Evaluation of ELISA coupled with Western blot as a surveillance tool for Trichinella infection in wild boar (Sus scrofa)

Trichinella surveillance in wildlife relies on muscle digestion of large samples which are logistically difficult to store and transport in remote and tropical regions as well as labour-intensive to process. Serological methods such as enzyme-linked immunosorbent assays (ELISAs) offer rapid, cost-effective alternatives for surveillance but should be paired with additional tests because of the high false-positive rates encountered in wildlife. We investigated the utility of ELISAs coupled with Western blot (WB) in providing evidence of Trichinella exposure or infection in wild boar. Serum samples were collected from 673 wild boar from a high- and low-risk region for Trichinella introduction within mainland Australia, which is considered Trichinella-free. Sera were examined using both an 'in-house' and a commercially available indirect-ELISA that used excretory secretory (E/S) antigens. Cut-off values for positive results were determined using sera from the low-risk population. All wild boar from the high-risk region (352) and 139/321 (43.3%) of the wild boar from the low-risk region were tested by artificial digestion. Testing by Western blot using E/S antigens, and a Trichinella-specific real-time PCR was also carried out on all ELISA-positive samples. The two ELISAs correctly classified all positive controls as well as one naturally infected wild boar from Gabba Island in the Torres Strait. In both the high- and low-risk populations, the ELISA results showed substantial agreement (k-value = 0.66) that increased to very good (k-value = 0.82) when WB-positive only samples were compared. The results of testing sera collected from the Australian mainland showed the Trichinella seroprevalence was 3.5% (95% C.I. 0.0-8.0) and 2.3% (95% C.I. 0.0-5.6) using the in-house and commercial ELISA coupled with WB respectively. These estimates were significantly higher (P < 0.05) than the artificial digestion estimate of 0.0% (95% C.I. 0.0-1.1). Real-time PCR testing of muscle from seropositive animals did not detect Trichinella DNA in any mainland animals, but did reveal the presence of a second larvae-positive wild boar on Gabba Island, supporting its utility as an alternative, highly sensitive method in muscle examination. The serology results suggest Australian wildlife may have been exposed to Trichinella parasites. However, because of the possibility of non-specific reactions with other parasitic infections, more work using well-defined cohorts of positive and negative samples is required. Even if the specificity of the ELISAs is proven to be low, their ability to correctly classify the small number of true positive sera in this study indicates utility in screening wild boar populations for reactive sera which can be followed up with additional testing. (C) 2013 Elsevier B.V. All rights reserved.

Biological control of western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), in gerberas, chrysanthemums and roses

Frankliniella occidentalis (Pergande), western flower thrips (WFT), is a major worldwide pest of vegetables and ornamental crops. The biology of WFT was examined on gerberas, chrysanthemums and roses in relation to plant stage (flowering and non-flowering), pupation site, soil moisture and plant parts often inhabited by adult and immature thrips. Four foliage thrips predators ( Transeius montdorensis (Schicha), Orius armatus (Gross), Mallada signata (Schneider) and Neoseiulus cucumeris (Oudemans)) and three soil predators ( Geolaelaps aculeifer (Canestrini), Steinernema feltiae (Filipjev) and Dalotia coriaria (Kraatz)) were studied to determine their ability to reduce the numbers of WFT on gerberas, chrysanthemums and roses. There was no difference in the number of adults that emerged from growing media of high or low moisture content on any host plant. There were also no differences in the total numbers of WFT recaptured from flowering gerberas, chrysanthemums or roses. However, about seven times the number of thrips were collected from flowering chrysanthemums compared with non-flowering chrysanthemums, indicating that the flowering plants were more suitable hosts. Of all thrips recollected, the greatest percentage was immature (larval and pupal) thrips (70%, 71% and 43%) on the flowers for gerberas, chrysanthemums and roses, respectively. The mean percentage of thrips that emerged as adults from the soil was very low (5.31.2, 8.52.9, 20.59.1 and 28.25.6%) on gerberas, flowering and non-flowering chrysanthemums, and roses, respectively. Simultaneous release of foliage and soil predators did not reduce the number of thrips beyond that caused by foliage predators alone. Of the foliage predators, T. montdorensis, O. armatus and N. cucumeris performed best, significantly reducing the numbers of adult and immature thrips on flowers and foliage by 30-99%. Further research is required to determine the most cost-effective rates of release in cut flower crops.

