An estimate of specificity for a Johne's disease absorbed ELISA in northern Australian cattle

Objective To estimate the specificity of an absorbed enzyme-linked immuno-sorbent assay kit^d for Johne's disease (JD) when used in mature cattle populations resident in northern Australia.
Design Blood samples were collected from beef cattle in northern Queensland, the Northern Territory and northern Western Australia, and from dairy cattle in northern Queensland. The specificity of a serological test for JD was estimated by testing the blood samples with an absorbed ELISA kit. Further samples were collected from cattle with positive ELISA results to determine the presence or absence of infection with Mycobacterium avium subsp paratuberculosis .
Procedure During 1995 and 1996, blood, tissue and gut contents were collected from beef cattle at abattoirs in Queensland and the Northern Territory; and blood and faecal samples were collected from dairy cattle in herds assessed to be most at risk for JD in northern Queensland. The blood samples were tested using an absorbed ELISA kit. Tissues and gut contents from beef cattle that had positive ELISA results were cultured for M avium subsp paratuberculosis , and tissues were examined histo-logically. Faecal samples from dairy cattle with positive ELISA results were cultured for M avium subsp paratuberculosis .
Results Estimates of specificity for this absorbed ELISA in mature northern Australian cattle were 98.0% (97.0 to 98.8%, 95% CI) in beef cattle, and 98.3% (96.7 to 99.3%, 95% CI) in dairy cattle.
Conclusion Estimates of specificity in this study were lower for beef cattle from the Northern Territory and northern Western Australia and for dairy cattle from northern Queensland than those quoted from studies on cattle in southern Western Australia. This should be considered when serological testing using the JD ELISA is carried out on northern Australian cattle.     

The effects of furosemide and pentoxifylline on the flow properties of equine erythrocytes:In vitro studies

The effects of various concentrations of furosemide and pentoxifylline on equine RBCin vitro were evaluated to facilitate better understanding of the potential effects of these drugs on blood flow properties. Furosemide induced increased mean cell volume (MCV), increased RBC potassium concentration, increased whole blood viscosity, and decreased the RBC filtrability. These data indicate that furosemide may block the RBC membrane transport pathways resulting in potassium and water retention. The increase in size and the resultant decrease in the surface-area-to-volume ratio may have caused the impaired RBC filtrability and increased blood viscosity. Pentoxifylline improved RBC filtrability without changing the RBC size or the potassium or chloride concentrations, suggesting that pentoxifylline may increase the deformability of the RBC membrane. The study indicated that pentoxifylline has potential therapeutic applications for improving microvascular blood flow but that furosemide may have adverse effects on blood flow.Abbreviations cP centipoise - EIPH exercise-induced pulmonary haemorrhage - MCV mean cell volume - Na-K-2Cl sodium potassium 2-chloride - PCV packed cell volume - RBC red blood cell     

Prekallikrein deficiency in a family of Belgian horses

A 7-year-old Belgian stallion hemorrhaged excessively after castration; the hemostatic mechanism was investigated. The horse had normal one-stage prothrombin time and markedly prolonged activated partial thromboplastin time (APTT). Results of intrinsic coagulation factor assays were all normal with the exception of prekallikrein activity, which was markedly reduced (less than 1% activity; value for control population, 63 to 150%). Two of this horse's full siblings, a brother and sister, had markedly prolonged APTT and low prekallikrein values (2.5% and less than 1%, respectively). The addition of plasma from a normal equine plasma pool corrected the prolonged APTT in the 3 Belgian sibling with low prekallikrein activity. Prekallikrein activity in 10 other closely related Belgian horses ranged between 12.5 and 64% (mean, 29.3%), compared with 63 to 150% (mean, 91%) in 10 mixed-breed horses. In the 3 Belgian siblings with low prekallikrein activity, the APTT approached normal after prolonged incubation (15 minutes) with the contact activator and in response to addition of an ellagic acid activator. The 3 Belgian siblings with low prekallikrein activity may be homozygous for prekallikrein deficiency, whereas the other close relatives may be heterozygous for the genetic defect.     

Comparison of the Johne''s absorbed EIA and the complement-fixation test for the diagnosis of Johne''s disease in cattle

A commercially available absorbed ELISA for the diagnosis of Johne's disease (JD) (paratuberculosis) in cattle, the Johne's Absorbed EIA, was compared with the conventional complement-fixation test (CFT) used in Australia. Stored plasma from 3 Victorian dairy herds with a history of JD, sera from specimens submitted from animals showing clinical signs of JD and sera from the US National Repository for Paratuberculosis Specimens were used to determine the sensitivity of each test. The EIA detected 48.8% of 43 Australian animals with subclinical JD, while the CFT detected only 12 (21.4%) of 56 subclinically affected cattle. Of 150 subclinically infected US cattle, the EIA detected 47.3% and the CFT detected 52.0%. The EIA detected 59.7% of animals which at the time of sampling were shedding Mycobacterium paratuberculosis in their faeces, but showed no clinical signs of JD, while the CFT detected 57.3%. The EIA correctly identified 88.2% of 136 histologically confirmed clinical cases, and the CFT detected 83.4%. The specificity of each test was determined by testing sera collected at slaughter from animals residing in a known JD-free area of Australia, and from samples from the US National Repository of Paratuberculosis Specimens collected from certified-free herds in Wisconsin. The EIA was found to have a specificity of 99.8% when 998 Australian animals were used as the test population, and 99.0% when 196 US animals were used. The specificity of the CFT using Australian samples was 96.9% and 95.2% using American samples.     

