Demography of an ice-obligate mysticete in a region of rapid environmental change.

Antarctic minke whales (Balaenoptera bonaerensis, AMW) are an abundant, ice-dependent species susceptible to rapid climatic changes occurring in parts of the Antarctic. Here, we used remote biopsy samples and estimates of length derived from unoccupied aircraft system (UAS) to characterize for the first time the sex ratio, maturity, and pregnancy rates of AMWs around the Western Antarctic Peninsula (WAP). DNA profiling of 82 biopsy samples (2013-2020) identified 29 individual males and 40 individual females. Blubber progesterone levels indicated 59% of all sampled females were pregnant, irrespective of maturity. When corrected for sexual maturity, the median pregnancy rate was 92.3%, indicating that most mature females become pregnant each year. We measured 68 individuals by UAS (mean = 8.04 m) and estimated that 66.5% of females were mature. This study provides the first data on the demography of AMWs along the WAP and represents the first use of non-lethal approaches to studying this species. Furthermore, these results provide baselines against which future changes in population status can be assessed in this rapidly changing marine ecosystem.

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.

A strain-specific multiplex RT-PCR for Australian rabbit haemorrhagic disease viruses uncovers a new recombinant virus variant in rabbits and hares.

Rabbit haemorrhagic disease virus (RHDV, or GI.1) is a calicivirus in the genus Lagovirus that has been widely utilized in Australia as a biological control agent for the management of overabundant wild European rabbit (Oryctolagus cuniculus) populations since 1996. Recently, two exotic incursions of pathogenic lagoviruses have been reported in Australia; GI.1a-Aus, previously called RHDVa-Aus, is a GI.1a virus detected in January 2014, and the novel lagovirus GI.2 (previously known as RHDV2). Furthermore, an additional GI.1a strain, GI.1a-K5 (also known as 08Q712), was released nationwide in March 2017 as a supplementary tool for wild rabbit management. To discriminate between these lagoviruses, a highly sensitive strain-specific multiplex RT-PCR assay was developed, which allows fast, cost-effective and sensitive detection of the four pathogenic lagoviruses currently known to be circulating in Australia. In addition, we developed a universal RT-qPCR assay to be used in conjunction with the multiplex assay that broadly detects all four viruses and facilitates quantification of viral RNA load in samples. These assays enable rapid detection, identification and quantification of pathogenic lagoviruses in the Australian context. Using these assays, a novel recombinant lagovirus was detected in rabbit tissue samples, which contained the non-structural genes of GI.1a-Aus and the structural genes of GI.2. This variant was also recovered from the liver of a European brown hare (Lepus europaeus). The impact of this novel recombinant on Australian wild lagomorph populations and its competitiveness in relation to circulating field strains, particularly GI.2, requires further studies.

Efficacy of a commercial vaccine against different strains of rabbit haemorrhagic disease virus.

BACKGROUND: This study investigated the ability of a commercial rabbit haemorrhagic disease virus (RHDV) vaccine (Cylap®) to protect rabbits from disease caused by two different strains of the virus (v351 and K5) that are used or proposed to be used for wild rabbit control in Australia. These strains of the RHDV1 genotype belong to the 'classical RHDV' and 'antigenic variant RHDVa' subtypes, respectively. METHODS: Vaccinated rabbits were exposed to very high doses of the virus either by direct oral dosing or by exposure to infected rabbit livers. RESULTS & CONCLUSION: All vaccinated rabbits were protected against rabbit haemorrhagic disease, indicating that the Cylap® vaccine is effective against both strains of the virus under experimental conditions.

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.

Feeding rates and under-ice foraging strategies of the smallest lunge filter feeder, the Antarctic minke whale (Balaenoptera bonaerensis).

Body size and feeding mode are two fundamental characteristics that determine foraging performance and ecological niche. As the smallest obligate lunge filter feeders, minke whales represent an ideal system for studying the physical and energetic limits of filter feeding in endotherms. We used multi-sensor suction cup tags to quantify the feeding performance of Antarctic minke whales. Foraging dives around and beneath sea ice contained up to 24 lunges per dive, the highest feeding rates for any lunge-feeding whale. Their small size allows minke whales access to krill in sea-ice environments not easily accessible to larger baleen whales. Furthermore, their ability to filter feed provides an advantage over other smaller sympatric krill predators such as penguins and seals that feed on individual prey. The unique combination of body size, feeding mechanism and sea-ice habitat of Antarctic minke whales defines a previously undocumented energetic niche that is unique among aquatic vertebrates.

