Presence and incidence of food-borne pathogens in Australian chicken litter.

1. Litter samples were collected at the end of the production cycle from spread litter in a single shed from each of 28 farms distributed across the three Eastern seaboard States of Australia. 2. The geometric mean for Salmonella was 44 Most Probable Number (MPN)/g for the 20 positive samples. Five samples were between 100 and 1000 MPN/g and one at 105 MPN/g, indicating a range of factors are contributing to these varying loads of this organism in litter. 3. The geometric mean for Campylobacter was 30 MPN/g for the 10 positive samples, with 7 of these samples being 100 MPN/g. The low prevalence and incidence of Campylobacter were possibly due to the rapid die-off of this organism. 4. E. coli values were markedly higher than the two key pathogens (geometric mean 20 x 105 colony forming units (cfu)/g) with overall values being more or less within the same range across all samples in the trial, suggesting a uniform contribution pattern of these organisms in litter. 5. Listeria monocytogenes was absent in all samples and this organism appears not to be an issue in litter. 6. The dominant (70% of the isolates) Salmonella serovar was S. Sofia (a common serovar isolated from chickens in Australia) and was isolated across all regions. Other major serovars were S. Virchow and S. Chester (at 10%) and S. Bovismorbificans and S. Infantis (at 8%) with these serovars demonstrating a spatial distribution across the major regions tested. 7. There is potential to re-use litter in the environment depending on end use and the support of relevant application practices and guidelines.

Characterization of the antibody response in birds following infection with wild-type and attenuated strains of Eimeria tenella and Eimeria necatrix.

Live vaccines containing attenuated parasite strains are increasingly used to control chicken coccidiosis. In this paper antibody responses elicited by infections with wild-type and attenuated strains of Eimeria tenella and E.necatrix were characterized by immunoblotting and ELISA with homologous and heterologous antisera. Few differences between antisera from birds infected with wild and attenuated strains of E. tenella were evident in immunoblots conducted with merozoite antigen preparations from both E. tenella strains, however the reactivity of sera raised in birds infected with the wild-type strain was noticeably more intense. In ELISAs conducted with merozoite antigen preparations, antisera from birds infected with the wild-type strains of E. tenella and E. necatrix consistently produced a significantly higher (P < 0.05) antibody response than antisera from birds infected with the attenuated strains. Likewise, avidity ELISAs conducted with the E. tenella strains demonstrated that antibodies in birds infected with the wild-type strain were of significantly higher avidity (P < 0.05) than antibodies in birds infected with the attenuated strain. The differences in the antibody responses are probably due to changes in the attenuated strain as a result of selection for precocious development and the less severe tissue damage and inflammation of the intestine resulting from infection with the attenuated strain.

Rapid typing of P multocida

Fowl cholera, caused by P. multocida, is a serious disease of poultry with sudden surges in mortality and an emerging disease of the free ranged poultry industries. This project will develop a more rapid and cost effective screening method for P. multocida. The impacts of this new method are manifold: It will lead to an improved understanding of the epidemiology of fowl cholera and the possible sources of entry onto the farm leading to improved biosecurity measures and control programs. Another impact is improved serotyping, which will ensure more effective and targeted vaccination programs. Improving prevention and control programs and decreasing the reliance on antibiotics will enhance the sustainability and profitability of the industry.

Characterising population structure and diversity of Australian Eimeria

Develop microsatellite markers to distinguish strains of Eimeria acervulina, E. brunetti and E. maxima. Conduct nationwide sampling of chicken faeces to build baseline of Eimeria population genetic diversity for 5 economically important speces (3 species above plus E. tenella and E. necatrix). Conduct focused local screening to assess temporal changes in populations historically sampled.

Optimise efficacy of broiler house treatments for the management of lesser mealworm

The Australian chicken meat industry requires effective agents for the management of lesser mealworm in broiler houses. The only two appropriate insecticides currently registered are cyfluthrin and spinosad, with gamma cyhalothrin being developed for registration. The industry requires the efficacy of cyfluthrin to be investigated, with progress and adoption of the latter two chemicals. Optimising the efficacy of each chemical and studying them singly and in rotation will, in addition to improving their efficacy, reduce overall insecticide use and improve their cost effectiveness.

A Differential typing of Campylobacter

Establish a DNA-based typing scheme that allows the allocation of strains of Campylobacter to types that are host specific and/or non-host specific.

Unusual growth variants of Avibacterium paragallinarum.

