Vibriosis

Zh Mikrobiol Epidemiol Immunobiol, 1977 May, (5), 128 - 9
{Disinfecting effect of fixatives in histoenzymatic analysis of Vibrio cholerae}; Shchedrin VI et al.; Data are presented on the disinfecting action of fixative fluids on the cholera causative agents in the freshly frozen sections from the tissues and organs of animals infected with cholera . The results obtained offered a possibility of drawing a conclusion on the harmlessness of the histoenzymatic studies on the material infected with cholera vibrio.

Zentralbl Bakteriol {Orig A}, 1977 May, 238(1), 66 - 79
Development and use of single "polytropic" diagnostic tubes for the approximate taxonomic grouping of bacteria isolated from foods, water and medicinal preparations; Mossel DA et al.; Single tubes, containing in all instances a bottom layer of a solid medium for detecting mode of attack on glucose, lactose, mannitol or starch and top layers of solid, semi-solid or liquid media allowing assessment of motility, formation of catalase, oxidase, coagulase, indole, hydrogen sulphide, acetyl methyl carbinol or pigments, as required ("polytropic" diagnostic tubes) were developed for the approximate taxonomic grouping of bacteria commonly encountered in foods, water and medicinal preparations . They are designated as: Gram negative diagnostic tubes (GNT), tubes allowing identification of E . coli, following the principle set out by MCKENZIE, TAYLOR and GILBERT (MTGT), diagnostic tubes for Vibrio parahaemolyticus (VPT), open tubes to assess oxidative attack on glucose (OGT), Gram positive diagnostic tubes (GPT), and mannitol plasma tubes (MPT) for the identification of Staph . aureus . In addition a CP tube for the identification of Cl . perfringens is described, the basis of which is the production of H2S from sulphite in the presence of cycloserine, absence of mitality and failure to produce indole at 46 degrees C . All tubes contain intermediate layers to avoid interactions between changes occurring in top and bottom layers . Upon examination of about 600 pure cultures from collections or freshly isolated from foods etc . such tubes have given results which were in agreement with those of classical testing; in a few instances (testing for indole formation) even better results were obtained . The use of such polytropic tubes in identification routes saving much time and effort is outlines.

J Infect Dis, 1977 May, 135(5), 720 - 8
Characterization of the Chinese hamster ovary cell assay for the enterotoxins of Vibrio cholerae and Escherichia coli and for specific antisera, and toxoid; Guerrant RL et al.; The morphologic change in Chinese hamster ovary (CHO) cells that is associated with increased intracellular concentrations of adenosine 3':5'-cyclic phosphate (cyclic AMP) provided a highly sensitive assay for the heat-liable enterotoxins of Vibrio cholerae and Escherichia coli . The similarity between the responses of CHO cells and those of intestinal mucosa to enterotoxins was demonstrated by the inhibition of these effects by the ganglioside galactosyl-N-acetylgalactosaminyl {sialosyl} lactosyl ceramide (GM1), as well as by specific antisera . The antisera against either enterotoxin were more effective in inhibiting the effects of homologous toxin than those of heterologous toxin, but cross-neutralization also occurred in both cases . When choleragenoid, the natural toxoid of cholera toxin, was incubated with the CHO cells, the subsequent effects of the toxin on the morphology of the cells were prevented as were the effects of the toxin on intestinal secretion . However, choleragenoid did not prevent the effects of E . coli toxin . The similarity between the action of enterotoxins on CHO cell morphology and on secretion by the small intestine suggests that there are similar binding sites in these tissues . The potential usefulness of the CHO cell assay in quantitative measurement of antitoxic immunity is thus demonstrated.

Can J Microbiol, 1977 May, 23(5), 630 - 3
Suitability of some enrichment broths and diluents for enumerating cold- and heat-stressed Vibrio parahaemolyticus; Beuchat LR; Vibrio parahaemolyticus cells were injured by chilling and heating, and their recovery was tested in glucose-salt-Teepol broth (GSTB), tryptic soy broth containing 7% NaCl (TSBS), Horie - arabinose - ethyl violet broth (HAEB), and water blue - alizarin yellow broth (WBAY) . Exponential phase cells were more sensitive to cold shock than were stationary phase cells . Exposure of chill-injured V . parahaemolyticus to GSTB and TSBS resulted in 70 to 80% death; about 70% lethality was noted for heat-injured cells inoculated into TSBS . Neither HAEB nor WBAY enrichment media were lethal to stressed cells, although rates of growth were retarded . The 3% NaCl in 0.1 M potassium phosphate (pH 7.0) diluent proved to be most suitable for protecting against inactivation of cold- and heat-injured cells.

Schweiz Med Wochenschr, 1977 Apr 16, 107(15), 545 - 6
{Effect of orally administered pancreatic extract on Vibrio cholerae infection during protein deficiency}; Gyr K et al.; Two groups of 5 and 6 Vervet monkeys respectively were fed a protein-free diet . Both groups of protein depleted Vervets developed a minor decrease of serum and intestinal immunoglobulins, as well as marked impairment of exocrine pancreatic function . Thereafter the groups were challenged with Vibrio cholerae, while one group received purified pancreatic extract by oral administration . The group without pancreatic extract developed severe and significantly longer lasting diarrhea than the group with pancreatic extract . Vibrios were excreted for much longer in the former group . It is concluded that exocrine pancreatic function is involved in the local defence mechanism against cholera during protein deficiency.

Experientia, 1977 Apr 15, 33(4), 518 - 20
Preparation of fixed antigenic, non-oncogenic and protective neoplastic cells; Hakim AA; Transplantable spontaneous mammary adenocarcinoma and sarcoma P 1798 cells were incubated with vibrio cholera neuraminidase (VCN), then fixed cells demonstrated ability to exclude trypan blue dye and to immunized animals and produce cytotoxic sera of high titers . The fixed neuramindase-treated tumor cells became non-oncogenic and protected the host against high doses of fresh untreated homologous tumors.

Dev Biol Stand, 1977 Apr 13-15, 38, 413 - 9
Toxicological studies with therapeutically-applicable Vibrio cholerae neuraminidase; Ronneberger H; Neuraminidase was submitted to a preclinical toxicological investigation in short--and long--term experiments . The acute toxicity in mice, rats, and guinea pigs is low (LD 50 i.v.: mouse 30,000 units/kg, rat 44,000 units/kg, guinea pig 7,700 units/kg; s.c.: mouse, rat, guinea pig more than 30,000 u/kg) . After i.v . injection into dogs, respiration is not at all and blood pressure only slightly altered . Neuraminidase is pyrogenfree in the rabbit test . Assays of antigenicity in guinea pigs and tests on local i.v., i.m., s.c . toxicity in rabbits and guinea pigs showed only moderate toxic signs . Chronicity studies were performed in dogs and rats over 90 days with repeated administration of various amounts of Neuraminidase (dogs 50-2,000 u/kg, rats 500-2,000 u/kg i.v., 26 or 65 injections) . Only in very high dose levels (more than 500 u/kg) Neuraminidase was toxic in dogs (damage of liver, kidney and erythrocytes) . In lower doses in dogs and in rats the clinical, biochemical, and hematological data as well as urinalysis and anatomopathological results give no evidence of systemic or important alterations which would preclude the clinical investigation of the drug.

J Biol Chem, 1977 Apr 10, 252(7), 2412 - 8
Action of intrinsic sialidase of rat brain synaptic membranes on membrane sialolipid and sialoprotein components in situ; Yohe HC et al.; The sialo compounds in the synaptosomal membranes of young rat brain were specifically labeled in vivo by the intracranial injection of radioactive N-acetylmannosamine . More than 95% of the incorporated label was found in glycosidically bound sialic acid . Specific activities of sialic acid in the synaptic membrane gangliosides G71 (monosialo), GD1a (disialo), and GT1 (trisialo) were similar; labeling in GD1b (disialo) was consistently somewhat higher . The highest specific activity of rat brain sialidase was evenly distributed between "small myelin fragment" and synaptosomal membrane fractions, and ouabain-sensitive (Na+, K+)-ATPase also was concentrated in the latter fraction . The greatest amount of bound sialic acid was found in these subcellular fractions having the highest sialidase activity . A microsomal fraction was discovered to contain a small amount of bound sialic acid with a very high degree of radioactive labeling, but no sialidase . Release of sialic acid from the relatively intact membrane preparations by intrinsic membrane-bound sialidase occurred in two recognizable stages . There was a rapid initial release, complete within 30 min, of approximately equal amounts of lipid- and protein-bound sialic acid, corresponding to roughly half of the enzymatically releasable protein-boudn, and somewhat less than one-third of the lipid-bound, sialic acid . The remainder of the membrane sialidase-susceptible sialic acid was released in a second, slower stage . The intrinsic sialidase released 16 +/- 1% of the total sialoprotein and 31 +/- 1% of the total sialolipid sialic acid . Approximately the same amount of sialic acid is releasable from membrane sialolipid by the action of exogenous Vibrio sialidase; almost twice as much is releasable from sialoglycoprotein by this enzyme as compared with the intrinsic membrane sialidase . Each of the various membrane gangliosides appeared to be equally available to the action of the membrane sialidase . The results of this study indicate that both glycolipid- and glycoprotein-bound sialic acid in the synaptic membrane are releasable in situ by the action of the intrinsic synaptic membrane sialidase, and they suggest that this enzyme may act to modulate the physical properties of the membrane . In addition to influencing the rate of hydrolysis of endogenous membrane sialo compounds by intrinsic sialidase, pH had an effect on availability of protein-bound sialic acid . At acid pH, lipid- and protein-bound sialic acid were similarly available, but near neutral pH, gangliosides appeared to be attacked preferentially.

Zh Mikrobiol Epidemiol Immunobiol, 1977 Apr, (4), 59 - 63
{Cholerogenicity of L-forms of cholera vibrios}; Lomov IuM et al.; Dutta and Habbu's method was applied to comparative study of cholerogenic properties of the L-strains of cholera vibrios and their initial bacterial variants . In difference from the initial strains, L-forms of cholera vibrios possessed no cholerogenic properties . A possibility of reversion of the stable L-form of cholera vibrios into bacterial form in vivo was revealed . This reversion was accompanied by the restoration of the cholerogenic properties and the change of the biotype of the cholera vibrio.

J Clin Microbiol, 1977 Apr, 5(4), 444 - 7
Anaerobic vibrio-like organisms cultured from blood: Desulfovibrio desulfuricans and Succinivibrio species; Porschen RK et al.; Two unusual anaerobic vibrio-like organisms were recovered from blood cultures of two patients . One isolate was identified as Desulfovibrio desulfuricans . It appeared to be the cause of a 24-h episode of fever, chills, and profuse perspiration . This is apparently the first documented report of human infection due to this organism . The second isolate was a Succinivibrio species . It has rarely been described as a cause of bacteremia . The clinical significance of the organism remains unclear.

J Infect Dis, 1977 Apr, 135(4), 654 - 8
Sucrose teepol tellurite agar: a new selective indicator medium for isolation of Vibrio species; Chatterjee BD et al.; Sucrose teepol tellurite (STT) agar is a more successful medium in positive-recognition palting procedures than thiosulfate-citrate-bile salts-sucrose (TCBS) agar because STT agar yields higher isolation of typical colonies of Vibrio cholerae, which directly agglutinate in antiserum to V . cholerae, from patients and contacts infected with this organism . STT agar has a simple composition and, like TCBS agar, needs no sterilization . STT agar is highly selective for V . cholerae, nonagglutinating vibrios, and Vibrio parahaemolyticus . Growth of these bacteria from diarrhetic stool, from rectal swabs, and from stock cultures was compared on different media.

J Bacteriol, 1977 Apr, 130(1), 529 - 31
New R plasmid-mediated restriction-modification system of deoxyribonucleic acid conferred by group E R plasmids; Arai T et al.; A new R plasmid-mediated restriction-modification system of deoxyribonucleic acid was identified . This system is specific for group E plasmids which have been detected in unidentified marine Vibrio fish pathogens.

Am J Epidemiol, 1977 Apr, 105(4), 349 - 61
Cholera on Guam, 1974: epidemiologic findings and isolation of non-toxinogenic strains; Merson MH et al.; In August 1974, six cases of cholera occurred on Guam . The index case had severe diarrhea and metabolic acidosis and died from pneumonia on the ninth day of illness; the other five cases had only mild to moderate diarrhea . Fish caught in Agana Bay and home-preserved was found to be the vehicle most likely responsible for the cases . Vibrio cholerae, El Tor Ogawa, was isolated from two patients, the Guam sewerage system, and a river emptying into Agana Bay . V . cholerae, El Tor Inaba, was isolated from the sewerage system, three storm drains imptying into Agana Bay, and Agana Bay . The Ogawa and Inaba isolates differed in their sucrose fermentation and hemolysis reactions, phage type and ability to produce toxin . Although this was the first reported cholera outbreak on Guam, the isolation of differentV . cholerae strains suggested that multiple introductions of V . cholerae had occurred on the island.

Am J Epidemiol, 1977 Apr, 105(4), 344 - 8
Cholera in Portugal, 1974 . II . Transmission by bottled mineral water; Blake PA et al.; During a cholera epidemic, Vibrio cholerae was isolated from two springs which supplied mineral water to a spa and to a commercial water bottling plant . Epidemiologic investigation found that cholera attack rates were 10-fold greater among visitors to the spa than among non-visitors . A subsequent matched-pair case-control study which excluded persons who had visted the spa showed that a history of consumption of the bottled non-carbonated water was significantly more common among bacteriologically confirmed cholera cases than among paired controls.

Am J Epidemiol, 1977 Apr, 105(4), 337 - 43
Cholera in Portugal, 1974.I . Modes of transmission; Blake PA et al.; In April-November 1974, Portugal had a cholera epidemic caused by Vibrio cholerae El Tor Inaba with 2467 bacteriologically confirmed hospitalized cases and 48 deaths . Most of the country was affected, with 17 of the 18 districts reporting cases . V . cholerae was isolated from 42 per cent of shellfish tested during the epidemic, and an epidemiologic study found that a history of consumption of raw or poorly cooked cockles was significantly more common among cholera patients than among paired controls . Water from a spring and a brand of commercially bottled water were also found to be vehicles of transmission of cholera . Although night soil was sometimes used on gardens, consumption of raw fruits and vegetables was not associated with illness.

J Gen Microbiol, 1977 Apr, 99(2), 325 - 31
Transfer and expression of pseudomonas plasmid RP1 in Caulobacter; Alexander JL et al.; This study demonstrates that the host range of Pseudomonas plasmid RP1 includes the genus Caulobacter . Caulobacter was shown to acquire three antibiotic resistance markers located in RP1 . A fourth plasmid marker, susceptibility to an RNA bacteriophage, was not expressed, but could be transferred from Caulobacter to Escherichia coli . The lack of phenotypic expression of the phage marker was manifested by the inability of the phage to adsorb or to produce plaques on Caulobacter transcipients . Matings of Pseudomonas aeruginosa and Caulobacter vibrioides CV6 were carried out in the presence of bacteriophage phi6, a DNA phage that infects and kills only swarmer cells of Caulobacter . No decrease in plasmid transfer in the presence of phage phi6 was detected, suggesting that stalked cells, and not swarmer cells, serve as recipients . Our evidence suggests that transfer of chromosomal segments from Caulobacter may be mediated by plasmid RP1; such segments are not stably maintained.

Infect Immun, 1977 Apr, 16(1), 374 - 81
Reduction of reactivity of Escherichia coli enterotoxins by intestinal mucosal components; Cole HD et al.; Incubation studies involving rabbit and piglet small intestinal mucosal components and Escherichia coli and Vibrio cholerae enterotoxins were conducted at 37 and 4 degrees C . Mucosal homogenate cytosol from rabbits did not significantly alter the reactivities of either cholera enterotoxin (CT) or E . coli labile enterotoxin (LT) . However, mucosal homogenate cytosol from piglets was capable of neutralizing LT, though it had no effect on E . coli stable enterotoxin . LT became bound to piglet and rabbit microvillous membranes at 4 degrees C in the presence of a protective protein . In rabbits, the binding of LT was not dependent upon an intact glycocalyx or free unbound CT-receptors, although some binding was apparently associated with the glycocalyx and CT-receptors . These results indicated the presence of two different LT-receptors in microvillous membranes one being associated with the membrane proper and the other with the glycocalyx . Stable enterotoxin did not bind to in vitro preparations of piglet mucosal components, which is evidence for a different mechanism for inducing intestinal secretion.

Infect Immun, 1977 Apr, 16(1), 328 - 34
Isolation of cryptic plasmid deoxyribonucleic acid from Kanagawa-positive strains of Vibrio parahaemolyticus; Guerry P et al.; Twelve strains of Vibrio parahaemolyticus was examined for plasmid deoxyribonucleic acid (DNA) by dye-buoyant gradient centrifugation . Four Kanagawa-positive strains, all isolated from the same outbreak of gastroenteritis, contained multiple plasmid species of cryptic function . However, three Kanagawa-negative strains and five Kanagawa-positive strains were not found to contain demonstrable plasmid DNA . R-plasmids were successfully transferred from Escherichia coli to V . parahaemolyticus.

Nord Vet Med, 1977 Apr-May, 29(4-5), 199 - 211
An Aeromonas species implicated in ulcer-disease of the cod (Gadus morhua); Larsen JL et al.; Vibrio anguillarum is the most frequently isolated bacterial species involved in ulcer disease in salt water fish . However, an Aeromonas species was very often incriminated in ulcers and septicaemia in cods (Gadus morhua) in Danish coastal areas . These Aeromonas strains were very uniform in their biochemical activities only differing in growth on Simmons citrate agar, and the mean value for the guanine + cytosine content of DNA was 59 per cent with a standard deviation of 0.4 . Primary serological investigations demonstrated a possible identity to O and K antigens . The most characteristic biochemical features of these strains were a gas production from glycerol but not from glucose . They were positive in lysine decarboxylase, did not produce indole, and had a typical fermentation pattern of the glycosides . Negative results were found in H2S production, phosphatase and in the utilization of NH4+ and glucose as the only source of N and C . The evident differences from the species described and subspecies of Aeromonas elucidate the weakness of the existing systems of biotyping . A broader conception of the biochemical spectrum for the individual species of Aeromonas combined with serotyping would seem to be a better system for identification, and also for an epidemiological purpose.

Rev Ig Bacteriol Virusol Parazitol Epidemiol Pneumoftiziol Bacteriol Virusol Parazitol Epidemiol, 1977 Apr-Jun, 22(2), 109 - 13
{Antigenic relations between Vibrio cholerae and Aeromonas hydrophila}; Nacescu N et al.; The antigenic relationships between Vibrio cholerae and the oxidase-positive intestinal bacteria, agglutinated on slides by cholera antiserum, were investigated by the agglutination reaction . Of the 478 oxidase-positive strains studied only strain 209 A, belonging to species Aeromonas hydrophila was selected . Evidence was found of common agglutinogenic fractions between Vibrio cholerae and Aeromonas hydrophila 209 A . Attention is drawn to possible confusions in the preliminary stages of a laboratory diagnosis of cholera.

