Using chemical and DNA marker analysis to authenticate a high-value food, manuka honey.

Ensuring the authenticity of food is a rapidly emerging issue, especially in regard to high-value products that are marketed through increasingly complex global food chains. With the ever-increasing potential for mislabeling, fraud and adulteration, governments are increasingly having to invest in, and assure, the authenticity of foods in international trade. This is particularly the case for manuka honey, an iconic New Zealand food product. We show how the authenticity of a specific type of honey can be determined using a combination of chemicals derived from nectar and DNA derived from pollen. We employ an inter-disciplinary approach to evaluate a selection of authenticity markers, followed by classification modelling to produce criteria that consistently identify manuka honey from New Zealand. The outcome of our work provides robust identification criteria that can be applied in a regulatory setting to authenticate a high-value natural food. Our approach can transfer to other foods where assurance of authenticity must take into account a high level of natural variability.

Application of a pathogenicity marker found in Escherichia coli for the assessment of irrigation water quality.

A polymerase chain reaction (PCR)-based method was developed to differentiate between pathogenic and nonpathogenic Escherichia coli (E. coli). A pathogenicity marker, linked to the deletion of the ygfB gene, was identified in 80% of the clinical E. coli isolates tested. This marker, combined with the malic acid dehydrogenase gene, formed the duplex PCR that was subsequently used to screen E. coli isolates recovered from two secondary wastewater treatment plants (STPs) and a river site. All waters samples are used to irrigate dairy farm pasture in the West Gippsland region of Victoria, Australia. Results from three consecutive months of sampling (December 2001 and January and February 2002) indicated that Longwarry STP showed 8, 8, and 0% pathogenic E. coli; Pakenham STP showed 0, 12.5, and 33%; and the Bunyip river site showed 20, 12, and 25% respectively.

Design of a detection survey for Ostreid herpesvirus-1 using hydrodynamic dispersion models to determine epidemiological units.

Using Ostreid herpesvirus-1 (OsHV-1) as a case study, this paper considers a survey design methodology for an aquatic animal pathogen that incorporates the concept of biologically independent epidemiological units. Hydrodynamically-modelled epidemiological units are used to divide marine areas into sensible sampling units for detection surveys of waterborne diseases. In the aquatic environment it is difficult to manage disease at the animal level, hence management practices are often aimed at a group of animals sharing a similar risk. Using epidemiological units is a way to define these groups, based on a similar level of probability of exposure based on the modelled potential spread of a viral particle via coastal currents, that can help inform management decisions.

Optimization of Novel Indazoles as Highly Potent and Selective Inhibitors of Phosphoinositide 3-Kinase δ for the Treatment of Respiratory Disease.

Optimization of lead compound 1, through extensive use of structure-based design and a focus on PI3Kδ potency, isoform selectivity, and inhaled PK properties, led to the discovery of clinical candidates 2 (GSK2269557) and 3 (GSK2292767) for the treatment of respiratory indications via inhalation. Compounds 2 and 3 are both highly selective for PI3Kδ over the closely related isoforms and are active in a disease relevant brown Norway rat acute OVA model of Th2-driven lung inflammation.

Barcoding and border biosecurity: identifying cyprinid fishes in the aquarium trade.

BACKGROUND: Poorly regulated international trade in ornamental fishes poses risks to both biodiversity and economic activity via invasive alien species and exotic pathogens. Border security officials need robust tools to confirm identifications, often requiring hard-to-obtain taxonomic literature and expertise. DNA barcoding offers a potentially attractive tool for quarantine inspection, but has yet to be scrutinised for aquarium fishes. Here, we present a barcoding approach for ornamental cyprinid fishes by: (1) expanding current barcode reference libraries; (2) assessing barcode congruence with morphological identifications under numerous scenarios (e.g. inclusion of GenBank data, presence of singleton species, choice of analytical method); and (3) providing supplementary information to identify difficult species. METHODOLOGY/PRINCIPAL FINDINGS: We sampled 172 ornamental cyprinid fish species from the international trade, and provide data for 91 species currently unrepresented in reference libraries (GenBank/Bold). DNA barcodes were found to be highly congruent with our morphological assignments, achieving success rates of 90-99%, depending on the method used (neighbour-joining monophyly, bootstrap, nearest neighbour, GMYC, percent threshold). Inclusion of data from GenBank (additional 157 spp.) resulted in a more comprehensive library, but at a cost to success rate due to the increased number of singleton species. In addition to DNA barcodes, our study also provides supporting data in the form of specimen images, morphological characters, taxonomic bibliography, preserved vouchers, and nuclear rhodopsin sequences. Using this nuclear rhodopsin data we also uncovered evidence of interspecific hybridisation, and highlighted unrecognised diversity within popular aquarium species, including the endangered Indian barb Puntius denisonii. CONCLUSIONS/SIGNIFICANCE: We demonstrate that DNA barcoding provides a highly effective biosecurity tool for rapidly identifying ornamental fishes. In cases where DNA barcodes are unable to offer an identification, we improve on previous studies by consolidating supplementary information from multiple data sources, and empower biosecurity agencies to confidently identify high-risk fishes in the aquarium trade.