Schweiz Med Wochenschr, 1976 Feb 28, 106(9), 308 - 9 {Exocrine pancreatic function, intestinal resorption and reaction to Vibrio cholerae infection in protein deficiency}; Gyr K et al.; Six Patas monkeys were fed a protein-free diet while 16 animals of the same species receoved a standard monkey diet . The protein-depleted monkeys showed reduced absorption of folic acid but not Dxylose, and 5 out of 6 exhibited depressed exocrine pancreatic function . When challenged with Vibrio cholerae, animals with reduced folic acid absorption and pancreatic enzyme production excreted the micro-organisms for a longer time. Infect Immun, 1976 Feb, 13(2), 457 - 63 Virulence factors involved in the intraperitoneal infection of adult mice with Vibrio cholerae; Eubanks ER et al.; Nonmotile mutants of Vibrio cholerae isolated from Ogawa, Inaba, and El Tor strains were less virulent than parent wild types when administered to adult mice intraperitoneally . The cells were suspended in 5% hog gastric mucin . Antitoxic immunity did not protect mice against this type of challenge, but a ribosomally derived vaccine did . Intraperitoneal injection of 10 50% lethal doses of enterotoxin (based on intravenous doses) was without toxic manifestations as were 10(10) heat-killed vibrios similarly administered, regardless of strain . Virulent organisms killed with formalin or ultraviolet irradiation were significantly lethal at a dose of 10(10) cells . Mice made tolerant to endotoxin were protected from death caused by an injection of 3 X 10(10) boiled cells, but they did not survive an injection of formalin-killed cells . It is believed that the cause of death in this animal model of cholera is dependent, at least in part, on a toxic heat-labile moiety closely associated with the vibrios. Arch Microbiol, 1976 Feb, 107(1), 71 - 3 Tubular structures of Vibrio psychroerythrus; D'Aoust JY et al.; The marine bacterium Vibrio psychoerythrus, an obligate psychrophile, contains uniformly dense rod-shaped organelles 10-15 nm wide and up to 1.3 mum long . These structures were frequently seen crossing the septum of dividing cells. Antibiotiki, 1976 Feb, 21(2), 141 - 4 {Use of media with antibiotics for isolating resistant forms of the cholera vibrios}; Libinzon AE et al.; Higher stability of resistance to tetracycline in the polyresistant strain of V . eltor under conditions of macroorganism as compared to nutrient media was found experimentally . To increase isolation of the resistant forms of the cholera vibrio it was recommended to use agar with tetracycline or other antibiotics depending on the particular case in addition to the routine media. Zh Mikrobiol Epidemiol Immunobiol, 1976 Feb, (2), 81 - 6 {So-called mucoid forms of vibrio cholerae}; Shafershtein DL et al.; The authors present materials indicating that examination of nonagglutinating vibrios in the agglutination reaction with cholera sera after boiling, according to Sakazaki recommendation (1971), did not provide recognition of mucoid forms of cholera vibrio and its differentiation from cholera-like vibrios of other serological groups . It is suggested that one should take into consideration morphological peculiarities of the culture, stability of agglutination with cholera O-serum and the results of reaction of antibody adsorption in identification of cholera vibrios in the mucoid form. C R Acad Sci Hebd Seances Acad Sci D, 1976 Jan 26, 282(4), 397 - 400 {Bactericidal activity of sea water and its microvibrio content}; Guelin A; The bactericidal effect of sea water on E . coli varies according to the location and depth of specimen . There are seasonal variations . With the elimination of the bacilli, tiny vibrios appear . Two strains have been isolated and kept alive with bacteria suspensions in sea water . A method to quantify them in culture or in sea water has been developed. MMW Munch Med Wochenschr, 1976 Jan 9, 118(2), 27 - 30 {The cholera syndrome - pathogenesis and pathogens (author's transl)}; Muller HE; The same pathomechanism underlies both classical cholera and the enteritides due to nonagglutinating (NAG) vibrios and a few types of E . coli . A heat-labile enterotoxin activates the adenocyclase . This stimulates the cells of the small intestine to secrete ions and water, the immune reaction in the intestine is retarded and the production of gastric juice is reduced . Neuraminidase potentiates these reactions by opening additional enterotoxin receptors . Endotoxin is a further pathogenetic factor . The individual pathogens can certainly be clearly distinguished microbiologically, but the possession of the same pathogenetic factors makes the transition seem rather fluid . For this reason it is certainly wrong to regard NAG vibrios as only harmless water vibrios. Biull Eksp Biol Med, 1976, 81(4), 452 - 4 {Reduction of the biological activity of Vibrio cholerae culture filtrates exposed to neuraminidase inhibitors}; Durikhin KV; Experiments were conducted on a model of edema of albino mouse paws; a study was made of the effect of neuraminidase inhibitors on the cholerogenic effect of a filtrate of cholera vibrio culture . It appeared that addition to the filtrate of inhibitors depressed its biological activity . Since no cholerogenic action was possessed by the purified neuraminidase preparations from the cholera vibrios it was suggested that there existed a chemical affinity between the region of cholerogen responsible for fixation on the cell membranes and the active neuraminidase centre. J Natl Cancer Inst, 1976 Jan, 56(1), 83 - 7 Antitumor immunity . II . Viability, tumorigenicity, and immunogenicity of neuraminidase-treated tumor cells: effective immunization of animals with a tumor vaccine; Ray PK et al.; A dimethylbenzdithionaphthene (DBDN)-induced fibrosarcoma showed reduced transplantability if previously treated with Vibrio cholerae neuraminidase (VCN) . However, the reduced transplantability of VCN-treated tumor cells was not associated with any loss of their viability or tumorigenic capability, but appeared to be due to their increase in immunogenicity . High doses of VCN-treated tumor cells could grow even in normal individuals . Lower doses, which did not induce tumor development in normal individuals, did so if injected into immunosuppressed animals . Although X-irradiation of VCN-treated tumor cells reduced their tumorigenic potential, it did not impair their increased immunogenic properties . Thus a suitable method for the preparation of a "tumor vaccine" was provided . The immunization of animals with the vaccine and a working hypothesis regarding its mechanism of action were described. Infect Immun, 1976 Jan, 13(1), 195 - 203 Distribution of cholera organisms in experimental Vibrio cholerae infections: proposed mechanisms of pathogenesis and antibacterial immunity; Schrank GD et al.; This study was undertaken to determine the sequence of events in the microenvironment of the intestinal tract that culminate in the symptoms of cholera and to attempt to define more clearly the mechanisms involved in antibacterial immunity . The extent to which mucus occurs in the normal intestine of rabbits and the appearance of the intestinal villi in unfixed frozen sections was demonstrated . The villi and intervillous spaces were found to be normally covered by a layer of mucoid material that formed a mucous zone between the intestinal contents and the tips of the villi . The distribution of cholera organisms in normal and immunized animals was demonstrated by the staining of frozen-tissue sections with specific fluorescent antibody . Study of tissue sections from normal animals showed that the onset of fluid accumulation was concomitant with the establishment of large masses of organisms in the intervillous spaces and crypts of the intestine after the successful penetration of this mucous zone . Tissue sections from animals actively or passively immunized against a cell wall antigen of Vibrio cholerae showed clumping of vibrios in the lumen and restricted distribution in the lumen and luminal border of the mucous zone . Antibody was not lytic in vivo. Infect Immun, 1976 Jan, 13(1), 163 - 71 Demonstration of the cardiotoxicity of the thermostable direct hemolysin (lethal toxin) produced by Vibrio parahaemolyticus; Honda T et al.; Intravenous injection of the thermostable direct hemolysin (lethal toxin) produced by Vibrio parahaemolyticus caused rapid death of rats . Studies by electroencephalography and electrocardiography showed that after intravenous injection of the toxin the electroencephalogram remained normal for quite a long time after the heart of the animals had stopped beating . Depression of intraatrial and intraventricular conduction of electrical activation, including atrioventricular block, was observed in electrocardiograms of animals injected with the toxin . The toxin was also found to be toxic to cultured mouse heart cells . When it was added to the medium, the beating rhythm of the cultured heart cells increased temporarily and then soon stopped abruptly . The effect of the toxin on cultured mouse heart cells was blocked by preincubation of the toxin with a ganglioside mixture . From these results it is concluded that the thermostable direct hemolysin (lethal toxin) had cardiotoxic activity, and thus administration of the toxin causes rapid death of animals. Infect Immun, 1976 Jan, 13(1), 133 - 9 Identification of lethal toxin with the thermostable direct hemolysin produced by Vibrio parahaemolyticus, and some physicochemical properties of the purified toxin; Honda T et al.; Lethal toxin was purified extensively from the culture filtrate of a Kanagawa phenomenon-positive strain of Vibrio parahaemolyticus . The purified toxin was a protein, and its homogeneity was demonstrated by sodium dodecyl sulfate-polyacrylamide gel disc electrophoresis and analytical ultracentrifugation . It was demonstrated that the thermostable direct hemolysin was identical to the lethal toxin and that it was the main, if not only, lethal toxin in the culture filtrate . The purified toxin had a lethal effect when injected into mice either intravenously or intraperitoneally . Its lethal effect was very rapid, a dose of 5 mug of toxin per mouse killing the animals within 1 min . The lethal activity was inhibited by a ganglioside mixture . Some physicochemical properties of the purified toxin are reported. Ciba Found Symp, 1976, (42), 129 - 47 Intestinal immunization with soluble bacterial antigens: the example of cholera toxoid; Pierce NF; The studies described are aimed at a better understanding of the intestinal immunological system and its role in protection against enteric infection . The cellular kinetics of the intestinal immune response to cholera toxoid were studied in rats and the protection afforded by toxoid immunization was studied in dogs . Memory was demonstrated in the gut immune system . Plasma cells containing IgA antitoxin appeared in large numbers in gut lamina propria when intraduodenal boosting followed either intraperitoneal priming or prolonged oral priming, intraperitoneal priming being the most efficient . Immunization by the intraperitoneal route alone produced no response in small bowel lamina propria . Lamina propria plasma cells were derived from precursors in Peyer's patches or mesenteric lymph nodes which migrated through the thoracic duct and systemic circulation before homing to the gut . Dogs were immunized parenterally with cholera toxin or toxoid and challenged orally with Vibrio cholerae . Protection correlated closely with serum antitoxin titres and was usually brief . Passive intravenous immunization with IgG antitoxin was also protective . In contrast, subcutaneous priming followed by oral boosting yielded longer protection without elevated serum antitoxin titres . Antitoxin was detected in jejunal washings only briefly after local boosting . The mechanism by which protection is prolonged is unclear but its greater duration after parenteral priming and oral boosting emphasizes the importance of stimulating the gut immune mechanism in attempts to immunize against enteric bacterial infections . The parenteral-oral squence may be an effective means of immunizing the intestine with non-replicating protein antigens. Arch Invest Med (Mex), 1976, 7(2), 57 - 60 {Induction of phagocytosis by immunotherapy against leukemic leukocytoblasts}; Gomez-Estrada H et al.; Since 1975, the authors have been working in immunochemotherapy in acute myeloblastic leukemic adult patients . Autologous leukemic leucocytoblasts have been cultured with Vibrio cholerae neuraminidase (NVC) . Thereafter, neoplastic cells were radiated with cobalt bomb and frozen to--70degreesC and afterwards 5 X 10(6) of these leucoblasts were intradermally injected to each patients every eight days . All patients received between six to ten injections . So far, ten such patients have been followed during eight months and since two months after immunization no leukemic leucocytoblasts in bone marrow and blood have been detected . Hemoglobin and platelets have increased more rapidly than usual and no patient has developed side effects . Phagocytosis tests carried out in vitro during remission, have demonstrated that only in immunized patients neutrophils and monocytes were capable of phagocytosis of autologous cultured malignant leucocytoblasts . The nucleus of phagocytized malignant cells was small and basophylic . These so far undescribed phagocytic cells were named post-immunization cells. J Hyg Epidemiol Microbiol Immunol, 1976, 21(3), 374 - 6 Human infections caused by V . parahaemolyticus in Czechoslovakia; Laznickova K et al.; Vibrio parahaemolyticus was isolated in two subjects with acute gastroenteritis . The patients' history