Med J Aust, 1977 Mar 19, 1(12), 405 - 6
A case of non-agglutinable vibrio gastroenteritis with anuria; Harris AR; A case of non-agglutinable vibrio (Heiberg Group II) gastroenteritis is presented . The patient, an alcoholic, acquired this infection in Australia and recovered fully . The need to use selective vibrio media when diarrhoeal stools are cultured is emphasized.

Science, 1977 Mar 4, 195(4281), 897 - 8
Surface molecules of hematopoietic stem cells: requirement for sialic acid in spleen colony formation; Tonelli Q et al.; Enzymatic treatment was used to test the function of some surface peptides and carbohydrates in hematopoietic spleen colony formation . Proteases and most glycosidases had no effect on spleen colony formation, whereas treatment with Vibrio cholerae neuraminidase reduced colonies by one-half . Intact sialic acid (residues appear to play an important role in colony formation.

Southeast Asian J Trop Med Public Health, 1977 Mar, 8(1), 13 - 8
Study of intestinal immunity against V . cholerae: role of antibody to V . cholerae haemagglutinin in intestinal immunity; Chaicumpa W et al.; Cell-bound haemagglutinin as an adhesive factor of Vibrio cholerae has been partially purified from E1 Tor vibrios using 0.05 M cyclohexylaminopropane sulfonic acid buffer pH 9.5 and gel filtration column chromatography . Rabbits were immunized with the precipitin complexes of the haemagglutinin and its antibody . The antiserum is tested for the protective ability against the oral challenge with Vibrio cholerae strain of which the haemagglutinin has been prepared . The results indicate definite protection of the haemagglutinin antiserum.

Infect Immun, 1977 Mar, 15(3), 704 - 12
Intestinal fluid accumulation induced by oral challenge with Vibrio cholerae or cholera toxin in infant mice; Baselski V et al.; The diarrheal response of orally inoculated infant mice to viable Vibrio cholerae and purified cholera toxin was quantitated by means of a fluid accumulation (FA) ratio . The FA ratio is defined as the gut weight/remaining body weight . FA ratios were determined in relation to time of exposure and dose . Onset of fluid accumulation with viable cells of strains CA401 and 569B occurred 8 h postinoculation and reached a near maximum of 16 h . A dose of 4 x 10(6) colony-forming units of strain CA401 was required for a positive response 16 to 18 h postinoculation . Several other classical cholera strains demonstrated a similar dose-related response . Strain 569B, however, required a 100-fold higher dose to give a positive response . Several mutant cholera strains were decreased virulence in other model systems elicited FA ratios decreased from wild-type values . Onset of fluid accumulation which cholera toxin occurred 6 to 8 h postinoculation and reached a maximum by 10 h . A dose of 0.5 microng was required for a positive response 10 to 12 h postinoculation . The positive response to toxin could be inhibited by preincubation with specific antitoxin.

Zh Mikrobiol Epidemiol Immunobiol, 1977 Mar, (3), 88 - 91
{Immunoglobulins and their specific activity in cholera}; Nechaeva IP et al.; The authors studied the quantitative content of nonspecific immunoglobulins and antibodies of the IgG-, IgA-, and IgM-classes to the O-antigen of the cholera vibrio . There proved to be no marked dynamics of the quantitative immunoglobulins indices of various classes established at periods from the 3rd and the 18th day from the beginning of the disease in persons who sustained cholera . In persons who sustained cholera in the nonendemic focus of the disease serum antibodies were represented chiefly by the IgM-antibodies . The formation of IgA-antibodies in the persons examined differed from the regularities detected for the IgG- and IgM-classes and were characterized by rapidity and short duration . No correlation was found between the immunoglobulin levels of various classes and the serum antibody levels in case of cholera affection.

Zh Mikrobiol Epidemiol Immunobiol, 1977 Mar, (3), 67 - 71
{Cholera vibrio fractions . I . Isolation of alkaline extracts of cholera vibrios and a study of their biological properties}; Rubtsov IV et al.; The authors suggest a well-reproducible method of preparation of a biologically-active fraction of an alkaline extract of cholera vibrio from the strain 569B of Inaba serological type consisting in a preliminary triple removal of endotoxin (O-antigen) of the causative agent from the microbial cell suspension with trichloracetic acid with the subsequent extraction of the microbial mass and neutralization of the extract obtained capable of effective detection of the state of specific sensitization of the organism to cholera vibrio . Preparations of the alkaline extracts (cholera allergen) obtained were standard by chemical composition, they were characterized by a high protein content (up to 77%) and were practically nontoxic . These preparations can be used as a bacterial allergen for the experimental investigations.

Hoppe Seylers Z Physiol Chem, 1977 Mar, 358(3), 391 - 6
Inhibition studies on Vibrio cholerae neuraminidase; Brossmer R et al.; A series of viral neuraminidase inhibitors showing no structural analogy to neuraminic acids have been tested to find whether they are effective inhibitors of V . cholerae neuraminidase, too . Here we report the results obtained with the N-phenyloxamic acid derivatives 2 to 6 (R-NH-CO-COOR'; R = -C6H5NO2, -C6H5OH, -C6H5NH2; R' = -H, -C2H5; see Table 1) and with simple aromatic compounds structurally related to R, i.e . 4-nitroaniline (7), N-acetyl-4-nitroaniline (8), 4-nitrophenol (9), 2,4-dinitrophenol (10), and 4-aminophenol (11) (see Table 2) . The inhibitory effects of 2 to 11 were studied according to the method of Dixon{19} in 0.1m sodium acetate buffer, pH 5.5, 2mM CaCl2, at 37 degrees C using the benzyl-alpha-ketoside of N-acetyl-D-neuraminic acid (1) as a substrate . The compounds 2 to 11 are shown to be competitive inhibitors of the enzymatic hydrolysis of the alpha-ketoside 1 . The competitive inhibition kinetics are supported by the method of Lineweaver and Burk{20} . The inhibition constants (Ki) are found to be in the range of 0.03 to 5.7 mM . The simple aromatic compounds 7 to 11 show higher inhibitory activities than the phenyloxamic acid derivatives 2 to 6 . In addition, significant differences in the Ki values were observed within the two series of inhibitors, whereby those containing a nitro group were most effective.

Hoppe Seylers Z Physiol Chem, 1977 Mar, 358(3), 397 - 400
Selective isolation of Vibrio cholerae neuraminidase using an immobilized 4-(nitrophenyl)oxamic acid; Brossmer R et al.; N-(4-Nitrophenyl)oxamic acid{1} (1) was coupled with Sepharose 4B containing 1,6-diaminohexane as spacer group . This material was used as a specific adsorbent in the purification of Vibrio cholerae neuraminidase . The enzyme was completely retarded and separated from the bulk of the protein when washed with 50mM sodium acetate buffer, pH 5.0 . A stepwise increase of sodium chloride concentration from 1.0 to 2.0M was found to be necessary for a sharp elution of neuraminidase activity . The purification was tenfold, and a recovery of more than 90% was obtained . Neuraminidase is only weakly retarded on a column of 1,6-diaminohexane coupled with Sepharose 4B and is not adsorbed by Sepharose 4B.

Antibiotiki, 1977 Mar, 22(3), 242 - 3
{Effect of a polysaccharide-contaning preparation from El Tor vibrions on the phagocytic activity of leukocytes and on the lysozyme titer}; Sobolev VR et al.; The dynamics of changes in some indices of the natural immunity of mice treated with EL-Tor vibrio preparations in doses of 50 and 500 gemma/mouse was studied . The blood for analysis was taken from the mice 24 hours, 3, 7 and 10 days after the preparation administration . The results of the study showed that administration of the polysaccharide-containing preparations from El-Tor vibrios in doses of 50 and 500 gemma/mouse was accompanied by some shifts in the regulatory mechanisms: suppression of the serum lysozyme activity on the 1st and 3rd days and its stimulation on the 7th and 10th days . The percentage of phagocytosis and its completeness increased . The indices of phagocytosis and its completeness by the respective periods decreased.

J Bacteriol, 1977 Mar, 129(3), 1266 - 71
Formation and function of Vibrio parahaemolyticus lateral flagella; Shinoda S et al.; Formation of the lateral flagella (L-flagella) of Vibrio parahaemolyticus was studied immunologically, using specific antiserum against L-flagella . On solid medium, L-flagella were formed at both high (37 degrees C) and low (25 degrees C) temperatures, although at high temperatures they became dissociated from the cells and decomposed in the medium . L-flagella were not formed in liquid or soft-agar medium . Formation of L-flagella was decreased by lowering the pH of the medium and repressed by transferring the cells from solid medium to liquid medium . Mutants possessing L-flagella but not a polar monotrichous flagellum (M-flagellum) swarmed on solid medium, whereas mutants were grown on solid medium and then transfered to liquid medium, the cells oscillated until they lost L-flagella . It is postulated that L-flagella are locomotive organelles on solid medium and in some cases also in liquid medium, whereas M-flagella are locomotive organelles only in liquid medium.

Transplant Proc, 1977 Mar, 9(1), 801 - 6
Differential effects of T-lymphocyte-derived soluble factor on virgin and primed B lymphocytes in vitro; Friedman H; Cell-free supernatants from mixed leukocyte cultures derived from histoincompatible mouse strains markedly enhanced the in vitro immune response to SRBC by splenocytes from allogeneic mice . The supernatant factor or factors from allogeneic spleen cell cultures appeared to preferentially stimulate antigen-sensitized B lymphocytes, especially those involved in secondary IgG antibody formation . Furthermore, as shown in the present study, the enhancing supernatant factor or factors had no effect on the true primary in vitro immune response to Vibrio cholerae antigen . Normal spleen cells cultured in vitro without cholera vaccine failed to develop antibody-forming cells to this bacterial antigen, despite the presence of the enhancing factor . In contrast, the true secondary immune response to vibrios was markedly enhanced when allogeneic culture supernatants were added at the time of secondary immunization of cholera-primed splenocytes in vitro . Enhancement occurred both for 7S IgG and 19S IgM vibriolytic plaque-forming cells . It appears likely that T lymphocytes present among the allogeneic splenocytes interacting in vitro to histoincompatible antigens release a factor or factors that primarily affect antigen-primed B lymphocytes but also may influence other cells such as macrophages that are important in the immune response to particulate antigens.

Aust Vet J, 1977 Feb, 53(2), 78 - 81
Some causes of wastage in beef cattle artificial breeding; Donaldson LE; Causes of reproductive loss in beef cattle AI programs were inadequate oestrus detection, use of vasectomised bulls, reduced inseminator efficiency, inadequate nutrition and infectious reproductive diseases (vibriosis and IPV) . Nine case histories were presented detailing the symptoms and diagnosis of low reproductive performance.

Infect Immun, 1977 Feb, 15(2), 533 - 8
Evaluation of surface components of Vibrio cholerae as protective immunogens; Eubanks ER et al.; Surface components of a motile Inaba strain (CA401) were removed from washed cells by low-speed shearing . Flagella contaminated with a vesicular material (designated as crude flagella {CFA1) were obtained by differential centrifugation of the shear fluid . Vesicles were obtained from a nonflagellated mutant by the same procedure . Homogeneous small vesicles were obtained in diminished yield from CsCl gradients of CF preparations . Treatment of CF with sodium deoxycholate removed the vesicular material and flagellar sheaths and yielded naked flagella (NF) . The ability of these preparations of passively protect infant mice suckled by CFW mothers that had been immunized at the time of mating was compared, on a dry-weight basis, with commercial vaccine (CV) . Eight-day-old mice were challenged orally with more than 1,000 50% lethal doses of either the homologous or a heterologous (Ogawa Ca411) strain . The most effective immunogen was CF, which provided complete protection at 1 microng against both challenges . CF and vesicles provided 50- to 100-fold greater protection than CV against homologous challenge . With heterologous challenge, vesicles were 10-fold more protective than CV, markedly less protective than CF . The NF offered only slightly greater protection than CV against both challenges . Immunoelectrophoresis revealed an antigen in CF distinct from vesicles, cell wall lipopolysaccharide or NF . This antigen is not present in the nonflagellated mutant and is apparently associated with motility,

Infect Immun, 1977 Feb, 15(2), 360 - 9
Immunoglobulin and specific-antibody synthesis in vitro by enteral and nonenteral lymphoid tissues after subcutaneous cholera immunization; Svennerholm AM et al.; An in vitro culture technique with synthesis of 14C-labeled protein has been used to study immunoglobulin and specific-antibody formation by spleen, mesenteric lymph nodes, Peyer's patches, and small intestine of rabbits, which were immunized twice subcutaneously with Vibrio cholerae lipopolysaccharide (LPS) and enterotoxin; saline-injected rabbits served as controls . Newly synthesized immunoglobulin G (IgG), IgA, and IgM were quantitated by liquid scintillation after their isolation by means of affinity chromatography from columns with immunoglobulin class-specific antibodies coupled to Sepharose . Specific antibodies were similarly measured after purification from gels derivatized with LPS or cholera toxin . The isolated antibodies had full biological activity as studied in protection tests . The immunization increased the overall IgM synthesis in the spleen . It also enhanced the production of IgA and IgG in Peyer's patches and of IgA in intestine . Significant synthesis of radiolabeled antibodies against both V . cholerae LPS and enterotoxin was found in spleen as well as in Peyer's patches of immunized animals . Titration with an enzyme-linked immunosorbent assay (ELISA) showed significant levels of IgG as well as IgA antibodies in incubation medium from all the studied tissues, whereas specific IgM was only found for spleen and mesenteric lymph nodes . Simultaneous tissue incubations at 37 degrees C and in an ice bath indicated that the major part of the antibodies registered with the ELISA represented de novo synthesis.

Zentralbl Bakteriol {Orig A}, 1977 Feb, 237(1), 65 - 71
Experimental Toxigenicity of NAG Vibrios; Draskovicova M et al.; Some strains of NAG vibrios isolated from the stool of patients with diarrhoeal disease as well as from surface water caused an accumulation of fluid in the ligated rabbit gut loop . 5-fold concentrated sterile culture filtrates of some strains were found positive in this test as well . The volume of the accumulated fluid in gut loops injected with live cultures as well as with concentrated culture filtrates was apparently smaller than the volume accumulated after injection of non-concentrated V.cholerae culture filtrates . This points to the fact that the NAG vibrio strains belong to weaker producers of enterotoxin than the cholera vibrios . The culture filtrates of all investigated strains contained the skin toxin which was of increased vascular permeability in the skin of rabbits . Besides this, a hemorrhagic effect was found in the filtrates . The skin toxin of NAG vibrios appears to be more heat resistant than the toxin of cholera vibrios . The presence of the skin toxin in culture filtrates, however, does not correlate with the enteropathogenicity of NAG vibrio strains.

J Bacteriol, 1977 Feb, 129(2), 1121 - 8
Purification of flagellar cores of Vibrio cholerae; Yang GC et al.; A procedure is described for the purification of the cores of flagella sheared from Vibrio cholerae . V . cholerae is a monotrichous organism whose flagellar core (FC) is enclosed within a sheath . The purification procedure consists of several cycles of differential centrifugation and cesium chloride density-gradient ultracentrifugation in the presence of a neutral detergent, Triton X-100 . Purity of the FC preparations is assessed by electron microscopy, polyacrylamide gel electrophoresis, and chemical analysis . The purified FC preparations are devoid of flagellar sheaths and free from detectable cell wall and cytoplasmic contamination . Antibody prepared in rabbits against purified FC reacts with the flagellum of intact V . cholerae or purified FC as seen by ferritin-labeled antibody studies . Purified FC is composed of a single protein subunit with an estimated molecular weight of 45,000 g/mol and a density of about 1.3 g/cm3.

J Lab Clin Med, 1977 Feb, 89(2), 367 - 77
Functional and metabolic properties of human asialofibrinogen; Martinez J et al.; The role of sialic acid in the functional and metabolic properties of purified human fibrinogen was investigated . Fibrinogen treated with Vibrio cholerae neuraminidase released 90 percent of its sialic acid without evidence of proteolysis, as indicated by the presence of intace A alpha, B beta, and gamma chains on sodium dodecylsulfate (SDS)-polyacrylamide gels of the reduced asialoprotein . The thrombin and Reptilase clotting times of human asialofibrinogen were shortened compared to those of normal fibrinogen . Fibrinopeptide release was normal in rate and amount, but asialofibrin monomer aggregation was increased at both low and high ionic strength . Similarly, the asialo-derivative of fibrinogen Philadelphia (functionally characterized by impairment of fibrin monomer aggregation) demonstrated shortening of its thrombin and Reptilase times and improvement in its monomer aggregation especially at high ionic strength . Asialofibrin showed a normal capacity to form cross-linked fibrin as demonstrated by normal gamma-chain dimerization and alpha-chain polymerization . Simultaneous metabolic studies of human normal fibrinogen and asialofibrinogen in rabbits revealed only a modest decrease in the half-life of the asialoprotein compared to the intact protein, with no preferential uptake of the asialo-derivative by the liver . Control studies with rabbit normal fibrinogen and asialofibrinogen in rabbits revealed the same modest difference in half-life . Thus, asialofibrinogen clots faster due to enhancement of its monomer aggregation, has a normal capacity to form cross-linked fibrin, and does not differ significantly in its metabolic properties from normal fibrinogen . The possible influence of sialic acid in the functional abnormality of some congenital dysfibrinogenemias is discussed.

Infect Immun, 1977 Feb, 15(2), 539 - 48
Use of fluorescent antibody in studies of immunity to cholera in infant mice; Guentzel MN et al.; Infant mice 8 days of age were infected orally with virulent, motile, classical or El Tor strains of Vibrio cholerae and with nonmotile mutants of low virulence derived from the same strains . At intervals of 8 and 12 h postinfection, frozen thin sections of the ileum were prepared, stained with fluorescein isothiocyanate-labeled rabbit anti-vibrio antibody, and examined with the fluorescence microscope . The motile organisms were present in larger numbers, especially at 12h, and had penetrated the intervillous spaces and crypts of Lieberkuhn more completely than nonmotile vibrios . Dilution counts were made on various regions of the intestines of infant mice challenged orally 12 h previously with either motile or nonmotile strains of V . cholerae . Greater numbers of organisms were found, especially in the upper intestinal regions, when motile organisms were used . Low numbers of vibrios, limited mostly to the lumen, were seen in the ileum of infant mice infected with motile organisms when the infants were the offspring of mothers that had been immunized with crude flagellar vaccine or a vesicular preparation derived from the vibrio cell surface . The distribution of vibrios in this case was similar to that found in infected infants of unvaccinated mothers challenged with nonmotile organisms . Motility appears to enable the bacteria to better populate the upper regions of the intestinal tract and to avoid the washing effects of secretions and peristalsis . Antibacterial immunity may function, at least in part, by making it impossible for motile vibrios to accomplish this widespread distribution within the ileum.

