%) and lipids (13.8%) . A small amount of amino sugars (2.5%) was present, but neutral sugars could not be detected . The exosporium protein had a predominantly acidic amino acid composition accompanied by low levels of cystine, methionine, and histidine.

Am J Clin Nutr, 1979 Jan, 32(1), 251 - 7
Diarrhea and colitis associated with antimicrobial therapy in man and animals; George WL et al.; Antimicrobial agent-induced ileocecitis of laboratory animals and colitis of man share common features . The significance of a newly described toxin in these two entities, the apparent source of the toxin (Clostridium difficile) and characteristics of the toxin are reviewed . Methods of toxin detection, isolation and rapid identification of C . difficile, and possible modes of therapy for antimicrobial agent-associated colitis of man are discussed.

Am J Clin Nutr, 1979 Jan, 32(1), 113 - 8
Introduction to mechanisms of association of indigenous microbes; Savage DC; Indigenous microorganisms of numerous types associate with epithelial surfaces in the gastrointestinal tracts of mammals and birds . Some of the microbial types, e.g., Lactobacillus sp . in the stomachs of laboratory rodents, adhere to a surface without altering it ultrastructurely . In such cases, the adherence is mediated undoubtedly by macromolecules on the bacterial surfaces, possibly polysaccharides in most cases, interacting in specific ways with receptor macromolecules on the epithelial surface . Other microbial types that associate with epithelial surfaces without altering them ultrastructurally, e.g., Clostridium 109-2 in the mouse large bowel, may adhere to the surface only weakly or not at all, and maintain the association because they are motile and attracted to the epithelium by chemotactic substances . Microbial types that alter the ultrastructure of the epithelial cells to which they attach interact intimately with the membranes of the epithelial cells . In such cases, the microbes have specialized segments or ends for adhering to the membranes, and probably elaborate systems for stabilizing the membranes and cytoplasm at the site in the epithelial cell to which they attach . Some such organisms may have evolved unique reproductive mechanisms to maintain their populations on the epithelial surface.

Rev Infect Dis, 1979 Jan-Feb, 1(1), 218 - 23
In vitro activity of cefoxitin and parenterally administered cephalosporins against anaerobic bacteria; Sutter VL et al.; The in vitro activity of cefoxitin against 221 recent isolates of anaerobic bacteria was compared with that of cefamandole and cefuroxime by the agar dilution technique . At achievable serum levels of 32 micrograms/ml, all three drugs were active against most groups of anaerobic bacteria tested . Cefoxitin was active against most strains of the Bacteroides fragilis group, whereas cefamandole and cefuroxime were relatively ineffective . Cefoxitin was least active against Clostridium species other than Clostridium perfringens, whereas cefamandole was very active and cefuroxime moderately active . A review of recent in vitro studies with cefoxitin and parenterally administered cephalosporins indicates that cefoxitin is the most active compound against the B . fragilis group and is least active against Clostridium species other than C . perfringens.

Arch Exp Veterinarmed, 1979, 33(4), 621 - 37
{Studies of necrotizing enteritis of suckling piglets (Clostridium perfringens type C enterotoxemia) in industrialized sow breeding units . 5 . Control of the disease}; Kohler B et al.; Recent methods used and experience obtained in the control of necrotising enteritis are reported in this paper, with reference being made to both the pathogenesis and epizootiology of the disease . Two inoculations of the sows, using "Enterotoxamievakzine Dessau bivalent" five and three weeks before parturition, have worked well for prophylaxis . Oral treatment was applied to nursed piglets, using 40,000 I.U . of "Aviapen" and "V-Tablopen" penicillin per animal and day over periods between two and four days, helped to minimise piglet loss, particularly in the period between a fresh outbreak and full effectiveness of immunoprophylactic action . Such treatment was conducted metaphylactically and therapeutically . The first metaphylactic treatment was given within 24 hours from parturition . Combination of mother animal vaccination with the above therapeutic use of those two penicillin preparations worked extremely well in enzootically contaminated stocks and proved to be the most effective approach, for the time being, to controlling necrotising enteritis of nursed piglets . Yet, all those control measures failed to bring about full stock sanitation on industrialised units . Sow trading was not permitted until at least four weeks had elapsed from full effectiveness of mother animal vaccination, with the view to reducing the proliferation of Clostridium perfringens Type C via sales of breeding animals . All sows were given two "Enterotoxamievakzine Dessau bivalent" vaccinations, prior to sale . The animals were sold only to smaller farms (less than 500 sows for breeding) with concentional keeping patterns which were kept under constant diagnostic supervision . Neomycin, oxytetracycline, chloramphenicol, and other antibiotics against which Clostridium perfringens was resistant or in a position to assume resistance were used on endangered stocks only in conjunction with penicillin or not at all . This programme of control has proved to be efficient through a period of more than three years.

Arch Exp Veterinarmed, 1979, 33(4), 595 - 619
{Studies of necrotizing enteritis of suckling piglets (Clostridium perfringens type C enterotoxemia) in industrialized sow breeding units . 4 . Epizootiology}; Kohler B et al.; Necrotising enteritis had been the cause of death of 4.9 per cent in 5,177 nursed piglets, which was established by pathological examination . The number of piglets, in that context, which had come from industrialised sow breeding units was equivalent to 92 per cent . The nursed piglet held the third position, next to smaller ruminants (19.4 per cent) and fowl (6.0 per cent), with regard to the occurrence of Clostridium perfringens enterotoxemia or necrotising enteritis in 112,218 animals which were pathologically examined after death . Necrotising enteritis so far has been rare in the GDR . No regional accumulation has been observed . Several outbreaks on industrialised sow breeding units actually remained stationary . The occurrence of the disease may be favoured by a number of factors which are conducive to accumulation of Clostridium perfringens Type C in a given stock . Group keeping of pregnant sows, simultaneous farrowing of larger groups of sows, group treatment of nursed piglets, using neomycin, chloramphenicol, oxytetracycline, and other antibiotics to which Clostridium perfringens is primarily resistant or has acquired resistance in the course of time are some of those contributive factors . Transmission of Clostridium perfringens Type C through feedstuff is possible, though it would lead to a real outbreak only by high intensity of the contamination, and it played a minor role in proliferation of the disease . 3479 Clostridium perfringens strains were isolated from 9,481 animals, both clinically intact and after death, with 30 species being included . Type classification revealed 2454 strains of Type A (70 per cent), 204 of Type D (5.88 per cent), 164 of Type C (four per cent), and 48 of Type B (1.34 per cent) . There were 688 atoxic strains (17 per cent) . Swine is the major carrier of Clostridium perfringens Type C, with 87 per cent of all Clostridium perfringens Type C strains having been isolated from swine . Swine was followed by fowl (four per cent), sheep (four per cent), cattle, rabbit, and dog (1.27 per cent each) . Clostridium perfringens Type C was obtained from the faeces of clinically intact sows in seven instances, including two cases with sows (0.46 per cent) from farms with no previous record of necrotising enteritis.

Microbios, 1979, 25(100), 85 - 91
Sensitivity of chemically treated spores of Clostridium perfringens type A to an initiation protein; Franceschini TJ et al.; Extraction of Clostridium perfringens type A spores with dithiothreitol (DTT), DTT plus sodium dodecyl sulphate (DTT-SDS), urea-mercaptoethanol (UME), or alkali, solubilized from 18.6 to 46.5 of the total dry weight of spores . The initiation of germination and lysis of such treated spores with lysozyme and an initiation protein (IP) from the culture supernatant fluid of sporulating cells of C . perfringens was studied under various conditions . The ability of lysozyme and the crude IP to induce germination and lysis of extracted spores was concentration dependent up to 0.5 microgram/ml and 5.6 mg/ml respectively . IP showed an optimum of activity between pH 7 and 8 for DTT-SDS and DTT extracted spores, and between pH 6 and 9 for UME extracted spores . The optimum temperature of activity for IP was 55 degrees C . Dissimilarities in the extent to which lysozyme and the IP initiated germination and lysis of spores extracted by various methods may have been a reflection of the differences in amounts of protein solubilized by each treatment.

J Biol Stand, 1979, 7(4), 373 - 81
Development, preparation and safety testing of a Clostridium welchii type C toxoid . II . Laboratory evaluation of potency; Knight PA et al.; The results obtained with four laboratory tests on four candidate formulations of Clostridium welchii type C vaccine for use in man have been compared with clinical responses to the same vaccines . Quantal response assays in mice appeared to reflect the ranking of the four vaccines in human subjects better than did the guinea pig tests . They also enabled the potency of the vaccine preparations to be related to an existing International Reference Preparation . Mouse assays in which the animals received two spaced doses of vaccine prior to challenge yielded marginally more satisfactory results in terms of precision and reflection of human responses than did assays involving a single dose of vaccine.

J Biol Stand, 1979, 7(4), 315 - 23
Development, preparation and safety testing of a Clostridium welchii type C toxoid . I: preliminary observations in man in Papua New Guinea; Walker PD et al.; The reactogenicity and immunogenicity of various strengths of plain and adsorbed Cl . welchii type C toxoid have been evaluated in laboratory tests and in man in Papua New Guinea . The greater antigenicity and acceptable clinical reactivity of a vaccine containing 50 total combining power units per 0.5 mm of adsorbed toxoid resulted in its selection for further field studies.

Arch Exp Veterinarmed, 1979, 33(2), 313 - 33
{Studies of necrotizing enteritis of suckling piglets (Cl . perfringens typc C enterotoxemia) in industrialized sow breeding units . 3 . Experimental reproduction of the disease}; Kohler B et al.; Experimental reproduction of necrotising enteritis of sucking pigs was successfully achieved by using both Clostridium perfringens Type C strains, which had been isolated from sucking pigs with necrotising enteritis, and Type C strain 3628 of A.T.C.C . (sub-type C1) . The lethal dose for sucking pigs was between 20 X 10(6) and 12 X 10(7) pathogens per animal . The disease could not even be induced by repeated application of no-bacterial toxin of Cl . perfringens Type C nor by administration of Cl . perfringens Type A strains which had been cultured from broilers with necrotising enteritis . Necrotising enteritis was found to develop in two phases in sucking pigs . First, the pathogen will deposit to the villous epithelium and then penetrate the superficial strata of the mocous membrane . In the second phase, the villous structure will be destroyed by the lethal, haemolysing, and necrotising toxins of Cl . perfringens . The role played by individual toxin fractions is discussed together with the importance of humoral and localised infection defence . Sucking pigs may be sufficiently protected against infection based on single or ten-fold lethal infectious dosage by two vaccinations of the mother animal, five and three weeks prior to parturition, using "Enterotoxaemia Vaccine Dessau bivalent" . Infection then would not occur unless a hundredfold lethal dose was applied . Characteristics include diarrhoea, apathy, exhaustion, and death.

Microbiol Immunol, 1979, 23(4), 213 - 21
Pathogenesis of Hobbs' heat-sensitive spore forming Clostridium perfringens type A strain; Chakrabarty AK et al.; Food poisoning in man due to heat-sensitive strains of Cl . perfringens type A appeared to be mediated through enterotoxin synthesized in vivo during sporulation . A minimum of 2.0 X 10(5) vegetative cells suspended in sporulation medium was sufficient to elicit gut-loop response in rabbits . The functional disturbance in the gut as well as the structural changes were progressive and proportional to the size of the inoculum up to a dose limit of 2.0 X 10(7) vegetative cells and beyond this the changes remained steady.

Can J Comp Med, 1979 Jan, 43(1), 98 - 101
Identification of enterotoxigenic Clostridium perfringens type A in mixed cultures; Niilo L; Three known enterotoxigenic Clostridium perfringens type A strains were mixed in various combinations with three nonenterotoxigenic strains and three lots of animal intestinal contents . They were grown as mixed cultures and tested for the presence of enterotoxin by the fluorescent antibody, reversed passive hemagglutination and immunodiffusion techniques . The fluorescent antibody and reversed passive hemagglutination tests detected enterotoxin in all 16 cultures prepared but the immunodiffusion test failed on two cultures . Attempts to reisolate the enterotoxigenic strains from the mixed cultures was successful in 12 cases when two of five isolated colonies were selected.

Zh Mikrobiol Epidemiol Immunobiol, 1979 Jan, (1), 43 - 6
{Electron microscopic study of several features of spore formation in type B Clostridium perfringens}; Shakhbanov AA; The authors carried out electron microscopy of the thin sections of Cl . perfringens, type B (strain No . 89) . Material of middle electron density was revealed on the cell wall surface from the first hours of the culture growing; the cytoplasm displayed both rod-like incorporations with transverse striations, and phage particles . Different spore formation disturbances were revealed in the strain under study . In the majority of cells spore formation was blocked at the III--V stage . Besides, there were pseudospores, whereas mature spores were rarely encountered, and even those which did occur, were at the stage of growing.

Appl Environ Microbiol, 1979 Jan, 37(1), 55 - 66
Membrane filter enumeration method for Clostridium perfringens; Bisson JW et al.; A membrane filter procedure has been developed for the rapid quantitation of C . perfringens in the aquatic environment . Background growth is inhibited by the use of D-cycloserine, polymyxin B sulfate, and incubation at 45 degrees C . Differential characteristics include the fermentation of sucrose, production of acid phosphatase, and the absence of beta-D-glucosidase activity . The medium is prepared as follows (in grams per 100 ml of distilled water): tryptose, 3.0; yeast extract, 2.0; sucrose, 0.5; L-cysteine, 0.1; MgSO4 . 7H2O, 0.01; bromocresol purple, 0.004; and agar, 1.5 . The ingredients are dissolved, and the pH is adjusted to 7.6 . After autoclaving at 121 degrees C for 15 min, the medium is allowed to cool at 50 degrees C, and the following are added per 100 ml: D-cycloserine, 40 mg; polymyxin B sulfate, 2.5 mg; indoxyl-beta-D-glucoside, 60 mg; 2.0 ml of a filter-sterilized 0.5% phenolpthalein diphosphate solution; and 0.2 ml of a filter-sterilized 4.5% FeCl3.6H2O solution . Enumeration of C . perfringens in a water sample is completed within 18 to 24 h . The verification of typical colonies was 93% . The average recovery from peptone-water spore suspensions of five strains was 79%, and that from filter-sterilized seawater suspensions was 90% . The precision of the method was approximately equal to that expected from random error alone . Confirmed recoveries of C . perfringens from water and sewage samples generally were greater than those by the Bonde pour tube method.

Am J Clin Nutr, 1979 Jan, 32(1), 210 - 8
The molecular mode of action of Clostridium perfringens enterotoxin; McDonel JL; While certain strains of Clostridium perfringens have been associated with food poisoning outbreaks for the past 30 years, it has been only during the past 10 years that progress has been made in describing the disease process . And only within the past 5 years has meaningful progress been made in understanding the mechanism by which the disease is caused . Early observations, that the protein enterotoxin can cause erythema, increase capillary permeability, and exhibit parasympathomimetic properties, have been greatly added to in more recent studies . It is now believed tht the enterotoxin can alter intestinal transport of fluid, ions, and glucose, cause tissue damage in the gut and inhibit metabolic processes in intestinal tissue . Furthermore, the enterotoxin is thought to act very quickly (in a matter of minutes, compared to hours for other known enteropathogenic factors) and to affect basic function (macromolecular synthesis) and structure (membrane damage to microvillus brush borders) of individual cells . These findings have opened up many new questions that hopefully, when answered, will further the understanding of how this enterotoxin acts, as well as other enterotoxins being studied today.

Microbios, 1979, 24(97-98), 151 - 7
The formation of serine from glycine and formaldehyde by cell free extracts of Clostridium acidi-urici; Hougland AE et al.; Cells of Clostridium acidi-urici which were grown in a medium containing uric acid were harvested, disrupted by sonication and centrifuged . After centrifugation the supernatant which served as the cell free extract was used to study the synthesis of serine from 2-14C glycine and formaldehyde . Serine was isolated from the reaction mixture by column chromatography . After identification by paper chromatography, serine was degraded carbon by carbon to locate the position of the labelled carbon . Radioactivity was confined almost exclusively to the alpha carbon of serine which was derived from the alpha carbon of glycine . Formaldehyde, therefore, binds at the alpha carbon of glycine to form serine . Conversion of serine to pyruvate was prevented by adding EDTA to the reaction mixture.

Infect Immun, 1979 Jan, 23(1), 168 - 74
Microbial interference and colonization of the murine gastrointestinal tract by Listeria monocytogenes; Zachar Z et al.; Two strains of Listeria monocytogenes, one that formed smooth colonies on agar surfaces and a varient of it that formed rough colonies, colonized the gastrointestinal tracts of germfree mice . Within 24 h after mice were inoculated orally with about 100 bacteria, the population levels per gram (wet weight) of tissue of both strains were 10(5) to 10(7) in the stomach and ileum and 10(8) to 10(9) in the cecum and colon, respectively . As detected in Gram-stained histological sections, in such gnotobiotes, the bacteria colonized the lumen in all areas of the tract and much of the mucus layer on the epithelial surface in the proximal colon . The strain that formed smooth colonies did not colonize the tracts of specific-pathogen-free mice, but did colonize, to the same levels as in germfree mice, the stomachs and bowels of ex-germfree mice previously associated with two members of the indigenous flora (Bacteroides and Clostridium) . In the latter animals, however, the listeria did not form layers on the colonic epithelium as efficiently as they did in monoassociated gnotobiotes.

J Wildl Dis, 1979 Jan, 15(1), 25 - 31
Techniques for evaluating humoral and cell-mediated immunity in mule deer fawns (Odocoileus hemionus); Trindle BD et al.; Twenty mule deer fawns (Odocoileus hemionus) were removed from their dams 48 h after birth, and hand-reared . Methods for monitoring their immune capability are described . Passive humoral immunity was determined by serum protein electrophoresis . Active humoral immunity following Clostridium toxoid vaccination was determined by immunodiffusion . Cell-mediated immunity was assayed using contact sensitization to 1-nitro, 2,4-dichlorobenzene (DNCB).

Zentralbl Bakteriol {Orig A}, 1979, 245(3), 332 - 44
{Studies of the purification of the exotoxin of Bacillus cereus (author's transl)}; Katsaras K et al.; The exotoxins of Bacillus cereus show haemolytic, lethal, Phospholipase-C- and enterotoxigenic activities . The enterotoxigenic activity is regarded to be the factor which causes food poisoning in man . Efforts to purify the B . cereus exotoxins by precipitation with ammonium sulphate and subsequent chromatography on Sephadex-G-75 and Biogel-P-60 columns were partially successfull . Haemolysin and Phospholipase-C could be separated by gel-chromatography, they demonstrated partial identity on gel-diffusion agar . The lethal and enterotoxigenic activities could not be separated from the other toxins and remained heterogenous . No immunological relationship was found between Clostridium Type A alpha toxin and enterotoxin and the exotoxins of B . cereus.

J Hyg Epidemiol Microbiol Immunol, 1979, 23(3), 266 - 72
Clostridium perfringens type A: certain characters of epidemiologic significance; Chakrabarty AN et al.; Survival of spores of Cl . perfringens type A was significantly greater than that of vegetative cells in acid pH (pH 1.2) . Survival of spores in soil varied from strain to strain . Time required for 5 million spores to be reduced to 500 per gram of soil varied from 2 to 8 months with an average of 4.5+/-2.3 months . Quantitative and qualitative heat resistance studies revealed that a majority of the Indian and all the American strains tested were heat-sensitive . These characters of Cl . perfringens have an important bearing on the epidemiology of food poisoning due to this agent.

Microbiol Immunol, 1979, 23(5), 313 - 8
Effect of carbohydrates and control of culture pH on beta toxin production by Clostridium perfringens type C; Sakurai J et al.; Clostridium perfringens type C strain CN 5384 produced a higher level of beta toxin in a controlled pH medium containing 1% glucose, starch, or sucrose than in media with dextrin, fructose, or raffinose . Toxin synthesis was not related to the growth yield . The effect of glucose on beta toxin production by 11 strains was investigated with and without control of the culture pH at 7.5 . Strain CN 5386 produced distinctly higher toxin when the pH of the culture was maintained at 7.5, compared with uncontrolled pH.

Biochim Biophys Acta, 1978 Dec 20, 537(2), 185 - 207
Nitrogenase X: Mössbauer and EPR studies on reversibly oxidized MoFe protein from Azotobacter vinelandii OP . Nature of the iron centers; Zimmermann R et al.; Under anaerobic conditions the molybdenum-iron protein (MoFe protein) from Azotobacter vinelandii can be reversibly oxidized with thionine . Electron paramagnetic resonance studies reveal that the oxidation proceeds in two distinct phases: the MoFe protein can be oxidized by four electrons without loss of the EPR signal from the S = 3/2 cofactor centers . A second oxidation step, involving two electrons, leads to the disappearance of the cofactor EPR signal . In order to correlate the events during the thionine titration with redox reactions involving individual iron centers we have studied the MoFe proteins from A vinelandii and Clostridium pasteurianum with Mossbauer spectroscopy . Spectra were taken in the temperature range from 1.5 K to 200 K in applied magnetic fields of up to 54 kG . Analysis of the Mossbauer data allows us to draw three major conclusions: (1) the holoprotein contains 30 +/- 2 iron atoms . (2) Most probably, 12 iron atoms belong to two, apparently identical, iron clusters (labeled M) which we have shown previously to be structural components of the iron and molybdenum containing cofactor of nitrogenase . The M-centers can be stabilized in three distinct oxidation states, MOXe- in equilibrium MNe- in equilibrium MR . The diamagnetic (S = 0) state MOX is attained by oxidation of the native state MN with either thionine or oxygen . MR is observed under nitrogen fixing conditions . (3) The data strongly suggest that 16 iron atoms are associated with four iron centers which we propose to call P-clusters . Each P-cluster contains four spin-coupled iron atoms . In the native protein the P-clusters are in the diamagnetic state PN, yielding the Mossbauer signature which we have labeled previously 'components D and Fe2+' . Three irons of the D-type and one iron of the Fe2+-type appear to comprise a P-cluster . A one-electron oxidation yields the paramagnetic state POX . Although the state POX is characterized by half-integral electronic spin a peculiar combination of zero-field splitting parameters and spin relaxation renders this state EPR-silent . Spectroscopically, the P-clusters are novel structures; there is, however, evidence that they are closely related to familiar 4Fe-4S centers.

Biochim Biophys Acta, 1978 Dec 20, 537(2), 501 - 6
Isoelectric focusing of ferredoxins, flavordoxins and a rubredoxin; Dutton JE et al.; Isoelectric points of ferredoxins, flavodoxins and a rubredoxin from a range of sources were measured by electrofocusing over the pH range between 2.5 and 5.0 on thin layers of polyacrylamide gel . The pH gradient along the gel was measured directly by a surface electrode . The isoelectric points of the plant-type ferredoxins were between approx . 3.15 and 3.35, and those of the flavodoxins close to 3.5 . Ferredoxin, rubredoxin and flavodoxin from Clostridium pasteurianum had isolectric points of the of 2.75, 2.9, and 3.1, respectively . The values for the isoelectric points ferredoxins are significantly lower than previous results in the literature suggest.

Tijdschr Diergeneeskd, 1978 Dec 15, 103(24), 1327 - 33
{Incidence of Clostridium botulinum in the rumen contents and faeces of cattle fed brewers' grains naturally contaminated with Clostridium botulinum (author's transl)}; Notermans S et al.; The number of Clostridium botulinum type B organisms excreted by cattle fed brewers' grains in which these organisms were found to be present and the period for which they were excreted, were studied . Large numbers (10(5) - 10(7) per gramme) of these organisms were detected in the rumen contents and faeces of the animals . When feeding brewers' grains was discontinued, Cl . botulinum type B was still detectable in the faeces for a considerable period (greater than eight weeks) . There was evidence to suggest that the number of Cl . botulinum organisms multiplies in the gastrointestinal tract of cattle.

J Chromatogr Sci, 1978 Dec 10, 16(12), 623 - 9
Botulism: a pyrolysis-gas-liquid chromatographic study; Reiner E et al.; Forty samples of dried Clostridia bacteria were subjected to pyrolysis-gas-liquid chromatography (PGLC) . Examination of the key fingerprint peaks enabled the analyst to differentiate the samples into their respective antigenic groups . Peaks occurring at the high boiling end of profile could be used to distinguish proteolytic from non-proteolytic strains of C-botulinum . PGLC has proven to be a highly reproducible as well as a rapid specific method for differentiating and identifying samples of Clostridium botulinum.

Biochim Biophys Acta, 1978 Dec 8, 527(2), 359 - 69
Nitrogenase: properties of the catalytically inactive complex between the Azotobacter vinelandii MoFe protein and the Clostridium pasteurianum Fe protein; Emerich DW et al.; The catalytically inactive complex generated by the combination of the Azotobacter vinelandii MoFe protein (Av1) and the Clostridium pasteurianum Fe protein (Cp2) inhibits N2 reduction, C2H3 reduction, H+ reduction and ATP hydrolysis catalyzed by the homologous nitrogenases . Kinetic data indicate that the inactive complex consists of two molecules of Cp2 to one molecule of Av1, with values for the inhibitor constant in the range of 1--10 nM . Inhibition of C . pasteurianum nitrogenase by Av1 produces a lag phase in acetylene reduction that increases with increasing Av1 . The lag phase is found only at levels of Av1 sufficient to keep the ratio of Cp2 : Cp1 lower than 2 . Gel filtration of a mixture of Av1 and Cp2 provides evidence for complex formation and indicates that each Av1 molecule binds more than one Cp2 molecule . The Av1-Cp2 complex binds two molecules of MgATP per molecule of Cp2 . MgATP is not required for complex formation, but complex formation lowers the MgATP-Cp2 dissociation constant approx . 3-fold . Av1 protects the iron-sulfur center in Cp2 completely against the MgATP-induced reaction with chelators . This provides additional evidence for formation of the Av1-Cp2 complex and together with the results of the MgATP-binding studies suggests that the two binding sites for MgATP are some distance away from the iron-sulfur site on Cp2.

Eur J Biochem, 1978 Dec, 92(2), 449 - 54
FAD is covalently attached to peptidyl-tRNA during cell-free synthesis of 6-hydroxy-D-nicotine oxidase; Hamm HH et al.; The process, by which FAD is attached covalently to the 6-hydroxy-D-nicotine oxidase apoprotein in D-nicotine-induced cells of Arthrobacter oxidans was studied in vitro . {3H}Adenine-labelled FAD prepared biosynthetically in Clostridium kluyveri was incorporated into the 6-hydroxy-D-nicotine oxidase molecule during cell-free translation . FAD rather than FMN or riboflavin was thus shown to be the flavin derivative transferred to the polypeptide chain . After short-term protein synthesis on ribosomes from induced A . oxidans cells in the presence of an Escherichia coli MRE 600 supernatant fraction and {adenine-2-3H}FAD, THE PEPTIDYL-TRNA fraction was separated from completed polypeptides . Labelled FAD was found to be covalently attached to the tRNA-bound polypeptides . Cleavage of the tRNA-peptide bond released labelled polypeptides the largest of which migrated as authentic 6-hydroxy-D-nicotine oxidase during dodecylsulfate/polyacrylamide gel electrophoresis . These results strongly suggest that FAD is incorporated into the nascent polypeptide chains of 6-hydroxy-D-nicotine oxidase during ribosomal translation.

J Med Chem, 1978 Dec, 21(12), 1301 - 7
Synthesis and quantitative structure--activity relationships of some antibacterial 3-formylrifamycin SV N-(4-substituted phenyl)piperazinoacethydrazones; Kiritsy JA et al.; A series of 14 3-formylrifamycin SV N-(4-substituted phenyl)piperazinoacethydrazones has been synthesized and evaluated for their antimicrobial activity . The compounds were found active against Bacillus subtilis, Staphylococcus aureus, Mycobacterium phlei, and Mycobacterium tuberculosis but not as active as rifampin . The compounds also exhibited significant activity against Clostridium perfringens and in this bacterial system some were more active than rifampin . The QSAR showed that the activity against B . subtilis depended only on lipophilicity, and the regression equation was linear . A parabolic relationship between the antibacterial activity and lipophilicity of the compounds was found in Staph . aureus . Additionally, the activity was dependent upon the electronic and steric effects of the phenyl substituents . The sensitivity of M . phlei to the compounds was found to correlate well with a linear combination of hydrophobic, electronic, and steric parameters . No statistically significant correlation was possible between the physicochemical parameters studied and the activity of the compounds against C . perfringens and M . tuberculosis.

Can J Biochem, 1978 Dec, 56(12), 1141 - 8
Bile acids of a 3200-year-old Egyptian mummy; Kuksis A et al.; The bile acids of the gall bladder and hepatic tissue of a 3200-year-old Egyptian mummy were isolated by thin-layer chromatography and identified by combined gas-liquid chromatrography and mass spectrometry . Except for complete deconjugation and extensive dehydration, the bile acids were found to correspond in their qualitative and quantitative composition to the gall bladder bile acids of modern man . The secondary bile acids constituted about 50% of the total and were identified as the normal bacterial oxidoreduction products of the primary bile acids and their dehydration products . In addition a series of unsaturated bile acids were identified, which corresponded to the dehydration products of cholic and chenodeoxycholic acids . It is suggested that both bile acid deconjugation and the limited oxidoreduction were probably brought about by the Clostridium organisms identified in the tissue . On the basis of the bile acid composition it is concluded that the ancient man metabolized cholesterol along the same pathways as modern man.

