Gene expression profiling of cuticular proteins across the moult cycle of the crab Portunus pelagicus

Background: Crustaceans represent an attractive model to study biomineralization and cuticle matrix formation, as these events are precisely timed to occur at certain stages of the moult cycle. Moulting, the process by which crustaceans shed their exoskeleton, involves the partial breakdown of the old exoskeleton and the synthesis of a new cuticle. This cuticle is subdivided into layers, some of which become calcified while others remain uncalcified. The cuticle matrix consists of many different proteins that confer the physical properties, such as pliability, of the exoskeleton. Results: We have used a custom cDNA microarray chip, developed for the blue swimmer crab Portunus pelagicus, to generate expression profiles of genes involved in exoskeletal formation across the moult cycle. A total of 21 distinct moult-cycle related differentially expressed transcripts representing crustacean cuticular proteins were isolated. Of these, 13 contained copies of the cuticle_1 domain previously isolated from calcified regions of the crustacean exoskeleton, four transcripts contained a chitin_bind_4 domain (RR consensus sequence) associated with both the calcified and un-calcified cuticle of crustaceans, and four transcripts contained an unannotated domain (PfamB_109992) previously isolated from C. pagurus. Additionally, cryptocyanin, a hemolymph protein involved in cuticle synthesis and structural integrity, also displays differential expression related to the moult cycle. Moult stage-specific expression analysis of these transcripts revealed that differential gene expression occurs both among transcripts containing the same domain and among transcripts containing different domains. Conclusion: The large variety of genes associated with cuticle formation, and their differential expression across the crustacean moult cycle, point to the complexity of the processes associated with cuticle formation and hardening. This study provides a molecular entry path into the investigation of the gene networks associated with cuticle formation.

A multi-faceted approach for quantifying the estuarine-nearshore transition in the life cycle of the bull shark, Carcharhinus leucas.

Understanding the ontogenetic habitat linkages of sharks is important for conservation and managing human interactions. We used acoustic telemetry, catch data, elemental and stable isotope signatures and dietary analyses to investigate ontogenetic habitat use in south-east Queensland, Australia, by the bull shark Carcharhinus leucas, a IUCN 'near-threatened' species that is implicated in many shark attacks on humans in urban estuaries. Sequential analyses for delta(15)N and delta(13)C of vertebrae from five adult C. leucas and laser ablation inductively coupled plasma mass spectrometry (LA-ICPMS) for elemental composition from 23 C. leucas, including a pregnant female, were also used to trace ontogenetic habitat dependence. Acoustic telemetry indicated large juvenile and subadult C. leucas remained in estuarine habitats. delta(15)N values across shark vertebrae showed an ontogenetic shift in diet with total length (TL), confirmed by stomach contents. LA-ICPMS data reflected the ontogenetic movements of C. leucas from natal habitats. Differences among adults were gender related. Shifts in habitat use by subadults were correlated with a sigmoidal delta(13)C relationship with TL. C. leucas have a multipartite, stage-specific dependency in their transition between habitats along the freshwater-estuarine-marine continuum, making them particularly susceptible to the habitat alteration that is occurring globally.

Reproductive Biology and Spawning Periodicity of Endeavour Shrimps Metapenaeus endeavouri (Schmitt, 1926) and Metapenaeus ensis (de Haan, 1850) from a Central Queensland (Australia) Fishery

Metapenaeus endeavouri and M. ensis from coastal trawl fishing grounds off central Queensland, Australia, have marked seasonal reproductive cycles. Female M. endeavouri grew to a larger size than female M. ensis and occurred over a wider range of sites and depths. Although M. ensis was geographically restricted in distribution to only the shallowest sites it was highly abundant. Mating activity in these open thelycum species, indicated by the presence or absence of a spermatophore, was relatively low and highly seasonal compared with closed thelyeum shrimps. Seasonal variation in spermatophore insemination can be used as an independent technique to study spawning periodicity in open thelycum shrimps. Data strongly suggest an inshore movement of M. endeavouri to mature and spawn. This differs from most concepts of Penaeus species life cycles, but is consistent with the estuarine significance in the life cycle of Metapenaeus species. Monthly population fecundity indices suggest summer spawning for both species, which contrasts with the winter spawning of other shrimps from the same multispecies fishery.

