A Genetic Analysis of Lysosomal Enzyme Activities in Brahman Cattle

Analyses of variance and co variance were carried out on the activities of three lysosomal enzymes in mononuclear blood cells from Brahman cattle. These were hexosaminidase (HEX), beta-D-galacto-sidase (GAL) and acid alpha-glucosidase (GLU) which had been measured in blood mononuclear cells from 1752 cattle from 6 herds in a Pompe's disease control programme. Herd of origin and date of bleeding significantly affected the level of activity of all enzymes. In addition, HEX and GAL were affected by age and HEX by the sex of the animal bled. Estimates of heritability from sire variances were 0.29:t 0.09 for HEX, 0.31 :t 0.09 for GAL and 0.44:t 0.09 for GLU. Genetic correlations between all enzymes were positive. The data indicate the existence of a major gene causing Pompe's disease and responsible for 16% of the genetic variation in GLU. One standard deviation of selection differential for high GLU should almost eliminate Pompe's disease from the population. The effi-ciency of selection would be aided by estimating the breeding value for GLU using measurements of HEX and GLU and taking account of an animal's sex, age, date of bleeding and herd of origin.

Validation of a Competitive Enzyme-Linked Immunosorbent Assay for Detection of Babesia bigemina Antibodies in Cattle

A competitive enzyme-linked immunosorbent assay (cELISA) based on a broadly conserved, species-specific, B-cell epitope within the C terminus of Babesia bigemina rhoptry-associated protein 1a was validated for international use. Receiver operating characteristic analysis revealed 16% inhibition as the threshold for a negative result, with an associated specificity of 98.3% and sensitivity of 94.7%. Increasing the threshold to 21% increased the specificity to 100% but modestly decreased the sensitivity to 87.2%. By using 21% inhibition, the positive predictive values ranged from 90.7% (10% prevalence) to 100% (95% prevalence) and the negative predictive values ranged from 97.0% (10% prevalence) to 48.2% (95% prevalence). The assay was able to detect serum antibody as early as 7 days after intravenous inoculation. The cELISA was distributed to five different laboratories along with a reference set of 100 defined bovine serum samples, including known positive, known negative, and field samples. The pairwise concordance among the five laboratories ranged from 100% to 97%, and all kappa values were above 0.8, indicating a high degree of reliability. Overall, the cELISA appears to have the attributes necessary for international application.

Evaluation of partial replacement of live and fresh feeds with a formulated diet and the influence of weaning Panulirus ornatus phyllosomata onto a formulated diet during early ontogeny

We have evaluated the potential of a formulated diet as a replacement for live and fresh feeds for 7-day post-hatch Panulirus ornatus phyllosomata and also investigated the effect of conditioning phyllosomata for 14-21 days on live feeds prior to weaning onto a 100% formulated diet. In the first trial, the highest survival (>55%) was consistently shown by phyllosomata fed a diet consisting of a 50% combination of Artemia nauplii and 50% Greenshell mussel, followed by phyllosomata fed 50% Artemia nauplii and 50% formulated diet and, thirdly, by those receiving 100% Artemia nauplii. The second trial assessed the replacement of on-grown Artemia with proportions of formulated diet and Greenshell mussel that differed from those used in trial 1. Phyllosomata fed a 75% combination of formulated diet and 25% on-grown Artemia and 50% on-grown Artemia and 50% Greenshell mussel consistently showed the highest survival (>75%). Combinations of Greenshell mussel and formulated diet resulted in significantly (P < 0.05) reduced survival. In trial 3, phyllosomata were conditioned for 14, 18 or 21 days on Artemia nauplii prior to weaning onto a 100% formulated diet, which resulted in survival rates that were negatively related to the duration of feeding Artemia nauplii. In the final trial, phyllosomata were conditioned for 14 days on live on-grown Artemia prior to weaning onto one of three formulated diets (one diet with 44% CP and two diets with 50%). Phyllosomata fed a 44% CP diet consistently showed the highest survival (>35%) among all treatments, while those fed a 50%-squid CP diet showed a significant (P < 0.05) increase in mortality at day 24. The results of these trials demonstrate that hatcheries can potentially replace 75% of live on-grown Artemia with a formulated diet 7 days after hatch. The poor performance associated with feeding combinations of Greenshell mussel and formulated diet, and 100% formulated diet as well as conditioning phyllosomata for 14-21 days on live feeds prior to weaning onto a formulated diet highlights the importance of providing Artemia to stimulate feeding.

A free-enzyme catalyst for the bioremediation of environmental atrazine contamination.

Herbicide contamination from agriculture is a major issue worldwide, and has been identified as a threat to freshwater and marine environments in the Great Barrier Reef World Heritage Area in Australia. The triazine herbicides are of particular concern because of potential adverse effects, both on photosynthetic organisms and upon vertebrate development. To date a number of bioremediation strategies have been proposed for triazine herbicides, but are unlikely to be implemented due to their reliance upon the release of genetically modified organisms. We propose an alternative strategy using a free-enzyme bioremediant, which is unconstrained by the issues surrounding the use of live organisms. Here we report an initial field trial with an enzyme-based product, demonstrating that the technology is technically capable of remediating water bodies contaminated with the most common triazine herbicide, atrazine.

