Evaluation of ELISA coupled with Western blot as a surveillance tool for Trichinella infection in wild boar (Sus scrofa)

Trichinella surveillance in wildlife relies on muscle digestion of large samples which are logistically difficult to store and transport in remote and tropical regions as well as labour-intensive to process. Serological methods such as enzyme-linked immunosorbent assays (ELISAs) offer rapid, cost-effective alternatives for surveillance but should be paired with additional tests because of the high false-positive rates encountered in wildlife. We investigated the utility of ELISAs coupled with Western blot (WB) in providing evidence of Trichinella exposure or infection in wild boar. Serum samples were collected from 673 wild boar from a high- and low-risk region for Trichinella introduction within mainland Australia, which is considered Trichinella-free. Sera were examined using both an 'in-house' and a commercially available indirect-ELISA that used excretory secretory (E/S) antigens. Cut-off values for positive results were determined using sera from the low-risk population. All wild boar from the high-risk region (352) and 139/321 (43.3%) of the wild boar from the low-risk region were tested by artificial digestion. Testing by Western blot using E/S antigens, and a Trichinella-specific real-time PCR was also carried out on all ELISA-positive samples. The two ELISAs correctly classified all positive controls as well as one naturally infected wild boar from Gabba Island in the Torres Strait. In both the high- and low-risk populations, the ELISA results showed substantial agreement (k-value = 0.66) that increased to very good (k-value = 0.82) when WB-positive only samples were compared. The results of testing sera collected from the Australian mainland showed the Trichinella seroprevalence was 3.5% (95% C.I. 0.0-8.0) and 2.3% (95% C.I. 0.0-5.6) using the in-house and commercial ELISA coupled with WB respectively. These estimates were significantly higher (P < 0.05) than the artificial digestion estimate of 0.0% (95% C.I. 0.0-1.1). Real-time PCR testing of muscle from seropositive animals did not detect Trichinella DNA in any mainland animals, but did reveal the presence of a second larvae-positive wild boar on Gabba Island, supporting its utility as an alternative, highly sensitive method in muscle examination. The serology results suggest Australian wildlife may have been exposed to Trichinella parasites. However, because of the possibility of non-specific reactions with other parasitic infections, more work using well-defined cohorts of positive and negative samples is required. Even if the specificity of the ELISAs is proven to be low, their ability to correctly classify the small number of true positive sera in this study indicates utility in screening wild boar populations for reactive sera which can be followed up with additional testing. (C) 2013 Elsevier B.V. All rights reserved.

The effects of high stocking rates on milk production from dryland and irrigated Mediterranean pastures