Survival of Campylobacter spp. in darkling beetles (Alphitobius diaperinus) and their larvae in Australia

Campylobacter infection is the most frequently reported notifiable food-borne disease in humans in Australia. Our studies investigated the persistence of Campylobacter spp. in or on darkling beetles (Alphitobius diaperinus) and their larvae. Our results in analyses with chickens confirm that, unless very short turnaround times are used (<72 h), beetles colonized in one production cycle (i.e., one batch of chickens) are most unlikely to still be colonized during the next cycle of chickens.

Predicting risk and benefit a priori in biological control of invasive plant species: a systems modelling approach

While the method using specialist herbivores in managing invasive plants (classical biological control) is regarded as relatively safe and cost-effective in comparison to other methods of management, the rarity of strict monophagy among insect herbivores illustrates that, like any management option, biological control is not risk-free. The challenge for classical biological control is therefore to predict risks and benefits a priori. In this study we develop a simulation model that may aid in this process. We use this model to predict the risks and benefits of introducing the chrysomelid beetle Charidotis auroguttata to manage the invasive liana Macfadyena unguis-cati in Australia. Preliminary host-specificity testing of this herbivore indicated that there was limited feeding on a non-target plant, although the non-target was only able to sustain some transitions of the life cycle of the herbivore. The model includes herbivore, target and non-target life history and incorporates spillover dynamics of populations of this herbivore from the target to the non-target under a variety of scenarios. Data from studies of this herbivore in the native range and under quarantine were used to parameterize the model and predict the relative risks and benefits of this herbivore when the target and non-target plants co-occur. Key model outputs include population dynamics on target (apparent benefit) and non-target (apparent risk) and fitness consequences to the target (actual benefit) and non-target plant (actual risk) of herbivore damage. The model predicted that risk to the non-target became unacceptable (i.e. significant negative effects on fitness) when the ratio of target to non-target in a given patch ranged from 1:1 to 3:2. By comparing the current known distribution of the non-target and the predicted distribution of the target we were able to identify regions in Australia where the agent may be pose an unacceptable risk. By considering risk and benefit simultaneously, we highlight how such a simulation modelling approach can assist scientists and regulators in making more objective decisions a priori, on the value of releasing specialist herbivores as biological control agents.

The host range and biology of Cometaster pyrula; a biocontrol agent for Acacia nilotica subsp. indica in Australia

Prickly acacia, Acacia nilotica subsp. indica (Benth.) Brenan, a major weed of the Mitchell Grass Downs of northern Queensland, Australia, has been the target of biological control projects since the 1980s. The leaf-feeding caterpillar Cometaster pyrula (Hopffer) was collected from Acacia nilotica subsp. kraussiana (Benth.) Brenan during surveys in South Africa to find suitable biological control agents, recognised as a potential agent, and shipped into a quarantine facility in Australia. Cometaster pyrula has a life cycle of approximately 2 months during which time the larvae feed voraciously and reach 6 cm in length. Female moths oviposit a mean of 339 eggs. When presented with cut foliage of 77 plant species, unfed neonates survived for 7 days on only Acacia nilotica subsp. indica and Acacia nilotica subsp. kraussiana. When unfed neonates were placed on potted plants of 14 plant species, all larvae except those on Acacia nilotica subsp. indica and Acacia nilotica subsp. kraussiana died within 10 days of placement. Cometaster pyrula was considered to be highly host specific and safe to release in Australia. Permission to release C. pyrula in Australia was obtained and the insect was first released in north Queensland in October 2004. The ecoclimatic model CLIMEX indicated that coastal Queensland was climatically suitable for this insect but that inland areas were only marginally suitable.

Mechanically Ventilated Broiler Sheds: a Possible Source of Aerosolized Salmonella, Campylobacter, and Escherichia coli.