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.

Coronavirus infection in intensively managed cattle with respiratory disease.

BACKGROUND: A detailed laboratory investigation identified bovine coronavirus (BCoV) as the aetiological agent in an outbreak of respiratory disease at a semi-intensive beef cattle feedlot in south-east Australia. The outbreak caused 30% morbidity in the resident population and also affected two cohorts of cattle that were newly introduced to the property. METHODS: At slaughter, pulmonary consolidation and inflammatory lesions in the trachea were identified in 15 of 49 animals. Pasteurella multocida or Histophilus somni was cultured from 3 of 7 animals with lesions. Histopathological examination revealed multifocal non-suppurative bronchointerstitial pneumonia with formation of epithelial syncytial cells, sometimes associated with suppurative bronchopneumonia. RESULTS: BCoV was detected in nasal swabs and pulmonary lesions using real-time reverse transcriptase-polymerase chain reaction (qRT-PCR) assay and virus isolation. There was serological evidence of previous exposure to bovine viral diarrhoea virus, bovine respiratory syncytial virus and bovine parainfluenza virus type 3, but not to bovine herpesvirus type 1. None of these viral pathogens or Mycoplasma bovis was identified by qRT-PCR. CONCLUSION: This is believed to be the first report of BCoV in association with bovine respiratory disease complex in Australia.

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.

Longitudinal study describing the clinical signs observed in horses naturally infected with equine influenza.

We describe the clinical signs and disease course during an outbreak of equine influenza (EI) in naïve horses in a police stables in Sydney, New South Wales, Australia.

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.

Spatial association and clinical development of equine influenza in horses yarded overnight at an equestrian event at Maitland prior to propagating the 2007 epidemic in Australia.

The interaction and stabling of horses at equine events may have a substantial impact on the spread of a zoonotic disease. This study aimed to investigate the spread of equine influenza (EI) at an equestrian event at the start of the Australian outbreak. Around one-third of the competing horses were stabled overnight at the event and, of these, 70% developed symptoms of EI within 7 days. The index case was never positively identified, but stabling position and disease onset provided clues to its potential identity.

Positive results in a real-time PCR for type A influenza associated with the use of an inactivated vaccine.

A real-time reverse transcription polymerase chain reaction (qRT-PCR) test for the matrix gene of type A influenza viruses was used during the 2007 Australian equine influenza (EI) outbreak in order to confirm diagnosis and, later, eradication of the virus. During the EI outbreak, horses being exported required vaccination and individual proof of freedom from EI. At the end of the outbreak, positive results were obtained from four horses destined for export, because of contamination of the samples with the vaccine. This report highlights the need for EI testing and vaccination to occur on separate days and with the collection of swabs for testing to precede vaccination.

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.

Purification of polyclonal anti-conformational antibodies for use in affinity selection from random peptide phage display libraries: a study using the hydatid vaccine EG95.

The use of polyclonal antibodies to screen random peptide phage display libraries often results in the recognition of a large number of peptides that mimic linear epitopes on various proteins. There appears to be a bias in the use of this technology toward the selection of peptides that mimic linear epitopes. In many circumstances the correct folding of a protein immunogen is required for conferring protection. The use of random peptide phage display libraries to identify peptide mimics of conformational epitopes in these cases requires a strategy for overcoming this bias. Conformational epitopes on the hydatid vaccine EG95 have been shown to result in protective immunity in sheep, whereas linear epitopes are not protective. In this paper we describe a strategy that results in the purification of polyclonal antibodies directed against conformational epitopes while eliminating antibodies directed against linear epitopes. These affinity purified antibodies were then used to select a peptide from a random peptide phage display library that has the capacity to mimic conformational epitopes on EG95. This peptide was subsequently used to affinity purify monospecific antibodies against EG95.

Isolation of antibodies specific to a single conformation-dependant antigenic determinant on the EG95 hydatid vaccine.

