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.

Multiplex real-time PCR for the detection and differentiation of equid herpesvirus 1 (EHV-1) and equid herpesvirus 4 (EHV-4).

A multiplex real-time PCR was designed to detect and differentiate equid herpesvirus 1 (EHV-1) and equid herpesvirus 4 (EHV-4). The PCR targets the glycoprotein B gene of EHV-1 and EHV-4. Primers and probes were specific to each equine herpesvirus type and can be used in monoplex or multiplex PCRs, allowing the differentiation of these two closely related members of the Alphaherpesvirinae. The two probes were minor-groove binding probes (MGB?) labelled with 6-carboxy-fluorescein (FAM?) and VIC® for detection of EHV-1 and EHV-4, respectively. Ten EHV-1 isolates, six EHV-1 positive clinical samples, one EHV-1 reference strain (EHV-1.438/77), three EHV-4 positive clinical samples, two EHV-4 isolates and one EHV-4 reference strain (EHV-4 405/76) were included in this study. EHV-1 isolates, clinical samples and the reference strain reacted in the EHV-1 real-time PCR but not in the EHV-4 real-time PCR and similarly EHV-4 clinical samples, isolates and the reference strain were positive in the EHV-4 real-time PCR but not in the EHV-1 real-time PCR. Other herpesviruses, such as EHV-2, EHV-3 and EHV-5 were all negative when tested using the multiplex real-time PCR. When bacterial pathogens and opportunistic pathogens were tested in the multiplex real-time PCR they did not react with either system. The multiplex PCR was shown to be sensitive and specific and is a useful tool for detection and differentiation of EHV-1 and EHV-4 in a single reaction. A comprehensive equine herpesvirus disease investigation procedure used in our laboratory is also outlined. This procedure describes the combination of alphaherpesvirus multiplex real-time PCR along with existing gel-based PCRs described by other authors.

The efficacy of a vitamin D3 metabolite for improving the myofibrillar tenderness of meat from Bos indicus cattle

The influence of a once only administration of a metabolite of vitamin D3 (HY [middle dot] D(R)-25-hydroxy vitamin D3) on myofibrillar meat tenderness in Australian Brahman cattle was studied. Ninety-six Brahman steers of three phenotypes (Indo-Brazil, US and US/European) and with two previous hormonal growth promotant (HGP) histories (implanted or not implanted with Compudose(R)) were fed a standard feedlot ration for 70 d. Treatment groups of 24 steers were offered daily 10 g/head HY [middle dot] D(R) (125 mg 25-hydroxyvitamin D3) for 6, 4, or 2 d before slaughter. One other group of 24 steers was given the basal diet without HY [middle dot] D(R). Feed lot performance, blood and muscle samples and carcass quality data were collected at slaughter. Calcium, magnesium, potassium, sodium, iron and Vitamin D3 metabolites were measured in plasma and longissimus dorsi muscle. Warner-Bratzler (WB) shear force (peak force, initial yield) and other objective meat quality measurements were made on the longissimus dorsi muscle of each steer after ageing for 1, 7 and 14 d post-mortem at 0-2 [deg]C.There were no significant effects of HY [middle dot] D(R) supplements on average daily gain (ADG, 1.28-1.45 kg/d) over the experimental period. HY [middle dot] D(R) supplements given 6 d prior to slaughter resulted in significantly higher (P (R)) by phenotype/HGP interaction for peak force (P = 0.028), in which Indo-Brazil steers without previous HGP treatment responded positively (increased tenderness) to HY [middle dot] D(R) supplements at 2 d when compared with Indo-Brazil steers previously given HGP. There were no significant effects of treatment on other phenotypes. HY [middle dot] D(R) supplements did not affect muscle or plasma concentrations of calcium, potassium or sodium, but did significantly decrease plasma magnesium and iron concentrations when given 2 d before slaughter. There were no detectable amounts of 25-hydroxyvitamin D3 in the blood or muscle of any cattle at slaughter.

Characterization of tree-to-tree variation in morphological, nutritional and medicinal properties of Canarium indicum nuts

As part of a feasibility study of the commercialization potential of C. indicum nuts as Agroforestry Tree Products in Papua New Guinea, preliminary characterization studies have examined the tree-to-tree variation in morphological traits (nut and kernel mass and kernel:nut ratio), as well as nutritional (carbohydrate, fat, protein, sodium, vitamin E) and medicinal traits (anti-oxidant activity, anti-inflammatory activity and phenolic content) of kernels from 18 to 72 trees in a small number of different villages of Papua New Guinea (East New Britain Province). There was continuous variation in these traits indicating opportunities for multiple trait cultivar development targeted at food and pharmaceutical markets. Certain traits, for example anti-inflammatory activity, in which tree-to-tree variation was highly significant, present greater opportunities than others, such as saturated:unsaturated fatty acid ratio. This intraspecific variation was greater within populations than between populations. The data presented has allowed the development of a strategy to domesticate C. indicum for cultivation in homegardens and cocoa-coconut agroforests, using a participatory approach aimed at the production of agroforestry tree products (AFTPs) to empower small-holders and enhance their livelihoods and income.

