Mark O'Dea

Variation in the responses of wild species of duck, gull, and wader to inoculation with a wild-bird-origin H6N2 low pathogenicity avian influenza virus

SUMMARY There is poor understanding of host responses to avian influenza virus (AIV) infection in wild birds, with most experimental studies using captive-bred birds and highly pathogenic AIVs that have an early endpoint. The objective of this study was to experimentally assess antibody responses and patterns of viral excretion in wild birds challenged with a low pathogenicity AIV. Ruddy turnstones (Arenaria interpres), silver gulls (Chroicocephalus novaehollandiae), and wandering whistling ducks (Dendrocygna arcuata) were challenged with a H6N2 virus, and blood, cloacal, and oropharyngeal (OP) swabs were analyzed from each bird over 28 days, with serology conducted on the ducks for a further 7 mo. Nineteen of 22 birds showed evidence of infection, with respiratory infection prevalent in the turnstones and gulls as mostly low titer viral excretion to 4 days postinoculation (DPI) with gastrointestinal replication detected in only one turnstone. In AIV naive ducks, there was gastrointestinal tropism with moderately high titer viral excretion via the cloaca to 6 DPI and low-grade OP viral excretion to 4 DPI. The hemagglutination inhibition antibody response was poor in the ducks, declining from 19 to 56 DPI, with higher titer responses in the gulls and turnstones. All infected birds responded with elevated nucleoprotein antibodies (in competitive enzyme-linked immunosorbent assay) by 7–10 DPI, and in the ducks these waned slowly after 42 DPI and were long-lived to at least 8 mo. The interspecies variability in response was consistent with a subtype that had adapted well in ducks, while the response of the turnstones may have been influenced by preexisting immunity to AIV. These findings provide insight into AIV infection dynamics in wild birds and highlight the need for further research. RESUMEN Variación en las respuestas de las especies silvestres de pato, gaviota y aves limícolas a la inoculación con un virus de influenza aviar de baja patogenicidad de origen aviar subtipo H6N2. Existe poca información sobre las respuestas del huésped a la infección por el virus de la influenza aviar (AIV) en aves silvestres, con la mayoría de estudios experimentales realizados con aves criadas en cautividad y los virus de influenza aviar de alta patogenicidad que tienen un punto final temprano. El objetivo de este estudio fue evaluar experimentalmente las respuestas de anticuerpos y los patrones de excreción viral en las aves silvestres expuestas a un virus de influenza aviar de baja patogenicidad. Vuelvepiedras comunes (Arenaria interpres), gaviotas plateadas (Larus novaehollandiae) y yaguasas errantes (Dendrocygna arcuata) fueron expuestas a un virus H6N2, y muestras de sangre, hisopos cloacales y orofaríngeos se analizaron de cada ave durante 28 días, la serología se llevó a cabo en los patos por siete meses. Diecinueve de 22 aves mostraron evidencia de infección, con infección respiratoria frecuente en los vuelvepiedras y en las gaviotas principalmente con excreción viral con título bajo a los cuatro días después de la inoculación, con replicación gastrointestinal detectada en un vuelvepiedras. En patos no expuestos al virus de la influenza aviar, había tropismo gastrointestinal con una excreción con título moderadamente alto a través de la cloaca a los seis días después de la inoculación y con un grado bajo de excreción viral orofaríngea a los cuatro días después de la infección. La respuesta por anticuerpos inhibidores de la hemaglutinación era pobre en los patos, que declinó de 19 a 56 días después de la inoculación, con respuestas de títulos superiores en las gaviotas y en los vuelvepiedras. Todas las aves infectadas respondieron con anticuerpos elevados contra la nucleoproteína elevadas (ensayo competitivo de inmunoabsorción con enzimas ligadas) de siete a diez días después de la inoculación, y en los patos éstos títulos disminuyeron lentamente después de los 42 días después de la inoculación y eran de larga vida hasta por lo menos ocho meses. La variabilidad de la respuesta entre las especies fue consistente con un subtipo que se había adaptado bien en patos, mientras que la respuesta de los vuelvepiedras pudo haber sido influenciada por la inmunidad preexistente al virus de influenza aviar. Estos resultados proporcionan una idea de la dinámica de la infección por el virus de influenza aviar en aves silvestres y ponen de manifiesto la necesidad de más investigaciones.

High prevalence of Trypanosoma vegrandis in bats from Western Australia

Abstract The present study describes the first report of Trypanosoma vegrandis in bats using morphology and sequence analysis of the 18S rRNA gene. The PCR prevalence of T. vegrandis in bats was 81.8% (18/22). The high prevalence of T. vegrandis in the present study suggests that bats may play an important role in the epidemiology of T. vegrandis in Australia. T. vegrandis appears to be geographically dispersed, has a wide distribution in Australia and low levels of host specificity.