Can J Microbiol, 1978 Dec, 24(12), 1602 - 6
Endotoxin in germfree, gnotobiotic, or conventional-flora Sprague-Dawley rats; McLeod JC et al.; The Limulus assay for bacterial endotoxin was performed on serum and (or) plasma from animals monoassociated with Clostridium species, Staphylococcus aureus, Escherichia coli, Proteus mirabilis, Enterobacter agglomerans, Bacteroides fragilis, Klebsiella pneumoniae, or Candida albicans . Plasma from animals monoassociated with the gram-negative bacteria or C . albicans consistently showed a positive Limulus test while conventional-flora controls, germfree rats, and gnotobiotic animals monoassociated with gram-positive bacteria or E . agglomerans were negative . Germfree and conventional rats were injected (intraperitoneal (i.p.)) with Salmonella typhosa lipopolysaccharide (LPS) . Although no endotoxin was detectable in either group prior to the injection, by 1 h post injection endotoxin was in the plasma of all groups . The germfree rats appeared to clear the LPS quicker than their conventional-flora counterparts . Generally, LPS-injected rats (conventional and germfree) showed clumping and decreased number of platelets, a decrease in their lymphocyte counts, and increased polymorphonuclear leukocyte (PMN) counts.

Eur J Biochem, 1978 Nov 15, 91(2), 345 - 50
Isolation of thiomolybdate compounds from the molybdenum-iron protein of clostridial nitrogenase; Zumft WG; Acid/base treatment of the molybdenum-iron protein of the nitrogenase from Clostridium pasteurianum 25 yields low-molecular-weight compounds of molybdenum, which can be separated from the protein by gel chromatography . Elementary analysis and spectral properties relate these compounds to thiomolybdate anions . It is proposed that in its native state nitrogenase contains a thio complex of molybdenum coupled to iron-sulfur clusters.

Lancet, 1978 Nov 11, 2(8098), 1014 - 6
Fulminant necrotising enterocolitis associated with Clostridia; Kosloske AM et al.; 5 infants with no growth of bacteria on cultures of blood and peritoneal fluid recovered from necrotising enterocolitis after medical treatment alone . 12 infants with positive cultures required surgery . 5 of these 12, who did not harbour clostridia, had a mild clinical course and all 5 survived segmental bowel resection . The 7 infants who harboured clostridia had a more severe clinical course and 4 died . In 3 of 4 infants with Clostridium perfringens, the necrotising enterocolitis was fulminant, characterised by severe pneumatosis intestinalis, extensive gangrene, early intestinal perforation, and a fatal outcome.

J Biol Chem, 1978 Nov 10, 253(21), 7722 - 30
Assignment of the cysteinyl 13C nuclear magnetic resonances and comparison of other aliphatic amino acid resonances of Clostridium acidi-urici, Clostridium pasteurianum, and Peptococcus aerogenes ferredoxins; Packer EL et al.; 13C NMR spectra of Clostridium acidi-urici, Clostridium pasteurianum, and Peptococcus aerogenes ferredoxins show that some 13C resonances of the aliphatic amino acid residues are shifted significantly from their corresponding resonance positions in the spectra of model polypeptides or apoferredoxin . Thirteen 13C resonances are shifted into the 80- to 120-ppm (from CS2) region, and have been assigned to the cysteinyl alpha and beta carbon atoms . The remaining shifted resonances in the 120- to 190-ppm region are tentatively assigned to amino acid residues that may be close to {4Fe-4S} clusters of the oxidized and reduced ferredoxins . The similarity in the shift pattern of the corresponding 13C resonances of the cysteinyl alpha and beta carbon atoms in the three ferredoxins studied suggests that the three-dimensional amino acid environments of the corresponding {4Fe-4S} clusters in each protein are similar.

Can Med Assoc J, 1978 Nov 4, 119(9), 1058 - 60
Pseudomembranous colitis: isolation of two species of cytotoxic clostridia and successful treatment with vancomycin; Marrie TJ et al.; Lincomycin-resistant Clostridium sporogenes obtained from the stools of a patient with lincomycin-associated pseudomembranous colitis produced a heat-stable cytotoxin in low titre when grown in chopped meat medium . Vancomycin eradicated this strain and all other clostridia, and controlled the symptoms . When diarrhea recurred 7 days after treatment with vancomycin was stopped, clostridia including C . sporogenes and C . difficile were again isolated . The C . difficile produced a heat-labile cytotoxin in high titre that was unaffected by growth in various media and induced colitis in hamsters . Treatment with vancomycin, to which all the clostridia were sensitive, eradicated both toxic species and controlled the diarrhea . Antibiotic-induced pseudomembranous colitis may be associated with more than one species of toxin-producing clostridia . Vancomycin therapy should be continued for 10 days or more in patients with severe disease to eradicate the responsible organism.

J Am Vet Med Assoc, 1978 Nov 1, 173(9), 1131 - 3
Hazards of disease transfer from marine mammals to land mammals: review and recent findings; Smith AW et al.; In a 5-year study (1972-1977) of microbial agents isolated from both clinically normal and diseased marine mammals, it was shown that certain disease agents are widespread in a diversity of ocean populations and that some are also transmissible to a number of terrestrial mammal species . Leptospira interrogans serovar pomona has been isolated repeatedly from 2 species of pinnipeds (Zalophus californianus califonianus and Callorhinus ursinus) . Some of the more important bacterial pathogens for land mammals that were isolated from wild marine mammals are Pseudomonas mallei, Clostridium chauvoei, C novyi, Neisseria mucosa var heidelbergensis, Klebsiella pneumoniae, Salmonella spp, and Pasteurella multocida . Numerous serotypes of viruses classified as caliciviruses were isolated from a variety of marine mammals . Some of these are known to infect several land mammal species including swine horses, and primates . For this reason., precautions should be taken to ensure that disease agents shed by captive marine mammals are not transmitted to susceptible terrestrial mammals, including animal handlers and other human beings.

J Clin Microbiol, 1978 Nov, 8(5), 509 - 11
Clostridium pseudotetanicum bacteremia in a patient with Pasteurella multocida conjunctivitis; Eschete ML et al.; Clostridium pseudotetanicum only once previously has been identified as causing disease . Pasteurella multocida has been identified only three times as the cause of purulent conjunctivitis . A very debilitated patient had C . pseudotetanicum bacteremia and P . multocida conjunctivitis from which she recovered only to die of a nosocomial Staphylococcus epidermidis septicemia, originating in a site for the administration of intravenous fluids.

Infect Immun, 1978 Nov, 22(2), 418 - 22
Neutralization of Clostridium difficile toxin by Clostridium sordellii antitoxins; Chang TW et al.; Neutralization of Clostridium difficile toxin by Clostridium sordellii antitoxin was studied by cytotoxicity assay in tissue culture . The sources of toxin were stools from two patients with pseudomembranous colitis and a culture filtrate of C . difficile isolated from one of the patients . C . sordellii antitoxin was available either in monovalent form or as gas gangrene polyvalent antitoxin . The potency of antitoxins against C . difficile determined by cytotoxicity assay did not correlate with the established values reported for mouse protection tests against C . sordellii toxin . An equivalent zone of optimal neutralization was demonstrated for stool toxin, and a slightly different one for culture toxin . The rate of neutralization appeared to be instantaneous, either at 24 or at 37 degrees C . The efficacy of antitoxin in preventing cytotoxicity in cultured cells preexposed to toxin decreased rapidly with preexposure time . The union between toxin and antitoxin could be readily dissociated by simple dilution or by ammonium sulfate precipitation followed by dissociated by simple dilution or by ammonium sulfate precipitation followed by dilution . Continued incubation of toxin-antitoxin mixture did not increase the firmness of the union; on the contrary, more dissociation occurred . The unusual looseness of the toxin-antitoxin union is probably relatd to lack of serological specificity or affinity . Based on these observations, a practical diagnostic method for antibiotic-induced colitis is outlined.

Zh Mikrobiol Epidemiol Immunobiol, 1978 Nov, (11), 59 - 62
{Mechanism of segregation of bacterial chromosomes following division of anaerobic clostridia}; Vaisman ISh; Morphological events during division in Clostridium oedematiens, strains A-277 and A-79, were studied on ultrathin sections . The bacterial cells of the species under investigation containing numerous and various intracytoplasmic membranous structures were practically devoid of the "nuclear mesosomes" type structures . In these anaerobic Clostridia the segregation of bacterial chromosomes after the replication was effected by means of direct connections between the DNA filaments and the cytoplasmic membrane of the bacterial cell . These connections were multiple, up to 250 nm in length, possibly ensuring a proper mechanical strength needed for translocation of the chromosomes into the daughter cells . It is suggested that multiple contact zones between the cytoplasmic membrane and the DNA filaments of the bacterial chromosome arose after the triggering of their DNA replication.

Arch Intern Med, 1978 Nov, 138(11), 1618 - 20
Ticarcillin disodium in anaerobic infections; Webb D et al.; Twenty-five patients were treated with ticarcillin disodium, 18 of whom had anaerobic infections that included pleuropulmonary infections (seven), mandibular osteomyelitis (four), perirectal abscess (two), sepsis, primary site unknown (one), liver abscess (one), pelvic abscess (one), decubitus ulcer (one), and synergistic gangrene (one) . Seven had no anaerobic infections . Three had anaerobic septicemia . Culture results included anaerobes: peptococci (ten), peptostreptococci (ten), Bacteroides fragilis (six), Bacteroides not fragilis (ten), eubacteria (three), fusobacteria (two), Clostridium (one), Veillonella (one), and acidaminococcus (one); aerobes: Proteus (three), Klebsiella (two), Escherichia coli (two), and streptococci (two) . Six patients with mixed aerobic infections initially received gentamicin sulfate in addition . The serum levels were 110 +/- 20 microgram/ml one hour after intravenous infusion of 5 g of ticarcillin disodium . All anaerobic isolates were susceptible at less than or equal to 100 microgram/ml and 85% by less than or equal to 25 microgram/ml of ticarcillin . Sixteen patients responded well to ticarcillin and two failed to respond . Our study suggests that ticarcillin is useful in the treatment of anaerobic infections.

Gastroenterology, 1978 Nov, 75(5), 778 - 82
Role of Clostridium difficile in antibiotic-associated pseudomembranous colitis; Bartlett JG et al.; Tissue cultures were performed on stools from 189 patients to detect a cytopathic toxin which is neutralized by Clostridium sordellii antitoxin . Specimens satisfying these criteria were considered positive in the tissue culture assay . Stools from 26 of 27 patients with antibiotic-associated pseudomembranous colitis were positive and 16 of these specimens showed toxin titers of 10(-3) dilutions or greater . The tissue culture assay was positive with specimens from 9 of 63 patients with antibiotic-associated diarrhea without documented pseudomembrane formation . Stools from patients with neonatal necrotizing enterocolitis, ulcerative colitis, and healthy controls were uniformly negative in this assay . Cultures were performed on stools from 38 patients with antibiotic-associated diarrhea or colitis to detect clostridia which produce a cytopathic toxin in vitro . Clostridium difficile was recovered from 6 of 8 specimens which were positive in the tissue culture assay and 5 of 30 which were negative in this assay . C . sordellii was recovered in a single specimen . One hundred and nine clostridia strains were tested in the tissue culture assay and C . difficile was the only species which produced a cytopathic toxin . All strains of this organism were positive in the tissue culture assay and, in each instance, cytotoxicity was neutralized by C . sordellii antitoxin . These results indicate that C . difficile is the major cause of antibiotic-associated pseudomembranous colitis and offer an explanation for previous studies showing that the cytotoxin of stools from these patients is neutralized by C, sordellii antitoxin.

Gastroenterology, 1978 Nov, 75(5), 791 - 5
Pathogenesis of mucosal injury in the blind loop syndrome; Jonas A et al.; Bacterial extracts were prepared from cultures originating in chronic self-filling intestinal blind loops in rats . Their ability to remove active maltase molecules from isolated brush border membranes was studied in vitro . Twelve strains in 51 tested, belonging to one of three species, Bacteroides fragilis, Clostridium perfringens, and Streptococcus fecalis, possessed maltase-releasing activity . The ability to remove maltase correlated well with the ability to hydrolyze p-nitrophenyl-tert-butyloxycarbonyl-l-alaninate (NBA), an ester substrate rapidly hydrolyzed by elastase, but not with substrated favored by tryhsin and chymotrypsin . Maltase-releasing activity from C . perfringens was strongly inhibited by soybean trypsin inhibitor and to a lesser extent by lima bean trypsin inhibitor . Of four chloromethylketone active-site directed inhibitors tested with specificities for elastase, trypsin, and chymotrypsin, inhibition was maximal with elastase-specific inhibitors . In two species, activity was shown to be heat sensitive, and to be inhibited by concentration of the extract . In one species maltase-releasing activity was shown to be due to an enzyme of molecular weight at least 66,000 with the capacity to remove lactase, sucrase, and alkaline phosphatase, as well as maltase . The results indicate that anaerobic or facultatively anaerobic species, previously identified with the pathology of of the blind loop syndrome, contain proteases which are capable of removing components of the intestinal surface membrane . These proteases appear to have elastase-like substrate specificity and may be involved in the etiology of disaccharidase deficiency in bacterial overgrowth syndromes.

Proc Natl Acad Sci U S A, 1978 Nov, 75(11), 5273 - 5
Cluster characterization in iron-sulfur proteins by magnetic circular dichroism; Stephens PJ et al.; We report magnetic circular dichroism (MCD) spectra of 4-Fe iron-sulfur clusters in the iron-sulfur proteins Chromatium high-potential iron protein (HIPIP), Bacillus stearothermophilus ferredoxin and Clostridium pasteurianum ferredoxin . The MCD is found to vary significantly with cluster oxidation state but is relatively insensitive to the nature of the protein . The spectra obtained are compared with the corresponding spectra of iron-sulfur proteins containing 2-Fe clusters . It is concluded that MCD is useful for the characterization of iron-sulfur cluster type and oxidation state in iron-sulfur proteins and is superior for this purpose to absorption and natural circular dichroism spectroscopy.

Aust Vet J, 1978 Nov, 54(11), 541 - 4
Reverse phase passive haemagglutination and single radial immunodiffusion to detect epsilon antigen of Clostridium perfringens type D; Beh KJ et al.; Two in vitro immunological assays were developed for detection of the epsilon (epsilon) antigen of Cl . perfringens type D . It was found that the reverse phase passive haemagglutination assay (RPHA) was able to detect concentrations of epsilon-antigen as low as 6 x 10-7 mg/ml whereas the single radial immunodiffusion techniques (SRID) was capable of detecting concentrations of epsilon-antigen above 0.01 mg/ml . When applied to gut contents from freshly dead infected sheep the RPHA test was found to be more sensitive than mouse toxicity assay in detecting the presence of epsilon-antigen . However, very low titres were detected in gut contents from normal sheep which meant that in a diagnostic situation interpretation of RPHA titres would be difficult . No epsilon-antigen was detected by SRID in gut contents from normal sheep or in gut contents from freshly dead infected sheep . The SRID assay could detect epsilon-antigen in gut contents from infected sheep allowed to decompose for 20 h post-mortem.

Endokrinologie, 1978 Nov, 72(3), 363 - 4
Specificity of action of neuraminidase, according to its bacteriological origin; Finne E; Neuraminidase from Vibrio Cholerae selectively cleaves sialic acid from FSH, but leaves the LH sialic acid, not influencing the biological activity of the latter hormone . On the other hand, Neuraminidase from Clostridium perfringens does not possess this specific action and destroys the biological activity of LH as is suggested by Parlow's OAAD-test.

J Clin Microbiol, 1978 Nov, 8(5), 480 - 8
Synergistic hemolysis phenomenon shown by an alpha-toxin-producing Clostridium perfingens and streptococcal CAMP factor in presumptive streptococcal grouping; Gubash SM; A new phenomenon of synergistic hemolysis by Clostridium perfringens alpha-toxin and the streptococcal CAMP factor on human and guinea pig erythrocytes is described . A possible mode of action of the CAMP factors is suggested . On human blood agar all of the tested isolates of group B streptococci gave an arrowhead-shaped zone of hemolysis; 74% of group A gave a crescent-shaped lytic zone, whereas all isolates of groups C and G and the remaining 26% of group A streptococci gave a bullet-shaped lytic zone . By comparison, in the CAMP test incubated aerobically and anaerobically, 70 and 91%, respectively, of streptococci other than group B gave positive, arrowhead-shaped lytic zones . If all intermediate positive reactions in the CAMP tests were read as negative after aerobic incubation, only 89% of group B streptococci would be properly identified . The synergistic hemolysis phenomenon, using an alpha-toxin-producing C . perfringens and human blood agar, provided a reliable test for presumptive identification of group B streptococci, with promising potential to differentiate in the same test group A streptococci from other groups.

Mikrobiologiia, 1978 Nov-Dec, 47(6), 1124 - 6
{Number of butyric acid bacteria belonging to the genus Clostridium in the slimy sediments of Volga reservoirs}; Dziuban AN; The number of Clostridium pasteurianum, Cl . butyricum, and Cl . acetobutylicum was determined in ooze deposits of the Volga River reservoirs using enriched nutrient media . The bacterial number for the two former species was about 1 + 10(6) cells per 1 ml of ooze with a high content of easily assimilated organic substances, thus being by 1--3 orders of magnitude higher than for Cl . pasteurianum on media without nitrogen . The bacterial number for Cl . acetobutylicum was 0.1--1 + 10(3) cells per 1 ml.

Biochemistry, 1978 Oct 31, 17(22), 4770 - 8
Circular dichroism and magnetic circular dichroism of iron-sulfur proteins; Stephens PJ et al.; Circular dichroism (CD) and magnetic circular dichroism (MCD) spectra are reported for the 2-Fe ferredoxins from Pseudomonas putida and Spirulina maxima, Chromatium HIPIP, the 4-Fe ferredoxin from Bacillus stearothermophilus, and the 8-Fe ferredoxin from Clostridium pasteurianum . The spectral range spans the near-infrared, visible, and near ultraviolet . In all cases except oxidized 2-Fe ferredoxins, electronic absorption is observed continuously from less than 5000 cm-1 to above 30,000 cm-1 . The CD spectra of the two 2-Fe ferredoxins are similar . In contrast, the CD of the 4-Fe and 8-Fe proteins, for a given 4-Fe cluster oxidation level, varies considerable with protein . MCD is less sensitive to protein environment than is CD . In the 2-Fe proteins, MCD at 5 T is appreciably smaller than the CD; in the 4-Fe and 8-Fe proteins, MCD and CD are comparable in magnitude . Both CD and MCD are more highly structured than the corresponding absorption spectra . The CD and MCD spectra reported provide a broader base than heretofore available for the characterization of iron-sulfur proteins containing 2-Fe and 4-Fe clusters and for the evaluation of electronic structural models for these clusters.

Wien Klin Wochenschr, 1978 Oct 27, 90(20), 733 - 6
{Detection of collagenase in passive haemagglutination using collagen-coated erythrocytes (author's transl)}; Kojer M et al.; Human rheumatoid arthritis (RA) collagenase and bacterial collagenase were shown to agglutinate collagen-coated erythrocytes . Native collagens of type I and type II reacted equally well, while denatured collagens showed less distinct agglutination activity . The sensitivity of the method for the detection of purified bacterial collagenase from Clostridium histolyticum is very high (100 pg) . It is, however, low for human RA collagenase . The agglutination reaction is not inhibited by concentrations of native collagen causing distinct inhibition of anticollagen sera (2mg%) . EDTA inhibits the agglutination completely.

Arch Microbiol, 1978 Oct 4, 119(1), 7 - 11
Fumarate reductase of Clostridium formicoaceticum . A peripheral membrane protein; Dorn M et al.; When Clostridium formicoaceticum was grown on fumarate or L-malate crude cell extracts contained a high fumarate reductase activity . Using reduced methyl viologen as electron donor the specific activity amounted to 2-3.5 U per mg of protein . Reduced benzyl viologen, FMNH2 and NADH could also serve as electron donors but the specific activities were much lower . The NADH-dependent activity was strictly membrane-bound and rather labile . Its specific activity did not exceed 0.08 U per mg of particle protein . Fumarate reductase activity was also found in cells of C . formicoaceticum grown on fructose, gluconate, glutamate and some other substrates . The methyl viologen-dependent fumarate reductase activity could almost completely be measured with intact cells whereas only about 25% of the cytoplasmic acetate kinase activity was detected with cell suspensions . The preparation of spheroplasts from cells of C . formicoaceticum in 20 mM HEPES-KOH buffer containing 0.6 M sucrose and 1 mM dithioerythritol resulted in the specific release of 88% of the fumarate reductase activity into the spheroplast medium . Only small amounts of the cytoplasmic proteins malic enzyme and acetate kinase were released during this procedure . There results indicate a peripheral location of the fumarate reductase of C . formicoaceticum on the membrane.

Biochim Biophys Acta, 1978 Oct 3, 543(2), 226 - 34
Studies on the characterization of the Rho(D) antigen; Litten J et al.; The Rho(D) antigen of red cell membranes was solubilized using ethylene-diamine tetraacetic acid (EDTA) and 2-mercaptoethanol . The solubilized antigen was partially separated from other solubilized membrane components using molecular filtration . The antigen was treated with various enzymes to learn some of the chemical characteristics . It was found that the activity of the antigen, as measured by hemagglutination inhibition, was not affected by bee venom phospholipase A, Clostridium welchii phospholipase C, calf-intestinal alkaline phosphatase, Vibrio cholerae neuraminidase, pig kidney leucine aminopeptidase, bovine pancreatic carboxypeptidase A, and pig pancreatic carboxypeptidase B . However, the proteolytic enzymes, pronase, trypsin, chymotrypsin and papain, did destroy Rho(D) activity as measured by hemagglutination inhibition . These results indicate that protein is an important part of the active determinant of the Rho(D) antigen . The experiments by other investigators have shown that lipid is important to maintain the Rho(D) activity in the intact membrane; lipid probably helps to maintain the structural conformation of the Rho(D) molecule in its natural environment . The solubilized Rho(D) molecules are apparently not dependent on lipid for their Rho(D) activity.

Am J Clin Nutr, 1978 Oct, 31(10 Suppl), S243 - S247
Characterization of 7-alpha-dehydroxylase in Clostridium leptum; Stellwag EJ et al.; 7-alpha-Dehydroxylation of primary bile acids was demonstrated radiochromatographically in whole cells of Clostridium leptum but was not observed in intestinal Bacteroides species . Activity of 7-alpha-Dehydroxylase was detected within a pH range of 5 to 9 and was 8-fold higher in specific activity in cell cultures in the presence of 0.1 mM sodium cholate than in its absence . 7-alpha-Dehydroxylase activity in whole cells was markedly inhibitied by 2,4-dinitrophenol, carbonyl-cyanide-m-chlorophenylhydrazine, and dicyclohexylcarbodiimide . A hypothesis concerning the dietary regulation of 7-alpha-dehydroxylating intestinal anaerobic bacteria is presented.

Ann Microbiol (Paris), 1978 Oct, 129 B(3), 437 - 49
{Investigation of the immunity status towards tetanus of a population of mechanics at the car-factory "Renault" (author's transl)}; Bizzini B et al.; Tetanus immunity has been studied in a population of mechanics working at the car-factory "Renault" . For the study, 283 individuals were divided into 3 groups . The first group consisted of non-vaccined individuals, the second of vaccinated individuals and the third of individuals who had been boostered . The influence of different parameters on tetanus immunity status was considered, i . e . the age of the subjects, the time at which boostering was given, the serotherapy (when it was administered) and the contact with machine lubricating and cooling fluids . Clostridium tetani bacilli and spores were frequently found in aqueous machine fluids . Their presence is a potential hazard for non-vaccinated mechanics coming to contact with the fluids . Tetanus antibody levels in the sera of the tested population were determined in vivo by the toxin neutralization test . The influence on tetanus immunity of the different parameters considered in this paper was subjected to statistical analysis . From the whole population, 27% of the individuals were found to have no protection against tetanus . When age was taken into account, 53% of the individuals over 45 years old and 10% of those under 45 years old were shown to be devoid of tetanus immunity . It could be shown that younger individuals were better protected than older ones, because the formers had been immunized with adsorbed tetanus toxoid and most of the latters with fluid toxoid . Of the individuals in the third group who had received a booster injection within 15 years after primo-vaccination, 98% showed protective tetanus antibody levels in their sera in contrast to 25% when boostering had occurred more than 15 years after primo-vaccination . Contact with machine fluids was found to influence the degree of immunity of only those individuals whose boostering dated back to more than 25 years . Unexpectedly, 3 mechanics seemed to develop immunity after coming into contact with machine fluids . From the results reported here, it is concluded that tetanus immunity in vaccinated individuals should be renewed by a compulsory booster injection given every 5, 10 or, at the minimum, 15 years . Moreover, high-risk populations such as that represented by the mechanics should be immunized or boostered on commencing employment.

Zentralbl Bakteriol {Orig A}, 1978 Oct, 241(4), 438 - 47
{Characterization of plasmid DNA in a lecithinase-positive and in a lecithinase-negative strain of Clostridium perfringens (author's transl)}; Kramer J et al.; A non pathogenic variant of Clostridium perfringens and the wild type strain were characterized . The strains agreed in most of the biochemical properties, in susceptibility against antibiotics and in bacteriocin production . Contrary to the wild type the variant did not produce lecithinase and gelatinase . In deoxyribonucleic acid (DNA) of both strains centrifuged in cesiumchlorid-ethidiumbromide equilibrium there was found a satellite peak containing three distinct, covalently closed circular (CCC) DNA elements . The sum of the average molecular weight or contour length of the two small circular molecules was equal to the average molecular weight or contour length of the third . The presence of the plasmids in the variant indicated that the synthesis of lecithinase might not be coded by a plasmid.

Lab Anim Sci, 1978 Oct, 28(5), 536 - 40
Enterotoxemia in rabbits; Patton NM et al.; The presence of Clostridium perfringens Type E iota toxin was confirmed from the cecal contents of 23 of 46 rabbits which died of enteritis complex . The most consistent lesions observed were hemorrhage and edema in the cecum . Rabbit toxicity tests showed the toxic cecal contents were lethal for young rabbits unless incubated with Clostridium perfringens Type E antiserum.

Nord Vet Med, 1978 Oct, 30(10), 430 - 3
{A simple device for cultivation of anaerobic bacteria (author's transl)}; Melinder PC; A new concept for isolation and enumeration of anaerobic bacteria in food is presented . The sample is collected and diluted under anaerobic conditions in a specially designed syringe (M.O.S.), in which cultivation also takes place . A comparison between this method and cultivation in anaerobic jars (GasPak) from a common inoculum revealed a higher number of Clostridium perfringens with the M.O.S.-technique.

Appl Environ Microbiol, 1978 Oct, 36(4), 567 - 71
New medium for rapid screening and enumeration of Clostridium perfringens in foods; Erickson JE et al.; A rapid and sensitive procedure for estimating low numbers of Clostridium perfringens has been investigated and compared to methods used currently in the food industry . The new liquid medium, RPM (rapid perfringens medium), was compared with sulfite-polymyxin-sulfadiazine agar and tryptose-sulfite-cycloserine agar in recovery studies with naturally contaminated and with inoculated foods . The medium consists of a mixture of litmus milk and fluid thioglycolate medium fortified with glucose, peptone, gelatin, yeast extract, sodium chloride, and ferrous sulfate . Selectivity is based on an antibiotic system (polymyxin B sulfate and neomycin sulfate) incorporated into the medium, coupled with an incubation temprature of 46 to 48 degrees C for 24 h . Tubes were scored as positive if a stormy fermentation was observed . All tubes demonstrating stormy fermentation were confirmed as containing C . perfringens . Of a total of 774 naturally contaminated food samples, 546 samples (71%) were found to contain C . perfringens with RPM, whereas only 168 (22%) of the samples were positive using sulfite-polymyxin-sulfadiazine agar . C . perfringens was isolated from 71% of 85 other samples using RPM as compared to 14% with tryptose-sulfite-cycloserine agar . Enumeration studies on 14 individual samples using the most probable number technique also demonstrated greater sensitivity with RPM.

Fed Proc, 1978 Oct, 37(12), 2577 - 81
Foodborne hazards of microbial origin; Foster EM; Foods can serve as vehicles of many pathogenic and toxigenic agents of disease . Bacterial agents comprise three groups: 1) those that grow in the food and produce an active toxin before consumption (e.g., clostridium botulinium); 2) those that merely exist as contaminants in the food but are able to initiate infection when swallowed (e.g., Salmonella spp.); and 3) those that multiply and produce large numbers of vegetative cells in the food, then release an active enterotoxin when they sporulate in the gut . A few parasitic (e.g., Trichinella spiralis) and viral agents (e.g., hepatitis A) also can be transmitted by food . Botulinum poisoning is the deadliest foodborne disease . The potential danger of botulism from cured meats is a major factor in the argument over use of nitrites as meat curing agents . A new disease called infant botulism has been recognized since 1976 . Apparently it is not foodborne but results from intraintestinal growth of C . botulinum in very young infants . Salmonellosis is the most important of the foodborne diseases from the standpoint of overall human health . The primary vehicles are contaminated raw meat, poultry, and eggs . Faulty food handling practices are responsible for most food poisoning in the United States.

Biochimie, 1978 Sep 29, 60(6-7), 653 - 61
Phosphatidic and lysophosphatidic acid production in phospholipase C-and thrombin-treated platelets . Possible involvement of a platelet lipase; Mauco G et al.; Incubation of 32P-labelled platelets with Clostridium welchii phospholipase C greatly stimulates 32P-incorporation into phosphatidic and lysophosphatidic acids . A net synthesis is demonstrated for both phospholipids, which exhibit identical specific radioactivities . Phosphatidic acid production roughly parallels the phospholipase C-induced aggregation, whereas lysophosphatidic acid appears secondarily during cell lysis . The same qualitative variations are observed during thrombin-induced aggregation . At the physiological pH used throughout the incubations, platelets display no phospholipase A activity towards phosphatidic acid, whereas diglycerides are deacylated by platelet lysates . On the basis of these findings, a mechanism for phosphatidic and lysophosphatidic acid production is proposed, involving a phosphorylation of the di- and monoglycerides formed upon phospholipase C and lipase action . The possible role of such a pathway in regulating arachidonic acid release from phospholipids during platelet activation is discussed.