Rainfall and streamflow response to El Niño Southern Oscillation: a case study in a semiarid catchment, Australia.

This paper aims to compare the shift in frequency distribution and skill of seasonal climate forecasting of both streamflow and rainfall in eastern Australia based on the Southern Oscillation Index (SOI) Phase system. Recent advances in seasonal forecasting of climate variables have highlighted opportunities for improving decision making in natural resources management. Forecasting of rainfall probabilities for different regions in Australia is available, but the use of similar forecasts for water resource supply has not been developed. The use of streamflow forecasts may provide better information for decision-making in irrigation supply and flow management for improved ecological outcomes. To examine the relative efficacy of seasonal forecasting of streamflow and rainfall, the shift in probability distributions and the forecast skill were evaluated using the Wilcoxon rank-sum test and the linear error in probability space (LEPS) skill score, respectively, at three river gauging stations in the Border Rivers Catchment of the Murray-Darling Basin in eastern Australia. A comparison of rainfall and streamflow distributions confirms higher statistical significance in the shift of streamflow distribution than that in rainfall distribution. Moreover, streamflow distribution showed greater skill of forecasting with 0-3 month lead time, compared to rainfall distribution.

A blowfly strike vaccine requires an understanding of host-pathogen interactions

The phase-out of Mulesing by 2010 means the Australian wool industry requires immediate and viable alternatives for the control and prevention of blowfly strike, an economically important parasitic disease of sheep. In this review we have analysed previous research aimed toward the development of a vaccine against blowfly strike and the reasons why the approaches taken were unsuccessful at the time. Close scrutiny has provided new insight into this host-parasite interaction and identified new opportunities for the development of a vaccine. Here we propose that addressing immunosuppression together with the induction of cellular immunity is likely to result in an anti-blowfly strike vaccine, as opposed to the use of "standard" approaches aimed at inducing humoral immunity.

Carotenoid and vitamin content of Karat and other Micronesian banana cultivars.

We previously found high carotenoid levels in Karat and other Micronesian bananas, indicating potential importance for alleviating vitamin A deficiency and other nutritionally related health problems in the Federated States of Micronesia. Past work focused on carotenoid and mineral analyses, whereas here we investigated 16 cultivars (most not previously analysed) for a broader micronutrient profile, including seven vitamins. Karat carotenoid levels were higher than in previous analyses, confirming Karat as exceptionally carotenoid-rich. We identified an additional 10 carotenoid-rich cultivars, expanding the range having potential for alleviating vitamin A deficiency. A striking finding is the high riboflavin level in Karat, including high levels of uncharacterized flavonoids. Niacin and α-tocopherol are at levels that may contribute importantly to dietary intake within normal patterns of consumption. These data present a more complete basis for promoting the nutritional benefits of these banana cultivars where they are consumed in the Pacific, and potential benefits for promoting elsewhere.

Carotenoid content and flesh color of selected banana cultivars growing in Australia.

Background: The problems of vitamin A deficiency and chronic diseases have emerged in recent years in some countries in the Micronesian region. These problems are associated with the dietary shift towards imported processed foods and lifestyle changes. Research in the Federated States of Micronesia indicates that yellow- and orange-fleshed banana cultivars contain significant levels of provitamin A carotenoids. Objective: To identify further banana cultivars that may be promoted to alleviate vitamin A deficiency among children and women and chronic disease problems among adults. Methods: Ripe fruit of banana cultivars growing in Australia (sourced mostly from a field research collection) were assessed for carotenoid content and flesh color. Ten cultivars with yellow or yellow/orange flesh color (including common cultivars of Southeast Asia and the Pacific Islands) were selected and compared with two cream-fleshed cultivars, including Williams, of the Cavendish group, the most commonly marketed banana worldwide. Carotenoid content was analyzed by high-performance liquid chromatography (HPLC). Flesh color was analyzed by HunterLab colorimetry. Results: The yellow/orange-fleshed Asupina (a Fe'i banana) contained the highest level (1,412 μg/100 g) of trans β-carotene, the most important provitamin A carotenoid, a level more than 20 times higher than that of Williams. All 10 yellow or yellow/orange-fleshed cultivars (Asupina, Kirkirnan, Pisang Raja, Horn Plantain, Pacific Plantain, Kluai Khai Bonng, Wain, Red Dacca, Lakatan, and Sucrier) had significant carotenoid levels, potentially meeting half or all of the estimated vitamin A requirements for a nonpregnant, nonlactating adult woman within normal consumption patterns. All were acceptable for taste and other attributes. The cream-fleshed cultivars had minimal carotenoid levels. There was a positive significant correlation between carotenoid content and deeper yellow/orange coloration indicators. Conclusions: These yellow- or yellow/orange-fleshed carotenoid-rich banana cultivars should be considered for promotion in order to alleviate vitamin A deficiency and chronic disease in susceptible target communities and to provide variety and enjoyment as exotic fruits in both developing and industrialized countries.

