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Biomedical subjects

C L Gyles

Publications and source records attributed to C L Gyles.

At least 91 records · Page 5Linked to original sources

The effect of adsorbant and anti-inflammatory drugs on secretion in ligated segments of pig intestine infected with Escherichia coli.

Four adsorbant drug preparations, Kaopectate, colloidal Attapulgite, noncolloidal Attapulgite and Pepto-bismol were investigated for their effects on fluid accumulation in ligated segments of pig intestine inoculated with enteropathogenic Escherichia coli. Two anti-inflammatory drugs. aspirin and methylprednisolone, and two antibiotics, lincomycin and polymyxin B, were also tested. All the drugs except the two anti-inflammatory products were given by injection into the lumen of the intestine. Aspirin was given orally and methylprednisolone was given intramuscularly. The antibiotics were tested at levels at which they had no significant antibacterial effect in in vitro tests. The adsorbant drugs colloidal Attapulgite and Pepto-bismol were shown to be effective in reducing fluid accumulation in ligated segments of pig intestine infected with enteropathogenic E. coli. In the case of Peptobismol this effect was associated with an antibacterial effect as well as an antitoxic effect, probably due to its adsorbant properties. It is possible that an aspirin-like effect in the gut due to the active ingredient bismuth subsalicylate may have contributed to the effectiveness of Pepto-bismol. Colloidal Attapulgite was demonstrated to have an antitoxic effect but did not have an antibacterial effect. In high doses, the anti-inflammatory drugs acetylsalicylic acid and methylprednisolone were marginally effective in reduction of fluid accumulation in the same test system. Lincomycin was shown to reduce intestinal fluid secretion, whereas polymyxin B had no effect.

Animals↗

Naturally occurring plasmid carrying genes for enterotoxin production and drug resistance.

Escherichia coli strain 86, isolated from a piglet with diarrhea, carries plasmid-linked genes for resistance to tetracycline, streptomycin, and sulfonamides and for production of heat-labile and heat-stable enterotoxin. Results of (i) genetic experiments involving conjugal transfer and phage P1-mediated transduction and (ii) physical experiments involving electron microscopic examination of plasmid DNA and heteroduplex analysis show that a single conjugative plasmid carries the genes for drug resistance and production of enterotoxin.

Conjugation, Genetic↗

The prevalence of enterotoxigenic Escherichia coli in the feces of calves with diarrhea.

A study was undertaken to determine the prevalence of enterotoxigenicity among Escherichia coli isolated from calves with diarrhea and from a control group of normal calves. The test organisms consisted of 200 E. coli recovered from scouring calves less than two weeks of age and 100 E. coli from normal calves. The enterotoxigenicity of the cultures was evaluated by three methods, namely, injection of ligated segments of piglet intestine, injection of ligated segments of calf intestine and oral inoculation of suckling mice. Live cutures of all the test organisms were used for the ligated intestine studies whereas sterile broth culture supernatants were used in the suckling mouse tests. Of the isolates from scouring calves, 36% were enterotoxigenic in the piglet intestine and 28% in the calf intestine. Amongst the isolates from normal calves, none was enterotoxigenic in the piglet intestine and one was enterotoxigenic in the calf test system. The ligated piglet intestine was considered unsuitable for determining the enterotoxigenicity of bovine E. coli, whereas the ligated calf intestine test was satisfactory and correlated completely with the suckling mouse test. The enterotoxigenic E. coli of bovine origin produced an enterotoxin that resembled the heat stable enterotoxin of typical porcine enteropathogenic E. coli.

Animals↗

Characterization of enterotoxigenic bovine Escherichia coli.

Among 300 isolates of bovine Escherichia coli, 56 which had been found enterotoxigenic in calf gut loops were characterized on the basis of O and K antigens, colonial morphology and resistance to seven antimicrobial drugs. The 56 isolates enterotoxigenic in the calf were compared with the nonenterotoxigenic ones. Of the 56 enterotoxigenic E. coli the majority possessed the A type of K antigen and had OK groups, O9:K(PS274) or O101:K(RVC118). Fourteen of these isolates had the K99 antigen. None of 27 isolates found enterotoxigenic in the piglet but not in the calf possessed the K99 antigen or belonged to OK groups O9:K(PS274) or O101:K(RVC118). Comparison of the patterns of resistance to seven antimicrobial drugs showed that all enterotoxigenic and nonenterotoxigenic isolates were susceptible to nitrofurantoin and sulphachlorphyridiazine and that there was no significant difference in the patterns between the two groups. The majority of enterotoxigenic isolates were mucoid, whereas most of the nonenterotoxigenic isolates were nonmucoid.