C R Acad Sci Hebd Seances Acad Sci D, 1978 Sep 25, 287(6), 659 - 61
{Initiation of germination of Clostridium difficile spores by lysozyme}; Ionesco H; The germination rate of spores of C . difficile which is usually lower than 10(-5) is raised to about 5.10(-3) in presence of lysozyme . All spores are initiated by lysozyme when previously treated by sodium thioglycolate . These spores are indeed lysozyme-dependent for germination.

Biochim Biophys Acta, 1978 Sep 11, 526(1), 34 - 41
Hydrogen bonding of flavoprotein . I . Effect of hydrogen bonding on electronic spectra of flavoprotein; Nishimoto K et al.; The effect of hydrogen bonding on the transition energy and the oscillator strength of the isoalloxazine nucleus of flavins was studied by the molecular orbital method . Among the possible hydrogen bondings examined, characteristic spectral shifts were found for the hydrogen bondings at N(1) and N(5) of the nucleus . The hydrogen bonding at N(1) resulted in the shift of the first absorption band towards blue and that of the second one towards red . On the other hand, the hydrogen bonding at N(5) resulted in the shifts of both the first and the second band towards red . The spectral characteristics reported on Clostridium MP and Desulfovibrio vulgaris flavodoxin coincided with the calculated results . The application of the calculated results to D-amino acid oxidase (D-amino acid: oxygen oxidoreductase (deaminating), EC 1.4.3.3) led to the conclusion that hydrogen bonding occurs at O(12), N(3)H, O(14) and N(5) of the isoalloxazine nucleus . The occurrence of hydrogen bondings at O(12), N(3)H, and O(14) is favorable for N(5) of the isoalloxazine nucleus to accept electron from an electron donor.

Infect Immun, 1978 Sep, 21(3), 989 - 93
Cytotoxic activity of Aeromonas hydrophila; Donta ST et al.; Most strains of Aeromonas hydrophila tested demonstrated cytotoxic activity on several tissue-cultured cell lines . The cytotoxin is heat-labile, non-dialyzable, and immunologically distinct from that of Shigella dysenteriae and Clostridium perfringens . None of the aeromonas isolates was found to be enterotoxigenic by either tissue culture or rabbit ileal loop assays.

Am J Vet Res, 1978 Sep, 39(9), 1525 - 30
Antibiotic-induced lethal enterocolitis in hamsters: studies with eleven agents and evidence to support the pathogenic role of toxin-producing Clostridia; Bartlett JG et al.; Clindamycin-induced enterocolitis in hamsters was studied, using a tissue culture assay to detect clostridial toxin . It was found that animals with lethal enterocolitis had a cytopathogenic substance in cecal contents and blood that was neutralized by clostridial antitoxins . Cultures of the cecal flora yielded numerous species of clostridia, but only 1 organism was detected which produced a toxin which was cytopathic in tissue culture . This organism, Clostridium difficile, was consistently present in high concentrations, and the cell-free supernate of these strains caused enterocolitis if injected intracecally into hamsters . Ten additional antimicrobials were tested ih hamsters . Ampicillin, vancomycin, erythromycin, cephalosporins, and oral gentamicin caused lethal enterocolitis in most recipients, and all animals which died had evidence of clostridia toxin in cecal contents at necropsy . Tetracycline and metronidazole were well tolerated, and the animals given these antimicrobials had no evidence of the toxin . We conclude that toxin-producing clostridia are responsible for lethal enterocolitis due to a variety of antimicrobials in hamsters.

Appl Environ Microbiol, 1978 Sep, 36(3), 403 - 7
Sensitization of Clostridium perfringens spores to heat by gamma radiation; Gombas DE et al.; Spores of Clostridium perfringens, type A, were given separate or sequential treatments of gamma radiation (0 to 0.7 Mrad) and/or high temperature (93 to 103 degrees C) . Prior heating, sufficient to inactivate 40 to 99% of the viable spores, had no effect on the subsequent radiation inactivation rate . Prior irradiation had a sensitizing effect on subsequently heated spores . The degree of sensitization to heat, as measured by thermal inactivation rate, increased with increased radiation pretreatment dose.

Lab Invest, 1978 Sep, 39(3), 210 - 8
The effects of Clostridium perfringens enterotoxin on rat and rabbit ileum: an electron microscopic study; McDonel JL et al.; Intestinal epithelial damage caused by Clostridium perfringens enterotoxin in rats and rabbits was identified by light microscopy and compared at the surface (scanning electron microscopy), and the ultrastructural (transmission electron microscopy) levels . Under the light microscope damage to the epithelial layer of villus tips was clearly evident in cross-sections . Whole tissue viewed under the scanning electron microscope showed comparable tip localization of morpholigic damage in the form of collapsed tips and a dense covering of rounded blebs on the tips . Ulstructuctural observations included partial and sometimes complete disappearance of microvilli structures, budding of the terminal web region into the lumen, and even complete destruction of epithelial cells . These data suggest that C . perfringens enterotoxin attacks the epithelial cells with a preference for cells at the villus tips and causes damage at least in part by altering the cells' apical membranes . This then leads to cellular sloughing, death, and lysing.

Biochem J, 1978 Sep 1, 173(3), 831 - 9
Electron-paramagnetic-resonance studies on the redox properties of the molybdenum-iron protein of nitrogenase between +50 and -450 mV; O'Donnell MJ et al.; The midpoint potentials, Em, for the oxidation of the characteristic e.p.r . signal with g values near 4.3, 3.7 and 2.01, of the nitrogenase Mo-Fe proteins from a number of bacteria were measured . They were 0mV for Clostridium pasteurianum, -42mV for Azotobacter chroococcum and Azotobacter vinelandii, -95mV for Bacillus polymyxa and -180mV for Klebsiella pneumoniae Mo-Fe proteins at pH 7.9 . The oxidations were thermodynamically reversible for the proteins from A . chroococcum, A . vinelandii and K . pneumoniae and the Em was independent of protein activity for this last protein . The protein from C . pasteurianum required a lower potential for reduction than for oxidation, and the oxidation of the protein from B . polymyxa was only 70% reversible . The apparent Em of the latter protein was decreased by 40mV in the presence of 60mM-MgCl2 . The pH-dependence of the Em of the protein from K . pneumoniae was interpreted in terms of a single ionization, not directly associated with the e.p.r.-active centre, with a pKa of 7.0 in the oxidized form of the protein and a pH-independent region at low pH (Em = 118 +/- 6.3 mV) . Approx . 20% increase in activity after oxidation was observed for the proteins from B . polymyxa, A . chroococcum and K . pneumoniae . The significance of the above results and their relationship to other published data are discussed.

J Biol Chem, 1978 Aug 25, 253(16), 5832 - 8
Purification of the "corrinoid" enzyme involved in the synthesis of acetate by Clostridium thermoaceticum; Welty FK et al.; A corrinoid enzyme has been purified to approximately 80% homogeneity from Clostridium thermoaceticum . It catalyzes the formation of acetate from N5-methyltetrahydrofolate and pyruvate in combination with the required supplementary enzymes which are supplied by an extract that has been treated with propyl iodide . The enzyme was purified by chromatography on a folate affinity column and a DEAE-Bio-Gel column and by ultrafiltration . The molecular weight as determined by sedimentation equilibrium is 158,000 and the sedimentation coefficient is 10.5 S . By gel electrophoresis in sodium dodecyl sulfate, the subunit molecular weight was found to be 40,000, thus, the enzyme may be a tetramer of four similar subunits . The results of electron microscopy confirmed the tetrameric structure . In the absence of sodium dodecyl sulfate, two bands of similar intensity were observed by electrophoresis, but both yielded the 40,000 molecular weight subunit in the presence of sodium dodecyl sulfate . These results indicate the two bands represent either two different molecular weight forms of the enzyme or two differently charged isoenzymes . The enzyme is quite labile being sensitive to dilution, aerobic conditions, and light . Dithiothreitol and glycerol were found to stabilize the enzyme . The cofactor requirements for acetate synthesis have been determined . ATP, thiamin pyrophosphate, S-adenosylmethionine, and Fe2+ were found to be required for maximum activity and the Km values were determined . High concentrations of methyltetrahydrofolate, pyruvate, and S-adenosylmethionine were found to inhibit the synthesis of acetate.

Fortschr Med, 1978 Aug 10, 96(30), 1510 - 7
{Microbiological evaluation of cefoxitin, a new beta-lactamase resistant cephamycin antibiotic, compared to cephalothin, cephaloridine and cefazolin}; Schassan HH et al.; Cefoxitin (Mefoxitin), Cephaloridin, Cephalothin and Cefazolin were tested in vitro against 380 clinical isolates of Staph . aureus and Enterobacteriaceae using the tube dilution procedure . Of the gramnegative microorganisms a selection of 150 strains was examined regarding susceptibility to the antibiotic combinations cefoxitin/gentamicin and cefoxitin/sisomicin in comparison to cephalothin/gentamicin and cephalothin/sisomicin . Cefoxitin has a broad antimicrobial spectrum and a high beta-lactamase-resistance . Cefoxitin was more active than the other antibiotics tested against E . coli, Klebsiella, Serratia, Enterobacter, indole-negative and -positive Proteus species . The high effectiveness of cefoxitin against anaerobic bacteria such as Bacteroides fragilis and Clostridium perfringens is discussed . Cefoxitin has an excellent bactericidal potency . Among the combinations tested cefoxitin/sisomicin was most active and obtained synergistic effects in most species . The resistance rate of cefoxitin of 8.7% was reduced to 0.7% with cefoxitin/sisomicin.

Vet Rec, 1978 Aug 5, 103(6), 116 - 7
Haemorrhagic gastroenteritis in the dog associated with Clostridium welchii; Prescott JF et al.; Two cases of peracute haemorrhagic enteritis in the dog are reported . Gram-positive bacilli, which were shown in one case to be Clostridium welchii were found adhering to the necrotic epithelial surfaces in parts of the gastrointestinal tract in both cases . Large numbers of C welchii were recovered from the intestines of both dogs.

J Mol Evol, 1978 Aug 2, 11(3), 233 - 43
The evolution of protein sequences by repetitious gene duplication: clostridial flavodoxin; Kobayashi K et al.; Internal regularities of amino acid sequences of flavodoxins, FMN-containing, low molecular weight flavoproteins, were statistically examined using the minimum mutation method . The sequence of Clostridium pasteurianum flavodoxin shows statistically significant evidence of repetitious internal gene duplications at different levels of structure . Peptide pairs with a low chance probabilitiy of occurrence were frequently observed at a shift of 5 residues . The pairs with the lowest chance probabilities are a pair of heptapeptides at positions39--45 vs . 44--50, a 5 residue shift (p = 9 x 10(-6)) . Most of the related pairs are consistent and could best be explained by the repeating pentapeptide sequence: (Lys-Gly-Ala-Asp-Val-)n and appropriate gaps . Internal repetitions with longer shifts were also suggested for other flavodoxins . Repetitious gene duplication is proposed for the early stages of flavodoxin evolution.

Zentralbl Bakteriol {B}, 1978 Aug, 167(1-2), 22 - 8
{Investigation of the hygienic standard in two hospitals including the control of disinfection (author's transl)}; Pfanzelt R et al.; In two operative departments with different architectural presuppositions, the hygienic standard was checked up . Under favourable conditions in clinic B (Hosch-filter, sluice-systems) the relative frequency of demonstrable bacteria amounted to 55% . In clinic A, where these conditions failed, it amounted to 80% . Among the non pathogenic bacteria DNase-negative staphylococci were demonstrated more frequently than others . 13.4% and 18.9% resp . of the bacteria were DNase-positive staphylococci . We used Clostridium perfringens for detecting invasion-paths of germs . The most important ones are leaky windows, air conditioning and insufficient sluice-systems . The success of desinfection was examined . It fluctuates from 67% to 100% . One control amounted to 42% . The results show, that it is impossible to establish sterile rooms for common operative departments . But they show as well that a satisfying hygienic standard cannot be arrived without sluice-systems and appropriate air conditioning.

J Lipid Res, 1978 Aug, 19(6), 757 - 62
Effects of small amounts of pentadecan-2-one on the growth of Clostridium butyricum; Gilbertson JR et al.; Primary alcohols occur as trace lipids and are the only long-chain alcohol species present in Clostridium butyricum . Secondary alcohols do not occur physiologically in this microorganism . Exposure of these cells to the methyl ketone, pentadecan-2-one, results in a marked decrease in the primary alcohol content with the secondary alcohol, pentadadecan-2-ol, becoming the major alcohol present . This change in lipid composition is associated with a significant decrease in growth rate that is proportional to the log of the pentadecan 2-one concentration of the incubation medium . When these cells are incubated with pentadecan-2-ol alone, growth is unaffected . Simultaneous exposure of the bacteria to pentadecan-2-one and a mixture of primary alcohols results in a partial relief of the growth inhibition observed with the ketone alone . These observations indicate that pentadecan-2-one inhibits the formation of primary alcohols that are important for normal growth of this bacterium.

J Antibiot (Tokyo), 1978 Aug, 31(8), 756 - 60
In vitro activity of tiamulin (81.723 HFU), a new pleuromulin derivative, against clinically significant anaerobes; Werner H et al.; The susceptibility of more than 40 strains of Gram-negative and Gram-positive anaerobes to tiamulin (Sandoz 81.723 hfu), a new pleuromulin (pleuromutilin) derivative, was determined by broth dilution and agar dilution tests . The influences of density of the inoculum upon MICs was studied by a specially designed pour plate-technique . Bacteroides fragilis, B . vulgatus, B . splanchnicus, B . oralis, B . asaccharolyticus, B . melaninogenicus, Fusobacterium fusiforme (F . nucleatum), Sphaerophorus necrophorus, Clostridium perfringens, C . fallax, Propionibacterium acnes and several species of Peptococcaceae showed broth dilution MICs of 0.03 similar to 1 microgram/ml . Members of B . thetaiotaomicron, B . distasonis and S . freundii (F . mortiferum) were inhibited by 8 similar to 32 microgram/ml and 2 strains of S . varius had a broth dilution MIC of 256 microgram/ml . With most strains, the agar dilution MICs were 2 similar to 4 similar to 8 times the broth dilution MICs . In pour plate-tests, the MICs were not considerably influenced influenced by varying initial concentrations of viable cells . With most anaerobes, the MBCs of tiamulin were more than 100-fold higher than the MICs . The results obtained indicated that, apart from S . varius, B . thetaiotaomicron, B . distasonis and S . freundii (F . mortiferum), members of 16 other anaerobic species including B . fragilis were without exception sensitive to tiamulin.

Acta Pathol Microbiol Scand {B}, 1978 Aug, 86(4), 207 - 13
Analysis of amines and other bacterial products by head-space gas chromatography; Larsson L et al.; A gas chromatographic (GC) head-space technique is presented, which is suitable for the analysis of volatile products in bacterial broth cultures . This is exemplified by studies on Clostridium septicum, Klebsiella pneumoniae and Proteus mirabilis . The media were acidified or made alkaline and after heating, samples of the gas phase above the media were directly injected into the gas chromatograph . A gas chromatograph equipped with dual columns and flame ionization detectors was used, employing Porapak Q and Chromosorb 103 as stationary phases . Analysis of acidified media, using Porapak Q, gave chromatograms representing acidic and neutral volatile products, while when analysing samples made alkaline, using Chromosorb 103, alkaline and neutral compounds could be detected . Interest was particularly concentrated on the analysis of bacterial amines . P . mirabilis was found to produce isobutylamine and isopentylamine, which were identified by mass spectrometry and GC retention times C . septicum produced ethylamine . The GC head-space technique described constitutes a means for rapid identification of microorganisms . It is adaptable for use on a routine basis in the clinical microbiological laboratory.

Proc Natl Acad Sci U S A, 1978 Aug, 75(8), 3640 - 3
A rationale for stabilization of oxygen-labile enzymes: application to a clostridial hydrogenase; Klibanov AM et al.; A general procedure for stabilization of O2-labile enzymes exploiting "salting out" of oxygen from the microenvironment in the molecular layers immediately adjacent to charged surfaces of polyionic solid adsorbents has been developed . Empirical verification of this rationale is provided . The half-life of air inactivation of the O2-labile hydrogenase (EC 1.12.7.1) from Clostridium pasteurianum is increased 20- to 25-fold simply by adsorption (noncovalent binding) in dilute Tris.HCl buffer on common anion exchange supports such as DEAE-cellulose or Dowex 1-X2 . Predicted increases in degree of stabilization by using more densely charged adsorbents (such as polyethyleneimine-cellulose), as well as bulkier solvent counter-anions, are found; half-lives for air inactivation for the bound hydrogenase can be increased to 3000-fold longer than that of the free enzyme . Most of the total catalytic activity, assayed as H2 evolution from dithionite mediated by methyl viologen or ferredoxin, is retained, whereas the expected suppression of H2 uptake in the reverse reaction is observed.

Age Ageing, 1978 Aug, 7(3), 161 - 4
Age changes in collagen characteristics of bone and skin of a short-lived species of reptile; Panigrahy GK et al.; Decalcified bone collagen of older male garden lizards, Calotes versicolor, was less susceptible to digestion by collagenase from Clostridium histolyticum than that from younger individuals . In aged skin the percentage solubility and the soluble/insoluble collagen ratio decreased, with a concomitant rise in insoluble and total collagen . Collagen/unit area increased with advancing age in both dorsal and ventral skin . These results from a non-mammalian poikilothermic vertebrate provide additional evidence in favour of the cross-linkage theory of ageing and suggest a common pattern of collagen ageing in vertebrates.

J Clin Microbiol, 1978 Aug, 8(2), 238 - 41
Synergistic lysis of erythrocytes by Propionibacterium acnes; Choudhury TK; Sheep and human erythrocytes, partially processed by Staphylococcus aureus or Clostridium perfringens, were susceptible to lysis in the presence of Propionibacterium acnes . P . acnes liberated a lipase that was detected on Tween 80 agar and also on phospholipase C-precipitated egg yolk agar . Such a lipase might have contributed in the process of an intensified cellular lysis . Similar reactions were attempted with Lactobacillus acidophilus, known to possess a nondiffusible lipase, and failed to produce any such reactions . The synergistic reactions, between P . acnes and C . perfringens, were compared with The classical CAMP reaction in an attempt to find a correlation with the established membrane composition of the erythrocytes involved . Synergistic reactions observed do seem to reflect the membrane composition . Such findings, besides being contributory to an understanding of the role of these organisms in the process of pathogenesis, are of importance in the elucidation of molecular organization of biomembranes . Detailed studies, involving a large number of representative anaerobic bacteria, may also help provide an avenue in anaerobic species identification.

J Am Vet Med Assoc, 1978 Aug 1, 173(3), 306 - 7
Enterotoxemia in two foals; Dickie CW et al.; Two Quarter Horse foals from different premises died from enterotoxemia . Clostridium perfringens toxins alpha and beta were demonstrated in the foal's intestines by mouse protection tests . Clostridium perfringens type C was isolated from the intestines of each foal . Histologic examination revealed hemorrhage, necrosis, and massive numbers of C perfringens.

Infect Immun, 1978 Aug, 21(2), 678 - 80
Some properties of beta-toxin produced by Clostridium perfringens type C; Sakurai J et al.; Purified beta-toxin from Clostridium perfringens type C was found to be a single polypeptide chain protein with a molecular weight of approximately 30,000 . The toxin was heat labile, with 75% of its activity being inactivated by incubation at 50 degrees C for 5 min . Biological activity of the purified toxin was completely destroyed on exposure to trypsin for 30 min at 37 degrees C . The 50% lethal dose for mice was 1.87 microgram of purified toxin.

J Med Microbiol, 1978 Aug, 11(3), 299 - 302
In-vitro effects of Clostridium welchii type-D epsilon toxin on guinea-pig, mouse, rabbit and sheep cells; Buxton D; Epsilon toxin, at relatively low concentrations, killed guinea-pig peritoneal macrophages in vitro . The cells became swollen, the nuclear and cytoplasmic membranes "blistered" and discontinuous, and the cytoplasm appeared structureless . Formalinised epsilon prototoxin was shown to bind closely to the outer surface of the cells and it is concluded that this site represents the location of the receptors for epsilon toxin . In addition the toxin at higher concentrations killed rabbit peritoneal macrophages after increased periods of incubation, but had no demonstrable effect on other cells from guinea-pigs, rabbits, mice and sheep.

J Med Microbiol, 1978 Aug, 11(3), 289 - 92
The use of an immunoperoxidase technique to investigate by light and electron microscopy the sites of binding of Clostridium welchii type-D epsilon toxin in mice; Buxton D; Mice were given an intravenous dose of formalinised C . welchii type-D epsilon prototoxin and an immunoperoxidase technique was used to demonstrate this antigen in the tissues . The antigen was found to bind to the luminal surface of the endothelial lining of certain blood vessels, to the luminal surface of the cells lining the loops of Henle and distal convoluted tubules in the kidney, and to the hepatic sinusoids . As it has been shown previously that formalinised epsilon prototoxin and epsilon toxin can compete for the same receptor sites it is postulated that the binding sites demonstrated represent the location of the receptors for C . welchii type-D epsilon toxin.

J Med Microbiol, 1978 Aug, 11(3), 269 - 80
Neuraminidase production by clostridia; Fraser; The production of neuraminidase (EC 3.2.1.18) by a range of clostridial species was investigated with techniques previously developed to distinguish neuraminidase-negative and neuraminidase-positive strains of Clostridium perfringens (welchii) . Large amounts of extracellular neuraminidase were produced by representative strains of C . perfringens and C . septicum in the test media . Under similar conditions, two strains each of C . chauvoei and C . tertium were found to produce small amounts of the enzyme . All of 12 strains of C . sordellii were clearly shown to produce neuraminidase, often in large amounts, but none of five strains of the closely related but non-pathogenic C . bifermentans had demonstrable neuraminidase activity . No neuraminidase was produced by C . novyi (oedematiens) types A-D (10 strains), C . tetani (6), C . botulinum types A, B, C or E (4), C . sporogenes (4), C . histolyticum (4) or by single strains of five other clostridial species . Clostridial neuraminidase was predominantly extracellular and was not calcium-dependent . The investigation took account of variations in growth and enzyme production in different media . It was necessary to prolong the neuraminidase-assay reaction time to 24 h and to monitor for the presence of NAN-aldolase (EC 4.1.3.3) to define true negatives . It is suggested that neuraminidase production may be of value in taxonomic studies and that its production by several pathogenic species of clostridia may be of interest in studies of pathogenicity and virulence.

J Infect Dis, 1978 Aug, 138(2), 257 - 9
Effects of gentamicin on trypsin, chymotrypsin, and collagenase; Asch HL et al.; The effects of gentamicin on three proteolytic enzymes were studied . Gentamicin was tested at concentrations of 0.5-500 microgram/ml . Trypsin was tested at 0.5 microgram/ml using p-tosyl-L-arginine methyl ester and at 0.1 and 0.5 microgram/ml using azocoll as the substrate . Chymotrypsin was tested at 0.1 and 0.5 microgram/ml with azocoll . A soluble {14C}collagen assay was used to measure activity of collagenase derived from Clostridium histolyticum . The profiles of proteolytic activity vs . gentamicin concentration were similar for all three enzymes . At lower concentrations of gentamicin (less than 70 microgram/ml), there were two peaks of enhanced protease activity generally followed by inhibition . These unusual multiphasic effects of gentamicin on three different proteases are not presently understood, but they imply a previously unreported mode of action for this antibiotic.

Cancer Res, 1978 Aug, 38(8), 2295 - 300
Growth retardation and prevention of Ehrlich solid tumor by Clostridium perfringens type A spores and culture supernatant; Lapointe JR et al.; When given by direct s.c . injection into the Ehrlich solid carcinoma 1 week after s.c . tumor transfer, viable crude spores of Clostridium perfringens type A (attenuated mutant strain LNG11 ATCC 29348) inhibited tumor growth and significantly prolonged the life span of male outbred Swiss mice . Under these conditions a concentrated sterile supernatant of a C . perfringens culture proved to be slightly more effective than were viable crude spores . In contrast viable crude spores were ineffective in the treatment of female Swiss mice, but the sterile supernatant retained significant activity . When given at the time and site of s.c . grafting of Ehrlich tumor cells, a concentrated sterile supernatant of a C . perfringens culture prevented tumor growth in 80% of male outbred Swiss mice . Under these conditions viable crude spores prevented tumor growth in 70% of mice and significantly prolonged the life span in the other 30% . When given by i.p . injection and before i.p . grafting of tumor cells, viable crude spores of C . perfringens prevented Ehrlich ascites tumor in 5 of 12 Swiss mice and prolonged life span in the other 7 . In contrast concentrated sterile supernatant and viable purified spores were ineffective in the prevention or delay of the growth of Ehrlich ascites tumor . These data indicate that C . perfringens can be a potent antitumor agent without producing the harmful anaerobic infection of solid tumors (clostridial oncolysis.

J Med Microbiol, 1978 Aug, 11(3), 293 - 8
Further studies on the mode of action of Clostridium welchii type-D epsilon toxin; Buxton D; Intradermal injection of Clostridium welchii type-D epsilon toxin increased the permeability of blood vessels in guinea-pig skin to Evans blue dye by a mechanism not dependent on the release of histamine . The toxin was also found to raise the plasma concentration of cyclic adenosine 3', 5'-monophosphate in mice . It is concluded that epsilon toxin is an enterotoxin capable of causing widespread damage, after binding to receptor sites on the surface of certain cells, through a mechanism mediated by an adenyl cyclase-cAMP system.

Appl Environ Microbiol, 1978 Aug, 36(2), 386 - 8
Establishment of a heat inactivation curve for Clostridium botulinum 62A toxin in beef broth; Losikoff ME; A procedure is described for establishing a heat inactivation curve for the toxin of Clostridium botulinum 62A in beef broth . The effect of toxin titer, pH, and the type of acid employed for pH adjustment on the heat stability of the toxin is described.

Biochemistry, 1978 Jul 25, 17(15), 2943 - 8
Role of hydrophobicity in the binding of coenzymes . Appendix . Translational and rotational contribution to the free energy of dissociation; Janin J et al.; We calculate the loss of surface area accessible to solvent associated with coenzyme binding in Clostridium flavodoxin, in dogfish lactate dehydrogenase, and in lobster glyceraldehyde-3-phosphate dehydrogenase . The coenzymes are nearly buried in the complexes and lose on the order of 600 A2, while the proteins lose a similar amount of accessible surface area . Some of the loss can be attributed to conformation changes in the protein, at least in the case of lactate dehydrogenase, where we show that the apoenzyme has a larger accessible surface area than the holoenzyme . Using known correlations with the hydrophobic contribution to the free energy, we demonstrate that hydrophobicity is the major source of stabilization free energy in FMN binding to flavodoxin and in NAD binding to the two dehydrogenases: it contributes 25 to 30 kcal/mol to the free energy of dissociation, more than required in order to compensate for the loss of six degrees of translational/rotational freedom by the coenzyme.

Biochemistry, 1978 Jul 11, 17(14), 2857 - 63
Purification and characterization of a marine bacterial collagenase; Merkel JR et al.; A true collagenase was isolated from the culture fluid of a marine bacterium which has been designated Vibrio B-30 (ATCC 21250) . Collagenase production was obtained only in media containing collagen or certain degradation products of collagen . Partial purification on DEAE-cellulose and Sephadex G-200 columns produced active enzyme which was free of nonspecific proteases but which contained two collagenases . The two collagenases have the same apparent molecular size, and evidence is presented to support the theory that one collagenase is derived from the other . Vibrio B-30 collagenase appears to be a tetramer with a molecular weight of about 105 000 composed of two different subunits (mol wt 24 000 and 28 000) . Some of the properties of the Vibrio collagenase are compared with those of Clostridium histolyticum collagenase . Molecular weights, subunit structures, specificity and mode of collagen hydrolysis, insensitivity to diisopropyl fluorophosphate and calf serum, and sensitivity to certain metal ion complexing agents and isopropyl alcohol are similar for the collagenases from both organisms . However, Vibrio B-30 collagenase and Clostridium collagenase differ immunologically and electrophoretically.

J Biol Chem, 1978 Jul 10, 253(13), 4525 - 9
A pyruvate-containing peptide of proline reductase in Clostridium sticklandii; Seto B; Proline reductase in Clostridium sticklandii is composed of 10 apparently identical subunits . Each subunit contains a pyruvate residue that became labeled when the cell culture was supplemented with {14C}serine . No NH2-terminal amino acid was detected either by dansylation, by Edman degradation, or by aminopeptidase M digestion . The results suggest that the NH2 terminus may be blocked by pyruvate . A pyruvate-containing peptide, also blocked at the NH2 terminus, was isolated from the NH2-terminal portion of proline reductase . From amino acid analysis the peptide was found to be rich in basic amino acids and to have a molecular weight of 4621 . Its COOH-terminal amino acid was found to be serine and since the peptide was released from proline reductase by very mild alkali hydrolysis, it is suspected that an ester bond is involved.

Biochim Biophys Acta, 1978 Jul 7, 525(1), 45 - 54
The kinetics of methyl viologen oxidation and reduction by the hydrogenase from Clostridium pasteurianum; Erbes DL et al.; A mechanism for the reduction and oxidation of methyl viologen by Clostridium pasteurianum hydrogenase (hydrogen:ferredoxin oxidoreductase, EC 1.12.7.1) is proposed . Double reciprocal plots for methyl viologen reduction and oxidation at pH values 7.0-9.85 are linear, and the plots for reduction and oxidation are intersecting . Such data are consistent with a mechanism in which the H2 and one methyl viologen bind (either in order or randomly) with subsequent reduction and release of the methyl viologen . A second methyl viologen then is bound, reduced and released . Comparison of the calculated Keq' with the Haldane expression in which both methyl viologens react at the same rate show a large difference . This difference indicates that the two methyl viologens react at different rates . Addition of oxidized electron carriers inhibits the hydrogen-deuterium exchange reaction (i.e., the exchange of protons between H2 and 2H2O) . CO reversibly inhibits methyl viologen reduction and is competitive vs . H2 . O2 acts as an irreversible inhibitor.

