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Microtitration agglutination for detection of Treponema hyodysenteriae antibody.

A microtitration agglutination test for the detection of Treponema hyodysenteriae antibody in swine and rabbit sera is described. The following methods provided the best test results: antigen produced from the spirochete after a culturing period of 36 to 44 h at 38 degrees C, washed antigen inactivated with 0.01% Merthiolate at 4 degrees C for 24 to 36 h, sera heated at 56 degrees C for 30 min, a diluent of phosphate-buffered saline (0.01 M, pH 7.2), and test results read macroscopically after 18 to 24 h of incubation at 38 degrees C. The test enabled detection of antibody against pathogenic T. hyodysenteriae with a high level of consistency and sensitivity. Sera against nonpathogenic T. hyodysenteriae produced low agglutinating titers (less than or equal to 1:8) when reacted against antigen from pathogenic isolates. Inactivated antigen remained stable for 7 to 10 days. Specificity of the reaction in the agglutination test was shown by absorption studies.

Agglutination Tests↗

Factors affecting the survival of Treponema hyodysenteriae in dysenteric pig faeces.

Treponema hyodysenteriae was found to survive for periods of up to 48 days in dysenteric pig faeces stored at temperatures between 0 degree C and 10 degree C inclusive. Survival was reduced to seven days at 25 degree C and did not exceed 24 hours at 37 degree C. Dilution 1:10 with tapwater appeared to enhance survival to a maximum of 61 days at 5 degree C but further dilution reduced it. Drying and exposure to disinfectants rapidly eliminated T hyodysenteriae from dysenteric faeces. Phenolic and sodium hypochlorite disinfectants were most effective. The use of these findings in the formulation of control programmes for swine dysentery is discussed.

Animals↗

Sensitivity in vitro to dimetridazole of treponemes associated with swine dysentery.

The minimum inhibitory concentration (MIC) of dimetridazole (DMZ) against Treponema hyodysenteriae (55 isolates) obtained over the period 1974-77 from individual pigs with swine dysentery from 41 herds where DMZ had been in use was determined. The MIC was less than or equal to 5.0 microgram per ml for 54 of the isolates and differences in the distribution of MICs between the annual sampling periods were not significant (P less than 0.05). There was no decrease in sensitivity of T hyodysenteriae to DMZ during the survey.

Animals↗

Comparison of selective culture and serologic agglutination of Treponema hyodysenteriae for diagnosis of swine dysentery.

Samples of faeces and serum were collected from pigs of various ages on 21 farms. Faecal samples were cultured on trypticase soy agar containing 5% citrated bovine blood and 400 microgram per ml spectinomycin, incubated at 42 degrees C in Gaspak jars under an atmosphere of 80% hydrogen: 20% carbon dioxide. Antibody titres to Treponema hyodysenteriae were determined by a microtitration agglutination method using merthiolate-inactivated whole cell antigen prepared from a beta- haemolytic isolate. Results indicated that mean titres in pigs from which beta- haemolytic T hyodysenteriae was isolated were significantly higher than in pigs which yielded isolates of weak beta- haemolytic T innocens or in culturally negative pigs (P less than 0.0225). Mean titres of herds where beta- haemolytic T hyodysenteriae was isolated were significantly higher (P less than 0.005) than the mean titres of either of the other two groups. However, mean titres of herds where no isolates were obtained were not significantly different from mean titres of herds where weak beta- haemolytic T innocens was isolated.

Agglutination Tests↗

Production of diarrhoea and dysentery in pigs by feeding pure cultures of a spirochaete differing from Treponema hyodysenteriae.

A weakly beta-haemolytic spirochaete, isolate P43/6/78, was isolated from a pig with diarrhoea and found not to fluoresce with a specific fluorescent antiserum to Treponema hyodysenteriae. Pure cultures of this spirochaete were used to inoculate experimental pigs. Diarrhoea, containing clear mucus, and, in one case, blood occurred in four of the eight animals inoculated. Colitis was present in six of the eight inoculated pigs at necropsy. Excess clear mucus and punctate haemorrhages were seen on the colonic mucosa and spirochaetes resembling isolate P43/6/78 were reisolated from the affected mucosa. The feed conversion efficiency and growth rate of affected pigs was reduced when compared with controls. Isolate P43/6/78 differed from T hyodysenteriae in its cultural, ultrastructural, biochemical and antigenic characters. On these grounds, and because of the clinical and pathological syndrome produced, it was considered to belong to a species other than T hyodysenteriae.

Animals↗

Diagnosis of swine dysentery: spirochaetes which may be confused with Treponema hyodysenteriae.

Spirochaetes, designated PWS, microscopically resembling Treponema hyodysenteriae, were isolated from the colon contents of pigs with post-weaning scours from three herds, which on clinical and epidemiological grounds were thought to be free from swine dysentery. One of the isolates was fed to pigs experimentally but no evidence of disease was noted. Their cultural characteristics differed from those of T hyodysenteriae but they were similar to those of a non-pathogenic spirachaete (4/71) which had been isolated previously from the laboratory's dysentery-free herd. Smears prepared from cultures of the PWS spirochaete, 4/71 and two virulent and one avirulent strain of T hyodysenteriae were all positive to a fluorescent antibody test which was in use as an aid to the diagnosis of swine dysentery in the United Kingdom.

Animals↗

Studies on the pathogenesis of swine dysentery. I. Characterization of the lesions in colons and colonic segments inoculated with pure cultures or colonic content containing Treponema hyodysenteriae.

Swine dysentery was induced in pigs and in ligated colonic segments by inoculation of pure cultures of, or colonic contents containing, Treponema hyodysenteriae. The mildest changes, best seen in ligated segments 48 or 72 hours after inoculation, were congestion and leucocytic margination in mucosal capillaries and depletion of mucigen from goblet cells lining the base of the crypts of Lieberkühn. Superficial mucosal necrosis and crypt cell hyperplasia were later changes. Perfusion studies with India ink did not demonstrate occlusive mucosal ischemia in acute swine dysentery. Mucosa with lesions of swine dysentery contained at least 10(5) colony forming units of T. hyodysenteriae per gram. Mucosa without lesions had 10(5) or fewer T. hyodysenteriae per gram. Segments with acute swine dysentery were distended with clear mucoid fluid with electrolyte composition indicative of net colonic secretion. No increase in the concentration of volatile fatty acids was detected in content from intact colons or colonic segments with lesions of acute swine dysentery.

Animals↗