Animals↗

The effect of antisera on porcine enteropathogenic Escherichia coli in ligated segments of pig intestine.

Nineteen antisera produced in pigs against 14 enteropathogenic and five nonenterotoxigenic porcine strains of Escherichia coli were tested for their ability to inhibit gut loop fluid accumulation induced by homologous and heterologous organisms. In addition, four antisera produced in pigs by an intensive series of intravenous inoculations and three by a less intensive series of intramuscular injections of a polyvalent E. coli vaccine were evaluated. Antisera were also produced in rabbits against eight strains of porcine enteropathogens and tested in pig gut loops. Fluid inhibiting activity was detected in prevaccinal sera of pigs but not of rabbits. This activity was significantly increased following immunization. When single strains of E. coli were used for immunization the activity of the antisera against heterologous organisms varied considerably from one test strain to another and was usually much less than that against the homologous organism. The activity against heterologous organisms could not be associated with relatedness of the O, K and H antigens of the vaccine and the test strains. Antisera produced against a vaccine made by combining three strains were shown to exert inhibitory effects on heterologous organisms similar to those against homologous organisms. Considerably less activity against homologous and heterologous organisms was present in antisera produced by the series of intramuscular compared with the series of intravenous injections.

Animals↗

Antibacterial activity of antisera against homologous and heterologous Escherichia coli of porcine origin.

Fourteen enteropathogenic and five nonenterotoxigenic Escherichia coli strains isolated from pigs were used for producing antisera in rabbits and pigs. These antisera were used in an vitro test system for antibacterial activity against homologous and heterologous porcine E. coli strains. Antibacterial titres were determined against the homologous strains and the percent reduction in CFU/ml caused by a 1/200 dilution of the sera against heterologous strains was determined. The results indicated that following immunization the antibacterial activity of serum against homologous and heterologous strains was significantly increased. This activity did not appear to be influenced by O and K antigen relationships among the organisms or by enterotoxigenicity of the vaccine strains. When antiserum produced against a combination of three enteropathogenic E. coli was tested against 20 strains a wider spectrum of heterologous antibacterial activity was obtained than with antiserum produced against any individual strain. The results indicate the existence in E. coli strains of porcine origin of common antigenic determinants not related to the serological formula and that a selected combination of strains can be expected to induce antibacterial acitivity against a wide variety of serological types of porcine enteropathogenic E. coli.

Absorption↗

Immunological study of the heat-labile enterotoxins of Escherichia coli and Vibrio cholerae.

Immunodiffusion experiments were conducted to associate a precipitin line with Escherichia coli heat-labile enterotoxin (LT). Wild strains of porcine and of human enteropathogenic E. coli as well as laboratory-derived enterotoxigenic variants of E. coli K-12 were used for LT antigen preparations. These were produced mainly by ultrafiltration and ammonium sulfate precipitation of broth culture supernatants. When antisera with anti-LT activity were reacted with antigen preparations from Ent(+) and Ent(-) variants of E. coli K-12, a line "a" was given by Ent(+) but not by Ent(-) preparations. Line "a" was removed by absorption of anti-LT serum with antigen preparation from an Ent(+)E. coli K-12, but was unaffected when the antigen preparation used to absorb the serum was from an Ent(-)E. coli K-12. A line identical to "a" was given by antigen preparations from wild strains of porcine enteropathogenic E. coli reacted with homologous or heterologous anti-LT sera. One human strain of enteropathogenic E. coli was shown to possess an antigen identical to that which gave rise to line "a." To demonstrate this line it was necessary to use high concentrations of gammaglobulin and high concentrations of the crude antigen preparations. LT preparations reacted with anticholera toxin to give a line "c," which showed a reaction of partial identity with line "b" produced by reaction of pure choleragenoid and anticholera toxin. Lines "a" and "c" gave reactions of identity.

Animals↗

Isolation of Escherichia coli O157 from pigs with colibacillosis in Canada and the United States.

Cultures of Escherichia coli isolated from colibacillosis in pigs in Canada and the U.S.A. and previously identified as unclassified E. coli OX36 were found to belong to a new E. coli O group 157. Strains of E. coli V145, GIRI, C308 and V17 which had a weak relationship to E. coli O116 also were found to belong to O157. The increased incidence of E. coli O157 in colibacillosis of swine stresses the inclusion of this O group among those enteropathogenic for swine.

