Abstract:[Objectives] Microsatellite markers have been widely used in the research of conservation genetics of aquatic animals. However, there is no report on microsatellite loci of Gymnodiptychus pachycheilus. [Methods] From 2014 to 2022, a total of 2 G. pachycheilus populations (56 individuals) were collected from the main stream of the middle Yalong River and Weihe River. Illumina HiSeq sequencing was used to explore microsatellite markers from the genome of G. pachycheilus. Microsatellite loci were tested using 2 wild population samples’ DNA of G. pachycheilus. Null alleles were tested using the software MICRO-CHECKER version 2.2.3, genetic diversity statistics were obtained using AUTOTET. Deviations from Hardy-Weinberg equilibrium and linkage disequilibrium at each locus were calculated using GENEPOP 4.7.5, and significance values were adjusted after applying Bonferroni correction. Polymorphism information content (PIC) value was based on PIC_Calc0.6. [Results] 567 200 microsatellite loci were identified from 85 905 individual sequence reads produced, and 15 primer pairs were designed and testified in the Yalong River Xinlong population and the Weihe River population. In the Xinlong population, the average number of observed alleles per locus was 6.87 (from 3 to 13), the average observed heterozygosities and PIC were 0.712 4 and 0.630 3 respectively, 1 pair linkage disequilibrium was detected and eight loci showed significant deviation from Hardy-Weinberg equilibrium among the loci (Table 2). In the Weihe population, the average number of observed alleles per locus was 8.4 (from 4 to 16), the average observed heterozygosities and PIC were 0.719 5 and 0.680 7 respectively, 4 pairs linkage disequilibrium was detected and 7 loci showed significant deviation from Hardy-Weinberg equilibrium among the loci (Table 3). [Conclusion] These 15 high polymorphism microsatellite loci described for G. pachycheilus will be useful for future study on phylogeography and genetic diversity studies.

Abstract:[Objectives] Human activities have significantly transformed the natural ecological environment of the region, leading to notable impacts on the spatial utilization patterns of local animal populations. Consequently, gaining insights into the spatial requirements of endangered species under conditions of human disturbance becomes crucial for their effective conservation. Home range and habitat utilization serve as paramount indicators of an animal’s spatial needs, and comprehending the characteristics of home ranges and habitat preferences becomes imperative for bird conservation efforts to yield optimal results. [Methods] In this study, we employed the “noose method” to capture individuals and researched the breeding season home range of Reeves’s Pheasants (Syrmaticus reevesii) using satellite tracking. The fixed kernel (FK) density estimation was utilized to estimate the home range and assess the habitat utilization of each individual. By analyzing the percentage of recorded locations within specific land types, we evaluated the habitat utilization of Reeves’s Pheasants. The theoretical habitat utilization was determined by calculating the percentage of each land type in relation to the total study area. To assess the habitat selection preference of Reeves’s Pheasants, we compared the confidence interval of actual habitat utilization with the numerical values of theoretical habitat utilization. Our analysis focused on three key factors associated with human disturbance:the intensity of human disturbance, distance to farm, and distance to residential areas. We performed a stepwise elimination analysis using generalized linear mixed models (GLMM) to examine the factors influencing habitat utilization and home range size of the Reeves’s Pheasants. [Results] From 2020 to 2021, we have collected home range and habitat utilization for 47 individuals. The results showed that the Reeves’s Pheasants mainly utilize mixed coniferous forests, deciduous broadleaf forests, and shrub forests (Table 1). However, when classified according to disturbance intensity, the Reeves’s Pheasants in low-disturbance areas tend to use mixed coniferous forests only (Table 2). In high disturbance areas, the utilization rate of deciduous broadleaf forests by female individuals shows little change with respect to the distance to residential areas and farms, while the utilization rate of mixed coniferous forests increases as the distance to residential areas and farm increases. In low disturbance areas, as the distance to residential areas increases, the utilization rate of deciduous broadleaf forests decreases, but the utilization rate of mixed coniferous forests increases. Additionally, as the distance to the farm increases, the utilization rate of deciduous broadleaf forests also increases, but the relationship between the utilization rate of mixed coniferous forests and the distance to the farm is not as pronounced (Table 3, Fig. 3). The utilization rate of male individuals in deciduous broadleaf forests or mixed coniferous forests is not influenced by disturbance intensity. The utilization rate of male individuals in deciduous broadleaf forests decreases with increasing distance to residential areas and increases with increasing distance to farms. Neither the distance to residential areas nor the distance to farm affects the utilization rate of male individuals in mixed coniferous forests (Table 3, Fig. 4). The home range of females (18.19 ± 1.59 hm², n = 28) was significantly larger than that of males (12.39 ± 1.52 hm², n = 19), and the home range size of females was negatively correlated with the distance to residential areas in the high disturbance area (Table 4, Fig. 5). [Conclusion] These findings enhance our understanding of the spatial requirements and habitat preferences of the Reeves’s Pheasants, shedding light on the influence of human disturbance on their behavior. Our study offers valuable insights into the behavioral responses of Galliformes to ongoing human activities, taking into account factors such as human disturbance, sex, home range, and habitat utilization. Through a comprehensive examination of these factors, our objective is to establish a solid scientific basis for the conservation of ground-nesting forest birds.