EG95 is a recombinant vaccine protein that elicits protection against hydatid disease in sheep. Previous studies have shown that the host-protective epitopes on EG95 depend on correct conformation and cannot be represented by simple "linear" peptides. By screening random peptide phage display libraries with polyclonal antibodies directed against conformation-dependant epitopes of EG95, we have selected a number of peptides that mimic these epitopes. The selected peptides did not show sequence homology to EG95. Antigen binding assays involving these peptides have provided evidence of at least four conformationally-dependant epitope regions on EG95. One of the selected peptides, E100, has been used to purify antibodies from anti-sera raised in sheep vaccinated with EG95. This yielded monospecific antibodies capable of recognizing recombinant EG95 in ELISA and native EG95 in Western blot assays. This antibody was demonstrated to be effective in antibody-dependant complement-mediated in vitro killing of Echinococcus granulosus oncospheres. Peptide E100 may represent the basis for a quality control assay for EG95 production, and has the potential to become a component of a synthetic peptide-based vaccine against E. granulosus.

Comparing effectiveness of experimental and implemented bycatch reduction measures: the ideal and the real.

Fishers, scientists, and resource managers have made substantial progress in reducing bycatch of sea turtles, seabirds, and marine mammals through physical modifications to fishing gear. Many bycatch-avoidance measures have been developed and tested successfully in controlled experiments, which have led to regulated implementation of modified or new fishing gear. Nevertheless, successful bycatch experiments may not translate to effective mitigation in commercial fisheries because experimental conditions are relaxed in commercial fishing operations. Such a difference between experimental results and real-world results with fishing fleets may have serious consequences for management and conservation of protected species taken as bycatch. We evaluated preimplementation experimental measures and postimplementation efficacy from primary and gray literature for three case studies: acoustic pingers that warn marine mammals of the presence of gill nets, turtle excluder devices that reduce bycatch of turtles in trawls, and various measures to reduce seabird bycatch in longlines. Three common themes to successful implementation of bycatch reduction measures are long-standing collaborations among the fishing industry, scientists, and resource managers; pre- and postimplementation monitoring; and compliance via enforcement and incentives.

Antibody responses and epitope specificities to the Taenia solium cysticercosis vaccines TSOL18 and TSOL45-1A.

Taenia solium is a cestode parasite that causes cysticercosis in humans and pigs. This study examined the antibody responses in pigs immunized with the TSOL18 and TSOL45-1A recombinant vaccines against T. solium cysticercosis. Immunization with these proteins induced specific, complement-fixing antibodies against the recombinant antigens that are believed to be associated with vaccine-induced protection against T. solium infection. Sera from immunized pigs were used to define the linear B-cell epitopes of TSOL18 and TSOL45-1A. Prominent reactivity was revealed to one linear epitope on TSOL18 and two linear epitopes on TSOL45-1A. These, and oncosphere antigens from other taeniid cestodes, contain a protein sequence motif suggesting that they may show a tertiary structure similar to the fibronectin type III domain (FnIII). Comparison of the location of linear antigenic epitopes in TSOL18 and TSOL45-1A within the proposed FnIII structure to those within related cestode vaccine antigens reveals conservation in the positioning of the epitopes between oncosphere antigens from different taeniid species.

Molecular and genetic characterisation of the host-protective oncosphere antigens of taeniid cestode parasites.

Highly effective recombinant vaccines have been developed against Taenia ovis infection in sheep, Taenia saginata infection in cattle, Taenia solium infection in pigs, Echinococcus granulosus and Echinococcus multilocularis infections in a variety of intermediate host species. These vaccines have been based on the identification and expression in Escherichia coli of antigens derived from the oncosphere life cycle stage, contained within the parasites' eggs. Investigation of the molecular aspects of these proteins and the genes encoding them have revealed a number of common features, including the presence of a predicted secretory signal sequence, and one or two copies of a fibronectin type III domain, each encoded by separate exons within the associated gene. Evidence has been obtained to confirm glycosylation of some of these antigens. Ongoing investigations will shed light on the biological roles played by the proteins within the parasites and the mechanism by which they make the parasites vulnerable to vaccine-induced immune responses.