Detection and genetic diversity of porcine rotavirus A,B and C in eastern Australian piggeries

Rotaviruses (RV) have a high prevalence in piggeries worldwide and are one of the major pathogens causing severe diarrhoea in young pigs. RV species A, B, and C have been linked to piglet diarrhoea in Australian pig herds, but their genetic diversity has not been studied in detail. Based on sequencing of the structural viral protein 7 (VP7) RVA G genotypes G3, G4 and G5, and RVC types G1, G3, G5, and G6 have been identified in Australian piggeries in previous studies. Although occurrence of RVB was reported in Australia in 1988, no further genetic analysis has been conducted. To improve health management decisions in Australian pig herds, more information on RV prevalence and genetic diversity is needed. Here, 243 enteric samples collected from 20 pig farms within Eastern Australia were analysed for the presence of RV in different age groups using a novel PCR-based multiplex assay (Pork MultiPath™ enteric panel). RVA, RVB, and RVC were detected in 10, 14, and 14 farms, respectively. Further sequencing of VP7 in selected RV-positive samples revealed G genotypes G2, G5, G9 (RVA), G6, G8, G14, G16, G20 (RVB), and G1, G3, G5, G6 (RVC) present. RVA was only detected in young (<10 weeks old) pigs whereas RVB and RVC were also detected in older animals (>11 weeks old). Interestingly, RVB and RVC G-type occurrence differed between age groups. In conclusion, this study provides new insights on the prevalence and diversity of different RV species in pig herds of Eastern Australia whilst demonstrating the ability of the Pork MultiPath™ technology to accurately differentiate between these RV species.     

Detection of Ureaplasma diversum in the upper airways of Australian feedlot cattle

Bovine respiratory disease (BRD) exerts a major impact on the beef cattle industry nationally and worldwide, with a range of aetiological factors impacting its pathogenesis. Previous research has focussed on an increasing number of bacteria and viruses that have been shown to play a role in eliciting disease. Recently, additional agents have been emerging as potential contributors to BRD, including the opportunistic pathogen Ureaplasma diversum. To determine if U. diversum was present in Australian feedlot cattle and if that presence was linked to BRD, nasal swabs were collected from a cohort of 34 hospital pen animals and compared to 216 apparently healthy animals sampled contemporaneously at feedlot induction and again after 14 days on feed at an Australian feedlot. All samples were subjected to a de novo polymerase chain reaction (PCR) assay targeting U. diversum in combination with other BRD agents. U. diversum was detected at a low prevalence in cattle at induction (Day 0: 6.9%, Day 14: 9.7%), but in a significantly greater proportion of cattle sampled from the hospital pen (58.8%). When considering the presence of other BRD-associated agents, co-detection of U. diversum and Mycoplasma bovis was most common in hospital pen animals receiving treatment for BRD. These findings suggest that U. diversum may be an opportunistic pathogen involved in the aetiology of BRD in Australian feedlot cattle, in combination with other agents, with further studies are warranted to identify if a causal relationship exists.     

High yield and rapid growth of Neoparamoeba pemaquidensis in co-culture with a rainbow trout gill-derived cell line RTgill-W1

Neoparamoeba pemaquidensis is an ubiquitous amphizoic marine protozoan and has been implicated as the causative agent for several diseases in marine organisms, most notably amoebic gill disease (AGD) in Atlantic salmon. Despite several reports on the pathology of AGD, relatively little is known about the protozoan and its relationship to host cells. In this study, an in vitro approach using monolayers of a rainbow trout gill cell line (RTgill-W1, ATCC CRL-2523) was used to rapidly grow large numbers of N. pemaquidensis (ATCC 50172) and investigate cell-pathogen interactions. Established cell lines derived from other tissues of rainbow trout and other fish species were also evaluated for amoeba growth support. The amoebae showed preference and highest yield when grown with RTgill-W1 over nine other tested fish cell lines. Amoeba yields could reach as high as 5 x 10(5) cells mL(-1) within 3 days of growth on the gill cell monolayers. The amoebae caused visible focal lesions in RTgill-W1 monolayers within 24 h of exposure and rapidly proliferated and spread with cytopathic effects destroying the neighbouring pavement-like cells within 48-72 h after initial exposure in media above 700 mOsm kg(-1). Disruption of the integrity of the gill cell monolayers could be noted within 30 min of exposure to the amoeba suspensions by changes in transepithelial resistance (TER) compared with control cell monolayers maintained in the exposure media. This was significantly different by 2 h (P < 0.05) compared with control cells and remained significantly different (P < 0.01) for the remaining 72 h that the TER was monitored. The RTgill-W1 cell line is thus a convenient model for growing N. pemaquidensis and for studying host-pathogen interactions in AGD.     