Biochem J, 1978 Jul 1, 173(1), 129 - 44
Interaction between synthetic analogues of quinoxaline antibiotics and nucleic acids . Changes in mechanism and specificity related to structural alterations; Lee JS et al.; The interaction with DNA of six chemically synthesized derivatives of the quinoxaline antibiotics was investigated . Five of the compounds bound only weakly to DNA or not at all; for these substances spectrophotometric measurements, sedimentation studies with closed circular duplex bacteriophage-PM2 DNA and thermal-denaturation profiles were used to determine limits fot the binding constants . No interaction could be detected with two products of degradation of echinomycin (quinomycin A), one of which, echinomycinic acid dimethyl ester, had the lactone linkages opened, whereas the other retained an intact octapeptide ring but had a broken cross-bridge . The other compounds studied were des-N-tetramethyl-triostin A ('TANDEM') and its derivatives . A derivative of 'TANDEM' IN WHICH benzyloxycarbonyl moieties replace both quinoxaline chromophores had binding constants to nucelic acids in the range 10(2)--10(3)-1, whereas no interaction could be detected for a benzyloxycarbonyl derivative that, in addition, had the cross-bridge broken . The derivative of 'TANDEM' with L-serine in place of D-serine in both positions showed no detectable interaction with Clostridium perfringens DNA, whereas the binding constant to poly(dA-dT) was approx 2 X 10(3)M-1 . 'TANDEM' itself bound strongly to DNA, and the bathochromic and hypochromic shifts in its u.v.-absorption spectrum in the presence of DNA were similar to those seen with echinomycin . From the effect on the sedimentation coefficient of closed circular duplex bacteriophage-PM2 DNA the mechanism of binding was shown to involve bifunctional intercalation, typical of the naturally occurring quinoxaline antibiotics . Solvent-partition analysis was used to determine binding constants for the interaction between 'TANDEM' and a variety of natural and synthetic DNA species . The pattern of specificity thus revealed differed markedly from that previously found with the naturally occurring quinoxaline antibiotics . Most striking was the evident large preference for (A + T)-rich DNA species, in complete contrast with echinomycin and triostin A . The highest binding constant was found for poly(dA-dT), the interaction with which appeared highly co-operative in character . The conformations adopted by those quinoxaline compounds that bind strongly to DNA were examined withe aid of molecular models on the basis of results derived from n.m.r . and computer studies . It appears that the observed patterns of base-sequence specificity are determined, at least in part, by the structure and conformation of the sulphur-containing cross-bridge.

J Gen Microbiol, 1978 Jul, 107(1), 85 - 91
Proteases produced by a proteolytic mutant of Clostridium botulinum type E; Nakane A; A proteolytic mutant from Clostridium botulinum type E produced extracellular proteases after the end of exponential growth coinciding with the period of sporulation . Proteases were separated into four fractions by chromatography on a DEAE-cellulose column . One was a sulphydryl-dependent protease that also apparently required a divalent cation for enzyme activity since it was inhibited by EDTA . This enzyme hydrolysed synthetic amide and ester compounds containing an arginine residue, and showed some activity towards L-lysine methyl ester . It appeared that two of the other proteases were serine proteases and the fourth was a metal protease . These last three proteases did not require a thiol agent and did not hydrolyse any of the synthetic amides or esters examined . Only the sulphydryl-dependent protease could activate C . botulinum type B, E and F toxins . The ability of this enzyme to activate type B and E toxins was markedly lower than that of trypsin . The susceptibility of type B toxin to this protease was lower than that of type E toxin . C2 toxin was not activated by this enzyme . It is suggested that the sulphydryl-dependent protease in this proteolytic mutant of C . botulinum type E has properties similar to those of proteases from C . botulinum types B and F.

Infect Immun, 1978 Jul, 21(1), 59 - 63
Intraintestinal toxin in infant mice challenged intragastrically with Clostridium botulinum spores; Sugiyama H et al.; Conventionally raised suckling mice were injected intragastrically with 10(5) spores of a Clostridium botulinum type A culture . Botulism was not observed, but 80% or more of mice challenged when 8 to 11 days old had botulinum toxin in the large intestine 3 days later . Mice younger than 7 days or older than 15 days were resistant to the challenge . When in vivo toxin production was started by spores given to 9-day-old mice, toxin was present in the intestine at 1 through 7 days postchallenge but with greatest consistency between 1 and 4 days . Total toxin in an intestine ranged up to 1,920 50% lethal doses as titrated intraperitoneally in adult mice . The dose infecting 50% of a group of 9-day-old mice was 700 (95% confidence limits of 170 to 3,000) spores per animal . Toxin was formed in the lumen of the large intestine; it was not associated with the ileum . Injection of 10(5) spores intraperitoneally into 9-day-old mice resulted in toxin production in the large intestines of 30% of the test animals.

Appl Environ Microbiol, 1978 Jul, 36(1), 210 - 1
System for evaluating clostridial inhibition in cured meat products; Robach MC et al.; A method for evaluating inhibition of Clostridium botulinum, C . sporogenes, and C . perfringens in cured meat products was developed . This system can easily be used in the microbiology laboratory using aluminum ointment tubes as the product container . Swells caused by gas production by the organism are easily observed by using the aluminum tubes . Results obtained confirmed earlier work on the inhibitory effect of sodium nitrite and sorbic acid against the clostridia in cured meat products.

Can J Comp Med, 1978 Jul, 42(3), 357 - 63
Enterotoxigenic Clostridium perfringens type A isolated from intestinal contents of cattle, sheep and chickens; Niilo L; One hundred and fourteen strains of Clostridium perfringens, isolated from the intestinal contents of cattle, sheep, and chickens with enteritis or other disease conditions were studied for their ability to produce enterotoxin . Reversed passive hemagglutination, fluorescent antibody and immunodiffusion tests were used . On the basis of the reversed passive hemagglutination titres, supported by the other two tests, enterotoxigenicity of the strains was arbitrarily classified into two categories: highly enterotoxigenic and potentially enterotoxigenic, with 12% falling into each category . All the highly enterotoxigenic strains originated from cases of enteritis and included all three animal species . Apart from enterotoxigenicity, one C . perfringens strain produced beta toxin (type C) and 21 strains produced large amounts of alpha-toxin . The latter strains were predominantly associated with necrotic enteritis in chickens.

J Assoc Off Anal Chem, 1978 Jul, 61(4), 785 - 8
Method for maintaining viability of Clostridium perfringens in foods during shipment and storage: collaborative study; Harmon SM et al.; A collaborative study was conducted in 12 laboratories to determine the effectiveness of a new method for maintaining vegetative cells of Clostridium perfringens in viable condition during storage and transport of food specimens to the laboratory . The collaborative results showed that treatment of brown gravy and roast beef samples with an equal amount by weight of sterile buffered glycerol-sodium chloride solution to give a final 10% glycerol concentration and storage with Dry Ice for 10 days at -56 degrees C resulted in plate counts of C . perfringens which were 2-4 log cycles higher with 2 different strains than counts with untreated specimens stored by the usual method at -20 degrees C . Plate counts obtained with the treated specimens stored with Dry Ice were less than 1 log cycle lower than counts made with identical specimens before freezing for storage and shipment to the collaborators . The results with treated specimens were also more uniform among the different laboratories . Because the new method for storage and shipment of food samples was so effective for maintaining viability of the organism, the official first action method for C . perfringens (46.B01) was changed to incorporate these procedures as part of the method.

J Pharm Sci, 1978 Jul, 67(7), 900 - 5
Synthesis and quantitative structure-activity relationships of antibacterial 1-(substituted benzhydryl)-4-(5-nitro-2-furfurylideneamino) piperazines; Yung DK et al.; 1-Benzhydryl -4- (5-nitro-2-furfurylideneamino) piperazine and 11 substituted analogs were prepared and examined for in vitro antimicrobial activity . The compounds were active against Bacillus cereus 7, Bacillus megaterium 122, Bacillus subtilis 104, Clostridium perfringens 13, and the tetracycline-resistant Clostridium perfringens 37 . Regression analyses on the antibacterial activity data based on the Hansch approach, using pi, pi2, and sigma parameters, yielded several statistically significant correlation equations . 1-Benzhydryl-4-(5-nitro-2-furfurylideneamino) piperazine stopped the protein and DNA syntheses in C . perfringens 13, as indicated by precipitable radioactivity . The compound, however, showed no effect on the cell wall synthesis in the bacteria.

Mol Cell Biochem, 1978 Jun 15, 20(1), 25 - 40
Binding of protein chemotactic factors to the surfaces of neutrophil leukocytes and its modification with lipid-specific bacterial toxins; Wilkinson PC et al.; The binding to neutrophil leukoyctes of human serum albumin (HSA), which is chemokinetic for leukocytes, i.e . influences their rate of locomotion, and of alkali-denatured HSA, which is chemotactic for leukocytes, i.e . influences their direction of locomotion, was studied . Native serum albumin showed low affinity binding to the neutrophil surface . Denatured serum albumin showed saturable binding with a Ka of approximately 1-(6) litres per mole to about 10(6) binding sites per cell . Another protein chemotactic factor, alpha5-casein, gave similar binding . These results exclude that chemotactic reactions to denatured proteins are mediated in a completely non-specific manner and suggest the presence on the cell of a restricted number of defined recognition sites . Binding was reduced following treatment of the cells with either of two lipid-specific bacterial toxins, perfringolysin, the theta-toxin of Clostridium perfringens, an oxygen-labile cholesterol-specific toxin, and Staphylococcus aureus Sphingomyelinase C . Both have previously been shown to reduce chemotactic reactions and both were used at doses which did not reduce cell viability . These results suggest an important, and possiblly direct, role for membrane lipid in the binding sites for chemotactic factors . Visual analysis of the behaviour of perfringolysin-treated neutrophils showed that these cells were still capable of chemotactic locomotion . The cells appeared to be less efficient than normal in detecting chemotactic gradients only when at a distance from the gradient source, a finding which is consistent with reduced binding of the chemotactic factor to the cell surface.

Schweiz Med Wochenschr, 1978 Jun 10, 108(23), 847 - 53
{Gas gangrene . The files of the Swiss Accident Insurance Company 1963-1975}; Stamm B et al.; The clinical histories of 28 cases registered as gas gangrene by the Swiss National Accident Insurance (Schweizerische Unfallversicherungsanstalt) from 1963 to 1975 are reviewed . According to the classification of Altemeier (1 . gas gangrene, 2 . clostridial cellulitis, 3 . simple contamination of a wound by Clostridium, and 4 . gaseous infection without Clostridium) only 5 cases were assignable to group one (1 survivor), 2 cases to group two, 9 to group three and 9 to group four . Typical cases from each group are discussed to illustrate the advantages of this classification, the dangers of treatment based on wrong diagnosis, and the necessary prophylaxis . The more serious prognosis in gas gangrene in comparison to the other groups is emphasized.

J Biol Chem, 1978 Jun 10, 253(11), 4031 - 5
Characterization of gangliosides from bovine erythrocyte membranes; Chien JL et al.; Two glucosamine-containing gangliosides, sialosylhexaglycosylceramides, were isolated from bovine erythrocyte membranes . Both gangliosides were hydrolyzed by neuraminidase isolated from Clostridium perfringens to become neutral hexaglycosylceramides . Based on the results of sequential enzymatic hydrolysis and gas chromatography-mass spectrometric analyses of the methylated sugars, the structures of these two gangliosides were shown to be NeuAcalpha2 leads to 3Galbeta1 leads to 4GlcNAcbeta1 leads to 3Galbeta1 leads to 4GlcNAcbeta1 leads to 3Galbeta1 leads to 4Glc-ceramide and NeuGcalpha2 leads to 3Galbeta1 leads to 4GlcNAcbeta1 leads to 3Galbeta1 leads to 4GlcNAcbeta1 leads to 3Galbeta1 leads to 4Glc-ceramide, respectively . In addition, N-acetyl- and N-glycolylneuraminosyllacto-N-neotetraosylceramides, and N-acetyl- and N-glycolylneuraminosyllactosylceramides were also found in bovine erythrocytes . The predominant fatty acids in these two gangliosides were C 22:0 and C 24:0 . C-18 sphingosine was the major base detected.

Can J Microbiol, 1978 Jun, 24(6), 716 - 24
Stable isotope fractionation by Clostridium pasteurianum . 2 . Regulation of sulfite reductases by sulfur amino acids and their influence on sulfur isotope fractionation during SO32- and SO42- reduction; Laishley EJ et al.; In addition to an assimilatory sulfite reductase, studies of cultures of Clostridium pasteurianum supplemented with methionine, cysteine, and 35SO42- provides evidence for another reductase which is induced by SO32- . This inducible reductase appears to be dissimaltory because of the copious sulfide production arising when the cells are grown on SO32- . Cysteine can repress the assimilatory sulfite reductase but does not affect the inducible reductase . During late logarithmic growth on 1 mM SO42- + 10mM cysteine, depression of the inducible reductase occurred along with increased sulfide production . The presence of 1 mM cysteine and (or) 1 mM cysteine and (or) 1 mM methionine does not affect the inverse sulfur isotope effect for evolved H2S . However, 5 and 10 mM cysteine reduce the maximum delta34S value for released H2S from +40 to 10% . A small conversion of cysteine to H2S by C . pasteurianum occurs, but only in the stationary phase.

Bioinorg Chem, 1978 Jun, 8(6), 477 - 91
Role of chelation and water binding of calcium in dormancy and heat resistance of bacterial endospores; Rajan KS et al.; The possible relationship between the water binding by bacterial endospores and their dormancy and heat resistances has been examined in terms of the coordination characteristics of the spore-bound calcium . Stabilities of the calcium complexes of typical cytoplasmic and structural spore components were determined by potentiometric equilibrium pH measurements in model systems consisting of DPA, glycine, alanine, glutamic acid, alanyl-glutamic acid, triglycine, and tetraglycine . The Ca++-form and H+-form spores of Clostridium botulinum 33A were investigated in vivo with respect to their water sorption and heat-resistance characteristics . The results suggest that the complexing of calcium and Ca(II)-DPA may be biologically significant for spore resistance and dormancy at the following three levels: (1) complexing with spore cytoplasmic pool constituents consistent with the idea of a metal-chelate cross-linked cytoplasm or spore cement stabilizing the essential biological macromolecules, (2) complexing with structural components of the spore as indicated by the interaction with model peptides, and (3) coordination with water to produce an apparently dehydrated environment in the spore as evident from the much greater water-sorption capacity of the Ca++-form spores vs the much smaller water sorption of the H+-form spores . Interestingly enough, DPA itself, in the absence of metal ion, showed some interaction with di-, tri-, and tetrapeptides and a weak but detectable interaction with amino acids . Although the exact mode of the DPA-peptide interaction is not clear, it is attractive to speculate about its possible involvement in the control of spore dormancy and resistance.

J Hyg (Lond), 1978 Jun, 80(3), 431 - 8
Clostridium botulinum in aquatic environments in Great Britain and Ireland; Smith GR et al.; Mud samples from aquatic environments in many parts of Great Britain and Ireland were collected, mainly in 1975 and 1976, and examined for Clostridium botulinum . The samples were taken from lakes, ponds, reservoirs, marshes, mudflats, streams, rivers and canals, and the sampling areas included a number of bird refuges and reserves . Of 554 samples 194 (35.0%) were positive and 167 (30.1%), 19 (3.4%), 6 (1.1%) and 15 (2.7%) contained types B, C, D and E respectively; 13 (2.3%) contained more than one type . Each type demonstrated was found in both fresh-and salt-water environments . Type B was widespread; types C, D and E were demonstrated in widely separated areas in England and Wales, and type C was found in both the north and south of Scotland . The results were compared with those reported earlier in respect of surveys in the London area, the Norfolk Boads and the Camargue (France).

Zhonghua Min Guo Wei Sheng Wu Xue Za Zhi, 1978 Jun, 11(2), 50 - 61
Effect of metal ions on growth and sporulation of Clostridium perfringens in a synthetic medium; Lee KY et al.; All cells, whatever their origin, function in an essentially inorganic environment . In this environment, metal cations play an important role . Attempts were made in the present study to determine if different amounts of five metal ions (MG++, Fe++, Mn++, Zn++, Ca++) are needed for secondary metabolism of Clostridium perfringens than are needed for primary metabolism . Both the vegetative growth stage (primary metabolic stage) and spore stage (secondary metabolic stage) of Clostridium perfringens were studied . Endeavors were made to detect the effects of metal ions, if any, on growth and sporulation of the organisms . Mg++ was required for growth of vegetative cells and maintenance of normal cellular morphology, Fe++ was needed for sporulation as well as for vegetative growth, but the amount needed for spore formation was higher than that needed for growth . Ca++ was essential to refractility and heat-resistance of spores . Zn++ inhibited both growth and sporulation, if the Mg++ concentration was low . Mn++ was required for neither growth nor sporulation . For maximum heat-resistance of the spores, Ca++ plus unidentified organic substances were required.

Am J Vet Res, 1978 Jun, 39(6), 1049 - 51
Equine grass sickness: serologic evidence of association with Clostridium perfringens type A enterotoxin; Ochoa R et al.; Clostridium perfringens type A enterotoxin seroneutralization was carried out on sera from 50 horses recovered from grass sickness and from 100 other horses with no record of having had the disease . Of the affected horses, 70% had seroneutralizating titers higher than 1:64, half of these being equal or higher than 1:128 . More than 88% of the horses with no record of grass sickness had titers lower than 1:64 . These data support the theory of association between C perfringens type A toxins and grass sickness.

Can J Microbiol, 1978 Jun, 24(6), 643 - 9
Biological nitrogen fixation in the terrestrial environment of a high Arctic ecosystem (Truelove Lowland, Devon Island, N.W.T.); Jordan DC et al.; Arranged in descending order of nitrogen-fixing (acetylene-reducing) potential the sites examined were mesic meadow and peat polygon troughs (equal rank), transition zone between mesic meadow and gravel ridge, gravel ridge, polar dessert, and peat polygon tops . The dominant nitrogen-fixing microorganisms, as in other Arctic areas, were blue-green bacteria, especially those epiphytic on Arctic mosses . The epiphytic association exhibited an optimum temperature for fixation of 20 degrees C . Other bacteria potentially able to fix nitrogen were present in the soils examined but their activity was severely restricted by low soil temperatures and lack of readily utilizable energy sources . These bacteria included members of the genera Klebsiella (the most numerous), Bacillus, Clostridium, and Beijerinckia (scarce) . Also present at many of the sites was an unidentified yellow-pigmented fixer which was not Mycobacterium flavum . All fixers were psychotrophic rather than psychrophilic, having an optimum temperature greater than 20 degrees C but capable of slow growth at 5 degrees C or lower . The rate of acetylene reduction by the epiphytic system increased with the number of successive exposures to acetylene, a phenomenon of some significance in any calculations designed to measure the amount of nitrogen fixed in certain ecosystems.

Calcif Tissue Res, 1978 May 26, 25(2), 127 - 31
The interaction of collagenase with hydroxyapatite and related materials and enzymatic properties of the adsorbed enzyme; Knuuttila M et al.; The commercial collagenase from Clostridium histolyticum was adsorbed on hydroxyapatite, on bovine femur shaft and on enamel and dentin powders . The substrate specificity of the adsorbed enzyme tested, with chromophore substrate, azocoll and native collagen, differed from that obtained with the soluble enzyme . The adsorption and the substrate specificity was also dependent on the adsorbent used . A pretreatment of hydroxyapatite with chondroitin sulphate, hyaluronate and DNA lowered the adsorption of collagenase . Phosphate ion caused desorption of the enzyme from hydroxyapatite . Sodium fluoride caused partial desorption of the enzyme from hydroxyapatite and enamel and dentin powders . Colangenase adsorbed on root surfaces of teeth liberated hydroxyproline containing material.

Biochemistry, 1978 May 16, 17(10), 1866 - 72
Nitrogenase: the reaction between the Fe protein and bathophenanthrolinedisulfonate as a probe for interactions with MgATP; Ljones T et al.; The reaction between the Fe(II) chelating agent, bathophenanthrolinedisulfonate, and the iron-sulfur cluster in the Fe protein of nitrogenase from Clostridium pasteurianum has been studied . This reaction is greatly accelerated by the presence of MgATP . Analysis of the relationship between reaction rate and concentration of MgATP supports a model in which both of two binding sites for MgATP on the Fe protein must be occupied before the protein undergoes a conformational change, allowing the iron-sulfur site to react rapidly with chelator . This model is also consistent with presently available data on equilibrium binding of MgATP to the Fe protein . MgADP inhibits the effect of MgATP on the chelator reaction in a manner which suggests that MgADP binds strongly to one of the MgATP sites and more weakly to the other . Loss of enzymic activity due to exposure to O2 or 0 degrees C is accompanied by a decrease in the ATP-specific chelator reaction . Hence, this reaction was used to estimate the concentration of active iron-sulfur centers for the purpose of computing the extinction coefficient of the Fe protein, giving the value delta epsilon 430nm(ox-red) = 6600 M-1 cm-1.

Biochim Biophys Acta, 1978 May 11, 524(1), 188 - 97
Specific cleavage of reduced and S-carboxamidomethylated neurophysin II by the collagenase of Clostridium histolyticum; Katayama S et al.; Purified collagenase of Clostridium histolyticum was shown to cleave reduced and S-carboxamidomethylated bovine neurophysin between Cys-13 and Gly-14 . The scission resulted in formation of two separable fragments: a smaller peptide arising from residues 1 through 13, and a larger peptide comprising the remainder of the residues of the protein . By dansylation procedures, the smaller peptide was shown to have amino-terminal alanine as expected from the sequence of neurophysin II, and the larger peptide had amino-terminal glycine as anticipated . These results show that collagenase indeed cleaves bovine neurophysin II in accord with the specificity postulated for that enzyme, i.e., scission between -X-Gly- in a sequence of -Pro-X-Gly-Pro-Y- . This result, obtained with a non-collagenous protein substrate, is further confirmation of the specificity of collagenase as established by its action on collagens and on synthetic oligopeptides.

Infect Immun, 1978 May, 20(2), 526 - 9
Clindamycin-induced enterocolitis in hamsters as a model of pseudomembranous colitis in patients; Chang TW et al.; Stools from a patient with antibiotic-associated colitis and cecal contents from a hamster with clindamycin-induced enterocolitis were compared in a cytotoxicity assay to determine common properties . Both specimens produced actinomorphic changes in human amnion cells at 10(-7) dilutions . The toxin was acid labile, heat labile, nonether extractable, non-dialyzable, and produced maximum activity at 60% with ammonium sulfate precipitation . Cytotoxicity was neutralized with clostridial antitoxin but not with equine serum . Clostridium difficile was recovered in high concentrations in specimens from both the hamster and patients . The supernatants of these C . difficile strains produced cytoxic effects which were also neutralized by clostridial antitoxins . These results indicate that clindamycin-induced enterocolitis in hamsters is a model of human disease and implicate toxin-producing clostridia as responsible agents.

Appl Environ Microbiol, 1978 May, 35(5), 886 - 9
Causes of variation in botulinal inhibition in perishable canned cured meat; Tompkin RB et al.; Final internal processing temperatures within the range of 63 to 74 degrees C did not alter the degree of botulinal inhibition in inoculated perishable canned comminuted cured pork abused at 27 degrees C . Adding hemoglobin to the formulation reduced residual nitrite after processing and decreased botulinal inhibition . Different meats yielded different rates of botulinal outgrowth when substituted for fresh pork ham . Pork or beef heart meat showed no inhibition of the Clostridium botulinum inoculum even with a 156-microgram/g amount of sodium nitrite added to the product . This effect appears to be one of stimulating outgrowth, since residual nitrite depletion was not measurably altered.

Appl Environ Microbiol, 1978 May, 35(5), 863 - 6
Effect of prior refrigeration on botulinal outgrowth in perishable canned cured meat when temperature abused; Tompkin RB et al.; Perishable canned cured meat inoculated with Clostridium botulinum spores was placed at 4.4 or 10 degrees C after manufacture . Spore germination occurred at 10 degrees C . The germinated cell count remained stable over a period of 16 to 18 weeks . During that time period the inhibitory system and residual nitrite descreased . These factors combine to make perishable canned cured meats more prone to spoilage and potential hazard if they are temperature abused after a period of refrigerated storage.

Prikl Biokhim Mikrobiol, 1978 May-Jun, 14(3), 399 - 404
{Separation and purification of isoenzymes of Clostridium perfringens phospholipase C}; Shemanova GF et al.; The procedure for separation and purification of isoenzymes of phospholipase C from Clostridium perfringens (PLC) was developed . The procedure included primary concentration of culture liquid proteins and isoenzyme separtion on DEAE-cellulose during negative sorption of the major isoenzyme . Further purification of the isoenzymes was achieved by (NH4)2SO4 fractionation by Sephadex gel-filtration and isoelectric focusing . During polyacrylamide gel electrophoresis and precipitation reaction in the agar with the antiperfringens hyperimmune serum alpha1-PLC preparations were homogenous . Their coefficient of sedimentation was 3.8 S and isoelectric point was 5.50 . The minor isoenzyme alpha2-PLC had the same coefficient of sedimentation and the isoelectric point of 5.35.

Infect Immun, 1978 May, 20(2), 325 - 33
Effect of zinc and calcium ions on the production of alpha-toxin and proteases by Clostridium perfringens; Sato H et al.; Clostridium perfringens produced at least three distinct proteases in a synthetic medium containing calcium . Two of them, thiol and ethylenediaminetetraacetic acid disodium salt-sensitive proteases, appeared at an early stage of growth, but the other one, perhaps being identical to the one produced in a calcium-deficient medium, appeared at a late stage . The production of these proteases depended on Ca2+ but not on Zn2+ in the medium . Alpha-toxin, perhaps being a zinc-containing metalloenzyme, was rather resistant to the proteases, but toxin, produced in a zinc-deficient medium or deprived of zinc with ethylenediaminetetraacetic acid disodium salt, was very sensitive . By adding Zn2+, the toxin lacking zinc may have been converted to the zinc-containing metalloprotein that is resistant to proteases . This may explain why alpha-toxin activity increased progressively in a zinc-containing medium in spite of simultaneous production of potent proteases and why it disappeared rapidly in a zinc-deficient medium.

Can J Microbiol, 1978 May, 24(5), 633 - 5
Efficacy of laboratory tests for the detection of enterotoxigenic Clostridium perfringens; Niilo L; Nineteen Clostridium perfringens strains with positive erythemal and ligated intestinal loop reactions, and 22 strains with negative reactions, originating from food-poisoning cases, were tested comparatively using the fluorescent antibody (FA), reversed passive hemagglutination (RPHA), and immunodiffusion (ID) tests . All the biologically positive strains were detected by the three immunological tests used . The FA test detected five additional strains among the biologically negative group which did not react in RPHA or ID tests . Sporulating culture supernatant fluids, after 13 to 17h of growth, were satisfactory for testing for the presence of enterotoxin by the RPHA and ID tests . The FA test was used on cell smears.

J Bacteriol, 1978 May, 134(2), 597 - 605
Molybdenum independence of nitrogenase component synthesis in the non-heterocystous cyanobacterium Plectonema; Nagatani HH et al.; The cyanobacterium Plectonema boryanum (IU 594-UTEX 594) fixes N2 only in the absence of combined N and of O2 . We induced nitrogenase by transfer to anaerobic N-free medium and studied the effect of Mo starvation on nitrogenase activity and synthesis . Activity was first detected within 3 h after transfer by the acetylene reduction assay in controls, increasing for at least 25 h . Cells grown on nitrate and Mo and then transferred to N-free, Mo-free medium produced 8% of the control nitrogenase activity . Addition of W to the Mo-free medium reduced the activity to 0.5% . Under both Mo starvation conditions, nitrogenase protein components were synthesized . Component II of the cyanobacterial enzyme was detected by in vitro complementation with Mo-containing component I from Klebsiella pneumoniae or Azotobacter vinelandii but not Clostridium pasteurianum . Component I activity was restored by addition of Mo to cultures in which new enzyme synthesis was blocked by chloramphenicol . Acidified extracts of Plectonema induced in Mo-containing medium contained the Fe-Mo cofactor required to activate extracts of the Azotobacter mutant UW45 in vitro, but they did not activate extracts of Mo-starved Plectonema . Analysis of 35SO4(2-)-labeled proteins by polyacrylamide gel electrophoresis suggested that Mo is required for the conversion of a high-molecular-weight precursor to component I in Plectonema.

Arch Microbiol, 1978 Apr 27, 117(1), 53 - 60
Regulatory properties of the nitrogenase from Rhodopseudomonas palustris; Zumft WG et al.; Ammonium salts, glutamine, asparagine, and urea cause an immediate inactivation (switch-off) of light-dependent acetylene reduction in intact cells of the photosynthetic bacterium Rhodopseudomonas palustris . This effect is reversible showing the same kinetic pattern of inactivation and reactivation with all effector compounds . Its duration depends on the amount of effector added to the cells . Both nitrogenase components are found catalytically active in a cell-free preparation after enzyme switch-off in vivo . Involvement of the ammonia assimilating system in this regulatory mechanism is indicated by the following observations: ammonia uptake during the switch-off period, resumption of acetylene reduction after disappearance of ammonia from the outer medium, and persistence of enzyme switch-off with dihydrogen and thiosulfate as electron donors in the absence of an additional carbon source . Nitrogenase activity in crude extracts is non-linear with time and is stimulated by manganese ions . After resolution of nitrogenase into its MoFe--protein and Fe--protein these properties are lost, indicating the presence of an activating factor . Nitrogenase of R . palustris cross reacts reciprocally with the complementary proteins of Azotobacter vinelandii, but not with those of Clostridium pasteurianum.