Two isolates of haemophilic bacteria originally isolated in the 1980s from chickens were re-examined. The addition of a 10% sterile filtrate from an overnight culture of Staphylococcus epidermidis allowed growth of both isolates in solid and liquid media that were otherwise not capable of supporting the growth of these isolates. Using the modified media, genotypic and serotypic studies were performed, which confirmed both isolates to be Avibacterium paragallinarum, with one isolate being serovar A and the other serovar C. The unusual growth requirements of these two isolates reinforces the need for careful interpretation by diagnostic laboratories examining chickens showing signs of upper respiratory tract disease.

Multidrug-resistant extraintestinal pathogenic Escherichia coli of sequence type ST131 in animals and foods.

Multidrug-resistant Escherichia colt sequence type 131 (51131) has recently emerged as a globally distributed cause of extraintestinal infections in humans. Diverse factors have been investigated as explanations for ST131's rapid and successful dissemination, including transmission through animal contact and consumption of food, as suggested by the detection of ST131 in a number of nonhuman species. For example, ST131 has recently been identified as a cause of clinical infection in companion animals and poultry, and both host groups have been confirmed as faecal carriers of ST131. Moreover, a high degree of similarity has been shown among certain ST131 isolates from humans, companion animals, and poultry based on resistance characteristics and genomic background and human and companion animal ST131 isolates tend to exhibit similar virulence genotypes. However, most ST131 isolates from poultry appear to possess specific virulence genes that are typically absent from human and companion animal isolates, including genes associated with avian pathogenic E. coli. Since the number of reported animal and food-associated ST131 isolates is quite small, the role of nonhuman host species in the emergence, dissemination, and transmission of ST131 to humans remains unclear. Nevertheless, given the profound public health importance of the emergent ST131 clonal group, even the limited available evidence indicates a pressing need for further careful study of this significant question.

The meleagrid herpesvirus 1 genome is partially resistant to transposition

The propagation of herpesvirus genomes as infectious bacterial artificial chromosomes (iBAC) has enabled the application of highly efficient strategies to investigate gene function across the genome. One of these strategies, transposition, has been used successfully on a number of herpesvirus iBACs to generate libraries of gene disruption mutants. Gene deletion studies aimed at determining the dispensable gene repertoire of the Meleagrid herpesvirus 1 (MeHV-1) genome to enhance the utility of this virus as a vaccine vector have been conducted in this report. A MeHV-1 iBAC was used in combination with the Tn5 and MuA transposition systems in an attempt to generate MeHV-1 gene interruption libraries. However, these studies demonstrated that Tn5 transposition events into the MeHV-1 genome occurred at unexpectedly low frequencies. Furthermore, characterization of genomic locations of the rare Tn5 transposon insertion events indicated a nonrandom distribution within the viral genome, with seven of the 24 insertions occurring within the gene encoding infected cell protein 4. Although insertion events with the MuA system occurred at higher frequency compared with the Tn5 system, fewer insertion events were generated than has previously been reported with this system. The characterization and distribution of these MeHV-1 iBAC transposed mutants is discussed at both the nucleotide and genomic level, and the properties of the MeHV-1 genome that could influence transposition frequency are discussed. © American Association of Avian Pathologists.

Agricultural intensification, priming for persistence and the emergence of Nipah virus: a lethal bat-borne zoonosis

Emerging zoonoses threaten global health, yet the processes by which they emerge are complex and poorly understood. Nipah virus (NiV) is an important threat owing to its broad host and geographical range, high case fatality, potential for human-to-human transmission and lack of effective prevention or therapies. Here, we investigate the origin of the first identified outbreak of NiV encephalitis in Malaysia and Singapore. We analyse data on livestock production from the index site (a commercial pig farm in Malaysia) prior to and during the outbreak, on Malaysian agricultural production, and from surveys of NiV's wildlife reservoir (flying foxes). Our analyses suggest that repeated introduction of NiV from wildlife changed infection dynamics in pigs. Initial viral introduction produced an explosive epizootic that drove itself to extinction but primed the population for enzootic persistence upon reintroduction of the virus. The resultant within-farm persistence permitted regional spread and increased the number of human infections. This study refutes an earlier hypothesis that anomalous El Nino Southern Oscillation-related climatic conditions drove emergence and suggests that priming for persistence drove the emergence of a novel zoonotic pathogen. Thus, we provide empirical evidence for a causative mechanism previously proposed as a precursor to widespread infection with H5N1 avian influenza and other emerging pathogens.

