Molecular discrimination of Perna (Mollusca: Bivalvia) species using the polymerase chain reaction and species-specific mitochondrial primers

This work was prompted by the need to be able to identify the invasive mussel species, Perna viridis, in tropical Australian seas using techniques that do not rely solely on morphology. DNA-based molecular methods utilizing a polymerase chain reaction (PCR) approach were developed to distinguish unambiguously between the three species in the genus Perna. Target regions were portions of two mitochondrial genes, cox1 and nad4, and the intergenic spacer between these that occurs in at least two Perna species. Based on interspecific sequence comparisons of the nad4 gene, a conserved primer has been designed that can act as a forward primer in PCRs for any Perna species. Four reverse primers have also been designed, based on nad4 and intergenic spacer sequences, which yield species-specific products of different lengths when paired with the conserved forward primer. A further pair of primers has been designed that will amplify part of the cox1 gene of any Perna species, and possibly other molluscs, as a positive control to demonstrate that the PCR is working.

Glucosinolate composition and anti-cancer potential of daikon and radish sprouts

Daikon and radish sprouts contain high levels of glucoraphenin, a glucosinolate which hydrolyses to form sulphoraphene. Sulphoraphene, like sulphoraphane from broccoli, is a potent inducer of phase 2 detoxification enzymes and consequently has potential anti-cancer action. Unlike broccoli however, daikon and radish do not possess epithiospecifier protein, a protein that inhibits conversion of glucosinolates to isothiocyanates, and consequently they may represent more suitable sources of phyto-chemicals with anti-cancer potential. Concentrations of glucoraphenin were highest in the seed, declining exponentially with sprout development. The rate of decline was observed to vary considerably between varieties of daikon and radish, with some varieties maintaining significantly high levels of glucoraphenin. Varieties maintaining a high level of glucoraphenin included 'Cherry Belle' and 'French Breakfast'.

Detection of equine herpesvirus type 1 using a real-time polymerase chain reaction.

Equid herpesvirus 1 (EHV1) is a major disease of equids worldwide causing considerable losses to the horse industry. A variety of techniques, including PCR have been used to diagnose EHV1. Some of these PCRs were used in combination with other techniques such as restriction enzyme analysis (REA) or hybridisation, making them cumbersome for routine diagnostic testing and increasing the chances of cross-contamination. Furthermore, they involve the use of suspected carcinogens such as ethidium bromide and ultraviolet light. In this paper, we describe a real-time PCR, which uses minor groove-binding probe (MGB) technology for the diagnosis of EHV1. This technique does not require post-PCR manipulations thereby reducing the risk of cross-contamination. Most importantly, the technique is specific; it was able to differentiate EHV1 from the closely related member of the Alphaherpesvirinae, equid herpesvirus 4 (EHV4). It was not reactive with common opportunistic pathogens such as Escherichia coli, Klebsiella oxytoca, Pseudomonas aeruginosa and Enterobacter agglomerans often involved in abortion. Similarly, it did not react with equine pathogens such as Streptococcus equi, Streptococcus equisimilis, Streptococcus zooepidemicus, Taylorella equigenitalis and Rhodococcus equi, which also cause abortion. The results obtained with this technique agreed with results from published PCR methods. The assay was sensitive enough to detect EHV1 sequences in paraffin-embedded tissues and clinical samples. When compared to virus isolation, the test was more sensitive. This test will be useful for the routine diagnosis of EHV1 based on its specificity, sensitivity, ease of performance and rapidity.

A field investigation of solubility and food chain accumulation of biosolid-cadmium across diverse soil types

One of the pathways for transfer of cadmium (Cd) through the food chain is addition of urban wastewater solids (biosolids) to soil, and many countries have restrictions on biosolid use to minimize crop Cd contamination. The basis of these restrictions often lies in laboratory or glasshouse experimentation of soil-plant transfer of Cd, but these studies are confounded by artefacts from growing crops in controlled laboratory conditions. This study examined soil to plant (wheat grain) transfer of Cd under a wide range of field environments under typical agronomic conditions, and compared the solubility and bioavailability of Cd in biosolids to soluble Cd salts. Solubility of biosolid Cd (measured by examining Cd partitioning between soil and soil solution) was found to be equal to or greater than that of soluble Cd salts, possibly due to competing ions added with the biosolids. Conversely, bioavailability of Cd to wheat and transfer to grain was less than that of soluble Cd salts, possibly due to addition of Zn with the biosolids, causing reduced plant uptake or grain loading, or due to complexation of soluble Cd2+ by dissolved organic matter.