Biochim Biophys Acta, 1978 Apr 20, 508(3), 464 - 77
Role of negatively charged phospholipids in highly purified (Na+ + K+)-ATPase from rabbit kidney outer medulla studies on (Na+ + K+)-activated ATPase, XXXIX; de Pont JJ et al.; 1 . The requirement for specific polar head groups of phospholipids for activity of purified (Na+ + K+)ATPase from rabbit kidney outer medulla has been investigated . 2 . Comparison of content and composition of phospholipids in microsomes and the purified enzyme indicates that purification leads to an increase in the phospholipid/protein ratio and in phosphatidylserine content . 3 . The purified preparation contains 267 molecules phospholipid per molecule (Na+ + K+)-ATPase, viz . 95 phosphatidylcholine, 74 phosphatidylethanolamine, 48 spingomyelin, 35 phosphatidylserine and 15 phosphatidylinositol . 4 . Complete conversion of phosphatidylserine into phosphatidylethanolamine by the enzyme phosphatidylserine decarboxylase has no effect on the (Na+ + K+)-ATPase activity of the purified preparation . 5 . Complete hydrolysis of phosphatidylinositol by a phospholipase C from Staphylococcus aureus, which is specific for this phospholipid, has no effect on the (Na+ + K+)-ATPase activity . 6 . Hydrolysis of 95% of the phosphatidylcholine and 60--70% of the spingomyelin and phosphatidylethanolamine by another phospholipase C (Clostridium welchii) lowers the (Na+ + K+)-ATPase activity by about 20% . 7 . Combination of the phospholipid-converting enzymes has the same effect as can be calculated from the effects of the enzymes separately . Only complete conversion of both phosphatidylserine and phosphatidylinositol results in a loss of 44% of the (NA+ + K+)-ATPase activity and 36% of the potassium 4-nitrophenylphosphatase activity . 8 . These experiments indicate that there is no absolute requirement for one of the polar head groups, although in the absence of negative charges the activity is lower than in their presence.

Appl Environ Microbiol, 1978 Apr, 35(4), 782 - 90
Bacteria isolated from the duodenum, ileum, and cecum of young chicks; Salanitro JP et al.; Facultatively anaerobic and strictly anaerobic bacteria colonizing the intestinal tracts of 14-day-old chicks fed a corn-based diet were enumerated, isolated, and identified . Colony counts from anaerobic roll tubes (rumen fluid medium) or aerobic plates (brain heart infusion agar) recovered from homogenates of the duodenum, upper and lower ileum, and cecum varied appreciably among samples from individual birds . Anaerobic and aerobic counts from the duodenum and ileum were similar . Anaerobic counts were highest from the cecum (0.7 X 10(11) to 1.6 X 10(11)/g of dry tissue) and exceeded aerobic plate counts by a factor of at least 10(2) . Facultatively anaerobic groups (Streptococcus, Staphylococcus, Lactobacillus, and Escherichia coli) comprised the predominant flora of the duodenum and ileum, although large numbers of anaerobes (9 to 39% of the small intestine isolates), represented by species of Eubacterium, Propionibacterium, Clostridium, Gemmiger, and Fusobacterium, were also recovered . Strict anaerobes (anaerobic gram-positive cocci, Eubacterium, Clostridium Gemmiger, Fusobacterium, and Bacteriodes) made up nearly the entire microbial population of the cecum . Scanning electron microscopy of the intestinal epithelia of chicks revealed populations of microbes on the duodenal, ileal, and cecal mucosal surfaces.

J Hyg (Lond), 1978 Apr, 80(2), 175 - 81
Bacterial contamination in a modern operating suite . 4 . Bacterial contamination of clothes worn in the suite; Hambraeus A et al.; Clean clothes in the staff dressing rooms were heavily contaminated with bacteria, mainly Bacillus sp., but Staphylococcus aureus were found on 14% and Clostridium sp . on 10% of the garments examined . A comparison of the occurrence of Staph . aureus on shirts worn by staff in wards and operating departments showed ward shirts to be contaminated more heavily and with more strains . Examination of sterile gowns worn by surgeons showed that 70% were contaminated with Staph . aureus after operation . Of the strains isolated 31% were identical with those carried by the surgeon or by the patient operated on, but for the remainder no source could be found.

Zh Mikrobiol Epidemiol Immunobiol, 1978 Apr, (4), 116 - 9
{Characteristics of spore formation in Clostridium botulinum type C}; Shakhbanov AA; Electron microscope study of C1 . botulinum, tyep C, showed that microbial cells were surrounded with a five-layer wall . Structures characteristic of sporulating cells and phage particles whose intracellular development led to reduction and lysis of the cytoplasm were revelaed in the area of the cytoplasm . Mature spores were encountered rarely . Formation of prespore, cortex was observed, but the elements of the spore membrane were chaotically dispersed in the whole cytoplasm . Such disturbances could be connected with the presence of phage in the culture.

Health Lab Sci, 1978 Apr, 15(2), 74 - 80
The occurrence of Clostridium botulinum and Clostridium tetani in the soil of the United States; Smith LD; Soil samples taken every fifty miles on four east-west transects across the United States were examined for C . botulinum and C . tetani, organisms that could inhibit the growth of C . botulinum type A, and for various soil properties . Type A strains were found mostly in the western part of the United States, in neutral to alkaline soil . Type B strains were more uniformly distributed, with a majority of them occurring east of the Mississippi River; none, however, were found in samples taken in the southermost transect . They were associated with soil of high organic content . Type C strains were found only in acid soil of the Gulf Coast, and type D strains in alkaline soil of some western states . Type E strains were mostly associated with damp to wet soil . Organisms inhibitory to type A strains were found in 4 of 21 samples of soil in which type A strains were demonstrated and in 7 of 20 samples in which they were not . Trypsin activation of culture fluids was necessary for the demonstration of most strains of types B, C, D, and E . C . tetani was demonstrated in 30 per cent of the soil samples . Its occurrence was not correlated with any particular soil type or climatic area.

Aust J Exp Biol Med Sci, 1978 Apr, 56(2), 139 - 45
Isolation of bacteriophage-like particles from uninduced Clostridium tetani cultures; Brown KJ et al.; The isolation of hexagonal-headed, tailless, bacteriophage-like particles from uninduced cultures of Clostridium tetani is described . Clear, round, 1--3 mm diameter plaques were noted on Clostrisel agar plates, which were overlaid with soft agar inoculated with 7--14 day broth cultures . Particles were detected by transmission electron microscopy from broth cultures seeded with scrapings from the plaques . Both electron dense and electron lucent heads were noted . An electron dense head was observed attached to the surface of a dividing bacterium.

Appl Environ Microbiol, 1978 Apr, 35(4), 762 - 5
Use of ethanol for selective isolation of sporeforming microorganisms; Koransky JR et al.; When mixed cultures containing sporeforming bacteria were treated with heat or with ethanol, the latter consistently resulted in better recovery of Clostridium and Bacillus species . Both techniques were effective in eliminating vegetative cells . An ethanol concentration greater than 25% and exposure for 45 min or longer were necessary to kill all vegetative cells in mixed-culture samples . Ethanol treatment (50% ethanol for 1 h) was effective for isolating sporeforming bacteria from intestinal specimens . Seven different species of Clostridium were the only bacteria isolated from the ethanol-treated specimen of intestinal contents from the large bowel of a patient . It was concluded that treatment with ethanol for 1 h is an effective technique for selective isolation of sporeforming bacteria from mixed cultures and certain types of clinical specimens.

J Hyg (Lond), 1978 Apr, 80(2), 267 - 74
Prophylaxis against tetanus in non-immune patients with wounds: the role of antibiotics and of human antitetanus globulin; Lowbury EJ et al.; The potential value of oral erythromycin for antitetanus prophylaxis in non-immune patients with open wounds was assessed . Serum obtained by venepuncture from health persons 2 h after an oral dose of an erythromycin preparation was used as a culture medium rendered anaerobic by addition of cooked meat . Strains of Clostridium tetani inoculated into these sera failed to multiply when the donor had taken 500 mg of erythromycin estolate before a meal; other erythromycin preparations and the estolate at a dosage of 250 mg were ineffective or inconsistent in their inhibition of the growth of Cl . tetani . Human antitetanus globulin (ATG) was given to 12 patients, 9 with severe injuries and 3 with extensive burns, all of whom were judged, from their history, to be non-immune (or with expired immunity); all except one had received large intravenous infusions of blood and/or other fluids . Serum antitoxin assays by a mouse protection technique on days 0, 1--2, 3--5, 6--10 and 14+ showed no detectable antitoxin (less than 0.01) unit/ml) in the initial (pre-ATG) sample from three patients with severe injuries and in one with extensive burns . All the patients in the severely injured group showed an early appearance or increase in tetanus antitoxin to protective titres . Two of the three severely burned patients showed, respectively, a delayed appearance or an increase in antitoxin; the other burned patients showed a reduction from the initial pre-ATG titre, followed by a return to that titre after day 5.

Zentralbl Bakteriol {Orig A}, 1978 Apr, 240(2), 221 - 34
{Use of the reversed passive hemagglutination in detection of Clostridium botulinum type A, B, and E toxin (author's transl)}; Sonnenschein B; With the reversed passive hemagglutination technique it is possible to detect minimal amounts of botulinum type A, B and E toxins (s . Tab . 3) . The antisera are used were prepared by foot--pad injection of rabbits with purified toxoids in Freund's complete adjuvant (s . Tab . 1) . Antitoxin globulin were prepared from rabbit antisera with (NH4)2SO4 to 50% . Formalinized and tanned human erythrozytes were sensitized with these specific antitoxin globulins . Only slight cross reactions ere encountered between the type A, B, and E antiglobulin sensitized cells and culture filtrates of C . butyricum, C . sporogenes (type A and B antiglobulin only) and C . perfringens type C (s . Tab . 4).

Appl Environ Microbiol, 1978 Apr, 35(4), 809 - 10
New modification of Willis and Hobbs' method for identification of Clostridium perfringens; Serrano AM et al.; A modification of the medium of Willis and Hobbs is described . All strains of Clostridium perfringens grown on it gave a positive lecithinase reaction . Some gave a negative reaction in the origin medium.

J Clin Pathol, 1978 Apr, 31(4), 359 - 60
Free-sporing Cl . welchii in ordinary laboratory media and conditions; Rahman M; A strain of Clostridium welchii produced spores in ordinary blood agar plates . Investigations confirmed that it was the character of this particular strain and that the laboratory media were not inducing sporulation . During a period of 12 months a total of 100 strains of Cl . welchii were studied . None of them produced spores in ordinary laboratory media and conditions when examined microscopically.

Zentralbl Bakteriol {Orig A}, 1978 Apr, 240(2), 215 - 20
New strains of Clostridium botulinum subtype Af; Gimenez DF et al.; Two new strains of Clostridium botulinum subtype Af have been isolated from soil samples of Mendoza Province, Argentina . Some serological and other biological properties of these new isolates have been studied in comparison with the prototype 84-SC2 strain . No differences in morphology and biochemical activities were found among these three so far known strains of this subtype . Neither serologic differences have been recorded, suggesting that the bispecificity and the type A to type F serologic ratio (A:F) is a stable genetic trait of subtype Af . The three strains were found in well separated areas . None of the type A strains isolated either from soil samples or from human botulism outbreaks in Mendoza have shown such a high degree of toxigenicity as that shown by Af strains . All these facts suggest that this peculiar type is not the product of a recent genetic relationship between types A and F . Rather, it represents a stable type in nature.

J Bacteriol, 1978 Apr, 134(1), 139 - 46
Identity of proline dehydrogenase and delta1-pyrroline-5-carboxylic acid reductase in Clostridium sporogenes; Costilow RN et al.; Proline dehydrogenase and delta1-pyrroline-5-carboxylic acid (PCA) reductase activities were copurified 60- and 130-fold, respectively, from extracts of Clostridium sporogenes . The primary change in the ratio of activites was the result of a loss of proline dehydrogenase activity during dialysis . Both activities were eluted in single peaks from diethylaminoethyl-cellulose, hydroxylapatite, and Sephadex G-200 columns . They had identical sedimentation coefficients (10.3S), as determined in linear sucrose gradients, and identical isoelectric points (4.95 to 5.12) based on isoelectric focusing . The proline dehydrogenase activity was dependent on nicotinamide adenine dinucleotide and L-proline, and the PCA reductase required L-PCA and reduced nicotinamide adenine dinucleotide . The optimum pH for the assay of proline dehydrogenase was approximately 10.2, whereas that for PCA reductase was 6.5 to 7.5 . An increase in pH from 8.0 to 10.2 greatly decreased the apparent Michaelis constant observed for L-proline, and an increase from pH 8.3 to 8.6 resulted in a large shift in the reaction equilibrium toward PCA . Both the dehydrogenase and reductase activities were stabilized to heating at 65 degrees C for 5 min by solutes of high ionic strength and were inactivated in a similar fashion when dissolved in low-ionic-strength buffer . The specific activities for both were reduced by about 50% when glucose was added to the growth medium . The data support the conclusion that L-proline and L-PCA are interconverted by either a single enzyme or an enzyme complex in extracts of C . sporogenes cells.

Tijdschr Diergeneeskd, 1978 Mar 15, 103(6), 303 - 11
{Food poisoning in cattle caused by ingestion of brewers' grains contaminated with Clostridium botulinum type B (author's transl)}; Breukink HJ et al.; In 1977, outbreaks of disease associated with serious losses occurred on twenty cattle farms . This disease was due to the fact that the animals had been fed brewers' grains . The clinical picture was marked by anorexia, profuse salivation, regurgitation and dehydration . The course of the disease varied with the severity of the attack . Toxicological tests were negative . Cl . botulinum type B as well as toxin type B were found to be present in the brewers' grains fairly often . As a rule, Cl . botulinum was also detected in the rumen contents and faeces . The treatment of the animals is described; this resulted in recovery in a number of cases . When experimental animals were fed brewers' grains from farms on which outbreaks had occurred, this induced similar clinical pictures . The results of studies on the aetiology and pathogenesis are discussed . It is concluded that the clinical picture was an atypical form of botulism caused by Cl . botulinum type B . Possible causes of this abnormal clinical picture are discussed in greater detail.

Arch Microbiol, 1978 Mar, 116(3), 253 - 7
Metabolic products of microorganisms . 170 . On the antibiotic activity of cladosporin; Anke H et al.; Cladosporin was isolated from the cultures of three species of the genus Eurotium . Cladosporin inhibited the growth of several fungi and at very low concentrations the growth of Bacillus brevis and Clostridium pasteurianum . Bacillus subtilis and most other Gram-positive bacteria were not sensitive . Gram-negative bacteria and yeasts were not affected by concentrations up to 100 microgram/ml . Dimethyl cladosporin showed only week activity against Bacillus brevis with the minimal inhibitory concentrations being a 100 times higher than of cladosporin . The incorporation of leucine and uracil into acid insoluble material in Bacillus brevis cells was completely inhibited by concentration of 0.5 microgram/ml cladosporin . The incorporation of thymidine was not affected at this concentration.

Infect Immun, 1978 Mar, 19(3), 1101 - 3
Inhibition of hemolysis by zinc and its reversal by L-histidine; Avigad LS et al.; Hemolysis induced by staphylococcal alpha-toxin, staphylococcal beta-toxin, streptolysin O, and streptolysin S was inhibited by zinc ions by virtue of inhibition of an early step in the events leading to lysis, presumably by preventing the lysins from attaching to the plasma membrane . In contrast, in hemolysis induced by Clostridium perfringens alpha-toxin and by perfringolysin O, a later step was inhibited by zinc . In hemolysis caused by saponin, lysolecithin, and Triton X-100, hemoglobin was precipitated by zinc ions as it was released from the erythrocyte . Inhibition by zinc was abolished by several amino acids of which L-histidine was the most effective.

Hoppe Seylers Z Physiol Chem, 1978 Mar, 359(3), 399 - 406
On the significance of sialic acid in high affinity 5-hydroxytryptamine uptake by synaptosomes; Dette GA et al.; Synaptosomes isolated from rat brain cortex incorporated {14C}5-hydroxytryptamine at 37 degrees C with high affinity . An apparent transport constant of Kt = 50nM was found . The high affinity uptake was decreased by treatment of synaptosomes with neuraminidase from Vibrio cholerae or Clostridium perfringens prior to incubation with {14C}5-hydroxytryptamine . The inhibition was related to the amount of sialic acid released, with a Ki value of 3.5 micrometer . A non-competitive type of inhibition was observed after treatment with neuraminidase . The inhibition caused by ouabain could not be enhanced by simultaneous treatment with neuraminidase . Neuraminidase did not lower the activity of (Na + K)-ATPase or Mg2-ATPase . These results suggest that sialic acid is involved in the 5-hydroxytryptamine uptake mechanism without functional linkage to the energy pump of the membrane, which maintains the sodium gradient necessary for 5-hydroxytryptamine transport.

Rev Can Biol, 1978 Mar, 37(1), 43 - 6
{Phage-like particles produced by Clostridium tetani}; Ackermann HW et al.; Two C . tetani strains used for toxin production spontaneously produce two varieties of phage-like particles with isometric heads . Type A has a contractile tail, whereas type B shows a non-contractile tail with a long, wavy tail fiber . No relationship between the presence of these particles and the amount of toxin produced was found.

Zh Mikrobiol Epidemiol Immunobiol, 1978 Mar, (3), 112 - 4
{Ultrastructure of Clostridium tetani . I . Structural features of the surface structures of vegetative cells}; Afanas'eva GA et al.; Crest-like structures formed by internal layer of cell wall and cytoplasmic membrane were revealed in G1.tetani 471 by electron microscopy with the use of negative contrasting, ultrathin sections and freezing-etching . The transverse section of these crest-like structures was 56.3 nm and they were localized 4 to 6 in one row girdling the protoplast in different directions . Ring-like subunits located in rows with the periodicity of 5.9 nm, perpendicularly to the long axis of the cell, were revealed on the surface of the cell wall.

Appl Environ Microbiol, 1978 Mar, 35(3), 533 - 9
Relation between radiation resistance and salt sensitivity of spores of five strains of Clostridium botulinum types A, B, and E; Kiss I et al.; The NaCl tolerance of different strains of Clostridium botulinum varies over a wide range, and the patterns of NaCl inhibition differ distinctly and characteristically from strain to strain . The more radiation-resistant strains, such as 33A, 62A, and 7272A, are more resistant to NaCl, whereas the more radiation-sensitive strains, such as 51B and 1304E, are more sensitive to NaCl . This rule appears to hold irrespective of whether the spores were unirradiated controls or whether they were radiation damaged prior to exposure to NaCl in the recovery media . The data seem to indicate that radiation doses in the shoulder portion of the radiation survival curves did not noticeably sensitive the spores to NaCl, whereas radiation doses in the exponential-decline portion of the survival curve invariably produced a distinct sensitization . Thus, strains 33A and 62A were not sensitized to NaCl by 0.3 to 0.4 Mrad, i.e., in the shoulder portion of the survival curve . Radiation-sensitive strain 51B, which shows no distinct shoulder in its survival curve, was sensitized to NaCl by 0.1 Mrad, the lowest radiation dose employed in this study . These observations seem to suggest a possible relationship between deoxyribonucleic acid repair capacity and salt tolerance.

J Clin Microbiol, 1978 Mar, 7(3), 307 - 9
Bacteriocin typing of Clostridium perfringens in human feces; Mahony DE et al.; Three hundred and ninety-nine isolates of Clostridium perfringens from enriched stool specimens of 51 individuals (about eight colonies per person) were typed by bacteriocins . Forty-nine percent of these persons carried more than one bacteriocin type in their stool, and some had three or four different stains as determined by bacteriocin typing . Weekly stool specimens obtained from seven positive volunteers over a period of 5 weeks were screened for C . perfringens, and several colonies from each person were typed . This survey demonstrated that the number of types fluctuated with time, several types could be carried simultaneously, and the isolation of the organism was variable . Nine new bacteriocin types of C . perfringens were isolated in this study.

Carbohydr Res, 1978 Mar, 61, 447 - 55
2-acetamido-2-deoxy-alpha-D-galactosidases of Clostridium perfringens . Separation and characterization of an exoglycosidase and an oligosaccharidase; Bell WC et al.; Two distinct 2-acetamido-2-deoxy-alpha-D-galactosidases have been separated from filtrates of cultured Clostridium perfringens by electrophoresis in 6.5% poly(acrylamide) gels . One of the enzymes had a mobility of 0.32--0.36 (relative to Bromophenol Blue) and was identified as the exoglycosidase, 2-acetamido-2-deoxy-alpha-D-galactosidase . It appears to be the same enzyme as that reported in 1972 by McGuire et al . The second of the two enzymes, having a relative mobility of 0.42--0.46, corresponds to the oligosaccharidase reported in 1972 by Huang and Aminoff . The A-specificities of human type-A erythrocytes and of water-soluble glycoproteins having A-activity are both destroyed by incubation with the 2-acetamido-2-deoxy-alpha-D-galactosidase, but not on incubation with the oligosaccharidase . A concomitant rise in blood-group O(H) activity, as indicated by the use of a lectin from Ulex europeus, occurred in the A-erythrocytes treated with the exoglycosidase 2-acetamido-2-deoxy-alpha-D-galactosidase.

Arch Intern Med, 1978 Mar, 138(3), 393 - 5
Anaerobic septicemia after transrectal prostatic biopsy; Harris LF et al.; Transrectal biopsy of the prostate resulted in anaerobic septicemia in two patients, despite parenteral gentamicin sulfate prophylaxis . Bacteroides fragilis sepsis developed subacutely in one patient having a postbiopsy pelvic abscess . Clostridium perfringens sepsis occurred fulminantly in another patient 24 hours after biopsy of a gland extensively involved with adenocarcinoma . These cases indicate a potential hazard of sepsis due to anaerobic contamination with rectal microflora at the time of transrectal prostatic biopsy and the futility of prophylaxis directed only at aerobic bacteria.

Eur J Biochem, 1978 Feb, 83(2), 375 - 85
A circular dichroism study of DNA.platinum complexes . Differentiation between monofunctional, cis-bidentate and trans-bidentate platinum fixation on a series of DNAs; Macquet JP et al.; The circular dichroism (CD) spectra of a series of DNA . platinum complexes are presented . The following platinum compounds, {Pt(dien)Cl}Cl, cis-Pt(NH3)2Cl2, cis-Pt(en)Cl2, trans-Pt-(NH3)2Cl2, K{Pt(NH3)Cl3} and K2{PtCl4} were complexed with the DNA extracted from bacteria Micrococcus lysodeikticus (72% dG + dC), Escherichia coli (50% dG + dC), Clostridium perfringens (32% dG + dC) and salmon sperm (41% dG + dC) . Strong differences were found between the different DNA . Pt complexes . Three types of spectra clearly demonstrate the different platinum binding modes on DNA . In the first type, the platinum compound, i.e . {Pt(dien)Cl}Cl, is fixed to DNA with only one bond (monofunctional complex formation) and no significant change of the CD positive band of DNA is found . The main feature of the second type is a continuous intensity decrease of the positive band as observed for trans-Pt(NH3)2Cl2 (trans-bidentate complex formation) . The third type concerns the cis-bidentate platinum fixation obtained with cis-Pt(NH3)2Cl2, cis-Pt(en)Cl2, K{Pt(NH3)Cl3} and K2{PtCl4} . The CD spectra are in this case characterized by an increase in the positive Cotton effect which is dG + dC-dependent up to an rb value around 0.10 (where rb = number of platinum atoms bound per nucleotide), followed by a decrease until DNA saturation with platinum is reached . A linear decrease in the amplitude of the negative band is detected in all the complexes except in the case of the monofunctional DNA . Pt complexes . For the cis-bidentate and trans-bidentate platinum fixation, a continuous bathochromic shift occurs.

Br J Surg, 1978 Feb, 65(2), 101 - 5
Gallstone disease presenting as septicaemic shock; Faber RG et al.; Three patients are described who were admitted to hospital with septicaemic shock due to gallstones, but in none of whom had the presence of gallstones, been diagnosed previously . All 3 had positive blood cultures, Clostridium welchii being isolated in one patient (in another patient this organism was isolated from the bile) . Two patients had renal failure requiring dialysis . All 3 patients survived . The management of this serious presentation of gallstones, hitherto poorly described in British literature, is discussed.

J Hyg (Lond), 1978 Feb, 80(1), 57 - 67
Bacterial contamination in a modern operating suite, 2 . Effect of a zoning system on contamination of floors and other surfaces; Hambraeus A et al.; In this investigation the bacterial contamination of surfaces such as walls and floors in a modern operating suite, together with surfaces of lamps in the operating theatres, and the clogs worn by staff, was studied . Counts of colony-forming units were made on impression plates containing blood-agar with Tween 80 for total bacterial counts, Baird Parker medium with egg yolk and tellurite for Staphylococcus aureus and trypticase peptone agar with neomycin and polymyxin for Clostridium spp . The areas examined were divided into the patients' route to the operating theatre, the staff's route, and the central area containing the operating rooms, anaesthetic rooms, and exit and scrub-up areas . In the patients' route counts of total organisms ranged from about 10000 to 30000/m2; for Staph . aureus the range was from 70 to 540/m2 . In the staff's route the highest count was about 70000/m2 in the dressing area, and the numbers of Staph . aureus were about the same as along the patients' route . In the inner zone the counts were somewhat lower for both total organisms and Staph . aureus . Total counts on the floor from all areas of the inner zone were significantly higher just before the second operation than before the first operation on the same day . The total and Staph . aureus counts on walls, floors and lamps were the same after clean operations as after operations classified as "contaminated" or "dirty".

Jpn J Med Sci Biol, 1978 Feb, 31(1), 81 - 5
Enzyme-linked immunosorbent assay for detection of Clostridium botulinum toxin type A; Notermans S et al.; The enzyme linked immunosorbent assay using the so-called "double-sandwich technique" has been applied to determine botulinum toxin type A . By this assay, 50-100 mouse ip LD50 of toxin type A can be detected . No cross-reaction occurs with botulinum toxins of other types tested . In all probability this is due to the high specificity of the antiserum prepared against the toxic component of type A toxin.

Jpn J Med Sci Biol, 1978 Feb, 31(1), 1 - 15
Experimental botulism in chickens: the cecum as the site of production and absorption of botulinum toxin; Miyazaki S et al.; Highly purified preparations of Clostridium botulinum toxins were administered to chickens by various routes . Chickens were highly susceptible to type A toxin, but relatively resistant to toxins of other types . Type C toxin (12S) at a dose of 1 X 10(7) mouse ip LD50 failed to kill the chicken by the oral route . Oral administration of 10 or more of type A, C, or D spores killed normal chickens, whereas cecoligated chickens were insusceptible to oral administration of 10(6) spores . These results show that the site of production and absorption of botulinum toxin in chickens is the cecum . Peroral administration of spores of a type C strain cured of its prophages and producing the C2 factor only also killed normal chickens . Chickens appeared to the more susceptible to the C2 factor than to the C1 toxin . The C2 factor, therefore, may play more important role in chicken deaths from toxico-infection with type C organisms . The optimum temperature for growth of C . botulinum types C and D was found to be 40-42 C . Type C and D toxins were significantly more stable than type A toxin in the cecum contents with pH above 7 . These characteristics and the high density of distribution of type C spores in the environment may explain prevailing cases of type C botulism among broiler chickens.

Infect Immun, 1978 Feb, 19(2), 749 - 51
Acid precipitation of Clostridium botulinum type C and D toxins from whole culture by addition of ribonucleic acid as a precipitation aid; Iwasaki M et al.; The ratios of ribonucleic acid to protein contents of Clostridium botulinum type C, D, and E cultures were lower than those of type A, B, and F cultures . Addition of ribonucleic acid at 0.4 mg/ml to culture satisfactorily aided acid precipitation of type C and D toxins, but not that of type E toxin.

Zh Mikrobiol Epidemiol Immunobiol, 1978 Feb, (2), 86 - 90
{Protoplast fragmentation as one of the possible mechanisms of L-transformation of bacteria (Clostridium perfringens)}; Vysotskii VV et al.; As revealed, the action of lysozyme on the cells of Clostridium perfringens BP-6K led to the formation of not only typical spheroplasts, but also of cells whose peripheral parts of the cytoplasm were fragmented by membrane component . Small bodies framed by the membrane proper and containing granular and fibrillar components were formed . They were polymorphic in osmium treatment, and had smooth contours in preliminary use of aldehyde fixation . In the latter case a dense lumpy material analogous to the one which fills the periplasmic zone and serves as a rigid wall component formed at the surface of the protoplasm and bodies-fragments . In case of escape of the bodies into the external environment through the perforations in the cell wall the principal mass of the protoplast remains intact . The morphology of the bodies-fragments indicated a principal possibility of their autonomic existence . It is supposed that the phenomenon described could serve as one of the mechanisms of L-cell formation.

Appl Environ Microbiol, 1978 Feb, 35(2), 462 - 4
Observations on toxin and hemagglutinin produced by Clostridium botulinum type C; Oguma K et al.; In the culture fluid of a hemagglutinin-positive strain of Clostridium botulinum type C, two toxins of different molecular size, hemagglutinin positive and negative, were separated by sucrose density gradient centrifugation.

Appl Environ Microbiol, 1978 Feb, 35(2), 405 - 10
Clear, defined medium for the sporulation of Clostridium perfringens; Sacks LE et al.; A new, defined medium for the sporulation of Clostridium perfringens is presented . Sporulation levels exceeding 10(6) to 10(7) heat-resistant spores per ml were obtained for seven strains: PS49, PS52, FD-1, T-65, NCTC strains 8798, 8238, and 10240 . In the presence of theophylline, a methylxanthine, higher levels of heat-resistant spores were attained for strains PS49, PS52, FD-1, ant T-65; photomicrographs demonstrated a higher fraction of sporulating cells when these strains were grown in the presence of methylxanthines . Use of washed, highly diluted (less than 100 cells) inocula resulted in no reduction in spore yield . Strain KA3 grew well but sporulated poorly on this medium . The medium was clear and free of precipitate when small amounts (100 microgram/ml) of methylxanthine were incorporated.

