Distributions of lesser mealworm (Coleoptera: Tenebrionidae) in litter of a compacted earth floor broiler house in subtropical Queensland, Australia

Distributions of lesser mealworm, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae), in litter of a compacted earth floor broiler house in southeastern Queensland, Australia, were studied over two flocks. Larvae were the predominant stage recorded. Significantly low densities occurred in open locations and under drinker cups where chickens had complete access, whereas high densities were found under feed pans and along house edges where chicken access was restricted. For each flock, lesser mealworm numbers increased at all locations over the first 14 d, especially under feed pans and along house edges, peaking at 26 d and then declining over the final 28 d. A life stage profile per flock was devised that consisted of the following: beetles emerge from the earth floor at the beginning of each flock, and females lay eggs, producing larvae that peak in numbers at 3 wk; after a further 3 to 4 wk, larvae leave litter to pupate in the earth floor, and beetles then emerge by the end of the flock time. Removing old litter from the brooder section at the end of a flock did not greatly reduce mealworm numbers over the subsequent flock, but it seemed to prevent numbers increasing, while an increase in numbers in the grow-out section was recorded after reusing litter. Areas under feed pans and along house edges accounted for 5% of the total house area, but approximately half the estimated total number of lesser mealworms in the broiler house occurred in these locations. The results of this study will be used to determine optimal deployment of site-specific treatments for lesser mealworm control.

Development and validation of an ELISA for detecting antibodies to Eimeria tenella in chickens

The aim of this study was to develop and validate an ELISA for detecting chicken antibodies to Eimeria tenella. An initial comparison of merozoite and sporozoite antigen preparations revealed few differences in their ability to monitor the onset, kinetics and magnitude of the antibody response suggesting that both antigens would be equally useful for development of an ELISA. Furthermore the cross-reactivity of these antigens with sera from birds infected with chicken Eimeria species was similar. The merozoite antigen was selected for further evaluation because it was easier to prepare. Discrimination between sera from birds experimentally infected with E. tenella and birds maintained in an Eimeria-free isolation facility was excellent. In sera collected from free-range layers and commercial broilers there also appeared to be clear discrimination between infected and uninfected birds. The ELISA should prove useful for monitoring infectivity in vaccination programmes in layer and breeder flocks and for assessing the effectiveness of biosecurity measures in broiler flocks.

Antibody response against endogenous stages of an attenuated strain of Eimeria tenella

The application of attenuated vaccines for the prevention of chicken coccidiosis has increased exponentially in recent years. In Eimeria infections, protective immunity is thought to rely on a strong cell mediated response with antibodies supposedly playing a minor role. However, under certain conditions antibodies seem to be significant in protection. Furthermore, antibodies could be useful for monitoring natural exposure of flocks to Eimeria spp. and for monitoring the infectivity of live vaccines. Our objective was to investigate the chicken antibody response to the different parasite lifecycle stages following infection with an attenuated strain of Eimeria tenella. Western blotting analysis of parasite antigens prepared from the lining of caeca infected with the attenuated strain of E. tenella revealed two dominant antigens of 32 and 34 kDa, apparently associated with trophozoites and merozoites that were present at high concentrations between 84 and 132 h post-infection. When cryosections of caeca infected with E. tenella were probed with IgY purified from immune birds the most intense reaction was observed with the asexual stages. Western blotting analysis of proteins of purified sporozoites and third generation merozoites and absorption of stage-specific antibodies from sera suggested that a large proportion of antigens is shared by the two stages. The time-courses of the antibody response to sporozoite and merozoite antigens were similar but varied depending on the inoculation regime and the degree of oocyst recirculation.

Decoration of Pasteurella multocida lipopolysaccharide with phosphocholine is important for virulence

Phosphocholine (PCho) is an important substituent of surface structures expressed by a number of bacterial pathogens. Its role in virulence has been investigated in several species, in which it has been shown to play a role in bacterial adhesion to mucosal surfaces, in resistance to antimicrobial peptides, or in sensitivity to complement-mediated killing. The lipopolysaccharide (LPS) structure of Pasteurella multocida strain Pm70, whose genome sequence is known, has recently been determined and does not contain PCho. However, LPS structures from the closely related, virulent P. multocida strains VP161 and X-73 were shown to contain PCho on their terminal galactose sugar residues. To determine if PCho was involved in the virulence of P. multocida, we used subtractive hybridization of the VP161 genome against the Pm70 genome to identify a four-gene locus (designated pcgDABC) which we show is required for the addition of the PCho residues to LPS. The proteins predicted to be encoded by pcgABC showed identity to proteins involved in choline uptake, phosphorylation, and nucleotide sugar activation of PCho. We constructed a P. multocida VP161 pcgC mutant and demonstrated that this strain produces LPS that lacks PCho on the terminal galactose residues. This pcgC mutant displayed reduced in vivo growth in a chicken infection model and was more sensitive to the chicken antimicrobial peptide fowlicidin-1 than the wild-type P. multocida strain

