Haematological and biochemical reference values for the koala (Phascolarctos cinereus)

Haematological and biochemical reference values were established from 45 clinically healthy koalas. Statistical analysis revealed no significant differences for sex and season of sampling. Immature koalas had significantly higher alkaline phosphatase and inorganic phosphate values, and significantly lower total protein concentrations due to low globulins values. Enzyme reference values tended to be wide and could limit their usefulness in detecting disease. In the reference values for leukocytes, neutrophils and lymphocytes, the inclusion of low values which were not actually seen may interfere with the detection of reduced levels due to disease.     

Genetic determination of individual birth weight and its association with sow productivity traits using Bayesian analyses

Genetic association between individual birth weight (IBW) and litter birth weight (LBW) was analyzed on records of 14,950 individual pigs born alive between 1988 and 1994 at the pig breeding farm of the University of Kiel. Dams were from three purebred lines (German Landrace, German Edelschwein, and Large White) and their crosses. Phenotypically, preweaning mortality of pigs decreased substantially from 40% for pigs with < or = 1 kg weight to less than 7% for pigs with > 1.6 kg. For these low to high birth weight categories, preweaning growth (d 21 of age) and early postweaning growth (weaning to 25 kg) increased by more than 28 and 8% per day, respectively. Bayesian analysis was performed based on direct-maternal effects models for IBW and multiple-trait direct effects models for number of pigs born in total (NOBT) and alive (NOBA) and LBW. Bayesian posterior means for direct and maternal heritability and litter proportion of variance in IBW were .09, .26, and .18, respectively. After adjustment for NOBT, these changed to .08, .22, and .09, respectively. Adjustment for NOBT reduced the direct and maternal genetic correlation from -.41 to -.22. For these direct-maternal correlations, the 95% highest posterior density intervals were -.75 to -.07, and -.58 to .17 before and after adjustment for NOBT. Adjustment for NOBT was found to be necessary to obtain unbiased estimates of genetic effects for IBW. The relationship between IBW and NOBT, and thus the adjustment, was linear with a decrease in IBW of 44 g per additionally born pig. For litter traits, direct heritabilities were .10, .08, and .08 for NOBT, NOBA, and LBW, respectively. After adjustment of LBW for NOBA the heritability changed to .43. Expected variance components for LBW derived from estimates of IBW revealed that genetic and environmental covariances between full-sibs and variation in litter size resulted in the large deviation of maternal heritability for IBW and its equivalent estimate for LBW. These covariances among full-sibs could not be estimated if only LBW were recorded. Therefore, selection for increased IBW is recommended, with the opportunity to improve both direct and maternal genetic effects of birth weight of pigs and, thus, their vitality and pre- and postnatal growth.     

Comparison of a competitive inhibition ELISA and the card agglutination test for detection of antibodies to Anaplasma marginale and Anaplasma centrale in cattle

OBJECTIVE: To compare a recently developed recombinant MSP-5 competitive inhibition ELISA with a card agglutination test for detection of antibodies to Anaplasma marginale and Anaplasma centrale in Australian cattle. MATERIALS AND METHODS: The ELISA was compared with the card agglutination test using 208 sera from cattle in Anaplasma-free herds, 86 sera from cattle experimentally infected with A marginale or A centrale and 757 sera from cattle in areas endemic for A marginale. RESULTS: The specificity of the ELISA, based on testing 208 sera from cattle in Anaplasma-free areas, was 99.5%, and the sensitivities for detection of antibodies to A marginale and A centrale in sera from the experimentally infected cattle were 98.0% and 100%, respectively. For the same sets of sera, the specificity of the card agglutination test was 98.6% and the sensitivities for detection of antibodies to A marginale and A centrale were 98.0% and 100%, respectively. For the 757 sera collected from cattle in areas endemic for A marginale, the agreement between the ELISA and the card agglutination test depended on the positive threshold selected for the ELISA. The maximum achievable agreement was 91.5% (kappa = 0.73; 95% confidence interval 0.66, 0.79). CONCLUSION: We conclude that the competitive inhibition ELISA is a useful alternative to the card agglutination test for detection of A marginale or A centrale infection in cattle. The assay should be particularly useful for epidemiological applications such as prevalence studies and control programs.     