Alpha-amylase/subtilisin Inhibitor Levels in Australian Barleys

An enzyme-linked immunosorbent assay (ELISA) was used to determine the bifunctional alpha-amylase/subtilisin inhibitor (BASI) content of barley grain from 11 cultivars grown in six diverse locations in Australia. The inhibitor ranged from 119 to 254 μg/g in 57 barley samples. Genotype had a significant (P<0·05) effect on BASI content but there was no effect due to environment. Total protein varied independently of BASI and was influenced by environment and genotype. BASI content was higher (P<0·05) in malting barley than in feed barley and was correlated positively (r=0·29;P<0·05) with alpha-amylase activity in corresponding malts. The ELISA used monoclonal and polyclonal antibodies raised against purified BASI. In immunoblot analysis the monoclonal antibody showed high specificity for the inhibitor in barley and also detected the inhibitor in wheat. Low levels of inhibitor (mean 3·2 μg/g) were found in 12 Australian wheat cultivars using the ELISA developed for barley. The assay had a linear working range of 5–50 ng/mL with a detection limit of 2 ng/mL. Reproducibility between assays was good (CV=4·9%) but mean recoveries were high, ranging from 116–129% when purified inhibitor was added to barley extracts. The ELISA may have useful applications in brewing research and barley breeding programmes.     

Evaluation of ear tags impregnated with cypermethrin for the control of sheep body lice (Damalinia ovis)

The application of polymer matrix ear tags impregnated with 8.5% w/w cypermethrin to 6 wethers following shearing reduced lice to non-detectable levels on 4 of them at 29 weeks after tagging. At the conclusion of the study at 45 weeks the mean count of lice on tagged wethers was 3 per sheep compared to 158 on untreated wethers. In a second experiment, 6 non-infected sheep were treated at shearing with cypermethrin tags, 6 with 25 g/l cypermethrin backline formulation, 6 with tags plus backline and 6 were left untreated. Each group was exposed to 6 sheep with moderate to heavy infestations of lice. Compared to controls, all treatments delayed infestation, but cypermethrin tags gave no longer protection than backline treatment. All sheep were infested by 17 weeks after commencement of the study. At 45 weeks mean counts of lice were 38, 27, 20 and 74 respectively for sheep treated with tags, backline formulation, backline plus tags and untreated. Possible reasons for the better effect observed from applying tags to infested sheep than to sheep which were not infected at application but which were subsequently exposed to infested sheep are discussed.     

Development of a forest certification standard compatible with PEFC and FSC's management requirements. A case study from Greece

Greece currently has no national forest certification standard.This paper explains how a draft forest management standard forGreece was developed to be compatible with both the Forest StewardshipCouncil and the Programme for the Endorsement of Forest Certificationschemes requirements. The draft standard was tested in two contrastingforest areas to investigate its ease of operation, to indicatenecessary refinements to the standard and to reveal major areasof weakness in current forest management practices. Field testingshowed that the standard was operationally efficient and thatrelatively few changes to the standard were necessary. Majorweaknesses in current management practices were identified asbreaches of health and safety, poor training of forest workersand inadequate consultation with stakeholders.     

New Zealand’s Cetaceans and Pinnipeds

The 8 species of baleen whale in New Zealand waters range in length from 6 to 30+ metres, and the 25 species of toothed whale range up to 18 metres. A single porpoise species is known in New Zealand sub-Antarctic waters.

The baleen whales and many of the toothed whales are migratory, and New Zealand straddles their well-defined routes: a combination which allowed whaling to flourish. The toothed whales are the more gregarious and may indulge in mass strandings. Current research aims to investigate the biology of all of the whale species.

The otariid, or eared seals found in the New Zealand region are the New Zealand sealion and the New Zealand fur seal. The commonest of the phocid, or earless seals on the New Zealand coast are the elephant and leopard seals. All of these breed in the southern parts of the region but range further north. The fur seal population is rising slowly, but the small population size of the New Zealand sealion gives cause for concern.

Marine mammals form an important part of the fauna of the New Zealand region, and veterinary knowledge and general research should be increased.