A simple, one-tube assay for the simultaneous detection and diagnosis of ten Australian poultry Eimeria

Coccidiosis is a costly worldwide enteric disease of chickens caused by parasites of the genus Eimeria. At present, there are seven described species that occur globally and a further three undescribed, operational taxonomic units (OTUs X, Y, and Z) that are known to infect chickens from Australia. Species of Eimeria have both overlapping morphology and pathology and frequently occur as mixed-species infections. This makes definitive diagnosis with currently available tests difficult and, to date, there is no test for the detection of the three OTUs. This paper describes the development of a PCR-based assay that is capable of detecting all ten species of Eimeria, including OTUs X, Y, and Z in field samples. The assay is based on a single set of generic primers that amplifies a single diagnostic fragment from the mitochondrial genome of each species. This one-tube assay is simple, low-cost, and has the capacity to be high throughput. It will therefore be of great benefit to the poultry industry for Eimeria detection and control, and the confirmation of identity and purity of vaccine strains.

Sympatric prey responses to lethal top-predator control: predator manipulation experiments

Introduction Many prey species around the world are suffering declines due to a variety of interacting causes such as land use change, climate change, invasive species and novel disease. Recent studies on the ecological roles of top-predators have suggested that lethal top-predator control by humans (typically undertaken to protect livestock or managed game from predation) is an indirect additional cause of prey declines through trophic cascade effects. Such studies have prompted calls to prohibit lethal top-predator control with the expectation that doing so will result in widespread benefits for biodiversity at all trophic levels. However, applied experiments investigating in situ responses of prey populations to contemporary top-predator management practices are few and none have previously been conducted on the eclectic suite of native and exotic mammalian, reptilian, avian and amphibian predator and prey taxa we simultaneously assess. We conducted a series of landscape-scale, multi-year, manipulative experiments at nine sites spanning five ecosystem types across the Australian continental rangelands to investigate the responses of sympatric prey populations to contemporary poison-baiting programs intended to control top-predators (dingoes) for livestock protection. Results Prey populations were almost always in similar or greater abundances in baited areas. Short-term prey responses to baiting were seldom apparent. Longer-term prey population trends fluctuated independently of baiting for every prey species at all sites, and divergence or convergence of prey population trends occurred rarely. Top-predator population trends fluctuated independently of baiting in all cases, and never did diverge or converge. Mesopredator population trends likewise fluctuated independently of baiting in almost all cases, but did diverge or converge in a few instances. Conclusions These results demonstrate that Australian populations of prey fauna at lower trophic levels are typically unaffected by top-predator control because top-predator populations are not substantially affected by contemporary control practices, thus averting a trophic cascade. We conclude that alteration of current top-predator management practices is probably unnecessary for enhancing fauna recovery in the Australian rangelands. More generally, our results suggest that theoretical and observational studies advancing the idea that lethal control of top-predators induces trophic cascades may not be as universal as previously supposed.

Protection Conferred by Infectious Coryza Vaccines Against Emergent Avibacterium paragallinarum Serovar C-1

Infectious coryza is an upper respiratory disease of chickens caused by Avibacterium paragallinarum. Outbreaks of infectious coryza caused by Av. paragallinarum serovar C-1 isolates in coryza-vaccinated flocks in Ecuador and Mexico have been reported. In the current study, the protection conferred by four commercially available, trivalent infectious coryza vaccines in chickens challenged with a serovar C-1 isolate from an apparent coryza vaccine failure in a layer flock in Mexico was evaluated. Only one infectious coryza vaccine provided a good protection level (83%) in vaccinated chickens. These results might explain the infectious coryza outbreaks in vaccinated flocks that have been observed in the field.

Mechanisms of the impact of a weed (grader grass, Themeda quadrivalvis) on reptile assemblage structure in a tropical savannah

Invasive grasses are among the worst threats to native biodiversity, but the mechanisms causing negative effects are poorly understood. To investigate the impact of an invasive grass on reptiles, we compared the reptile assemblages that used native kangaroo grass (Themeda triandra), and black spear grass (Heteropogon contortus), to those using habitats invaded by grader grass (Themeda quadrivalvis). There were significantly more reptile species, in greater abundances, in native kangaroo and black spear grass than in invasive grader grass. To understand the sources of negative responses of reptile assemblages to the weed, we compared habitat characteristics, temperatures within grass clumps, food availability and predator abundance among these three grass habitats. Environmental temperatures in grass, invertebrate food availability, and avian predator abundances did not differ among the habitats, and there were fewer reptiles that fed on other reptiles in the invaded than in the native grass sites. Thus, native grass sites did not provide better available thermal environments within the grass, food, or opportunities for predator avoidance. We suggest that habitat structure was the critical factor driving weed avoidance by reptiles in this system, and recommend that the maintenance of heterogeneous habitat structure, including clumping native grasses, with interspersed bare ground, and leaf litter are critical to reptile biodiversity.