An experiment using herds of similar to 20 cows (farmlets) assessed the effects of high stocking rates on production and profitability of feeding systems based on dryland and irrigated perennial ryegrass-based pastures in a Mediterranean environment in South Australia over 4 years. A target level of milk production of 7000 L/cow.year was set, based on predicted intakes of 2.7 t DM/cow.year as concentrates, pasture intakes from 1.5 to 2.7 t/cow.year and purchased fodder. In years 1 and 2, up to 1.5 t DM/cow.year of purchased fodder was used and in years 3 and 4 the amounts were increased if necessary to enable levels of milk production per cow to be maintained at target levels. Cows in dryland farmlets calved in March to May inclusive and were stocked at 2.5, 2.9, 3.3, 3.6 and 4.1 cows/ha, while those in irrigated farmlets calved in August to October inclusive and were stocked at 4.1, 5.2, 6.3 and 7.4 cows/ha. In the first 2 years, when inputs of purchased fodder were limited, milk production per cow was reduced with higher stocking rates (P < 0.01), but in years 3 and 4 there were no differences. Mean production was 7149 kg/cow.year in years 1 and 2, and 8162 kg/cow.year in years 3 and 4. Production per hectare was very closely related to stocking rate in all years (P < 0.01), increasing from 18 to 34 t milk/ha.year for dryland farmlets (1300 to 2200 kg milk solids/ha) and from 30 to 60 t milk/ha.year for irrigated farmlets (2200 to 4100 kg milk solids/ha). Almost all of these increases were attributed to the increases in grain and purchased fodder inputs associated with the increases in stocking rate. Net pasture accumulation rates and pasture harvest were generally not altered with stocking rate, though as stocking rate increased there was a change to more of the pasture being grazed and less conserved in both dryland and irrigated farmlets. Total pasture harvest averaged similar to 8 and 14 t DM/ha.year for dryland and irrigated pastures, respectively. An exception was at the highest stocking rate under irrigation, where pugging during winter was associated with a 14% reduction in annual pasture growth. There were several indications that these high stocking rates may not be sustainable without substantial changes in management practice. There were large and positive nutrient balances and associated increases in soil mineral content (P < 0.01), especially for phosphorus and nitrate nitrogen, with both stocking rate and succeeding years. Levels under irrigation were considerably higher (up to 90 and 240 mg/kg of soil for nitrate nitrogen and phosphorus, respectively) than under dryland pastures (60 and 140 mg/kg, respectively). Soil organic carbon levels did not change with stocking rate, indicating a high level of utilisation of forage grown. Weed ingress was also high (to 22% DM) in all treatments and especially in heavily stocked irrigated pastures during winter. It was concluded the higher stocking rates used exceeded those that are feasible for Mediterranean pastures in this environment and upper levels of stocking are suggested to be 2.5 cows/ha for dryland pastures and 5.2 cows/ha for irrigated pastures. To sustain these suggested stocking rates will require further development of management practices to avoid large increases in soil minerals and weed invasion of pastures.

A positive locality record or Lepidiota Contigua Britton (Coleoptera: Scarabaeidae: Melolonthinae)

Lepidiota contigua Britton is recorded from Windsor Tableland, northern Queensland.

Modelling Mortality and Reproduction Rates for Management of Sheep Flocks in Northern Australia

By quantifying the effects of climatic variability in the sheep grazing lands of north western and western Queensland, the key biological rates of mortality and reproduction can be predicted for sheep. These rates are essential components of a decision support package which can prove a useful management tool for producers, especially if they can easily obtain the necessary predictors. When the sub-models of the GRAZPLAN ruminant biology process model were re-parameterised from Queensland data along with an empirical equation predicting the probability of ewes mating added, the process model predicted the probability of pregnancy well (86% variation explained). Predicting mortality from GRAZPLAN was less successful but an empirical equation based on relative condition of the animal (a measure based on liveweight), pregnancy status and age explained 78% of the variation in mortalities. A crucial predictor in these models was liveweight which is not often recorded on producer properties. Empirical models based on climatic and pasture conditions estimated from the pasture production model GRASP, predicted marking and mortality rates for Mitchell grass (Astrebla sp.) pastures (81% and 63% of the variation explained). These prediction equations were tested against independent data from producer properties and the model successfully validated for Mitchell grass communities.

The effect of the cattle tick on growth rates and reproductive rates of Bos indicus cross heifers in north Queensland

Tick infestation occurs over 1.3 x 106 km2 in northern Australia. It has been difficult to estimate the economic effects of ticks due to a lack of information on their effects on growth and reproduction (Anon 1975). 12th Biennial Conference. February 1978. Melbourne, Victoria

Sperm production rates in Bos indicus strain bulls

Materials and Methods. Testes were collected a t castration or a t slaughter from purebred Brahman (B); Brahman cross (BX - half and three quarter); Sahiwal cross (SX – three quarter and seven eighths); and purebred and three quarter Santa Gertrudis (SG) bulls of known ages between 19 and 27 months and drawn from herds in northern coastal Queensland. 13th Biennial Conference. August 1980, Perth Western Australia.