Productivity, carbon assimilation and intra-annual change in tropical reef platform seagrass communities of the Torres Strait, north-eastern Australia

Detailed data on seagrass distribution, abundance, growth rates and community structure information were collected at Orman Reefs in March 2004 to estimate the above-ground productivity and carbon assimilated by seagrass meadows. Seagrass meadows were re-examined in November 2004 for comparison at the seasonal extremes of seagrass abundance. Ten seagrass species were identified in the meadows on Orman Reefs. Extensive seagrass coverage was found in March (18,700 ha) and November (21,600 ha), with seagrass covering the majority of the intertidal reef-top areas and a large proportion of the subtidal areas examined. There were marked differences in seagrass above-ground biomass, distribution and species composition between the two surveys. Major changes between March and November included a substantial decline in biomass for intertidal meadows and an expansion in area of subtidal meadows. Changes were most likely a result of greater tidal exposure of intertidal meadows prior to November leading to desiccation and temperature-related stress. The Orman Reef seagrass meadows had a total above-ground productivity of 259.8 t DW day-1 and estimated carbon assimilation of 89.4 t C day-1 in March. The majority of this production came from the intertidal meadows which accounted for 81% of the total production. Intra-annual changes in seagrass species composition, shoot density and size of meadows measured in this study were likely to have a strong influence on the total above-ground production during the year. The net estimated above-ground productivity of Orman Reefs meadows in March 2004 (1.19 g C m-2 day-1) was high compared with other tropical seagrass areas that have been studied and also higher than many other marine, estuarine and terrestrial plant communities.

A comparison of a blocking ELISA and a haemagglutination inhibition assay for the detection of antibodies to Avibacterium (Haemophilus) paragallinarum in sera from artificially infected chickens

The ability of blocking ELISAs and haemagglutination-inhibition (HI) tests to detect antibodies in sera from chickens challenged with either Avibacterium (Haemophilus) paragallinarum isolate Hp8 (serovar A) or H668 (serovar C) was compared. Serum samples were examined weekly over the 9 weeks following infection. The results showed that the positive rate of serovar A specific antibody in the B-ELISA remained at 100% from the second week to the ninth week. In chickens given the serovar C challenge, the highest positive rate of serovar C specific antibody in the B-ELISA appeared at the seventh week (60% positive) and was then followed by a rapid decrease. The B-ELISA gave significantly more positives at weeks 2, 3, 7, 8 and 9 post-infection for serovar A and at week 7 post-infection for serovar C. In qualitative terms, for both serovar A and serovar C infections, the HI tests gave a lower percentage of positive sera at all time points except at 9 weeks post-infection with serovar C. The highest positive rate for serovar A HI antibodies was 70% of sera at the fourth and fifth weeks post-infection. The highest rate of serovar C HI antibodies was 20% at the fifth and sixth weeks post-infection. The results have provided further evidence of the suitability of the serovar A and C B-ELISAs for the diagnosis of infectious coryza.