Avian influenza in Australia : a summary of 5 years of wild bird surveillance

BACKGROUND Avian influenza viruses (AIVs) are found worldwide in numerous bird species, causing significant disease in gallinaceous poultry and occasionally other species. Surveillance of wild bird reservoirs provides an opportunity to add to the understanding of the epidemiology of AIVs. METHODS This study examined key findings from the National Avian Influenza Wild Bird Surveillance Program over a 5-year period (July 2007-June 2012), the main source of information on AIVs circulating in Australia. RESULTS The overall proportion of birds that tested positive for influenza A via PCR was 1.9 ± 0.1%, with evidence of widespread exposure of Australian wild birds to most low pathogenic avian influenza (LPAI) subtypes (H1-13, H16). LPAI H5 subtypes were found to be dominant and widespread during this 5-year period. CONCLUSION Given Australias isolation, both geographically and ecologically, it is important for Australia not to assume that the epidemiology of AIV from other geographic regions applies here. Despite all previous highly pathogenic avian influenza outbreaks in Australian poultry being attributed to H7 subtypes, widespread detection of H5 subtypes in wild birds may represent an ongoing risk to the Australian poultry industry.

Porcine circovirus‐associated disease in weaner pigs in Western Australia

OBJECTIVE To report the occurrence and pathology of porcine circovirus type 2 (PCV2)-associated disease (PCVAD) of postweaning pigs in two Australian pig herds. METHODS Mortality data from two commercial piggeries that experienced higher than normal postweaning illthrift and mortalities were examined. Gross and histopathological examinations were performed on the index cases, and at weekly intervals thereafter for a period of 10 weeks. Specimens were submitted to the laboratory for routine diagnostic testing and for exclusion of porcine reproductive and respiratory syndrome virus (PRRSV). The genomes of two strains of PCV2 isolated during testing were sequenced. RESULTS Mortality rates in weaned, 5-12-week-old pigs spiked significantly during mid to late 2007. This increase in the mortalities was mainly attributed to salmonella-associated diarrhoea and illthrift. Salmonellosis was diagnosed in 73/110 cases inclusive of both piggeries. Many pigs also had chronic granulomatous lymphadenitis and diffuse histiocytic interstitial pneumonia consistent with PCVAD and associated with varying amounts of PCV2 antigen and inclusion bodies. All samples tested for PRRSV were negative. Sequence analysis of the PCV2 isolates showed strain differences between piggeries. CONCLUSION This report describes the first outbreaks of PCVAD in growing pigs in Western Australia (WA) and describes lesions not previously seen in this laboratory. It also describes the first isolation of a PCV2 group 1 virus in WA associated with PCVAD. Although the outbreaks of PCVAD occurred with concurrent salmonellosis, the two diseases were unrelated. Neither of the outbreaks met the Australian case definition for the diagnosis of postweaning multisystemic wasting syndrome.

A virological investigation into declining woylie populations

Abstract. The woylie (Bettongia penicillata ogilbyi) is a critically endangered small Australian marsupial that is in a state of accelerated population decline for reasons that are currently unknown. The aim of the present study was to elucidate the involvement of several viral pathogens through strategic serological testing of several wild woylie populations. Testing for antibodies against the Wallal and Warrego serogroup of orbiviruses, Macropod herpesvirus 1 and Encephalomyocarditis virus in woylie sera was undertaken through virus neutralisation tests. Moreover, testing for antibodies against the the alphaviruses Ross River virus and Barmah Forest virus and the flaviviruses Kunjin virus and Murray Valley encephalitis virus was undertaken through virus neutralisation tests and ELISA mainly because of the interest in the epidemiology of these important zoonoses as it was considered unlikely to be the cause of the decline. Between 15 and 86 samples were tested for each of the four sites in south-western Australia (Balban, Keninup, Warrup and Karakamia). Results indicated no exposure to any of the viral pathogens investigated, indicating that all populations are currently naïve and may be at risk if these pathogens were to be introduced.

Chemical Synthesis, Characterisation, and Biocompatibility of Nanometre Scale Porous Anodic Aluminium Oxide Membranes for Use as a Cell Culture Substrate for the Vero Cell Line: A Preliminary Study

In this preliminary study we investigate for the first time the biomedical potential of using porous anodic aluminium oxide (AAO) membranes as a cell substrate for culturing the Cercopithecus aethiops (African green monkey) Kidney (Vero) epithelial cell line. One advantage of using the inorganic AAO membrane is the presence of nanometre scale pore channels that allow the exchange of molecules and nutrients across the membrane. The size of the pore channels can be preselected by adjusting the controlling parameters of a temperature controlled two-step anodization process. The cellular interaction and response of the Vero cell line with an in-house synthesised AAO membrane, a commercially available membrane, and a glass control were assessed by investigating cell adhesion, morphology, and proliferation over a 72 h period. The number of viable cells proliferating over the respective membrane surfaces revealed that the locally produced in-house AAO membrane had cells numbers similar to the glass control. The study revealed evidence of focal adhesion sites over the surface of the nanoporous membranes and the penetration of cellular extensions into the pore structure as well. The outcome of the study has revealed that nanometre scale porous AAO membranes have the potential to become practical cell culture scaffold substrates with the capability to enhance adhesion and proliferation of Vero cells.