Using a supply chain approach to guide R&D

Consumers today are presented with an increasing array of products. The growing competition for consumer expenditure requires a whole of supply chain approach to maintain market share for existing cultivars and to successfully commercialise new cultivars. The supply chain needs to deliver value and satisfaction to the end customer and profitability to their members. Critical to getting the product right is developing inherent robustness into the cultivar, and developing processes and systems through the whole supply chain that maintain product quality and add value. This paper describes the approach we have used in working with supply chains in Australia and Indonesia to identify priority areas for improvement. Our experience demonstrates the need for a champion in the supply chain with significant influence and a desire to improve. The paper also describes our approach towards improving a specific supply chain to achieve successful commercialisation of a new cultivar. The cultivar was primarily selected for good production characteristics and attractive visual appeal. The performance of the fruit is being monitored from farm to retail shelf to identify points where quality is lost and practices can be improved. A targeted R&D program is investigating ways of improving production efficiency (nutrition, flowering and canopy management), maturity standards to optimise flavour, harvesting and packing practices to reduce skin damage, and ripening and handling practices to optimise shelf life. This integrated approach is based on similar approaches used to improve the performance of existing mango and avocado cultivars.

Transmission of Japanese Encephalitis Virus from the Black Flying Fox, Pteropus alecto, to Culex annulirostris Mosquitoes, Despite the Absence of Detectable Viremia.

To determine the potential role of flying foxes in transmission cycles of Japanese encephalitis virus (JEV) in Australia, we exposed Pteropus alecto (Megachiroptera: Pteropididae) to JEV via infected Culex annulirostris mosquitoes or inoculation. No flying foxes developed symptoms consistent with JEV infection. Anti-JEV IgG antibodies developed in 6/10 flying foxes exposed to infected Cx. annulirostris and in 5/5 inoculated flying foxes. Low-level viremia was detected by real-time reverse transcriptase polymerase chain reaction in 1/5 inoculated flying foxes and this animal was able to infect recipient mosquitoes. Although viremia was not detected in any of the 10 flying foxes that were exposed to JEV by mosquito bite, two animals infected recipient mosquitoes. Likewise, an inoculated flying fox without detectable viremia infected recipient mosquitoes. Although infection rates in recipient mosquitoes were low, the high population densities in roosting camps, coupled with migratory behavior indicate that flying foxes could play a role in the dispersal of JEV.

Radish sprouts versus broccoli sprouts: a comparison of anti-cancer potential based on glucosinolate breakdown products.

Radish sprouts and broccoli sprouts have been implicated in having a potential chemoprotective effect against certain types of cancer. Each contains a glucosinolate that can be broken down to an isothiocyanate capable of inducing chemoprotective factors known as phase 2 enzymes. In the case of broccoli, the glucosinolate, glucoraphanin, is converted to an isothiocyanate, sulforaphane, while in radish a similar glucosinolate, glucoraphenin, is broken down to form the isothiocyanate, sulforaphene. When sprouts are consumed fresh (uncooked), however, the principal degradation product of broccoli is not the isothiocyanate sulforaphane, but a nitrile, a compound with little anti-cancer potential. By contrast, radish sprouts produce largely the anti-cancer isothiocyanate, sulforaphene. The reason for this difference is likely to be due to the presence in broccoli (and absence in radish) of the enzyme cofactor, epithiospecifier protein (ESP). In vitro induction of the phase 2 enzyme, quinone reductase (QR), was significantly greater for radish sprouts than broccoli sprouts when extracts were self-hydrolysed. By contrast, boiled radish sprout extracts (deactivating ESP) to which myrosinase was subsequently added, induced similar QR activity to broccoli sprouts. The implication is that radish sprouts have potentially greater chemoprotective action against carcinogens than broccoli sprouts when hydrolysed under conditions similar to that during human consumption.

Develop added value uses of high anti-bacterial honey

Honey is valued for its medicinal properties. There interest from industry to develop a rapid method to detect the bioactivity of honey and also to develop value added products from high antibacterial honey.

