Variability of serum bile acid concentrations over time in dairy cattle, and effect of feed deprivation on the variability.

Twelve nonlactating dairy cows, free of signs of liver disease and with normal serum activities of liver-derived enzymes and normal liver biopsy tissue, were examined over a 72-hour period for serum total bile acid concentrations. The cattle were fed hay twice daily, and blood samples were obtained every hour for 24 hours, every other hour for 24 hours, then every hour for 24 hours. After 3 weeks, the study was repeated on 6 of the cattle, thus providing data for eighteen 72-hour periods. Serum bile acid concentration varied greatly over the 72 hours, with the range being from one third to 3 times the median. There were variations by as much as 60 mumol/L from 1 hour to the next. After another 3 weeks, 8 of the cattle were deprived of hay for 48 hours and then fed hay morning and afternoon of the third (last) day of the study. There was no significant reduction in bile acid concentration after withholding the hay, but the variability was reduced (P = 0.02) during the last 20 hours of the hay-deprivation period. In 3 ancillary studies, serum bile acid concentrations were examined over a 48-hour period in 2 cows in early lactation, 3 cows in midlactation, and two 6-month-old heifers. The cows were fed hay and grain twice daily, and the heifers were fed only hay twice daily. In comparison with values for the 12 nonlactating cows fed hay twice daily, mean serum bile acid concentration in the recently freshened cows was significantly (P < 0.002) higher (62.9 vs 22.0 mumol/L).(ABSTRACT TRUNCATED AT 250 WORDS)     

Multiresidue assay for benzimidazole anthelmintics by liquid chromatography and confirmation by gas chromatography/selected-ion monitoring electron impact mass spectrometry

A multiresidue method utilizing all-disposable labware has been developed for 8 benzimidazole anthelmintics from ovine, bovine, and swine muscle and liver tissues. After an initial extraction with ethyl acetate and subsequent evaporation, a 3-component extraction using hexane, ethanol, and 0.2N HCl was used for final cleanup. Clean extracts were produced for separation and determination by reverse-phase liquid chromatography at 298 nm, using methanol and aqueous buffer as mobile phase. A synthesized internal standard, 2-(n-butylmercapto)benzimidazole, was used for quantitation of all drugs. Results are included along with statistical information verifying the performance of the method. Spiked control tissues and incurred drug tissues were used for an intralaboratory study with a concentration range of 50-1470 ppb. A series of standard curves at 0, 50, 100, and 200 ppb were analyzed. Overall recovery at the 100 ppb level averaged 92% (CV 8%) in liver tissues, across all 3 species and 88% (CV 5%) in muscle tissues across all 3 species. Results were confirmed by gas chromatography/mass spectrometry with acid hydrolysis of the remaining extract in 2N HCl followed by re-extraction of the amine and derivatization to the tert-butyldimethylsilyl derivative. The anthelmintics were identified by gas chromatography/selected ion monitoring electron-impact mass spectrometry. Ion ratio measurements were taken and compared to standard material. CVs averaged 10% or less for all drugs tested.     

Physiological effects of arginine vasotocin and mesotocin in cockerels

1. The effects of continuous infusion of 0.1, 1.0 and 10.0 mU/min/kg body weight of arginine vasotocin (AVT) or mesotocin (MT) on cardiovascular and thermoregulatory responses, on plasma osmolality and ionic composition and on plasma concentrations of AVT, MT, prolactin and aldosterone, were investigated in conscious White Leghorn cockerels.

2. Neither of the peptides, at any dose, affected cardiovascular functions, plasma ions and osmolality. Infusion of MT at the rate of 10 mU/min/kg body weight increased respiratory rate. Both peptides at doses of 1 and 10 mU/min/kg reduced the temperatures of the comb and shank but had no effect on the skin and cloaca.

3. Doses of 0.1 and 1.0 mU MT/mu/kg reduced plasma aldosterone and at 10 mU/min/kg increased plasma AVT. At any given dose MT had no effect on plasma prolactin. AVT at 0.1 and 1.0 mU/min/kg of AVT reduced plasma MT. AVT at 1.0 mU/min/kg increased plasma prolactin and at 10 mU/min/kg reduced plasma aldosterone.

4. During saline infusion, plasma MT was positively correlated with plasma AVT and negatively correlated with respiratory rate and cloacal temperature. Plasma AVT showed a positive correlation with plasma MT and aldosterone and a negative correlation with respiratory rate and skin temperature.