Folia Microbiol (Praha), 1977, 22(5), 402 - 9
Effect of rifampicin on tryptophanase induction in normal and rifampicin-resistant Vibrio el tor; Sinha AM et al.; The induction of tryptophanase was not affected by rifampicin in the rifampicin-resistant mutant of Vibrio el tor while the antibiotic inhibited the induction of tryptophanase in the normal strain at level of ribonucleic acid and protein synthesis.

Microbiol Immunol, 1977, 21(5), 243 - 54
Toxin isolation from a Kanagawa-phenomenon negative strain of Vibrio parahaemolyticus; Sochard MR et al.; Production of a toxin by Vibrio parahaemolyticus Kanagawa-phenomenon negative strains was examined . Ammonium sulfate fractions of broth culture filtrates were dialyzed, concentrated by lyophilization, and tested for toxic effects by mouse intraperitoneal injection . One fraction, which we think is a toxin, was isolated from a broth culture filtrate of V . parahaemolyticus FC 1011 (a Kanagawa-phenomenon negative strain) and consitently produced lethal effects in mice at high concentrations and diarrhea in lower concentrations . The toxin was assayed for mouse LD50 and ability to produce diarrhea via forced feeding in mice . V . parahaemolyticus FC 1011 toxin was found to be protein, to be inactivated by heat or trypsin hydrolysis, and to produce positive skin permeability reactions in rabbits . However, it failed to induce fluid accumulation in ligated ileal loops in rabbits.

Ann Microbiol (Paris), 1977 Jan, 128A(1), 35 - 9
Amino sugar contents and phage inactivating properties of lipopolysaccharide from cholera and El Tor vibrios; Maiti M et al.; Cholera and El Tor lipopolysaccharide (LPS) were identical in respect of chemical composition except that the hexosamine content was significantly lower and galactosamine was absent in El Tor LPS . Mukerjee's group IV cholera phage phi149 was inactivated by cholera LPS but was resistant to El Tor LPS.

Jpn J Physiol, 1977, 27(1), 43 - 56
Effect of hemolysin produced by Vibrio parahaemolyticus on membrane conductance and mechanical tension of rabbit myocardium; Seyama I et al.; The mechanisms of the action of hemolysin extracted from Vibrio parahaemolyticus in the S-A node and right atrium cells of rabbit were studied by means of the single sucrose gap and isometric tension recording methods . Hemolysin caused the membrane to depolarize reversibly without affecting the action potential generating mechanism . Lowering of {Na+}o inhibited membrane depolarization in the presence of hemolysin while the readmission of normal Tyrode solution induced depolarization . Tetrodotoxin (TTX) barely antagonized the depolarizing action of hemolysin but slowed the rate of development of depolarization . Therefore, this depolarization is considered to be primarily due to the increase in conductance to Na which TTX may not block . The dose-response relationship was obtained by measuring a change in membrane resistance . The concentration necessary to yield one-half of the maximum reduction of the membrane was determined to by 7.5 micrograms/ml . Accumulation of Na within the cell may be responsible for an increase of twitch tension observed during the action of a low concentration of hemolysin . On the other hand, a higher concentration of hemolysin seemed to promote exchange of intracellar Na with extracellular Ca, especially when the Na concentration of the perfusing solution was reduced, and led to stronger contracture.

Zh Mikrobiol Epidemiol Immunobiol, 1977, (1), 88 - 90
{Pathogenic and antigenic properties of NAG-vibrios isolated from man and environmental objects}; Andrusenko IT et al.; A study was made of the pathogenic properties of NAG-vibrios on various experimental animals (917 guinea pigs, 609 nursling rabbits, and 203 rabbits aged from 20 to 24 days) . The results obtained were compared with the reference to definite serological groups . Pathogenic NAG vibrios proved to be encountered in all the serological groups studied . The presence of pathogenic NAG-vibrios was mostly noted among the cultures belonging to the 2nd, 5th, 8th and 18th serological groups . The majority of serological groups isolated from man were also revealed among the cultures obtained from water.

Zentralbl Bakteriol {Orig B}, 1977 Jan, 164(1-2), 119 - 20
{Vibrio parahaemolyticus in mussels and in silt in the southern central baltic sea (author's transl)}; Zaleski S et al.; Because the previous studies of 541 fish of the Southern Central Baltic Sea area gave negative results for the presence of Vibrio parahaemolyticus, 150 shellfish and 40 mud samples of the same origin were tested this time . In none of them presence of Vibrio parahaemolyticus nor Vibrio alginolyticus has been stated, though.

Appl Environ Microbiol, 1977 Jan, 33(1), 10 - 4
Enteropathogenicity of Vibrio parahaemolyticus in the ligated rabbit ileum; Brown DF et al.; The enteropathogenicity of Vibrio parahaemolyticus was investigated by contrasting the effects of whole cells, cell fragments, cell-free preparations, and media constituents injected into rabbit ileal loops . Three of 20 cultures utilized were Kanagawa-negative strains from seawater and sea fish . The remaining 17 cultures included both Kanagawa-positive and -negative strains from Japanese victims of gastroenteritis . Broth culture filtrates concentrated 10-fold by dialysis against 30% Carbowax were unreactive, whereas lyophilized filtrates, regardless of Kanagawa type, as well as all sterile broth preparations containing greater than or equal to 5% NaCl gave positive reactions in the rabbit gut . In contrast, crude lysates derived from broth cultures of Kanagawa-positive strains caused loop dilatation; lysate supernatants were unreactive . Lysates of cells washed from brain heart infusion agar were more reactive than lysates from Trypticase soy agar-grown cells . When agar-grown cell lysates prepared by disruption in saline were dialyzed against distilled water, they were devoid of gut reactivity . Reactivity was restored in dialysands resuspended in saline and in dialysates concentrated 10-fold . The agar-grown cell lysates exhibited Kanagawa-type hemolysis . Our data support the conclusion that the rabbit loop reactivity observed with lyophilized, cell-free culture filtrates may result from excessively elevated NaCl concentrations, and that a toxic factor associated with large-cell particles may be dialyzable, depends on saline for expression, and resembles the Kanagawa hemolysin.

J Gen Microbiol, 1977 Jan, 98(1), 77 - 86
The effect of anaerobiosis and bile salts on the growth and toxin production by Vibrio cholerae; Fernandes PB et al.; Environemntal conditions which might be present in the human intestinal lumen, such as anaerobiosis, a temperature of 37 degrees C and the presence of bile salts, were examined for their effects on the growth and toxin production by Vibrio cholerae strains 569B and B1307 in Syncase and in peptone water media . Using aerobic conditions at 30 degrees C, which are commonly used for enterotoxin production, toxin (5 mug ml(-1)) and pleomorphic cells were detected during the exponential phase of the growth cycle . When the incubation temperature was raised to 37 degrees C, no toxin ( less than 0-1 mug ml(-1)) and no pleomorphic forms were found . In cultures incubated anaerobically at 30 or 37 degrees C, the organisms grew poorly, forming pleomorphic cells which lysed after the cultures reached a maximum turbidity at 640 nm of 1-45 at 12 h . Toxin (2-5 mug ml(-1)) was present at 12, 24 and 48 h . When 0-1% sodium deoxycholate was incorporated into the culture medium, growth was inhibited under aerobic conditions at 30 and 37 degrees C . At 30 degrees C under aerobic conditions and at 37 degrees C under anaerobic conditions, the toxin yield was not significantly affected by the presence of sodium deoxycholate; but at 37 degrees C under aerobic conditions, sodium deoxycholate caused an increase in the toxin yield (5 mug ml(-1)) due to the release of cell-bound toxin.

Chemotherapy, 1977, 23(2), 121 - 8
Trimethoprim-sulphamethoxazole in the treatment of cholera . Comparison with tetracycline and chloramphenicol; Pastore G et al.; 67 of the bacteriologically proved adult acute cholera patients have been examined in order to evaluate the efficacy of TM-SMX in comparison with tetracycline and chloramphenicol in the eradication of Vibrio cholerae from stools . Our results demonstrated that all three drugs sterilized the stools of all patients within 3 days with the exception of one case of TM-SMX's group, which had negative culture stools after 4 days . On the basis of our experience it can be emphasized that TM-SMX can support chloramphenicol and tetracycline in the antibacterial treatment of cholera with the advantage that the drug is efficacious with daily administrations.

Cancer Res, 1977 Jan, 37(1), 95 - 101
Effectiveness of neuraminidase in experimental immunotherapy of two murine pulmonary carcinomas; Alley CD et al.; The effects of direct intratumoral inoculation with Vibrio cholerae neuraminidase and inoculation of tumor-bearing mice with tumor cells incubated with neuraminidase in vitro were studied in C57BL/6 X DBA/2 F1 mice bearing s.c.-transplanted, methylcholanthrene-induced pulmonary squamous cell or Lewis lung carcinomas . The growth of the squamous cell tumor was more greatly inhibited by both treatments than was the Lewis lung tumor . In the squamous cell tumor-bearing mice, both modes of neuraminidase treatment depressed tumor growth by approximately 80% . However, 20% of the mice in the group treated with the neuraminidase-incubated squamous cell vaccine and 10% of those treated intratumorally underwent total tumor regression and developed specific immunity to the squamous cell tumor . although the growth rate of the Lewis lung tumor was suppressed by both types of treatment, the direct intratumoral neuraminidase treatment group underwent a greater depression in tumor growth (73 versus 42%) . A possible explanation of the different results of the two treatments in squamous cell and Lewis lung tumor systems may be based on tumor etiology and cellular composition.

Scand J Immunol, 1977, 6(12), 1345 - 9
Boosting of secretory IgA antibody responses in man by parenteral cholera vaccination; Svennerholm AM et al.; The occurrence of specific antibodies to Vibrio cholerae lipopolysaccharide in serum, milk, and saliva of Pakistani women from a very low socioeconomic group was studied before and after a single subcutaneous cholera vaccination . Before immunization all women had low levels of specific antibodies in serum, primarily of IgM class, and in many cases cholera IgA angibodies were found in milk and saliva as well, indicating earlier natural exposure . The vaccination consistently induced a marked rise in serum antibody titer, and notably also produced significant titer increases in 70% of the milk and in 45% of the saliva samples . Whereas the serum antibodies induced were predominantly of the IgG class, secretory IgA was responsible for most of the titer increase in the secretions . The results indicate that parenteral cholera vaccination can boost local secretory IgA antibody responses in intestinally primed individuals.

J Surg Oncol, 1977, 9(6), 527 - 40
The combined effect of radiotherapy and neuraminidase-treated tumor cells on 3-methylcholanthrene-induced fibrosarcoma; So SK et al.; Active immunotherapy with tumor cells treated in vitro with Vibrio cholerae neuraminidase (VCN) plus mitomycin C augments the antitumor effects of local x irradiation in the treatment of firmly established methylcholanthrene-induced fibrosarcoma, MC-43, in syngeneic C3H/HeJ female mice . In most experiments, the inhibition of tumor growth was greater when VCN-treated tumor cells were combined with local irradiation than could be achieved with VCN-treated tumor cells or local irradiation alone . Even in those experiments in which the immunotherapeutic effect of VCN-treated cells was negligible, the combination of radiotherapy and immunotherapy appeared to be greater than irradiation alone . Similarly, total permanent regression of established tumors occurred more frequently after combined therapy than after immunotherapy or radiation therapy alone.

Vet Med Nauki, 1977, 14(3), 54 - 60
{Studies of the etiology of infectious abortion in cows in Ruse District}; Kolev V et al.; A total of 28,159 blood serum samples have been examined for brucellosis, 87--for leptospirosis, 84--for toxoplasmosis, 554--for the presence of rickettsii and neoreckettsii, and 193--for total protein content, albumin, and protein fractions, taken from cows in the course of the years on 40 dairy farms in the district of Rousse . Bacteriologically were examined 349 aborted fetuses, 178 samples of estral secretion, 57 placentae, and 1002 samples of washings and seminal fluid from bulls for vibriosis . Bacteriologic investigations were also carried out on 383 fetuses for salmonellosis, listeriosis, and colibacteriosis, and virologic studies of 398 placentae and parenchymal organs of fetuses . Twenty-eight fetuses were studied parasitologically for toxoplasmosis, and 118 blood smears were examined for hemosporidiosis . It was established that most important in the etiology of the infectious abortions in cows in the district of Rousse were Vibrio organisms . They were found in 36.9 per cent of the studied bovine fetuses and in 24.4 per cent of all materials examined bacteriologically . Only 4 cases were noted of neorickettsii from fetuses and placentae . Serologically, there were 23.6 per cent cases positive for neorickettsiosis, and 12.6 per cent--for Q fever; 56.4 per cent of the investigated sera proved positive for leptospirosis . In 0.8 per cent of the examined fetuses there was Escherichia coli . Abortions in cows were also due to anaplasmosis and francaillelosis; 14.2 per cent of the investigated respective sera were positive for toxoplasmosis . On the days of abortions the amount of total protein was 7.13 mg%, that of albumins--36%, and of globulins--64% . It is considered imperative to elucidate the cause of abortions of infectious nature in cows in connection with the establishing of cattle-breeding complexes.

Arch Invest Med (Mex), 1977, 8(2), 113 - 22
{Immunogenicity of L5178Y cells modified by different reagents}; Gomez-Estrada H et al.; Lymphoma L5178Y cells were treated with neuraminidase of Vibrio cholerae, potassium iodine, dithiotreitol (DTT), mercaptoethanol, glutaraldehyde, iodoacetamide, merthiolate, sodium periodate, urea, papaine, trypsine and EDTA, to increase immunoreaction in tumor cells . Mice were immunized with modified tumor cells every week for one month . Thereafter non modified tumor cells were transplanted to previously immunized mice . Only the immunization with neuraminidase-treated cells rejected the tumor . Although the immunization with cells treated with potassium iodine, DTT and mercaptoethanol did not reject tumor, prolonged significantly span of life . The other reactives had neither effect on tumor rejection nor on span of life.

Acta Microbiol Acad Sci Hung, 1977, 24(1), 33 - 9
Cross-neutralization reactions of Escherichia coli and Vibrio cholerae enterotoxins as studied by rabbit skin inoculation test; Singh A et al.; Neutralization of enterotoxins of Vibrio cholerae 569 B and Escherichia coli 10407 by antitoxins to V . cholerae 569 B, E . coli 334, 408-3 and 10407 was studies by intradermal inoculation test in the rabbit . Neutralization of V . cholerae enterotoxin by homologous as well as heterologous antisera of E . coli was observed, except that there was no neutralization of the enterotoxin by antiserum to E . coli 408-3 enterotoxin . Neutralization of E . coli enterotoxin to a varied extent by homologous as well as all heterologous antisera, including that of V . cholerae 569 B antitoxin, was also observed.

Chemotherapy, 1977, 23(5), 345 - 55
Identification and treatment with a long-acting sulphonamide of 15 carriers of Vibrio cholerae El Tor serotype Ogawa, during the 1973 cholera epidemic in Naples; De Lorenzo F et al.; During the cholera epidemic in 1973, nine contact carriers of Vibrio cholerae El Tor serotype Ogawa were identified in a family from Naples as well as six others coming from different parts of the city or its surroundings . All the subjects were admitted to the quarantine ward of the 'Cotugno Hospital' (Naples) in which they remained continuously for 14 or 16 days . During this time these bacteriologically confirmed carriers were treated orally with a long-acting sulphonamide (sulphamethoxypyrazine, Kelfizine) . Coprocultural examinations carried out on each patient on average 7 times after the treatment over a period of 6 months, proved the absence of V . cholerae . Serological tests also showed high levels of agglutinating and vibriocidal anticholera Ogawa antibodies in two unvaccinated carriers.

Arkh Patol, 1977, 39(2), 10 - 7
{Interaction of cholera vibrios and escherichiae with the intestinal epithelium (histologic and electron microscopic study)}; Polotskii IuE et al.; Histological and electron-microscopic investigations were carried out of ligated small intestinal loops of 147 rabbits within 24 hours after challenge with cholera vibrios El Tor, different enterotoxigenic E . coli, choleragen and E . coli enterotoxins . These ogranisms attached themselves to enterocytes and multiplied on the surface of the unaltered intestinal epithelium . Thereafter, a greatly pronounced secretion of enterocytes, rather than inflammation, developed, like after the exposure to sterile enterotoxins of E . coli and choleragen, the lumen of isolated rabbit gut loops was overfilled with great amount of fluid discharged . At later period dystrophic alterations in enterocytes also appeared . They were partly associated with excessive secretion, but the severest of them (including even necrotic foci) were caused by the compression of the isolated gut loops epithelium with fluid . The first and obligatory stage of pathogenesis of cholera and cholera-like escherichioses was apparently the attachment of the causative agent to enterocytes and its multiplication on the intestinal epithelium surface, followed by hypersecretion induced by enterotoxin effect.

Infect Immun, 1977 Jan, 15(1), 132 - 7
Induction of Escherichia coli and Vibrio cholerae enterotoxins by an inhibitor of protein synthesis; Levner M et al.; Enterotoxigenic Escherichia coli or Vibrio cholerae 569B (Inaba) grown in the presence of the antibiotic lincomycin, an inhibitor of protein synthesis, produced elevated levels of heat-labile enterotoxin or choleragen, respectively, as assayed by both vascular permeability factor and capacity to elicit fluid accumulation in rabbit ileal loops . This induction of enterotoxin did not reflect either a coupling of lincomycin resistance with increased enterotoxigenicity or an effect of lincomycin on cellular release of enterotoxin, since spontaneously isolated lincomycin-resistant mutants of both E . coli and V . cholerae still required lincomycin for induction, and large increases in E . coli permeability factor activity were found intracellularly as well as extracellularly . After the period of exponential growth, E . coli became refractory to induction by lincomycin, although most of the induced enterotoxin activity appeared only after this period . No increase in copy number of the enterotoxin plasmid in E . coli 711 (P307) was found in induced cells by analysis of deoxyribonucleic acid reassociation kinetics . These and other data suggest that synthesis of enterotoxin, or at least its accumulation, is normally limited by cellular factors whose synthesis is preferentially inhibited by lincomycin . A possible connection between this phenomenon and lincomycin-associated diarrhea is considered.