Functional and morphological studies on blood platelets in a thrombasthenic horse

A four-year-old Standardbred gelding presented with a 3.5 year history of intermittent epistaxis and spontaneous submucosal petechiae and ecchymoses in the nares and the mouth. Routine haematological and biochemical examinations were unremarkable. A thrombocytopathy was suspected when activated partial thromboplastin time, one stage prothrombin time, plasma fibrinogen and the platelet count were all normal. The patient's platelets failed to aggregate with serotonin, adenosine diphosphate, collagen (at 20 micrograms/ml) or the endoperoxide analogue U46619. Very high levels of collagen (100 micrograms/ml) did cause aggregation. The response to the calcium ionophore A23187 was reduced and although complete degranulation occurred the resulting aggregates were unstable. Thromboxane generation in response to collagen and ADP was inferred from the concentration of its stable metabolite thromboxane B2 and was reduced. A diagnosis of a thrombasthenia-like syndrome possibly equivalent to Type II Glanzmann's thrombasthenia in people was made.     

Isolation and pathogenicity of Australian strains of Eimeria praecox and Eimeria mitis

Objective To determine the presence of E praecox and E mitis in Australia, to isolate representative strains of these species from chickens and determine their pathogenicity.
Design Morphological, physiological and cross protection studies were undertaken to confirm the identity of Australian isolates of E praecox and E mitis.
Procedure Oocysts were isolated from a backyard flock at Jimboomba, southeastern Queensland and numbers of E praecox and E mitis enriched by passage in chickens immune to five other species of poultry Eimeria . Oocysts of mean conformation and size of the two species were purified by single oocyst passage. Two isolates that closely matched recorded parameters for E praecox and E mitis were selected and designated JP and JM respectively. The cross protection between the isolates and E acervulina was determined by infection and challenge experiments. The virulence of the two isolates was determined by comparing weight gains of groups of birds inoculated with JP isolate or JM isolate with untreated groups.
Results Isolates JP and JM most closely matched recorded parameters of E praecox and E mitis respectively. Groups of chickens, previously infected with JP and JM isolates, showed no significant protection against infection with E acervulina . In a separate trial, groups of susceptible chickens inoculated with 10⁵ oocysts of JP and JM isolates showed significantly reduced weight gains compared with untreated controls.
Conclusion Isolates JP and JM are E praecox and E mitis respectively, confirming the presence of these species in Australia. These isolates were found capable of causing significant reductions in weight gains in susceptible chickens.     

Use of enzyme-linked immunoassays for antibody to types C and D botulinum toxins for investigations of botulism in cattle

The development of specific enzyme-linked immunosorbent assays (ELISA) for antibody to types C and D Clostridium botulinum toxins for investigation of botulism in cattle is described. Partially purified type C and D toxins were used as antigens to develop these ELISAs. Specificity of the ELISAs was evaluated on sera from 333 adult beef and dairy cattle from areas with no history or evidence of botulism in animals or water birds. The test was also evaluated on sera from 41 herds that included herds vaccinated against botulism, confirmed botulism cases and herds from areas where the disease is considered endemic. The ELISAs detected the presence of antibody to botulinum toxins in samples from vaccinated cattle and both convalescent and clinically normal animals from unvaccinated herds with outbreaks of botulism. Antibody was also found in unvaccinated animals from herds in which there had been no diagnosed botulism cases in areas where botulism was considered endemic. Sera from some unvaccinated cattle with high ELISA reactivity was shown to be protective for mice in botulinum toxin neutralisation tests. The use of these tests in investigations of botulism in cattle is discussed.     

Leptospira interrogans serovar hardjo is not a major cause of bovine abortion in Victoria

The aim of this study was to determine whether evidence could be obtained of foetal infection with Leptospira interrogans serovar hardjo in aborted foetuses collected from dairy farms. Material from 197 abortions occurring over a wide area of Victoria was collected over 3 years. None of 195 foetal kidney cultures or 7 cultures from membranes was positive for leptospiral organisms. Immunogold silver staining for leptospires was performed on sections of kidneys, lungs or heart from 156 foetuses, with negative results. Evidence of transient leptospiral infection in 11 of 123 foetuses was obtained by foetal heart blood serology. Two isolates of L. interrogans serovar hardjo were obtained from the urine of milking cows. These strains were examined by restriction endonuclease analysis and both were shown to be of the genotype Hardjobovis, as have been all Australian isolates studied so far. It appears that foetal infection with serovar hardjo is not associated with any substantial proportion of bovine abortions in Victoria, in contrast to the situation in Northern Ireland. The apparent absence from Victoria of the pathogenic genotype Hardjoprajitno is a possible explanation.     

The survival of platypuses in captivity

Data are presented on the duration of survival of 228 platypuses at six Australian zoos between 1934 and 1988. Only 22.4% of all platypuses survived more than 1 year in captivity. Of 15 living platypuses, 3 had been held in captivity for less than 1 year, 5 for between 1 and 5 years, 6 for between 5 and 10 years and 1 for 21 years. Of 213 platypuses that died in captivity, 81.7% had died within 1 year; most within the first month. The duration of survival was unrelated to the age of animals at acquisition or to sex. The survival rate of animals donated to zoos, including "refugees", was similar to that of purpose-caught animals. Clearly, only a small proportion of platypuses adapted to captive husbandry. The cause of death of most platypuses was not established. However, infectious disease did not appear to be significant. Approximately 28% of deaths were related to inadequate husbandry. Recommendations are made to improve the survival of platypuses in captivity. Research has commenced in zoos to facilitate this goal.