Bluetongue disease in sheep in New South Wales - April 2023.

In 2016, bluetongue virus (BTV), serotype 16 (BTV-16), was detected in New South Wales (NSW) in sentinel cattle for the first time. Over the next 6 years, BTV-16 has been detected regularly and over an increasing area of the BTV zone in NSW. In April 2023, disease was reported in sheep on two farms on the Northern Tablelands of NSW. The consistent clinical signs included reduced exercise tolerance, facial swelling, serous nasal discharges with encrustation of the nasal plane, subcutaneous oedema of the neck and brisket and variable congestion of the coronary band. Affected sheep were mainly mature ewes and rams, with an estimated morbidity of 20% over a period of 6-8 weeks. Although there were several unexpected deaths, no veterinary examination was sought. Predominantly BTV-16 RNA was detected in sick sheep, with an incidence of infection of approximately 40% in a cross section of one flock. These events represent the first confirmation of disease due to bluetongue virus in NSW. As these cases occurred in a region with a high density of sheep, if there is ongoing transmission of BTV-16 during subsequent summers, further disease might be expected.

The potential for bluetongue virus serotype 16 to cause disease in sheep in New South Wales, Australia.

BLUETONGUE VIRUS SEROTYPE 16 DETECTION IN NSW: In coastal New South Wales (NSW), bluetongue virus (BTV) serotypes 1 and 21 are endemic and transmitted in most years without evidence of disease. However, serotype 16 (BTV-16) infection was detected for the first time in NSW in November 2016 in cattle undergoing testing for export. Retrospective testing of blood samples collected from sentinel cattle as part of the National Arbovirus Monitoring Program (NAMP) established that the first detected transmission of BTV-16 in NSW occurred in April 2016 in sentinel cattle on the NSW North Coast. Subsequently, until 2022, BTV-16 has been transmitted in most years and was the predominant serotype in the 2018-2019 transmission season. The data available suggests that BTV-16 may have become endemic in NSW. EXPERIMENTAL STUDIES: During experimental infection studies with BTV-16, all sheep were febrile, with the peak of viremia occurring 6-10 days after inoculation. There was nasal and oral hyperaemia in most sheep with several animals developing a nasal discharge and nasal oedema. All sheep developed coronitis of varying severity, with most also developing haemorrhages along the coronary band. There was a high incidence of haemorrhage in the pulmonary artery, epicardial petechiae, extensive pericardial haemorrhages and moderate body cavity effusions including pericardial effusions. CONCLUSION: Overall, experimental pathogenicity findings suggest moderate disease may occur in sheep in the field. These findings, when combined with climatic variability that could result in an expansion of the range of Culicoides brevitarsis into major sheep-producing areas of the state, suggest that there is an increasing risk of bluetongue disease in NSW.

What can we learn from over a decade of testing bats in New South Wales to exclude infection with Australian bat lyssaviruses?

Australian Bat lyssaviruses (ABLV) are known to be endemic in bats in New South Wales (NSW), Australia. These viruses pose a public health risk because they cause a fatal disease in humans that is indistinguishable from classical rabies infection. All potentially infectious contact between bats and humans, or between bats and domestic animals, should be investigated to assess the risk of virus transmission by submitting the bat for testing to exclude ABLV infection. The aim of this study was to establish the prevalence of ABLV infection in bats submitted for testing in NSW and to document any trends or changes in submission and bat details. We examined all submissions of samples for ABLV testing received by the NSW Department of Primary Industries Virology Laboratory for the 13-year period between 1 May 2008 and 30 April 2021. Fifty-four (4.9%) ABLV-infected bats were detected, with some clustering of positive results. This is greater than the prevalence estimated from wild-caught bats. All bats should be considered a potential source of ABLV. In particular, flying-foxes with rabies-like clinical signs, and with known or possible human interaction, pose the highest public health risk because they are more likely to return a positive result for ABLV infection. This review of ABLV cases in NSW will help veterinarians to recognise the clinical presentations of ABLV infection in bats and emphasises the importance of adequate rabies vaccination for veterinarians.

Encephalomyocarditis virus infection in alpacas.