Infection, 1977, 5(4), 211 - 3
The effect of attapulgite and charcoal on enterotoxicity of Vibrio cholerae and Escherichia coli enterotoxins in rabbits; Drucker MM et al.; Vibrio cholerae and certain strains of Escherichia coli produce heat-labile enterotoxins which play a significant role in the pathogenesis of the intestinal disease . Activated attapulgite, a heated magnesium aluminum silicate, was previously shown to prevent the toxic effects of endotoxin . The present study has revealed that this drug inhibits the toxic effects of cholera and E . coli enterotoxins in the intestinal loop of rabbits, when toxin and attapulgite are pre-incubated prior to injection . Up to 50 to 100 minimal effective doses are inhibited . Attapulgite is effective also when injected separately, albeit simultaneously, into the intestinal loops, but not when administered after the toxin . Since supernates of toxinattapulgite mixtures are non-toxic, it is postulated that attapulgite acts by adsorption and that the attached enterotoxin is no longer toxic to the rabbit intestine . The previously reported effect of charcoal on V . cholerae enterotoxin paralleling that of attapulgite, was confirmed . In contrast to the effects of these absorbents on isolated toxin, both failed to prevent enterotoxicity in the rabbit model of an enterotoxin-producing strain of E . coli.

Arch Virol, 1977, 54(4), 299 - 305
Cell receptors for paramyxoviruses; Wassilewa L; Treatment of chick embryo fibroblasts, calf kidney and BHK cells for 30 minutes with the enzyme neuraminidase from Vibrio cholerae causes an enhancement of the per cent of attached NDV virions . This enhancement does not depend on the multiplicity of infection . The quantity of spontaneously eluted and cellbound virus is two times greater than the quantity of the same virus derived from control cells . N-acetyl-neuramin lactose inhibits the effect of Vibrio cholerae neuraminidase . After prolonged action of this enzyme, the quantity of adsorbed NDV diminishes . Treatment of the same cells with neuraminidase from influenza virus decreases the per cent of adsorbed NDV with respect to controls . The other paramyxovirus--bovine parainfluenza 3 virus adsorbs also more intensively on cells treated with Vibrio cholerae neuraminidase . It is suggested that partial hydrolysis of NANA molecules causes a rearrangement of the cell surface charged groups and thus allows a more effective contact between paramyxoviruses and the cell.

Folia Haematol Int Mag Klin Morphol Blutforsch, 1977, 104(4), 538 - 57
{Studies on the morphology of blood of the European eel (Anguilla agnuilla) . III . Studies on monocytic and lymphoid cells}; Kreutzmann HL; Investigations concerning the monocytic and lymphocytic cell series were carried out on the peripheral blood and dabbings of the kidney, the spleen and intestines of healthy eels (anguilla anguilla), and eels taken ill with vibrio anguillarum . By means of histological, cytochemical, phaseoptical, and electronmicroscopical examinations independent development lines of the monocytes and lymphocytes were established . The place of origination for the monocytes was found to be the kidney, for the lymphocytes the spleen and the intestines additionally . The evidence of the cytochemical assay of enzymes for the differentiation of these blood cells and their cellular systems is primarily dependent on the distribution of the activity of the unspecific esterase, the naphthol-AS-D-chloroacetate-esterase, the peroxidase and the alkalic phosphatase . Macrophages were - on the basis of enzymocytochemical results - seen as derivatives of the monocytes . Plasma cells and lymphoid cells are transformation forms of the lymphocytes . Our results emphasize the significance especially of cytochemistry for the hematological diagnostics of fish diseases.

Biull Eksp Biol Med, 1976 Dec, 82(12), 146 - 9
{NAG-infection in grass frogs (Rana temporaria) subjected to hypothermia}; Avtsyn AP et al.; Rana temporaria kept under hypothermic conditions approaching anabiosis were inoculated with NAG-vibrios and examined clinically, bacteriologically, histologically, and electron microscopically . Oral inoculation of hypothermic frogs with NAG-vibrios resulted in 18 to 24 hours in the development of acute NAG-infection resembling the cholera-like syndrome, and characterized by general intoxication and local enteropathogenic effects . NAG-vibrios persisted in the frog gastrointestinal tract for a long time after the cessation of the acute period of the disease.

Biken J, 1976 Dec, 19(4), 139 - 50
Chemical structure of the peptidoglycan of Vibrio parahaemolyticus A55 with special reference to the extent of interpeptide cross-linking; Kato K et al.; The chemical structure of the cell wall peptidoglycan of Vibrio parahaemolyticus A55 was studied . Estimation of cross linkages between peptide subunits in the peptidoglycan by dinitrophenylation showed that about 30% of the total 2,6-diaminopimelic acid (A2pm) residues were involved in cross linkages . The presence of interpeptide bridges was also demonstrated by isolating bisdisaccharide peptide subunit dimers from Chalaropsis muramidase digests of the cell wall peptidoglycan by gel filtration followed by ion-exchange column chromatography, although most of the building blocks obtained were uncross-linked disaccharide peptide monomers . The chain length of a glycan moiety of the peptidoglycan obtained by treatment with the L-11 enzyme and gel filtration of the digest was also studied . The chain length varied from 7 to 44, but 30% of the glycan fragments had muramic acid at the reducing end and a chain length of 28 to 44 . In conformity with the above structural study it was demonstrated that a particulate enzyme fraction obtained by differential centrifugation of a sonicated preparation of V . parahaemolyticus catalyzed a penicillin-sensitive transpeptidation reaction, using UDP-MurNAc-14C-pentapeptide and UDP-GlcNAc as substrates.

Bull Schweiz Akad Med Wiss, 1976 Dec, 32(4-6), 223 - 32
Sequence of events in the activation of adenylate cyclase by cholera toxin; Kohler T et al.; The toxin of Vibrio Cholera causes fluid secretion from the small intestine by stimulation of adenylate cyclase and elevation of intracellular cyclic AMP concentrations . The toxin is a protein composed of subunits responsible for binding to cell membranes and a subunit responsible for the activation of adenylate cyclase . The binding subunits (B) are non-covalently bonded to the active subunit (A) . The latter is composed of two polypeptides A1 and A2 linked by a disulphide bridge . Exposure of the intestine to toxin results in rapid binding to the brush border membrane . Thence follows a gradual increase in adenylate cyclase activity, and stimulation of electrolyte and fluid secretion . Enzyme localization studies show that the brush border does not contain adenylate cyclase . Thus the stimulation of adenylate cyclase by toxin which interacts with the brush border must be indirect . From recent studies it seems that an activator of adenylate cyclase can be found in cytosol from toxin-treated cells . Incubation of toxin with cytosol or dithiothreitol results similarly in the formation of an activator . Preincubation of toxin with cytosol results in more rapid activation of adenylate cyclase in liver cell membranes than direct addition of cytosol and toxin . Preincubation of cholera toxin for activation, by cytosol, is presumed to be due to the splitting of the disulphide band between the A1 and A2 components of the active subunit . The stimulatory ability resides in A1 and both A1 and NAD are required for the activation of adenylate cyclase . The toxin-stimulated adenylate cyclase has similar characteristics to the enzyme stimulated by non-hydrolysable analogs of GTP such as guanylylimidodiphosphate (GppNHp) . Stimulation by either cholera or GppNHp is irreversible, the responses to catecholamines are enhanced and the enzyme can be solubilized in the activated state.

Appl Environ Microbiol, 1976 Dec, 32(6), 792 - 8
Sublethal heat stress of Vibrio parahaemolyticus; Emswiler BS et al.; When Vibrio parahaemolyticsu ATCC 17802 was heated at 41 degrees C for 30 min in 100 mM phosphate-3% NaCl buffer (pH 7.0), the plate counts obtained when using Trypticase soy agar containing 0.25% added NaCl (0.25 TSAS) were nearly 99.9% higher than plate counts using Trypticase soy agar containing 5.5% added NaCl (5.5 TSAS) . A similar result was obtained when cells of V . parahaemolyticus were grown in a glucose salts medium (GSM) and heated at 45 degrees C . The injured cells recovered salt tolerance within 3 h when placed in either 2.5 TSBS or GSM at 30 degrees C . The addition of chloramphenicol, actinomycin D, or nalidixic acid to 2.5 TSBS during recovery of cells grown in 2.5 TSBS indicated that recovery was dependent upon protein, ribonucleic acid (RNA, and deoxyribonucleic acid (DNA) synthesis . Penicillin did not inhibit the recovery process . Heat-injured, GSM-grown cells required RNA synthesis but not DNA synthesis during recovery in GSM . Chemical analyses showed that total cellular RNA decreased and total cellular DNA remained constant during heat injury . The addition of {6-3H}uracil, L-{U-14C}leucine, and {methyl-3H}thymidine to the recovery media confirmed the results of the antibiotic experiments.

Pathol Biol (Paris), 1976 Dec, 24(10), 685 - 9
{Practical interest of a micromethod for neuraminidase treatment of lymphocytes in transplantation immunology (author's transl)}; de Mouzon A et al.; Vibrio cholerae neuraminidase treatment increases the cell sensitivity to complement and antibodies cytotoxic action . This property can be applied to the microlymphocytotoxicity technic for antibodies study in dialysed and kidney transplanted patients and for pretransplantation cross-matches . The enzymatic treatment usually employed needs a great deal of lymphocytes submitted in a second step to antibodies cytotoxic action . But this method appeared difficult to be routinely applied . We developed a simpler method consisting in treating only the lymphocytes needed to perform the test, on the reaction plate itself . This method gives the same results as the classical enzymatic treatment: the intensity of the weakly positive reaction is strongly increased, after pretreatment of cells; in certain cases, it allows one to detect cytotoxic antibodies not revealed without neuraminidase . These antibodies can be related (or not) to HLA system but practically the more important fact is the ability of this method to reveal an eventual incompatibility.

J Immunol, 1976 Dec, 117(6), 2150 - 7
Effect of Vibrio cholerae neuraminidase on the generation of cell-mediated cytotoxicity in vitro; Ferguson R et al.; Vibrio cholerae neuraminidase (VCN))12.5 units/2 X 10(6) cells/ml) continuously present for a standard 5-day MLC will significant (p less than 0.02) increase the cytotoxic activity generated by a given number of responding spleen cells without reducing the specificity . Heat-inactiviated VCN produced no such augmentation . This augmented cytotoxicity could be reproduced by preincubating (1 hr) the responding spleen cells with VCN (25 units/2.5 X 10(6) cells/ml) before addition of stimulating spleen cells . Preincubating the stimulator spleen cells with VCN had no effect . VCN preincubation of target cells or presensitized effector cells produced no augmentation . The addition of soluble VCN to the killing assay also did not increase cytotoxicity . Thus, VCN acts only during the generation of specifically sensitized cytotoxic T cells . When the effect of VCN on MLC reactivity, cell recovery and total cytotoxicity (lytic units/10(6) cells) were compared, it became apparent that VCN increases the proliferation of responder cells after stimulation resulting in both an increased number of cells and also an increase in the proportion of specifically sensitized cytotoxic cells in the culture . VCN treatment of responder cell membrane apparently permits a more ready response to allogenic antigens in culture facilitating both increased proliferation and the increased development of specific cytotoxic killers.

Med J Aust, 1976 Nov 27, 2(22), 823 - 5
Vibrio parahaemolyticus gastroenteritis associated with international travel; Thomas PM et al.; Food-borne gastroenteritis due to Vibrio parahaemolyticus is becoming of increasing world importance . This paper describes the clinical presentations and laboratory investigations of four cases of gastroenteritis due to V . parahaemolyticus among passengers returning to Australia on international aircraft from South-East Asia.

J Biol Chem, 1976 Nov 25, 251(22), 6929 - 33
Purification and characterization of L-asparaginase with anti-lymphoma activity from Vibrio succinogenes; Distasio JA et al.; Homogeneols L-asparaginase with anti-lymphoma activity was prepared from Vibrio succinogenes, an anaerobic bacterium from the bovine rumen . An overall yield of pure L-asparaginase of 40 to 45% and a specific activity of 200 +/- 2 IU per mg of protein was obtained . The pure enzyme can be stored at -20 degrees for at least 3 months with no loss of activity . The isoelectric point of the L-asparaginase is 8.74 . No carbohydrate, phosphorus, tryptophan, disulfide, or sulfhydryl groups were detected . The enzyme has a molecular weight of 146,000 and a subunit weight of approximately 37,000 . The Km of the enzyme for L-asparagine is 4.78 X 10(-5) M and the pH optimum of the L-asparaginase reaction is 7.3 . D-Asparagine was hydrolyzed at 6.5% of the rate found with the L isomer . L-Glutamine and a variety of other amides were not hydrolyzed at significant rates; the activity of the enzyme for L-glutamine was 130- to 600-fold less than that of other therapeutically effective L-asparaginases of bacterial origin . The L-asparaginase from V . succinogenes is immunologically distinct from the L-asparaginase (EC-2) of Escherichia coli.

J Immunol, 1976 Nov, 117(5 Pt.2), 1774 - 80
Membrane expression of Thy-1,2 and GM1 ganglioside on differentiating T lymphocytes; Milewicz C et al.; C3H mouse bone marrow cells were separated by discontinuous BSA gradient centrifugation . Marrow cells from the 17 to 19%, 19 to 21% to 23%, 23 to 25%, 25 to 27% interfaces and the cell pellet were treated with thymic factor (TF) or with Vibrio cholerae neuraminidase (VCN), followed by anti-Thy-1,2 and anti-GM1 ganglioside antisera . Antigens determined by anti-Thy-1.2 or anti-GM1 were expressed either with TF or VCN within a 30-min incubation . Cells expressing these antigens after VCN or TF treatment were concentrated in the 19 to 21% layer and the 21 to 23% layer whereas there was little or no antiserum cytotoxicity in the other layers . A small amount of Thy-1.2 of GM1 could be detected by cytotoxicity on 19 to 21% layer marrow cells within 15 min of either TF or VCN treatment . Treatment of 19 to 21% layer cells with TF or VCN had no effect on anti-H-2 cytotoxicity . Pretreatment of TF-treated marrow cells with cholera toxin or choleragenoid (which bind cell surface GM1) abrogated the cytotoxicity of anti-Thy-1.2 or anti-GM1 without affecting anti-H-2 cytotoxicity . Pretreatment of Thy-1.2 positive C3H thymocytes with cholera toxin or choleragenoid greatly reduced the cytotoxicity of anti-Thy-1.2 antisera without affecting the cytotoxicity of anti-H-2 antisera . Nude mouse splenocytes, after treatment with VCN or TF, were susceptible to the lytic action of anti-Thy-1.2 and anti-GM1 . The possibility that nonspecific autologous antibodies were responsible for anti-Thy-1.2 cytotoxicity toward VCN-treated marrow cells was eliminated because anti-Thy-1.2 was not cytotoxic for VCN treated AKR (Thy-1.1) marrow cells.

Am J Med Sci, 1976 Nov-Dec, 272(3), 331 - 4
Vibrio fetus endocarditis in a patient with systemic lupus erythematosus; Dzau VJ et al.; Vibrio fetus endocarditis occurred in a patient with systemic lupus erythematosus receiving azathoprin and prednisone . Blood cultures required 14 days to become positive . The fastidious growth requirement of this organism is reviewed because lack of appreciation of these may result in failure to make the diagnosis . This is the first reported case of Vibrio fetus endocarditis occurring in the setting of a connective tissue disorder and immunosuppressive therapy.

Zh Mikrobiol Epidemiol Immunobiol, 1976 Nov, (11), 84 - 7
{Experimentally induced cholera in guinea pigs . I . Elaboration of the method of infection}; Kotenok IaF et al.; The method of intrapulmonary infection of guinea pigs was suggested for the assessment of the virulent properties of cholera vibrios . Addition into the diluent of 10% peptone, 10% gelatine and 0.05% agar-agar led to the reduction of LD50 by over 1000 times . A specific infectious process coursing in an acute generalized form with bacteriemia and affection of the small intestine developed in the infected animals . The majority of the animals perished in 1 to 2 days.

Zh Mikrobiol Epidemiol Immunobiol, 1976 Nov, (11), 55 - 8
{A comparative study of ultrastructural organization of the nonagglutinating vibrios and the causative agents of cholera}; Shul'ga MA; A common plan of ultrastructural organization was revealed in a comparative study of the ultrastructure of the nonagglutinating El Tor and cholera vibrios: the presence of a three-layer cell wall and cytoplasmic membrane, the presence of vesicles on the external layer of the cell wall or between the cell wall and the cytoplasmic membrane; identity of the intracytoplasmic membranous structures; the presence of intracytoplasmic inclusions in the form of granules of various size and electron density; cell division by means of a band with the formation of two cells of equal size or of many bands in one cell, and also by the joining of the cytoplasmic membrane with the formation of rings at the sites of cell division . Identify of the ultrastructure of the vibrios under study can serve as confirmation of the views of the investigators who consider that nonagglutinating vibrios should be referred to a single V . cholerae species.

Zh Mikrobiol Epidemiol Immunobiol, 1976 Nov, (11), 115 - 9
{A study of immunological efficacy of the oral chemical cholera vaccine on experimental animals}; Dzhaparidze MN et al.; Three fractions isolated from the culture fluid of the 569B cholera vibrio strain were studied . The fractions differed by the extent of purification, and the content of the toxid and the O-antigen . In intraintestinal application to rabbits all of them caused formation of antitoxins and vibriocidal antibodies in the blood serum . The immunizing dose of the preparations in intraintestinal administration exceeded the dose required for subcutaneous application . Fraction I should be used for Producing tablet form of cholera vaccine because it was least toxic and provided the greatest yield.

Zh Mikrobiol Epidemiol Immunobiol, 1976 Nov, (11), 108 - 10
{The protective and immunodepressive activity of cell fractions of a cholera-like vibrio}; Stanislavskii ES et al.; A study was made of the protective and immunodepressive activity of the sytoplasmic fractions of a cholera-like vibrio . Ribosomal fraction proved to possess more marked protective and immunodepressive properties than the soluble cytoplasmic fraction.

J Clin Pathol, 1976 Nov, 29(11), 1014 - 5
Marine vibrios associated with superficial septic lesions; Ryan WJ; Three cases are reported in which a marine vibrio, Vibrio alginolyticus, was isolated from superficial septic lesions . All cases had been exposed to sea-water . The possible significane of these findings and the need for further investigations are discussed.