included stay in Pakistan or India . They probably contracted the infection on board an aeroplane where they ingested cold meal containing sea products . The possibility of this conditioned pathogen occurring also in an inland country is pointed out. J Hyg Epidemiol Microbiol Immunol, 1976, 21(3), 374 - 6 Human infections caused by V . parahaemolyticus in Czechoslovakia; Laznickova K et al.; Vibrio parahaemolyticus was isolated in two subjects with acute gastroenteritis . The patients history included stay in Pakistan or India . They probably contracted the infection on board an aeroplane where they ingested cold meal containing sea products . The possibility of this conditioned pathogen occurring also in an inland country is pointed out. Zh Mikrobiol Epidemiol Immunobiol, 1976, (12), 66 - 9 {Phage typing of cholera vibrios with different sets of phages}; Drozhevkina MS et al.; A comparative study was made of two sets of Mukerjee and Drozhevkina-Arutyunova's bacteriophages in typing 514 strains of the El Tor vibrios and 45 strains of clasic biotype . It was shown that the Mukerjee or Drozhevkina-Arutyunova's phages could be used for the typing of cholera vibrios . The phages of the latter set prove to detect more phage types (18 against 11); they determine both the phage type and the biotype at the same time . The typing of cholera vibrios of both biotypes is possible, and the percentage of nontyping strains left is comparatively low (5.2 against 23.5 after Mukerjee) . A table of the phage correspondence was made; it permits to obtain comparable data in using any set of the typing phages. Zh Mikrobiol Epidemiol Immunobiol, 1976, (12), 61 - 5 {Long-term survival of E1 Tor cholera vibrios in naturally contaminated sewage}; Zaidenov AM et al.; El Tor cholera vibrios of Ogava serological type were revealed in the sewage of the locomotive shed for 15 months . In experiment with an oil catcher in naturally infected sewage El Tor vibrios survived 36 days, in storage of this sewage at the laboratory--39 days, in the artificially infected sewage of a settlement and of a milk plant--2 and 11 days, respectively, in the oil and disel fuel--14 months . Consequently, El Tor vibrio can survive in the sewage with a high oil product content for a long time. Zh Mikrobiol Epidemiol Immunobiol, 1976, (12), 56 - 61 {Experimental cholera in guinea pigs . II . Pathogenesis of experimental infection}; Kotenok IaF et al.; It was found by bacteriological, anatomo-pathological and histological studies that intrapulmonary administration to guinea pigs of a suspension of the cholera causative agent containing colloidal substances (peptone, gelatine, agar-agar) caused primary reproduction of vibrios in the pulmonary tissue and the pleural exudate . From the lungs the microbes penetrated by hematogenic route into the liver and the bile system and with the flow of the infected bile entered the small intestine . Intestinal affection by the type of specific enteritis developed as a result of intensive vibrio reproduction in the submucous layer. Biull Eksp Biol Med, 1976, 82(11), 1357 - 9 {Inhibiting effect of cholera vibrio neuraminidase in Rauscher mouse leukemia}; Barinskii IF et al.; A study was made of the inhibitory effect of neuraminidase of cholera vibrio in mouse Rauscher leukemia . It was shown that the processing of cells of the spleen of mice suffering from leukemia with neuraminidase (in a dose of 50 units/ml and more) significantly inhibited the capacity of these cells to induce leukemia in their injection to BALB/c mice . In the mentioned concentration neuraminidase injected repeatedly parenterally produced no therapeutic effect in Rauscher leukemia. J Hyg Epidemiol Microbiol Immunol, 1976, 21(2), 150 - 6 Food poisoning caused by parahaemolytic and NAG vibrios after eating meat products; Zakhariev Z et al.; At total of 15 subjects taken ill with food poisoining caused by parahaemolytic and NAG vibrios after eating boiled pork sausage and boiled veal stomach were examined . The meat products were secondarily contamined in the sausage factory due to the use od non-disinfected lake water which was found to be a natural medium of the vibrios . When the use of this water was prohibited, the contamination of the factory and the meat products disappeared and no further cases of food poisoning due to meat products were notified . The disease has a toxic character, runs a bening course and end after 2--3 days by restoration to full health. J Hyg Epidemiol Microbiol Immunol, 1976, 20(2), 150 - 6 Food poisoning caused by parahaemolytic and NAG vibrios after eating meat products; Zakhariev Z et al.; At total of 15 subjects taken ill with food poisoining caused by parahaemolytic and NAG vibrios after eating boiled pork sausage and boiled veal stomach were examined . The meat products were secondarily contamined in the sausage factory due to the use od non-disinfected lake water which was found to be a natural medium of the vibrios . When the use of this water was prohibited, the contamination of the factory and the meat products disappeared and no further cases of food poisoning due to meat products were notified . The disease has a toxic character, runs a bening course and ends after 2--3 days by restoration to full health. Zh Mikrobiol Epidemiol Immunobiol, 1976, (3), 60 - 5 {Antigenic structure of non-agglutinating vibrios and bacteria of the genus Aeromonas}; Vvedenskaia OI et al.; The authors carried out a comparative study of the antigenic structure of nonagglutinating vibrios and the respresentatives of Aeromonas genus . A number of common antigens (some of which were of protein origin) were revealed in the so-called enterotoxins of nonaggultinating vibrios and aeromonas and also in the cytoplasm and ribosomes of these strains . "Enterotoxin" of Aeromona s strain contained the factor of vascular permeability . The serum to "enterotoxin" of the Aeromonas strain neutralized the enterotoxin of nonagglutinating vibrio. Zh Mikrobiol Epidemiol Immunobiol, 1976 Jan, (1), 30 - 4 {Patterns in the body's immune response to administration of El Tor vibrio cholera toxin to experimental animals}; Korovkina GV et al.; A study was made of some regularities attending the mechanisms of antibody production in the organism of experimental animals after the administration of El Tor cholera vibrio toxin . As revealed, the indices of the immune response depended on the chosen model, the method of its determination, the method of administration and the amount of the toxin administered . Dynamics of the primary and secondary responses of the immunocompetent cells of mouse spleen was determined . With the action of the El Tor toxin on the lymphoid tissue of mice there were expressed the general biological regularities of the response -- the optimal amount of the antigen-antibody complexes caused activation of the antibody-producing cells; high doses depressed the most reactive immunocompetent Y-cells . Apparently, there existed a genetic determinination of the immune response of the organism to a definite antigen -- the toxin of the El Tor cholera vibrio . It is supposed that it is necessary to choose groups which mostly require cholera vaccination. Dev Biol Stand, 1976, 33, 102 - 7 Possibilities of immunization against cholera and related enterotoxic enteropathies; Finkelstein RA; Scientifically controlled field studies have established that parenterally administered killed vibrio vaccines or somatic antigen preparations offer only limited degrees of protection in certain population groups and have made it obvious that new approaches to the immunoprophylaxis of cholera are needed . It has now also been established that the symptoms of cholera result from the action of the cholera enterotoxin (choleragen) on the epithelial cells of the small intestine . Immulogically related enterotoxins have been incriminated in other newly recognized diarrheal diseases (e.g., those caused by Escherichia coli and "non-agglutinable" (NAG)vibrios) . Additionally, volunteer studies have shown that induced cholera results in rather solid and lasting immunity against homologous re-challenge thus proving that immunity against cholera is feasible . Although numerous studies have demonstrated the efficacy of purely antitoxic immunity in experimental animal models, the protective effect of parenterally administered glutaraldehyde treated toxoid in man has shown to be limited, at best . The protection attained following an attack of cholera suggests that local immune mechanism may be of predominant importance . Immunity has been stimulated, experimentally in mice, by toxin antigen administered per os on a single occasion . Choleragenoid, which is non-toxic but binds to the same receptors as cholera toxin, has been shown to provide immediate resistance as well as later immunity to toxin challenge . More ideal, however, would be a colonizing mutant of V . cholerae which elaborates a non-toxic cross-reactive material (CRM) like choleragenoid and which could stimulate local antitoxic as well as anti-vibrio immune mechanisms . A tox-mutant of V . cholerae which is avirulent for man has been shown to elicit substantial immunity in man but the ideal CRM-mutant has yet to be found. Am J Trop Med Hyg, 1976 Jan, 25(1), 201 - 2 Emergence of Vibrio parahaemolyticus as an important cause of diarrhea in Malaysia; Jegathesan M et al.; Vibrio parahaemolyticus was isolated from 47 of 2,699 stools examined primarily for the exclusion of cholera . All strains grew well in alkaline peptone water containing 0.5% NaCl and in Monsur's medium . Serotyping showed them to be of various types . Adults were mainly affected . The importance of looking for V . parahaemolyticus in clinical specimens from cases of diarrhea is emphasized. Adv Exp Med Biol, 1976, 71, 191 - 204 Interaction of cholera toxin and ganglioside G(M1); Svennerholm L; The exotoxin produced by Vibrio cholerae is rapidly and firmly bound to the outer membrane of mammalian cells . With simple in vitro and in vivo methods and very pure gangliosides and allied glycolipids we have demonstrated that the monosialosylganglioside GM1 is the natural receptor for the cholera toxin . This ganglioside binds the toxin with a high affinity and inactivates it . The inactive derivative, choleragenoid toxoid has the same affinity to GM1 as the toxin . Ganglioside GM1 was isolated from the small intestinal mucosa of man, pig and ox in amounts of 0.1, 2.0 and 43 nmoles per g mucosa, respectively . These very large differences in the ability of the mucosal cells to bind cholera toxin . Exogenous GM1 was incorporated in vitro and in vivo in intestinal mucosal cells . The incorporation of GM1 increased the number of toxin-binding sites and increased the secretion of fluid in the gut . Vibrio cholerae sialidase did not hydrolyse the di- and trisialogangliosides of intact mucosal cells to the parent GM1-ganglioside, neither did it increase the number of cholera toxin-binding sites. J Med Primatol, 1976, 5(5), 305 - 11 Malnutrition and susceptibility to infection with Vibrio cholerae in vervet monkeys (Cercopithecus aethiops) . 2 . Response of vervet monkeys on protein, B-vitamin complex and calorie-deficient diets to infection; Felsenfeld O et al.; Five groups of vervet monkeys (Cercopithecus aethiops) were fed regular monkey chow, low protein (LP), low B-vitamin complex (LBV), LP-LBV, or low protein- low calorie (LP-LC) diets, respectively . After eight weeks the animals were infected with cholera vibrios . IgG, IgA and IgM increased in the sera of all animals after the challenge but the avidity of the immunoglobulins was lower in vervets on LP diets . Diarrhea and excretion of vibrios lasted longer in animals consuming less protein. Vet Med (Praha), 1976, 21(8), 483 - 9 {Biochemical characteristics of Vibrio coli isolated from inflammatory pseudomembraneous swine dysentery}; Vladik P et al.; The paper presents the basic morphological and biochemical characteristics of seven microaerophilic vibrio cultures isolated by the membrane filtration method from the colon mucous membrane of pigs diseased with dysentery . For growth, the cultivated strains needen oxygen reduction; in the dark field they showed a typical active movement producing catalase and oxidase, reducing nitrates to nitrites, and tolerating the presence of 1% and 3% sodium chloride concentrations in the growth medium . No culture grew in the presence of 1% glycine and 5% NaCl and no culture produced hydrogen monosulphide . All showed good growth in media with carbohydrates which, however, were not attacked . With respect to these characteristics and to literature data, the microbes were classified as Vibro coli, whose isolation had been reported by some authors in relation with dysentery in pigs (Roberts, 1956; Deas, 1960; Lussier, 1962). J Med Primatol, 1976, 5(3), 186 - 94 Malnutrition and susceptibility to infection with Vibrio cholerae in vervet monkeys (Cercopithecus aethiops) . 