Encephalomyocarditis virus (EMCV) infection was detected by real-time reverse transcription PCR (qRT-PCR) in four adult alpacas (Vicugna pacos) from two properties on the Far North Coast of New South Wales (NSW) in April and May 2018 and in two adult alpacas from a third property on the Central Coast of NSW in October 2018. Viral RNA was detected in a range of samples, including blood, fresh body organs and mucosal swabs. EMCV was isolated from the blood and body organs of five of these alpacas. These animals displayed a range of clinical signs, including inappetence, colic, recumbency and death. Necropsy findings included multifocal to coalescing areas of myocardial pallor, pulmonary congestion and oedema, hepatic congestion and serosal effusion. Histopathological changes comprised acute, multifocal myocardial degeneration and necrosis, with mild, neutrophilic and lymphocytic inflammation (5/5 hearts) and mild, perivascular neutrophilic meningoencephalitis (1/3 brains). This is the first report of disease due to EMCV in alpacas under farm conditions, and it identifies EMCV infection as a differential diagnosis for acute disease and death in this camelid species. In addition to the samples traditionally preferred for EMCV isolation (fresh heart, brain and spleen), blood samples are also appropriate for EMCV detection by qRT-PCR assay.

Clinical and epidemiological features of West Nile virus equine encephalitis in New South Wales, Australia, 2011.

BACKGROUND: Between February and June 2011, more than 300 horses with unexplained neurological disease were observed in New South Wales, Australia. A virulent strain of West Nile virus (WNVNSW2011 ), of Australian origin, was shown to be the cause of many of these cases. METHODS: We reviewed the clinical descriptions provided by veterinary practitioners and the associated laboratory results. Although there was a range of clinical signs described, ataxia was the only sign that was consistently described in laboratory-confirmed cases. RESULTS: WNV was detected in brain samples by real-time reverse transcription PCR assay and virus isolation. For serological confirmation of clinical cases, an equine IgM ELISA specific for WNV was shown to be the most effective tool. CONCLUSION: A state-wide serological survey undertaken after the outbreak indicated that, contrary to expectation, although infection had been widespread, the seroprevalence of antibodies to WNV was very low, suggesting that there could be a significant risk of future disease outbreaks.

Bungowannah virus--a probable new species of pestivirus--what have we found in the last 10 years?

Bungowannah virus was discovered following an outbreak of stillbirths and sudden death in young pigs. Affected animals consistently showed a myocardopathy with signs of cardiac failure. After virus isolation and PCR investigations were unsuccessful, direct fetal inoculation was undertaken. Nucleic acid purified from serum from infected fetuses was subjected to sequence-independent single-primer amplification and nucleic acid sequencing. Sequences consistent with a pestivirus were obtained. The entire genome was identified but was genetically remote from the recognized pestivirus species. This virus was not recognized by pan-pestivirus reactive monoclonal antibodies but was subsequently detected in cell cultures by immunoperoxidase staining using convalescent sow serum. Experimental infections of sows at different stages of gestation reproduced the myocarditis syndrome. Pre-weaning losses of 70 and 29% were observed following infection at days 35 and 90, respectively. Piglets infected at day 35 were shown to be persistently infected, while chronic infections were observed after fetal infection at day 55. Chronically infected piglets showed growth retardation and were viremic for up to 7 months. Myocarditis was associated with infection in late gestation (day 90). Non-pregnant sheep and cattle have been experimentally infected but with no evidence of disease. Infection of pregnant cattle in early gestation resulted in both maternal and fetal infection, but all infected fetuses mounted an antibody response to the virus. Analysis of the nucleic acid sequence confirmed that Bungowannah has a number of changes not observed in other pestiviruses. Genes encoding some of the structural proteins remain fully functional when inserted into a bovine viral diarrhea virus (BVDV) backbone. Cell culture-based studies have shown that Bungowannah virus will grow in cells extending from humans to bats as well as farm animals.

Genetic and antigenic characterization of Bungowannah virus, a novel pestivirus.

Bungowannah virus, a possible new species within the genus Pestivirus, has been associated with a disease syndrome in pigs characterized by myocarditis with a high incidence of stillbirths. The current analysis of the whole-genome and antigenic properties of this virus confirms its unique identity, and further suggests that this virus is both genetically and antigenically remote from previously recognized pestiviruses. There was no evidence of reactivity with monoclonal antibodies (mAbs) that are generally considered to be pan-reactive with other viruses in the genus, and there was little cross reactivity with polyclonal sera. Subsequently, a set of novel mAbs has been generated which allow detection of Bungowannah virus. The combined data provide convincing evidence that Bungowannah virus is a member of the genus Pestivirus and should be officially recognized as a novel virus species.

Application of a real-time polymerase chain reaction assay to the diagnosis of bovine ephemeral fever during an outbreak in New South Wales and northern Victoria in 2009-10.

