Use of monoclonal antibodies in blocking ELISA detection of transmissible gastroenteritis virus in faeces of piglets

Monoclonal antibodies (mAb) to the transmissible gastroenteritis virus (TGEV) nucleoprotein (N) and membrane protein (M) were prepared and used for the comparative assessment of three blocking ELISA variants to detect TGEV. The competitive blocking ELISA format showed the highest sensitivity, allowing detection of 10(3) TCID50 TGEV/ml in culture medium. Ninety-nine porcine field faecal samples obtained from 37 herds affected with diarrhoea were examined, and various TGEV levels were found in nine samples from six herds. However, only in three samples were significant TGEV concentrations demonstrated. The relationship between incidence of TGEV gastroenteritis and the spread of porcine respiratory coronavirus infection in pig farms is discussed.     

Verification of sensitivity and specificity of group a rotavirus detection in piglets faeces with monoclonal blocking ELISA methods

Monoclonal antibodies to group A rotavirus Vp6 protein were prepared and used for verification of three blocking enzyme-linked immunosorbent assay (ELISA) modifications to detect rotavirus A. Selected competitive blocking ELISA (CB-ELISA) and electron microscopy (EM) were used for examination of 194 field faecal samples of piglets affected with diarrhoea. Rotavirus was detected in 43 samples (22.2%) by CB-ELISA method, whereas in 26 (13.4%) samples by EM examination. However, of 26 samples positive by EM, rotavirus A was detected by CB-ELISA in 19 (73.1%) samples; indicating the share of group A rotavirus in all cases of gastroenteritis caused by rotavirus. The sensitivity and specificity of the CB-ELISA was verified both by inclusion of control samples containing transmissible gastroenteritis virus (TGEV) and porcine epidemic diarrhoea virus (PEDV) in each analysis and by comparative examination of samples with the commercial ELISA kit. The CB-ELISA sensitivity was positively affected by examination of samples in the presence of chelating agent.     

Characterization of flavonol conjugates in immature leaves of pak choi [Brassica rapa L. Ssp. chinensis L. (Hanelt.)] by HPLC-DAD and LC-MS/MS

The flavonoid composition of immature leaves of pak choi [Brassica rapa L. ssp. chinensis L. (Hanelt.)] was investigated. Flavonol aglycone content was measured in 11 pak choi varieties, indicating significant differences (P < 0.05) in content between varieties and relatively high contents of kaempferol and isorhamnetin. Levels of quercetin ranged from 3.2 to 6.1 mg/100 g of dry weight (DW), whereas levels of isorhamnetin and kaempferol were significantly higher (8.1-35.1 and 36.0-102.6 mg/100 g of DW, respectively). A large number of glycoside and hydroxycinnamic acid derivatives of quercetin, kaempferol, and isorhamnetin were identified in cv. 'Shanghai' by LC/UV-DAD/ESI-MS/MS. The UV-DAD data allowed identification of hydroxycinnamic acid derivatives, but detailed MS/MS fragmentations were required for the structure elucidation. Pak choi could be a potentially important source of dietary flavonols, in particular, kaempferol and isorhamnetin.     

Morphological characterization of seeds of three Australian wild Citrus species (Rutaceae): Citrus australasica F. Muell., C. inodora F.M. Bailey and C. garrawayi F.M. Bailey

The comparative morphology of the seeds of three Australian Citrus species, C. australasica C. inodora and C. garrawayi, was studied. Their seed characteristics were broadly similar to those of the cultivated species of the genus, when observed under light and scanning electron microscopy. Citrus garrawayi differed in seed shape (rounded to triangular) and seed coat morphology (i.e., thicker with longer epidermal protrusions) from C. australasica and C. inodora (rounded surface with flat underside in shape). The well-developed minute epidermal protrusions on the seed coat of C. garrawayi were more similar to those in the cultivated species, C. × sinensis and C. × aurantium. In contrast, the surface topography of C. australasica and C. inodora seeds was more like that of the cultivated species, C. × aurantifolia and C. × limon. Seed morphology, especially surface topography, was found to be a useful tool for taxonomic identification in Australian wild citrus.     

