In vivo characterisation of two Australian isolates of Marek's disease virus including pathology, viral load and neuropathotyping based on clinical signs.

OBJECTIVE: To evaluate the pathogenicity of Australian Marek's disease virus (MDV) isolate MPF23 (1985) against the reference strain MPF57 based on pathology, viral load and neuropathotyping on the basis of clinical signs. PROCEDURE: Two MDV challenge isolates (MPF57 or MPF23) were administered to unvaccinated specific-pathogen free (SPF) layer chicks on day 5 after hatch at three challenge doses (500, 2000 or 8000 plaque-forming units (pfu)/chick). Mortality, body weight, immune organ weights, MDV load in peripheral blood lymphocytes (PBL) and clinical signs were measured to 56 days post challenge (dpc). RESULTS: MPF23 was the more pathogenic of the two viruses, inducing higher mortality (81% vs 62%) and incidence of MD lesions (100% vs 76%). MPF23 induced earlier, more sustained and more severe neurological signs in the period 26-56 dpc. However, there were few differences during the 0-23 dpc used in the neuropathotyping classification under test. The observed pattern during this earlier period classified both viruses as neuropathotype B, consistent with a very virulent pathotype. MDV load in PBL at 7 and 44 dpc did not differ between virus isolates, but the load at 7 dpc was significantly and negatively associated with time to euthanasia or death. CONCLUSION: MPF23 appears to be as, or more, virulent than the MDV strains isolated over the subsequent two decades. The neuropathotyping system developed in the USA did not clearly differentiate between the two isolates under test; however, extension of the period of assessment of clinical signs beyond 26 dpc did reveal clear differences.

Flaxseed oil supplementation alters the expression of inflammatory-related genes in dogs.

Long chain n-3 fatty acids are beneficial to mammals because of their anti-inflammatory role. However, whether flaxseed oil, which is rich in short chain n-3 fatty acids, has such a role, it has not been extensively examined. This study investigated the supplementation of flaxseed oil on the regulation of genes involved in inflammatory responses such as heat shock proteins (HSP90 and HSP70) and interleukin (IL1ß) in the white blood cells of dogs. Five beagles and 5 greyhounds were supplemented with Melrose(®) flaxseed oil at the rate of 100 mL/kg food for 21 days. The blood was collected at day 0, 15, and 22 following supplementation. The expression of 3 genes was quantified using real-time polymerase chain reaction. Plasma concentrations of fatty acids such as alpha linolenic acid, eicosapentaenoic acid, docosahexaenoic acid, linoleic acid, and arachidonic acid were measured, and their correlations with changes in gene expression were determined. Flaxseed oil supplementation downregulated the expression of HSP90 and IL1ßin greyhounds but showed no significant effect on these genes in beagles. HSP70 remained unchanged in both breeds following the supplementation. Correlations of HSP90 and IL1ßexpression levels with the plasma fatty acid concentrations on day 22 showed a significant negative correlation in greyhounds. Dietary flaxseed oil altered the expression of genes involved in inflammation in white blood cells. Because the expression of the genes may vary in different breeds, it will be useful to consider breed responses to dietary manipulation in canine nutrition management.

Oxalate nephropathy in a laboratory colony of common marmoset monkeys (Callithrix jacchus) following the ingestion of Eucalyptus viminalis.

Seven common marmoset monkeys (Callithrix jacchus) from a laboratory colony of 17 died over a period of eight months. Death of six of these monkeys was attributed to kidney failure from an oxalate-induced nephropathy. The epidemiology of this outbreak suggested an exogenous source and there was strong evidence that the source was bark and leaves from an Eucalyptus viminalis tree. Branches of this tree were introduced one month before the first death. The branches were removed one month after deaths commenced, but deaths continued for another five months. Urinalysis of all surviving marmosets at 80 and 122 days after initial contact with the E viminalis branches suggested that these monkeys had renal impairment. In the cases described here, the eating behaviour of common marmosets apparently exposed the animals to toxic levels of oxalate in the bark and leaves of an E viminalis tree.

Effects of the absence of protozoa from birth or from weaning on the growth and methane production of lambs.

Merino ewes (n 108) joined to a single sire were allocated into three flocks, with ewes in one flock being chemically defaunated in the second month of gestation. Single lambs born to defaunated ewes (BF lambs) were heavier at birth and at weaning than lambs born to faunated ewes (F lambs). After weaning, all BF and F lambs were individually housed then half of the F lambs were chemically defaunated (DF lambs). In trial 1, BF, DF and F lambs were offered a concentrate-based diet containing either 14 or 19 % protein for a 10-week period. Wool growth rate of BF lambs was 10 % higher than that of DF or F lambs and was increased 9 % by the high-protein diet. While there was no main effect of protozoa treatment on enteric methane production, there was an interaction between protozoa treatment and diet for methane production. BF and DF lambs produced more methane than F lambs when fed the low-protein diet but when fed the high-protein diet, emissions were less than (BF lambs) or not different from (DF lambs) emissions from F lambs. In trial 2, lambs were offered 800 g roughage per d and, again, methane production was not affected by the presence of protozoa in the rumen. The data indicate that while lambs without rumen protozoa have greater protein availability than do faunated ruminants, there is no main effect of rumen protozoa on enteric methane production by lambs fed either a concentrate or roughage diet.

