Understanding animal movement is a key question in ecology and biodiversity conservation, which is particularly important for the success of reintroduction projects. The movement of critically endangered Chinese giant salamander (Andrias davidianus) remains poorly understood due to the rareness of wild individuals of this species. We lack movement details about the full annual cycle after reintroduction, especially the abiotic and biotic influences that affect its movement. We utilized pilot reintroduction projects as opportunities to fill in some knowledge gaps on their movement ecology. We released 31 juvenile captive-reared Chinese giant salamanders of two age groups in two rivers in the Qinling Mountains of central China and monitored their daily movements for 16 months using surgically implanted radio transmitters. We examined the impacts of individual traits (body mass, body condition) and environmental conditions (temperature, precipitation, and moon phase) on their daily movement patterns. Data were analyzed using a mixed-effects logistic regression model to understand the drivers of their movement tendency (i.e., whether they move or not) and a linear mixed-effects model was used to understand the drivers of their movement distance. We found that movement tendency of the older salamander cohort was positively affected by the moon phase, increasing near the Full Moon, whereas the younger cohort of animals were not impacted by the moon phase. For daily distance moved, we found temperature had a strong positive effect on both cohorts, whereas precipitation had moderate but opposite effects on the two cohorts Body mass and body condition did not have any significant impacts on either age classes’ movement tendency or distance. This study provides insight into the abiotic factors that impact the temporal and spatial movement ecology of reintroduced giant salamander, which will in turn help with designing best practices for future releases and conservation of this iconic montane aquatic predator.

The Chinese giant salamander (Andrias davidianus), an endangered amphibian species endemic to China, has been previously evaluated with regards to its phyletic evolution, zooecology, and ethology, but molecular mechanisms underlying its skin pigmentation remain unknown. Herein, a skin transcriptome database of different colored salamanders was established using RNA-seq, and a total of 47,911 unigenes were functionally annotated. Among these unigenes, a total of 1252 differentially expressed genes (DEGs) were annotated in the seven public databases, and six DEGs were validated by qPCR between five different skin colors and eight tissues. The results showed that TYR, TYRP1, and ASIP were significantly differentially expressed between different body colors, while TYR, TYRP1, and DCT were highly expressed in skin tissue. The full-length complementary DNA of TYR was cloned and analyzed between normal and yellow phenotypes. Three nucleotide sequence deletion sites were identified in the coding region of TYR, leading to premature termination of transcription and translation in yellow individuals. Our study provides useful data for the further study of the molecular mechanisms of melanin formation, and a valuable reference for the breeding of specific skin colors in other salamanders.

Schizothorax prenanti is an economically important cold-water fish in China. Lipopolysaccharide (LPS) can induce an immune response in S. prenanti; however, little is known about the effects of LPS on oxidative stress (OS) and apoptosis in S. prenanti. In this study, S. prenanti fish were stimulated with LPS at a dose of 10 mg/kg of body weight. After 0 h, 12 h and 24 h, the tissue samples were collected. The OS- and apoptosis-related genes and enzymatic activities in the liver, head kidney (HK), and spleen of S. prenanti were analyzed by a two-way repeated-measures analysis of variance (ANOVA). Hematoxylin and eosin and terminal transferase uridyl nick end labeling staining were also performed. In S. prenanti, LPS administration downregulated the catalase (CAT) and B-cell lymphoma/Leukemia-2 (Bcl-2) expression levels, and upregulated BCL2-associated X (Bax) and cysteine-aspartic-specific protease-3 (caspase-3) expression levels. Meanwhile, superoxide dismutase and CAT enzymatic activities were inhibited and malondialdehyde (MDA) content was increased by LPS treatment. Additionally, LPS treatment induced OS damage and apoptosis in tissue sections. These results indicated that apoptosis in the liver, HK, and spleen of LPS-administered S. prenanti may be mediated by OS via the mitochondrial apoptotic signaling pathway. Our findings are expected to contribute to a better understanding of the responses of different tissues to bacterial challenges. In addition, we can increase the tolerance of fish to the OS through dietary manipulation in the future.

