Detection of SNPs in fish DNA: application of the fluorogenic ribonuclease protection (FRIP) assay for the authentication of food contents

The fluorogenic ribonuclease protection (FRIP) assay was used to detect single nucleotide polymorphisms (SNPs) in commercially produced fish products. By using fluorescence resonance energy transfer (FRET) between fluorophore and quencher labeled probes, the species-specific cleavage of sample RNA was detected by measuring the fluorescence intensity during the FRIP assay. We were able to discriminate raw and thermally processed eel and tuna species using the FRIP-based SNP detection method. Furthermore, the intensity of fluorescence was correlated with the mutant/wild-type ratio. These results suggest that the FRIP assay is a useful method for the in situ confirmation of labels of fishery foods during food production.     

Collembolan community dynamics during deciduous forests regeneration in Japan

The structure and feeding group composition of collembolan communities were studied in secondary deciduous forests of different ages to investigate the collembolan community response to environmental changes associated with forest cycles. The study was carried out at eight sites forming a chronosequence (1, 4, 12, 24, 51, 54, 71 and 128 years after clear cutting) of deciduous forest stands in northern Ibaraki (Japan). Total collembolan density and species richness was low at the 1-year-old site, and there was little difference in density among sites over 4 years of age. The density of sucking feeders was especially low at the 1-year-old site. Species richness of trees of a diameter at breast height (DBH)<5 cm positively correlated with the density of fungal feeders. Species richness of total Collembolans and of sucking feeders correlated positively with the water content of the organic layer. Ordination of the collembolan community with Canonical Correspondence Analysis suggested that species richness of larger trees (DBH 5 cm) contributed to the differences in species composition of fungal feeders and sucking feeders. We conclude that total abundance and species richness of collembolans recovered within 4 years after clear-cutting, but species composition of fungal feeders and sucking feeders took longer to recover.     

Genotyping of the <Emphasis Type="Italic">SiDREB2</Emphasis> gene in worldwide foxtail millet (<Emphasis Type="Italic">Setaria italica</Emphasis> (L.) P. Beauv.) genetic resources with special attention to Indian landraces

We genotyped Setaria italica Dehydration-responsive element-binding 2 (SiDREB2) gene, which had been reported to be associated with dehydration stress tolerance, in 588 accessions of foxtail millet from various parts of Eurasia and other regions by a dCAPS marker. Of these, 480 accessions were genotyped in ribosomal DNA (rDNA), polyphenol oxidase (PPO) gene and Heading date 1 (HD1) gene in our previous studies, and 108 accessions from India were newly used in this study and genotyped in these genes in addition to the SiDREB2 gene. We compared the geographical distribution of the SiDREB2 genotypes with that of these three genes. Accessions from countries in South Asia including India and Sri Lanka were the most variable in the SiDREB2 gene followed by Korean accessions and Japanese accessions, suggesting that Indian accessions and East Asian accessions are useful as genetic resources for dehydration stress tolerance. This study also showed that although Indian accessions are not so diverse in rDNA and in transposon display markers previously studied, they are diverse in adaptive genes such as SiDREB2, PPO and HD1 genes.     

黑龙江省主栽水稻品种抽穗模型研究初报

采用黑龙江省生产上主栽的早熟、中熟和晚熟的6个水稻品种,在哈尔滨地区做分期栽培田间试验。调查记载播种、插秧和抽穗期。采取平行观测的气象资料,分析温度对水稻抽穗期和水稻营养生长期的影响。结果表明:不同的插秧期水稻抽穗期变化很大,不同熟期的水稻品种插秧至抽穗期需要的有效积温差别明显,但每个品种需要的有效积温很稳定。     

Diagnostic value of recombinant nanoluciferase fused Toxoplasma gondii antigens in Luciferase-linked Antibody Capture Assay (LACA) for Toxoplasma infection in pigs

Toxoplasmosis is a widespread protozoan zoonosis. Since ingesting undercooked meat harboring Toxoplasma gondii cyst is considered one of the major transmission routes to humans, the screening of T. gondii in meat-producing animals can reduce the risk of food-borne toxoplasmosis in humans. Among serological diagnostic methods, Luciferase-linked Antibody Capture Assay (LACA) has been found to be a promising platform with high sensitivity and specificity. In this study, we aimed to evaluate recombinant nanoluciferase fused-T. gondii antigens (rNluc-GRA6, rNluc-GRA7, rNluc-GRA8 and rNluc-BAG1) for their potential use in LACA for pigs. As a result, the sensitivity of GRA6-, GRA7-, GRA8- and BAG1-LACA were 70.0%, 80.0%, 80.0% and 30.0% with specificity 87.0%, 81.5%, 74.1% and 50.0%, respectively. The cocktail LACA using a mixture of rNluc-GRA6, rNluc-GRA7 and rNluc-GRA8 indicated higher sensitivity (90.0%) and a similar specificity (96.3%) in comparison with the commercial ELISA kit. Compared to the Dye-Test as a reference test, cocktail LACA showed strong agreement (kappa value=0.811) when we assessed pig sera collected at the slaughterhouse. In addition, we also successfully established the rapid LACA format for the detection of Toxoplasma infection in pigs (called Rapid-LACA) in which the test could be performed within 30 min. In Rapid-LACA, the protein A pre-coated/blocked plates could be preserved at −30°C, 4°C or room temperature conditions for at least two months without compromising on the quality of assay.