1 . Induction of protein, B-vitamin complex and calorie malnutrition; Felsenfeld O et al.; Five groups of eight vervet monkeys (Cercopithecus aethiops) were fed regular monkey chow, low protein (LP), low B-vitamin complex (LBV), low protein - low B-vitamin complex (LB-LBV), or protein - low calorie (LP-LC) diet, respectively, for eight weeks . Hematocrit values, hemoglobin, BUN, total protein, albumin, riboflavin and body weight decreased, the ratio of dispensable/indispensable amino acids increased in all animals receiving a deficient diet . Total globulin, IgG and IgM remained unchanged in vervets on LBV diet; panthotenic acid and cyanocobalamin in animals receiving LBV, LP or LP-LC chow; and pyridoxal on the LP or LP-LC diet. Scand J Immunol, 1976, 5(6-7), 867 - 71 Neutralizing antibodies against Escherichia coli and Vibrio cholerae enterotoxins in human milk from a developing country; Holmgren J et al.; By means of the adrenal cell assay and the rabbit small-bowel loop technique enterotoxin-neutralizing antibodies were demonstrated in all investigated milk samples from severely undernourished Pakistani women but, with a single exception, not in milk from Swedish mothers . The antibodies belonged to the IgA and IgG classes as observed in an enzyme-linked immunosorbent assay, which also revealed secretory component on specific enterotoxin antibodies, showing that the IgA antibodies were primarily of the secretory type . The epidemiological stituation and experimental data strongly indicate that Escherichia coli enterotoxin rather than immunologically cross-reactive Vibrio cholerae toxin in most cases constituted the antigenic stimulus for the milk antibodies . The presence in the milk of the Pakistani mothers of secretory IgA antibodies against an important diarrheogenic agent may have practical importance for protection of the breast-fed baby and probably also illustrates the frequent intestinal exposure of these women to enterotoxinogenic bacteria . Since enterotoxin antigen is presumed not to enter deeper tissues, the demonstration of secretory antibodies in milk agrees with a proposed homing of intestinally triggered lymphocytes to the mammary gland. Dev Biol Stand, 1976, 33, 108 - 12 Efficacy of a live oral cholera vaccine in human volunteers; Woodward WE et al.; A live attenuated oral cholera vaccine has been given to volunteers with complete safety . The vaccine strain appeared to multiply significantly in intestinal contents and to result in appreciable vibriocidal antibody response . Subsequent challenge with virulent Inaba vibrios demonstrated substantial protection from clinical illness and from vibrio shedding . The possibility of reversion to toxin production is discussed. Adv Exp Med Biol, 1976, 66, 107 - 15 Cellular events involved in the true primary immune response of splenocytes in vitro; Friedman H et al.; An antibody response showing characteristics of a "true" primary response was readily induced in vitro with splenocytes from normal non-immunized animals cultured with strongly immunogenic Vibrio cholerae somatic antigens . Prior studies have shown that the response to vibrios in intact animals appeared to be a true primary response since no pre-existing antibody forming cells were present in non-immunized animals and the antigen induced response, following a lag of two days, resulted thereafter in a rapid stepwise increase in the number of specific PFCs, reaching a peak at 12 to 14 days . Using the Marbrook culture system for antibody formation a readily detected immunocyte response to vibrios was induced with splenocytes from normal non-immunized animals . No background antibody forming cells developed to the organisms without addition of vibrios in vitro . After in vitro immunization with 10(6) bacteria significant numbers of IgM PFCs appeared with a peak response on days 7 to 8 . Splenocyte cultures from mice primed earlier with vibrios developed a marked secondary response, with appearance of both IgM and IgG PFCs . Large numbers of both classes of PFCs developed, with peak responses on days 6 to 7, similar to the "primary" response to sheep erythrocytes . However, significant numbers of PFCs to vibrios developed in cultures of vibrio-primed cultures even in the absence of vibrios during the 12 day culture period . The availability of a completely in vitro model system to induce a true primary immune response without the complication of pre-existing background antibody forming cells will be of value for further studies concerning various cellular pathways and interactions during the immune response to small amounts of strongly immunogenic bacterial antigens. Arch Virol, 1976, 51(4), 275 - 83 Surface antigens on HeLa cells persistently infected with HVJ (Sendai virus); Kimura Y et al.; Surface antigens of HeLaHVJ cells, a cell line persistently infected with HVJ, were studied by fluorescent antibody staining . After absorption with concentrated HVJ virions and HeLa cells, anti-HeLaHVJ antiserum was able to demonstrate specific surface fluorescence on HeLaHVJ cells, while this serum no longer reacted with original HeLa cells nor with HVJ virions . During cytolytic infection of HeLa cells with HVJ, this specific surface antigen appeared at an early stage of infection prior to the appearance of newly synthesized HVJ viral antigens and moreover appeared in spite of the inhibition of viral protein synthesis . This antigen was detected neither on HeLa cells infected with other myxoviruses except HVJ nor on various other kinds of cells infected with HVJ . The specific surface antigen was still found on the HeLaHVJ cell surface after incubation at 38 degrees C for two days, while HVJ structural antigens on the cell surface no longer could be detected . Mild short-term treatment of HeLa cells with trypsin, neuraminidase from vibrio cholerae, phospholipase-C and hyaluronidase failed to expose specific antigen . The antigen was distinguishable from the Forssman and human blood type antigens . The mechanism of appearance of a new antigen on the surface of HeLaHVJ cells remains unclear. J Supramol Struct, 1976, 4(1), 99 - 120 Structure and function of cholera toxin and hormone receptors; Bennett V et al.; The enterotoxin from Vibrio cholerae is a protein of 100,000 mol wt which stimulates adenylate cyclase activity ubiquitously . The binding of biologically active 125I-labeled choleragen to cell membranes is of extraordinary affinity and specificity . The binding may be restricted to membrane-bound ganglioside GM1 . This ganglioside can be inserted into membranes from exogenous sources, and the increased toxin binding in such cells can be reflected by an increased sensitivity to the biological effects of the toxin . Features of the toxin-activated adenylate cyclase, including conversion of the enzyne to a GTP-sensitive state, and the increased sensitivity of activation by hormones, suggest analogies between the basic mechanism of action of choleragen and the events following binding of hormones to their receptors . The action of the toxin is probably not mediated through intermediary cytoplasmic events, suggesting that its effects are entirely due to processes involving the plasma membrane . The kinetics of activation of adenylate cyclase in erythrocytes from various species as well as in rat adipocytes suggest a direct interaction between toxin and the cyclase enzyme which is difficult to reconcile with catalytic mechanisms of adenylate cyclase activation . Direct evidence for this can be obtained from the comigration of toxin radioactivity with adenylate cyclase activity when toxin-activated membranes are dissolved in detergents and chromatographed on gel filtration columns . Agarose derivatives containing the "active" subunit of the toxin can specifically absorb adenylate cyclase activity, and specific antibodies against the choleragen can be used for selective immunoprecipitation of adenylate cyclase activity from detergent-solubilized preparations of activated membranes . It is proposed that toxin action involves the initial formation of an inactive toxin-ganglioside complex which subsequently migrates and is somehow transformed into an active species which involves relocation within the two-dimensional structure of the membrane with direct perturbation of adenylate cyclase molecules (virtually irreversibly) . These studies suggest new insights into the normal mechanisms by which hormone receptors modify membrane functions. Ann N Y Acad Sci, 1976, 276, 45 - 60 Experimental cancer immunotherapy: modification of tumor cells to increase immunogenicity; Rios A et al.; Firmly established transplantable C3H/HeJ mammary carcinomas can be inhibited by host challenge with Vibrio cholerae neuraminidase (VCN)-treated tumor cells . The effect is totally immunospecific, even VCN-treated tumors bearing shared mammary tumor virus (MTV) antigen cannot induce the regression . Thus, VCN is capable of increasing the immunogenicity of the private, unique-unshared tumor antigens on mammary carcinomas; VCN is incapable of increasing the immunogenicity of the shared MTV-associated tumor antigen even in syngeneic C3HeB/FeJ MTV-free mice . The immunoregressive effect of VCN-treated tumor cells can be augmented by subtotal or total surgical excision of large transplantable tumors . Spontaneous mammary tumors in retired breeder C3H/HeJ female mice can be made to regress by two immunological maneuvers: (1) repeated intratumor injections of VCN and/or BCG; and (2) total excision and immunotherapy with VCN-treated autochthonous mammary tumor cells . The use of VCN-treated transplantable mammary tumor cells sharing the MTV-associated antigen was not better than excision alone . The evidence supports the idea that active specific immunotherapy of spontaneous tumors with VCN-altered tumor cells may require the use of autochthonous cells. Acta Microbiol Pol, 1976, 25(2), 117 - 22 Amino acid metabolism and protein synthesis in streptomycin resistant Vibrio El Tor; Bhattacharya G et al.; Metabolic activities in relation to protein synthesis and amino acid utilization are altered in Vibrio El Tor after development of resistance towards streptomycin . Efficiency of in vivo and in vitro protein synthesis is markedly reduced in streptomycin resistant Vibrio El Tor . The rate of incorporation of 14C-amino acids into protein, uptake of 14C-valine and oxidation of certain amino acids are also altered. Zentralbl Bakteriol {Orig A}, 1975 Dec, 233(4), 531 - 5 Further studies on the behavior of 2-mercaptoethanol resistant antibody following cholera vaccinations in a nonendemic country; Felgner P et al.; Additional data are presented in support of the earlier observation that in nonendemic countries 2-mercaptoethanol (2-ME) resistant vibriocidal antibodies better reflect antibacterial immunity against cholera following vaccination than does total vibriocidal activity . It can be shown that in more than half of the cases even after at least three subsequent vaccinations in six to eight months intervals 2-ME resistant vibriocidins would not last as long as six months . Following intradermal inocculations of vaccine only one in a group of five demonstrated faint 2-ME resistant antibody response . All sera were tested with and without dialysis following 2-ME treatment revealing corresponding results. Zentralbl Bakteriol {Orig B}, 1975 Dec, 161(3), 288 - 94 {Surveys on the occurrence of Vibrio parahaemolyticus on fish caught in the Southern Central Baltic Sea in comparison with other European results (author's transl)}; Zaleski S et al.; Presence of Vibrio parahaemolyticus in European marine environments, including Western Baltic Sea, was ascertained by several scientists . -Our surveys had in view to find out whether fish of the Southern Central Baltic Sea area was contaminated with this microorganism . -In the periods of June - September 1973 and June 1974, 541 fish of 7 species had been tested . Two different technics of isolation were applied . From among 820 colonies picked up for further identification none happened to be Vibrio parahaemolyticus. Endokrinologie, 1975 Dec, 66(3), 279 - 84 Selective chemical changes of gonadotropic hormones by means of neuraminidase from Vibrio cholerae; Finne E; Experiments indicate that Neuraminidase from Vibrio Cholerae selectively cleaves sialic acid derivate from FSH, but leaves the LH sialic acid derivative . By performing a specific test for LH - the OAAD test - we found that the LH activity is not influenced by the presence of Neuraminidase . The FSH activity is destroyed by Neuraminidase but the enzyme itself influences the ovarian weight, resulting in a weight decrease observed 48 hours after injection . Before or after this "critical time" no decrease is observed. J Hyg (Lond), 1975 Dec, 75(3), 399 - 406 Type-specific action of vibriocidal antibody on Vibrio cholerae; Pykett AH et al.; The vibriocidal activity of Inaba and Ogawa antisera, from which cross-reacting agglutinin had been absorbed, was specific for Vibrio cholerae strains of the homologous serotype . No vibriocidal action against strains of the heterologous type was detected . The sera appeared to be equally effective in killing organisms of different biotypes (classical, intermediate, and EITor), provided that these were of the homologous serotype (Inaba or Ogawa) . However, they had been raised against strains of the classical biotype only; and sera resulting from immunization with other biotypes had not been prepared . The implications of these findings in immunity to cholera are discussed. Zentralbl Bakteriol {Orig A}, 1975 Dec, 233(4), 536 - 41 Cholericidic effect of some intestinal disinfectants; Vitez I et al.; Several compounds have been found which are markedly more effective and more economical as chemoprophylactic agents than the tetracycline compounds traditionally used in cholera-endemic areas . Some synergetically acting combinations were shown to exert by several orders of magnitude higher vibriocidal potency than the sum of potency of the compounds when applied singly . In the presence of human intestinal content, however, markedly higher concentrations are needed to exert the proper vibriocidic effect than in aqueous medium . It has been established in guinea pig experiments that the synergistic combinations administered per os in powder form, are able to kill the cholera vibrios within 5 to 10 minutes . The introduction of the appropriate synergistic combinations for controlling the cholera in the endemic areas