Biochim Biophys Acta, 1978 Jan 18, 538(2), 212 - 25
Effect of hydrolytic enzymes and protein-modifying reagents on gonadotropin receptors in bovine corpus luteum cell membranes; Rao CV; Preincubation of membranes with various concentrations of pronase, trypsin, lipase, phospholipase A from Vipera russelli and from Crotalus durissus terrificus, phospholipase C from Bacillus cereus and from Clostridium welchii, acetic anhydride, 2,4-dinitrofluorobenzene and tetranitromethane resulted in a dose-dependent inhibition of 125I-labeled human choriogonadotropin binding . At the submaximal concentrations of enzymes and at both submaximal and maximal concentrations of protein-modifying reagents, the losses were always greater with 125I-labeled human choriogonadotropin than with 125I-labeled human lutropin . The inhibition of binding was a consequence of changes in the membranes rather than changes in the hormone caused by the agents being carried over to the final incubation . Inhibition of binding was non-competitive and irreversible . In untreated membranes, the 125I-labeled human choriogonadotropin binding was homogeneous (Kd = 1.7.10(-10) M; N = 60 fmol/mg protein) . Treatment of membranes with various enzymes and protein-modifying reagents except tetranitromethane resulted in heterogeneous binding . The number of available high affinity receptors was greatly reduced in every case . However, the affinity of these sites were either unchanged (trypsin, lipase, phospholipase A from V . russelli, dinitrofluorobenzene and the tetranitromethane) or decreased (pronase and acetic anhydride) . The newly appeared second receptor site had a Kd which varied from 3.2.10(-10) to 7.1.10(-9) M depending on the agent used, and the receptor numbers were low in all cases except acetic anhydride . Receptor occupancy conferred the receptors with marked protection against various hydrolytic enzymes, dinitrofluorobenzene and tetranitromethane . These data suggest that inhibition of binding by the above agents was primarily a consequence of changes in the receptor molecules themselves.

Arch Geschwulstforsch, 1978, 48(6), 538 - 51
{The determination of the oncolytic effect of clostridia with the hamster amelanotic Melanoma no.3--a quantitative study utilizing defined criteria of evaluation (author's transl)}; Fabricius EM et al.; With a view of satisfying the practical demands on a diagnostic tumour clostridium assay the development of a biological tumour test model is of prime importance . We are interested in a safe method for differentiating between oncolytic clostridia and clostridial strains exerting no oncolytic capacity . It was therefore attempted to develop a reproducible assay system using hamster A-Mel-3 as a reference tumour . This was finally attained by quantifying oncolysis taking into account the criterium of spontaneous onconecrosis of this tumour . The proposed oncolytic assay system was submitted to a mathematical regression analysis using different strains of clostridia as well as different batches of cultivated spores of one and the same strain . The results are critically compared with the current literature.

Acta Neurochir (Wien), 1978, 42(1-2), 123 - 5
The liquefaction (oncolysis) of malignant gliomas by a non pathogenic Clostridium; Heppner F et al.; Vascular glioblastomas become liquefied when contaminated with spores of the non-pathogenic Clostridium butyricum M 55 . The spores are administered by intracarotid injection . The oncolysis is complete one week after injection . The glioblastoma is converted into a brain abscess which is then operated on appropriately . Forty nine patients have been treated in this manner . The rate of recurrence, however, remained uninfluenced.

Zentralbl Bakteriol Naturwiss, 1978, 133(2), 125 - 34
Microbiological production of acetone-butanol by Clostridium acetobutylicum; Abou-Zeid AA et al.; Trials succeeded in raising the efficiencies of the fermentation medium, used in the fermentative production of acetone-butanol by Clostridium acetobutylicum . Egyptian black strap molasses (50.0% sugars) was suitable as carbon source in the fermentation medium, and (NH4)2SO4 was utilized with great success as inorganic nitrogen source . 140.0 g/l black strap molasses (about 7.0% sugars) and 3.0 g/l (NH4)2SO4 were the optimum concentrations for obtaining good yields of acetone and butanol . Molasses and (NH4)2SO4 were preferred because they are cheaper than the other carbon and organic nitrogen sources, used in the fermentative production of acetone-butanol . The percentage increase of the total solvents produced in the fermentation (production medium) was increased by 64.0 . The slop (by-product of the acetone-butanol fermentation after distillation) was re-used in the fermentation medium as organic nitrogen source and supported the microorganisms for a good production of acetone and butanol, while when stillage was used in the production medium, the total solvents output was less than that produced in the medium containing slop.

Arch Exp Veterinarmed, 1978, 32(1), 69 - 80
{Necrotizing enteritis in nursing piglets (Clostridium perfringens type C enterotoxemia) in industrialized swine breeding facilities}; Kohler B et al.; Outbreaks of necrotising enteritis, caused by Clostridium perfringens, Type C, among piglets on five industrialised breeding units are reported in this paper . Original diagnosis was confirmed by clinical, pathologico-anatomic, histological, and bacteriological tests as well as by the detection of beta toxin from the intestinal matter of piglets after death . Cl . per fringens, Type C, was successfully isolated from 71.9 per cent of all animals that had died of necrotising enteritis . Three forms were differentiated by pathologico-anatomic findings: 1 . haemorrhagic-necrotising enteritis with excessive bleeding of the intestinal wall and blood loss into the intestinal lumen; 2 . necrotising enteritis with small scattered foci of bleeding; 3 . necrotising enteritis without any bleeding . The first form usually developed concomitantly with peracute and acute disease in the first three days of age, whereas the third form was primarily recorded from piglets that had died, following subacute disease, between the fourth and ninth days after birth.

Nucleic Acids Res, 1978 Jan, 5(1), 221 - 39
HeLa DNA polymerase alpha activity in vitro: specific stimulation by a non-enzymic protein factor; Novak B et al.; A non-enzymic protein factor that increases the in vitro rate of synthesis by HeLa DNA polymerase alpha 15- to 30-fold with denatured DNA as template has been partially purified from the cytoplasmic fraction of HeLa cells . The stimulatory effect is highly specific for HeLa DNA polymerase alpha and for DNA templates that contain extensive regions of single-strandedness . Synthesis with denatured DNA as template presumably proceeds from 3'-hydroxyl termini formed at loop-back regions since the synthesized DNA product and template are covalently linked . The stimulatory protein factor chromatographs as a basic protein, has an approximate molecular weight of 30,000 daltons and binds with moderate affinity to denatured DNA cellulose, being eluted by o.4M NaCl . The purified factor lacks detectable DNA polymerase, exo- and endodeoxyribonuclease and RNA polymerase activities . It also does not promote helix-coil transitions with poly{d(A-T)} and Clostridium perfringens DNA.

Hoppe Seylers Z Physiol Chem, 1978 Jan, 359(1), 19 - 27
Properties of two Clostridia strains acting as catalysts for the preparative stereospecific hydrogenation of 2-enoic acids and 2-alken-1-ols with hydrogen gas; Bader J et al.; A Clostridium strain growing on crotonate/hydrogencarbonate, which is able to hydrogenate stereospecifically 2-enoates as well as other unsaturated compounds with hydrogen gas, has been isolated and methods for its propagation elucidated . For this strain and for Clostridium kluyveri DSM 555 grown on ethanol/acetate/hydrogencarbonate, samples of 200-l batches were assayed to determine the hydrogenation activity for (E)-2-methyl-2-butenoate and (E)-2-buten-1-ol as a function of time during the exponential and stationary growth phases . For the strain growing on crotonate/hydrogencarbonate, the hydrogenation rate as a function of substrate concentration, pH and temperature has been measured . Storage conditions for both strains are given.

Bol Med Hosp Infant Mex, 1978 Jan-Feb, 35(1), 151 - 8
{Finding of anaerobic bacteria in blood cultures}; Cob C et al.; Out of the blood cultures sent to the Bacteriology Laboratory of Hospital del Nino DIF, during the period of one year, in 1.4% of samples, isolation of anaerobic bacteria was obtained . Isolation was more frequent in infants under one year of age, but specially in newborns . The species most frequently found were Propionibacterium acnes, Bacteroides melaninogenicus and Clostridium subterminale . The presence of this finding in blood cultures of infants is noted, but there is no purpose to establish a clinical relationship with the finding of this group of bacteria.

J Bacteriol, 1978 Jan, 133(1), 26 - 32
Fermentation of fumarate and L-malate by Clostridium formicoaceticum; Dorn M et al.; The fermentation of fumarate and L-malate by Clostridium formicoaceticum was investigated . Growing and nongrowing cells degraded fumarate by dismutation to succinate, acetate, and CO2; on the other hand, only small amounts of succinate were detected when the organism was grown on L-malate . This dicarboxylic acid was mainly converted to acetate and CO2 . The fermentation balances were modified if bicarbonate or formate were present in the medium . When C . formicoaceticum was grown in the presence of both dicarboxylic acids, fumarate was consumed before L-malate . The latter was mainly converted to acetate, whereas fumarate was fermented to acetate and succinate . Molar growth yields were determined to be 6 g of dry weight per mol of fumarate and 8 g of dry weight per mol of L-malate fermented.

Enzyme, 1978, 23(1), 56 - 63
A purine N1-C6 hydrolase activity in the chicken egg yolk: a vestigial enzyme?
De Boeck S, Stockx J.
All attempts to prove the presence of xanthine oxidase and uricase in yolk preparations failed . We were able instead to show that yolk preparations could hydrolyze the N1-C6 bond of certain purine bases . In the case of xanthine, 4-ureido-imidazole-5-carboxylic acid and 4-ureido-imidazole are formed . Activity only becomes apparent during purification . An analogous enzyme was described earlier in Clostridium cyclindrosporum . The liver and the blood plasma of actively laying hens do not contain the enzyme . A scheme for the degradation of egg ribonucleic acids is presented.

Microbios, 1978, 23(91), 35 - 44
The flagellar protein of Clostridium tetani; Betts B et al.; Clostridium tetani ATCC 19406 was investigated with regard to the flagellar filaments produced by this anaerobic species . Flagellar filaments were removed from the cell bodies by hydrodynamic shear forces and purified by differential centrifugation . Exposure to acid was shown to result in disaggregation of the flagellar filaments into a preparation of flagellar protein containing 3.5% carbohydrate . The protein was judged homogeneous after examination by acrylamide gel electrophoresis in the presence of 4 M urea at several pH levels, and was shown to have a molecular weight of 35,000 daltons . Amino acid analyses indicated the absence of cysteine and tryptophan and a preponderance of acidic residues, epsilon-N-methyllysine was shown to be absent and the N-terminal amino acid was identified as alanine . Analysis of the C-terminal region indicated the sequence -Leu-Leu-Arg . These findings indicated that the obligate anaerobe C . tetani produced flagella filaments similar to previously studied filaments of aerobic and facultatively anaerobic bacteria.

Microbiol Immunol, 1978, 22(10), 591 - 6
Sporulation and C2 toxin production by Clostridium botulinum type C strains producing no C1 toxin; Nakamura S et al.; All of the 8 strains that were previously assumed to be nontoxigenic Clostridium botulinum type C were re-examined for their toxigenicity and were demonstrated by trypsinization of the culture filtrates to produce C2 toxin under improved cultural conditions . One per cent glucose added to trypticase peptone medium enhanced C2 toxin production . The larger the spore population, the higher the C2 toxicity and when spore population was smaller than a level of 10(4)/ml, no C2 toxicity was demonstrated . The C2 toxin was produced only during sporulation and not during vegetative growth.

Arch Exp Veterinarmed, 1978, 32(1), 155 - 70
{The quantitative composition of the gastrointestinal flora in piglets during the first weeks of life . 1 . The development of the gastrointestinal flora in naturally reared piglets}; Schulze F; Studies were conducted into the quantitative composition of the gastro-intestinal flora of 30 naturally raised piglets in four age groups . The gastro-intestinal tract was found to be "flooded" by Escherichia coli, Clostridium perfringens, micrococci, streptococci, proteus, and lactobacteria over the first hours after birth . Development of the intestinal flora was complete at the age of four days . The groups of germs recordable from naturally and artificially raised piglets were, basically, the same . Piglets, consequently, have no specific "lactoflora".

Appl Environ Microbiol, 1978 Jan, 35(1), 59 - 61
Enhancing nitrite inhibition of Clostridium botulinum with isoascorbate in perishable canned cured meat; Tompkin RB et al.; Addition of sodium isoascorbate to the formulation for perishable canned comminuted cured meat markedly enhanced the efficacy of nitrite against Clostridium botulinum . This effect was reproducible through a series of three tests . In one test it was found that the initial addition of 50 microgram of sodium nitrite per g plus isoascorbate was as effective as 156 microgram of sodium nitrite per g alone.

Arch Exp Veterinarmed, 1978, 32(6), 841 - 53
{Necrotizing enteritis in suckling pigs (Clostridium perfringens type C enterotoxemia) . II . Toxin formation, heat and drug resistance of Clostridium perfringens strains isolated from suckling pigs and broilers with necrotizing enteritis}; Kohler B; Nineteen Clostridium perfringens Type C strains and ten foreign control strains of subtypes C1, C3, and C4 were tested for their toxin formation and spore resistance to heat . The 19 Type C strains had been isolated from unweaned piglets in the context of necrotising enteritis outbreaks in the GDR . The Clostridium perfringens Type C strains formed beta-toxin, but they failed to form epsilon-toxin or gamma-toxin, alpha-toxin was successfully recorded from 15 of the 19 strains tested from unweaned piglets . The minor-lethal toxin fractions were also tested, with delta-toxin being recorded from all strains, non-alpha-delta-theta-toxin from six, theta-toxin from five, and K-toxin from one . Tests for delta-toxin, lambda-toxin, and mu-toxin were negative . The Clostridium perfringens Type C strains isolated in the GDR from unweaned piglets with necrotising enteritis were, basically, identical with those described in Denmark by von Hogh (1967) with regard to toxin formation . Clostridium perfringens strains cultured in broilers with necrotising enteritis were characterised by regular toxin production in the context of alpha, theta, delta as well as non-alpha-delta-theta . They failed to form beta, epsilon, gamma and lambda, while mu-toxin was formed by them quite irregularly . They, consequently, are Type A strains . Resistance to chloramphenicol and/or oxytetracycline was exhibited by 78.5 per cent of 237 tested Clostridium perfringens strains which had been isolated from unweaned piglets and broilers with necrotosing enteritis . Multiple resistance was recorded from 33.9 per cent . All strains were susceptible to penicillin, nitrofurantoin, and erythromycin.

J Hyg Epidemiol Microbiol Immunol, 1978, 22(2), 243 - 8
Production of toxic enzymes of Clostridium perfringens BP6K on technologically important media including a medium with Tween 80; Lettl A; The addition of 0.05--0.1% of Tween 80 to casein hydrolysate medium of Adams and Hendee raises and production of Clostridium perfringens BP6K lecithinase by approximately 50% . The use of the silicon antifoam Lukosan makes the medium particularly suitable for cultivation in large volumes . Higher levels of theta haemolysin and lecithinase are formed on the casein hydrolysate medium than on the meat medium . All factors except the theta and lambda components are farily stable during cultivation.

J Hyg Epidemiol Microbiol Immunol, 1978, 22(1), 63 - 72
Preparation of crude Clostridium histolyticum collagenase for therapeutic purposes; Lettl A; The production of a freeze-dried enzymatic preparation from the category of crude collagenases has been described . The method is based on the utilization of a highly proteolytic Clostridium histolyticum strain whose products have more advantageous properties for therapeutic purposes than the products of the strain commonly used as yet.

Scand J Infect Dis, 1978, 10(2), 119 - 25
The clinical importance of anaerobic bacteria in wound infections after gastrointestinal surgery; Schwan A et al.; Wound cultures from 54 patients with infections after gastrointestinal surgery were examined . Cultures from wounds after surgery on the upper gastrointestinal tract grew few organisms, mainly aerobic gram-positive cocci . Culture from wounds on the lower gastrointestinal tract grew strains of bacteria, aerobic and anaerobic gram-negative rods dominating . Indirect immunofluorescence studies on acute and convalescent phase sera showed significant immune response against Bacteroides fragilis in a majority of cases . Immune response against anaerobic cocci was seldom found . Very high antibody titres against Clostridium perfringens were often found, both in patients' and control sera.

J Clin Microbiol, 1978 Jan, 7(1), 23 - 7
Detection of alcohols and volatile fatty acids by head-space gas chromatography in identification of anaerobic bacteria; Larsson L et al.; A head-space gas chromatographic technique for the analysis of volatile bacterial metabolites is described . Bacteroides fragilis, Clostridium perfringens, and Propionibacterium acnes, cultured in a glucose-containing peptone yeast extract medium, were studied . The head-space technique was compared with the injection of the complete liquid culture medium, and solvent extracts thereof, into the gas chromatograph . Volatile fatty acids could be detected by all three methods, whereas alcohols produced by C . perfringens and P . acnes were detectable only in the head-space chromatograms . Both FFAP and Porapak Q were used as gas chromatography stationary phase . Porapak Q was found more suitable than FFAP for the separation of alcohols . The head-space technique requires a minimum of preparation before the analysis and is well suited to automation.

Acta Pharmacol Toxicol (Copenh), 1978 Jan, 42(1), 23 - 34
Inhibition of noradrenaline-stimulated lipolysis and cyclic AMP accumulation in isolated rat adipocytes by purified phospholipase C and theta-toxin from Clostridium perfringens; Freholm BB et al.; Purified phospholipae C (phosphatidylcholine cholinephosphohydrolase, EC 3.1.4.3) and theta-toxin from Clostridium perfringens both inhibited noradrenaline-stimulated lipolysis and cyclic AMP accumulation in isolated rat adipocytes in a dose-dependent manner . The action of phospholipase C was gradual in onset, while the effect of theta-toxin was almost immediate . Phospholipase C, but not theta-toxin, hydrolyzed membrane phospholipids and inhibited adenylate cyclase (EC 4.6.1.1) in a crude membrane fraction from fat cells . The inhibitory effects of phospholipase C were associated with morphological alterations detectable by electron microscopy, whereas effects of theta-toxin were observed at a time when no clearcut morphological alterations could be observed . It is concluded that the two purified principles from C . perfringens, which are both present in commercial preparations of phospholipase C, antagonize noradrenaline-stimulated cyclic AMP accumulation and lipolysis . Although their exact mechanisms of action have not been elucidated, phospholipase C and theta-toxin have different modes of attack.

Int J Gynaecol Obstet, 1978, 15(4), 322 - 4
Clostridial sepsis after abortion with PGF2alpha and intracervical laminaria tents--a case report; Green SL et al.; A case of clostridial endomyometritis and sepsis necessitating total abdominal hysterectomy which occurred 12 hours following abortion induced with intraamniotic administration of prostaglandin F2 alpha and laminaria tent insertion is discussed . Cultures from cervical, blood, and surgical specimens all yielded Clostridium perfringens . Intrauterine contamination with this microorganism most likely followed the insertion of laminaria tents through the cervical os, which was colonized with C . perfringens . Since C . perfringens may be present in the microflora of the lower female genital tract, great care must be taken to cleanse this area prior to intracervical laminaria tent insertion.

Lancet, 1977 Dec 24-31, 2(8052-8053), 1312 - 4
Pseudomembranous colitis: Presence of clostridial toxin; Larson HE et al.; A toxin was found in the faeces of nine out of nine patients with pseudomembranous colitis and two out of two with antibiotic-associated non-specific colitis . The toxin was neutralised by Clostridium sordellii antitoxin but not by a commercial mixture of C . welchii, oedematiens, and septicum antitoxins . The in-vitro and in-vivo properties of pseudomembranous-colitis toxin closely resemble those of the toxin of C . sordellii.

Arch Microbiol, 1977 Dec 15, 115(3), 277 - 84
Some properties of formate dehydrogenase, accumulation and incorporation of 185W-tungsten into proteins of Clostridium formicoaceticum; Leonhardt U et al.; Formate dehydrogenase of Clostridium formicoaceticum used only methyl and benzyl viologen, but not NAD as electron acceptor . The S0.5 values were 0.9 X 10(-4) M for formate and 5.8 X 10(-3) M for methyl viologen . Using potassium phosphate buffer a pH-optimum of 7.9 was observed . The initial velocity of the formate dehydrogenase activity reached a maximum at 70 degrees C, whereas the activity was stable only up to 50 degrees C . The level of formate dehydrogenase in C . formicoaceticum was increased to its maximum when 10(-6) M selenite and 10(-7) M tungstate were added to a synthetic medium . Addition of molybdate instead of tungstate did not increase the level of formate dehydrogenase . 185W-tungsten was concentrated about 100-fold by C . formicoaceticum; molybdate had no major effect on the uptake of tungsten . 185W-tungsten was found almost exclusively in the soluble fluid and was predominantly recovered after chromatography in a protein of about 88000 molecular weight . Occasionally a labelled protein of low molecular weight was observed . Again molybdate added even in high molar excess did not influence the labelling pattern . No radioactivity peak could be obtained at the elution peak of formate dehydrogenase activity . The extreme instability of formate dehydrogenase prevented further purification.

J Am Vet Med Assoc, 1977 Dec 1, 171(11), 1157 - 60
Nitrites, nitrosamines, and meat; Engel RE; Food additive regulations are frequently misunderstood and are often the basis for widely disparate views . The US Department of Agriculture (USDA) does not accept the concept that zero health risk with food additives is a humanly attainable goal; however, if the USDA is to accept that zero health risk is not attainable, then the department must propose, through regulations, a socially acceptable level of risk . Nitrite is an example . On one hand, nitrite is toxic at high levels and can combine with amines to form nitrosamines, which have been shown to cause cancer in animals . On the other hand, nitrite has been shown to provide the necessary protection, in many products, against Clostridium botulinum . The USDA, however, is not content to accept this position as a final one; they will continue to urge additional studies designed to further refine and decrease nitrite usage or replace it with a safe substitute . Given adequate information as to the relative risks involved in the use of nitrite or a substitute, society itself should be in a position to make the determination as to its willingness to accept potential risks in its daily life.

Infect Immun, 1977 Dec, 18(3), 761 - 6
Development of antitoxin with each of two complementary fragments of Clostridium botulinum type B derivative toxin; Kozaki S et al.; Two fragments with molecular weights of 111,000 (fragment I) and 59,000 (fragment II) were separated from each other by gel filtration of dithiothreitol and urea-treated, trypsinized derivative toxin (molecular weight, 170,000) of the proteolytic Okra strain of Clostridium botulinum type B on a column of Sephadex G-200 (superfine) with a buffer containing dithiothreitol and urea . Upon removal of dithiothreitol and urea by dialysis, the two fragments reassembled to reconstruct the derivative toxin molecule . Both fragments were immunogenic, and both anti-fragments neutralized type B toxin . The neutralizing activities of both anti-fragment I and anti-fragment II were, however, lower than that of the anti-derivative toxin, suggesting that the molecular integrity of derivative toxin is essential for sufficient production of the neutralizing antibody . The immunological difference found between type B toxin from a proteolytic strain and that from a nonproteolytic strain was ascribed to the antigenic difference of fragment I.

Biken J, 1977 Dec, 20(3-4), 105 - 15
Isolation and some properties of nontoxigenic derivatives of a strain of Clostridium tetani; Hara T et al.; Nontoxigenic derivatives of a toxigenic strain of Clostridium tetani were isolated gy treatment with acridine orange, N-methyl-N'-nitro-soguanidine, rifampicin or ultraviolet light . The frequency of appearance fo non-toxigenic derivatives on these treatments was 0.8 to 3.2 per cent . The nontoxigenic derivatives peoduced all the same extracellular antigenic and protein components as the toxigenic parent strain, except the toxin and materials cross-reacting with the toxin . The nontoxigenic strains, like the toxigenic parent strain, were lyzed by trratment with mitomycin C . Bacteriophage was detected in the lysates of all the nontoxigenic derivatives produced with mitomycin C, and this bacteriophage was morphologically indistinguishable from that obtained from the toxigenic parent strain . The genetic factor controlling tetanus toxin production is discussed.

Can J Microbiol, 1977 Dec, 23(12), 1706 - 13
The effect of rifampicin on the developmental phases of germinating spores of Clostridum sp., MSp+; Hawirko RZ et al.; The effect of rifampicin on the developmental phases of germinating spores of Clostridium botulinum, MSp+, has been studied . At sublethal concentrations of rifampicin (0.05 ng/ml) the time periods required for outgrowth and vegetative growth was significantly prolonged because of the inhibition of RNA and protein synthesis . However, rifampicin had essentially no effect on DNA synthesis or on subsequent spore formation . Chemical analyses showed that the amount of protein present in vegetative cells of the rifampicin-treated cultures was twice as great as in the untreated cultures but the total protein content of endospores was the same in both cases . It was revealed in ultrastructural studies of rifampicin (0.1 ng/ml) treated cultures, examined after 22 h, that septum formation and normal cell division of the emerging cell was blocked and a few cells showed constriction which produced one normal and one protoplast-like daughter cell.

Appl Environ Microbiol, 1977 Dec, 34(6), 823 - 31
Cryogenic gamma irradiation of prototype pork and chicken and antagonistic effect between Clostridium botulinum types A and B; Anellis A et al.; Inoculated, irradiated pork (2,300 cans) and chicken (2,000 cans) pack studies were performed to establish the 12D dose for these foods . Each can was inoculated with a mixture of 10(6) spores of each of 10 strains of Clostridium botulinum (five type A and five type B), or a total of 10(7) spores . The cans received a series of increasing doses of gamma rays (60Co) at -30 +/- 10 degrees C; they were incubated for 6 months at 30 +/- 2 degrees C and examined for swelling, toxicity, and recoverable botulinal cells . The highest rate of swelling for both foods occurred within the first week of incubation, and maximum swelling was observed within 4 to 5 weeks . The minimal experimental sterilizing dose (ESD) based on flat, nontoxic sterile cans was 3.0 less than ESD less than or equal to 3.2 Mrad for pork and 4.0 less than ESD less than or equal to 4.2 Mrad for chicken . An analysis of the partial spoilage data by extreme-value statistics indicated with 90% confidence that the rate of spore death in the two foods was not a normal distribution, but appeared to favor a shifted exponential function . Based on the latter distribution, and assuming one most resistant strain in the mixture of 10 used, the 12D dose computed to 4.37 Mrad, with a shoulder of 0.11 Mrad, for pork and to 4.27 Mrad, with a shoulder of 0.51 Mrad, for chicken . An assumption that there were two or more most resistant strains in the inoculum progressively lowered the 12D dose . There was an apparent antagonism between the irradiated type A and B viable strains in the two foods . Cans with type B cells and toxin predominated over cans with type A cells and toxin, but cans with a mixture of type A and B toxins predominated over cans with a mixture of Type A and B cells . At the highest sublethal doses, only type A cells survived in pork, but in chicken there was a least one type B strain that was at least as resistant as type A strains.

Am J Clin Pathol, 1977 Dec, 68(6), 794 - 6
Intraleukocytic spore formation and leukocytic vacuolization during Clostridium perfringens septicemia; Kuberski TT; The case of a patient with fulminant Clostridium perfringens septicemia and intravascular hemolysis is presented . The organisms in the blood stream were in sufficient quantity to be detected on a peripheral blood smear, and some were found to be within leukocytes . In vivo spore formation by C . perfringens is thought to be uncommon; however, in this patient probable spore formation by this organism was observed within peripheral blood leukocytes . The peripheral polymorphonuclear leukocytes from this patient were also markedly vacuolated.

J Biochem (Tokyo), 1977 Dec, 82(6), 1663 - 71
Studies on nitrate reductase of Clostridium perfringens . Purification, some properties, and effect of tungstate on its formation; Seki-Chiba S et al.; Nitrate reductase (NaR) linked to reduced methyl viologen from Clostridium perfringens was purified by ammonium sulfate precipitation . DEAE-cellulose chromatography, disc electrophoresis on polyacrylamide gel, and triple DEAE-Sephadex chromatography . The specific activity was increased 1,200-fold with a yield of 9% . The purified preparation was nearly homogeneous in disc electrophoresis . It was brown, and its spectrum showed a slight shoulder near 420 nm as well as a peak at 280 nm . The molecular weight was found to be 90,000 based on s020,w (5.8S) and 80,000 by Sephadex G-100 gel filtration . In SDS-polyacrylamide electrophoresis, it showed only a single band with a molecular weight of 90,000; it had no subunit structure . The isoelectric point was pH 5.5, and the optimum pH was 9 . Mn2+, Fe2+, Mg2+, and Ca2+ stimulated the activity . Km for nitrate was 0.10 mM, and nitrate was stoichiometrically reduced to nitrite in the presence of 2 mM Mn2+ . Ferredoxin fraction obtained from extracts of the bacterium was utilizable as an electron donor at pH 8 . Cyanide and azide inhibited the enzyme . The formation of NaR was induced by nitrate and inhibited by 0.5 mM tungstate, but recovered in the presence of 0.1 mM molybdate; NaR of C . perfringens appears to be a molybdo-iron-sulfur protein.

Biull Eksp Biol Med, 1977 Dec, 84(12), 695 - 8
{Immunogenicity and thymus-dependence of the polymerized alpha-toxoid of Clostridium perfringens}; Petrov RV et al.; Polymeric alpha-toxoid with a molecular weight of 450 000--600 000 was obtained by condensation of alpha-toxoid of Cl . perfringens, type A, with glutaric aldehyde . Experiments on guinea pigs showed that in the adsorbed preparations the immunogenic properties of both monomeric and polymeric alpha-toxoids are practically identical . The primary immune response after the immunization with nonadsorbed antigens was 3 times greater than that induced by the monomer . Polymerization of alpha-toxoid failed to change its thymus dependency.

Infect Immun, 1977 Dec, 18(3), 741 - 5
Purification of beta-toxin from Clostridium perfringens type C; Sakurai J et al.; Beta-toxin was purified about 340-fold from culture supernatant fluid of Clostridium perfringens type C with a yield of about 24% in terms of biologically active beta-toxin . The purification involved ammonium sulfate fractionation, gel filtration through Sephadex G-100, isoelectrofocusing in a pH 3 to 6 gradient, and immunoaffinity chromatography . The purified beta-toxin gave a single band on polyacrylamide gel electrophoresis.