An evaluation of the apxIVA based PCR-REA method for differentiation of Actinobacillus pleuropneumoniae

A restriction analysis of PCR (PCR-REA) amplified apxIVA gene has been suggested as an alternative method for serotyping of Actinobacillus pleuropneumoniae by Jaglic et al. [Jaglic, Z., Svastova, P., Rychlik, I., Nedbalcova, K., Kucerova, Z., Pavlik, I., Bartos, M., 2004. Differentiation of Actinobacillus pleuropneumoniae by PCR-REA based on sequence variability of the apxIVA gene and by ribotyping. Vet. Microbiol. 103, 63-69]. The current study investigated whether this alternative method could distinguish between the reference strains of serovars 13-15 and the value of the method when applied to 47 field isolates representing serovars 1-3, 5, 7-9, 12 and 15 as well as non-typable isolates. The reference strains of serovars 13 and 14 had the same sized product after the apxIVA PCR, while the product for serovar 15 was of different size compared to all the other serovar reference strains. The CfoI digest profiles of the reference serovars 13 and 14 strains were different from each other and from all other serovars. The HpaII digest profiles of these two serovars were very similar to each other, but both were distinctively different from the other serovar profiles. The CfoI digest profile of serovar 15 strain was very similar to the serovars 3 and 12 strains except for two faint extra bands for serovar 15. The HpaII digest profiles of serovars 12 and 15 reference strains were identical. The PCR-REA method correctly recognized the serovar of 21 of 43 field isolates. It was concluded that the method was a useful additional tool to support, but could not replace, conventional serotyping.

Spatial model of site index based on Y-ray spectrometry and a digital elevation model for two Pinus species in Tuan Toolara State Forest, Queensland, Australia

Site index prediction models are an important aid for forest management and planning activities. This paper introduces a multiple regression model for spatially mapping and comparing site indices for two Pinus species (Pinus elliottii Engelm. and Queensland hybrid, a P. elliottii x Pinus caribaea Morelet hybrid) based on independent variables derived from two major sources: g-ray spectrometry (potassium (K), thorium (Th), and uranium (U)) and a digital elevation model (elevation, slope, curvature, hillshade, flow accumulation, and distance to streams). In addition, interpolated rainfall was tested. Species were coded as a dichotomous dummy variable; interaction effects between species and the g-ray spectrometric and geomorphologic variables were considered. The model explained up to 60% of the variance of site index and the standard error of estimate was 1.9 m. Uranium, elevation, distance to streams, thorium, and flow accumulation significantly correlate to the spatial variation of the site index of both species, and hillshade, curvature, elevation and slope accounted for the extra variability of one species over the other. The predicted site indices varied between 20.0 and 27.3 m for P. elliottii, and between 23.1 and 33.1 m for Queensland hybrid; the advantage of Queensland hybrid over P. elliottii ranged from 1.8 to 6.8 m, with the mean at 4.0 m. This compartment-based prediction and comparison study provides not only an overview of forest productivity of the whole plantation area studied but also a management tool at compartment scale.

Health and medicinal benefits of persimmon fruit: A review

The anti-oxidant activity of persimmon fruit appears to be mainly due to its high-molecular-weight tannin content. Antioxidant activity is variety specific with some astringent varieties showing very high antioxidant activity, comparable to strawberry and blueberry. In vitro, and limited animal studies, have shown that condensed tannins in the fruit may reduce the risk of cardiovascular disease, hypertension, diabetes and a wide range of cancers. Persimmon has an unusual property in that it appears to alter and reduce the rate of alcohol absorption and metabolism and thus ameliorate the symptoms of a hangover. The health and medicinal benefits of persimmon are considerable and should be further researched and promoted by persimmon industries around the world.

Effect of DNA extraction on ageing success in coral trout (Plectropomus leopardus) otoliths

To maximize the information commonly collected from otoliths, the effect of DNA extraction on the estimation of age with otoliths was evaluated by comparing sagittal otolith samples from common coral trout (Plectropomus leopardus) for clarity and ageing discrepancies in DNA-extracted and untreated control otoliths. The DNA extraction process had no significant effect, indicating that archived otoliths can be used as a source of DNA while retaining their utility for age estimation.

Flour yield QTLs in three Australian doubled haploid wheat populations

Flour yield quantitative trait loci (QTLs) were identified in 3 Australian doubled haploid populations, Sunco × Tasman, CD87 × Katepwa, and Cranbrook × Halberd. Trial data from 3 to 4 sites or years were available for each population. QTLs were identified on chromosomes 2BS, 4B, 5AL, and 6BL in the Sunco × Tasman population, on chromosomes 4B, 5AS, and 6DL in the CD87 × Katepwa population, and on chromosomes 4DS, 5DS, and 7AS in the Cranbrook × Halberd population. In the Sunco × Tasman cross the highest genetic variance was detected with the QTL on chromosome 2B (31.3%), in the CD87 × Katepwa cross with the QTL on chromosome 4B (23.8%), and in the Cranbrook × Halberd cross with the QTL on chromosome 5D (18%). Only one QTL occurred in a similar location in more than one population, indicating the complexity of the flour yield character across different backgrounds.