of Asia and Africa is suggested. Infect Immun, 1975 Dec, 12(6), 1331 - 40 Antitoxic immunity in experimental cholera: protection, and serum and local antibody responses in rabbits after enteral and parenteral immunization; Holmgren J et al.; The protective effect of enternal and parenteral immunization with cholera toxin antigen against experimental cholera in rabbits was studied by using the small-bowel loop technique . Subcutaneous injection of crude toxin as well as purified toxin or toxoids gave rise to significant protection against toxin challenge . The enhanced resistance to toxin was found to correspond to a many-fold higher magnitude of protection against challenge with live vibrios . In the primary response the protection increased during the first month . Booster immunization gave rise to a further increased immunity which, however, declined rapidly . Multiple oral or repeated intraintestinal antigen administrations also induced protective antitoxic immunity although of less magnitude than that obtained by parenteral immunization . Enteral and, to a lesser extent, parenteral immunization gave rise to increased antitoxic antibody titers and immunoglobulin levels in intestinal washings and mucosa scraping . Immunoglobulin G (IgG) and IgG antitoxins predominated, but after enteral immunization total IgA and specific IgA antibodies occasionally reached levels similar to those for IgG . In serum, significantly increased antibody levels (IgG) were only recorded after parenteral immunization . Both the primary binding and the neutralizing antitoxin titers showed a stayistically significant correlation with the degree of protection against toxin challenge; however, for the neutralizing antibodies this correlation was not without exceptions . No relation to protection was found for intestinal antibodies . The results of the present study indicate that enternal as well as parenteral immunization with toxin antigen can give rise to effective cholera immunity . After enternal immunization, the protection appears to be medicated by locally synthesized antibodies . After parenteral vaccination both serum-derived and locally produced antibodies seem to be effective. Biken J, 1975 Dec, 18(4), 187 - 92 Interaction of thermostable direct hemolysin of Vibrio parahaemolyticus with human erythrocytes; Sakurai J et al.; The interaction between thermostable direct hemolysin produced by Vibrio parahaemolyticus WP-1 and human erythrocytes was studied . The lysis of human erythrocytes by the hemolysin was dependent of temperature and no hemolysis occurred at low temperature (0-4 C), but the hemolysin was adsorbed on human erythrocytes even at low temperature . No hemolysis was observed when antihemolysin antiserum was mixed with the hemolysin and human erythrocytes at zero time . On the other hand, lysis of the cells by hemolysin was not completely inhibited when the antiserum was added during the lag time and the inhibitory effect decreased with delay in the time of addition of antiserum . The inhibitory effect of the antiserum decreased with increase in the incubation temperature, increase in the concentration of divalent cations, and decrease in pH . These results suggest that lysis of human erythrocytes by the hemolysin is at least a two-step process consisting of adsorption of the hemolysin to human erythrocytes and the step(s) following adsorption. J Membr Biol, 1975 Nov 7, 24(2), 107 - 29 Mechanism of activation of adenylate cyclase by Vibrio cholerae enterotoxin . Relations to the mode of activation by hormones; Bennett V et al.; The influence of Vibrio cholerae enterotoxin (choleragen) on the response of adenylate cyclase to hormones and GTP, and on the binding of 125I-labeled glucagon to membranes, has been examined primarily in rat adipocytes, but also in guinea pig ileal mucosa and rat liver . Incubation of fat cells with choleragen converts adenylate cyclase to a GTP-responsive state; (-)-isoproterenol has a similar effect when added directly to membranes . Choleragen also increases by two- to fivefold the apparent affinity of (-)-isoproterenol, ACTH, glucagon, and vasoactive intestinal polypeptide for the activation of adenylate cyclase . This effect on vasoactive intestinal polypeptide action is also seen with the enzyme of guinea pig ileal mucosa; the toxin-induced sensitivity to VIP may be relevant in the pathogenesis of cholera diarrhea . The apparent affinity of binding of 125I-labeled glucagon is increased about 1.5- to twofold in choleragen-treated liver and fat cell membranes . The effects of choleragen on the response of adenylate cyclase to hormones are independent of protein synthesis, and they are not simply a consequence to protracted stimulation of the enzyme in vivo or during preparation of the membranes . Activation of cyclase in rat erythrocytes by choleragen is not impaired by agents which disrupt microtubules or microfilaments, and it is still observed in cultured fibroblasts after completely suppressing protein synthesis with diphtheria toxin . Choleragen does not interact directly with hormone receptor sites . Simple occupation of the choleragen binding sites with the analog, choleragenoid, does not lead to any of the biological effects of the toxin. J Reprod Fertil, 1975 Nov, 45(2), 315 - 22 Abortifacient effects of Vibrio cholerae exo-enterotoxin and endotoxin in mice; Gasic GJ et al.; PIP: Examination of the effects of exo-enterotoxin and endotoxin from Vibrio cholerae following intravenous injection into mice at different stages of gestation . Although the patterns of abortificient activity were different, both substances induced termination of pregnancy . When administered between gestation days 4 and 10, exotoxin terminated pregnancy . However, animals more than 10 days pregnant exhibited reducted abortificient activity . Endotoxin, when injected after mid-gestation and when the active principle was heat-stable, was the most effective pregnancy terminator . Exogenous progesterone did not protect the pregnancies nor was it able to prevent the endotoxic effects . Premature parturition was induced in 1/2 to 2/3 of the mice receiving endotoxin on gestation Day 17 while indomethacin reduced the incidence of premature littering . The study indicates that exoand endotoxins exhibit antifertility properties . Rather than inducing progestin deficiency, both appear to act on intrauterine targets . Infect Immun, 1975 Nov, 12(5), 964 - 8 Selective inhibition of cholera toxin- and catecholamine-stimulated lipolysis by blocking agents; Katz MS et al.; Vibrio cholerae enterotoxin stimulates lipolysis in rat epididymal fat cell suspensions . Like hormones this toxin increases adenylate cyclase activity, raising levels of cyclic adenosine 3',5'-monophosphate (cAMP), which activates a cellular lipase . Using specific blocking agents, we studied the responses to the adrenergic lipolytic hormones epinephrine, norepinephrine, and isoproterenol, and to cholera toxin . All stimulators were used at 100 x threshold dose . Propranolol (34 muM), a beta blocking agent, inhibited epinephrine stimulation (P less than 0.001) but not that of toxin (P greater than 0.2) . Choleragenoid (25 mug/ml), a natural toxoid of cholera toxin, blocked stimulation by toxin (P less than 0.001) but not that of the adrenergic agents (P greater than 0.2) . A beta blocker, practolol (3 mM), inhibited stimulation by the catecholamines tested (P less than 0.005) but not that of toxin (P greater than 0.05) . Higher concentrations of propranolol (340 muM) and the alpha blocking agents phenoxybenzamine (3 mM) and phentolamine (1.6 mM) inhibited all agonists (P less than 0.001) . The response to theophylline was inhibited by all blockers (P less than 0.05) except propranolol at the lower concentration (34 muM) . A combined beta and alpha blockade using propranolol and epinephrine together did not inhibit toxin-mediated lipolysis . It appears that stimulation by cholera toxin is independent of beta adrenergic receptors . A major inhibition of theophylline-mediated lipolysis by alpha blocking drugs indicated a nonspecific effect of these agents at the concentrations used . The uninhibited response to toxin in the presence of propranolol and epinephrine suggests a lack of relationship of the toxin receptor to either alpha or beta receptors. J Infect Dis, 1975 Nov, 132(5), 555 - 60 Cytotoxicity of Vibrio parahaemolyticus in HeLa cell culture; Carruthers MM; In an investigation of the effect of Vibrio parahaemolyticus on epithelial cells in vitro, HeLa cells in slide culture chambers were inoculated with washed bacteria and several bacterial preparations . Preparations tested included live and heat-killed whole cells, unconcentrated culture filtrates, whole-cell lysates and their supernatant fluids, and supernatant fluids taken from HeLa cell monolayers previously inoculated with live bacteria . Slide cultures were evaluated at 3 and 5 hr by light microscopy and, in a few instances, by electron microscopy (live preparations) or at 24 hr by light microscopy (nonviable preparations) . Live V . parahaemolyticus had a rapid cytotoxic effect that did not appear to be due to cellular invasion . The nonviable preparations had no effect on HeLa cells . The results demonstrate that V . parahaemolyticus is rapidly cytotoxic to epithelial cells in vitro and suggest that the presence of live organisms may be required for its enteropathogenicity. Health Lab Sci, 1975 Oct, 12(4), 316 - 20 Microorganisms in heated nebulizers; Spaepen MS et al.; Out of 353 heated nebulizers in actual use which were cultured in the Peter Bent Brigham Hospital, 118 (33%) were found to be contaminated . Gram-negative organisms predominated with rare micrococci and fungi . A slow-growing Vibrio was also recovered from 4 nebulizers . Heating of nebulizers above 46 C resulted in significant reduction of contamination . Types of heated nebulizers also figured significantly in the rate of contamination found . Carefully considered recommendations for the care and use of respiratory therapy equipment must be instituted and enforced . Techniques for terminal disinfection of equipment, followed by sterilization should result in the issue of sterile equipment for each patient.Rules for maintenance of equipment at the bedside are also needed. Zh Mikrobiol Epidemiol Immunobiol, 1975 Oct, (10), 98 - 101 {Utilization of serological methods in the retrospective diagnosis of cholera and in detecting vibrion carriers}; Bunin KV et al.; Sensitivity and specificity of the three serological methods were studied comparatively: the vibriocidal test, the reaction of bacterial agglutination and of indirect hemagglutination, with the use of erythrocytes sensitized with the vibrio lyzate, cholera species O-antigen and cholerogen . Investigations were conducted with the blood sera of cholera patients, vibrio carriers and contacts . Vibriocidal test proved to be the most sensitive; its data correlated with the results of bacterial agglutination and indirect hemagglutination with erythrocytes, sensitized with the lysate of the vibrios and the cholera O-antigen . None of the used serological methods provided a 100% coincidence with the results of bacteriological analysis . The frequency of detection of anticholera antibodies decreased in the following order: cholera patients, vibrio carriers, contacts. Infect Immun, 1975 Oct, 12(4), 931 - 3 Inhibition of hemolytic activity of the thermostable direct hemolysin of Vibrio parahaemolyticus by ganglioside; Takeda R et al.; The hemolytic activity of the thermostable direct hemolysin produced by Vibrio parahaemolyticus was inhibited by a ganglioside mixture . The ganglioside component which inhibited the hemolysin was not FM1 ganglioside. S Afr Med J, 1975 Sep 27, 49(41), 1699 - 702 Practical aspects of a cholera surveillance programme; Isaacson M; The recent invasion of southern Africa by cholera stimulated the establishment of a regional cholera reference laboratory and surveillance centre . This article describes its activities in general and details the methods employed in the isolation of Vibrio cholerae from sewage, water, seafoods, patients, and carriers . Importance is also attached to the isolation of NAG (non-agglutinable) V . cholerae and V . parahaemolyticus from certain categories of specimens . In recognition of the importance of regional co-operation in combating the problem of cholera, the collaboration of other countries on the subcontinent was invited, and the response is encouraging. JAMA, 1975 Sep 15, 233(11), 1173 - 6 Tissue invasion by unnamed marine vibrios; Fernandez CR et al.; Vibrio parahaemolyticus is an extremely common organism of major importance as a cause of gastroenteritis, but not common as a cause of tissue infection . Of three patients who had serious localized tissue infections, one died because of an unnamed marine Vibrio infection . Vibrios are easily isolated, although their final speciation is quite difficult in view of their unsettled taxonomic position . It is hoped that agreement can be reached as to bacteriologic genealogy; perhaps then the specific pathogenic manifestations will be clarified . The clinician should consider the possibility of this pathogen in any patient with a wound related to saltwater or seafood. Ann Sclavo, 1975 SEP-OCT, 17(5), 740 - 8 {Characteristics of various NAG vibriones isolated in Italy}; Gatti M et al.; In this paper are reported the results of a comparative study of 4 "NAG" vibrio strains isolated in Italy and the vibrio biotype "E1 Tor" responsible of the 1973 outbreak . The "NAG" vibrios were practically identical with the "E1 Tor" vibrio in biochemical properties, polypeptide composition, enteritogenic activity in rabbit ileal