Appl Environ Microbiol, 1977 Dec, 34(6), 843 - 8
Cultural and physiological characteristics of Clostridium botulinum type G and the susceptibility of certain animals to its toxin; Ciccarelli AS et al.; Strain 89 of Clostridium botulinum type G, isolated by Gimenez and Ciccarelli in 1969, was characterized culturally, biochemically, and toxigenically . It was motile, hemolytic asaccharolytic, weakly proteolytic, lipase and lecithinase negative, and it produced acetic, isobutyric, butyric, and isovaleric acids in peptone-yeast extract-glucose broth . No spores were seen in smears from solid or liquid media . Very low levels of toxin were produced in regular broth cultures, but dialysis cultures yielded 30,000 mouse 50% mean lethal doses (LD50 per kg, orally and subcutaneously, respectively; and for guinea pigs, 10,000 to 20,000 and 100 mouse LD50 per kig, intragastrically and intraperitoneally, respectively.

Lancet, 1977 Nov 26, 2(8048), 1103 - 6
Antibiotic-induced colitis implication of a toxin neutralised by Clostridium sordellii antitoxin; Rifkin GD et al.; A toxin(s) has been demonstrated in the stools of two patients with antibiotic-associated colitis . This toxin(s) was heat-labile, was rapidly lethal for hamsters, increased vascular permeability in rabbit skin, and was cytotoxic for cells in tissue-culture . It was neutralised by Clostridium sordellii antitoxin but not by antitoxins prepared against other clostridia; Escherichia coli, and Vibrio cholerae toxins . These characteristics were identical to those of a toxin implicated in the aetiology of antibiotic-induced colitis in the hamster . One patient improved rapidly after treatment with oral vancomycin, and at the same time the toxin disappeared from the stool.

Lancet, 1977 Nov 26, 2(8048), 1099 - 1102
Outbreak of necrotising enterocolitis caused by Clostridium butyricum; Howard FM et al.; 12 hospital-born babies had necrotising enterocolitis, of varying severity, within six weeks, 5 of them within ten days . The usually described predisposing causes were absent in most, though no baby was exclusively breast -fed . Evidence of the presence of Clostridium butyricum was found in the blood of 9 out of 10 babies examined . Cl . butyricum is probably a primary, not a secondary invader.

Z Rechtsmed, 1977 Nov 18, 80(3), 191 - 5
Immunochemical studies on blood group A substance from human hair; Kishi K et al.; Blood group A-active substance was extracted from urea-treated human hair uith methanol-ethyl ether 1:1, v/v) or chloroform-methanol (1:1, v/v) . The serological activity of blood group A substance in the hair was destroyed by A-decomposing enzyme from Clostridium tertium with concomitant development of blood group H activity . It is concluded therefore that the extract from the hair of group A contained blood group A-active glycolipid with N-acetylgalactosamine as the non-reducing sugar.

Biochim Biophys Acta, 1977 Nov 1, 470(3), 342 - 56
Asymmetry of the phospholipid bilayer of rat liver endoplasmic reticulum; Higgins JA et al.; The phospholipids of intact microsomal membranes were hydrolysed 50% by phospholipase C of Clostridium welchii, without loss of the secretory protein contents of the vesicle, which are therefore not permeable to the phospholipase . Phospholipids extracted from microsomes and dispersed by sonication were hydrolysed rapidly by phospholipase C-Cl . welchii with the exception of phosphatidylinositol . Assuming that only the phospholipids of the outside of the bilayer of the microsomal membrane are hydrolysed in intact vesicles, the composition of this leaflet was calculated as 84% phosphatidylcholine, 8% phosphatidylethanolamine, 9% sphingomyelin and 4% phosphatidylserine, and that of the inner leaflet 28% phosphatidylcholine, 37% phosphatidylethanolamine, 6% phosphatidylserine and 5% sphingomyelin . Microsomal vesicles were opened and their contents released in part by incubation with deoxycholate (0.098%) lysophosphatidycholine (0.005%) or treatment with the French pressure cell . Under these conditions, hydrolysis of the phospholipids by phospholipase C-Cl . welchii was increased and this was mainly due to increased hydrolysis of those phospholipids assigned to the inner leaflet of the bilayer, phosphatidylethanolamine and phosphatidylserine . Phospholipase A2 of bee venom and phospholipase C of Bacillus cereus caused rapid loss of vesicle contents and complete hydrolysis of the membrane phospholipids, with the exception of sphinogomyelin which is not hydrolysed by the former enzyme.

Postgrad Med, 1977 Nov, 62(5), 233 - 7
Tetanus in a 67-year-old man; Kinahan CC et al.; A fatal case of generalized tetanus in a 67-year-old man is presented . The incubation period following finger damage was short, and the diagnosis was confirmed by the isolation of Clostridium tetani . Subsequently, a partial surgical amputation of the infected finger during the clinically active stage of the disease did not remove all the infecting organisms . Although convincing evidence of any previous active immunization was lacking, no form of tetanus antitoxin was given at the time of injury . This case illustrates the potential dangers of unfamiliarity with tetanus owing to its low incidence in developed countries.

J Infect Dis, 1977 Nov, 136(5), 701 - 5
Clindamycin-associated colitis due to a toxin-producing species of Clostridium in hamsters; Bartlett JG et al.; Clindamycin-associated enterocolitis in hamsters was studied to detect and characterize a transmissible agent . It was found that the disease could be transferred by cecal contents and filtrates of cecal contents (pore size of filter, 0.02 micron) obtained from animals after administration of clindamycin . Subsequent work showed that enterocolitis could be produced with broth cultures of a species of Clostridium recovered from cecal contents of animals with clindamycin-induced disease . The cell-free supernatant of this strain also caused enterocolitis . Cecal contents from animals with clindamycin-induced disease incubated with gas gangrene antitoxin failed to cause intestinal lesions . These experiments indicate that clindamycin-associated colitis in hamsters is due to a clindamycin-resistant, toxin-producing strain of Clostridium.

Poult Sci, 1977 Nov, 56(6), 1909 - 13
Efficacy of lincomycin feed medication for the control of necrotic enteritis in broiler-type chickens; Maxey BW et al.; Necrotic enteritis was reproduced in two trials, conducted in a penned research-type broiler facility, by growing broiler-type chickens on litter obtained from a commercial poultry house which had experienced a chronic necrotic enteritis mortality problem . In each trial, various concentrations of lincomycin in feed were evaluated for effectiveness in controlling necrotic enteritis . Lincomycin was evaluated at concentrations of 2 to 100 g./ton in Trial 1 and at concentrations of 2 and 4 g./ton in Trial 2 . In each trial, non-lincomycin medicated control groups were also included . Each trial included six treatment groups each consisting of four 60-bird replicates . The coccidiostat used in all groups in Trial 1 and in four of the six treatment groups of Trial 2 was the same as had been used on the farm which the litter had been obtained . No other medications were used in any groups . Clinical coccidiosis due to Eimeria brunetti and E . maxima was prevalent in both trials . Birds receiving lincomycin at a concentration of 2 g./ton, or higher, showed a significant reduction in mortality from necrotic enteritis when compared to birds in coccidiostat-control pens not receiving lincomycin medication . Clostridium perfringens was isolated from the litter in all pens and from the livers of birds dying from necrotic enteritis.

J Biochem (Tokyo), 1977 Nov, 82(5), 1217 - 33
Phospholipase C from Clostridium novyi type A . II . Factors influencing the enzyme activity; Taguchi R et al.; The apparent activity of phospholipase C{EC 3.1.4.3} of Clostridium novyi type A toward phosphatidylcholine, sphingomyelin, and phosphatidylethanolamine increased in the presence of sodium deoxycholate (SDC) . The effects of divalent cations on phospholipase C activity were examined in detail at various concentrations of these cations . These effects varied with substrate . Hydrolysis of phosphatidylcholine by this enzyme significantly increased in the presence of Mg2+ or Ca2+ . Hydrolysis of sphingomyelin was inhibited by Ca2+, but increased in the presence of Mg2+ . Phosphatidylethanolamine-hydrolyzing activity increased only slightly in the presence of Mg2+ and Ca2+ . Zn2+ rather inhibited hydrolysis of these substrates . The effects of divalent cations and detergent appear to be directly related to the physical state of the phospholipid micelles used as substrates . When phosphatidylcholine, sphingomyelin, or phosphatidylethanolamine was used as a substrate, phospholipase C activity was completely inhibited by 2.5 mM EDTA or o-phenanthroline (concentration in the final incubation mixture: 0.5 mM), and was fully restored by Zn2+ alone . Both Ca2+ and Mg2+ were ineffective for reactivation . The isoelectric point of the enzyme was 7.1 +/- 0.1.

J Bacteriol, 1977 Nov, 132(2), 444 - 52
Levels of acetyl coenzyme A, reduced and oxidized coenzyme A, and coenzyme A in disulfide linkage to protein in dormant and germinated spores and growing and sporulating cells of Bacillus megaterium; Setlow B et al.; Dormant spores of Bacillus megaterium were found to contain approximately 850 pmol of coenzyme A (CoA) per milligram of dry weight . Of this total, less than 1.5% was acetyl-CoA, 25% was CoA-disulfide, 43% was in disulfide linkage to protein, and the remainder was the free thiol . Dormand spores of Bacillus cereus and Clostridium bifermentans contained 700 and 600 pmol of CoA per milligram of dry weight, respectively; in both species approximately 45% of the CoA 45% of the CoA was in disulfide linkage to protein . During germination of spores of all three species, greater than 75% of the CoA-protein disulfides were cleaved . In B . megaterium, cleavage of these disulfides during spore germination did not require exogenous metabolites and occurred at about the same time as the initiation of germination . Much of the CoA was converted to acetyl-CoA at this time . Dormant spores also contained reduced nicotinamide adenine dinucleotide-dependent CoA-disulfide reductase at levels higher than those in other stages of growth . The level of total CoA in the growing cells was two- to three-fold higher than in spores . This level remained constant throughout growth and sporulation, but less than 2% of the total cellular CoA was in disulfide linkage to protein until late in sporulation . The CoA-protein disulfides accumulated exclusively within the developing spore at about the time when dipicolinic acid was accumulated.

Appl Environ Microbiol, 1977 Nov, 34(5), 600 - 1
Comparative dose-survival curves of representative Clostridium botulinum type F spores with type A and B spores; Anellis A et al.; Radiation survival data of proteolytic (Walls 8G-F) and non-proteolytic (Eklund 83F) type F spores of Clostridium botulinum were compared with dose-response data of radiation-resistant type A (33A) and B (40B) spores . Strain Eklund 83F was as resistant as strain 33A, whereas strain Walls 8G-F was the most sensitive of the four strains tested . The methods suggested for computing both an initial shoulder and a D value for the dose-survival curves yielded results comparable to the graphic techniques used to obtain these two parameters.

Poult Sci, 1977 Nov, 56(6), 2052 - 5
Effect of freezing on incidence and levels of Clostridium perfringens in mechanically deboned chicken meat; Lillard HS; Paucity of information about the effect of plant practices on microbiological condition of comminuted chicken meat prompted this investigation . Commercially deboned chicken backs and necks were analyzed for levels of aerobic organisms, incidence and levels of Clostridium perfringens vegetative cells and spores before and after 4-6 weeks at -23 degrees C . Initially vegetative cells were isolated more frequently than spores . Frozen storage significantly reduced incidence and levels of vegetative cells and spores but did not affect levels of aerobic organisms . After frozen storage (a common industrial practice), with good food handling practices, C . perfringens should pose no undue hazard when comminuted chicken meat is incorporated into other food products.

Zentralbl Bakteriol {Orig A}, 1977 Nov, 239(3), 375 - 8
Media supplemented with nalidixic acid for the isolation of gram negative anaerobic bacteria; Nadaud M; The minimum inhibitory concentrations (MIC) of nalidixic acid were determined for thirty-four strains of Clostridium sp . by an agar dilution technique . The MIC range of Clostridium perfringens ran from 0.03 microgram/ml to 64 microgram/ml, according to a bimodal distribution . One half of the strains was inhibited by 0.25 microgram/ml . 64 microgram/ml was able to inhibit all the strains tested . The author suggests a systematic utilization of media with and without 40 microgram/ml nalidixic acid . These two media were parallely inoculated with clinical specimens where anaerobic bacteria were suspected, and allowed the isolation of gram negative anaerobes.

J Gen Microbiol, 1977 Nov, 103(1), 45 - 50
Increased numbers of heat-resistnat spores produced by two strains of Clostridium perfringens bearing temperate phage s9; Stewart AW et al.; Sporulation kinetics and spore heat resistance data were compared for a lysogenic strain of Clostridium perfringens, s9, before and after curing with ultraviolet irradiation . The cured strain showed the same growth rate in broth media as the lysogenic strain but took 6 h longer to form refractile spores . For lysogenized and cured strains the percentages of refractile spores produced that were heat-resistant (80 degrees C for 15 min) were 50 and 0.2, respectively . When reinfected with the temperature phage, the cured strain produced spores in 2 to 3 h, like the original lysogenic culture, and 10% of the spores produced were heat-resistnat.

J Biochem (Tokyo), 1977 Nov, 82(5), 1225 - 30
Hydrolytic action of phospholipases on bacterial membranes; Taguchi R et al.; Substrate specificities of phospholipases C{EC 3.1.4.3} from Clostridium novyi, Clostridium perfringens, Bacillus cereus, and Pseudomonas aureofaciens were studied under the same conditions . Phospholipases C from Clostridium novyi and Bacillus cereus show wide substrate specificities while those of Clostridium perfringens and Pseudomonas aureofaciens show relatively narrow specificities . On the basis of these results, the hydrolytic actions of these phospholipases on membrane lipids of Escherichia coli, Bacillus cereus, and Clostridium novyi were examined under the same conditions . The enzymes of Clostridium novyi and Bacillus cereus attacked all the membranes and their lipid extracts, hydrolyzing phosphatidylethanolamine, phosphatidylglycerol, lyso-phosphatidylethanolamine, and o-aminoacylphosphatidylglycerol . Phospholipase C from Pseudomonas aureofaciens attacked these three membranes and their lipid extracts, hydrolyzing phosphatidylethanolamine . Phospholipase C from Clostridium perfringens hardly attacked the phospholipids of these bacterial membranes . However, phospholipase C from Clostridium perfringens hydrolyzed phosphatidylethanolamine in a mixture containing lipid extract from Escherichia coli membrane and purified phosphatidylcholine from egg yolk.

J Clin Microbiol, 1977 Nov, 6(5), 494 - 8
Anaerobes in human biliary tracts; England DM et al.; During a 2-year period, 1,892 patients underwent biliary tract surgery at the Mayo Clinic . Both aerobic and anaerobic cultures of bile were performed in 371 of these patients . Sixty-nine percent of the cultures were positive, and 41% (117) of these grew anaerobes, although they were present in pure culture only twice . Mixed cultures most commonly contained four different organisms (three aerobes and one anaerobe) . Bacteroides fragilis was the single most commonly isolated anaerobe and ranked fourth in terms of overall isolates behind Escherichia coli, group D streptococci, and Klebsiella B . fragilis accounted for 7.0% of the total group D streptococci, and Klebsiella . B . fragilis accounted for 7.0% of the total aerobic and anaerobic isolates and was present in 21% of all positive cultures . Pseudomonas aeruginosa and Clostridium perfringens ranked fifth and sixth, providing 6.5 and 5.9% of all isolates, respectively . This study demonstrates the frequent presence of anaerobes in patients with bactibilia and suggests that they be considered in the formulation of antimicrobial therapy for infections involving human biliary tracts.

Infect Immun, 1977 Nov, 18(2), 549 - 51
Physical changes in the epsilon prototoxin molecule of Clostridium perfringens during enzymatic activation; Worthington RW et al.; Enzymatic activation of Clostridium perfringens epsilon prototoxin removed a small basic part of the molecule, causing a slight change in molecular weight (32,700 to 31,200) and a large change in isoelectric point (from pH 8.02 into fractions of 5.36 and 5.74).

J Infect Dis, 1977 Nov, 136(5), 661 - 6
Regional localization of activity of Clostridium perfringens type A enterotoxin in the rabbit ileum, jejunum, and duodenum; McDonel JL et al.; Rabbit ileal, jejunal, and duodenal loops were exposed to purified enterotoxin from Clostridium perfringens type A and then perfused for comparative analysis of effects of the enterotoxin on each region of the intestine . Ileal loops responded with enhanced net secretion of fluid and sodium, inhibition of chloride and glucose uptake, and substantial sloughing of epithelial cells . The jejunum responded with fluid secretion, enhancement of sodium secretion only during the first 20 min, inhibition of chloride and glucose uptake, and substantial sloughing of epithelial cells . In the duodenum, transport of fluid, sodium, and chloride was significantly altered only during the first 20 min of perfusion, and significant inhibition of glucose uptake varied from one period to another . Epithelial damage was much less than that seen in the jejunum or ileum . Levels of fluid protein in all three sections corresponded closely to extent of tissue damage . In general, it was found that the severity of response to fixed doses of enterotoxin varied as follows: ileum greater than jejunum greater than duodenum.

Biochemistry, 1977 Nov 1, 16(22), 4879 - 82
Effect of pressure upon the fluorescence of various flavodoxins; Visser AJ et al.; The effects of hydrostatic pressure in the range of 10(-3) to 11 kbar on the fluorescence of flavodoxins from Peptostreptococcus elsdenii, Desulfovibrio vulgaris, Azotobacter vinelandii, and Clostridium MP were investigated . The first three flavoproteins showed under high pressure enhancements of flavin fluorescence of over 50 times resulting from the release of flavin mononucleotide from the protein complex . The Clostridial flavodoxin showed a very much smaller fluorescence change . At pH 7.5 the high-pressure fluorescence changes of the flavodoxins of D . vulgaris and P . elsdenii were not reversed by decompression, but in A . Vinelandii the pressure changes were over 80% reversible . At pH 5 over 80% reversibility was restored to the flavodoxins of D . vulgaris and P . elsdenii, although the pressure dependence of the fluorescence changes was very similar in the reversible and irreversible cases . The midpoint pressures in the reversible reactions were 4.7 kbar (D . vulgaris), 8.7 kbar (P . elsdenii), and 10.6 kbar (A . vinelandii) indicating specific differences in the flavin binding regions . Apparent volume changes in these reactions were 65-75 mL/mol indicating participation of a large fraction of the protein in the pressure-induced changes . The irreversible changes are not related to protein aggregation and are believed to result from a pressure-dependent covalent modification, not yet characterized, of the flavin binding region of the protein.

Biochim Biophys Acta, 1977 Oct 26, 494(2), 301 - 13
Theta-toxin of Clostridium perfringens . I . Purification and some properties; Yamakawa Y et al.; Theta-Toxin, an oxygen-labile hemolysin produced by Clostridium perfringens, was purified 3300 fold from culture filtrate by successive chromatography on DEAE-Sephadex A-50 and Sephadex G-150 . The purified toxin gave two distinct bands in disc electrophoresis, while the same material, after mild reduction with dithiothreitol, yielded a single band, indicating that the purified theta-toxin contained, as well as a reduced, active form, an oxidized, inactive form of toxin . These two forms of the toxin had a similar, if not identical molecular size . The purified preparation gave a single band in a sodium dodecyl sulfate polyacrylamide gel electrophoresis and formed a single precipitin line with National Standard gas gangrene (C . perfringens) antitoxin . By sodium dodecyl sulfate polyacrylamide gel electrophoresis, the molecular weight of theta-toxin was estimated to be 51 000, the value being in exact accordance with that obtained by amino acid analysis . The amino acid composition of theta-toxin was very close to that of cereolysin, an oxygen-labile hemolysin produced by Bacillus cereus . The amino-terminal residue of theta-toxin was lysine as determined by the Dansyl method.

J Biol Chem, 1977 Oct 25, 252(20), 7089 - 92
The amino acid sequence of Clostridium pasteurianum iron protein, a component of nitrogenase . II . Cyanogen bromide peptides; Tanaka M et al.; A total of 10 cyanogen bromide peptides were isolated from the S-beta-carboxymethyl iron protein of nitrogenase . Purification of these peptides was performed mainly by gel filtration on Sephadex G-50; by ascending paper chromatography using the solvent system of pyridine, isoamyl alcohol, 0.1 M ammonium hydroxide; and also, in some cases, with additional steps such as anion exchange column chromatography on Dowex 1-X2 or ascending paper chromatography in an acidic solvent system or by pyridine precipitation of the cyanogen bromide fragment . Sequenator analyses of three large cyanogen bromide peptides (53 to 72 residues) provided tryptic peptide overlap data for the inner portion of the protein . The cyanogen bromide peptides accounted for all of the 273 amino acid residues which were present in the tryptic peptides isolated from carboxymethyl-iron protein (Tanaka, M., Haniu, M., Yasunobu, K . T., and Mortenson, L . E . (1977) J . Biol . Chem . 252, 7081-7088).

J Biol Chem, 1977 Oct 25, 252(20), 7081 - 8
The amino acid sequence of Clostridium pasteurianum iron protein, a component of nitrogenase . I . Tryptic peptides; Tanaka M et al.; A total of 25 tryptic peptides was isolated from the S-beta-carboxymethyl derivative of Clostridium pasteurianum iron protein (N2) . In order to obtain the various peptides in pure state, a combination of gel permeation, cation and anion exchange column chromatographic methods, as well as various ascending paper chromatographic methods were adopted . Sequence studies of the tryptic peptides were carried out mainly by a modified manual Edman degradation procedure and also by automated analysis, carboxypeptidase digestion, and by hydrazinolysis . Thus, 242 residues (88.6%) out of a total of 273 amino acid residues were sequenced in the present study . The sum of the amino acid residues in the tryptic peptides isolated from iron protein (N2) accounted for the 273 amino acid residues present in the iron protein.

JAMA, 1977 Oct 24, 238(17), 1829 - 32
Coproexamination for botulinal toxin and clostridium botulinum . A new procedure for laboratory diagnosis of botulism; Dowell VR Jr et al.; Stool or serum specimens or both from 318 persons pertaining to 165 botulism investigations over a three-year period were examined . Botulinal toxin was detected in stools of 19 of 56 patients and in sera of 20 of 60 patients with clinical botulism; it was not detected in specimens from 246 persons with an illness other than botulism or well contacts of patients . Clostridium botulinum was identified in stools of 36 of 60 clinical botulism patients and in four of 27 asymptomatic contacts of patients with botulism victims, but not in stools of 65 persons not associated with confirmed botulism . When stool and serum samples were examined, confirmatory evidence was obtained for 72.9% of the botulism cases . Detection of botulinal toxin or C botulinum in the stool of a persons should be considered evidence supporting the clinical diagnosis of botulism.

Am J Vet Res, 1977 Oct, 38(10), 1625 - 7
Experimental induction of bacillary hemoglobinuria in cattle; Erwin BG; An open-surgery technique for intrahepatic inoculation of Clostridium haemolyticum spores, suspended in calcium chloride as the hepatic debilitant, was used to produce bacillary hemoglobinuria in cattle . All calves (n=3) died of the disease, and the controls (n=2) given calcium chloride without spores survived . Clinical signs and gross pathologic changes produced by this method resembled those described for the disease in its natural form.

Arch Ophthalmol, 1977 Oct, 95(10), 1788 - 9
Ocular involvement in wound botulism; Miller NR et al.; A 7-year-old girl developed bilateral ptosis, total ophthalmoplegia, and fixed, dilated pupils associated with bulbar paralysis and generalized weakness six days after she sustained a compound supracondylar fracture of the right humerus . Nerve conduction studies showed a facilitated muscle action potential after repetitive nerve stimulation . Blood cultures were negative . Although the wound site appeared noninfected, the wound was explored . Clostridium botulinum, type B, grew from cultures taken from the depths of the wound . The patient recovered fully with supportive care, and EEG abnormalities present during the acute phase of the illness disappeared.

Appl Environ Microbiol, 1977 Oct, 34(4), 377 - 81
Recovery of spores of Clostridium botulinum in yeast extract agar and pork infusion agar after heat treatment; Odlaug TE et al.; Yeast extract agar, pork infusion agar, and modifications of these media were used to recover heated Clostridium botulinum spores . The D- and z-values were determined . Two type A strains and one type B strain of C . botulinum were studied . In all cases the D-values were largest when the spores were recovered in yeast extract agar, compared to the D-values for spores recovered in pork infusion agar . The z-values for strains 62A and A16037 were largest when the spores were recovered in pork infusion agar . The addition of sodium bicarbonate and sodium thioglycolate to pork infusion agar resulted in D-values for C . botulinum 62A spores similar to those for the same spores recovered in yeast extract agar . The results suggest that sodium bicarbonate and sodium thioglycolate should be added to recovery media for heated C . botulinum spores to obtain maximum plate counts.

JACEP, 1977 Oct, 6(10), 453 - 4
Infantile botulism; Fisher CJ Jr et al.; Botulism has not traditionally been considered as occurring in infants under one year of age because they generally do not ingest foods potentially containing preformed Clostridium botulinum toxin . We report a case of infantile botulism in a 3 1/2 month old infant who presented as a "floppy baby," and discuss the probable pathobiology involved.

Am J Vet Res, 1977 Oct, 38(10), 1515 - 7
Vaccination of cattle and sheep with a combined Clostridium perfringens types C and D toxoid; Kennedy KK et al.; Cattle and sheep (30 each) were vaccinated with a combined Clostridium perfringens type C and C perfringens type D toxoid . Vaccination and blood sample collections were made every 2 weeks over a period of 8 weeks . Increases in antitoxin titers occurred after the 2nd administration of the 2.0-ml combined product . Highest titers were recorded when the 2nd vaccinal dose was given 2 weeks after the first . This confirms reports that response to clostridial antigens is greater when the 2nd immunizing dose is delayed 2 to 6 weeks after the initial dose.

Zh Mikrobiol Epidemiol Immunobiol, 1977 Oct, (10), 37 - 43
{Biologic-economic study of processes for cultivating Clostridium perfringens . II . Comparison of the effectiveness of actual and predicted processes for cultivating microorganisms}; Zaporozhtsev LN et al.; The authors analyzed the efficacy of actual and prognosticated processes of Cl . perfringens, type A, cultivation: periodic, semicontinuous, continuous, and combined monocyclic . Comparative study permitted to choose the economically optimal processes, without any expenditure for their realization . Monocyclic combined process which could provide a great amount of active toxin at short periods proved to be the most advantageous among the cultivation processes studied.

Am J Dis Child, 1977 Oct, 131(10), 1104 - 6
Multiple hepatic tumors and peliosis hepatis in Fanconi's anemia treated with androgens; Shapiro P et al.; We report the case of a 13-year-old boy who was known to have Fanconi's anemia for five years . For treatment of this condition he was given androgens and corticosteroids . Two months before his death, severe varicella developed complicated by pneumonia, jaundice, and prolonged fever; all of which resolved during a five-week hospitalization . Three weeks later he died of Clostridium septicum sepsis caused by necrotizing enterocolitis . At autopsy he was found to have multiple hepatocellular neoplasms . A striking feature of the neoplasms was cholestasis . The liver also showed peliosis hepatis . The association of the use of certain androgenic steroids with hepatic neoplasms histologically resembling hepatocarcinomas, but characterized by lack of metastases and apparent reversibility, suggests the desirability of a new nomenclature for these hepatocellular lesions.

Arch Microbiol, 1977 Sep 28, 114(3), 219 - 24
Differentiation between Clostridium acidiurici and Clostridium cylindrosporum on the basis of specific metal requirements for formate dehydrogenase formation; Wagner R et al.; The formate dehydrogenases of Clostridium acidiurici and of C . cylindrosporum coupled the oxidation of formate with the reduction of viologen dyes . The basal activity level was about 0.85 mumoles/min X mg of protein for both species . The level of formate dehydrogenase of C . acidiurici increased 12-fold when 10(-7) M tungstate and selenite were present during growth . Molybdate exerted no effect . On the other hand, molybdate and selenite were required to increase the formate dehydrogenase of C . cylindrosporum, and tungstate exhibitedan antagonistic effect in this organism . Growth on hypoxanthine generally depended on the addition of bicarbonate . Supplementation with tungstate and selenite accellerated growth of C . acidiurici and increased again the level of formate dehydrogenase . The addition of both, molybdate and selenite was necessary to initiate growth of C . cyclindrosporum and to form an active formate dehydrogenase . The differences in the requirement for metal ion supplementation to form high levels of formate dehydrogenase and their involvement in hypoxanthine degradation can be used to differentiate between C . acidiurici and C . cylindrosporum.

J Chromatogr, 1977 Sep 21, 139(2), 283 - 90
Purification of alpha-L-fucosidase from various sources by affinity chromatography; Jain RS et al.; An affinity column for alpha-L-fucosidases was constructed by linking p-amino-phenyl 1-thio-alpha-L-fucopyranoside to Sepharose 4B through linkers of succinyl 3,3'-diamino-dipropylamine . Excellent purification of alpha-L-fucosidase from rat epididymis, Clostridium perfringens and Limulus polyphemus (horse shoecrab) could be effected inone step with good yield . An affinity column purification step can be introduced at any point in published purification procedures . The purified enzyme is essentially free of other glycosidases and proteolytic enzymes . The column material is stable and can be reused for at least two years.