Salmonella and on-farm risk factors in healthy slaughter-age cattle and sheep in eastern Australia.

OBJECTIVE: To examine healthy slaughter-age cattle and sheep on-farm for the excretion of Salmonella serovars in faeces and to identify possible risk factors using a questionnaire. PROCEDURE: The study involved 215 herds and flocks in the four eastern states of Australia, 56 with prior history of salmonellosis. Production systems examined included pasture beef cattle, feedlot beef cattle, dairy cattle, prime lambs and mutton sheep and animals were all at slaughter age. From each herd or flock, 25 animals were sampled and the samples pooled for Salmonella culture. All Salmonella isolated were serotyped and any Salmonella Typhimurium isolates were phage typed. Questionnaires on each production system, prepared in Epi Info 6.04, were designed to identify risk factors associated with Salmonella spp excretion, with separate questionnaires designed for each production system. RESULTS: Salmonellae were identified in all production systems and were more commonly isolated from dairies and beef feedlots than other systems. Statistical analysis revealed that dairy cattle were significantly more likely to shed Salmonella in faeces than pasture beef cattle, mutton sheep and prime lambs (P<0.05). A wide diversity of Salmonella serovars, all of which have been isolated from humans in Australia, was identified in both cattle and sheep. Analysis of the questionnaires showed access to new arrivals was a significant risk factor for Salmonella excretion on dairy properties. For beef feedlots, the presence of large numbers of flies in the feedlot pens or around stored manure were significant risk factors for Salmonella excretion. CONCLUSION: Dairy cattle pose the highest risk of all the slaughter-age animals tested. Some of the identified risk factors can be overcome by improved management practices, especially in relation to hygiene.

Salmonella Typhimurium persistence in a Hunter Valley dairy herd.

OBJECTIVE: An epidemiological study was undertaken at a Hunter Valley dairy with persistent Salmonella Typhimurium infection. The aim of the study was to identify cattle currently or previously infected with Salmonella, possible sources of the organism, patterns of spread, and husbandry practices that could be improved. METHODOLOGY: Faecal samples, feed, water and environmental samples were cultured for Salmonella and blood samples were tested for antibodies against Salmonella (Dublin and Typhimurium). A questionnaire was designed to identify possible risk factors associated with Salmonella excretion. RESULTS: S Typhimurium was apparently introduced from an old to a new dairy through manure spread as fertiliser. Salmonella apparently persisted in the effluent pond, and the following year clinical cases occurred after pasture, irrigated with water from the pond, was grazed by dry cows, and adult cattle became clinically ill with salmonellosis. The disease spread to other cows and calves. Poor design of calf pens assisted spread of Salmonella from sick to healthy calves. In addition, there was suspected transmission to the dairy farmer's 9-month-old daughter. Salmonellosis on a farm is a potential zoonotic risk to farm workers and their families. There is also the risk that cull cows may carry Salmonella to the abattoir and subsequently into the human food chain. Methods of waste management, and the design of calf pens, were identified as major risk factors that could be improved to minimise the spread of salmonellosis on this property.

A lack of predatory interaction between rumen ciliate protozoa and Shiga-toxin producing Escherichia coli.

AIMS: To investigate interactions between rumen protozoa and Shiga toxin-producing Escherichia coli (STEC) and to ascertain whether it is likely that rumen protozoa act as ruminant hosts for STEC. METHODS AND RESULTS: The presence of stx genes in different microbial fractions recovered from cattle and sheep rumen contents and faeces was examined using PCR. In animals shedding faecal STEC, stx genes were not detected in the rumen bacterial or rumen protozoal fractions. Direct interactions between ruminal protozoa and STEC were investigated by in vitro co-incubation. Rumen protozoa did not appear to ingest STEC, a STEC lysogen or non-STEC E. coli populations when co-incubated. CONCLUSIONS: The ruminal environment is unlikely to be a preferred habitat for STEC. Bacterial grazing by rumen protozoa appears to have little, if any, effect on STEC populations. SIGNIFICANCE AND IMPACT OF THE STUDY: This study indicates that ruminal protozoa are unlikely to be a major factor in the survival of STEC in ruminants. They appear as neither a host that protects STEC from the ruminal environment nor a predator that might reduce STEC numbers.

The efficacy of an albendazole-medicated block in controlling sheep nematodes in Xinjiang province, north-west China.

Medicated feed-blocks containing 2 mg/kg albendazole were fed for 12 days to ewe-lambs naturally infected with nematode worms in north-west China. The cumulative total average intake of albendazole was 15 mg/kg body weight. This treatment was compared with a single oral drench of 15 mg/kg albendazole and with no treatment. Both albendazole treatments were 99% effective in reducing the total strongyle egg counts. The medicated block was 100% effective in reducing the total trichostrongylid worm count, although only 86.4% of the sheep consumed an adequate amount. The single drench was 97% effective. Under the management conditions available to sheep herders in the northern provinces of China, albendazole-medicated feed blocks used at strategic times could effectively control trichostrongylid nematodes.