loops and showed various antigenic similarities in gel precipitation and indirect immunofluorescence tests . The possible pathogenic role of the "NAG" vibrios is briefly discussed. Vopr Pitan, 1975 Sep-Oct, (5), 74 - 9 {Experimental study of the action of some physical and chemical factors on the survival and propagation of Vibrio parahaemolyticus in food products}; Ermolina EP et al.; The paper presents data on the ability of Vibrio parahaemolyticus to survive and multiply under different conditions that may be encountered during storage and culinary treatment of food products . Investigations were effected in a nutrient medium and in alimentary products in which the survival terms of Vibrio parahaemolyticus . IV . Occurrence of Vibrio parahaemolyticus in sea mud and sea fish in Hawaii . - secure an almost complete suppression of the Vibrio parahaemolyticus viability . Af greatest epidemiological danger is secondary contamination of food products with Vibrio parahaemolyticus. Zh Mikrobiol Epidemiol Immunobiol, 1975 Sep, 0(9), 29 - 32 {Differences in certain immunological indices among persons who have had cholera and among Vibrio carriers}; Bunin KV et al.; Blood sera of persons, who sustained cholera of different degree of severity, and of vibrio-carriers, and also chromatographic fractions obtained in separation of the mentioned sera on DEAE-cellulose were investigated in the reaction of agglutination and the indirect hemagglutination test . Immunological response in the vibrio carriers was realized by the microglobulin, and in the convalescents--by the macroglobulin type . There was also revealed an association between the severity of the course of cholera and the prevalence in the blood serum of one or another type of specific antibodies: in severe forms of the disease the antibodies of the IgM-class were detected with the greatest frequency, and microglobulin antibodies--in vibrio carriers . The detected differences in the immunological status of those who sustained cholera and of vibrio-carriers could be used as an additional differential sign in the diagnosis of the mentioned conditions. Zh Mikrobiol Epidemiol Immunobiol, 1975 Sep, 0(9), 20 - 5 {Epidemiological patterns of the import and spread of El Tor cholera}; Pavlov AV; The paper is a response to the paper by A . K . Akiev published in 1974 in the "Journal of Microbiology Epidemiology and Immunobiology": "Concerning the Epidemiology of El Tor cholera Abroad" . The opinion of the author concerning the origin of El Tor infection in 1970, the sources of infection, and the factors of its transmission is critisized . Literature data and personal observations explaining the regularities of importation and spread of El Tor cholera as an intestinal infection are presented; these data are against the view of Akiev on El Tor cholera as a disease with a natural nidality caused by freely living vibrios. Isr J Med Sci, 1975 Sep, 11(9), 889 - 95 Viability of Vibrio cholerae biotype El Tor and of cholera phage on vegetables; Gerichter CB et al.; Different vegetables were contaminated with sewage water containing large amounts of El Tor vibrios, and the viability of these vibrios was studied under defined conditions of storage . During the dry season (summer) in Jerusalem, the El Tor vibrios survived for up to 24 hr on parsley, 24 to 30 hr on tomatoes and carrots, 24 to 48 hr on cucumbers, peppers and okra and two to three days on lettuce, when the vegetables were stored at room temperature . The survival time was longer during the rainy season or during storage of the vegetables in a refrigerator . It was also prolonged on damaged vegetables . The El Tor vibrios survived for only a few hours on vegetables exposed to sunlight . The El Tor vibrios survived for 12 to 24 days in experimentally contaminated sewage water, and for up to 10 days in sewage-contaminated soil . The El Tor bacteriophage persisted for up to 36 days on phage-contaminated vegetables. Infect Immun, 1975 Sep, 12(3), 449 - 54 Characterization of the temperature-dependent inactivating factor of the thermostable direct hemolysin in Vibrio parahaemolyticus; Takeda Y et al.; The termostable direct hemolysin of Vibrio parahaemolyticus is inactivated by heating at around 55 C with an inactivating factor isolated from culture filtrates of V . parahaemolyticus . The characteristics of the temperature-dependent inactivating factor were studied by using polyacrylamide gel disc electrophoresis . It was found that when heated with the hemolysin, the inactivating factor destroyed the hemolysin and thus inactivated the hemolytic activity, suggesting that the inactivating factor has proteolytic activity . It was also demonstrated that the inactivating factor itself is heat labile, losing its activity on heating with the hemolysin at 95 C for 15 min . The inactivating factor was stimulated by the presence of NaCl or MgCl2 and showed maximal activity at around pH 8.0 . The results support our previous hypothesis that the Arrhenius effect observed with crude hemolysin of V . parahaemolyticus is due to the presence of a temperature-dependent inactivating factor . The fact that the factor is activated on heating at 50 to 60 C but is inactivated on heating at 70 to 100 C explains the Arrheius effect of crude hemolysin of V . parahaemolyticus. Zh Mikrobiol Epidemiol Immunobiol, 1975 Sep, 0(9), 32 - 6 {Correlation of the antibody concentrations in the cells, subcellular structures and blood serum of rabbits immunized with cholera vaccine}; Adamov AK et al.; Experiments were conducted on animals . Following immunization with cholera vaccine there were revealed vibriocidal antibodies, sometimes -- agglutinating, but no vibriolysins in the cells of the mucosal epithelium of the small intestine, of the liver, the spleen and in the fractions obtained from the cells with the aid of differential centrifugation . This corresponds to the high titre of vibriocidal antibodies in the serum and a relatively low titre of the other types of antibodies . In parenteral immunization with various doses of cholera vaccine the concentration of vibriocidal antibodies reached 10(1)-10(7) per 1 mg of protein, which was millions of times less than in the blood serum. Biull Eksp Biol Med, 1975 Sep, 80(9), 56 - 9 {Isolation under the influence of tetracycline of L-forms of Menchikov's vibrio and the NAG vibrio and their biological properties}; Timakov VD et al.; Stable L-forms were for the first time obtained under the effect of tetracycline on the Mec thnikov vibrio and NAG vibrio . The process of conversion of the L-forms under the effect of tetracycline was the same as under the action of penicilline . Three types of the L-forms were obtained: stable tetracycline-resistant L-forms, stable tetracyclin-superresistant and tetracycline-dependent L-forms . Under the reversion of the unstable L-forms strains with a typical complex of biological properties and atypical revertants were obtained. S Afr Med J, 1975 Aug 30, 49(37), 1514 - 6 Vibrio parahaemolyticus -- a marine pathogen detected in South African coastal waters; Bubb HD; Vibrio parahaemolyticus has been detected in the Natal and Eastern Cape costal water of South Africa . As a proven cause of gastro-enteritis or a potential pathogen to humans, it is an organism of importance to public health . Outlines of the history, pathogenicity and identification are given, as well as the results of the local investigation of this organism. Arch Microbiol, 1975 Aug 28, 104(3), 285 - 8 Effect of temperature, salts, pH, and other factors on the development of peritrichous flagella in Vibrio alginolyticus; Ulitzur S; The development of peritrichous flagella and, consequently, swarming of Vibrio alginolyticus depend on a complex relationship between temperature, salt concentrations and pH . At temperatures above 28 degrees C V . alginolyticus did not develop peritrichous flagella unless certain minimal concentrations of NaCl are present: the higher the temperature, the higher the NaCl concentrations required for peritrichous flagella synthesis . This requirement for NaCl at high temperatures is much more pronounced at pH 9 than at pH 6 . High temperatures and low concentrations of NaCl also inhibited swarming of cells already armed with peritrichous flagella . Other cations, such as Li+, K+ and Mg2+ . replaced NaCl only at temperatures below 28 degrees C. Jpn J Med Sci Biol, 1975 Aug, 28(4), 215 - 21 Serotypes of Vibrio parahaemolyticus from clinical and environmental sources in Togo (West Africa); Bockmuhl J et al.; Serological analysis of O and K antigens was performed on 343 strains of Vibro parahaemolyticus isolated from clinical and environmental sources in Togo . Only two strains were not typable by the available O antisera . K untypable strains were found in 4.8% of isolates from gastroenteritis patients, in 11% from healthy carriers, and in 47% and 46% of isolates, respectively, from water and fish samples . Thirteen serotypes identified in Togo are not considered in the Japanese antigenic scheme . The suitability of the Japanese typing scheme for geographic areas outside of Japan is discussed and its extension is suggested. Can J Microbiol, 1975 Aug, 21(8), 1272 - 4 Characterization of the factors responsible for death of fish infected with Vibrio anguillarum; Umbreit TH et al.; Vibrio anguillarum produced substances toxic for goldfish (Crassius auretus) that are released from living bacteria and associated with heat-killed bacteria . Heating (100 degrees C) enhances the potency of the extracellular toxin. Appl Microbiol, 1975 Aug, 30(2), 251 - 7 Incidence of Vibrio parahaemolyticus in Chesapeake Bay; Kaneko T et al.; A Bay-wide survey of the distribution of Vibrio parahaemolyticus was carried out in Chesapeake Bay during May 1972, to determine whether the annual cycle of V . parahaemolyticus which was observed to occur in the Rhode River subestuary of Chesapeake Bay took place in other parts of Chesapeake Bay . In an earlier study, April to early June, when the water temperature rises from 14 to 19 C, was found to be a critical period in the annual cycle of the organism in the Rhode River, since this is the time period when the annual cycle is initiated . Results of this study, however, revealed that V . parahaemolyticus could not be found in the water column during May 1972 . Nevertheless, several samples of sediment and plankton yielded V . parahaemolyticus isolates . Comparison of data with those for the Rhode River area examined in the earlier studies of the annual cycle of V . parahaemolyticus suggests that the time of initiation of the annual cycle of V . parahaemolyticus in the open Bay proper may be influenced by various factors such as temperature and salinity, i.e., deeper water locations may show initiation of the V . parahaemolyticus annual cycle later than shallow areas . Confirmation of the presence of the organisms in the samples studied was accomplished using numerical taxonomy with 19 reference strains also included in the analyses. J Infect Dis, 1975 Aug, 132(2), 181 - 8 Antibody cross-linking as a factor in immunity to cholera in infant mice; Bellamy JE et al.; Several antibody preparations were tested for their ability to reduce adsorption of Vibrio cholerae to isolated intestinal epithelial cells, and this ability was related to agglutination and protective activity in infant mice . The results demonstrate that (1) the reduction in adsorption of V . cholerae to epithelial cells correlates with the degree of agglutination for given antibody preparation; (2) intact tantibodies protect infant mice from cholera only at concentrations that agglutinate the bacteria; and (3) purified antibodies to flagellar antigens protect infant mice from cholera . These results indicate that cross-linking of bacteria by antibody causes a reduction in the number of organisms adsorbed to the intestinal wall . Thus antibody cross-linking plays an important role in immunity to cholera in infant mice. J Infect Dis, 1975 Aug, 132(2), 175 - 80 Further evidence for cross-linking as a protective factor in experimental cholera: properties of antibody fragments; Steele EJ et al.; Enzymic fragments of IgG antibody were tested for their protective abilities in the infant mouse cholera model . F(ab')2 retained the full protective activity of the parent IgG molecule despite losses in complement fixation and opsonic functions . Fab' and Fab fragments also contained significant protective activity but at a level of only 10% of the intact IgG or F(ab')2 . Self-recombinant univalent F(ab')2 also contained about 10% of the protective activity of the divalent F(ab')2 parent molecule . These results are interpreted as evidence that cross-linking is an important mechanism by which specific antibody protects mucosal surfaces against experimental infection with Vibrio cholerae. Lab Invest, 1975 Aug, 33(2), 187 - 92 Bovine venereal vibriosis . Ultrastructure of endometrial inflammatory lesions; Corbeil LB et al.; The inflammatory lesions in bovine venereal vibriosis have been examined by light and transmission electron microscopy in order to provide a better understanding of host defense of the uterus against bacterial invasion . Neutrophils and eosinophils were found mainly at the endometrial surface and in glandular lumina . Mononuclear cells within the endometrial tissue were identified ultrastructurally . Lymphocytes were most abundant, plasma cells were next in frequency, and mononuclear phagocytes were least often observed . An attempt was made to classify lymphocytes on the basis of presence or absence of a cytoplasmic network of filaments and other ultrastructural characteristics. Immunology, 1975 Aug, 29(2), 283 - 99 Vibriolytic IgG immunocyte response