5. During saline infusion, there was no significant correlation between cardiovascular functions, or plasma osmolality and ionic composition and plasma MT or AVT.

6. The present study suggests that interrelationships between circulating concentrations of AVT and MT do exist and that AVT affects     

A framework for evaluation of on-farm mastitis diagnostics in Australia

Numerous culture-based diagnostics are available on the Australian and international markets for on-farm detection of bacterial pathogens in milk. Use of such diagnostics may provide an opportunity to improve the prudent use of antimicrobials in udder health management. Farms are low-resource settings in terms of diagnostic microbiology capacity. The World Health Organisation has identified criteria for the evaluation of diagnostic tests in low resource settings based on Accuracy, Sensitivity, Specificity, User-friendliness, being Rapid or Robust, Equipment-free and being Deliverable (ASSURED). Here, we review how those criteria can be interpreted in the context of microbiological diagnosis of mastitis pathogens, and how on-farm diagnostics that are currently available in Australia perform relative to ASSURED criteria. This evaluation identifies multiple trade-offs, both with regard to scientific criteria and with regards to convenience criteria. More importantly, the purpose of testing may differ between farms, and test performance should be evaluated relative to its intended use. The ability of on-farm mastitis diagnostics to inform mastitis treatment decision-making in a timely and cost-effective manner depends not just on test characteristics but also on farm-specific pathogen prevalence, and on the farm enterprise's priorities and the farm manager's potential courses of action. With most assay evaluations to date conducted in professional laboratories, there is a surprising dearth of information on how well any of the diagnostic tests perform on-farm and, indeed, of the on-farm decision-making processes that they aim to inform.     

Phage sensitivity in relation to pathogenicity and virulence of the cotton bacterial blight pathogen of Sudan

At least two pathotypes of Xanthomonas campestris pv. malvacearum are now known to exist in Sudan. The pre-Barakat (race 1) and post-Barakat (race 2) pathogens have been shown to exhibit different host specificity. The former is pathogenic and highly aggressive on only cultivars with no resistance genes or with the B₂ and/or B₃ resistance factors, while the latter can infect the B₆ cultivars also. Race 2 in Sudan, which was previously reported to infect all the standard differentials, produced milder angular leaf spot symptoms and occasionally restricted vein infection. Moreover, it exhibited reduced growth in planta compared with race 1.
Bacteriophage studies revealed that the two races are quite distinct in their phage sensitivity. Race 1 can be lysed by only three, or rarely four, of the six phages used for typing, while race 2 is sensitive to all of them. The present study suggests that phage 7 may be the type-determining phage for race 2. Race 2 strain mutants resistant to phage 3 or 4 were found to be sensitive to phage 7 and pathogenic to both Acala and Barakat, although showing marked attenuation of virulence. However, mutants resistant to phage 2 or 7 were insensitive to all the phages and although they retained their pathogenicity to Acala, they either lost the ability to infect Barakat or produced a hypersensitive reaction. The resistance of all mutants was found to be due to failure to adsorb the homologous phage, indicating a change in the cell wall. The association of this with the attenuation of virulence suggests that bacterial wall components may function as virulence determinants in Xanthomonas campestris pv. malvacearum.     

Isolation and characterisation of cervine herpesvirus-1 from red deer semen

AIM: This communication describes the isolation of herpesvirus during routine export examination of semen collected from red deer stags in New Zealand. METHODS: Virus isolation was carried out using bovine embryonic lung (BEL) cells and viruses were characterised by direct immunofluorescense, restriction-fragment-length polymorphism analysis (RFLP), polymerase chain reaction (PCR) analysis and nucleotide sequencing. RESULTS: Herpesvirus was isolated from red deer semen on 2 different occasions from different animals. In both cases the virus was identified as cervine herpesvirus-1 (CvHV-1), based on RFLP, PCR and sequence analysis. Nucleotide sequence analysis of the glycoprotein-D gene showed 99.7% homology to the Banffshire strain of CvHV-1 and 89.5%, 89.2%, 85.3% and 79.6% homology to bovine herpesvirus 1.2 (BoHV-1.2), bovine herpesvirus 1.1 (BoHV-1.1), cervine herpesvirus-2 (CvHV-2) and caprine herpesvirus-1 (CpHV-1), respectively. CONCLUSION: This is the first time that CvHV-1 has been isolated in New Zealand. Its inclusion in serological surveys will allow the prevalence of CvHV-1 in the red deer population to be assessed in this country. The clinical significance of CvHV1 infection in New Zealand red deer herds has yet to be determined.