Development and evaluation of a rapid absorbed enzyme immunoassay test for the diagnosis of Johne''s disease in cattle

An absorbed enzyme immunoassay (EIA) test for Johne's disease in cattle was developed in which absorption of cross-reacting antibodies occurred as a rapid reaction in solution rather than overnight with whole organisms and a subsequent centrifugation step. Total test time was reduced to less than 2 h with a minimum of manipulations. The test was evaluated in cattle herds from Johne's disease-endemic and Johne's disease-free regions of Australia. Specificity was 99.8%. Calculations of sensitivity were affected by the history of the herd under test. However, the EIA detected in excess of 80% of animals before onset of clinical disease and 65% of faecal shedders were EIA positive on, or before, first detection of Mycobacterium paratuberculosis in their faeces. The test should aid epidemiological studies and be a useful tool in the management and control of Johne's disease.     

The prevalence of itchmite, Psorergates ovis, among sheep flocks with a history of fleece derangement

An investigation of sheep flocks in the main sheep raising areas of New South Wales showed that the itchmite Psorergates ovis was frequently associated with fleece derangement. In 26 of the 41 flocks examined, P. ovis was the only ectoparasite detected. P. ovis and the sheep body louse Damalinia ovis, were found in 5 flocks. No external parasites were found on sheep examined from the 10 remaining flocks. The type of fleece derangement most frequently recorded was rubbing which in some cases was combined with areas of chewed fleece. Among flocks, there were positive relations between the prevalence of fleece derangement and prevalence of itchmite or scurf and between itchmite count and mean scurf score. Within flocks, itchmite infested sheep or sheep with scurf had higher prevalences of fleece derangement than sheep on which no mites or no scurf were found. Itchmite infested sheep had a higher prevalence of scurf than those with no detectable mite infestation. There were no significant differences in itchmite populations or fleece derangement between untreated flocks and flocks treated with synthetic pyrethroids, organophosphates or arsenic and rotenone.     

Comparison of inoculation regimes for the experimental production of swine erysipelas arthritis: II Serological findings in a gel diffusion precipitin test and enzyme-linked immunosorbent assay

The serological response of pigs to Erysipelothrix rhusiopathiae inoculation was monitored by a gel diffusion precipitin test (GDPT) using a crude, serotype-specific, autoclaved antigen and an enzyme-linked immunosorbent assay (ELISA) using a heat-extracted, alcohol precipitated and molecular seived antigen previously shown to react with serum from pigs infected with serotypes 1 or 2. All pigs receiving 3 or 5 weekly intravenous inoculations of either a highly virulent (VRS 229) or a lowly virulent isolate (VRS 252) produced GDPT-reactive antibody within 3 weeks, but only 44% were still reactive at 8 to 9.5 weeks. The ELISA response was significantly higher in pigs inoculated with the highly virulent strain, and was similar in pigs receiving 3 or 5 doses of either strain. In a dose-response trial, after 3 doses of VRS 229, GDPT reactivity occurred earlier and was stronger in pigs given higher doses of E. rhusiopathiae, but the response peaked 3 to 5 weeks after the start of challenge and was short lived. GDPT reactivity correlated with dose, but not with the severity of arthritis. The ELISA demonstrated specific IgG antibody was present by 2 weeks, and persisted to at least 11 weeks. The ELISA reactivity was significantly higher in pigs with arthritis than in pigs that received low doses and were not arthritic. Within groups of pigs with arthritis a significant, dose dependent, linear ELISA response developed but did not correlate with the presence or degree of arthritis at slaughter. Non-arthritic pigs had similar low ELISA responses to uninoculated controls.     

Relationships Between Fertility and Some Parameters in Male Broiler Breeders (Body and Testicular Weight,Histology and Immunohistochemistry of Testes,Spermatogenesis and Hormonal Levels)

In a farm of grandparent broiler breeder chickens, we followed the development of 350 roosters from 6 to 55 weeks of age. Data collected and evaluated from these males included body weight, testicular weight, histologic and immunohistochemistry studies of the testes, hormone levels (testosterone, estradiol and corticosterone) and sperm production. The objective was to understand the factors that affect or influence hatch loss that is commonly observed after 45 weeks of age in breeder flocksare often correlated to broiler breeder male chickens. The results of this study showed that in conjunction with the weight of the rooster, the testicle weight increases quickly after the rooster receives light stimulation. At an older age, the study showed that there is a process of testicular shrinkage, and the same effect is seen in sperm production and testosterone levels in broiler breeder roosters. From the histology evaluation, we defined 5 histologic phases that illustrate the evolution of the testicular tissue: perinatal, infantile, puberty, adult and senile. We observed that the adult males with a body weight <3800 g were infertile or had subfertile levels and also had low levels of testosterone and high levels of corticosterone. In contrast, the heaviest males showed correct testicular vitality, high levels of testosterone and low levels of corticosterone. However, the roosters that had acquired this high body weight were also at risk of having less complete copulations because of their physical mass. The loss of uniformity of the males and the appearance of hierarchies within the flock accompany a decline in the percentages of hatches as a consequence of the poor confirmation of the males for copulation or the restriction to the access to the females. Results of this study show that the decrease in fertility from 45 weeks of age has been associated with a decline in testicular weight, sperm production and the testosterone levels in animals with a sub‐par weight. Likewise, decreasing hatch in older flocks may also result from a loss of conformation, and the lack of complete copulations is possible because of animals that are grossly overweight.     

