Fine tuning for the tropics: application of eDNA technology for invasive fish detection in tropical freshwater ecosystems

Invasive species pose a major threat to aquatic ecosystems. Their impact can be particularly severe in tropical regions, like those in northern Australia, where >20 invasive fish species are recorded. In temperate regions, environmental DNA (eDNA) technology is gaining momentum as a tool to detect aquatic pests, but the technology's effectiveness has not been fully explored in tropical systems with their unique climatic challenges (i.e. high turbidity, temperatures and ultraviolet light). In this study, we modified conventional eDNA protocols for use in tropical environments using the invasive fish, Mozambique tilapia (Oreochromis mossambicus) as a detection model. We evaluated the effects of high water temperatures and fish density on the detection of tilapia eDNA, using filters with larger pores to facilitate filtration. Large-pore filters (20 μm) were effective in filtering turbid waters and retaining sufficient eDNA, whilst achieving filtration times of 2-3 min per 2-L sample. High water temperatures, often experienced in the tropics (23, 29, 35 °C), did not affect eDNA degradation rates, although high temperatures (35 °C) did significantly increase fish eDNA shedding rates. We established a minimum detection limit for tilapia (1 fish/0.4 megalitres/after 4 days) and found that low water flow (3.17 L/s) into ponds with high fish density (>16 fish/0.4 megalitres) did not affect eDNA detection. These results demonstrate that eDNA technology can be effectively used in tropical ecosystems to detect invasive fish species. © 2016 John Wiley & Sons Ltd.

Identification of genes involved with tick infestation in Bos taurus and Bos indicus

Tick resistant cattle could provide a potentially sustainable and environmentally sound method of controlling cattle ticks. Advances in genomics and the availability of the bovine genome sequence open up opportunities to identify useful and selectable genes controlling cattle tick resistance. Using quantitative real-time PCR and the Affymetrix bovine array platform, differences in gene expression of skin biopsies from tick resistant Bos indicus (Brahman) and tick susceptible Bos taurus (Holstein-Friesian) cattle following tick challenge were examined. We identified 138 significant differentially-expressed genes, including several immunological/host defence genes, extracellular matrix proteins, and transcription factors as well as genes involved in lipid metabolism. Three key pathways, represented by genes differentially expressed in resistant Brahmans, were identified; the development of the cell-mediated immune response, structural integrity of the dermis and intracellular Ca 2+ levels. Ca2+, which is implicated in host responses to microbial stimuli, may be required for the enhancement or fine-tuning of transcriptional activation of Ca2+- dependant host defence signalling pathways. Animal Genomics for Animal Health International Symposium, Paris, October 2007: (Proceedings)

Variability in odour reception in the peripheral sensory system of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

Mixtures of single odours were used to explore the receptor response profile across individual antennae of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae). Seven odours were tested including floral and green-leaf volatiles: phenyl acetaldehyde, benzaldehyde, b-caryophyllene, limonene, a-pinene, 1-hexanol, 3Z-hexenyl acetate. Electroantennograms of responses to paired mixtures of odours showed that there was considerable variation in receptor tuning across the receptor field between individuals. Data from some moth antennae showed no additivity, which indicated a restricted receptor profile. Results from other moth antennae to the same odour mixtures showed a range of partial additivity. This indicated that a wider array of receptor types was present in these moths, with a greater percentage of the receptors tuned exclusively to each odour. Peripheral receptor fields show variation in the spectrum of response within a population (of moths) when exposed to high doses of plant volatiles. This may be related to recorded variation in host choice within moth populations as reported by other authors.

Summative evaluation of climate application activities with pastoralists in western Queensland

Survey methods were engaged to measure the change in use and knowledge of climate information by pastoralists in western Queensland. The initial mail survey was undertaken in 2000-01 (n=43) and provided a useful benchmark of pastoralists climate knowledge. Two years of climate applications activities were completed and clients were re-surveyed in 2003 (n=49) to measure the change in knowledge and assess the effectiveness of the climate applications activities. Two methods were used to assess changes in client knowledge, viz., self-assessment and test questions. We found that the use of seasonal climate forecasts in decision making increased from 36% in 2001 (n=42) to 51% in 2003 (n=49) (P=0.07). The self-assessment technique was unsatisfactory as a measure of changing knowledge over short periods (1-3 years), but the test question technique was successful and indicated an improvement in climate knowledge among respondents. The increased levels of use of seasonal climate forecasts in management and improved knowledge was partly attributed to the climate applications activities of the project. Further, those who used seasonal forecasting (n=25) didn't understand key components of forecasts (e.g. probability, median) better than those who didn't use seasonal forecasts (n=24) (P>0.05). This identifies the potential for misunderstanding and misinterpretation of forecasts among users and highlights the need for providers of forecasts to understand the difficulties and prepare simply written descriptions of forecasts and disseminate these with the maps showing probabilities. The most preferred means of accessing climate information were internet, email, 'The Season Ahead' newsletter and newspaper. The least preferred were direct contact with extension officers and attending field days and group meetings. Eighty-six percent of respondents used the internet and 67% used ADSL broadband internet (April 2003). Despite these findings, extension officers play a key role in preparing and publishing the information on the web, in emails and newsletters. We also believe that direct contact with extension officers trained in climate applications is desirable in workshop-like events to improve knowledge of the difficult concepts underpinning climate forecasts, which may then stimulate further adoption.