Comparison of growth rates of relocated and local steers

Beef producers have expressed concern that cattle moved from one location to another do not always perform as well as comparable local cattle. Research station records and field trial data were examined to determine the effect of relocation on growth rate using data sets for animals of different age and liveweight at relocation and of different genotypes. 21st Biennial Conference. 8-12 July University of Queensland, Brisbane.

Phenotypic and genotypic characterisation of Pasteurella multocida isolated from pigs at slaughter in New Zealand

AIMS: To examine pigs at slaughter in New Zealand for the presence of Pasteurella multocida, and to determine for isolates, their biochemical profi les, somatic and capsular types, and the presence or absence of the HSB and toxA genes, associated with haemorrhagic septicaemia (HS) and progressive atrophic rhinitis (PAR), respectively. METHODS: Swabs from 173 lungs, 158 palatine tonsils and 82 nasal passages of pigs at two abattoirs in New Zealand were cultured for P. multocida using conventional techniques, and isolated colonies were subjected to biochemical tests for identi- fi cation of biovars. Somatic serotyping was conducted using an agar gel immunodiffusion (AGID) test. Polymerase chain reaction (PCR) assays were used to confi rm phenotypic identifi cation of colonies using species-specifi c primers, capsule type using serogroup-specifi c primers and multiplex PCR, and to test for the presence of HSB and toxA genes. RESULTS: Pasteurella multocida was isolated from 11/173 (6.4%) lung, 32/158 (20.2%) palatine tonsil and 5/82 (6.1 %) nasal swab samples, a total of 48 isolates from 413 samples (11.6%). Isolation rates per farm ranged from 1–53% of tissue samples collected from pigs 5–6 months of age. On phenotypic characterisation, isolates were allocated to seven main biovars, viz 1, 2, 3, 5, 9, 12, and a dulcitol-negative variant of Biovar 8, the majority (30/48) being Biovar 3. Of the 42 isolates for which somatic serotyping was conducted, 10% were Serovar 1, 79% were Serovar 3, 2% were Serovar 6,1, 2% were Serovar 12, and 7% could not be typed. All 48 isolates were confi rmed as P. multocida using a species-specifi c PCR. In the capsular multiplex PCR, 92% of isolates were Capsular (Cap) type A, 2% were Cap D, and 6% could not be typed. None of the samples were positive for the HSB or toxA genes. CONCLUSION: Serovars or capsular types of P. multocida associated with HS or PAR in pigs were not detected. Establishment of species-specifi c, capsular and toxin PCR assays allowed the rapid screening of isolates of P. multocida, while serotyping provided an additional tool for epidemiological and tracing purposes.

Quantitative assessment of total and Gram-positive aerobic bacteria in fresh and ambient-temperature-stored sub-tropical marine fish

The current study was undertaken to enumerate Gram-positive bacteria in fresh sub-tropical marine fish and determine the effect of ambient storage (25°C) on the Gram-positive bacterial count. Total and Gram-positive bacteria were enumerated in the muscles, gills and gut of fresh and stored Pseudocaranx dentex, Pagrus auratus and Mugil cephalus on tryptone soya agar (TSA) and TSA with 0.25% phenylethyl alcohol (PEA), respectively. Initial studies indicated that PEA significantly reduced total aerobic bacterial count (TABC) whereas control Gram-positive bacteria were not affected by 0.25% PEA. TABC significantly increased in all fish body parts, whereas Gram-positive aerobic bacterial count (GABC) significantly increased only in the muscles and gills during ambient storage for 15 h. The TABC of the fish species increased from 4.00, 6.13 and 4.58 log cfu g-1, respectively in the muscles, gills, and gut to 6.31, 7.31 and 7.23 log cfu g-1 by the end of storage. GABC increased from 2.00, 3.52 and 2.20 log cfu g-1 to 4.70, 5.85 and 3.36 log cfu g-1. Within each species, TABC were significantly higher in the gills compared to that of muscles and gut; however, no significant differences were found in GABC between muscles and gills. This study demonstrated the potential importance of Gram-positive bacteria in sub-tropical marine fish and their spoilage.