A preliminary survey of Chlamydia psittaci genotypes from native and introduced birds in New Zealand

AIM: To describe the Chlamydia psittaci genotypes in samples from native and introduced birds from New Zealand by analysis of the sequence variation of the ompA gene.

METHODS: DNA was extracted from samples collected from a non-random sample of birds; either swabs from live asymptomatic birds or birds with clinical signs, or formalin-fixed, paraffin-embedded (FFPE) samples from historical post-mortem cases. The presence of C. psittaci in all samples had been confirmed using a quantitative PCR assay. The C. psittaci ompA gene was amplified and sequenced from samples from 26 native and introduced infected birds comprising 12 different species. These sequences were compared to published available C. psittaci genotypes.

RESULTS: Genotypes A and C of C. psittaci were identified in the samples. Genotype A was identified in samples from nine birds, including various native and introduced species. Genotype C was identified in samples from 16 different waterfowl species, and a mixed infection of both genotypes was found in a kaka (Nestor meridionalis). In native birds, C. psittaci infection was confirmed in seven new host species.

CONCLUSIONS AND CLINICAL RELEVANCE: Two genotypes (A and C) of C. psittaci were found in samples from a wider range of both native and introduced species of birds in New Zealand than previously reported. Both genotypes have been globally associated with significant disease in birds and humans. These initial results suggest the host range of C. psittaci in New Zealand birds is under-reported. However, the prevalence of C. psittaci infection in New Zealand, and the associated impact on avian and public health, remains to be determined. There are biosecurity implications associated with the importation of birds to New Zealand if there is a limited diversity of C. psittaci genotypes present.     

Pathological Conditions of the Reproductive Organs of Male Stray Dogs in the Tropics: Prevalence, Risk Factors, Morphological Findings and Testosterone Concentrations

The objective of this study was to estimate the prevalence of and risk factors for pathological conditions of the reproductive organs in stray dogs under tropical conditions. Three hundred and eighteen dogs were examined post-mortem in the period from 1 July 2002 to 30 June 2003. Before killing, a blood sample (from the cephalic vein) for testosterone assay was taken. Pathological conditions of the reproductive organs were found in 135 of the dogs (42.5%) and in 175 of the testes (64.8%). The most frequent pathologies found were testicular degeneration, cryptorchidism, testicular hypoplasia and testicular tumours (in 15.1%, 6.6%, 6.6% and 5.4% of the dogs and 15.1, 4.6, 6.0 and 3.5 of the testes, respectively). Transmissible venereal tumour (TVT) was seen in 5.4% of the dogs. Testicular degeneration was more common in old dogs and underweight dogs (p < 0.05). Testicular tumours were 14.3 times more common in cryptorchid dogs. Age was another important factor for the development of testicular tumours (p < 0.05). Lower levels of testosterone concentration (p < 0.05) were observed in dogs with advanced testicular degeneration (0.7 +/- 0.8 nM), dogs with hypoplastic testicles (0.8 +/- 0.9 nM) and dogs with one degenerated and one retained testis or with bilateral cryptorchidism (1.2 +/- 0.9 nM) compared to dogs with one or two normal testes (7.0 +/- 5.5 nM). Testicular volume and weight were significantly lower in degenerated, hypoplastic and retained testes compared with the contralateral normal testis. Some spermatogenic activity was found in three of the retained testes, producing oligozoospermic smears with a high percentage of sperm abnormalities. No comparable epidemiological data about male pathological conditions of the reproductive organs in the dog is available. The prevalence found in this study, yet, appears high.     

Clinical evaluation of protein electrophoresis of cerebral spinal fluid

Protein fractions of cerebral spinal fluid samples were analyzed in an effort to find a reliable, minimally invasive and cost effective method of diagnosing cervical vertebral myelopathy. There were six separate protein fractions analyzed by electrophoresis. In this study we specifically looked for a post beta spike in horses with spinal cord compression. There was no correlation in our study between horses with a post beta spike and CVM as diagnosed by a myelogram.