Evaluation of the most effective measures of infectivity of a precocious line of Eimeria acervulina in poultry

Coccidiosis is an economically important parasitic disease of chickens that, in Australia, is caused by seven species of the genus Eimeria.1 The disease has traditionally been controlled by prophylactic drugs, but vaccination with attenuated lines of the parasites2–4 is rapidly gaining acceptance world wide. Live Eimeria vaccines are produced in batches which are not frozen and have a limited shelf life. The per cent infectivity of vaccine seed stocks and the vaccines produced from them must therefore be accurately monitored using standardised dose dependant assays to ensure that shelf life, quality control and vaccine release specifications are met. Infectivity for the chicken host cannot readily be determined by microscopic observation of oocysts or sporocyst hatching.5 Dose dependent parameters such as body weight gain, feed conversion ratio, visual lesion scores, mortality, oocysts production, clinical symptoms and microscopic lesion counts could be used as measures of infectivity.6–11 These parameters show significant dose dependant effects with field strains, but lines of vaccine parasites that have been selected for precocious development with associated reduced virulence and reproductive capability may not have the same effect.3,4 The aim of this trial was to determine which parameters provide the most effective measures of infective dose in birds inoculated with a precocious vaccine strain.

Persistence of sodium monofluoroacetate (1080) in fox baits and implications for fox management in south-eastern Australia

The persistence of 1080 in two commonly used fox baits, Foxoff ® and chicken wingettes, was assessed under different climatic and rainfall conditions in central-western New South Wales. The rate of 1080 degradation did not change significantly between the Central Tablelands and the relatively hotter and drier environment of the Western Slopes. Loss of 1080 from wingettes was independent of the rainfall and climate conditions, with wingettes remaining lethal to foxes for, on average, 0.9 weeks. Foxoff ® baits remained lethal for longer than wingettes under all tested conditions, although their rate of degradation increased generally with increasing rainfall. As a result, areas baited with Foxoff® will require longer withholding periods for working dogs than those baited with wingettes, especially during dry periods. Wingettes may have advantages for use in sensitive areas where long-term hazards from toxic baits are undesirable. We found significant variations in 1080 concentration in freshly prepared baits that may result in efficacy, non-target and regulatory concerns for baiting campaigns. As a result, the superior quality control and shelf-stability of manufactured Foxoff® may be important criteria for favouring its use over freshly prepared bait types. However, use strategies for any bait type must ensure that foxes consume lethal doses of 1080 to avoid potential problems such as the development of learned aversion to baits or pesticide resistance.

Diagnosis of a naturally occurring dual infection of layer chickens with fowlpox virus and gallid herpesvirus 1 (infectious laryngotracheitis virus).

An outbreak of acute respiratory disease in layers was diagnosed as being of dual nature due to fowlpox and infectious laryngotracheitis using a multidisciplinary approach including virus isolation, histopathology, electron microscopy and polymerase chain reaction (PCR). The diagnosis was based on virus isolation of gallid herpesvirus 1 (GaHV-1) in chicken kidney cells and fowlpox virus (FWPV) in 9-day-old chicken embryonated eggs inoculated via the chorioallantoic membrane. The histopathology of tracheas from dead birds revealed intra-cytoplasmic and intra-nuclear inclusions suggestive of poxvirus and herpesvirus involvement. The presence of FWPV was further confirmed by electron microscopy, PCR and histology. All FWPV isolates contained the long terminal repeats of reticuloendotheliosis virus as demonstrated by PCR. GaHV-1 isolates were detected by PCR and were shown to have a different restriction fragment length polymorphism pattern when compared with the chicken embryo origin SA2 vaccine strain; however, they shared the same pattern with the Intervet chicken embryo origin vaccine strain. This is a first report of dual infection of chickens with GaHV-1 and naturally occurring FWPV with reticuloendotheliosis virus insertions. Further characterization of the viruses was carried out and the results are reported here.

Factors affecting localized abundance and distribution of lesser mealworm in earth-floor broiler houses in subtropical Australia.