Cold tolerance during juvenile development in sorghum: a comparative analysis by genomewide association and linkage mapping

Improved cold tolerance during the juvenile phase is a major breeding goal to develop new sorghum cultivars suitable as an alternative energy crop in temperate regions. The objectives of this study were to identify marker‐trait associations for cold tolerance in a sorghum diversity panel fingerprinted with 2620 single nucleotide polymorphism (SNP) markers and to detect quantitative trait loci (QTL) in two F_2:3 populations. Traits of interest were dry matter growth rate (DMGR), leaf appearance rate (LAR), chlorophyll content (SPAD) and chlorophyll fluorescence (F_v′/F_m′ and Ф_PSII) in relation to temperature. The association panel comprised 194 genotypes, while the F_2:3 populations consisted of 80 and 92 genotypes. All populations were tested under controlled conditions in a minimum of four temperature regimes ranging from 9.4°C to 20.8°C. QTL were identified for means across environments and regression parameters describing temperature effects. Several marker‐trait associations for mean (m) DMGR, base temperature (T_b) of SPAD and Ф_PSII and temperature effect on LAR were validated by QTL detected in population 1 or 2. Promising QTL regions were found on chromosomes SBI‐01, SBI‐02, SBI‐03, SBI‐04, SBI‐06 and SBI‐09, among them genomic regions where candidate genes involved in the C‐repeat binding pathway or encoding cold‐shock proteins are located.     

Assessing and managing data‐limited ornamental fisheries in coral reefs

Coral reefs support numerous ornamental fisheries, but there are concerns about stock sustainability due to the volume of animals caught. Such impacts are difficult to quantify and manage because fishery data are often lacking. Here, we suggest a framework that integrates several data‐poor assessment and management methods in order to provide management guidance for fisheries that differ widely in the kinds and amounts of data available. First, a resource manager could assess the status of the ecosystem (using quantitative metrics where data are available and semi‐quantitative risk assessment where they are not) and determine whether overall fishing mortality should be reduced. Next, productivity susceptibility analysis can be used to estimate vulnerability to fishing using basic information on life history and the nature of the fishery. Information on the relative degree of exploitation (e.g. export data or ratios of fish density inside and outside no‐take marine reserves) is then combined with the vulnerability ranks to prioritize species for precautionary management and further analysis. For example, species that are both highly exploited and vulnerable are good candidates for precautionary reductions in allowable capture. Species that appear to be less vulnerable could be managed on a stock‐specific basis to prevent over‐exploitation of some species resulting from the use of aggregate catch limits. The framework could be applied to coral reef ornamental fisheries which typically lack landings, catch‐per‐unit‐effort and age‐size data to generate management guidance to reduce overfishing risk. We illustrate the application of this framework to an ornamental fishery in Indonesia.     

Australian Oryza: Utility and Conservation

Australian Oryza are an understudied and underexploited genetic resource for rice improvement. Four species are indigenous: Oryza rufipogon, Oryza meridionalis, Oryza australiensis are widespread across northern Australia, whereas Oryza officinalis is known from two localities only. Molecular analysis of these wild populations is required to better define the distinctness of the taxa and the extent of any gene flow between them and rice. Limited collections of these wild populations are held in seed and DNA banks. These species have potential for domestication in some cases but also have many traits of potential value in the improvement of domesticated rice. Stress tolerance (biotic and abiotic) and grain quality characteristics in these populations may be useful.     

A randomized, double-blind, placebo-controlled study to evaluate the effect of EFF1001, an Actinidia arguta (hardy kiwi) preparation, on CADESI score and pruritus in dogs with mild to moderate atopic dermatitis

Canine atopic dermatitis (AD) is common and new therapies are beneficial. This multicentric, randomized, double-blind, placebo-controlled study tested the efficacy of Actinidia arguta (hardy kiwi) (EFF1001) in dogs with mild/moderate AD. The study was divided into two stages. Stage 1 lasted 6 weeks. In the first 2 weeks prednisolone [days 1–3: 0.2 mg/kg twice daily (BID), days 4–14: 0.2 mg/kg every other day (EOD)] was administered. Responsive dogs were placed on prednisolone 0.2 mg/kg EOD + assigned test article [either placebo or EFF1001 (30 mg/kg)] once daily for 4 weeks. Stage 1 responders were advanced to stage 2, which involved 4 weeks of just EFF1001. Clinicians scored lesions using Canine Atopic Dermatitis Extent and Severity Index (CADESI) and owners scored pruritus using a Pruritus Visual Analogue Scale. Seventy-seven dogs were enrolled, 76 were randomized on day 14, and 57 (57/76 = 75%) completed stage 1 (27 in EFF1001 and 30 in placebo). At the end of stage 1, 35 of 57 dogs (35/57 = 61%) responded (18 in EFF1001 and 17 in placebo) and advanced to stage 2. At completion of stage 1, CADESI scores did not significantly differ between groups while pruritus decreased in EFF1001 group and approached significance. At completion of stage 2, 19 dogs (19/35 = 54%) responded (15/19 = 79% had received EFF1001 and 4/19 = 21% placebo in stage 1). After completing stage 2, dogs placed on EFF1001 throughout the study were 3.5 times more likely to either maintain or improve scores than those that started it in stage 2. It is concluded that EFF1001 is beneficial adjunctive therapy after prolonged use.