Growth and species interactions of Eucalyptus pellita in a mixed and monoculture plantation in the humid tropics of north Queensland

This study investigated whether mixed-species designs can increase the growth of a tropical eucalypt when compared to monocultures. Monocultures of Eucalyptus pellita (E) and Acacia peregrina (A) and mixtures in various proportions (75E:25A, 50E:50A, 25E:75A) were planted in a replacement series design on the Atherton Tablelands of north Queensland, Australia. High mortality in the establishment phase due to repeated damage by tropical cyclones altered the trial design. Effects of experimental designs on tree growth were estimated using a linear mixed-effects model with restricted maximum likelihood analysis (REML). Volume growth of individual eucalypt trees were positively affected by the presence of acacia trees at age 5 years and this effect generally increased with time up to age 10 years. However, the stand volume and basal area increased with increasing proportions of E. pellita, due to its larger individual tree size. Conventional analysis did not offer convincing support for mixed-species designs. Preliminary individual-based modelling using a modified Hegyi competition index offered a solution and an equation that indicates acacias have positive ecological interactions (facilitation or competitive reduction) and definitely do not cause competition like a eucalypt. These results suggest that significantly increased in growth rates could be achieved with mixed-species designs. This statistical methodology could enable a better understanding of species interactions in similarly altered experiments, or undesigned mixed-species plantations.

A universal test to determine the integrity of RNA, and its suitability for reverse transcription, in animal tissue laboratory specimens

Degradation of RNA in diagnostic specimens can cause false-negative test results and potential misdiagnosis when tests rely on the detection of specific RNA sequence. Current molecular methods of checking RNA integrity tend to be host species or group specific, necessitating libraries of primers and reaction conditions. The objective here was to develop a universal (multi-species) quality assurance tool for determining the integrity of RNA in animal tissues submitted to a laboratory for analyses. Ribosomal RNA (16S rRNA) transcribed from the mitochondrial 16S rDNA was used as template material for reverse transcription to cDNA and was amplified using polymerase chain reaction (PCR). As mitochondrial DNA has a high level of conservation, the primers used were shown to reverse transcribe and amplify RNA from every animal species tested. Deliberate degradation of rRNA template through temperature abuse of samples resulted in no reverse transcription and amplification. Samples spiked with viruses showed that single-stranded viral RNA and rRNA in the same sample degraded at similar rates, hence reverse transcription and PCR amplification of 16S rRNA could be used as a test of sample integrity and suitability for analysis that required the sample's RNA, including viral RNA. This test will be an invaluable quality assurance tool for determination of RNA integrity from tissue samples, thus avoiding erroneous test results that might occur if degraded target RNA is used unknowingly as template material for reverse transcription and subsequent PCR amplification.

Variable growth rates of the tropical estuarine fish barramundi Lates calcarifer (Bloch) under different freshwater flow conditions

Relationships between freshwater flows and growth rates of the opportunistic predatory finfish barramundi Lates calcarifer in a dry tropical estuary were examined using data from a long-term tag-recapture programme. Lagged effects were not investigated. After accounting for length at release, time at liberty and seasonal variation (e.g. winter, spring, summer and autumn), growth rates were significantly and positively related to fresh water flowing to the estuary. Effects were present at relatively low levels of freshwater flow (i.e. 2.15 m3 s-1, the 5th percentile of the mean flow rate experienced by fish in the study during time at liberty). The analysis, although correlative, provides quantitative evidence to support the hypothesis that freshwater flows are important in driving the productivity of estuaries and can improve growth of species high in the trophic chain.