This study assessed the levels of two key pathogens, Salmonella and Campylobacter, along with the indicator organism Escherichia coli in aerosols within and outside poultry sheds. The study ranged over a 3-year period on four poultry farms and consisted of six trials across the boiler production cycle of around 55 days. Weekly testing of litter and aerosols was carried out through the cycle. A key point that emerged is that the levels of airborne bacteria are linked to the levels of these bacteria in litter. This hypothesis was demonstrated by E. coli. The typical levels of E. coli in litter were similar to 10(8) CFU g(-1) and, as a consequence, were in the range of 10(2) to 10(4) CFU m(-3) in aerosols, both inside and outside the shed. The external levels were always lower than the internal levels. Salmonella was only present intermittently in litter and at lower levels (10(3) to 10(5) most probable number [MPN] g(-1)) and consequently present only intermittently and at low levels in air inside (range of 0.65 to 4.4 MPN m(-3)) and once outside (2.3 MPN m(-3)). The Salmonella serovars isolated in litter were generally also isolated from aerosols and dust, with the Salmonella serovars Chester and Sofia being the dominant serovars across these interfaces. Campylobacter was detected late in the production cycle, in litter at levels of around 107 MPN g(-1). Campylobacter was detected only once inside the shed and then at low levels of 2.2 MPN m(-3). Thus, the public health risk from these organisms in poultry environments via the aerosol pathway is minimal.

Modeling population growth and site specific control of the invasive Lantana camara L. (Verbenaceae) under differing fire regimes

It is at the population level that an invasion either fails or succeeds. Lantana camara L. (Verbenaceae) is a weed of great significance in Queensland Australia and globally but its whole life-history ecology is poorly known. Here we used 3 years of field data across four land use types (farm, hoop pine plantation and two open eucalyptus forests, including one with a triennial fire regime) to parameterise the weed’s vital rates and develop size-structured matrix models. Lantana camara in its re-colonization phase, as observed in the recently cleared hoop pine plantation, was projected to increase more rapidly (annual growth rate, λ = 3.80) than at the other three sites (λ 1.88–2.71). Elasticity analyses indicated that growth contributed more (64.6 %) to λ than fecundity (18.5 %) or survival (15.5 %), while across size groups, the contribution was of the order: juvenile (19–27 %) ≥ seed (17–28 %) ≥ seedling (16–25 %) > small adult (4–26 %) ≥ medium adult (7–20 %) > large adult (0–20 %). From a control perspective it is difficult to determine a single weak point in the life cycle of lantana that might be exploited to reduce growth below a sustaining rate. The triennial fire regime applied did not alter the population elasticity structure nor resulted in local control of the weed. However, simulations showed that, except for the farm population, periodic burning could work within 4–10 years for control of the weed, but fire frequency should increase to at least once every 2 years. For the farm, site-specific control may be achieved by 15 years if the biennial fire frequency is tempered with increased burning intensity.

Biological control of Mikania micrantha in Papua New Guinea and Fiji using the rust fungus Puccinia spegazzinii

Mikania micrantha Kunth (Asteraceae), commonly known as ‘mile-a-minute’, is a neotropical plant species now found in 17 Pacific island countries and territories, invading small cropping areas and plantations, thereby reducing productivity and food security. In 2006, a biocontrol project on M. micrantha commenced in Fiji and Papua New Guinea (PNG). The distribution of M. micrantha as well as baseline data such as plant growth rates and socio-economic impacts were determined before the importation of any biocontrol agents. Mikania micrantha was recorded in all 15 lowland provinces in PNG and on all major islands in Fiji. Plants grow about 3.2cm/day in PNG and about 1.9cm/day in Fiji. A socio-economic survey, involving over 370 respondents in over 220 villages from 15 provinces in PNG, found that 78% of respondents considered M. micrantha a serious weed and about 44% had M. micrantha, which they needed to weed at least fortnightly, in over a third of their land. Over 80% of respondents used slashing and/or handpulling as the preferred method of weed control. About 40% of respondents considered that M. micrantha reduced crop yield by more than 30%. In Fiji, 52 respondents from four islands participated in the survey. Over 60% of respondents in Fiji considered M. micrantha a serious weed and 23% had about 30% of their farm lands infested with the weed. Only 15% of respondents needed to weed at least fortnightly, with 56% using slashing and/or hand-pulling as the preferred means of control. Over 65% of respondents estimated that they lost at least 30% of potential crop yield to M. micrantha. Nearly 90% of respondents used M. micrantha as a medicinal plant to treat cuts and wounds. The life history of the rust Puccinia spegazzinii de Toni (Pucciniales: Pucciniaceae), originating from Ecuador, and imported into PNG and Fiji in 2008, was studied. P. spegazzinii is a microcyclic and autoecious rust and has a life cycle of 18-22 days. An efficient culturing and field release method was developed. Since 2008, the rust has been released at over 450 sites in 15 provinces in PNG, establishing at nearly 70 sites in four provinces. From some sites, the rust has spread over 7 km in 12 months. In Fiji, the rust has been released at over 80 sites, on four of the main islands, namely Viti Levu, Vanua Levu, Taveuni and Ovalau, and has established at 20 sites on Viti Levu and Vanua Levu. Plant growth studies and field monitoring in PNG showed that P. spegazzinii can significantly reduce the growth and density of M. micrantha and offers great potential for the control of this weed.