Comparison of repeatability and multiple trait threshold models for litter size in sheep using observed and simulated data in Bayesian analyses

Bayesian analyses were used to estimate genetic parameters on 5580 records of litter size in the first four parities from 1758 Mule ewes. To examine the appropriateness of fitting repeatability (RM) or multiple trait threshold models (MTM) to litter size of different parities, both models were used to estimate genetic parameters on the observed data and were thereafter compared in a simulation study. Posterior means of the heritabilities of litter size in different parities using a MTM ranged from 0.12 to 0.18 and were higher than the heritability based on the RM (0.08). Posterior means of the genetic correlations between litter sizes of different parities were positive and ranged from 0.24 to 0.71. Data sets were simulated based on the same pedigree structure and genetic parameters of the Mule ewe population obtained from both models. The simulation showed that the relative loss in accuracy and increase in mean squared error (MSE) was substantially higher when using the RM, given that the parameters estimated from the observed data using the opposite model are the true parameters. In contrast, Bayesian information criterion (BIC) selected the RM as most appropriate model given the data because of substantial penalty for the higher number of parameters to be estimated in the MTM model. In conclusion, when the relative change in accuracy and MSE is of main interest for estimation of breeding values of litter size of different parities, the MTM is recommended for the given population. When reduction in risk of using the wrong model is the main aim, the BIC suggest that the RM is the most appropriate model.     

Use of a Piezo Drill for Intracytoplasmic Sperm Injection into Cattle Oocytes Activated with Ionomycin Associated with Roscovitine

Intracytoplasmic sperm injection (ICSI) consists of the introduction, by micromanipulation, of a single sperm into the cytoplasm of a mature egg. This technique is particularly advantageous when only a few sperm are available for fertilization, representing an important tool in preserving genetic material, especially from poorly fertile males. The results from ICSI in cattle are very often unsatisfactory and difficult to reproduce. Thus, the goal of this study was to evaluate the effect of the use of a Piezo drill (PD) and oocyte activation with ionomycin + roscovitine (I + R) during ICSI in cattle oocytes. After in vitro maturation (24 h), cumulus complex oocytes were divided into four groups: G1 – the ICSI was performed without the use of a PD and the oocyte was activated with I + R; G2 – the ICSI was performed with the use of the PD and activation with I + R; G3 – the ICSI was performed with the use of the PD, but without activation and G4 – parthenogenetic control, treated with I + R, but without sperm injection. The presumptive zygotes were cultured for 7 days and evaluated on day 3 for cleavage rate and on day 7 for blastocyst formation. Embryo production by standard in vitro fertilization in the laboratory was 78% for cleavage (117/150) and 35% for blastocyst formation (41/150). The cleavage rates obtained in G1, G2 and G4 were similar (66.7%, 71.6% and 66.3%, respectively), demonstrating the beneficial effect of oocyte activation. However, in G3, despite the presence of the sperm and the electric stimulation of a PD, the cleavage rates were significantly lower (17.5%) compared with the groups that used chemical activation, even in the absence of sperm (G4). Despite the beneficial effects of activation, this stimulus alone, or in the absence of the PD, was not sufficient for adequate morulae formation (13.4%, 37.9%, 0.0% and 13.5% for G1, G2, G3 and G4, respectively). Only in G2, when the PD was used followed by artificial activation, blastocysts were obtained (14.7%). These results indicate that cattle oocytes must be activated after ICSI to produce viable embryos.     

Experimental infection of sheep with bovine herpesvirus type-5 (BHV-5): acute and latent infection

We demonstrated that sheep are susceptible to acute and latent infection by bovine herpesvirus type-5 (BHV-5). Lambs inoculated intranasally with two South American BHV-5 isolates replicated the virus with titers up to 10(7.1) TCID50/ml for up to 15 days and showed mild signs of rhinitis. Four lambs in contact with the inoculated animals acquired the infection and excreted virus for up to seven days. One lamb developed progressive signs of neurological disease and was euthanized in extremis. Clinical signs consisted of tremors of the face, bruxism, ptyalism, incoordination, lateral flexion of the neck and head, circling, walking backwards, recumbency and paddling. The virus was detected in the anterior and posterior cerebrum, dorso- and ventro-lateral cortex, cerebellum, pons, midbrain and olfactory bulb. Viral nucleic acids were demonstrated in neurons and astrocytes of the anterior and ventro-lateral cortex by in situ hybridization. Histological changes consisting of non-suppurative meningitis, perivascular mononuclear cuffing, focal gliosis, neuronal necrosis and intranuclear inclusions were observed in the anterior cerebrum, ventro-lateral cortex and midbrain. Dexamethasone treatment at Day 50 pi resulted in reactivation of the latent infection and virus shedding in 13/16 (81%) of the lambs. Together with previous reports of BHV-5 antibodies in sheep, these findings show that sheep are fully susceptible to BHV-5 suggesting that infection by BHV-5 in sheep may occur naturally.