Field trials of ephemeral fever vaccines.

Various bovine ephemeral fever (BEF) vaccines were tested between 1982 and 1984 in 24 Queensland herds; neutralising antibody responses were monitored and six of the herds were closely observed following natural challenge with ephemeral fever. A vaccine regime of two consecutive vaccinations with attenuated virus combined with the adjuvant Quil A provided excellent protection against BEF for at least 12 months, whereas one vaccination with the Quil A vaccine or two vaccinations with vaccine containing the adjuvant aluminium hydroxide gel did not provide significant protection. Antibody responses were highest for the vaccine incorporating Quil A when it was given as two consecutive injections. There was an apparent relationship between neutralising antibody response and the level of protection.

Detection of papillomavirus DNA in precancerous lesions of the ears of sheep.

Sixty-seven benign precancerous cutaneous lesions from the ears of 51 sheep were examined for papillomavirus DNA by hybridisation to radioactively labelled or biotinylated probes of bovine papillomavirus type 1 (BPV 1) DNA under varying conditions of stringency. An additional 16 precancerous lesions from other cutaneous sites on 15 sheep and 15 samples of lesion-free skin from nine sheep were similarly examined. Both total genomic and subgenomic probes were used. DNA from 10 aural lesions and one vulval lesion reacted with the probe in a manner indicative of the presence of episomal papillomavirus DNA. Papillomavirus DNA was detected at low stringency in eight of the 10 aural lesions and the vulval lesions, and at high stringency in two aural lesions. Three out of the 8 aural lesions that were positive at low stringency reacted when re-tested at high stringency. Hybridisation with one of the samples of lesion-free ovine skin produced occasional equivocal signals. One particular positive lesion, an ovine aural cutaneous horn, was studied in more detail. When treated with restriction endonucleases, its restriction enzyme pattern was the same as that for BPV 2 DNA with eight of twelve enzymes and the same as that for BPV 1 DNA with two of the twelve enzymes. It was concluded that this ovine papillomavirus was more closely related to BPV 2 than to BPV 1. The possibility that it could be a subtype of BPV 2 is discussed.

BCG emulsion immunotherapy of equine sarcoid.

Of 61 horses with sarcoids treated with intralesional injection of a double emulsion incorporating inactivated bacillus Calmette Guérin organisms, 36 (59 per cent) showed complete regression and 11 (18 per cent) showed partial regression. The majority of cases required only one treatment. Not all sarcoids were responsive to this therapy; those not responding were usually large or on horses with multiple sarcoids.

A bovine ephemeral fever vaccine incorporating adjuvant Quil A: a comparative study using adjuvants Quil A, aluminium hydroxide gel and dextran sulphate.

Various vaccines containing the 919 strain of ephemeral fever virus were evaluated in experimental calves and in commercial cattle. The vaccine virus was mixed with one of the adjuvants, Quil A (a saponin derivative), aluminium hydroxide gel, dextran sulphate or combinations of these. The response of experimental calves was evaluated by measuring the production of neutralising antibodies and by resistance to challenge with virulent virus; the response of commercial cattle was judged only by the production of neutralising antibody. Twelve calves given two doses of vaccine containing Quil A produced neutralising antibodies to bovine ephemeral fever virus and all were resistant to challenge with virulent virus given 28 to 76 days after the second vaccination. The vaccine given in three of these calves also contained aluminium hydroxide gel. Six of eight unvaccinated control calves succumbed to experimental challenge. In commercial cattle (17 to 26 animals per group) the serological response after two doses of vaccine containing Quil A or Quil A and dextran sulphate was significantly better than that after vaccines containing only dextran sulphate or after vaccines containing combinations of aluminium hydroxide gel and Quil A. The adjuvant Quil A alone was tested in cattle and shown to produce a transient soft swelling at the injection site as well as a rise in rectal temperature of greater than 1 degree C one day after inoculation. At least 99.99 per cent of viral infectivity was destroyed when the vaccine was mixed with Quil A, suggesting that live virus may not be essential in the immunogenicity of the vaccine. This vaccine overcame two of the problems associated with previous attenuated vaccines tested in Australia; the necessity for adjuvant and virus to be mixed immediately before use and the large volume of the vaccine.

An epizootic of bovine malignant catarrh in Malaysia.

A severe epizootic of bovine malignant catarrh occurred from November 1976 until June 1977 in cattle at an agricultural institute in peninsular Malaysia. In a group of 82 Kedah-Kelantan cattle the morbidity rate was 47.6 per cent with a fatality rate of 89.7 per cent. In a group of 43 local Indian dairy cattle the morbidity rate was 23.3 per cent with a fatality rate of 100 per cent. Although evidence suggested that sheep acted as a common source of infection, the disease occurred in one animal which had no contact with sheep but had contact with infected cattle and carcases.