miRNA and mRNA expression analysis reveals the effects of continuous heat stress on antibacterial responses to Aeromonas hydrophila lipopolysaccharide (LPS) in grass carp (Ctenopharyngodon idella).

Grass carp (Ctenopharyngodon idella) is the largest economic fish in freshwater culture in China, which is predisposed to infectious diseases under high temperature. Under the background of global warming, the industrialization of the Pearl River Delta region has led to aggravated thermal pollution, which has increasingly serious impacts on the aquatic ecological environment. This will result in more frequent exposure of grass carp to overheated water temperatures. Previous studies have only identified the regulatory genes of fish that respond to pathogens or temperature stress, but the transcriptional response to both is unknown. In this study, the histopathological analysis showed heat stress exacerbated spleen damage induced by Aeromonas hydrophila. The transcriptional responses of the spleens from A. hydrophila lipopolysaccharide (LPS) -injected grass carp undergoing heat stress and at normal temperatures for 6, 24, and 72 h were investigated by mRNA and microRNA sequencing. We identified 28, 20, and 141 differentially expressed (DE) miRNAs and 126, 383, and 4841 DE mRNAs between the two groups after 6, 24, and 72 h, respectively. There were 67 DE genes mainly involved in the cytochrome P450 pathway, antioxidant defense, inflammatory response, pathogen recognition pathway, antigen processing and presentation, and the ubiquitin-proteasome system. There were 5 DE miRNAs involved in regulating apoptosis and inflammation. We further verified 17 DE mRNAs and 5 DE miRNAs using quantitative real-time PCR. Based on miRNAs and mRNAs analysis, continuous heat stress will affect the antibacterial responses of grass carp spleens, resulting in aggravation of spleen injury. Together, these results provide data for further understanding of the decreased tolerance of fish to pathogen infection in persistent high-temperature environments.

Phosphate distribution and sources in the waters of Huangbai River, China: using oxygen isotope composition of phosphate as a tracer.

We investigated the distributions of phosphate (PO4-P) and used the oxygen isotope composition of phosphate (δ18OP) to quantify PO4-P sources in the waters of Huangbai River. According to the environmental characteristics of Huangbai River basin, the sampling stations in the Huangbai River were divided into three groups: sampling stations in the phosphate mining area, in the outcrop area of phosphate rock, and in the residential/agricultural area. The average PO4-P concentration was highest (2.34 ± 1.00 µmol/L) in the outcrop area of phosphate ore, intermediate in the residential/agricultural area (1.06 ± 1.21 µmol/L), and lowest in the phosphate mining area (0.58 ± 0.31 µmol/L). The δ18OP measured in the Huangbai River waters ranged from 6.0 to 20.9‰, with the highest average value in the outcrop area of phosphate rock (average: 14.6‰ ± 3.1‰). The majority of the measured δ18OP values in the Huangbai River deviated greatly from the expected equilibrium values, indicating that δ18OP in this area could be used to trace PO4-P sources. We used two end-member mixing models to quantify the contribution of PO4-P from different sources. In the phosphate mining area, the average fractions of PO4-P from phosphate ore and sewage were 49.5% ± 23.8% and 50.5% ± 23.8%, respectively. In the outcrop area of phosphate rock, the average fractions of PO4-P from phosphate ore and sewage were 60.1% ± 21.7% and 39.9% ± 21.7%, respectively. In the residential/agricultural area, the average fractions of PO4-P from fertilizer and sewage were 49.2% ± 23.2% and 50.8% ± 23.2%, respectively. These results indicate that phosphate mining activities was not an important source for PO4-P in the waters of Huangbai River. The natural weathering of phosphate rock, fertilization, and domestic sewage contributed more to the high PO4-P concentrations in the Huangbai River waters.