Agglutination Tests↗

The significance of proliferation and enterotoxin production by Escherichia coli in the intestine of gnotobiotic pigs.

The significance of enterotoxin production and proliferative ability of Escherichia coli in the intestinal tract as related to porcine enteric colibacillosis was studied in 68 gnotobiotic pigs. The animals were monocontaminated at seven to ten days of age with eight selected strains of E. coli. The strains were two naturally occurring porcine enteropathogens - P155 (0149:K91;K88a,c:H10) and P307 (08:K87;K88a,b:H19), two nonenteropathogenic strains - P104 (0139:K82:H1) and F11 (018-ab:K?:H14), and four enterotoxigenic derivatives of the above strains - P104(P155), P104(P307), F11(P155) and F11(P307). The response of the animals was evaluated on the basis of clinical observations and necropsy lesions 22 hours after exposure to the organisms. E.coli counts were determined at seven different levels of the intestinal tract. Cell free extracts of the intestinal contents were examined for enterotoxic activity by the ligated pig intestine loop test. All of the strains possessing the enterotoxin plasmid produced enterotoxin in the pig's intestine and were capable of causing diarrhea. The nonenteropathogenic E. coli failed to do so. The strains possessing the P155 enterotoxin plasmid were more virulent than the corresponding derivatives with the P307 enterotoxin plasmid. Strains P155, P307 and P104(P155) proliferated in the upper small intestine at a greater rate and were more virulent than the other strains. The numbers attained in the upper small intestine by the other enterotoxigenic derivatives were comparable to those of their nonenteropathogenic parent strains. It was considered that enterotoxin produced by E. coli was the essential factor for causing a diarrheic response in gnotobiotic pigs. The virulence of each of the tested strains appeared to be governed by the degree of enterotoxicity associated with a particular enterotoxin plasmid, the numbers attained by these organisms in the upper small intestine, (but not in the lower small intestine or in the colon), and by other undetermined factors.

Animals↗

A comparative study of the rabbit and pig gut loop systems for the assay of Escherichia coli enterotoxin.

A comparison was made between segments of pig and rabbit small intestine in their response to heat-labile (LT) and heat-stable (ST) preparations from porcine enteropathogenic Escherichia coli. Either whole cell lysates or dialysed broth culture supernatants were used as sources of LT and soft agar culture fluids as a source of ST. Whole cell lysates of all thirteen LT-producing E. coli strains tested regularly elicited fluid accumulation in rabbit gut loops. Whole cell lysates of certain E. coli strains considered to be nonenteropathogenic in pigs could also elicit a positive response in rabbit gut loops. When graded doses of LT were tested in pig and rabbit gut loops, the rabbit was more sensitive and is therefore considered preferable to the pig for quantitation of LT. In the rabbit, upper (jejunal) and lower (ileal) small intestine were compared for their response to LT and it was found that ileal loops were twice as sensitive but more prone to false positive reactions. When soft agar culture fluids of several enteropathogenic E. coli strains were tested in the rabbit, the response was inconsistent, and it was concluded that the rabbit is unsuitable for the assay of the heat-stable enterotoxin.

Animals↗

A study of Escherichia coli strains isolated from pigs with gastro-intestinal disease.

When 200 Escherichia coli isolated from pigs with gastro-intestinal disease were examined, 82.0% of them were found to be enteropathogenic by the ligated intestine test in pigs. Among the 18.0% which were nonenteropathogenic, 12.0% were obtained from pigs considered likely to be suffering from colibacillosis and 6.0% were from pigs for which there was a satisfactory diagnosis. Eighty-seven percent of the enteropathogens belonged to the serogroups O8:K87;88ac, O116:K"V17";88ac, O147:K89;88ac, O138:K81 and O45:K"E65";88ac. One O4 isolate and two O98 isolates were enteropathogenic, although members of these O groups have not been recognized enteropathogenic types. The majority (153) of the enteropathogens were of the types which are well characterized and which induce fluid accumulation in ligated segments of the intestine of young and old pigs and were referred to as class 1 enteropathogens. A small number (11)were able to cause fluid accumulation in ligated segments of young, but not of older pigs and were referred to as class 2 enteropathogens. Enterotoxin production was demonstrated for strains of all the serogroups of class 1 enteropathogens detected, and transfer of the enterotoxin plasmid was carried out for strains of O groups 4, 45, 98, 116 and 147.

Age Factors↗