Increased reproductive success in older female red kangaroos and the impact of harvesting

Wildlife harvesting has a long history in Australia, including obvious examples of overexploitation. Not surprisingly, there is scepticism that commercial harvesting can be undertaken sustainably. Kangaroo harvesting has been challenged regularly at Administrative Appeals Tribunals and elsewhere over the past three decades. Initially, the concern from conservation groups was sustainability of the harvest. This has been addressed through regular, direct monitoring that now spans > 30 years and a conservative harvest regime with a low risk of overharvest in the face of uncertainty. Opposition to the harvest now continues from animal rights groups whose concerns have shifted from overall harvest sustainability to side effects such as animal welfare, and changes to community structure, genetic composition and population age structure. Many of these concerns are speculative and difficult to address, requiring expensive data. One concern is that older females are the more successful breeders and teach their daughters optimal habitat and diet selection. The lack of older animals in a harvested population may reduce the fitness of the remaining individuals; implying population viability would also be compromised. This argument can be countered by the persistence of populations under harvesting without any obvious impairment to reproduction. Nevertheless, an interesting question is how age influences reproductive output. In this study, data collected from a number of red kangaroo populations across eastern Australia indicate that the breeding success of older females is up to 7-20% higher than that of younger females. This effect is smaller than that of body condition and the environment, which can increase breeding success by up to 30% and 60% respectively. Average age of mature females in a population may be reduced from 9 to 6 years old, resulting in a potential reduction in breeding success of 3-4%. This appears to be offset in harvested populations by improved condition of females from a reduction in kangaroo density. There is an important recommendation for management. The best insurance policy against overharvest and unwanted side effects is not research, which could be never-ending. Rather, it is a harvest strategy that includes safeguards against uncertainty such as harvest reserves, conservative quotas and regular monitoring. Research is still important in fine tuning that strategy and is most usefully incorporated as adaptive management where it can address the key questions on how populations respond to harvesting.

Organic diet development for black tiger prawns

The survival and growth of black tiger prawn (Penaeus monodon) juveniles (~3.3 g) were compared after feeding in tanks over one month with several prepared diets based on organically certified ingredients. The extrusion process in the manufacture of pelletised experimental diets was similar to processes used in commercial plants and was closely documented. The daily feeding rate (6% of starting mean body weight) was split equally into two feeds, one in the morning and one in the afternoon. All diets tested produced high survival (97-100%). A widely-used commercial Australian prawn feed was used as a control diet. It contained 41.2% protein with 29.5 g kg-1 lysine, and produced the highest (P<0.05) growth (117% weight gain). Three of the experimental organic diets tested (namely, 1. wheat + soy, 2. pig weaner diet + soy, and 3. pig weaner diet + dried fish waste) produced moderate growth (73–77% weight gain). These contained 33%, 36% or 31% protein, respectively, and produced better (P<0.05) growth than diets utilising a range of other prospective ingredients (eg: wheat + dried scallop gut, wheat + fish waste, wheat + chickpea, or wheat + macadamia meal, containing 23%, 25%, 29% or 24% protein, respectively). An unfed control-treatment produced the lowest (P<0.05) growth (4% weight gain). The water stability of the experimental diets that produced the best growth was poorer than the commercial diet, suggesting that improvements in this aspect of these organic feed’s manufacture could result in additional performance benefits and possibly reduced feed wastage. Analyses revealed a linear relationship between diet performance (in terms of weight gains) and the protein and lysine contents of diets. About 70% of diet performance was explained by these factors. The superior performance of the commercial diet could be attributed primarily to its formulation using mainly marine proteins, as well as a range of other unknown factors (commercial in confidence). These other factors range from use of feed attractants, better knowledge of ingredient nutrient availability, different extrusion conditions and the use of other unspecified micro-nutrients not present in the experimental diets. The organic diets studied still require a degree of fine-tuning before structured commercial uptake. This would sensibly include further detailed investigations of the composition and nutrient availabilities of these and other organic dietary ingredients, and refinement of the extrusion process for formulated diets.

Synthesis of Silica Vesicles with Controlled Entrance Size for High Loading, Sustained Release, and Cellular Delivery of Therapeutical Proteins

A rationally designed two-step synthesis of silica vesicles is developed with the formation of vesicular structure in the first step and fine control over the entrance size by tuning the temperature in the second step. The silica vesicles have a uniform size of ≈50 nm with excellent cellular uptake performance. When the entrance size is equal to the wall thickness, silica vesicles after hydrophobic modification show the highest loading amount (563 mg/g) towards Ribonuclease A with a sustained release behavior. Consequently, the silica vesicles are excellent nano-carriers for cellular delivery applications of therapeutical biomolecules.