Evaluation of housekeeping genes as references for quantitative real-time PCR analysis of gene expression in the murrel <Emphasis Type="Italic">Channa striatus</Emphasis> under high-temperature stress

Quantitative real-time polymerase chain reaction is the most advanced method of quantifying gene expression studies; however, the significance of the obtained results strongly depends on the normalization of the data to compensate for differences between the samples. In the present study, expression analysis of six different constitutively expressed genes viz. 18S ribosomal RNA, glyceraldehyde-3-phosphate dehydrogenase (gapdh), beta actin (βactin), ribosomal binding protein L13, tubulin and TATA-box-binding protein (tbp) were carried out to test their efficacy as reference genes in three different tissues, namely liver, gill and muscle of murrel Channa striatus exposed to high temperature for variable time periods. The stability and suitability of the genes were determined by using bioinformatic tools: GeNorm, NormFinder and BestKeeper. Based on the results, tub/βactin could be used as the reference genes for liver and gill tissues and βactin/gapdh could be the reference genes for muscle tissues in Channa striatus under both short- and long-term thermal stress.     

Between ‘state’ and ‘society’: Commune authorities and the environment in Vietnam's craft villages

Maintaining a liveable environment in Vietnam's polluted craft villages is a daily challenge for state authorities and residents. Neighbouring urban populations demand that the state effectively curtails and manages pollution, while local residents prioritise their livelihoods and routinely flout regulations. The commune official, tasked with the seemingly impossible task of environmental regulation, occupies a fraught position, torn between the imperatives and constraints of craft producers and state regulatory demands. This study of water pollution in northern Vietnam's craft villages finds that commune officials' conflicted role in environmental governance is a central factor in the failure of the current environmental governance regime, and reflects the internally conflicted nature of the Vietnamese state.     

Dietary supplementation of curcumin augments heat stress tolerance through upregulation of <Emphasis Type="Italic">nrf-2</Emphasis>-mediated antioxidative enzymes and <Emphasis Type="Italic">hsps</Emphasis> in <Emphasis Type="Italic">Puntius sophore</Emphasis>

Heat stress is one of the major environmental concerns in global warming regime and rising temperature has resulted in mass mortalities of animals including fishes. Therefore, strategies for high temperature stress tolerance and ameliorating the effects of heat stress are being looked for. In an earlier study, we reported that Nrf-2 (nuclear factor E2-related factor 2) mediated upregulation of antioxidative enzymes and heat shock proteins (Hsps) provide survivability to fish under heat stress. In this study, we have evaluated the ameliorative potential of dietary curcumin, a potential Nrf-2 inducer in heat stressed cyprinid Puntius sophore. Fishes were fed with diet supplemented with 0.5, 1.0, and 1.5% curcumin at the rate 2% of body weight daily in three separate groups (n = 40 in each group) for 60 days. Fishes fed with basal diet (without curcumin) served as the control (n = 40). Critical thermal maxima (CTmax) was determined for all the groups (n = 10, in duplicates) after the feeding trial. Significant increase in the CTmax was observed in the group fed with 1.5% curcumin- supplemented fishes whereas it remained similar in groups fed with 0.5%, and 1% curcumin-supplemented diet, as compared to control. To understand the molecular mechanism of elevated thermotolerance in the 1.5% curcumin supplemented group, fishes were given a sub-lethal heat shock treatment (36 °C) for 6 h and expression analysis of nrf-2, keap-1, sod, catalase, gpx, and hsp27, hsp60, hsp70, hsp90, and hsp110 was carried out using RT-PCR. In the gill, expression of nrf-2, sod, catalase, gpx, and hsp60, hsp70, hsp90, and hsp110 was found to be elevated in the 1.5% curcumin-fed heat-shocked group compared to control and the basal diet-fed, heat-shocked fishes. Similarly, in the liver, upregulation in expression of nrf-2, sod, catalase, and hsp70 and hsp110 was observed in 1.5% curcumin supplemented and heat shocked group. Thus, this study showed that supplementation of curcumin augments tolerance to high temperature stress in P. sophore that could be attributed to nrf-2-induced upregulation of antioxidative enzymes sod, catalase, gpx, and the hsps.     

Biomimetic Acid Soil Amelioration using Indigenous Soil Isolates in Comparison to other Potential Microorganism

In the present study, four different soil isolates were screened and evaluated for their acid neutralization capacity in liquid culture and soil column tests with either artificial soil or real acid soil. Miniaturized artificial soil column tests using Aspergillus fumigatus, Aureobasidium pullulans, and Myxococcus xanthus were also included. In the liquid-phase experiment, two isolates (designated ‘A’ and ‘B’) effectively increased the medium pH from 4 to 6.74 and 6.53, respectively, during the 6 d, and the ammonium-nitrogen (NH₄⁺-N) concentration increased from 400 ppm to 1000 ppm (isolate A). The isolates A and B were identified as Enterococcus hirae and Clostridium sp. in 16S ribosomal ribonucleic acid (rRNA) analysis. Artificial soil pH increased more quickly with soil isolates than in comparison to A. fumigatus or A. pullulans. In real acid soil pH increased from 4.2 to about pH 6.5 within 4 d. This suggests that biomimetic approaches could be valuable for acid soil amelioration.