of mice after primary and secondary immunization with cholera somatic antigens; Friedman H; Antibody plaque-forming cells (FC) to the somatic antigens of Vibrio cholerae were enumerated in the spleen of mice after primary and secondary immunization with a heat-killed vaccine prepared from the vibrios . Immunocytes releasing both high efficiency IgM and low efficiency IgG antibody were readily detected using a direct and facilitated plaque procedure in agar gel . Whereas the peak numbers of IgM-PFC after primary immunization occurred on days 12 to 14, the peak IgG-PFC response developed somewhat later (16-18 days) . After a second injection of vaccine larger numbers of both IgM- and IgG-PFE appeared in the mouse spleens, with peak responses for both occurring between days 5 and 8 . The largest number of IgG-PFC developed in spleens of mice given a second injection of vaccine 6-8 weeks after primary immunization . The dose of killed vibrios used for priming markedly affected both the magnitude and the class of antibody-forming cells appearing during the secondary response; 1--10 mug vaccine was more effective than higher or lower doses for priming the mice to a heightened secondary response . Furthermore, the antigenic specificity of both the IgM- and IgG-PFC appearing after secondary immunization was directly related to the strain of cholera bacilli used for priming . When mice were immunized with the Ogawa strains of cholera most of the secondary PFC after booster immunization with the serologically distinct Inaba strain was directed towards the common antigen shared by both strains and not to the type specific antigen of the Inaba vibrios . The specificity of the anti-vibrio PFC during both the primary and secondary responses was readily demonstrable by inhibition experiments using sonicated or soluble cholera antigens . Prior incubation of these antigens with test spleen cells in the agar gel effictively inhibited development of the vibriolytic plaques, regardless of antibody class . Similar antigen extracts from toher bacteria had no effect . The immunoglobulin nature of the plaques was also demonstrable by inhibition with low dilutions of rabbit anti-mouse globulin serum incorporated into the agar plates prior to testing; both IgM and IgG plaues were inhibited. Zh Mikrobiol Epidemiol Immunobiol, 1975 Jul, (7), 90 - 2 {Electron microscopy of cholera vibrios from the intestinal contents of infected suckling rabbits}; Lobanov VV et al.; Electron microscopic study of the cells of classic, El Tor and NAG-vibrios showed them to be no different by the structure type from the cells of ofter Gram-negative bacteria . A characteristic peculiarity of the cholera vibrios revealed after their passage through the intestine of nursling rabbits was the presence of microcapsules and protrusions of the areas of the wall membranous apparatus. Zh Mikrobiol Epidemiol Immunobiol, 1975 Jul, (7), 33 - 7 {Use of disc electrophoresis in polyacrylamide gel method for the separation of soluble proteins of vibrios}; Atarova GT et al.; The authors present the results of a comparative study of the protein spectra of 80 strains of various vibrios . Proteinograms were elaborated by the method of disc-electrophoresis in polyacrylamide gel for each strain and species . There were revealed 25 protein peaks with definite mobility coefficients . The number of peaks and their areas varied in different species . The qualitative and the quantitative differences between the vibrio species were established . Results of investigations demonstrated a possibliity of using the protein spectra for the differentiation of microorganisms. Am J Trop Med Hyg, 1975 Jul, 24(4), 638 - 40 The first reported case from Panamá of acute gastroenteritis caused by Vibrio parahaemolyticus; Kourany M et al.; The first confirmed case of Vibrio parahaemolyticus gastroenteritis in Panama is reported . This marine organism, causing food poisoning in some countries and isolated only recently from seawater in Panama, was recovered from the stools of a patient with acute gastroenteritis, as well as from fresh shrimp used in preparing seafood dishes . This person and 39 other individuals became acutely ill a few hours after eating shrimp at the same restaurant. Immunology, 1975 Jul, 29(1), 197 - 208 Dog immunoglobulins . II . The antibacterial properties of dog IgA, IgM and IgG antibodies to Vibrio cholerae; Heddle RJ et al.; Secretory IgA antibodies to Vibrio cholerae were purified from the parotid saliva and mammary secretions of locally and orally immunized dogs using gel filtration, ion-exchange chromatography and anti-immunoglobulin immuno-absorbents . IgM and IgG antibodies were isolated from serum by gel filtration and ion-exchange chromatography . IgA antibodies proved to have minimal, if any, activity in direct killing of bacteria in the presence of complement or in the promotion of phagocytosis . The minimal activity which IgA had in these assays could be accounted for by extremely small quantities of IgM antibody . The same IgA antibodies, mixed with the challenge innoculum of Vibrio cholerae and fed to infant mice, protected these mice as efficiently as IgG or IgM antibodies. Ann Intern Med, 1975 Jul, 83(1), 107 - 10 Health hazards of bivalve-mollusk ingestion; Earampamoorthy S et al.; Bivalve mollusks (oysters, clans, and mussels) filter large quantities of water unselectively and thereby may concentrate a variety of aquatic contaminants pathogenic for man within edible shellfish viscera . The recognized bacterial disease associated with ingestion of contaminated bivalves include typhoid fever (not presently a public health problem), Vibrio parahemolyticus gastroenteritis, and Vibrio chloerae infection . The major known shellfish-associated viral diseases are viral hepatitis and possibly viral gastroenteritis . The ingestion of bivalves that have fed on the toxic species of dinoflagellates that produce red tides may be responsible for an uncommon and very rarely fatal illness, paralytic shellfish poisoning . Outbreaks of airborne respiratory irritation in populations exposed to red tides may be the most common public health problem associated with red tides . The health hazards resulting from industrial, agricultural, and oil pollution of bivalves in coastal waters and the hazard from improper handling of bacterially contaminated mollusks remain to be defined. Proc Natl Acad Sci U S A, 1975 Jul, 72(7), 2520 - 4 Interaction of cholera toxin and membrane GM1 ganglioside of small intestine; Holmgren J et al.; Ganglioside GM1 was isolated from the small intestinal mucosa of man, pig, and beef and amounted to 0.1, 2.0, and 43 nmol per g fresh weight, respectively . These differences in GM1 content were associated with a quantitatively differing ability of the mucosal cells to bind cholera toxin . Human cells bound about 15,000 toxin molecules when saturated with the toxin, porcine cells 120,000, and bovine cells 2,600,000 molecules . The association constant (KA) of the cholera toxin binding was, for cells of all three species, about 10(9) liters/mol . Exogenously added GM1 ganglioside was incorporated in intestinal mucosal cells as well as in intact rabbit small bowel . The increment in GM1 was associated with a correspondingly increased number of binding sites for cholera toxin, whereas KA was unchanged . GM1 incorporation increased the sensitivity of the rabbit small bowel to the diarrheogenic action of cholera toxin . Vibrio cholerae sialidase hydrolyzed isolated intestinal diand trisialogangliosides to GM1 . However, the enzyme did not change the ganglioside pattern of intestinal mucosa, had very little influence on the number of toxin binding sites on intestinal cells, and did not alter the sensitivity of the small bowel to the diarrheogenic action of the toxin . These results demonstrate a relationship in the intestinal mucosa between the GM1 ganglioside concentration, the number of binding sites for cholera toxin, and the sensitivity to the biologic action of the toxin . Thus, the study strongly supports the concept that the GM1 ganglioside is the intestinal binding receptor for cholera toxin. Arch Intern Med, 1975 Jul, 135(7), 994 - 5 Bilateral deep brachial vein thrombophlebitis due to vibrio fetus; Vesely D et al.; A patient had bilateral deep brachial vein thrombophlebitis in which Vibrio fetus was recovered from six blood cultures of the six drawn . Fever and phlebitis continued with treatment with intravenous doses of heparin and oxacillin but rapidly improved with treatment change of oxacillin to clindamycin . In vitro antibiotic disk susceptibility testing confirmed resistance to oxacillin and susceptibility to clindamycin . Vibrio fetus infection is associated with the vascular endothelium in this and previously reported cases Vet Med (Praha), 1975 Jul, 20(7), 407 - 13 {Comparison of nutrient media for isolation of microflora in swine dysentery}; Krizanova H et al.; The usability of five nutrient media - three kinds of spirolate media, thioglycolate medium and brain hearth medium - suitable for the isolation of Vibrio coli and germs similar to borrelia isolated from pigs affected by dysentery, and vibria isolated from cattle, was compared in the study . The results of comparison show that fresh-prepared substrates must be used bor the isolation of the mentioned germs in all cases and a corresponding medium must be provided . The modified spirolate media prepared from inland and imported ingredients were suitable for the isolation of the mentioned microorganisms even in the case of their use as selective media after enrichment with antibiotics . The spirolate media and thioglycolator agar were not found suitable for maintaining the strains of germs similar to borrelia due to the quick production of involution forms and the result dying of the strains . Thenutrient solution prepared as the brain hearth inf . Oxoid blood agar is very suitable for the isolation as well as maintenance of strains. Infect Immun, 1975 Jul, 12(1), 187 - 92 Partial purification and properties of somatic antigen spontaneously released from Vibrio cholerae; Pike RM et al.; The supernatant fluids from cultures of Vibrio cholerae grown for 40 h in an dialyzable medium were dialyzed, concentrated, and fractionated on agarose columns . The fractions containing most of the antigen which inhibited the vibriocidal activity of homologous antiserum were pooled, dialyzed, and concentrated to provide material with about 100 times the specific activity of the original culture supernatant . This material, containing 12 to 20% protein, 11 to 19% carbohydrate, and about 16% unbound lipid, had a mean lethal dose for mice of about 500 mug . This partially purified antigen absorbed all the vibriocidal antibody from homologous antiserum against live cultures and produced a single line of precipitation in gel diffusion tests with the same antiserum. Infect Immun, 1975 Jul, 12(1), 81 - 7 Nature of the Kanagawa phenomenon of Vibrio parahaemolyticus; Chun D et al.; In a study of the Kanagawa phenomenon of Vibrio parahaemolyticus, both Kanagawa-positive and -negative strains were found to produce hemolytic factors that could not be differentiated on Wagatsuma blood agar . The presence of fermentable carbohydrates in media containing high concentrations of NaCl promoted the growth of V . parahaemolyticus and resulted in a marked decrease in medium pH and increased hemolysin production . The Kanagawa hemolysis of test strains differed according to the carbohydrates added . Clearly defined Kanagawa hemolysis was observed in blood agars of high salt content, but the distinction was lost in media containing 3% NaCl . From the results of this study, the Kanagawa hemolysis was interpreted as an expression of quantitative difference in hemolysin production, a conclusion that is clearly demonstrated on special blood agar of high salt content. Lancet, 1975 Jun 28, 1(7922), 1400 - 2 Sucrose in oral therapy for cholera and related diarrhoeas; Nalin DR; PIP: The use of sucrose in oral rehydration therapy solutions in place of glucose was tested in 18 patients, 17 males and 1 female, admitted for treatment of severe dehydration due to diarrhea and vomiting . 