Spatial and temporal variation and effects of changes in management in discard rates from the commercial reef line fishery of the Great Barrier Reef, Australia

Discarding in commercially exploited fisheries has received considerable attention in the last decade, though only more recently in Australia. The Reef Line fishery (RLF) of the Great Barrier Reef (GBR) in Australia is a large-scale multi-sector, multi-species, highly regulated hook and line fishery with the potential for high levels of discarding. We used a range of data sources to estimate discard rates and discard quantities for the two main target groups of the RLF, the coral trout, Plectropomus spp, and the red throat emperor, Lethrinus miniatus, and investigated possible effects on discarding of recent changes in management of the fishery. Fleet-wide estimates of total annual quantities discarded from 1989 to 2003 were 292-622 t and 33-95 t for coral trout and red throat emperor, respectively. Hypothetical scenarios of high-grading after the introduction of a total allowable commercial catch for coral trout resulted in increases in discard quantities up to 3895 t, while no high-grading still meant 421 t were discarded. Increasing the minimum size limit of red throat emperor from 35 to 38 cm also increased discards to an estimated 103 t. We provide spatially and temporally explicit estimates of discarding for the two most important species in the GBR RLF of Australia to demonstrate the importance of accounting for regional variation in quantification of discarding. Effects of management changes on discarding are also highlighted. This study provides a template for exploring discarding levels for other species in the RLF and elsewhere.

Simulating pasture growth rates in Australian and New Zealand grazing systems

DairyMod, EcoMod, and the SGS Pasture Model are mechanistic biophysical models developed to explore scenarios in grazing systems. The aim of this manuscript was to test the ability of the models to simulate net herbage accumulation rates of ryegrass-based pastures across a range of environments and pasture management systems in Australia and New Zealand. Measured monthly net herbage accumulation rate and accumulated yield data were collated from ten grazing system experiments at eight sites ranging from cool temperate to subtropical environments. The local climate, soil, pasture species, and management (N fertiliser, irrigation, and grazing or cutting pattern) were described in the model for each site, and net herbage accumulation rates modelled. The model adequately simulated the monthly net herbage accumulation rates across the range of environments, based on the summary statistics and observed patterns of seasonal growth, particularly when the variability in measured herbage accumulation rates was taken into account. Agreement between modelled and observed growth rates was more accurate and precise in temperate than in subtropical environments, and in winter and summer than in autumn and spring. Similarly, agreement between predicted and observed accumulated yields was more accurate than monthly net herbage accumulation. Different temperature parameters were used to describe the growth of perennial ryegrass cultivars and annual ryegrass; these differences were in line with observed growth patterns and breeding objectives. Results are discussed in the context of the difficulties in measuring pasture growth rates and model limitations.

Comparison of nitrogen fertilization methods and rates for subsurface drip irrigated corn in the semi-arid Great Plains

In semi-arid areas such as western Nebraska, interest in subsurface drip irrigation (SDI) for corn is increasing due to restricted irrigation allocations. However, crop response quantification to nitrogen (N) applications with SDI and the environmental benefits of multiple in-season (IS) SDI N applications instead of a single early-season (ES) surface application are lacking. The study was conducted in 2004, 2005, and 2006 at the University of Nebraska-Lincoln West Central Research and Extension Center in North Platte, Nebraska, comparing two N application methods (IS and ES) and three N rates (128, 186, and 278 kg N ha(-1)) using a randomized complete block design with four replications. No grain yield or biomass response was observed in 2004. In 2005 and 2006, corn grain yield and biomass production increased with increasing N rates, and the IS treatment increased grain yield, total N uptake, and gross return after N application costs (GRN) compared to the ES treatment. Chlorophyll meter readings taken at the R3 corn growth stage in 2006 showed that less N was supplied to the plant with ES compared to the IS treatment. At the end of the study, soil NO3-N masses in the 0.9 to 1.8 m depth were greater under the IS treatment compared to the ES treatment. Results suggested that greater losses of NO3-N below the root zone under the ES treatment may have had a negative effect on corn production. Under SDI systems, fertigating a recommended N rate at various corn growth stages can increase yields, GRN, and reduce NO3-N leaching in soils compared to concentrated early-season applications.