Appl Environ Microbiol, 1976 Nov, 32(5), 679 - 84
Membrane filter procedure for enumeration of Vibrio parahaemolyticus; Watkins WD et al.; A membrane filtration procedure has been developed for the enumeration of Vibrio parahaemolyticus in marine waters . Background microbial growth on the primary medium was decreased through the use of sodium cholate and copper sulfate, high pH, 3% NaCl, and an elevated incubation temperature . A series of in situ tests was employed to obviate the picking of colonies for identification; thereby, the enumeration of V . parahaemolyticus was accomplished within 30 h . Confirmation of typical colonies approached 95% . Relative to immediate plating on brain heart infusion agar spread plates, the recovery of V . parahaemolyticus cells suspended in phosphate-buffered saline or in seawater held for 24 h at 4 to 6 degrees C was about 90% . Assay variability did not exceed that expected by chance . Recoveries of V . parahaemolyticus from coastal and estuarine surface waters exceeded those obtainable by other methods examined.

Zentralbl Bakteriol {Orig A}, 1976 Nov, 236(2-3), 288 - 93
Effect of cholera enterotoxin preparations on cutaneous response in rabbit under varied conditions; Singh A et al.; Enterotoxic activity of two preparations obtained from Vibrio cholerae, B-53-6 Inaba and B-53-10 Ogawa was tested in ligated ileal loops of rabbit . The biologically active enterotoxic preparations were further used to study the permeability reaction in rabbit skin . Cutaneous response tended to be linear only with higher concentrations of the toxin and showed maximum blueing intensity between 16 and 24 hrs of intracutaneous inoculation . Exposure of enterotoxin preparations to elevated temperatures greatly reduced the cutaneous response and the activity was completely lost at 100 degrees C . Change in pH towards alkaline side lowered the permeability activity to lesser extent as compared to a shift on acidic side . However, a residual activity could still be detected when pH of the enterotoxins was lowered to 3 at 30 degrees C for 4 hrs.

Infect Immun, 1976 Oct, 14(4), 1028 - 33
Isolation of a factor causing morphological changes of chinese hamster ovary cells from the culture filtrate of Vibrio parahaemolyticus; Honda T et al.; A factor changing Chinese hamster ovary cells from an oval to a spindle shape was isolated from the culture filtrate of Vibrio parahaemolyticus . It was partially purified by successive column chromatographies on diethylaminoethylcellulose, diethylaminoethyl-Sephadex A-25, hydroxylapatite, and Sephadex G-200 . This factor was separated from thermostabl direct hemolysin and was heat labile.

Appl Environ Microbiol, 1976 Oct, 32(4), 617 - 22
Morphological characterization of small cells resulting from nutrient starvation of a psychrophilic marine vibrio; Novitsky JA et al.; Upon starvation, Ant-300, a psychrophilic marine vibrio, was observed to decrease in size and change in shape from a rod to a coccus . After 3 weeks of starvation 50% of the starved population was able to pass through a filter with a pore size of 0.4 mum . Electron microscopy of thin sections of the small cells revealed normal cell structure except for an enlarged periplasmic space . When inoculated into a fresh medium, starved cells growth without a significant lag and regained "normal" size and shape within 48 h.

Can J Microbiol, 1976 Oct, 22(10), 1437 - 42
Extracellular nuclease produced by a marine bacterium . I . Extracellular deoxyribonuclease formation by a marine Vibrio sp; Maeda M et al.; Deoxyribonuclease (DNase) activity was found in the culture fluids of numerous marine bacteria isolated from seawater . Among these ogranisms, marine bacterium, Vibrio sp., strain No . 2, showed the highest deoxyribonucleic acid-hydrolyzing activity . This organism requires salts of seawater for both growth and extracellular DNase formation . The DNase activity could not be detected in the synthetic seawater culture liquid lacking magnesium ion, and DNase activity decreased in a calcium-deficient medium . The optimum temperature for the growth of this organism was between 15 and 25 degrees C . The formation of extracellular DNase was the greatest at 20 degrees C and less activity was found at 10 and 30 degrees C.

Cancer Res, 1976 Oct, 36(10), 3561 - 7
Bacterial proteinaceous products (bacteriocins) as cytotoxic agents of neoplasia; Farkas-Himsley H et al.; Several bacteriocins, bacterial proteinaceous antibiotics, are shown to markedly inhibit the division of various established (neoplastic) mammalian cell lines . The bacteriocins tested originated from Escherichia coli, Pseudomonas aeruginosa, Vibrio cholerae, and Vibrio eltor . Using exponentially growing L60T mouse fibroblasts, the inhibitory effect was concentration dependent, and a growth inhibitory unit, equivalent to cytotoxic index 50, was established . Expression of toxicity as a function of duration of exposure to pyocin required 3 to 4 hr . DNA synthesis was inhibited and reflected the effects on growth inhibition . Maximal sensitivity to the bacteriocin was observed prior to mitosis in the G2 phase of the cell cycle.

Can J Comp Med, 1976 Oct, 40(4), 392 - 6
A direct fluorescent antibody test for large spirochetes in swine dysentery using hyperimmunized swine serum; Lee CH et al.; A direct fluorescent antibody test was developed for the identification of large spirochetes which are considered to be the cause of swine dysentery . Sera from swine which had recovered from swine dysentery and had been hyperimmunized by the intravenous and intraperitoneal injection of filtered spirochetes were used for conjugation with fluorescein isothiocyanate . A bright greenish fluorescence of large spirochetes was observed with the conjugated serum from hyperimmunized pig No . 1 when diluted 1:8 and hyperimmunized pig No . 2 when diluted 1:2 . Pig No . 1 had developed a serum titer of 1:64 using the indirect fluorescent antibody test for large spirochetes . The conjugated serum from the three swine which had recovered from swine dysentery fluoresced spirochetes only when undiluted . The conjugated serum from the two swine treated while having a hemorrhagic diarrhea did not fluoresce spirochetes . No immunofluorescence of Vibrio spp . was observed.

Aust J Exp Biol Med Sci, 1976 Oct, 53(5), 361 - 72
Vibrio cholerae infection and immunity in mice; Lee MM et al.; Lyophilized cultures of V . cholerae 569B slowly lose their virulence for neonatal and adult mice during long term storage . Following a single passage in orally infected 6-day old mice, a highly virulent strain (designated 569B/MP) was isolated . This organism causes rapidly fatal intestinal infections in 6-day old mice; large numbers can be isolated in pure culture from the intestinal fluid . Freezing and storage at -60 degrees of dead animal provides a simple means of maintaining the high virulence of the culture over a period of at least 9 months . This strain produces choleracic symptoms and death in approx . 50% of adult mice following oral infection . Large numbers of viable organisms may also be isolated from the small intestine over a period of at least 40 h . These criteria have been used as a basis for assessing protection against cholera infection induced by immunization with living or heat-killed V . cholerae given orally or intraperitoneally.

Can J Microbiol, 1976 Oct, 22(10), 1443 - 52
Extracellular nuclease produced by a marine bacterium . II . Purification and properties of extracellular nuclease from a marine Vibrio sp; Maeda M et al.; Extracellular nuclease produced by a marine Vibrio sp., strain No . 2, was purified by salting out with ammonium sulfate and by chromatography on a DEAE-cellulose column and twice on a Sephadex G-200 column . The nuclease was eluted as a single peak in which the deoxyribonuclease (DNase) activity and ribonuclease (RNase) activity appeared together . Polyacrylamide disc gel electrophoresis showed a single band of stained protein which had both DNase and RNase activity . The molecular weight of the enzyme was estimated to be 100 000 daltons . When using partially purified enzyme from the DEAE-cellulose column, the optimum pH for activity was 8.0, and the enzyme was activated strongly by 0.05 M Mg2+ ions and stabilized by 0.01 M Ca2+ ion . These concentrations of Mg2+ and Ca2+ ions are similar to those of the two cations in seawater . Indeed, the enzyme revealed high activity and strong stability when kept in seawater . The presence of particulate matter, such as cellulose powder, chitin powder . Hyflosupercel, Kaolin, and marine mud increased the stability of the enzyme . When the hydrostatic pressure was increased from 1 to 1000 atmospheres, the decrements of the enzyme activity were more pronounced at 30 and 40 degrees C than at 25 or 50 degrees C . The enzyme activity was restored after decompression to 1 atm at 30 degrees C.

Biochemistry, 1976 Sep 21, 15(19), 4228 - 33
Maleylacetone cis-trans-isomerase: affinity chromatography on glutathione-bound sepharose . Two-substrate-binding sequence from inhibition patterns; Morrison WS et al.; Maleylacetone cis-trans-isomerase isolated from Vibrio 01 binds glutathione strongly; Km = 1.4 X 10(-4) M . Oxidized glutathione and S-methylglutathione are competitive inhibitors, KI = 9.4 X 10(-4) and 1.2 X 10(-3) M, respectively . Based on these interactions, three different glutathione-bound agarose affinity adsorbents were synthesized and tested . Affinity chromatography of the isomerase with one of these affords 70- to 100-fold purifications . In separate syntheses, portions of the affinity arm were prepared and examined as to their inhibitory properties in the enzyme-catalyzed reaction . The fragment, containing glutathione bound through its sulfur to the carbon chain, is a powerful competitive inhibitor for glutathione (KI = 6 X 10(-5) M) . The results described suggest that the isomerase binds glutathione through the backbone of the tripeptide and that the thiol group is required for activity . The initial velocity patterns of the enzyme-catalyzed reaction resulting from simultaneous variation of glutathione and maleylacetone concentrations were examined in the absence and presence of inhibitors resembling glutathione . The observed kinetic patterns suggest an ordered sequence of binding maleylacetone first followed by glutathione.

Eur J Clin Invest, 1976 Sep 10, 6(5), 347 - 53
Studies on the mitogenic activity of trypsin, pronase and neuraminidase on human peripheral blood lymphocytes; Girard JP et al.; Human peripheral blood lymphocytes were cultured in the presence of protease (trypsin and pronase) and Vibrio choleraneuraminidase . T and B lymphocyte populations were separated and the effect of these enzymes plus phytohaemagglutin or Tuberculin was studied . The results of these experiments show that proteases moderately stimulate spontaneous deoxynucleic acid synthesis of control cells and potentiate the effect of tuberculin on sensitized cells . These enzymes act specifically on B lymphocytes . Neuraminidase also increases spontaneous deoxyribonucleic acid synthesis of control cells and augments significantly the in vitro response to PPD . There is no additive effect of neuraminidase on phytohaemagglutin stimulated cells . Neuraminidase seems to stimulate specifically T lymphocytes . Some possible mechanisms of action of these enzymes are proposed and discussed.

Can Med Assoc J, 1976 Sep 4, 115(5), 397 - 8
Laboratory investigation and infection control of cholera in Kingston, Ont; Lewis RG et al.; Vibrio cholerae, biotype El Tor, was isolated in a hospital laboratory in Kingston, Ont . in 1974 . Confirmation and complete identification by the Ontario regional and provincial public health laboratories was obtained within 3 days . Institution of well established infection-control and public health measures prevented spread of the infection within the hospital and the community.

Can Med Assoc J, 1976 Sep 4, 115(5), 393 - 6
A case of cholera in Kingston, Ont; Bourdages R et al.; A case of cholera occurred in Kingston, Ont . in 1974 in a traveller from South Africa . Treatment, based on an understanding of the pathophysiology of cholera diarrhea and the mechanism of action of the Vibrio cholerae enterotoxin on gastrointestinal fluid loss, consisted of correcting the severe loss of fluid and electrolytes and the metabolic acidosis, as soon as the patient could tolerate taking fluids orally, further fluid replacement consisted increasingly of oral administration of glucose and saline . Tetracycline therapy was given only to shorten the duration of the acute illness.

Southeast Asian J Trop Med Public Health, 1976 Sep, 7(3), 377 - 9
Hydrogen sulphide production as an aid to the identification of Vibrio parahaemolyticus; Jegathesan M et al.; In this study 18 strains of Vibrio parahaemolyticus from food and 8 from humans were tested for hydrogen sulphide production on various modifications of Russel's Triple Sugar slopes and on TSI . All strains showed a characteristic surface browning on RTS with Andrade's indicator . This was not seen when RTS with phenol red as indicator or TSI were used . Appearance of this phenomenon allows unknown strains to be suspected as being Vibrio parahaemolyticus.

Zh Mikrobiol Epidemiol Immunobiol, 1976 Sep, (9), 70 - 4
{Avidity of specific antibodies in the sera of cholera patients and vibrio carriers}; Bunin KV et al.; Avidity of specific antibodies was studied in the sera of patients suffering from cholera and vibrio-carriers by the rate of antibody binding (in the bacterial agglutination reaction) and by the completeness of their binding with O-antigen (in the passive hemagglutination test) . There were revealed statistically significant higher indices of antibody avidity contained in the blood sera of the vibrio carriers, in comparison with the antibodies in the blood sera of patients with clinically manifest forms of cholera . There proved to be an increase in the antibody avidity in the dynamics of the infectious process . It is supposed that the degree of avidity of anticholera antibodies in examination of whole sera depended on the ratio of the specific macro- and microglobulins in them.

J Natl Cancer Inst, 1976 Sep, 57(3), 717 - 20
Immunotherapy and chemotherapy in children with neuroblastoma; Nesbit ME et al.; Recent advances with immunotherapy in animal tumors suggested that trials with a combination of chemotherapy and immunotherapy in human malignant tumors might be worthwhile . A pilot program with Vibrio cholera neuraminidase-treated tumor cells plus BCG was tested in 3 patients who had had chemotherapy for disseminated neuroblastoma . Two of these children were in "complete remission" after radiation therapy and chemotherapy before the administration of immunotherapy . Relapse occurred in 5-6 months in all 3 patients . These disappointing results are discussed in relation to problems of current chemotherapy in disseminated neuroblastoma including results obtained at second-look operations in patients obtaining "complete remission."

Can J Microbiol, 1976 Sep, 22(9), 1263 - 8
Sparing effect of lithium ion on the specific requirement for sodium ion for growth of Vibrio parahaemolyticus; Morishita H et al.; The role of NaCl in the growth of Vibrio parahaemolyticus strain A-55 was investigated . Maximal growth was obtained at 0.5 M NaCl in a synthetic medium . Na+ could not be replaced for growth by any other cations or substances . When the medium was kept isotonic with 0.5 M NaCl by the addition of sucrose, good growth was obtained with 0.1 M NaCl . By reducing the osmotic pressure and decreasing the NaCl concentration from 0.5 M to 0.1 M, the same growth yield was obtained as when using a medium containing 0.02 M NaCl and 0.08 M LiCl . This was not the case with sucrose . Therefore, it is concluded that ionic strength and osmotic pressure are important environmental factors affecting growth . The minimal essential requirement for Na+ for growth of V . parahaemolyticus was 0.003 M, because this was never replaced with any other cations and agents . Hence, requirement for Na+ for growth involves a specific requirement for Na+, ionic strength, and osmotic pressure, respective concentrations being 0.003 M, 0.047 M, and 0.45 M . Osmotic support was required for growth when the concentration of NaCl was decreased to 0.05 M . The effect of ionic strength of monovalent cations other than Na+ on growth was examined at 0.003 M NaCl . Among LiCl, NH4Cl, KCl, and RbCl, Li+ was the most accelerative for growth in the synthetic medium.

Infect Immun, 1976 Sep, 14(3), 687 - 93
Development of a purified cholera toxoid . III . Refinements in purification of toxin and methods for the determination of residual somatic antigen; Rappaport RS et al.; The addition of an ultrafiltration step to the purification procedures previously described for cholera toxin (Rappaport et al., (1974) permitted the preparation of highly purified antigenic toxoids essentially free of somatic antigen(s) . The purity of such toxoids is established: (i) by the absence of more than about one part limulus amebocyte lysate (LAL)-positive endotoxin per 10(5) parts toxoid and (ii) by the inability of the toxoids to elicit a significant rise in rabbit vibriocidal antibody . The antigenicity of the toxoids is demonstrated by their ability to produce the same high levels of rabbit serum antitoxin as are produced by comparable toxoids containing small amounts of somatic antigen . The results also indicate that amounts of somatic antigen of the order of less than or equal to 1 mug/100 mug of toxoid do not exert an adjuvant effect on the toxoid, at least with respect to circulating antitoxin . Other data show that, where present, the ability of somatic antigen to elicit vibriocidal antibody is influenced by the immunization schedule employed and that a correlation exists between the LAL-determined endotoxin content of the toxoids and their ability to stimulate vibriocidal antibody . Somatic antigen-free toxoids, purified and tested by the refinements herein described, were prepared for use in the National Institutes of Health sponsored field trials, and data pertaining to their purity and antigenic properties are presented.

Cancer Res, 1976 Sep, 36(9 pt.1), 3051 - 7
Correlations between humoral immunity and successful chemotherapy-immunotherapy; Cantrell JL et al.; Experiments were designed to evaluate the characteristics of the humoral immune response induced by active immunotherapy, both specific (neuraminidase-treated tumore cells) and nonspecific (Bacillus Calmette-Guerin organisms), in the L1210-C57BL/6 X DBA/2F tumor-host system . Tumor burden was minimized with chemotherapy (1,3-bis-(2-chloroethyl)-1-nitrosourea) prior to immunotherapy . A marked increase in the concentration of serum immunoglobulins (immunoglobulin M, immunoglobulin G, and immunoglobulin G) was observed following successful therapy . The highest concentration of these immunoglobulins was found in mice given both Vibrio cholerae neuraminidase-treated cells and B . Calmette-Guerin after chemotheraphy . Tumor-specific immunoglobulin M and immunoglobulin G, as measured by indirect immunofluorescnece, were detected in the sera during the course of successful therapy . Positive immunofluorescence was not observed with progressive sera . Complement-dependent cytotoxic activity against L1210 cells was first detected 5 days after immunotherapy, and increased for several weeks . A high level of cytotoxic activity correlated with successful therapy, whereas low levels were foun in treated mice with recurring tumors . Serum cytotoxicity was not detected in untreated mice with progressively growing tumor.

Mol Gen Genet, 1976 Aug 2, 146(3), 275 - 83
Stability of "spacer" sequences of pre-ribosomal RNA in Escherichia coli; Kano Y et al.; "SPACER" SEQUENCES OF AN RRNA gene transcript were detected with high efficiency by hybridization with DNA of the specilized transducing phase phi80rrn . Hybridization-competition studies revealed that 20 to 23% of the 30S precursor rRNA, obtained from E . coli mutant strain AB301/105, consist of "spacer" sequences . The "spacer" sequences formed hybrids with E . coli DNA, but not with Vibrio DNA . Experiments with RNA labeling in the presence of rifampicin showed that more than 80% of the spacer sequences arrive in full-length 30S pre rRNA chains before any cleavage of the RNA occurs . The hybridization assays also permitted the detection of "spacer" sequences in pulse-labeled rRNA of wild-type cells, in which the 30S pre-rRNA is already cleaved during its synthesis . Many of these "spacer" sequences degraded to alcohol-soluble materials with a half-life time of 1.2 min . The half-life was not lengthened by the treatment of cells with chloramphenicol, which stabilizes bulk mRNA . However, unstable "spacer" sequences transcribed in cells deficient in RNase III exhibited slower degradation, with a half-life time of about 9 min, whereas the cleavage of 30S pre-rRNA to smaller RNA species occurred with a half-life of about 3 min . These results are consistent with the notion that a rate-limiting action of RNase III in the initial attack leads to degradation of "spacer" sequences in rRNA gene transcript; and that degradation is not at all connected with ribosome translocation.