OBJECTIVE: To report the occurrence of an epizootic of bovine ephemeral fever (BEF) in New South Wales (NSW) and northern Victoria in 2009-10 and describe the application of a real-time reverse transcription polymerase chain reaction (qRT-PCR) assay during the outbreak. PROCEDURES: Whole-blood samples from animals exhibiting clinical signs of BEF were requested from district veterinarians in NSW. In addition, samples were submitted from private practitioners in NSW and Victoria. In NSW, samples from animals showing acute clinical signs of BEF were tested using a qRT-PCR assay. Serological testing for BEF diagnosis was undertaken as required. Virus isolation was performed on selected samples in which bovine ephemeral fever virus (BEFV) RNA was detected. Archival serum samples and mosquito homogenates were also tested for BEFV by qRT-PCR. RESULTS: Accessions were received from 121 properties in NSW, with cases of BEF confirmed on 84 properties by qRT-PCR and 20 properties by serology. In northern Victoria, BEF was confirmed on 25 properties based on serological testing. Screening of samples by qRT-PCR enhanced the success of BEFV isolation. BEFV RNA was successfully detected in archival serum samples and a single mosquito homogenate. CONCLUSIONS: The 2009-10 outbreak resulted in the most extensive transmission of BEFV in NSW and Victoria since 1995-96, and follows a smaller outbreak in summer-autumn 2008. The use of qRT-PCR for BEF diagnosis offers veterinarians and cattle owners rapid confirmation of infection (1-2 days) and provides 'real-time' information about the presence of the disease in a district.

A prospective longitudinal study of naturally infected horses to evaluate the performance characteristics of rapid diagnostic tests for equine influenza virus.

An outbreak of equine influenza (EI) occurred in Australia in 2007. During the laboratory support for this outbreak, real-time reverse transcriptase polymerase chain reaction (qRT-PCR) assays and a blocking enzyme linked immunosorbent assay (bELISA) were used as testing methods to detect infection with the virus. The qRT-PCR and bELISA tests had not been used for EI diagnosis before, so it was not known how soon after infection these tests would yield positive results, or for how long these results would remain positive. To answer these questions, nasal swabs and blood samples were collected daily from a group of 36 naturally infected horses. EI viral RNA was detected in all horses by qRT-PCR from the first to tenth day after clinical signs were evident, and was detected in some horses for up to 34 days. Antibody was detected in the bELISA in some horses by day 3, with a median time to seroconversion of 5 days. The results from this study indicate that viral RNA can be detected from nasal swabs for much longer than infectious virus is thought to be shed from horses. The bELISA detected antibodies against EI virus in all horses for 139 days following infection, but only detected approximately 50% of horses 12 months following infection. Haemagglutination inhibition testing detected antibodies against H3 antigens in all horses for 28 days following infection, but 2 were negative by 35 days following infection.

Infection of dogs with equine influenza virus: evidence for transmission from horses during the Australian outbreak.

During the equine influenza (EI) outbreak, respiratory disease was observed in dogs that were in close proximity to infected horses. Investigations were undertaken to exclude influenza virus infection. Of the 23 dogs that were seropositive in tests using the influenza A/Sydney/2007 virus as the test antigen, 10 showed clinical signs. EI virus appeared to be readily transmitted to dogs that were held in close proximity to infected horses, but there was no evidence of lateral transmission of the virus to other dogs that did not have contact with or were not held in close proximity to horses.

Application of real-time PCR and ELISA assays for equine influenza virus to determine the duration of viral RNA shedding and onset of antibody response in naturally infected horses.

During the equine influenza (EI) outbreak, two assays were used in parallel to diagnose the disease, to demonstrate freedom from infection in disease control zones and ultimately to demonstrate that EI virus had been eliminated from the Australian horse population. A longitudinal study of a population of naturally infected horses was established to determine the performance characteristics of these assays.

An epizootic of bovine ephemeral fever in New South Wales in 2008 associated with long-distance dispersal of vectors.