Increasing vegetable consumption: a means-end chain approach

Purpose – The purpose of this study is to illustrate how means-end chain theory can inform communications that effectively convey the health messages of vegetable consumption to various publics. Design/methodology/approach – Laddering interviews were conducted with 61 participants who consumed at least two serves of vegetables a day and were responsible in part or whole for shopping in their household. A means-end chain value map was then constructed using mecanalyst software. Findings – Using means-end theory, an example communications strategy was developed from the dominant chain. The health and wellness features that respondents associated with vegetables were “freshness”, a “source of vitamins and minerals”, and “high nutritional value”. In the mind of the consumer, these features were linked to the benefit concept “maintain energy and vitality”, which in turn was connected to the consequence “maintain an active life”. The end-states or goals participants ultimately connected to the health and wellness features of vegetables were that of “enjoy life” and “achieve goals”. Research limitations/implications – The research is limited in so far as subjects who consume less than two serves of vegetables are not recruited for this study. Practical implications – It is suggested that social marketing initiatives designed to increase vegetable consumption may base messages on health-related values or end-states of being to resonate more effectively with consumers. Social implications – High vegetable consumption is associated with a reduced risk of chronic disease. Effective strategies designed to increase vegetable consumption amongst populations may reduce the burden on health systems. Originality/value – This study illustrates how consumers' cognitive processes can inform social marketing communications.

ACIAR 9995 Improving the value chain for plantation-grown sawn wood

To improve the yield of marketable high value sawn and/or veneer timber from young plantation grown eucalypts: through the greater understanding and management of growth stresses; grain angle; juvenile wood; and other important wood properties.

Integrated pest management and supply chain improvement for mangoes in the Philippines and Australia

This project will incorporate two strategies for improving industry sustainability in the Philippines and Australia: (a) developing improved field management and quarantine monitoring and detection of mango pests and diseases; and (b) working with selected mango supply chains to identify and test areas for improvement.

Value chain technical development

Short-term research projects on insect pest management in processed food.

Detection and differentiation of bovine herpesvirus 1 and 5 using a multiplex real-time polymerase chain reaction.

A multiplex real-time PCR was developed for the detection and differentiation of two closely related bovine herpesviruses 1 (BoHV-1) and 5 (BoHV-5). The multiplex real-time PCR combines a duplex real-time PCR that targets the DNA polymerase gene of BoHV-1 and BoHV-5 and a real-time PCR targeting mitochondrial DNA, as a house-keeping gene, described previously by Cawthraw et al. (2009). The assay correctly identified 22 BoHV-1 and six BoHV-5 isolates from the Biosecurity Sciences Laboratory virus collection. BoHV-1 and BoHV-5 were also correctly identified when incorporated in spiked semen and brain tissue samples. The detection limits of the duplex assay were 10 copies of BoHV-1 and 45 copies of BoHV-5. The multiplex real-time PCR had reaction efficiencies of 1.04 for BoHV-1 and 1.08 for BoHV-5. Standard curves relating Ct value to template copy number had correlation coefficients of 0.989 for BoHV-1 and 0.978 for BoHV-5. The assay specificity was demonstrated by testing bacterial and viral DNA from pathogens commonly isolated from bovine respiratory and reproductive tracts. The validated multiplex real-time PCR was used to detect and differentiate BoHV-1 and BoHV-5 in bovine clinical samples with known histories.

Vegetables containing phytochemicals with potential anti-obesity properties: A review

The incidence of obesity is rising worldwide at an alarming rate and is becoming a major public health concern with incalculable social and economic costs. Studies have exposed the relationship between the adiposity, inflammation and the development of other metabolic disorders, so dietary factors that influence some or all of these are of interest. Dietary phytochemicals appear to be able to target different stages of the adipocyte (fat cell) lifecycle. For example, several classes of polyphenols have been implicated in suppressing the growth of adipose tissue through modifying the adipocyte lifecycle. Many dietary phytochemicals also have strong anti-inflammatory activity, but the amount present in plants varies and may be affected by processing. In this review we summarise the likely mechanisms of action of plant phytochemicals. We highlight the major vegetable sources of polyphenols, including those with possible synergistic attributes, discuss the variation in polyphenol levels and their distribution in cultivars and outline the effects of food processing. The identification and characterisation of the anti-obesogenic properties of phytochemicals in vegetables, as well as an appreciation of the effect of cooking on phytochemical content provide significant new information supporting dietary guidelines that encourage vegetable consumption for the prevention and management of lifestyle related disease.

Starchy Staples Value Chain Review

PARDI – Pacific Agribusiness Research & Development Initiative