Biull Eksp Biol Med, 1976 Aug, 82(8), 961 - 2
{Study of the heterogenetic antigens in vaccinal preparations of V . cholerae}; Zhukov-Verezhnikov NN et al.; There proved to be a serological similarity between the antigens of the human small intestine, the stomach and the liver, and the antigens of various cholera vibrio fractions . No antigenic similarity was revealed in examination of the heart and kidney . Heterogenous antigen was found not only in the somatic V . cholerae antigen, strain 569 (B), but also in the cholerogen-toxoid obtained from it . At present it is the most widespread prophylactic preparation.

Appl Environ Microbiol, 1976 Aug, 32(2), 288 - 93
Biochemistry of Vibrio cholerae virulence: purification of cholera enterotoxin by preparative disc electrophoresis; Lewis AC et al.; Procedures for cholera enterotoxin purification previously developed in this labarotory were not applicable to large-scale purification, and these methods resulted in low yields of pure toxin . An efficient scheme has been developed whereby pure cholera enterotoxin can be obtained from 6 to 8 liters of culture supernatant fluid . This method consists of concentration by membrane ultrafiltration followed by gel filtration and cation-exchange chromatography . Pure cholera enterotoxin of high biological potency was obtained after a final step of preparative acrylamide gel electrophoresis . The degree of purity of the toxin-antigen as well as its biological activity were determined at various setps of purification . This alternate technique for purification is offered because of the widespread interest in cholera enterotoxin as a specific stimulator of adenyl cyclase.

Infect Immun, 1976 Aug, 14(2), 527 - 47
Vibrio cholerae adherence and colonization in experimental cholera: electron microscopic studies; Nelson ET et al.; Colonization of the intestinal epithelium by Vibrio cholerae was examined in two model systems, in ligated ileal loops of adult rabbits and in the patent gut of infant rabbits, using both scanning and transmission electron microscopy . Time studies in the adult model showed a lag period of up to 1 h before the attachment of significant numbers of the vibrios . The bacteria appeared initially in small patches on the sides of the villi, predominantly along the transverse furrows . The number of adherent bacteria steadily increased, reaching a maximum between 4 and 7 h, when a dense mat of bacteria several layers thick covered much of the villi . After this time there was a rapid decline in the number of V . cholerae bound . By 12 to 16 h only a few bacteria could be seen on the surface of the villi, which had a rough, patchy appearance at these later times . Globular protrusions, with vibrios attached, may play a role in the clearance of bacteria . Colonization and clearance in the patent intestine of the infant rabbit occurred much as in the adult model . However, the bacteria adhered more uniformly and there was no lag in attachment . In both models the majority of bacteria were aligned horizontally with the epithelial surface, but some were attached in an end-on manner, with their flagella extending into the lumen . The bacteria adhered via their surface coats directly to the tips of the microvilli, except for a few vibrios that were partly embedded into the brush border . Some changes in the microvilli occurred as a consequence of the bacterial attachment.

Infect Immun, 1976 Aug, 14(2), 363 - 7
Cell-mediated immunity to Vibrio cholerae with ribonucleic acid-protein fractions of V . cholerae L-form lysates; Agarwal SC et al.; Different L-form lysate vaccines of Vibrio cholerae serotypes Ogawa and Inaba and their combination along with ethyl alcohol-precipitated ribonucleic acid (E-RNA) and phenol-extracted RNA (P-RNA) fractions of V . cholerae Ogawa lysates were tested for production of cell-mediated immunity . Both E-RNA and P-RNA fractions induced an increase in leukocyte migration inhibition, macrophage migration inhibition, and macrophage aggregation . They also induced delayed hypersensitivity in rabbits . More consistent results were obtained with the P-RNA fraction.

J Clin Microbiol, 1976 Aug, 4(2), 133 - 6
Enhancement by sodium chloride of the selectivity of thiosulfate citrate bile salts sucrose agar for isolating Vibrio cholerae biotype El Tor; Morris GK et al.; In this study we utilized the salt-tolerant characteristics of vibrios to develop a more selective medium by addition of NaCl to thiosulfate citrate bile salts sucrose (TCBS) agar . The effect of adding salt to TCBS agar varied greatly among brands of TCBS agar and between lots of the same brand . The addition of salt at concentrations as high as 1.5% (2.5% total NaCl) caused the inhibition of growth of three species of commonly encountered normal bowel flora and one strain of classical Vibrio cholerae but did not compromise the use of TCBS agar for isolation of V . cholerae biotype El Tor.

Can J Microbiol, 1976 Aug, 22(8), 1186 - 7
Photoreactivating property of the Vibrio cholerae cell system; Chanda PK et al.; Ultraviolet-inactivated chlora phage PL163/10 underwent a maximum of abot 36% of photoreactivation within the host V . cholerae cells . UV-inactivated but non-infected V . cholerae cells also underwent the same degree of photoreactivation.

Am J Dig Dis, 1976 Aug, 21(8), 613 - 7
A bioptic study of gastrointestinal mucosa in cholera patients during an epidemic in southern Italy; Pastore G et al.; A histological biopsy study of gastric and jejunal mucosa of eight acute cholera patients during an epidemic in Southern Italy was carried out . The study demonstrated in all patients an intact epithelial lining of gastric and jejunal mucosa, a moderate degenerative process of enterocytes, presence of inflammatory lesions manifested by edema, vascular congestion, mononuclear cell infiltrate of lamina propria, and discharge of goblet-cells mucus . These changes reverted to normal in a few days . The authors emphasize that, contrary to cholera patients of Asiatic areas in whom an underlying chronic spruelike enteropathy is very common, the histological picture observed in Western patients may be considered more specific since Vibrio cholerae acts upon a normal intestinal mucosa.

Histochemistry, 1976 Jul 30, 48(1), 71 - 80
Cytochemistry of colloidal iron binding to the surface of Hela cells and human erythrocytes; Mareel M et al.; It seems from the literature that colloidal iron (C.I.) binding sites on cell surfaces cannot be completely removed by treatment with Vibrio Colerae alpha-neuraminidase . We wondered if C.I . particles bind to negative groups other than the carboxyl groups of sialic acids . Using HeLa cells from suspension cultures and fresh human erythrocytes, we examined, with the transmission electronmicroscope, the influence of the following enzymatic and histochemical treatments on C.I . staining: alpha-neuraminidase; hyaluronidase; ribonuclease; alpha-amylase; mild methylation (MM); MM + saponification (Sap.); MM + Sap +MM; MM + Sap + alpha-neuraminidase; active methylation (AM); AM + Sap; AM + Sap + AM; AM + Sap + alpha-neuraminiadase; CH3OH (80%); Sap . It seemed from these experiments that the carboxyl groups of alpha-neuraminidase sensitive sialic acids constitute the majority of binding sites for C.I . to these particular cells . The most interesting candidates for the residual binding of C.I . are carboxyl groups of alpha-neuraminidase resistant molecules, sulfon, sulfin, and sulfate groups.

Biull Eksp Biol Med, 1976 Jul, 82(7), 841 - 3
{NAG infection of grass frog (Rana temporaria) tadpoles}; Avtsyn AP et al.; Rana temporaria tadpoles (2250) were contaminated with three types of NAG vibrio cultures . Clinical, bacteriological and morphological examinations showed the larvae to be suffering from an acute infection during the first 2 days after the contamination . Then the vibrios persisted in the tadpole organism for a long time and were excreted in the feces.

Zh Mikrobiol Epidemiol Immunobiol, 1976 Jul, (7), 46 - 8
{The spread of parahemolytic vibrios in humans in the Burgass Region (Bulgaria)}; Zakhariev ZA; The presence of parahemolytic vibrios in the water of the Black sea and also in other water bodies, in the hydrobionts, and later in the secondarily contaminated meat products was demonstrated in 1968 . In 1973 gastroenteritis caused by parahemolytic vibrios became widespread; 126 cases were confirmed bacteriologically, and 3 cases were found to be carriers . The infected food products were eliminated, and no more cases of the disease were revealed in 1974.

Zh Mikrobiol Epidemiol Immunobiol, 1976 Jul, (7), 131 - 4
{Clinical cases of food poisoning caused by NAG-vibrios}; Medinskii GM et al.; On the basis of bacteriological and serological studies carried out during two outbreaks of food poisoning it was found that the disease was caused by NAG-vibrios of the I Heiberg group, serotypes 5 and 34 after Sakazaki . The disease was accompanied by isolation of NAG-vibrios from feces and the washings from the stomach, it was characterized by symptoms of acute gastroenteritis and enteritis of mild and moderate severity . The incubation period lasted from 3 to 30 hours . There was an increase in the agglutinin titres to homologous strains of NAG-vibrios . Of the 6 strains tested 4 proved to be enteropathogenic in intraintestinal infection of rabbits in doses of 10(5) microbial cells.

Infect Immun, 1976 Jul, 14(1), 240 - 5
Adhesive properties of Vibrio cholerae: nature of the interaction with isolated rabbit brush border membranes and human erythrocytes; Jones GW et al.; Nonmotile vibrio mutants lacked the ability to adhere to rabbit intestinal brush border membranes and to agglutinate human group O erythrocytes, but motile revertant vibrios isolated from such strains expressed adhesiveness equivalent to that of the original parent . Two possible explanations for the relation between vibrio motility and adhesion in these assays systems are (i) that the rate of adhesion depends on the rate of chance contact brought about by motility, and (ii) that the flagellum either acts as a carrier for the bacterial adhesin or may itself be the adhesin . The present study indicates, however, that the lack of adhesion by nonmotile vibrios did not depend on motility as such and did not involve greater rates of elution . Increasing the rate of contact between nonmotile vibrio mutants and brush border membranes by compaction did not restore the adhesive properties of the defective strains . Accordingly, we speculate that the flagellum may function in some indirect way that allows the expression of the adhesive properties, such as by acting as a carrier for a specific vibrio adhesin . Adhesion to brush borders and agglutination of human group O erythrocytes was specifically inhibited by L-fucose and various glycosides of L-fucose and to a lesser extent by D-mannose . Vibrios adhered specifically to agarose beads that carried covalently linked L-fucose on their surfaces . The results suggest that L-fucose-containing structures of eukaryotic cell surfaces may function as receptors for the vibrio adhesin and may therefore be an important determinant of host susceptibility.

Infect Immun, 1976 Jul, 14(1), 232 - 39
Adhesive properties of Vibrio cholerae: adhesion to isolated rabbit brush border membranes and hemagglutinating activity; Jones GW et al.; Adhesion of vibrios to the small intestine may occur (i) by association of the bacteria with secreted mucus gel or (ii) by adherence of the bacteria to the surface of epithelial cells . In the present study, vibrios readily adhered to isolated brush border membranes obtained from rabbit intestinal epithelial cells . Adhesion was temperature dependent and required the presence of divalent cations such as calcium . The agglutination of human O erythrocytes by Vibrio cholerae was observed also, and the hemagglutination test appeared to detect the same mechanism that was involved in the adhesion of vibrios to brush borders . When the bacteria were grown in broth they were adhesive and hemagglutinating, but vibrios grown on agar plates or suspended in buffer for 15 min at 37 C lacked these abilities, even though they retained undiminished motility . These two model systems differed, however, in that strontium promoted only adhesion to brush borders . The significance of this difference remains to be determined . Vibrios were observed to penetrate intestinal mucus gel and occasionally to become entrapped in it . However, there was no evidence that vibrios attached to mucus gel.

Infect Immun, 1976 Jul, 14(1), 1 - 5
Inactivation of the biological activities of the thermostable direct hemolysin of Vibrio parahaemolyticus by ganglioside Gt1; Takeda Y et al.; Biological activities of the thermostable direct hemolysin produced by Vibrio parahaemolyticus, such as its hemolytic activity and lethal activity, were inhibited by neuraminidase-sensitive gangliosides, of which GT1 ganglioside was the most inhibitory . Neuraminidase-resistant gangliosides did not affect the activities of the hemolysin . Results showed that horse erythrocytes, which are resistant to the hemolysin, do not contain the neuraminidase-sensitive gangliosides GT1 and GD1a . Therefore, we propose that neuraminidase-sensitive gangliosides, and especially GT1 ganglioside, may be the receptor sites on the membranes for the thermostable direct hemolysin of V . parahaemolyticus.

J Clin Invest, 1976 Jul, 58(1), 91 - 6
Intestinal myoelectric activity in response to live Vibrio cholerae and cholera enterotoxin; Mathias JR et al.; The myoelectric response of the rabbit ileum was studied in response to live Vibrio cholerae culture, a whole cell lysate of cholera, and the purified enterotoxin . Each cholera preparation produced a series of highly organized migrating action potential complexes (MAPC) . An MAPC was defined as action potential discharge with a duration of 2.5 s or longer, followed by similar activity on at least one other consecutive electrode site . The mean and modal onset time of MAPC activity occurred 4 h after the infection with live Vibrio cholerae culture, the freeze-dried whole cell lysate preparation, or the purified enterotoxin . After the onset of activity this pattern persisted for the duration of the recording period (up to 12 h) . The MAPC had a mean propagation velocity of 0.85+/-0.07 cm/s (mean+/-SEM), which remained constant with time . Direct visual observation of the loop revealed that the MAPC's resulted in contractions that propelled intraluminal contents in an aborad direction . The mean fluid output from the 12-cm ileal loops was 6.4+/-1.1 ml/h (mean+/-SEM) . Control experiments consisted of recordings from: (a) a ligated ileal loop into which nothing was placed; (b) a ligated ileal loop into which either uninfected culture broth or 0.9% NaCl solution was injected; (c) a ligated ileal loop infused with 0.9% NaCl solution at a rate of 11.2 ml/h, and (d) rapid injection of 1.0, 2.5, 5.0, or 10.0-ml boluses of 0.9% NaCl into the proximal catheter . MAPC activity was not observed in any of the control experiments . These studies indicate that in addition to a secretory component to cholera, there exists a highly organized MAPC that results in contractions that propel intraluminal contents in an aborad direction.

Health Lab Sci, 1976 Jul, 13(3), 197 - 202
Demonstration of agglutinating antibodies against Vibrio parahaemolyticus in the yellow tail flounder (Limanda ferruginea); Ortiz JS et al.; Sera from the salt water fish (Yellow tail flounder) and the fresh water fish (rainbow trout) were tested for the presence of circulating antibodies against nine strains of Vibrio parahaemolyticus and one of V . alginolyticus . Thirteen out of 26 flounder sera tested gave agglutinating reactions against one specific strain of V . parahaemolyticus; strain No . 3525 . Seven rainbow trout sera tested failed to agglutinate any of the strains tested . All agglutinating activity was lost upon boiling the antigen, therefore, the fish antibodies are due to surface antigens (K-antigens) and not to somatic antigens . The specific V . parahaemolyticus strain No . 3525 belongs to the serological type 03:K30, which might be common to the sea waters of the Eastern United States . The possibility that fish serve as a reservoir to Vibrio parahaemolyticus and its possible implication in the transmission of human disease are discussed.

J Infect Dis, 1976 Jul, 134(1), 15 - 24
Challenge of dogs with live enterotoxigenic Escherichia coli and effects of repeated challenges on fluid secretion in jejunal Thiry-Vella loops; Sack RB et al.; Dogs were evaluated as experimental models for the study of diarrheal disease produced by enterotoxigenic Escherichia coli . Although a suitable whole model for orogastric bacterial challenge could not be developed, chronic jejunal Thiry-Vella loops were used to study the secretory effects of multiple jejunal challenges with enterotoxin of either Vibrio cholerae or E . coli . The heat-stable and heat-labile E . coli enterotoxins could be differentiated clearly in this model . Sequential weekly challenges over a four-week period showed a significant decrease in loop secretory response to homologous enterotoxin, although levels of antitoxin in serum remained unchanged, a finding suggesting a local immune response . Dogs challenged with E . coli enterotoxin were markedly protected against subsequent challenge with V . cholerae enterotoxin; the converse was not true . Histologic studies of the loops showed only minimal atrophy, and results of absorption studies in the loops were normal . These studies suggest that mongrel dogs are resistant to colonization by enterotoxigenic E . coli and partially resistant to challenge with enterotoxin, perhaps on an immune basis due to prior antigenic exposure . Multiple challenges with enterotoxin effect a decreased secretory response; this finding also suggests a local immune mechanism.

Lancet, 1976 Jun 12, 1(7972), 1258 - 61
Secretory IgA against enterotoxins in breast-milk; Stoliar OA et al.; A pool of colostrum from Guatemalan mothers (Guatemalan colostrum)) obtained 2-4 days post partum inhibited the induced fluid accumulation in rabbit ileal loops when incubated with Vibrio cholerae or Escherichia coli enterotoxin . There was a linear relationship between the quantity of colostrum used and the protection achieved . Pools of Guatemalan breast-milk obtained 15-30 days post partum and North American breast-milk had the same effect when tested with E . coli and V . cholerae enterotoxins, respectively . The antitoxic activity of a given pool correlated with its IgA content but not with the concentration of IgG or IgM . Guatemalan colostrum globulins were precipitated by ammonium sulphate . The globulins were filtered through a 'Biogel A5' column and fractions obtained . When tested in rabbit ileal loops the antienterotoxin activity in these fractions closely paralleled their IgA but not their detectable IgG or IgM content . We hypothesise that IgA antibody to enterotoxin, present in breast-milk of normal mothers, is probably a manifestation of natural immunity . The passive transfer of these antibodies to the infant may explain why breast-milk prevents E . coli diarrhoea in the neonate.

J Natl Cancer Inst, 1976 Jun, 56(6), 1113 - 8
Humoral reponse to neuraminidase-treated tumor cells; Sansing WA et al.; L1210 leukemia cells grew progressively and caused tumor deaths in all recipient mice . However, when these cells had been treated with Vibrio cholerae neuraminidase (VCN) prior to injection, tumor deaths did not occur . Both untreated and VCN-treated L1210 cells elicited a humoral response, as manifested by an increasing percent of cells in the spleen and peritoneal cavity, with various types of membrane-associated immunoglobulins . Progressive tumor growth was associated with a large percent of peritoneal exudate (PE) cells bearing membrane-associated IgG, a late increase in the percent of PE cells with IgG, and only a small percent of PE cells with IgM on their surfaces . Conversely, PE cells from mice given VCN-treated L1210 cells were characterized by a small percent with IgG, an early increase in percent of cells with IgG, and a large percent with membrane-associated IgM . An injection of VCN-treated L1210 cells into mice with progressively growing L1210 tumors caused frequent tumor remissions, with a corresponding alteration of the ongoing humoral responses . Both the degree of alteration and the number of cures depended on the tumor burden at the time VCN-treated tumor cells were injected . The humoral response in mice with tumor remission following immunization was comparable to the response detected after an injection of VCN-treated cells only.

