PP2A Inhibition Assay Using Recombinant Enzyme for Rapid Detection of Okadaic Acid and Its Analogs in Shellfish

Okadaic acid and its analogs (OAs) responsible for diarrhetic shellfish poisoning (DSP) strongly inhibit protein phosphatase 2A (PP2A) and thus are quantifiable by measuring the extent of the enzyme inhibition. In this study, we evaluated the suitability of the catalytic subunit of recombinant human PP2A (rhPP2Ac) for use in a microplate OA assay. OA, dinophysistoxin-1(DTX1), and hydrolyzate of 7-O-palmitoyl-OA strongly inhibited rhPP2Ac activity with IC50 values of 0.095, 0.104, and 0.135 nM, respectively. The limits of detection and quantitation for OA in the digestive gland of scallops and mussels were 0.0348 μg/g and 0.0611 μg/g respectively, and, when converted to the whole meat basis, are well below the regulation level proposed by EU (0.16 μg/g whole meat). A good correlation with LC-MS data was demonstrated, the correlation coefficient being 0.996 with the regression slope of 1.097

eDNA metabarcoding analysis reveals the consequence of creating ecosystem‐scale refugia from deer grazing for the soil microbial communities

シカの森林被害は土壌微生物にも波及する --大規模生態系操作実験と環境DNA分析の融合--. 京都大学プレスリリース. 2023-12-22.Ungulate overbrowsing is a growing problem in forests worldwide due to its prolonged and pervasive impact on plant biodiversity and ecosystem functioning. It has been shown that overbrowsing not only reduces plant species diversity and biomass (i.e., direct effects) but also causes a loss of associated trophic levels that could potentially feedback to influence plant community structure (i.e., indirect effects). One of the primary pathways of such indirect effects that have not been fully examined is the impact of overbrowsing on soil microorganisms. Recent studies have shown that soil microorganisms maintain vegetation diversity and drive succession, so it is of critical importance to understand how soil microbial communities might be affected by or protected from the deer impact. To assess the consequence of creating artificial grazing refugia on the structure and composition of soil microbial communities, we compared the distribution and abundance of soil microbial taxa (bacteria, archaea, fungi) at the fenced versus unfenced control sites in the context of a catchment-scale field experiment in Japan. The eDNA metabarcoding analysis of soil microbial communities showed that the numbers of archaea and basidiomycetes fungal species were greater in the fenced site than in the control, while no such pattern was found for bacteria and ascomycetes fungi. Despite the lack of significant influence of the fence treatment on taxonomic composition in the soil fungal communities, their functional guild composition was influenced by the fenced treatment, with significant changes in the abundance of animal pathogens. Thus, although the effect of fencing on soil microbial communities is characterized by complex responses that vary from taxon to taxon, our work suggests that creating ecosystem-scale refugia from deer overgrazing might help sustain certain, if not all, taxa of soil microbial communities

Feeding effect of selenium enriched rotifers on larval growth and development in red sea bream Pagrus major

Feeding trials were conducted to investigate the effect of selenium (Se)-enriched rotifers on growth and development of red sea bream Pagrus major larvae. Fish were reared from fertilized eggs (98% hatch rate) to 20. days post hatch (dph) at 19. °C with two different food sources; non-enriched S-type rotifers (0.0. μg. Se/g D.W., control diet) or Se-enriched rotifers (2.2. μg. Se/g D.W., Se-enriched diet) at 10. rotifers/mL, respectively. On the last day of larviculture, the Se-enriched diet accelerated growth and developmental stage of fish larvae. The larvae fed Se-enriched rotifers were advanced in the following parameters compared to those fed control diet: total length (6.06 vs 5.53. mm), standard length (5.74 vs 5.26. mm), head length (1.46 vs 1.28. mm), eye diameter (0.57 vs 0.50. mm), the number of caudal fin rays (5.8 vs 1.9), and the proportion of individuals undergoing notochord flexion (55 vs 3%). Fish larvae of 20. dph showed higher Se concentration (9.5 ± 0.2. μg/g DW) with the Se-enriched diet than with the control diet (1.3 ± 0.3. μg/g DW), but there were no significant differences in the composition of polyunsaturated fatty acids which significantly affect larval growth and development. Therefore, the feeding of Se enriched rotifers enhanced growth and development of the red sea bream P. major larvae