OBJECTIVE: To report the rapid transmission of bovine ephemeral fever (BEF) virus from north-western New South Wales south to the Victorian border in January 2008 and to present data that suggests an uncommon meteorological event caused this rapid southward dispersal of vectors. PROCEDURE: The locations of reported clinical cases, data from sentinel herds and results from a survey of cattle in the southern affected area were examined to delineate the distribution of virus transmission. Synoptic weather charts for January 2008 were examined for meteorological conditions that may have favoured movement of vectors in a southerly direction. RESULTS: Cases of BEF and exposure to BEF virus in NSW were confirmed west of the Great Dividing Range, extending from the Queensland border to Finley, on the far North Coast and around the Hunter Valley. A low-pressure system moved south across the state on 18-19 January 2008, preceding the first cases of BEF in the south of NSW by 1-2 days. CONCLUSION: Heavy rainfall in December 2007 provided a suitable environment for vector breeding, resulting in the initiation of and support for continuing BEF virus transmission in north-western NSW. The movement of a low-pressure system south across central western NSW in mid-January 2008 after the commencement of BEF virus transmission in the north-west of the state provided a vehicle for rapid southward movement of infected vectors.

Survey of porcine circovirus 2 and postweaning multisystemic wasting syndrome in New South Wales piggeries.

OBJECTIVE: To determine if postweaning multisystemic wasting syndrome (PMWS) is occurring in the New South Wales pig population and to determine the current and past seroprevalence of porcine circovirus 2 (PCV2). DESIGN: Pig veterinarians were contacted seeking submission of tissues from animals with clinical signs suggestive of PMWS. Samples were also accepted from suspected cases of porcine dermatitis and nephropathy syndrome (PDNS). Serological studies were also undertaken on archival sera and sera submitted during the study. PROCEDURE: Histopathological examination was undertaken on all tissues submitted. The presence of PCV2 was determined by immunohistochemistry. Sera were tested for PCV2 using a commercial enzyme linked immunosorbent assay kit modified for testing of serum samples. RESULTS: No cases of PMWS were identified during the study. Four cases of PDNS were identified. PCV2 antibody was detected in 80% of archival sera from 1995 and 75.8% from 2001. Seroprevalence in samples tested during 2002-2003 was 87.8%. PCV2 was isolated from tissues of a case of PDNS. CONCLUSION: PCV2 is widespread in the New South Wales pig population and has been since at least 1995. This study describes the first isolation of an Australian PCV2. No cases of PMWS were identified in New South Wales.

Identification of a novel virus in pigs--Bungowannah virus: a possible new species of pestivirus.

In 2003 an outbreak of sudden deaths occurred in 3-4-week-old piglets on a farm in New South Wales, Australia. There was a marked increase in the birth of stillborn foetuses. Pathological changes consisted of a multifocal non-suppurative myocarditis. A viral infection was suspected but a wide range of known agents were excluded. A modified sequence independent single primer amplification (SISPA) method was used to identify a novel virus associated with this outbreak. Conserved 5'UTR motifs, the presence of a putative N(pro) coding region and limited antigenic cross-reactivity with other members of the Pestivirus genus, support the placement of this virus in the Pestivirus genus. Phylogenetic analysis of the 5'UTR, N(pro) and E2 coding regions showed this virus to be the most divergent pestivirus identified to date.

Cryptococcosis in ferrets: a diverse spectrum of clinical disease.

Cryptococcosis was diagnosed in seven ferrets (five from Australia; two from western Canada) displaying a wide range of clinical signs. Two of the ferrets lived together. One (5-years-old) had cryptococcal rhinitis and presented when the infection spread to the nasal bridge. Its sibling developed cryptococcal abscessation of the right retropharyngeal lymph node 12 months later, soon after developing a severe skin condition. DNA fingerprinting and microsatellite analysis demonstrated that the two strains isolated from these siblings were indistinguishable. Two ferrets (2- to 3-years-old) developed generalised cryptococcosis: one had primary lower respiratory tract disease with pneumonia, pleurisy and mediastinal lymph node involvement, while in the other a segment of intestine was the primary focus of infection with subsequent spread to mesenteric lymph nodes, liver and lung. The remaining three ferrets (1.75 to 4-years-old) had localised disease of a distal limb, in one case with spread to the regional lymph node. Cryptococcus bacillisporus (formerly C. neoformans var gattii) accounted for three of the four infections in Australian ferrets where the biotype could be determined. The Australian ferret with intestinal involvement and the two ferrets from Vancouver had C. neoformans var grubii infections.

Faecal viruses of dogs--an electron microscope study.

Faecal samples from 112 dogs both with and without diarrhoea were screened for parvovirus by a haemagglutination titration test and then examined by electron microscopy for the presence of viruses and virus-like particles. On the basis of morphology eight distinct viruses or virus-like particles were identified. Particles identified were coronaviruses, coronavirus-like particles, rotavirus-like particles, papovavirus-like particles, torovirus-like particles, picornavirus-like particles, 27 nm virus-like particles with projections and parvovirus-like particles which did not cause haemagglutination.