J Gen Microbiol, 1976 Jun, 94(2), 367 - 72
Effect of growth temperature and culture age on the lipid composition of Vibrio cholerae 569B (Inaba); Raziuddin SR; The relative amounts of the major phospholipids (phosphatidylethanolamine, phosphatidylglycerol and lyso-phosphatidylethanolamine) and fatty acids in Vibrio cholerae 569B (Inaba) varied with growth temperature and between exponential and stationary phases of growth.

Zh Mikrobiol Epidemiol Immunobiol, 1976 Jun, (6), 94 - 6
{Study of the fractional makeup of diagnostic nutrient media for isolating Vibrio cholerae}; Golshmid VK et al.; A study was made of the fractional composition of peptones used for preparation of solid nutrient media . It was shown that the best growth of the vibrio on these media was obtained with the use of highly split peptones . Examination of fractional composition of peptone by gel-filtration and fractionation with various precipitants is recommended for rational working out of nutrient media for cultivation of cholera vibrio.

Zh Mikrobiol Epidemiol Immunobiol, 1976 Jun, (6), 82 - 4
{Polyvalent diagnostic cholera phage}; Drozhevkina MS et al.; A polyvalent diagnostic bacteriophage for rapid identification (in 6-10 hours) of cholera vibrio of both biotypes has been constructed of virulent E1 Tor phages and recommended for laboratory practice . Polyvalent diagnostic bacteriophage is strictly specific, inactive against the nonagglutinating vibrios, microbes of enteric group and those closely affiliated.

Am J Vet Res, 1976 Jun, 37(6), 737 - 40
Failure of vaccines to protect salmon from vibriosis enzootic in Puget Sound, Washington; Gunnels RD et al.; Juvenile chinook salmon in fresh water were vaccinated either orally or parenterally with heat- or formalin-killed bacterins prepared with Vibrio anguillarum . Subsequent exposure to naturally occuring vibriosis in the marine waters of Puget Sound (Washington State) indicated that no protection resulted . It was demonstrated, however, that protection could be achieved with the passive transfer of immune serum.

Med Microbiol Immunol (Berl), 1976 Jun 1, 162(2), 153 - 8
Suppression of growth of Vibrio cholerae in the intestine of immunized mice; Knop J et al.; In ligated intestinal loops of actively immunized adult mice, growth of V . cholerae 569B was suppressed approximately seven fold when compared to bacterial growth in non-immune animals . Similarly, growth of V . cholerae 569B was reduced in mice immunized with a hybrid vibrio strain, NCV569B-165, which shares only flagella antigens with V . cholerae 569B . Immunofluorescence studies of intestinal loop contents and intestinal sections indicated that, in non-immune mice, vibrios coated the intestinal mucosa . In contrast, the intestinal mucosa of immune animals was almost free of bacteria . The vibrios were found agglutinated in the intestinal lumen contents of immune animals . It was concluded that immunization suppressed the growth of V . cholerae in the intestinal lumen and the possibility that bacterial agglutination mediates this growth suppression is discussed.

Med Microbiol Immunol (Berl), 1976 Jun 1, 162(2), 133 - 6
Butandioldehydrogenase in Vibrio parahaemolyticus; Schroter G et al.; 396 strains of V . parahaemolyticus, 345 originating from Togo and 51 from Japan, were submitted to the study of butandiol dehydrogenase (BDH) . It was found that 52.3% of the strains were BDH-positive and 44.2% BDH-negative . An intermediate group characterized by weak reactions amounted to 3.5% of the strains . The distribution of this enzyme parallels to a certain degree the results of serotyping . Positive reactions have been found in all or most strains belonging to serotypes 03K29, O4K8, 04K12, 04K55, 05K15, and 012K19, whereas strains of serotypes 03K5 and 04K10 gave negative reactions . In serotype 05K17 both properties were found in nearly equal proportions . The test of BDH, which can supplement serotyping in characterizing strains of V . parahaemolyticus, can be applied as an aid to epidemiological studies.

Zh Mikrobiol Epidemiol Immunobiol, 1976 Jun, (6), 89 - 94
{Comparative study of the proteinograms of vibrios and of closely related microorganisms by means of polyacrylamide gel disc electrophoresis}; Atarova GT; Proteinograms of 112 strains of vibrios and closely affiliated microorganisms were studied by disc electrophoresis in polyacrylamide gel . Up to 25 protein peaks with definite mobility coefficients were revealed . The influence of the culture medium on the protein spectrum of the microbes was found . The frequency of peak formation was of great significance for the differentiation of the microbes under study . The quantitative characteristics of the peak area could not be used for differentiation.

Recent Adv Stud Cardiac Struct Metab, 1976 May 26-29, 11, 621 - 5
Increase in membrane conductance and positive inotropic action of hemolysin produced by Vibrio parahaemolyticus on rabbit myocardium; Seyama L et al.; The effects of purified thermostable direct hemolysin from the cultured filtrate of Vibrio parahaemolyticus on both S-A node cells and atrial cells were studied electrophysiologically . Hemolysin caused the membrane conductance to increase, thereby causing membrane depolarization . The major ion responsible for this depolarization was Na+, but other ions, such as K+ and Ca2+, also participated . Positive inotropic action of hemolysin was observed, and the Na-Ca exchange mechanism was enhanced after hemolysin treatment.

Recent Adv Stud Cardiac Struct Metab, 1976 May 26-29, 11, 615 - 20
Stopping of spontaneous beating of cultured mouse and rat myocardial cells by a toxin (thermostable direct hemolysin) from Vibrio parahaemolyticus; Goshima K et al.; Low concentrations of thermostable direct hemolysin from Vibrio parahaemolyticus stopped the spontaneous beating of cultured mouse and rat myocardial cells . These low concentrations depolarized the maximal diastolic potential and inhibited the generation of action potential of cultured myocardial cells . The toxin lost its activity when preincubated with ganglioside, GT1 or GM1 . GT1 was more effective than GM1 . High concentrations of the toxin caused morphological damage of cultured mouse and rat myocardial cells, but did not stop the beating, or cause morphological damage of cultured chick myocardial cells.

Antibiotiki, 1976 May, 21(5), 460 - 3
{Clinical evaluation of antibiotic therapy of NAG-infection}; Khabiblov SB et al.; It is not concluded yet whether it is expedient to use antibiotic therapy with respect to patients and vibrio-carries with NAG-infection . Observation of a group of patients with acute gastro-intestinal infections caused by NAG-vibrio and carriers of NAG-vibrioes showed that the rate of vibrio isolation after a course of antibiotic therapy (tetracycline, levomycetin) significantly decreased as compared to that in the group of the patients subjected only to symptomatic therapy . The data of the study provided recommendation of antibacterial therapy with respect to patients with NAG-infection especially in cases with accompanying infections or invasions . As for "asymptomic" carriers antibiotic therapy is required only with respect to persons with repeated vibrio isolation.

Mikrobiologiia, 1976 May-Jun, 45, 547 - 51
{Electron microscopic study of the Rybinsk water reservoir microflora}; Lapteva NA; Microflora of the Rybinsk water reservoir was studied by electron microscopy throughout the year . Common and rare bacterial forms were revealed which had not been detected before by light microscopy . The following forms prevailed: rod-like, coccoid, and bacteria of the vibrion type . Rare species were confined to certain places of the reservoir and related to seasonal changes in it . Bacteria belonging to the Planktomyces genus were registered in the central part of the reservoir in July--August . Swamp waters always contained bacteria belonging to the Hyphomicrobium genus . Spirillar bacterial forms were constantly present in coastal waters.

Biull Eksp Biol Med, 1976 May, 81(5), 561 - 4
{Model of a chronic Vibrio cholerae carrier based on gnotobiotic rats}; Baroian OV et al.; Germ-free monoflora (contaminated with nonpathogenic spore-bearing bacillus) and common albino rats (OFA) were infected with V . cholera El Tor, of Ogava and Inaba serological types (6 milliard microbial cells per 1.5 ml of physiological solution per rat) . Approximately one week after the infection the number of vibrios reached hundreds of millions per 1 g of feces and persisted at this level for over 100 days (observation period) . Newborn rats were infected by natural way from adult vibrio carriers and also became vibrio-carriers . The number of vibrios excretedby the animals which were germ-free and monoflora before the infection was approximately the same . Cholera vibrios perished completely in the enteric tract and were never revealed in the feces of common (control) rats.

Biull Eksp Biol Med, 1976 May, 81(5), 556 - 8
{Choleragen and neuraminidase production by Vibrio in vitro}; Durikhin KV et al.; In vitro studies demonstrated a high degree of the rank correlation between the synthesis of cholera exotoxin and neuraminidase by cholera vibrios (V . cholerae 569B) . The appearance of these biochemically-active materials proved to depend on the growth phase of the microbial population . A possibility of cooperation between them in the pathogenesis of cholera is suggested.

Zh Mikrobiol Epidemiol Immunobiol, 1976 May, (5), 32 - 7
{Immunochemical and biochemical characteristics of a preventive preparation against cholera, cholerogen-toxoid}; Dzhaparidze MN et al.; Cholerogen-toxoid served as a complex vaccine preparation composed of the main pathogenicity factors of cholera vibrio--cholerogen (toxoid), endotoxin and a number of exoenzymes . The preparation contains 65 +/- 7.5% of protein, 12 +/- 1.2% of reducing sugars, 7 +/- 1.2% of lipids, and 2 +/- 0.3% of nucleic acids . Analytic disc-electrophoresis in polyacrylamide gel and immune disc-electrophoresis revealed at least seven individual proteins with the serological activity in the preparation . About 70% of these constituted toxoid proper; the content of O-antigen was 22% . In the cholerogen-toxoid there were revealed seven exoenzymes of cholera vibrio; proteinase, lecithinase, lipase, DNA-ase, RNA-ase, hyaluronidase, amylase . Antibodies against proteinase, lecithinase, amylase and RNA-ase of cholera vibrio were found in the serum of rabbits immunized with cholerogen-toxoid.

Infect Immun, 1976 May, 13(5), 1467 - 72
Temperature-dependent inactivating factor of Pseudomonas aeruginosa exotoxin A; Vasil ML et al.; The adenosine diphosphate ribosyl transferase activity of Pseudomonas aeruginosa exotoxin A(PA toxin) was found to be rapidly destroyed by heating at 45 to 60C but not by heating at 70 to 90C (for at least 30 min) . This phenomenon has been previously described for other bacterial toxins (staphylococcal alpha-toxin and Vibrio parahaemolyticus hemolysin) and is termed an Arrhenius effect . In contrast, the Arrhenius effect was not seen when the PA toxin was heat-treated as above and tested for cell toxicity or mouse lethality . Although the PA toxin treated at 70C for 30 min retained a significant proportion (is greater than 70%) of its adenosine diphosphate ribosyl transferase activity, the cell toxicity and mouse lethality of the toxin were virtually abolished . A temperature-dependent inactivating factor that has proteolytic activity and is co-purified with the PA toxin was shown to be responsible for the Arrhenius effect . PA toxin separated from the factor by conventional disc gel electrophoresis or PA toxin preparations lacking the factor did not show the Arrhenius effect.

Med Klin, 1976 Apr 9, 71(15), 629 - 30
{Appearance of anaerobic vibrionen species (vibriones) in cases of chronically recurrent vaginal fluor (author's transl)}; Weissenbacher ER et al.; In 480 patients suffering from chronically recurring and/or therapy-resistant fluor of the outer female urogenital tract, a vaginal virbiosis was stated in 54 cases as cause of the fluor complaints lasting, for the most part, longer than five years . Aspects of vaginal vibriosis and respective therapy are discussed.

J Infect Dis, 1976 Apr, 133(4), 436 - 40
False-positive rabbit ileal loop reactions attributed to Vibrio parahaemolyticus broth filtrates; Johnson DE et al.; Vibrio parahaemolyticus broth filtrates have previously been shown to produce positive reactions in rabbit ileal loops only if concentrated 10-fold by lyophilization . This method of concentration produces solutions that contain greater than 20% NaCl . In the present study, however, concentrations of NaCl of greater than or equal to 4% induced positive responses in ileal loops, and desalting rendered previously reactive, concentrated broth filtrates negative . Therefore, enterotoxin was not demonstrated in our broth filtrates of V . parahaemolyticus, a finding which suggests that previous studies require further evaluation . Since most culture media contain 0.5% NaCl, it is important to determine and to control the NaCl content and the osmolality of all lyophilized concentrates tested in the ligated rabbit ileum.

J Bone Joint Surg Am, 1976 Apr, 58(3), 312 - 7
Immunological studies in murine osteosarcoma . Immunogenicity, growth kinetics, and immunotherapy; Miller CW et al.; A transplantable murine osteosarcoma is described . Following transplantation into a syngeneic mouse the tumor grows rapidly and kills the mouse with pulmonary metastases simulating human osteosarcoma . A cell-mediated antibody response is evoked in the host mouse as demonstrated by in vivo and in vitro tests . The number of pulmonary metastases may be decreased with adjunctive immunotherapy following excision of the primary tumor . Immunotherapeutic materials include BCG and isologous cells treated with Vibrio cholerae neuraminidase.

Hoppe Seylers Z Physiol Chem, 1976 Apr, 357(4), 559 - 66
Demonstration of neuraminidase activity in human blood serum and human milk using a modified, radioactively labelled alpha1-glycoprotein as substrate; Schauer R et al.; N-Acetylneuramini acid of alpha1-glycoprotein was oxidized with a small molar excess of periodate and reduced with tritium-labelled borohydride . By this method about 50% of the N-acetylneuraminic acid was converted to its radioactively labelled C8-analog and 25% to its C7-analog . Using this modified alpha1-glycoprotein as substrate, minimum neuraminidase concentrations of 10(-18) units/ml, related to the activity of neuraminidase from Vibrio cholerae, could be determined . Neuraminidase activity was demonstrated in 95% of the sera or blood plasma samples from a series of 417 healthy or ill human individuals and in milk samples from 5 different donors . The neuraminidase in both serum and milk had optimal activity at pH 5.5 . On an average, 10(-10) neuraminidase units were found in 1 ml serum and 10(-8) units in 1 ml milk . Although the neuraminidase activities in the sera varied, a correlation with definite pathological states is not yet possible . N-Acetylneuraminate pyruvate-lyase activity could not be detected in human serum.

J Clin Microbiol, 1976 Apr, 3(4), 432 - 7
Clinical isolates of anaerobic gram-negative rods with a formate-fumarate energy metabolism: Bacteroides corrodens, Vibrio succionogenes, and unidentified strains; Smibert RM et al.; Strains of anaerobic, gram-negative bacteria, isolated from human clinical specimens and from studies of human normal flora, that have energy metabolism similar to Vibrio succinogenes are described . Included are four human isolates of V . succinogenes, five similar strains of motile straight rods, three strains of Bacteroides corrodens, and two unidentified strains . All strains studied grew poorly in usual anaerobic broth media but produced good turbidity in overnight broth cultures in media containing fromate and fumarate, indicating that all have an energy metabolism similar to V . succinogenes: they gain energy by the transfer of electrons from formate or hydrogen to fumarate.

Isr J Med Sci, 1976 Apr-May, 12(4-5), 288 - 303
Characteristics of immunity induced by neuraminidase-treated lymphosarcoma cells in C3H (MTV+) and C3H (MTV-) mice; Bekesi JG et al.; The immunogenicity of 6C3HED lymphosarcoma ascites tumor cells was greatly enhanced upon treatment with Vibrio cholerae neuraminidase . As a result, mammary tumor virus (MTV)-free C3H mice which had been repeatedly immunized with the treated cells resisted a challenge of 10(6) untreated viable cells inoculated i.p., whereas all control animals died after receiving less than 10 lymphosarcoma tumor cells . In contrast, there was no increase in the refractoriness to challenge when MTV-infected C3H mice were immunized . Incubation of 6C3HED lymphosarcoma cells with serum of splenic lymphocytes from the immunized MTV-free C3H mice neutralized their tumorigenicity and protected recipient MTV-free and MTV-infected C3H mice against the disease . As shown by the 51Cr microcytotoxicity assay, the complement-dependent cytotoxic antibody titer reached 1,024; 29% of the target cells were lysed by lymphocytes obtained from immune MTV-free C3H mice . When the immune lymphocytes were pretreated with anti-theta serum they lost their ability to neutralize the tumorigenicity of the lymphosarcoma, failed to be stimulated by the T cell mitogens, phytohemagglutinin and concanavalin A, and failed to achieve cytolysis of the lymphosarcoma target cells in vitro . These observations establish the central role of the theta-antigen-bearing immune lymphocytes in cell-mediated immunity in this system . Failure of the immune system in the immunized MTV-infected C3H mice to cope with the challenging lymphosarcoma, even at low cell numbers, was reflected, not in the absence of humoral response, but in the relatively low level of cell-mediated immunity . This was manifested in both the neutralization and in the vitro cell-mediated immunity assays . This apparent immunological deficiency observed in the MTV-infected C3H mice could be countered by the transfer of lymphocytes from MTV-free C3H mice immunized with neuraminidase-treated 6C3HED lymphosarcoma cells . The recipient MTV-infected C3H mice then rejected a subsequent challenge with lymphosarcoma tumor grafts.

N Z Med J, 1976 Mar 10, 83(559), 155 - 6
Vibrio parahaemolyticus-food poisoning: case report; Corner BM et al.; Symptoms of food poisoning occurred following the ingestion of raw shellfish purchased in the Auckland area . Vibrio parahaemolyticus was recoverer from the patient . The potential of this and closely related microorganisms to cause illness is reviewed.

Zentralbl Bakteriol {Orig A}, 1976 Mar, 234(2), 212 - 8
Morphological, cultural and biochemical characteristics of Vibrio parahaemolyticus, isolated in Romania from acute gastro-enteritis; Ciufecu C et al.; Out of 399 human faeces examined during the first eight months (1975) for the presence of NCV vibrios, one vibrio parahaemolyticus strains has been isolated from a man with acute gastro-enteritis (gastric and abdominal pains, nausea, diarrhoea, headache, general weakness), after having a meal with salted caviar . The strain belongs to Heiberg's group VII and Sakazaki's subgroup I . The virulence tested on chick embryo is 3 times higher (LD50 = 14 germs) if compared with the virulence of V . parahaemolyticus control strain (Sakazaki's strains) . First isolation in Romania from a human stool (patient with acute gastro-enteritis).