Temporal and spatial patterns of soil water extraction and drought resistance among genotypes of a perennial C-4 grass

The objective of this study was to investigate patterns of soil water extraction and drought resistance among genotypes of bermudagrass (Cynodon spp.) a perennial C-4 grass. Four wild Australian ecotypes (1-1, 25a1, 40-1, and 81-1) and four cultivars (CT2, Grand Prix, Legend, and Wintergreen) were examined in field experiments with rainfall excluded to monitor soil water extraction at 30-190 cm depths. In the study we defined drought resistance as the ability to maintain green canopy cover under drought. The most drought resistant genotypes (40-1 and 25a1) maintained more green cover (55-85% vs 5-10%) during water deficit and extracted more soil water (120-160 mm vs 77-107 mm) than drought sensitive genotypes, especially at depths from 50 to 110 cm, though all genotypes extracted water to 190 cm. The maintenance of green cover and higher soil water extraction were associated with higher stomatal conductance, photosynthetic rate and relative water content. For all genotypes, the pattern of water use as a percentage of total water use was similar across depth and time We propose the observed genetic variation was related to different root characteristics (root length density, hydraulic conductivity, root activity) although shoot sensitivity to drying soil cannot be ruled out.

Applications of a sugar-based surveillance system to track arboviruses in wild mosquito populations

Effective arbovirus surveillance is essential to ensure the implementation of control strategies, such as mosquito suppression, vaccination, or dissemination of public warnings. Traditional strategies employed for arbovirus surveillance, such as detection of virus or virus-specific antibodies in sentinel animals, or detection of virus in hematophagous arthropods, have limitations as an early-warning system. A system was recently developed that involves collecting mosquitoes in CO2-baited traps, where the insects expectorate virus on sugar-baited nucleic acid preservation cards. The cards are then submitted for virus detection using molecular assays. We report the application of this system for detecting flaviviruses and alphaviruses in wild mosquito populations in northern Australia. This study was the first to employ nonpowered passive box traps (PBTs) that were designed to house cards baited with honey as the sugar source. Overall, 20/144 (13.9%) of PBTs from different weeks contained at least one virus-positive card. West Nile virus Kunjin subtype (WNVKUN), Ross River virus (RRV), and Barmah Forest virus (BFV) were detected, being identified in 13/20, 5/20, and 2/20 of positive PBTs, respectively. Importantly, sentinel chickens deployed to detect flavivirus activity did not seroconvert at two Northern Territory sites where four PBTs yielded WNVKUN. Sufficient WNVKUN and RRV RNA was expectorated onto some of the honey-soaked cards to provide a template for gene sequencing, enhancing the utility of the sugar-bait surveillance system for investigating the ecology, emergence, and movement of arboviruses. © 2014, Mary Ann Liebert, Inc.

Suspected Pyrrolizidine Alkaloid Hepatotoxicosis in Wild Southern Hairy-Nosed Wombats (Lasiorhinus latifrons)

Southern hairy-nosed wombats (Lasiorhinus latifrons) inhabiting degraded habitat in South Australia were recently identified with extensive hair loss and dermatitis and were in thin to emaciated body condition. Pathological and clinicopathological investigations on affected juvenile wombats identified a toxic hepatopathy suggestive of plants containing pyrrolizidine alkaloids, accompanied by photosensitive dermatitis. Hepatic disease was suspected in additional wombats on the basis of serum biochemical analysis. Preliminary toxicological analysis performed on scats and gastrointestinal contents from wombats found in this degraded habitat identified a number of toxic pyrrolizidine alkaloids consistent with ingestion of Heliotropeum europaeum. Although unpalatable, ingestion may occur by young animals due to decreased availability of preferred forages in degraded habitats and the emergence of weeds around the time of weaning of naive animals. Habitat degradation leading to malnutrition and ingestion of toxic weed species is a significant welfare issue in this species.

Increased virulence of rabbit haemorrhagic disease virus associated with genetic resistance in wild Australian rabbits (Oryctolagus cuniculus)

The release of myxoma virus (MYXV) and Rabbit Haemorrhagic Disease Virus (RHDV) in Australia with the aim of controlling overabundant rabbits has provided a unique opportunity to study the initial spread and establishment of emerging pathogens, as well as their co-evolution with their mammalian hosts. In contrast to MYXV, which attenuated shortly after its introduction, rapid attenuation of RHDV has not been observed. By studying the change in virulence of recent field isolates at a single field site we show, for the first time, that RHDV virulence has increased through time, likely because of selection to overcome developing genetic resistance in Australian wild rabbits. High virulence also appears to be favoured as rabbit carcasses, rather than diseased animals, are the likely source of mechanical insect transmission. These findings not only help elucidate the co-evolutionary interaction between rabbits and RHDV, but reveal some of the key factors shaping virulence evolution.