Archaeaphage therapy to control rumen methanogens

Phage therapy is becoming increasingly important as a means of eradicating or controlling microbial populations and has been raised as a potential strategy to reduce methane emissions from ruminants. To date, very little is currently known about phages which may infect the methane-producing archaeal strains (methanogens) dominant within the rumen of Australian cattle, such as the Methanobrevibacter ruminantium. This project aimed to assemble a collection of phages to be employed in phage therapy. A range of animal-derived and environmental source samples were tested using culture-based methodology, however no lytic phages of methanogens were isolated. Given the dearth of knowledge regarding phages of rumen methanogens, this project established that these naturally-occurring phages may be present in very low concentrations within the rumen and this will need to be considered in future methanogen-phage isolation investigations. The project has begun the process of developing and adapting new methodologies for detecting and examining these phages

Early season replacement for Imperial Mandarin CT09014

Imperial mandarin is the dominant variety in the early season production time slot in Australia. It is well known to consumers, wholesalers and supermarkets, and growers continue to make new plantings of this variety. Despite this popularity, Imperial has major faults that cause problems for growers, consumers, and at other points along the supply chain. Fruit of the variety can be severely granulated, and this is the main quality problem cited by consumers. A second problem with Imperial is its unsuitability for export. Thirdly, the variety has low juice and sugar content, and develops poor colour under warm conditions. We propose a hybridisation project to specifically address these industry obstacles.

A core metabolic enzyme mediates resistance to phosphine gas

Phosphine is a small redox-active gas that is used to protect global grain reserves, which are threatened by the emergence of phosphine resistance in pest insects. We find that polymorphisms responsible for genetic resistance cluster around the redox-active catalytic disulfide or the dimerization interface of dihydrolipoamide dehydrogenase (DLD) in insects (Rhyzopertha dominica and Tribolium castaneum) and nematodes (Caenorhabditis elegans). DLD is a core metabolic enzyme representing a new class of resistance factor for a redox-active metabolic toxin. It participates in four key steps of core metabolism, and metabolite profiles indicate that phosphine exposure in mutant and wild-type animals affects these steps differently. Mutation of DLD in C. elegans increases arsenite sensitivity. This specific vulnerability may be exploited to control phosphine-resistant insects and safeguard food security.

Phage therapy in livestock methane amelioration

Viruses of prokaryotes (phages) are obligate microbial pathogens that can, in the lytic phase of development, infect and lyse their respective bacterial or archaeal hosts. As such, these viruses can reduce the population density of their hosts rapidly, and have been viewed as possible agents of biological control (phage therapy). Phage therapy is becoming increasingly important as a means of eradicating or controlling microbial populations as the use of antibiotics and chemical treatments becomes both less effective and less publicly acceptable. Phage therapy has therefore been raised as a potential strategy to reduce methane (CH 4) emissions from ruminants, providing an innovative biological approach, harnessing the potent, yet targeted, biocidal attributes of these naturally occurring microbial predators.

Phage therapy in livestock methane amelioration.

Viruses of prokaryotes (phages) are obligate microbial pathogens that can, in the lytic phase of development, infect and lyse their respective bacterial or archaeal hosts. As such, these viruses can reduce the population density of their hosts rapidly, and have been viewed as possible agents of biological control (phage therapy). Phage therapy is becoming increasingly important as a means of eradicating or controlling microbial populations as the use of antibiotics and chemical treatments becomes both less effective and less publicly acceptable. Phage therapy has therefore been raised as a potential strategy to reduce methane (CH4) emissions from ruminants, providing an innovative biological approach, harnessing the potent, yet targeted, biocidal attributes of these naturally occurring microbial predators.

Fish Silage as Replacement of Fishmeal in Red Tilapia Feeds

We estimated the effect on growth and nutrient efficiency of replacing fishmeal with silage incorporated with rice bran in diets for fingerling red tilapia (Oreochromis mossambicus × Oreochromis niloticus × Oreochromis aureus) over 12 weeks. Isonitrogenous (300 g kg−1 protein dry matter basis) and isoenergetic (4450 Kcal gross energy kg−1) feed formulations with increasing levels of tilapia silage as a replacement for fishmeal were prepared: Diet 1 with no silage (0 g Kg−1), Diet 2 (250 g Kg−1), Diet 3 (500 g Kg−1), and Diet 4 (750 g Kg−1). Feed intake was similar among Diets 1, 2, and 3, while Diet 4 had a significantly lower intake. There was no significant difference (P > 0.05) in weight gain or specific growth rate (SGR), feed conversion ratio (FCR), and protein efficiency ratio (PER), among fish fed Diets 1, 2, and 3. Fish fed with Diet 4 had significantly lower weight gain; SGR and PER and significantly higher FCR. Organoleptic properties of the fish were not affected by the diets. The results of this study indicate that less expensive dried fish silage with rice bran is an alternative protein source for tilapia feed up to 50% of fishmeal replacement.