Biochem J, 1977 Sep 15, 166(3), 495 - 9
Calcium ion-dependent diacylglycerol accumulation in erythrocytes is associated with microvesiculation but not with efflux of potassium ions; Allan D et al.; Erythrocytes from several different species were exposed to Ca2+ and the bivalent-cation ionophore A23187 . The lipid composition, morphology and K+ permeability of the treated cells were investigated . Erythrocytes from human, rat, guinea pig and rabbit (a) showed an increased concentration of 1,2-diacyl-sn-glycerol and enhanced labelling of phosphatidate with 32P, (b) underwent echinocytosis and outward vesiculation, and (c) rapidly released much of their intracellular K+ . Pig cells showed only the K+ loss, and ox and sheep (high-K+) cells showed none of these Ca2+-evoked effects . All of the cells underwent stomatocytosis and inward vesiculation when treated externally with Clostridium perfringens phospholipase C . These results support the idea that there is a correlation between the asymmetric insertion of diacylglycerol (or ceramide) into the membrane and the shape-changes leading to microvesiculation, but they indicate that Ca2+-triggered K+ efflux and diacylglycerol production are unrelated events . Erythrocytes of chicken and turkey showed no Ca2+-stimulated K+ efflux . They showed slight ionophore A23187-stimulated vesiculation, but this appeared to be associated with the appearance in the membrane of ceramide rather than of diacylglycerol . Phospholipase C treatment caused very similar changes in morphology and phosphatidate labelling to those seen in mammalian erythrocytes.

Schweiz Med Wochenschr, 1977 Sep 3, 107(35), 1209 - 24
{Clostridium infections with and without manifest gas gangrene . Report on 77 infections in 76 patients}; Sonnabend O et al.; Systematic microbiological research and correlation of the histopathological findings obtained from random autopsies revealed 23 hitherto undetected clostridial infections including 11 cases of gas gangrene, 4 of septicemia, 3 of bacteremia, and 5 other clostridial infections . The knowledge gained from this study led to clinical diagnosis of several cases of gas gangrene which were confirmed bacteriologically and histologically . Of 8 hospital patients who were thus diagnosed in this surgical clinic, 7 recovered, including a case of gas gangrene of the abdominal wall . The problem in gas gangrene is timely clinical diagnosis . Little is known about gas edema illnesses which are not traumatically conditioned . Recognition of the local and general symptoms (local, violent, yet inappropriate pain in the wound, "unexplained" postoperative secondary bleeding, appearance of tachycardia wholly unrelated to the patient's temperature, sudden shock, rapid deterioration of patient's general condition, jaundice and rise in CPK) makes it possible to diagnose postoperative gas edema in time . 77 infections with isolation of clostridia, seen in 76 patients, are reported . On the basis of clinical and histopathological criteria they have been classified as follows: 22 cases with gas gangrene (clostridial myonecrosis), 16 cases with anaerobic cellulitis, 20 wound infections, 8 cases of septicemia, 5 of bacteriemia, 1 of tetanus, and 5 other clostridial infections.

Can Med Assoc J, 1977 Sep 3, 117(5), 483 - 9
Type A and type B botulism in the North: first reported cases due to toxin other than type E in Alaskan Inuit; Barrett DH et al.; Botulism outbreaks shown to be due to type A and type B toxin occurred in Alaska, a region previously known for only type E botulism . The outbreak due to type A toxin involved three people, two of whom died . The outbreak due to type B toxin involved nine people, none of whom died . Both outbreaks were in Inuit villages, and native foods were incriminated . The occurrence of these outbreaks strongly suggests that Clostridium botulinum, types A and B are indigenous to Alaska . The outbreaks underscore the need for initial treatment of patients with antitoxin that is trivalent (ABE), even in Arctic regions.

J Am Vet Med Assoc, 1977 Sep 1, 171(5), 431 - 2
Suspected infectious necrotic hepatitis (black disease) in Oregon cattle; Kelch WJ et al.; Suspected infectious necrotic hepatitis (black disease) in a herd of 436 cattle in Douglas County, Oregon, resulted in 79 deaths during a 2-week period . Although Clostridium novyi could not be isolated from hepatic lesions, the clinical course of the disease, gross and histopathologic findings, and fluorescent antibody identification of C novyi in various tissues were suggestive of the disease . The epizootic was preceded by a long drought, during which grazing conditions were sparse . A few days before the 1st dead animal was found, the drought was relieved by about 10 cm (4 in) of rainfall, resulting in the growth of young succulent grass . The cattle, attempting to eat this new grass lying close to the ground, consumed large quantities of soil . It was speculated that the soil contained C novyi and that the proliferation of these ingested organisms in necrotic tissue cuased by Fasciola hepatica resulted in fatal toxemia.

Biophys J, 1977 Sep, 19(3), 253 - 64
The iron-sulfur environment in rubredoxin; Bunker B et al.; The atomic environment around the iron site in the nonheme iron sulfur protein rubredoxin was studied by the extended X-ray absorption fine structure (EXAFS) technique . Within experimental error, the Fe-S bonds in oxidized Clostridium pasteurianum rubredoxin are the same as in the analogue anion {Fe(S2-o-xyl)2}-synthesized by Holm . The average Fe-S bond length is 2.267 +/- 0.003A and the root mean square deviation about this average due to structural disorder is 0.032 + 0.013 - 0.032.

Am J Med Sci, 1977 Sep-Oct, 274(2), 211 - 2
Pneumonia and empyema caused by Clostridium sordellii; File TM Jr et al.; A case of pleuropulmonary infection caused by Clostridium sordellii is reported for the first time . The clinical presentation with acute onset resembling pulmonary infarction, the absence of toxicity, hemolysis and shock, and response to penicillin and drainage was similar to that of patients with pleuropulmonary infection caused by C . perfringens.

Nord Vet Med, 1977 Sep, 29(9), 386 - 91
The occurrence of Clostridium botulinum type E in Finnish trout farms and the prevention of toxin formation in fresh-salted vacuum-packed trout fillets; Ala-Huikku K et al.; The occurrence of C . botulinum on two Finnish rainbow trout farms were studied . C . botulinum type E toxin was detected from samples of fish intestines in 10% and 4% of the samples and in 0% and 100% of dam bottom sediments, respectively . The toxin formation of inoculated C . botulinum type E in three different brands of commercial fresh-salted vacuum-packed trout fillets was also investigated . In the brand with a salt concentration of 2.7% (a w = 0.96) and no nitrate, the toxin was formed in two weeks at 10 and 20 degrees C . If the pH of the product was lowed to 4.90 and NaCl concentration increased to 5.67% (a w = 0.94%) no toxin was formed even without the use of nitrate . By adding 0.1% of sodium nitrate to the curing solution (0.046% nitrate in the product) the toxin formation was brought to an end . According to the results of the study C . botulinum type E presents a potential health risk for man as well as for fish also in Finland . Trout products must be manufactured and stored in such a way that the possibility of toxin formation is eliminated.

Can J Microbiol, 1977 Sep, 23(9), 1257 - 60
Inhibition of Clostridium botulinum types A and B hemagglutinins by sugars; DasGupta BR et al.; Chromatographically isolated hemagglutinins of Clostridium botulinum types A and B are serologically related but not identical . Of the sugars (5, 6, 12, 18 carbons, some derivatives, L and D forms) tested, only D-galactose and some of tis derivatives were inhibitors of these hemagglutinins . O-Nitrophenyl-beta-D-galactopyranoside and isopropyl-beta-D-thiogalactoside were the most potent inhibitors . The two hemagglutinins were bound tightly by p-aminophenyl-beta-D-thiogalactopyranoside coupled to CH-Sepharose 4B . The ligands to which these hemagglutinins bind were determined as the sugars which inhibited the hemagglutinating activity.

Appl Environ Microbiol, 1977 Sep, 34(3), 280 - 4
Thermal inactivation of ileal loop-reactive Clostridium perfringens type A strains in phosphate buffer and beef gravy; Bradshaw JG et al.; The thermal resistance of spore crops produced from each of two ileal loop-reactive strains of Clostridium perfringens type A was determined in two suspending vehicles consisting of 0.067 M (pH 7.0) phosphate buffer and a commercial beef gravy . D115.6 values obtained in buffer and enumerated after pretreatment with sodium ethylenediaminetetraacetate and recovery in plating medium containing lysozyme were two- to threefold greater than those obtained without this treatment . D115.6 values obtained with beef gravy were less than those obtained in buffer with or without lysozyme; however, the D98.9 and D104.4 values were 1.3 to 2 times greater than those obtained in buffer with lysozyme . The z values were within the ranges reported by previous investigators.

Immunology, 1977 Sep, 33(3), 407 - 12
Action of sphingomyelinase C and other lipid-specific agents as inhibitors of Fc binding and locomotion in human leucocytes; Wilkinson PC; The binding of antibody-coated chicken erythrocytes (EA) to human blood lymphocytes and monocytes is inhibited by pretreatment of the leucocytes with sphingomyelinase C . Inhibition of rosetting of neuraminidase-treated EA with neutrophils is also seen with this enzyme . The cholesterol-binding theta-toxin of Clostridium perfringens and pronase also inhibit EA-rosette formation, but less strongly than sphingomyelinase . The lipid-specific agents also inhibit chemotactic migration of leucocytes to casein and denatured HSA, whereas proteases and glycosidases do not . These results suggest that membrane lipids are important constituents of the binding sites for Fc fragments and for certain chemotactic factors and point to an important role for sphingomyelin in this binding.

Rev Can Biol, 1977 Sep, 36(3), 205 - 15
Properties of four temperate bacteriophages active on Clostridium perfringens type A; Paquette G et al.; Four temperature bacteriophages (designated as PF1, PF2, PF3 and PF4) were isolated from lysogenic strains of Clostridium perfringens type A . On the basis of plaque morphology, pH stability, DNase and RNase resistance, buoyant density, one-step growth parameters and electron microscope phage dimensions, it seems that these phages are different and unrelated . Calcium was required for better phage replication . Bacterial strain S107 appears to be the only UV-inducible strain as compared with the other three lysogenic strains . PF2 has a unique pattern of pH stability showing two optima values: one at pH 5 and the second at pH 8-9 . Generally, all four phages have a better resistance in acid than in alkaline pH values . The CsC1 equilibrium centrifugation patterns reveal low figures for phage PF1, PF2 and PF3 and show off the fact that PF4 lysates contain two viral particules different with respect to their densities, a property which other determinations failed to demonstrate . Each phage, except PF4, is well characterized by the parameters of the one-step growth cycle.

Minerva Med, 1977 Aug 4, 68(37), 2575 - 99
{Classification of acute bacterial enteropathies}; Altucci P et al.; The first part considers pathogenic microorganisms (Vibrio cholerae and parahaemolytic vibrio, Clostridium welchii, enteropathogenic E . coli, Shigella, Salmonella, other enterobacteria and pseudomonas . Yersinia, simply enterotoxic Staphylococcus and that producing acute enteritis) and the process of infection (formation of a surface link without endocellular penetration with elaboration of hexotoxins, formation of a surface link with subsequent intracellular penetration, submucosa penetration) . The second part discusses Salmonellae on the basis of personal experience . Particular attention is paid to current aspects of Salmonella microbiological pathomorphosis, the various isolated serotypes in relation to carriers or patients, biochemical atypias of Salmonellae strains, present-day aspects of resistance to chemoantibiotic treatment and the transfer of Salmonella Wien resistances.

Infect Immun, 1977 Aug, 17(2), 395 - 401
Clostridium botulinum type D toxin: purification, molecular structure, and some immunological properties; Miyazaki S et al.; Clostridium botulinum type D progenitor toxin was purified . The addition of ribonucleic acid to the whole culture helped initial acid precipitation of the toxin . As with type B, both L (16S) and M toxins (12S) obtained from a hemagglutinin-positive strain, whereas M toxin only was produced by a hemagglutinin-negative strain . M toxin (molecular weight, 300,000) consisted of one molecule each of a toxic (molecular weight, 170,000) and a nontoxic component (molecular weight, 130,000); L toxin consisted of both components plus hemagglutinin . The specific toxicity of M toxin was 5 X 10(8) mean lethal doses per mg of N; that of L toxin was 2.4 X 10(8) mean lethal doses per mg of N . These toxins were fully or nearly fully active, but in un-nicked form . Trypsinization caused nicking in the toxic component, forming a molecule made up of two peptide chains with molecular weights of 110,000 and 60,000; there was little or no increase in toxicity . The toxic component of type D was not antigenically related to that of type C, whereas the nontoxic component was antigenically indistinguishable from that of type C . The toxicities of both L nad M toxins of the hemagglutinin-positive strain were increased twofold by trypsinization . Neither toxin contained the C2 toxic factor elaborated by C and D strain.

Can J Microbiol, 1977 Aug, 23(8), 947 - 53
The intracellular reserve polysaccharide of Clostridium pasteurianum; Darvill AG et al.; An amylopectinlike polysaccharide (granulose) was the only glucan produced in significant quantities by six wild-type strains of Clostridium pasteurianum grown in glucose minimal medium . The intracellular polysaccharide granules laid down before sporulation contained only this amylopectin . No intracellular dextran was discovered in these wild-type strains, nor in a granulose-negative mutant strain of C . pasteurianum possessing an ADP glucose pyrophosphorylase (EC2.7.7.27) but lacking a granulose synthase (i.e . ADP glucose-alpha-1,4-glucan glucosyl transferase, EC2.4.1.21) . Furthermore, methylation analysis demonstrated that (1 leads to 6) linked alpha-D-glucose units accounted for less than 2% of the entire glucose content of these organisms.

Am J Dis Child, 1977 Aug, 131(8), 857 - 9
Botulism in infancy . Report of a case; McKee KT Jr et al.; A 22-day-old infant developed infant botulism characterized by profound weakness, hypotonia, respiratory arrest, areflexia, ptosis, pupils that responded poorly to light, and absent gag reflex . Stool examination yielded Clostridium botulinum type B organisms and type B toxin . Electromyography provided rapid diagnostic assistance . With supportive care, reovery was complete . This "new" disease probably is more common than now appreciated.

Infect Immun, 1977 Aug, 17(2), 415 - 24
Antagonistic effect of extremely oxygen-sensitive clostridia from the microflora of conventional mice and of Escherichia coli against Shigella flexneri in the digestive tract of gnotobiotic mice; Ducluzeau R et al.; Two extremely oxygen-sensitive strains of Clostridium sp., designated Clostridium E and P, were obtained from digestive microflora of conventional mice and found to constitute a barrier against Shigella flexneri SF-2 when associated in vivo with Escherichia coli K-12 . These and other simplified fractions of the conventional microflora were demonstrated to have an effect comparable to that of the total flora . When K-12 and Clostridium E were established in gnotobiotic mice before the introduction of SF-2, the latter was reduced to a level below detection in the digestive tract . Whe SF-2 was established first, the antagonistic effect exerted by Clostridium E and K-12 was variable and, apparently, related to the rate of establishment of Clostridium E . Mutants of SF-2 resistant to the barrier effect of Clostridium E and K-12 appeared at the end of 3 months when SF-2 was established in gnotobiotic mice alone or with K-12, and after only a week when SF-2 was associated only with Clostridium E . These results suggest that the bacterial antagonsim in this model is related to the production in vivo of an antibiotic substance active against SF-2 . It appears that the substance may be produced by Clostridium E, stimulated by K-12.

Cancer, 1977 Aug, 40(2), 950 - 3
Clostridium septicum infection in childhood leukemia: report of a case and review of the literature; Lehman TJ et al.; Overwhelming Clostridium septicum infection is a rare occurrence in children . It is seen almost exclusively as a complication of acute leukemia . A high index of suspicion in the leukemic child with an acute abdomen is the key to early diagnosis and improved survival . A case in a 13-year-old girl with acute myelogenous leukemia is reported and six pediatric cases in the literature were reviewed.

Appl Environ Microbiol, 1977 Aug, 34(2), 189 - 93
Increased spore yields of Clostridium perfringens in the presence of methylxanthines; Sacks LE et al.; The methylxanthines caffeine, theophylline, and isobutylmethylxanthine greatly increased spore yields of Clostridium perfringens strains FD-1, PS52, and PS49 when grown on Duncan-Strong medium or on a new casein-digest medium . Four other strains (KA3, and National Collection of Type Cultures strains 8798, 8238, and 10240) failed to show any significant increase when tested under similar conditions . The degree of sporulation increase was influenced by the carbohydrate energy source in some strains but not in others . Strain PS52 showed a large increase in spore yield when dextrin was the energy source but only a slight increase when raffinose served as the energy source . Strain FD-1 showed similar increases in spore yield with either dextrin or raffinose.

Appl Environ Microbiol, 1977 Aug, 34(2), 125 - 8
Rapid detection and quantitation of Clostridium perfringens enterostoxin by counterimmunoelectrophoresis; Naik HS et al.; Conditions for detection and quantitation of Clostridium perfringens enterotoxin by counterimmunoelectrophoresis are described . As little as 0.2 microgram of enterotoxin per ml could be detected . The test was found to be rapid, sensitive, specific and easy for the detection and quantitation of enterotoxin.

Mutat Res, 1977 Aug, 46(4), 261 - 4
Activation of a procarcinogen to a mutagen by cell-free extracts of anaerobic bacteria; McCoy EC et al.; The Salmonella mutagenicity assay can be coupled to cell-free preparations derived from anaerobic bacteria (Clostridium perfringens and Bacteroides fragilis) to activate a procarcinogen to a mutagen . This activity is destroyed by heating and by digestion with pronase and it is sensitive to oxygen . These findings indicate that the Salmonella mutagenicity assay can be adapted to the study of the role of anaerobes in the activation of carcinogens.

J Reprod Med, 1977 Aug, 19(2), 64 - 6
Fulminant puerperal sepsis associated with aplastic anemia: the case for prophylactic antibiotic therapy; Gardner P et al.; Fulminant puerperal sepsis due to Clostridium perfringens occurred in a primiparous 19-year-old woman who developed aplastic anemia during pregnancy . Although the risk of infectious complications among pregnant women with compromised host defenses has not been accurately determined, it appears to be increased, particularly in patients with granulocytopenia . It therefore seems reasonable to give antibiotic prophylaxis during the intrapartum period to immunosuppressed women who come to term . Consideration of the bacterial pathogens most likely to cause acute endometritis in the early puerperium has led us to recommend a short course (72 hours) of penicillin G, 4 million units q4h, and gentamicin, 1,5 mg/kg q8h, as an appropriate regimen for such patients.

Infect Immun, 1977 Aug, 17(2), 425 - 9
Preparative polyacrylamide gel electrophoresis purification of Clostridium perfringens enterotoxin; Enders GL Jr et al.; Preparative polyacrylamide gel electrophoresis has been used to purify the enterotoxin of Clostridium perfringens from Sephadex G-100 extracts . Purified toxin of high specific activity was eluted in 1 to 3 h, depending upon the length of the acrylamide gel used . Recovery of biological activity with this technique ranged from 80 to 90% . The purity and physical characteristics of the toxin were similar to those previously reported for the protein purified by other methods . Use of preparative electrophoresis will enable the production of larger amounts of high-specific-activity toxin in a shorter time than other currently available procedures . This method was also used to isolate a form of enterotoxin that has a mobility, relative to bromophenol blue tracking dye, of 0.87 to 0.90 in 7% acrylamide gels.

J Bacteriol, 1977 Aug, 131(2), 713 - 5
Spore coat protein and enterotoxin synthesis in Clostridium perfringens; Labbe RG et al.; Polyacrylamide gel profiles of Clostridium perfringens spore coat protein revealed four and occasionally five components . Pulse-chase experiments indicated that synthesis of coat protein polypeptide and enterotoxin was an early sporulation event . However, maximum synthesis occurred coincident with the onset of heat resistance.

J Bacteriol, 1977 Aug, 131(2), 693 - 5
Regulation of the arginine dihydrolase pathway in Clostridium sporogenes; Venugopal V et al.; Arginine deiminase activity was induced during the vegetative growth of Clostridium sporogenes . The enzyme was sensitive to catabolite repression . The other enzymes of the arginine dihydrolase pathway, namely, ornithine carbamoyl-transferase and carbamate kinase, did not show such variation.

J Bacteriol, 1977 Aug, 131(2), 463 - 72
Partial purification of ferredoxin from Ruminococcus albus and its role in pyruvate metabolism and reduction of nicotinamide adenine dinucleotide by H2; Glass TL et al.; Extracts of Ruminococcus albus were not able to convert pyruvate to acetyl phosphate, CO2, and H2 after passage through a diethylaminoethyl (DEAE)-cellulose column . Activity was restored by a brown protein fraction eluted from the column with 0.4 M Cl- . The protein was partially purified and shown to have the spectral and biological characteristics of ferredoxin . R . albus ferredoxin, Clostridium pasteurianum ferredoxin, and methyl viologen restored activity for pyruvate decomposition by DEAE-cellulose-treated R . albus extracts . R . albus or C . pasteurianum ferredoxin restored the ability of DEAE-cellulose-treated C . pasteurianum extracts to form H2 and acetyl phosphate from pyruvate . Ferredoxin-free extracts of R . albus reduced nicotinamide adenine dinucleotide (NAD) when supplemented with R . albus or C . pasteurianum ferredoxin or with methyl viologen . These extracts reduced NADP with H2 poorly unless both ferredoxin and NAD were added, which indicates the presence of an NADH:NADP transhydrogenase . Flavin mononucleotide and flavin adenine dinucleotide were rapidly reduced by H2 by ferredoxin-free extracts in the absence of ferredoxin.

Biochim Biophys Acta, 1977 Aug 1, 468(3), 502 - 6
Decreased iodination of the red cell surface following phospholipase C treatment; Reichstein E et al.; Human red blood cells were treated with phospholipase C from Clostridium welchii . Lipase concentrations which produced less than 1% hemolysis and 10-15% hydrolysis of the membrane phospholipids reduced markedly (greater than 80%) the accessibility of membrane proteins to the external surface as measured by lactoperoxidase-catalyzed iodination.

Arch Surg, 1977 Aug, 112(8), 965 - 7
Bacteriology of the human biliary tract and the duodenum; Lou MA et al.; Using the modern anaerobic transport media and meticulous culture techniques, 74 patients undergoing biliary tract surgery were studied . The biliary system was found to be sterile in 58 patients (78%) . Fifteen patients had 35 isolates of aerobic and facultative bacteria . The most common ones were Klebsiella, Enterococcus, and Escherichia coli . The only anaerobe isolated was Clostridium perfringens . Eight of 17 patients (47%) with acute cholecystitis and five of 49 patients (10%) with chronic cholecystitis, harbored bacteria in the biliary system . This study suggests that anaerobes are rare in the human biliary system; therefore, if antibiotic therapy is considered, aerobic coverage should suffice.

Appl Environ Microbiol, 1977 Aug, 34(2), 222 - 4
Degradation of lindane by cell-free preparations of Clostridium sphenoides; Heritage AD et al.; Cell-free preparation of Clostridium sphenoides degraded the insecticide lindane, the gamma-isomer of 1,2,3,4,5,6-hexachlorocyclohexane, to the gamma-isomer of 3,4,5,6-tetrachloro-1-cyclohexene . The activity appeared to be associated with the membrane fraction and required reduced glutathione . The tetrachlorocy-clohexene intermediate was further metabolized by the membrane fraction to unknown substances.

Jpn J Med Sci Biol, 1977 Aug, 30(4), 179 - 90
Response of type B and E Botulinum toxins to purified sulfhydryl-dependent protease produced by Clostridium botulinum type F; Ohishi I et al.; A sulfhydryl-dependent protease (SHP) was purified from a culture of Clostridium botulinum type F . The enzyme can activate type E progenitor toxin completely but type B progenitor toxin only partially . This may suggest that SHP by itself could completely activate the toxin of proteolytic C . botulinum types A and F in culture . The toxicity of type E progenitor toxin potentiated by the treatment with SHP persisted, whereas that of derivative toxin decreased rapidly by further incubation with SHP . This may indicate that only the progenitor toxin, the complex of the toxic and nontoxic components, activated by SHP withstands the subsequent exposure to the enzyme in cultures of proteolytic C . botulinum.

Infect Immun, 1977 Aug, 17(2), 402 - 7
Activation of botulinum toxins in the absence of nicking; Ohishi I et al.; The derivative toxins purified from cultures of proteolytic strains of Clostridium botulinum types A and F were found to have been only partially nicked but were fully activated . Trypsinization of C . botulinum type B derivative toxin at pH 6.0 resulted in simultaneous activation and nicking, whereas at pH 4.5, activation preceded nicking . The toxin was split by trypsin at pH 6.0 into two fragments with molecular weights of 112, ooo and 57,000 . The toxin contained at least three trypsin-sensitive peptide bonds, one of which was more sensitive than the others at pH 6.0 . These results indicate that activation of botulinum toxins by trypsin or endogenous protease (s) is not a direct result of nicking.

Biochim Biophys Acta, 1977 Jul 22, 493(1), 122 - 31
The low temperature magnetic circular dichroism spectra of iron-sulphur proteins . I . Oxidised rubredoxin; Rivoal JC et al.; Variable temperature magnetic circular dichroism spectra have been measured on oxidised Clostridium pasteurianum rubredoxin . Evidence has been obtained for the presence of two one-electron charge-transfer transitions, sulphur to ferric ion, in the region 15 000 to 28 000 cm-1 . The first moment of the lower energy band is consistent with it being the orbital transition t1 non-bonding sulphur orbital, to the 2 e ferric d-orbital . The magnitude of the spin-orbit coupling constant in the lower excited state has been determined and shown to be small compared with the axial distortion . The splitting of the low energy band observed in the absorption spectrum can therefore be equated directly with the axial distortion of the lowest excited charge-transfer state . Finally, the potential utility of making saturation experiments at very low temperatures has been examined.

Lancet, 1977 Jul 16, 2(8029), 129 - 30
An unusual outbreak of food-poisoning associated with meals-on-wheels; Jephcott AE et al.; An outbreak of food-poisoning after a meals-on-wheels lunch affected 49 persons, 1 of whom died . Bacillus cereus serotype 10 and Bacillus licheniformis were isolated in large numbers from many of the patients including the deceased and from the remains of the meal . Clostridium perfringens (C . welchii) serotype 68, which was isolated from many of the patients and the deceased but not from the food, may also have been responsible . Food kept warm during distribution can produce an ideal environment for bacterial multiplication and similar outbreaks may occur again.

Mikrobiologiia, 1977 Jul-Aug, 46(4), 761 - 6
{Microbiological characteristics of Lake Turali}; Aliverdieva-Gamidova LA; The Turali Lake is a salt-water reservoir . The high salt content of water and ooze determine the microflora of the lake . The total bacterial number is lower in the summer than in the winter when the salt concentration in the lake decreases . The highest number of bacteria and sapropnytes is found in the spring . As a result of the high salt concentration in the water and ooze of the Turali Lake, the following physiological bacterial groups are partly or entirely absent: Azotobacter, Clostridium pasteurianum, hydrocarbon-oxidizing, methanol-producing and sulphate-reducing bacteria.

Mikrobiologiia, 1977 Jul-Aug, 46(4), 689 - 94
{Morphogenesis and function of gas caps on spores of anaerobic bacteria of the genus Clostridium}; Duda VI et al.; Contrary to other bacteria, those belonging to the genus Clostridium have their formation of vacuoles being strictly confined to the stage of spore production . In Clostridium, gas vacuoles are formed in the intraexosporial space, i . e . in the cytoplasm between the exosporial membrane and spore coats . Gas caps, being conical accumulations of gas vacuoles, impart increased floatation to spores and favour therefore their distribution in nature . Consequently, Clostridium species have ecological advantage if they can form gas vacuoles . The function of the exosporium and specific rod-shaped appendices of spore coats is discussed.

J Bacteriol, 1977 Jul, 131(1), 366 - 8
Selenium requirement for the growth of Clostridium sporogenes with glycine as the oxidant in stickland reaction systems; Costilow RN; Clostridium sporogenes was found to have an absolute requirement for selenium to utilize glycine but not proline as oxidant in Stickland-type fermentations . No glycine reductase activity was detectable in cells from media without added selenium . The data indicate that this organism could be used for microbiological assays for very low levels of selenium in certain forms.

Radiology . 1977 Jul;124(1):26.
Clostridium-produced gas gangrene of the colon; Rudikoff JC; A case of radiologically demonstrable gas gangrene of the colon was proved pathologically to be caused by C . perfringens . The case was radiologically indistinguishable from that of bowel infarction . When intramural gas is seen in patients with the symptoms of toxemia but without the bloody diarrhea associated with bowel infarction, infectious gas gangrene should be considered.

Appl Environ Microbiol, 1977 Jul, 34(1), 99 - 101
Proteolytic mutants obtained from Clostridium botulinum type E; Nakane A et al.; Proteolytic mutants were isolated from toxigenic strains of Clostridium botulinum type E after several transfers . When these cultures were plated on blood agar, almost all of the colonies obtained were proteolytic, and there were fewer toxigenic colonies than nontoxigenic colonies . The proteolytic mutants and nonproteolytic original strains were different in their biological properties.

Appl Environ Microbiol, 1977 Jul, 34(1), 23 - 9
Thermal destruction of Clostridium botulinum spores suspended in tomato juice in aluminum thermal death time tubes; Odlaug TE et al.; The heat destruction characteristics of Clostridium botulinum spores suspended in tomato juice and phosphate buffer were determined by the survivor curve method with aluminum thermal death time tubes . Two type A strains of C . botulinum and a type B strain were evaluated . Strains A16037 and B15580 were implicated in outbreaks of botulism involving home-canned tomato products . Strain A16037 had a higher heat resistance than either 62A or B15580 . The mean thermal resistance (D-values) for A16037 in tomato juice (pH 4.2) were: 115.6 degrees C, 0.4 min; 110.0 degrees C, 1.6 min; and 104.4 degrees C, 6.0 min . The mean D-values for A16037 in Sorensen 0.067 M phosphate buffer (pH 7) were: 115.6 degrees C, 1.3 min; 110.0 degrees C, 4.4 min; and 104.4 degrees C, 17.6 min . At each test temperature, the D-values were approximately three times higher in buffer than in tomato juice . The z-value for C . botulinum A16037 spores in tomato juice was 9.4 degrees C, and in buffer the z-value was 9.9 degrees C . The use of aluminum thermal death time tubes in a miniature retort system makes it possible to determine survivor curves for C . botulinum spores at 121.1 degrees C . This is possible because the lag correction factor for the aluminum tubes is only about 0.2 min, making possible heating times as short as 0.5 min.

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