Prevalence, risk factors and antimicrobial resistance of Salmonella diarrhoeal infection among children in Thi-Qar Governorate, Iraq

We conducted a hospital-based cross-sectional study among children aged <5 years in Thi-Qar Governorate, south-eastern Iraq, in order to examine the prevalence, risk factors and antimicrobial resistance associated with gastroenteritis caused by Salmonella infection. From 320 diarrhoea cases enrolled between March and August 2016, 33 (10·3%, 95% confidence interval (CI) 8·4-12·4) cases were stool culture-positive for non-typhoidal Salmonella enterica. The most commonly identified serovar was Typhimurium (54%). Multivariable logistic regression analysis indicated that the odds of Salmonella infection in children from households supplied by pipe water was 4·7 (95% CI 1·6-13·9) times higher compared with those supplied with reverse osmosis treated water. Similarly, children from households with domestic animals were found to have a higher odds (OR 10·5; 95% CI 3·8-28·4) of being Salmonella stool culture-positive. The likelihood of Salmonella infection was higher (OR 3·9; 95% CI 1·0-6·4) among children belonging to caregiver with primary vs. tertiary education levels. Lower odds (OR 0·4; 95% CI 0·1-0·9) of Salmonella infection were associated with children exclusively breast fed as compared with those exclusively bottle fed. Salmonella infection was three times lower (95% CI 0·1-0·7) in children belonging to caregiver who reported always washing hands after cleaning children following defecation, vs. those belonging to caregivers who did not wash hands. The antimicrobial resistance profile by disc diffusion revealed that non-susceptibility to tetracycline (78·8%), azithromycin (66·7%) and ciprofloxacin (57·6%) were the most commonly seen, and 84·9% of Salmonella isolates were classified as multi-drug resistant. This is the first study on prevalence and antimicrobial resistance of Salmonella infection among children in this setting. This work provides specific epidemiological data which are crucial to understand and combat paediatric diarrhoea in Iraq.

Alphaherpesvirus-associated disease in greater bilbies (Macrotis lagotis)

CASE REPORT A captive breeding colony of 9 greater bilbies (Macrotis lagotis) exhibited mild upper respiratory signs and sudden deaths with 100% mortality over a 2-week period. Histologically, acute necrotising and erosive epithelial lesions throughout the upper respiratory system and bronchi were associated with eosinophilic intranuclear inclusion bodies. Inclusions were also present in hepatocytes and adrenocortical cells, but were not always associated with necrosis. Transmission electron microscopy of lung sections revealed nucleocapsids forming arrays within some nuclei. A pan-herpesvirus PCR yielded a 440-bp product, with sequencing confirming homology with the alphaherpesviruses. Viral culture in a marsupial cell line resulted in cytopathic effect consistent with an alphaherpesvirus. CONCLUSION This is the first report of a herpesvirus-associated disease in greater bilbies.

Giardia duodenalis mouse model for the development of novel antigiardial agents

This study describes a neonatal mouse model of Giardia infection for development of novel antigiardials. Mice were infected with the axenically cultured Assemblage A BAH2c2 strain, with 105 trophozoites per animal recovered. This model proved to be robust and consistent for use in preliminary drug efficacy trials and drug development.

Occurrence, antimicrobial resistance and whole-genome sequencing analysis of Salmonella isolates from chicken carcasses imported into Iraq from four different countries

Salmonella is a major cause of human foodborne illnesses worldwide; however, little is known about its occurrence and genomic characteristics in food sources in many developing countries. This study investigates the occurrence, serotypes distribution, antimicrobial resistance, and multilocus sequence types (ST) of Salmonella isolated from 400 imported frozen chicken carcasses sold in the markets of Thi-Qar, south-eastern Iraq. Salmonella was detected in 46 out of 400 tested samples [11.5% (95% confidence interval: 8.5%-15.0%)]. S. Typhimurium was the most abundant (30.4%) among 14 different serotypes recovered from the tested frozen carcasses. Antimicrobial resistance was most frequently detected against tetracycline (84.4%), nalidixic acid (80.4%), streptomycin (69.6%) and trimethoprim/sulfamethoxazole (65.2%). Whole-genome sequencing (WGS) analysis revealed that 18 isolates harbored four β-lactamase resistance genes, with blaCARB-2 was the most commonly (14/18) detected. It was possible to identify 8 multilocus sequence types from the WGS analysis of 40 out of the 46 Salmonella isolates; with ST-11 (among S. Enteritidis) and ST-19 (among S. Typhimurium) were the most frequently detected. These results add to our understanding of the global epidemiology of Salmonella. Our work stands as one of the first reports on WGS analysis of Salmonella from retail chicken in a Middle-Eastern country. Results from this study could be valuable for guiding an informed import risk analysis aiming at reducing the exposure risk from Salmonella through imported chicken carcasses into Iraq. This work demonstrates the value of WGS as a promising tool for supporting evidence-based food safety hazard characterization.