13 of these patients were positive for cholera (1 with untyped vibrio), whereas 4 others cultured no recognizable pathogen . Patients received an average 1100 ml of intravenous fluids to keep the intravenous drip open during the oral therapy period, and the intravenous therapy was stopped or slowed during oral (or nasogastric) therapy . Average patient age was 32 years . Oral solutions contained either 48 or 38 gm of sucrose per liter plus (in all solutions) sodium chloride (4.2 gm/liter), sodium bicarbonate (2 gm/liter), and potassium citrate (2.7 gm/liter) . Of the 18 patients, 15 could be maintained using this solution, but 3 developed massive increases in net fluid losses with increases in plasma specific gravity, which necessitated terminating the therapy . In these failure cases, plasma specific gravity increased over 1.031 . Stool samples of 12 patients tested were found to contain reducing sugar: prehydrolysis 436 mg/100 ml, posthydrolysis 957 mg/100 ml . The breakdown of sucrose by intestinal enzymes or by bacteria accounts for the presence of reducing sugar in the stool . These data contrast with the rarity of treatment failures of oral glucose therapy; therefore, glucose is the preferable component in oral rehydration electrolyte solution therapy . Arch Microbiol, 1975 Jun 20, 104(1), 67 - 71 The mechanism of swarming of Vibrio alginolyticus; Ulitzer S; Factors leading to swarming of Vibrio alginolyticus cells on solid media were studied . Polar flagellated rods from liquid medium develop into small colonies on solid medium . Byproducts, accumulating in the colony area, induce at certain critical concentrations, the formation of peritrichous flagella and development of long heavily flagellated filaments which swarm away form the high by-product concentrations . Several types of nonswarming mutants were isolated, among them, mutants which lack the capacity to form swarming-inducing pyproducts, but can be induced to swarm by byproducts of other mutants incapable of swarming . Different compounds could replace the natural metabolic byproducts; at very low concentration these compounds induce peritrichous flagella and swarming in some of the nonswarming mutants mentioned above . The natural metabolic byproducts accumulating in yeast-extract-peptone medium are suggested to be volatile acids belonging to the valine and isoleucine pathway . Wild-type V . alginolyticus cells cannot swarm on certain substrates but its mutants, able to swarm on many substrates in minimal media, are easily selected. J Mol Evol, 1975 Jun 9, 5(1), 25 - 34 Procaryote phylogeny IV: concerning the phylogenetic status of a photosynthetic bacterium; Zablen L et al.; The 16S ribosomal RNA (30S subunit) of Rhodopseudomonas spheroides has been characterized in terms of T1 ribonuclease digestion products . This "fingerprint" ultimately permits the placement of R . spheroides into a detailed procaryotic phylogenetic tree . Given the number of major procaryotic lines that have been characterized in these terms to date, one can tentatively place the Athiorhodaceae closer to the Vibrio-Enteric group than to the Bacillaceae or Cyanophyta. J Membr Biol, 1975 Jun 3, 22(1), 29 - 52 Mechanism of activation of adenylate cyclase by Vibrio cholerae enterotoxin; Bennett V et al.; The kinetics and properties of the activation of adenylate cyclase by cholera enterotoxin have been examined primarily in toad erythrocytes, but also in avian erythrocytes, rat fat cells and cultured melanoma cells . When cholera toxin is incubated with intact cells it stimulates adenylate cyclase activity, as measured in the subsequently isolated plasma membranes, according to a triphasic time course . This consists of a true lag period of about 30 min, followed by a stage of exponentially increasing adenylate cyclase activity which continues for 110 to 130 min, and finally a period of slow activation which may extend as long as 30 hr in cultured melanoma cells . The progressive activation of adenylate cyclase activity by cholera toxin is interrupted by cell lysis; continued incubation of the isolated membranes under nearly identical conditions does not lead to further activation of the enzyme . The delay in the action of the toxin is not grossly dependent of the number of toxin-receptor (GM1 ganglioside) complexes, and is still seen upon adding a second dose of toxin to partially stimulated cells . Direct measurements indicate negligible intracellular levels of biologically active radioiodinated toxin in either a soluble or a nuclear-bound form . The effects are not prevented by Actinomycin D (20 mug/ml), uromycin (30 mug/ml), cycloheximide (30 mug/ml), sodium fluoride (10 mM) or sodium azide (1 mM); KCN, however, almost completely prevents the action of cholera toxin . The action of the toxin is temperature dependent, occurring at very slow or negligible rates below certain critical temperatures, the values of which depend on the specific animal species . Thetransition for toad erythrocytes occurs at 15 to 17 degrees C, while rat adipocytes and turkey erythrocytes demonstrate a discontinuity at 26 to 30 degrees C . The temperature effects are evident during the lag period as well as during the exponential phase of activation . The rate of decay of the stimulated adenylate cyclase activity of cultured melanoma cells indicates a half-time of about 36 hr . The rate of exponentially increasing activity and extent of enzyme activation are related to the number of bound toxin molecules according to a Langmuir adsorption isotherm and are half-maximal when about 2000 molecules of toxin are bound per cell . It is proposed that initially cholera toxin binds ineffectively, but that it is converted to an active form during the lag phase . This process may involve lateral motion of toxin-GM1 ganglioside complex within the plane of the membrane . The kinetics of adenylate cyclase activation are consistent with the possibility that during the exponential phase a bimolecular association is proceeding between the active form of the cholera toxin and some other membrane component . The possibility is considered that the cholera toxin molecule may bind directly to adenylate cyclase . These considerations may prove useful in understanding the possible interactions of active hormone-receptor complexes with adenylate cyclase in cell membranes. J Membr Biol, 1975 Jun 3, 22(1), 1 - 28 Mechanism of action of Vibrio cholerae enterotoxin . Effects on adenylate cyclase of toad and rat erythrocyte plasma membranes; Bennett V et al.; The characteristics of the cholera toxin-stimulated adenylate cyclase of toad (Bufus marinus) and rat erythrocyte plasma membranes have been examined, with special emphasis on the response to purine nucleotides, fluoride, magnesium and catecholamine hormones . Toad erythrocytes briefly exposed to low concentrations of cholera toxin (40,000 to 60,000 molecules per cell) and incubated 2 to 4 hr at 30 degrees C exhibit dramatic alterations in the kinetic and regulatory properties of adenylate cyclase . The approximate Km for ATP, Mg++ increases from about 1.8 to 3.4 mMin the toxin-stimulated enzyme . The stimulation by cholera toxin increases with increasing ATP, Mg++ concentrations, from 20 percent at low levels (0.2 mM) to 500 percent at high concentrations (greater than 3 mM) . Addition of GTP, Mg++ (0.2 mM) restores normal kinetic properties to the toxin-modified enzyme, such that stimulation is most simply explained by an elevation of Vmax . GTP enhances the toxin-treated enzyme activity two- to fourfold at low ATP concentrations, but this effect disappears at high levels of the substrate . At 0.6 mM ATP and 5 mM MgC12 the apparent K alpha for GTP, Mg++ is 5 to 10 muM . The control(unstimulated) enzyme demonstrates a very small response to the guanyl nucleotide, 5'-ITP also stimulates the toxin-treated enzyme but cGMP, guanine, and the pyrimidine nucleotides have no effect . Cholera toxin also alters the activation of adenylate cyclase by free Mg++, decreasing the apparent K alpha from about 25 to 5 mM . (minus)-Epinephrine sensitizes the toad erythrocyte adenylate cyclase to GTP and also decreases the apparent K alpha for free metal . Sodium fluoride, which causes a 70- to 100-fold activation of enzyme activity, has little effect on sensitivity to GTP, and does not change the apparent K alpha for Mg++; moreover,it prevents modulation of these parameters by cholera toxin . Conversely, cholera toxin severely inhibits NaF activation, and in the presence of fluoride ion the usual three to fivefold stimulation by toxin becomes a 30 to 60 percent inhibition of activity . The toxin-stimulated enzyme can be further activated by catecholamines; in the presence of GTP the (minus)-epinephrine stimulation is enhanced by two- to threefold . The increased catecholamine stimulation of toad erythrocyte adenylate cyclase induced by cholera toxin is explained primarily by an increase in the maximal extent of activation by the hormones . Rat erythrocyte adenylate cyclase is also modified by cholera toxin . In the mammalian system the apparent affinity for the hormone appears to be increased . Cholera toxin thus induces profound and nearly permanent changes in adenylate cyclase by a unique process which mimics the stimulation by hormones in important ways, and which also accentuates the normal hormonal response . The relevance of these findings to the mechanism of action of cholera toxin is considered. Trop Geogr Med, 1975 Jun, 27(2), 209 - 10 Vibrio parahaemolyticus as a cause of gastroenteritis in Indonesia; Bonang G et al.; Vibrio parahaemolyticus was isolated from rectal swabs in two cases of acute gastroenteritis from different districts of Jakarta. Zh Mikrobiol Epidemiol Immunobiol, 1975 Jun, (6), 81 - 5 {Pathogenicity of nonagglutinating vibrios}; Dunaev GS et al.; In studying the pathogenicity of nonagglutinating vibrios it was established that the majority of the strains isolated from the patients suffering from enteritis possessed enteropathogenic properties which were revealed in the trials on nursling rabbits and on the isolated intestinal loop a of an adult rabbit . In difference to the cholera vibryos, these microorganisms produced no typical cholerogenicity syndrome expressing, however, a number of enteropathogenic properties: caused diarrhea, overfilling of the intestine with fluid, etc . Autopsy showed a typical enterocolitis picture, confirmed by histological studies . Nonagglutinating vibrio cultures isolated from the water and from healthy persons possessed no enteropathogenic properties . An isolated intestinal loop of an adult rabbit proved to be the most sensitive experimental model. Zh Mikrobiol Epidemiol Immunobiol, 1975 Jun, (6), 78 - 81 {Ultrastructure of L forms . IV . L forms of nonagglutinating vibrios}; Shul'ga MA et al.; A study was made of the ultrastructure of stable L-forms of Nag vibrios aged 24 hours . Cells of all types of the L-forms had cytoplasmic membranes, and a three-layered structure, which was found not everywhere . Externally of the cytoplasmic membrane, in some areas of the individual cells there were revealed a plastic layer of cell wall and a basal membrane . However, in difference to bacterial forms of the vibryos, rigidity of the cell wall was disturbed, and the links between the cell wall and the cytoplasmic membrane were indetectable . There were regularly revealed lamellar of myelin-like membranous structures in the cytoplasm, which did not occur in bacterial forms, and also lamellar mesosomes . The latter were found in the sites of cell division . Viability of small bodies as the minimal reproductive forms of the L-cultures is confirmed by the presence in them of a nucleoid and of the binary division. Gastroenterology, 1975 Jun, 68(6), 1556 - 62 Significance of elevated liver alkaline phosphatase in serum; Brensilver HL et al.; The serum alkaline phosphatase was fractionated by polyacrylamide gel electrophoresis in 317 patients with elevated serum alkaline phosphatase activity . In 253 patients the source of the elevation was the isoenzyme of presumed liver origin, band L . In 87 of these patients, there was either no obvious liver disease or the alkaline phosphatase elevation was inappropriately high . In 19 of the 87, liver disease was further excluded by liver biopsy or by laparotomy . Because of this, biochemical studies were done to verify the hepatic origin of band L . Band L and alkaline phosphatase extracted from human liver migrated together on polyacrylamide gel electrophoresis before and after digestion with Vibrio cholerae neuraminidase . They had identical pH optima, sedimentation coefficients, Michaelis constants, and rates of inactivation at 55.5 degrees C . They had different rates of inactivation in 3 M urea . Over-all, the data indicate that band L is of liver origin, and that elevation of the hepatic alkaline phosphatase isoenzyme may be a nonspecific finding in certain patients. Zh Mikrobiol Epidemiol Immunobiol, 1975 Jun, (6), 48 - 51 {Characteristics of the immunogenic properties of cholerogen-anatoxin according to gel precipitation reactions and quantity of reducing sugars}; Dzhaparidze MN et al.; The authors suggest the use of determination of the titre of the diffuse precipitation in gel test with the O-cholera serum, and the content of reducing sugars after Hagedorn-Jensen to characterize the antibacterial properties of the cholerogen-toxoid preparation along with (or instead of) the active mouse protection (AU50) test used earlier, because a close correlative association was established between them . No correlative association proved to exist between the vibriocidal antibody titre of the sera of rabbits immunized with the cholerogen-toxoid and the mouse protection (AU50) test or the titre of the precipitation in gel test in the preparation. Appl Microbiol, 1975 Jun, 29(6), 814 - 8 Method for detecting small numbers of Vibrio cholerae in very polluted substrates; Sechter I et al.; A method is presented for the indirect detection of Vibrio cholerae by the multiplication of two specific bacteriophages: phiH74/64 for El-Tor vibrios, and phage group IV (Mukerjee) for classical vibrios . The product to be examined is seeded in alkaline tryptone water for enrichment, as in the classical method, and is then incubated for 6 h at 37 C . Thereafter, a loopful is transferred to each of two nutrient broth (pH 9) tubes . One of these receives a drop of phage phiH74/64; the other receives a drop of phage group IV . The stock phages are diluted so as to contain about 3,800 plaque-forming units in one drop; this is the maximum amount which, when added to 10 ml of broth, will not be detected in a loopful of 1 mm diameter . The tubes containing phage phiH74/64 are incubated at 42 C; those with phage group IV are incubated at 37 C . After 18 h the cultures are killed by agitation with chloroform, and a 1-mm loopful is deposited on a layer seeded with the detector strains: Makassar 757 for El-Tor phage and V . cholerae 154 for classical cholera phage . After 4 to 5 h at 37 C, lysis appears on the spot areas if there has been phage multiplication in the respective broth tubes . With experimentally contaminated sewage water, vegetables, or stools, 1 to 10 cholera