Covalently protein-bound bilirubin conjugates in cholestatic disease of dogs

We investigated the occurrence of covalently protein-bound bilirubins in the plasma of dogs with hyperbilirubinemia attributable to hepatobiliary diseases or Coombs test-positive hemolysis. The bilirubins in plasma were measured with the conventional Van den Bergh reaction, by treatment with diazotized p-iodoaniline, and by high-performance liquid chromatography of bilirubin and its methylesters after alkaline methanolysis. All but one dog had covalently protein-bound bilirubin conjugates. The concentration and the fraction of total bilirubins varied in all diseases investigated, but they tended to be low in primary hemolysis. The "biliprotein" complex accounted for 2 to 94% of total plasma bilirubins. Because biliprotein usually is not cleared by the liver, but has a half-life comparable with that of albumin, it prevents the evaluation of the actual state of the underlying disease. Measurement of the total bilirubin concentration exclusively with the Van den Bergh reaction, therefore, is clinically useless. Other methods should be introduced for routine bilirubin assays, permitting the measurement of noncovalently bound pigment as a meaningful estimate of the course of the disease.     

A blind test of the serological response of dogs to infection with Taenia ovis

Fifty six dogs of mixed age and sex were acquired from farms in the Otago/Southland region, and maintained at the Hydatid Research Unit, Taieri, where 43 were each fed two Tueniu ovis cysts. All were bled fortnightly for six or 12 weeks. Coded sera were sent to Wallaceville Animal Research Centre for testing using ELISA, with antigen from T. ovis scoleces. Dog treatments were identified after all tests were complete. A discriminant level was derived from the mean absorbance value plus three standard deviations of 56 sera taken at time zero and 78 sera from serially bled uninfected dogs. None of these 134 sera registered as a false positive using this discriminant level. The data showed no significant deviation from normality, and the expected frequency of the occurrence of false positives is therefore less than 0.14%. Four weeks after infection 63% of dogs proved to be infected were serologically positive, rising to 78% after 6 weeks. When worms were removed by anthelmintic treatment, ELISA absorbance levels decreased. Four weeks after removal 70% of previously infected dogs remained positive, decreasing to 30% after 6 weeks. Six weeks after infection the sensitivity of the test was 78%, and the specificity 63%. However, if dogs with positive ELISA absorbance levels, but which did not purge worms, were regarded as having had worms, the respective figures would be 82% and 100%. The latter figures are similar to our previously published laboratory results. The test is of comparable efficiency to arecoline purgation for surveillance, and has the additional advantage of detecting infection in the majority of those dogs that have been infected for three weeks or more but fail to pass worms on purgation, and a substantial proportion of those infected dogs that were treated by their owners prior to presenting them for purgation in order to avoid detection of infection.     

The detection of Mycobacterium paratuberculosis in bovine faeces by isolation and the comparison of isolation with the examination of stained smears by light microscopy

The detection of Mycobacterium paratuberculosis organisms in bovine faeces by isolation was compared with that by the microscopical examination of Ziehl-Neelsen stained faecal smears for the presence of clumps of acid-fast M. paratuberculosis organisms. Faeces were obtained from cattle naturally or experimentally infected with M. paratuberculosis as well as from uninfected cattle. Microscopical examination was an unreliable method for the detection of M. paratuberculosis organisms, since the organisms were only detected in 99 (=55.9%) of 177 culturally positive faecal samples. 1111 addition, clumps of acid-fast organisms indistinguishable from M. paratuberculosis were also observed iin three of 18 samples from cattle free from Johne's disease and in 18 of 37 culturally negative samples from paratuberculous cattle. When M. paratuberculosis organisms were added to faeces from an uninfected cow, results showed that isolation attempts should be positive when 15 or more M. paratuberculosis organisms per gram of faeces are present.