The recombinant baculovirus has been widely used as an efficient tool to mediate gene delivery into mammalian cells but has barely been used in fish cells. In the present study, we constructed a recombinant baculovirus containing the dual-promoter cytomegalovirus (CMV) and white spot syndrome virus (WSSV) immediate-early gene 1 (ie1) (WSSV ie1), followed by a puromycin–green fluorescent protein (Puro-GFP, pf) or puromycin–red fluorescent protein (Puro-RFP, pr) cassette, which simultaneously allowed for easy observation, rapid titer determination, drug selection, and exogenous gene expression. This recombinant baculovirus was successfully transduced into fish cells, including Mylopharyngodon piceus bladder (MPB), fin (MPF), and kidney (MPK); Oryzias latipes spermatogonia (SG3); and Danio rerio embryonic fibroblast (ZF4) cells. Stable transgenic cell lines were generated after drug selection, which was further verified by Western blot. A cell monoclonal formation assay proved the stable heredity of transgenic MPB cells. In addition, a recombinant baculovirus containing a pr cassette and four transcription factors for induced pluripotent stem cells (iPSC) was constructed and transduced into ZF4 cells, and these exogenous genes were simultaneously delivered and transcribed efficiently in drug-selected ZF4 cells, proving the practicability of this modified recombinant baculovirus system. We also proved that the WSSV ie1 promoter had robust activity in fish cells in vitro and in vivo. Taken together, this modified recombinant baculovirus can be a favorable transgenic tool to obtain transient or stable transgenic fish cells.

Dam impoundment can cause many adverse impacts on fish assemblages upstream of dams. Here, we investigated fish diversity in one plateau riverine reservoir (Wanfeng Reservoir) using environmental DNA (eDNA) metabarcoding technology. The following conclusions were drawn: (1) 39 species of fish were monitored belonging to 9 orders and 13 families in the Wanfeng Reservoir, most of which were Cypriniformes and included a variety of common farmed fish belonging to Culter, Oreochromis, Acipenser, and Clarias; (2) the fish assemblage structures in the Up (upstream section), Mid (midstream section), and RA (reservoir area section) of this reservoir was significantly different (p-value < 0.01); (3) among the 7 environmental variables that were measured (Tem, TDS, Sal, pH, DO, NH4+, and Tra), DO (p-value < 0.01) and pH (p-value < 0.05) were the main environmental stressors causing differences in fish assemblages in different sites of the Wanfeng Reservoir. This study concluded that dam construction in the karst region provided habitats for the establishment and dispersal of exotic fish.

This work explores the digestive system characteristics of Brachymystax tsinlingensis during early developmental stages and aims to solve the problem of high lethality of fry during the transgression period, which is crucial for the artificial propagation and population conservation of endangered fishes. This study was carried out on intestinal tissue, digestive enzymes, and antioxidant enzyme activities in the early development stage of Brachymystax tsinlingensis. Ten random samples during endogenous nutrition (7, 10, and 11 days after hatching), mixed nutrition (13 and 19 DAH), and exogenous nutrition (31, 33, 39, 45, and 73 DAH) were collected by histological and biochemical analysis methods. The results showed that the intestine of Brachymystax tsinlingensis already has four layers initially at 7 DAH, and the intestinal gland tissue is evident at 73 DAH. The contents of total protein (TP) and the activities of lipase (LPS) and trypsin (TPS) were maximal at 39 DAH, and the activities were 3.20 ± 0.26 mg/mL, 2.52 ± 0.69 U/g, and 2717.45 ± 295.26 U/mg, respectively. Catalase (CAT) and glutathione peroxidase (GSH-PX) activities both showed the lowest values at 39 DAH, which were 0.57 ± 0.11 U/mg and 3.35 ± 0.94 U/mg, respectively. The activity of amylase (AMS) and the content of malonaldehyde (MDA) increased, and the highest values were reached at 45 DAH (1.32 ± 0.41 U/mg) and 73 DAH (1.29 ± 0.43 nmoL/mg), respectively. Superoxide dismutase (SOD) and GSH-PX activities both showed a peak value at 7 DAH (126.58 ± 20.13 U/mg and 6.47 ± 1.86 U/mg). Overall, the changes in intestinal tissue, digestive enzymes, and antioxidant enzyme activities at 39 DAH of Brachymystax tsinlingensis are inseparable from different vegetative stages during the developmental period, and these results can provide a reference for the proliferation and cultivation of Brachymystax tsinlingensis resources.

The Chinese longsnout catfish (Leiocassis longirostris) is an economically important freshwater fish in China; however, its wild resources have declined dramatically in recent decades. Understanding the genetic structure of Chinese longsnout catfish populations is crucial to guide breeding programs and fishy restoration. In this study, 15 highly polymorphic microsatellite DNA loci were used to evaluate its genetic diversity and population structure. Chinese longsnout catfish populations show high genetic diversity; they do not show significant genetic differentiation or systematic geographic pattern of variation. From the upper to the lower reaches of the Yangtze River, the genetic diversity of Chinese longsnout catfish populations showed an increasing trend. The Gezhouba and Three Gorges dams, which physically divide the Yangtze River into upstream and mid-downstream sections, did not contribute to the genetic differentiation of Chinese longsnout catfish populations. Hence, the source of broodstock is not critical for within-river breeding programs and stock enhancement to restore the wild population. In addition, possible effects of dams on differentiation among populations are crucial and long-term evaluation is essential.