vibrios were detected in every sample . In rare cases, false-positive results were obtained by multiplication of the phage on non-cholera vibrios. Acta Pathol Microbiol Scand {B}, 1975 Jun, 83(3), 293 - 4 Boric acid tolerant Vibrio cholerae: biological and physical properties; Ojo MO; Two boric acid tolerant variants were developed by passage in boric acid containing media from a mouse virulent parent Vibrio cholerae strain . The variants were stable, exhibited increased acid resistance, were less virulent to mice, but protected against challenge with the mouse virulent parent strain. J Natl Cancer Inst, 1975 Jun, 54(6), 1379 - 84 Augmented uptake of neuraminidase-treated sheep red blood cells: participation of opsonic factors; Schmidtke JR et al.; Neuraminidase from Vibrio cholerae (VCN) was used to treat sheep red blood cells (SRBC) which were then incubated in vitro with murine peritoneal macrophages . The uptake of VCN-treated SRBC by macrophages was greater than the uptake of SRBC not treated with VCN . SRBC opsonized with normal mouse serum (NMS) were taken up to a greater extent than untreated SRBC . SRBC treated with VCN and opsonized with NMS were phagocytosed to a greater extent than untreated SRBC, VCN-treated SRBC, or opsonized SRBC . Evidence demonstrated that factors in serum from normal C3H/HeJ mice augmented the uptake of VCN-treated SRBC in greater amounts than of normal SRBC . These findings were discussed in relation to the increased immunogenicity of neuraminidase-treated cells. Antibiotiki, 1975 Jun, 20(7), 623 - 8 {Analysis of the data in the literature on the sensitivity of cholera vibrios to tetracycline}; Semiotrochev VL et al.; Variation diagrams were proposed to be used for the analysis of the published data on the study of Vibrio cholerae sensitivity to tetracycline . The diagrams provided systematization of dissimilar data of the experimental studies and a unique system of estimation was created . The analysis of the systematized data provided dividing of the Vibrio cholerae strains tested into 4 groups differing in the levels of their sensitivity to tetracycline . The groups were independent of the Vibrio cholerae biotypes and the place and period of the strain isolation. Ann Sclavo, 1975 May-Jun, 17(3), 489 - 99 {Studies on the efficacy of cholera vaccination in a population sample of Bari}; Barbuti S et al.; To measure the efficacy of cholera vaccination during the epidemic of cholera occurred in August-September 1973 in Bari, agglutinating and vibriocidal antibody titers were determined in 151 subjects; of these, 65 were children between 6 months and 4 years of age and 86 adults past thirty years . Antibodies could be proved in 80% of the subjects, with relatively higher frequency in children (84,6%) than in adults (76,7%) . No significant difference was seen against the serotypes Inaba and Ogawa . The vibriocidal antibody tests was uniformly more sensitive than the agglutination test . In 108 of these subjects (all of 65 children and 43 of the adults), it was possible to make the serological controls 3 and 6 months after vaccination . At 3 months, antibodies were determined in 50,7% of the children and in 44,2% of the adults; at 6 months, only 20% of the children and 18,6% of the adults showed agglutinating and vibriocidal antibodies . For the control, the research of V . cholerae-antibodies was maked also in sera of 300 healties subjects of all ages, by whom the bleedings were obtained prior to epidemic of cholera . Antibodies against the serotypes Inaba and Ogawa were determined in 23 subjects (7,6%); of these, 5 had vibriocidal titers of 1:2560-1:5120. Ann Sclavo, 1975 May-Jun, 17(3), 480 - 8 {Studies on the spread of Vibrio cholerae, biotype El Tor in the environment during the period following the 1973 epidemic in Apulia}; Leogrande G et al.; In the period following the cholera epidemic, studies were made of sewage, seawater and various foodstuffs (green vegetables, mussels, etc.) in an attempt to isolate Vibrio cholerae of the El Tor biotype . Only in the samples of sewage taken at Bari and Taranto was it possible to isolate Vibrio cholerae biotype El Tor for a period of about a month after the last case of the illness; in this period it was isolated in 5 of the 11 samples taken at Bari and in 1 of the 14 taken at Taranto . The positive tests involved all zones of the city, leading to the conclusion that the bearers had spread throughout the whole population . The cholera-bearing vibrio was not isolated in any of the other materials, and this leads one to believe that the preventive measures taken immediately were sufficient to avoid foodstuffs being contaminated. Ann Sclavo, 1975 May-Jun, 17(3), 460 - 72 {Preliminary studies on anti-Brucella agglutinins and on immunoglobulins in subjects affected by cholera during the 1973 epidemic in the province of Bari}; Iandolo E et al.; The behaviour of brucella agglutinins in sera from 70 cholera patients has been investigated . Agglutinins for brucella, at titers of 1/40 or higher, occurred in 45 patients (64,2%); it has also been showed an antibody rise in 41 subjects with a peak titer during the second week, just like cholera agglutinins . This fact confirms the antigenic interrelationship between Vibrio cholerae and brucella group . The serum immunoglobulins, during the course of infection, have been determined . Mean levels of IgM were elevated during the second week of the disease, whereas the IgG increased during the third week, as occured in primary antibody responses . Levels of IgA in serum rose significantly during the first week of the cholera; the relations of serum IgA with S-IgA of intestinal mucosa might to explain the rapid increase of IgA in the serum. Ann Sclavo, 1975 May-Jun, 17(3), 441 - 8 {Studies on the bacteriological diagnosis of cholera and on the characteristics of isolated strains in the Apulia epidemic during the summer of 1973}; Rizzo G et al.; Descriptions are given of the characteristics of 107 strains of Vibrio cholerae of the serum type Ogawa isolated during the epidemic in 1973, which, although not exhibiting hemolysis in liquid culture, could be assigned because of their other characteristics to the El Tor biotype and to the phagic type 1 . The Authors also make a number of technical annotations on the bacteriological diagnosis of cholera. Ann Sclavo, 1975 May-Jun, 17(3), 399 - 418 {Treatment of cholera in the adult . Studies during recent epidemics in Apulia}; Pastore G; A rapid and adequate intravenous infusions of saline and alkali in correcting hypovolemic shock, alectrolyte abnormalities, acidosis and renal failure, is the choice treatment in the majority of acute cholera patients . On the basis of the knowledge of chemical composition of the gastrointestinal fluids loss and the definition of exact mechanism of metabolic features observed in acute cholera patients, it has been possible, in the last ten years, to develop and utilize in a large numbers of patients, a more exact and adequate treatment consisting of replacement of water and electrolytes depletion with saline and alkali of the same ionic composition of fluids lost through the gastrointestinal tract . The isotonic saline and isotonic sodium lactare regimen in a 2:1 ratio has been employed in the treatment of our adult acute cholera patients . Hypotensive patients received a rapid fluids infusion of 2-4 liters within the first 4-6 hours immediately after admission . Later on the quantity of intravenous fluids replacement was regulated on the basis of blood electrolytes pattern, blood pressure, diuresis and patient's hydratation . With this regimen of treatment we observed electrolytes and acid-basic abnormalities in only 25% of our patients . The peritoneal dialysis was applied in three of these patients with irreversible renal failure . Antibiotic therapy, that assume a secondary role in the treatment of cholera patients, was carried out in each case: in 19 cases were given 1 gram every 12 hours i.m . of chloramphenicol succinate; 14 cases received mg 500 of tetracycline orally every six hours; 34 cases received two tablets of trimethoprim-sulphamethoxazole every 12 hours (mg 240 of TM and g 1,6 of SMZ total dose) . Except one case of TM-SMZ group who had stool culture positive for Vibrio on 4th day of therapy, in each other patients the stool cultures were sterilized in a period of 24-72 hours of antibiotic treatment. Ann Sclavo, 1975 May-Jun, 17(3), 257 - 66 {Serological response in man to an anticholera vaccine made from killed vibrios and partially purified with aluminium hydroxide}; Soscia M et al.; The immunizing capability of a new anticholera vaccine (choleric anatoxin + vibrios Ogawa and Inaba) was tested on a group of 113 subjects . The vaccination was carried out with two inoculations of the vaccine, administered a month apart . Testings were taken before the vaccination, before the second inoculation and 10 to 14 days after the vaccination was completed . The determination of the vibriocide antibodies was made following the test-tube method described by Verwey and al., and the research of the antitoxic antibodies following Craig's method, using a live rabbit . Eighty two per cent of the subjects vaccinated had antitoxic antibodies with a varying strength of from 10 to greater than 320 A.U . In all those subjects treated who had previously been vaccinated at the time of the cholera epidemic with traditional vaccine, the geometric average of strength in vibriocide antibodies had increased from 2,810 to 14,700. Antibiotiki, 1975 May, 20(5), 456 - 8 {Action of antibiotic combinations with furacilin on the synthesis of proteins and nucleic acids in intact NAG-vibrio cells}; Belizhenko VD; Furacillin in combination with such antibiotics as tetracycline, levomycetin or neomycin inhibited the synthesis of proteins in the cells of NAG-vibrios . Combination of 8 gamma/ml of furacillin and 0.125 gamma/ml of tetracycline inhibited the protein synthesis by 58.8 per cent, 8 gamma/ml of furacillin and 0.5 gamma/ml of levomycetin inhibited the synthesis by 61 per cent, 8 gamma/ml of furacillin and 4 gamma/ml of neomycin inhibited it by 59.5 per cent . At the same time furacillin alone in concentrations of 16 and 8 gamma/ml inhibited the protein synthesis by 69.1 and 37 per cent respectively, tetracycline alone in doses of 0.25 and 0.125 gamma/ml inhibited it by 51.3 and 34.7 per cent respectively, levomycetin alone in doses of 1 and 0.5 gamma/ml inhibited it by 54.4 and 33.2 per cent, enomycin in doses of 8 and 4 gamma/ml inhibited it by 54.4 and 22.6 per cent respectively . Therefore, when the above antibiotics were used in combination with furacillin the inhibitory effect of the drugs on the protein synthesis was summarized . When furacillin was combined with tetracycline or levomycetin in the above concentrations, the inhibitory effect on RNA synthesis (42 or 32 per cent respectively) was lower than that of furacillin alone (50.5 per cent) . When furacillin was used in combination with neomycin, the inhibition of RNA synthesis increased up to 69 per cent . The increase in the inhibitory effect was also noted with respect to the synthesis of DNA . Combination of furacillin with tetracycline, levomycetin or neomhcin decreased the synthesis of DNA by 79.7, 85 or 85.8 per cent respectively as compared to the inhibitory effect of 8 gamma/ml of furacillin equal to 61 per cent. Zh Mikrobiol Epidemiol Immunobiol, 1975 May, (5), 94 - 9 {Role of neuraminidase from cholera vibrions in the pathogenesis of experimental cholera}; Solov'ev VD et al.; A study was made of a possible effect of neuraminidase of cholera vibrios on cholera pathogenesis . It was shown that in intraintestinal injection of cholera vibrios of the El Tor biotype to nursling rabbits neuraminidase could be revealed in their intestine 5 to 8 hours after the infection . Addition of neuraminidase to the weakly cholerogenic strain cholera vibrios intensified its cholerogenic action in infection of the animals . The antineuraminidase serum administered to the infected rabbits prevented clinical manifestations of experimental cholera, although it failed to always eliminate the cholerogenic syndrome (revealed during autopsy) . At the same time neuraminidase did not influence the capacity of cholerogen to produce the cholerogenic syndrome . The authors consider that the action of the enzyme should occur at the early stages of the pathogenic process, and could be associated with creation of conditions for the attachement of cholera causative agent to the intestinal wall or for the action of their exotoxin. Appl Microbiol, 1975 May, 29(5), 635 - 40 Isolation and characterization of Vibrio parahaemolyticus from Cape Cod soft-shell clams (Mya arenaria); Earle PM et al.; Vibrio parahaemolyticus was isolated from soft-shell clams (Mya arenaria) taken from 10 different clamming areas on Cape Cod, Mass., during July and August 1972 . Direct plating on thiosulfate-citrate-bile salts-sucrose agar was found to be superior to either direct plating on Vanderzant modified salt starch agar or enrichment with Trypticase soy broth containing 7% salt for isolation from clam samples . Morphological and biochemical characteristics of 33 isolates from 30 samples generally conform to those described for this organism in the literature, except for the production of acid from sucrose, lactose, and sorbitol . Six of the isolates were hemolytic on human blood agar plates, whereas all showed a negative Kanagawa phenomenon . Twenty of the 33 isolates reacted with pooled antisera to the K antigen; 15 of these reacted with 9 different specific K antisera, leaving 5 untypable . Ten of these 15 reacted with 4 different O antisera. J Bacteriol, 1975 May