Infect Immun, 1976 Mar, 13(3), 876 - 83
Cytotoxic effect of the thermostable direct hemolysin produced by Vibrio parahaemolyticus on FL cells; Sakurai J et al.; The thermostable direct hemolysin produced by Vibrio parahaemolyticus showed cytotoxic activity on FL cells derived from human amniotic membrane . Scanning electron micrographs of the whole cells showed that the microvilli on the cell surface decreased in number and changed in shape on treatment with the hemolysin . Most of the microvilli disappeared before death of the cells, as judged from the results of staining the cells with trypan blue and measuring release of alkaline phosphatase from the cells . Electron micrographs of thin sections ofthe cells showed that the cytoplasm of the cells was not significantly affected by treatment with sublethal amounts of hemolysin, even when the microvilli on the cell surface were significantly affected . Lethal amounts of hemolysin affected the cytoplasm and caused disarovilli on the cell surface are affected by treatment with the hemolysin before cytoxic effects develop.

Infect Immun, 1976 Mar, 13(3), 735 - 40
Synergistic protective effect in rabbits of immunization with Vibrio cholerae lipopolysaccharide and toxin/toxoid; Svennerholm AM et al.; Subcutaneous immunization of rabbits with a combination of Vibrio cholerae lipopolysaccharide (LPS) and enterotoxin induced a more than 100-fold-higher degree of protection against intestinal challenge with live cholera vibrios than did vaccination with either of the two antigens alone . Such a synergistic effect was also obtained by immunization with a combination of LPS and choleragenoid . The immunization with LPS and toxin (or toxoid) in combination did not enhance the reistance to toxin challenge above that induced by the toxin component alone . This, together with data from titrations of anti-LPS and antitoxin antibodies in serum and in intestinal washings, contradicts enhanced immune responses due to adjuvant action of the two antigens as the explanation for the synergistic effect of the combined vaccines . A more likely explanation would be that the antibacterial and antitoxic immune responses, without being increased in themselves, function synergistically by interfering with two separate events in cholera pathogensis.

J Infect Dis, 1976 Mar, 133 Suppl, 82 - 8
Stimulation of adenylate cyclase by Vibrio cholerae toxin and its active subunit; Berkenbile F et al.; The mechanism by which Vibrio cholerae toxin and its active subunit A stimulate adenylate cyclase of rat liver plasma membrane after in vivo or in vitro exposure was investigated . Stimulation of adenylate cyclase was more efficient with cholera toxin than with subunit A in whole animals and in "soluble" preparations of adenylate cyclase . Toxin-stimulated plasma membrane adenylate cyclase activity persisted after solubilization from the membrane by detergent, while subunit A stimulation was destroyed by detergent treatment . Detergent-free "soluble" enzyme could be stimulated in vitro by cholera toxin, but such stimulation by subunit A was very weak . Binding studies with iodotoxin or iodosubunit A confirmed that the interaction of toxin with adenylate cyclase in "soluble" preparations could be the result of specific binding to the enzyme . The uncertainty as to whether G M1 ganglioside (galactosyl-N-acetylgalactosaminyl {sialosyl} lactosyl ceramide; GGnSLC) is present in these fractions limited the interpretation of the data on binding . A biological modification of subunit A prior to the activation of adenylate cyclase is inferred from these data.

J Infect Dis, 1976 Mar, 133 Suppl, 75 - 81
Aspects of the interaction of Vibrio cholerae toxin with the pigeon red cell membrane; King CA et al.; The ganglioside galactosy-N-acetylgalactosaminyl {sialosyl} lactosyl ceramide (GM1) is readily accumulated in pigeon red cell membranes soaked with {3H}GM1 (1-100 mug/ml) at 37 C for 30 min . This treatment enhances the activation of adenyl cyclase by the toxin of Vibrio cholerae . An attempt was made at correlation of the amount of incorporated GM1 with the increased binding of toxin and activation of adenyl cyclase . Cells with less than 2 mug of incorporated GM1 per 4 X 10(9) cells bind 5-10 mug more toxin than do untreated cells, which bind 0.25 mug per 4 X 10(9) cells . Cells with more than 2 mug of GM1 bound (per 4 X 10(9) CELls) which has been incorporated from micellar solutions of GM1 (greater than 20 mug/ml), do not bind any more extra toxin . In untreated cells, 0.1 mug of toxin is involved in the activation of adenyl cyclase . In GM1-treated cells 0.25-0.5 mug of toxin is involved, although at least 5 mug of toxin is bound . It is concluded that 90% of the extra toxin-binding sites on the GM1-treated cell are nonproductive.

J Infect Dis, 1976 Mar, 133 Suppl, 14 - 22
Chemical characterization of the structure of cholera toxin and its natural toxoid; Kurosky A et al.; Acrylamide gel electrophoresis demonstrated that the toxin of Vibrio cholerae is comprised of three polypeptide chains, alpha, beta, and gamma, of molecular weights 24,000, 9,700, and 9,700 daltons, respectively . Amino acid sequence analysis of intact toxin indicated a molecular composition of alpha gamma beta4 . Acrylamide gel electrophoresis and sequence analysis confirmed that the natural toxoid (choleragenoid) is identical to the toxin beta-chain . The alpha- and gamma-chains of the toxin are disulfide-linked (fragment A) but are noncovalently bound to the beta-chains . About 50% of the primary structure of the N-terminal portion of the beta-chain has been identified and a small segment of the D-terminus has also been characterized . Twenty residues of the N-terminal portions of the alpha- and gamma-chains have been tentativly identified . The amino acid composition of the beta-chain was determined and compared to that of the natural toxoid.

Appl Environ Microbiol, 1976 Mar, 31(3), 389 - 94
Relationships between heat resistance and phospholipid fatty acid composition of Vibrio parahaemolyticus; Beuchat LR et al.; Vibrio parahaemolyticus was grown in tryptic soy broth (TSB) containing NaCl levels of 0.5, 3.0, and 7.5% (wt/vol) . Cultures incubated at 21, 29, and 37 C were harvested in late exponential phases and thermal death times at 47 C (D47 c; time at 47 C required to reduce the viable population by 90%) were determined in phosphate buffer containing 0.5, 3.0, and 7.5% NaCl . At a given NaCl concentration in the growth medium, D47 c values increased with elevated incubation temperatures and with elevated levels of NaCl in the heating menstrua . Differences in thermal resistance of cells cultured at a particular temperature were greater between those grown in TSB containing 0.5 and 3.0% NaCl than between those grown in TSB containing 3.0 and 7.5% NaCl . D47c values ranged from 0.8 min (grown at 21 C in TSB with 0.5% NaCl) to 6.5 min (grown at 37 C in TSB with 7.5%, heated in 7.5% NaCl buffer) . Methyl esters of major phospholipid fatty acids extracted from cells were quantitated . The ratio of saturated to unsaturated fatty acids in cells grown at a given NaCl concentration increased with elevated incubation temperature . At a particular growth temperature, however, saturated to unsaturated fatty acids ratios were lowest for cells grown in TSB containing 3.0% NaCl.

J Infect Dis, 1976 Mar, 133 Suppl, 5 - 13
The subunits of cholera toxin: structure, stoichiometry, and function; van Heyningen S; The toxin of Vibrio cholerae dissociates into subunit A and an aggregate of subunit B (choleragenoid); the dissociation is rapid under denaturing conditions and slow at neutral pH . Subunit A has a molecular weight of 27,000 daltons (measured by sedimentation equilibrium or gel chromatography) and has two polypeptide chains (mol wt, approximately 22,000 and 5,000 daltons) joined by disulfide bonds . The molecular weight of subunit B in 6 M guanidine hydrochloride is 14,000 daltons when determined by sedimentation equilibrium or gel chromatography, although dodecylsulfate gel electrophoresis suggests a lower value . These results suggest a structure of AB4 for the toxin; studies of cross-linking with methyl-4-mercaptobutyrimidate confirm this structure . The properties of antibodies both to cholera toxin and to choleragenoid are compatible with this structure, but subunit A has very low immunogenicity . Subunit A by itself is active, and this activity is abolished by a large excess of antitoxin but not by choleragenoid, anticholeragenoid, or ganglioside GM1 (galactosyl-N-acetylgalactosaminyl {sialosyl} lactosyl ceramide; GGnSLC) . It is suggested that the function of subunit B is not to interact directly with the adenylate cyclase system, but to bind to the cell membrane and facilitate the interaction of subunit A.

J Infect Dis, 1976 Mar, 133 Suppl, 23 - 30
Chemistry of cholera toxin: the subunit structure; Lai CY et al.; The toxin of Vibrio cholerae was separated into two subunits by gel filtration on Sephadex G-75 in 5% formic acid . The subunits were designated A and B . Amino acid analysis indicated that subunit B corresponded to choleragenoid . Renatured subunit B was found to be antigenically identical to the whole molecule, whereas renatured subunit A was not . On reduction and S-carboxymethylation with {2-14C} iodoacetate in 8 m urea, subunit A separated into two polypeptides of unequal sizes, A1 and A2, with equal amounts of radioactivity . Subunit B, on the other hand, remained a single molecular species . Molecular weights of the polypeptides A1, A2, and B were estimated by polyacrylamide gel electrophoresis in sodium dodecyl sulfate-urea buffer, in conjunction with amino acid analysis, to be 20,000, 7,500, and 9,500 daltons, respectively . The carboxyl terminal sequence of subunit B was found to be -Met-Ala-Asn . After treatment of toxin with cyanogen bromide, the peptide Ala-Asn could be isolated and quantitated . From this data the number of B polypeptides in a molecule of toxin was estimated as six.

J Infect Dis, 1976 Mar, 133 Suppl, 142 - 56
Escherichia coli enterotoxin: purification, partial characterization, and immunological observations; Dorner F et al.; Enterotoxin, a diarrhea-inducing protein elaborated by pathogenic Escherichia coli strains, was isolated from the supernate of fermenter cultures of E . coli strain P263, a porcine enteropathogen . Purification involved chromatography and preparative isotachophoresis . The resulting product appeared to be pure according to immunoelectrophoretic, disc electrophoretic, ultracentrifugal, and immunologic criteria . The enterotoxin had an apparent molecular weight of 102,000 daltons, and its isoelectric point was 6.90 . The isolated product was active in inducing experimental diarrhea in adult rabbits and piglets . In small dosage it also elicited a drastic increase in adenylate cyclase activity in broken-cell preparations of cat heart tissue . The enterotoxin activity was acid labile and was destroyed by heat (65 C for 30 min) . It is suggested that the heat-stable enterotoxin was derived from heat-labile enterotoxin by complexing with endotoxin or with capsular material in the culture supernatant . The antigenic relations between the heat-labile enterotoxins of enteropathogenic E . coli strains of different serological types and different host adaptations, as well as between the E . coli enterotoxin and that of Vibrio cholerae, were investigated.

J Infect Dis, 1976 Mar, 133 Suppl, 138 - 41
Purification of heat-labile enterotoxin from Escherichia coli O78:H11 by affinity chromatography with antserum to Vibrio cholerae toxin; Dafni Z et al.; Concentrated culture filtrate of Escherichia coli O78:H11, strain H10407, was applied to an affinity column prepared with IgG antibodies to the toxin of Vibrio cholerae . Elution of the retained material with 3 M KCNS yielded a nonenterotoxic protein that precipitated with antiserum to V . cholerae toxin and had three major protein components on sodium dodecyl sulfate gels . After treatment with 2-mercaptoethanol, two protein components were observed . Elution of the affinity column with 5 M guanidine yielded an enterotoxic protein that precipitated with antiserum to V . cholerae toxin . After treatment with 2-mercaptoethanol, only one protein component, with mobility identical to that of the slower component of the eluate (treated with 3 M KCNS and 2-mercaptoethanol), was observed.

J Infect Dis, 1976 Mar, 133 Suppl, 120 - 37
Isolation and properties of heat-labile enterotoxin(s) from enterotoxigenic Escherichia coli; Finkelstein RA et al.; Various techniques have been applied to the detection of skin reactivity associated with heat-labile Escherichia coli enterotoxin in fermenter-grown cultures of enterotoxigenic strains in syncase medium and in trypticase soy broth . Isolated products that were homogeneous, as determined by disc electrophoresis and sodium dodecyl sulfate gel electrophoresis, differed immunologically and in physicochemical characteristics depending on the strain and medium used, even though the products had similar specific activities in skin tests and in Chinese hamster ovary cells . In support of observations on porcine E.coli enterotoxin, and in contrast to the enterotoxin of Vibrio cholerae, the products were single polypeptide chains with molecular weights ranging from approximately 35,000 ot over 100,000 daltons . Proteolytic cleavage during culture and purification might account for some of the variations observed . The isolated products were almost 10(6)-fold less active than purified choleragen in causing morphologic alterations of Chinese hamster ovary cells, approximately 1,000-fold less active in skin tests, and at least 100-fold less active in rabbit ileal loops . In addition, only 1/100 as much active protein was produced by the strains employed as by V . cholerae . It is possible that accessory or host-derived factors are required . The most effective large-scale procedure for isolation from trypticase soy broth culture supernatants was a sequence of concentration by ultrafiltration, judicious (NH4)2SO4 precipitation, gel filtration on Sephadex G-150, and gel filtration on Agarose A5m, from which the E . coli products (like choleragen) are retarded in their elution . Toxin was isolated from an enterotoxigenic strain that caused diarrhea (total volume, 60-liters) for four days in a patient who had visited Mexico.

J Infect Dis, 1976 Mar, 133 Suppl, 115 - 9
Interactions of choleragenoid and GM1 ganglioside with enterotoxins of Vibrio cholerae and Escherichia coli in cultured adrenal cells; Donta ST; The heat-labile enterotoxins of Vibrio cholerae and Escherichia coli induce morphologic changes and steroidogenesis in clonal lines of adrenal tumor cells in tissue culture; these effects are preventable by prior incubation of either toxin with GM1 ganglioside (galactosyl-N-acetylgalactosaminyl {sialosyl} lactosyl ceramide; GGnSLC) but are not preventable by prior incubation of adrenal cells with choleragenoid . Choleragenoid, however, is capable of interfering with the ability of GM1 ganglioside to neutralize the effects of either toxin . These results suggest that the GM1 ganglioside may not be the true receptor for the toxins on adrenal cells, but that it is acting as a pseudoreceptor . The ability of a subunit (A) of the cholera enterotoxin molecule to induce in adrenal cells morphologic changes and steroidogenesis similar to those effects inducible by the whole toxin indicates the possibility that separate receptor sites on these adrenal cells may exist for the binding and active fragments of the molecule.

J Infect Dis, 1976 Mar, 133 Suppl, 89 - 96
Interaction of Vibrio cholerae toxin with sarcoma 180 cell membranes; Wheeler MA et al.; Three discrete phases are discernible in the activation, by Vibrio cholerae toxin, of adenylate cyclase in fragments of sarcoma 180 cell membranes . In the first, or preparatory, phase the toxin must be exposed to dithiothreitol or nicotinamide adenine dinucleotide (NAD) in the absence of the membranes . In the second phase, the prepared toxin is dissociated to yield a macromolecular cyclase-activating factor (MCAF) in the presence of the membranes . In the third phase, membrane basal adenylate cyclase is activated by MCAF in the presence of NAD . The integrity of the catecholamine or beta-receptor associated with sarcoma adenylate cyclase is irrelevant in the activation of cyclase by MCAF . This activation proceeds undiminished even if the beta-receptor is desensitized or blocked by propranolol.

J Infect Dis, 1976 Mar, 133 Suppl, 55 - 63
Multiple roles of erythrocyte supernatant in the activation of adenylate cyclase by Vibrio cholerae toxin in vitro; Gill DM; Peptide A1 of Vibrio cholerae toxin, nicotinamide adenine dinucleotide, adenosine triphosphate, and a soluble protein present in erythrocyte supernatant are required for the activation of pigeon erythrocyte ghost adenylate cyclase but are not required to maintain the activated state . The compounds are all required simultaneously, and when all are added to ghosts, adenylate cyclase activity increases at a linear rate without delay . Under optimal conditions significant activation of cyclase is given by less than one molecule of toxin per ghost . Intact cholera toxin may be inactive in vitro . There is a delay of about 1 min between the addition of intact toxin and the attainment of the final rate of increase of adenylate cyclase activity . During this period, glutathione reduces the disulfide bond between peptides A1 and A2 . The delay is eliminated if the toxin is reduced before addition . More A1 is liberated if the toxin is also denatured with sodium dodecyl sulfate.

J Infect Dis, 1976 Mar, 133 Suppl, 108 - 14
Mode of action of Vibrio cholerae enterotoxin in cultured adrenal tumor cells; Wishnow RM et al.; The effects of Vibrio cholerae enterotoxin on steroidogenesis and on formation of adenosine 3':5'-cyclic phosphate (cyclic AMP) in two adrenal tumor cell lines were compared . Steroidogenesis was half-maximal at concentrations of 1 ng of cholera toxin/ml in the mutant OS-3 cells and 3 ng of cholera toxin/ml in the parent Y-1 cells . At the end of an 8-hr incubation, toxin-induced formation of cyclic AMP in the mutant cell line was reduced by 90% . A molar ratio of GM1 ganglioside (galactosyl-N-acetylgalactosaminyl {sialosyl} lactosyl ceramide; GGnSLC) to cholera toxin of 3:1 caused half-maximal inhibition of steroidogenesis in both cell lines . When equine antiserum to choleragenoid was added to adrenal cells 15 min after cholera toxin, there was marked inhibition of cyclic AMP formation and of steroidogenesis . Pretreatment of Y-1 cells with adrenocorticotropin rendered them unresponsive to hormonal induction of cyclic AMP formation, but these cells had an unimpaired response to cholera toxin . These studies, utilizing two adrenal cell lines, suggest important differences between the mode of action of cholera toxin and that of adrenocorticotropin in cultured adrenal tumor cells.

J Infect Dis, 1976 Mar, 133 Suppl, 103 - 7
Mechanism of activation adenylate cyclase in vitro by polymyxin-released, heat-labile enterotoxin of Escherichia coli; Gill DM et al.; Heat-labile enterotoxic material released from Escherichia coli by polymyxin B activates the adenylate cyclase of pigeon erythrocyte ghosts in a time- and concentration-dependent manner . The activation requires nicotinamide adenine dinucleotide, adenosine triphosphate, and another component of the erythrocyte supernatant . The active species has a molecular weight of about 23,000-24,000 daltons, is inhibited by antibodies to the toxin of Vibrio cholerae, and is not irreversibly denatured by sodium dodecyl sulfate . Thus in many respects the active species from E . coli behaves the same as peptide A1 of cholera toxin.

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