Factors that influence the localized abundance and distribution of lesser mealworm, Alphitobius diaperinus (Panzer), in litter of two compacted earth-floor broiler houses in subtropical Australia were studied using various experimental manipulations. Numbers of lesser mealworms substantially increased inside caged areas and under uncaged empty feed pans placed in open areas of the houses. These populations were found to be localized and independent of chicken-feed, manure, and high beetle populations that normally occur under existing feed pans. Substantial horizontal movement of larvae to under feed pans was recorded. Placing metal barriers around these pans significantly restricted this movement. In almost all treatments, lesser mealworms typically peaked in numbers during the middle of the flock time. This temporal pattern of abundance also was observed under pans within barriers, where relatively low insect numbers occurred, but it was not observed in uncaged open areas (where chickens had complete access). It is likely that larvae do not establish in open areas, but fluctuate in numbers as they either move to refuges away from chickens or suffer high rates of mortality. In these refuges, larvae peak in numbers and then leave the litter environment to pupate in the earth floor before the end of the flock time. This behavior might be exploited for management of lesser mealworm by targeting applications of control agents.

The effect of foliar sprays of the synthetic auxin 3-5-6 TPA, on fruit drop, fruit size and seed development in three lychee (Litchi chinensis) cultivars: Tai So, Fay Zee Sui and Kwai Mai Pink.

Fruit drop in lychee can cause major yield losses in Australia, the severity varying with cultivar and season. Research in China, South Africa and Israel has demonstrated the potential for synthetic auxins used as foliar sprays to reduce fruit drop in lychee. Trials were initiated in Australia to test the efficacy of the synthetic auxin, 3-5-6 Trichloro-2-phridyl-oxyacetic acid (3-5-6 TPA) at 50 ppm on the cultivars Tai So, Fay Zee Sui and Kwai Mai Pink. Results indicate that in most cases the TPA reduced natural fruit drop however the size of the fruit at the time of application affects the response and the ideal application time varies with cultivar; approximately 13 mm fruit length in 'Kwai Mai Pink', 20 mm in 'Fay Zee Sui' and 27 mm in 'Tai So'. If applied too early in 'Tai So', it caused an increase in fruit drop. The TPA was most effective when natural fruit drop was high, reducing fruit drop from 74.7 to 34.9% in 'Kwai Mai Pink' and least effective when natural fruit drop was low. An increase in the percentage of fruit with poorly developed (chicken tongue) seed and slightly larger fruit size was also observed in treated trees.

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.

Lot feeding and intensive livestock manure and effluent management

Manure management emissions may present much greater opportunity for greenhouse gas mitigation in the feedlot, pig, chicken meat, egg and diary industries, than the current IPCC and DCC calculation guidelines suggest. Current literature and understanding of manure mass loss throughout the manure management system does not support these current guidelines; in which the emission rates are fixed and consequently don't allow incentives for reduced emissions.

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.

Trialling Biological Agents for the Management of Lesser Mealworm in Australian Broiler Houses

For approximately three decades the Australian broiler industry has relied heavily on the use of insecticides as its key tool for management of darkling beetle or lesser mealworm, Alphitobius diaperinus [Panzer] in broiler houses. The use of these chemicals over this period has been largely unchecked which has resulted in the development of strong insecticide resistance in many beetle populations from broiler farms. Although we are in a period now with an improved knowledge of managing resistance and the availability of new more effective insecticides that are currently marketed, the industry still requires more pest management options in order to inhibit development of resistance and reduce overall chemical use. In response to this need, ‘natural’ agents such as entomopathogenic nematodes and fungi were proposed as potential agents for managing darkling beetle populations in Australian broiler houses. Since 2007 laboratory and field studies have been undertaken to assess these agents. This report outlines these studies and discusses potential benefits to the Chicken Meat industry resulting from this research.

Food-borne pathogens - a key issue for all food animal industries

The modern consumer has an attitude that food safety is non-negotiable issue – the consumer simply demands food to be safe. Yet, at the same time, the modern consumer has an expectation that the food safety is the responsibility of others – the primary producer, the processing company, the supermarket, commercial food handlers and so on. Given this environment, all food animal industries have little choice but to regard food safety as a key issue. As an example, the chicken meat industry, via the two main industry funding bodies – the Rural Industries Research and Development Corporation (Chicken Meat) and the Poultry CRC – has a comprehensive research program that seeks to focus on reducing the risks of food-borne diseases at all points of the food processing chain – from the farm to the processing plant. The scale of the issue for all industries can be illustrated by an analysis of the problem of campylobacterosis – a major food-borne disease. It has been estimated that there are around 230,000 cases of campylobacterosis per year. In 1995, it was estimated that each case of food-borne campylobacterosis in the USA was costing between $(US) 350-580. Hence, a reasonable conservative estimate is that each Australian case in 2010 would result in a cost of around $500 (this includes hospital, medication and lost productivity costs). Hence, this single food-borne agent could be costing Australian society around $115 million annually. In the light of these types of estimated costs for just one food-borne pathogen, it is easy to understand the importance that all food animal industries place on food safety.