Demographic and functional responses of wild dogs to poison baiting

Lethal control of wild dogs - that is Dingo (Canis lupus dingo) and Dingo/Dog (Canis lupus familiaris) hybrids - to reduce livestock predation in Australian rangelands is claimed to cause continental-scale impacts on biodiversity. Although top predator populations may recover numerically after baiting, they are predicted to be functionally different and incapable of fulfilling critical ecological roles. This study reports the impact of baiting programmes on wild dog abundance, age structures and the prey of wild dogs during large-scale manipulative experiments. Wild dog relative abundance almost always decreased after baiting, but reductions were variable and short-lived unless the prior baiting programme was particularly effective or there were follow-up baiting programmes within a few months. However, age structures of wild dogs in baited and nil-treatment areas were demonstrably different, and prey populations did diverge relative to nil-treatment areas. Re-analysed observations of wild dogs preying on kangaroos from a separate study show that successful chases that result in attacks of kangaroos by wild dogs occurred when mean wild dog ages were higher and mean group size was larger. It is likely that the impact of lethal control on wild dog numbers, group sizes and age structures compromise their ability to handle large difficult-to-catch prey. Under certain circumstances, these changes sometimes lead to increased calf loss (Bos indicus/B. taurus genotypes) and kangaroo numbers. Rangeland beef producers could consider controlling wild dogs in high-risk periods when predation is more likely and avoid baiting at other times.

Haematology and Plasma Biochemistry of Wild Black Flying-Foxes, (Pteropus alecto) in Queensland, Australia

This paper establishes reference ranges for hematologic and plasma biochemistry values in wild Black flying-foxes (Pteropus alecto) captured in South East Queensland, Australia. Values were found to be consistent with those of other Pteropus species. Four hundred and forty-seven animals were sampled over 12 months and significant differences were found between age, sex, reproductive and body condition cohorts in the sample population. Mean values for each cohort fell within the determined normal adult reference range, with the exception of elevated levels of alkaline phosphatase in juvenile animals. Hematologic and biochemistry parameters of injured animals showed little or no deviation from the normal reference values for minor injuries, while two animals with more severe injury or abscessation showed leucocytosis, anaemia, thrombocytosis, hyperglobulinemia and hypoalbuminemia.

Investigation of an emerging bacterial disease in wild Queensland groper, marine fish and stingrays with production of diagnostic tools to reduce the spread of disease to other states of Australia : final report

This project describes how Streptococcus agalactiae can be transmitted experimentally in Queensland grouper. The implications of this research furthers the relatedness between Australian S. agalactiae strains from animals and humans. Additionally, this research has developed diagnostic tools for Australian State Veterinary Laboratories and Universities, which will assist in State and National aquatic animal disease detection, surveillance, disease monitoring and reporting

Diet of dingoes and other wild dogs in peri-urban areas of north-eastern Australia

Knowledge of the resource requirements of urban predators can improve our understanding of their ecology and assist town planners and wildlife management agencies in developing management approaches that alleviate human-wildlife conflicts. Here we examine food and dietary items identified in scats of dingoes in peri-urban areas of north-eastern Australia to better understand their resource requirements and the potential for dingoes to threaten locally fragmented populations of native fauna. Our primary aim was to determine what peri-urban dingoes eat, and whether or not this differs between regions. We identified over 40 different food items in dingo scats, almost all of which were mammals. Individual species commonly observed in dingo scats included agile wallabies, northern brown bandicoots and swamp wallabies. Birds were relatively common in some areas but not others, as were invertebrates. Dingoes were identified as a significant potential threat to fragmented populations of koalas. Dietary overlap was typically very high or near-identical between regions, indicating that peri-urban dingoes ate the same types or sizes of prey in different areas. Future studies should seek to quantify actual and perceived impacts of, and human attitudes towards, peri-urban dingoes, and to develop management strategies with a greater chance of reducing human-wildlife conflicts.