The Taibai stream salamander (Batrachuperus taibaiensis) is a recently described species of the genus Batrachuperus that occurs in the Zhouzhi Heihe River and is endangered in its native range. Here, we have established a method for water environmental DNA (eDNA) analysis of Batrachuperus using a series of optimizations. We have designed a specific set of primers for the genus Batrachuperus to amplify a 160 bp fragment of Cytb. The sequences were obtained from nested PCR on eDNA extracted from water samples, after which DNA barcoding was performed according to sequence analysis to determine the presence of the target species in the water. The method was validated using water from the Zhouzhi Heihe River with known B. taibaiensis populations and found that B. taibaiensis eDNA can move at least 150 m downstream from its point of origin. This study is the first to establish an optimal method for obtaining the eDNA of Batrachuperus from water samples, which provides a theoretical basis for resource investigation and the protection of B. taibaiensis in future research. It is also an example of the eDNA extraction of other species that live in similar waters and are less genetically diverse between species.

Heat shock proteins (HSPs) play a key role in anti-stress and immune processes and are associated with autoimmune diseases. In order to explore the immunological role of HSPs from Schizothorax prenanti (S. prenanti), SpHSP60 was cloned for the first time in this study, and the gene expressions of SpHSP27, SpHSP60, SpHSP70 and SpHSP90 in the hepatopancreas, head kidney, hindgut and spleen were analyzed by quantitative real-time PCR (qPCR) after treatment with lipopolysaccharide (LPS). The open reading frame of the SpHSP60 gene (GenBank accession number ON245159) is 1728 bp. It encodes a protein of 575 amino acids. Its C-terminus is a highly conserved and repeated glycine sequence, which is an important cofactor in ATP binding. Compared with the control group, most of the SpHSPs were significantly upregulated in the tissues examined at 12 or 24 h after LPS challenge. The most abundant expression of SpHSP70 was found in the head kidney at 24 h after LPS injection, followed by SpHSP27 in the spleen at 24 h; both of these SpHSPs displayed strong expression under the LPS stresses, about 20–70 fold more than that of SpHSP60 and SpHSP90. The temporal expression patterns of the four SpHSP genes were different in the four tissues examined. Taken together, the results suggest that SpHSP27, SpHSP60, SpHSP70 and SpHSP90 participate in innate immunity stimulated by LPS, and the response intensity of the SpHSPs was organ-specific, indicating they could provide early warning information against bacterial infection. The findings in our study will contribute to better understanding the biological processes and important roles of SpHSPs involved in defending against pathogenic bacterial challenge.

In recent years, the population of Brachymystax lenok tsinlingensis has experienced a rapid decline due to habitat degradation caused by overfishing and dam construction. In the current study, water samples were collected from four tanks containing B. lenok tsinlingensis individuals with wet weights of 5.8 g, 12.3 g, 34.7 g, and 81.7 g, respectively, and the cycle threshold (Ct) values of B. lenok tsinlingensis in each tank were measured using quantitative PCR. Additionally, water samples were collected from six sites along the Zhouzhi Heihe River (Yu Dongquan, Ban Fangzi, Ba Mugou, two sites along Wang Jia, and Qingshui Rivers), and the Ct values of B. lenok tsinlingensis at these sites were quantified. The results show that the relationship between the Ct values measured by the quantitative test and the fish biomass in the corresponding tanks followed an exponential function, with a regression coefficient of 0.9987. The biomass for the six sites was determined using the established relationship, and the results indicate that the biomass of B. lenok tsinlingensis was highest in Yu Dongquan, followed by Ban Fangzi, Ba Mugou, Wang Jia River 2, and Qingshui, with the lowest biomass observed in Wang Jia River 1. These findings are consistent with the species’ adaptive survival distribution across the six sites. This study presents a standardized approach for using an eDNA-based quantitative PCR assay to monitor B. lenok tsinlingensis. It provides an advanced technical method applicable to the protection and monitoring of other aquatic species in future studies.