Novel vaccine acceptance in emerging infectious diseases: a case study of Hendra virus vaccine uptake in Australia

Background: The development of a horse vaccine against Hendra virus has been hailed as a good example of a One Health approach to the control of human disease. Although there is little doubt that this is true, it is clear from the underwhelming uptake of the vaccine by horse owners to date (approximately 10%) that realisation of a One Health approach requires more than just a scientific solution. As emerging infectious diseases may often be linked to the development and implementation of novel vaccines this presentation will discuss factors influencing their uptake; using Hendra virus in Australia as a case study. Methods: This presentation will draw on data collected from the Horse owners and Hendra virus: A Longitudinal cohort study To Evaluate Risk (HHALTER) study. The HHALTER study is a mixed methods research study comprising a two-year survey-based longitudinal cohort study and qualitative interview study with horse owners in Australia. The HHALTER study has investigated and tracked changes in a broad range of issues around early uptake of vaccination, horse owner uptake of other recommended disease risk mitigation strategies, and attitudes to government policy and disease response. Interviews provide further insights into attitudes towards risk and decision-making in relation to vaccine uptake. A combination of quantitative and qualitative data analysis will be reported. Results: Data collected from more than 1100 horse owners shortly after vaccine introduction indicated that vaccine uptake and intention to vaccinate was associated with a number of risk perception factors and financial cost factors. In addition, concerns about side effects and veterinarians refusing to treat unvaccinated horses were linked to uptake. Across the study period vaccine uptake in the study cohort increased to more than 50%, however, concerns around side effects, equine performance and breeding impacts, delays to full vaccine approvals, and attempts to mandate vaccination by horse associations and event organisers have all impacted acceptance. Conclusion: Despite being provided with a safe and effective vaccine for Hendra virus that can protect horses and break the transmission cycle of the virus to humans, Australian horse owners have been reluctant to commit to it. General issues pertinent to novel vaccines, combined with challenges in the implementation of the vaccine have led to issues of mistrust and misconception with some horse owners. Moreover, factors such as cost, booster dose schedules, complexities around perceived risk, and ulterior motives attributed to veterinarians have only served to polarise attitudes to vaccine acceptance.

Developmental biology and prey preference of Diomus notescens Blackburn (Coleoptera: Coccinellidae): A predator of Aphis gossypii Glover (Hemiptera: Aphididae)

The minute two-spotted ladybeetle, Diomus notescens Blackburn is a common predator of aphids and other pests in Australian agricultural crops, however little is known about the biology of D. notescens. The aim of this study was to provide information on the life cycle of this predator and improve our understanding of its biological control potential, particularly against one of the major pests of cotton, Aphis gossypii Glover. In laboratory experiments, juvenile development, prey consumption, as well as adult lifespan and fecundity were studied. Results from this study revealed that D. notescens could successfully complete development on A. gossypii, which at 25 °C required 21 days and during this period they each consume 129 ± 5.2 aphids. At 25 °C adult lifespan was 77 ± 9.6 days, with a mean daily prey consumption of 28 ± 1.8 aphids and a mean daily fecundity of 8 ± 0.5 eggs. Net reproductive rate was estimated as 187 ± 25.1 females and the intrinsic rate of increase was estimated as 0.14. Juvenile development was recorded at four constant temperatures (15, 21, 26 and 27 °C) and using a linear model, the lower threshold for D. notescens development was estimated to be 10 ± 0.6 °C with 285 ± 4.7 degree days required to complete development. A prey choice experiment studying predation rates revealed a strong preference for A. gossypii nymphs compared to Bemisia tabaci Gennadius eggs.