Abstract:[Objectives] Exocrine glands distributed throughout the skin are one of the conspicuous characteristics in amphibians. The secretions of these glands play a vital role in reproduction, communication, and defense. In order to investigate the types, distribution, and ultrastructure of the glands in treefrogs, we used histological techniques to observe the microstructure (including Hematoxylin-Eosin, Alcian Blue-Periodic Acid Schiff and Masson staining methods) and ultrastructure of skin samples from cephalic, dorsal, lateral, mental and ventral regions of Hainan Frilled Treefrogs (Kurixalus hainanus). [Methods] A linear mixed effect model was used to analyze differences in thickness of skin, distribution of glands and size of glands among body regions in sex-specific individuals where body regions were used as a fixed factor and individuals were used as a random factor, and sex differences of those metrics in specific body regions where sex was used as a fixed factor and individuals were used as a random factor. All statistics were performed in SPSS 22.0. [Results] Ordinary mucous glands (Ⅰ MG & Ⅱ MG), serous glands, but not specialized glands were found in most areas (Fig. 1). Microstructure analysis revealed that both male and female frogs had the same type of glands, but there were significant differences in the skin thickness, the distribution of glands, and the size of glands among body regions in sex-specific individuals (Fig. 2, 3). In addition, there was a significant sex difference in these metrics in specific body regions (Fig. 2, 3). The differences in skin thickness might result from an adaption to the locomotive and reproductive patterns of K. hainanus. The differences in distribution and size of the glands might be related to defensive function and sexual dimorphism. Ultrastructural analysis showed that there were different sizes and densities of vesicles in mucous glands (Fig. 4), and the aggregate serous vesicles indicated a staged reaction of substance maturation (Fig. 5). [Conclusion] This study provides fundamental information on skin structure, types of glands, and distribution of glands in anurans. Moreover, this study contributes to further study on the chemical component of volatile secretions from K. hainanus with a morphological basis.

Abstract:[Objectives] In order to clarify taxonomy and molecular phylogenetics of Micromys in China,this study analyzed morphological and molecular data of 9 specimens of M. minutus, 22 specimens of M. erythrotis, and 19 pending specimens of Micromys collected between 2018 and 2021. [Methods] Themorphological characteristics of specimens and skulls were described and measured. Molecular phylogenetic studies include the calculation of genetic distance (the Kimura 2-parameter model) and the construction of phylogenetic trees (maximum likelihood, ML). [Results] The morphological characteristics of pending specimens of Micromys showed differences from M. minutus and M. erythrotis. The back of the body is black-brown (Fig. 1). The base of the hair on the abdomen is gray (Fig. 1). The tip of the hair is gray-white (Fig. 1). The color of the hair on the sides of the body is clearly differentiated (Fig. 1), the dorsal surface of the tail is dark brown (Fig. 1) and the ventral surface of the tail is gray-brown (Fig. 1). Tail length is longer than 120% of head and body length (Table 2). The zygomatic arch is clearly curved inward (Fig. 2). Greatest length of the skull and Condyloincisive length (18.59 ± 0.48 mm and 17.43 ± 0.48 mm) is longer (Table 2). Palatal length (9.35 ± 0.11 mm) is longer (Table 2). Braincase height (7.43 ± 0.06 mm) is higher (Table 2). The genetic distances between pending specimens of Micromys and M. minutus and M. erythrotis are 0.115 and 0.136, which are near the genetic distance (0.126) between M. minutus and M. erythrotis. Based on the complete sequence of the Cyt b of mtDNA and nuclear gene sequences (IRBP1, RAG1, and RAG2), two phylogenetic trees were constructed. Samples were clustered into 3 clades in both phylogenetic trees (M. minutus clade, M. erythrotis clade, and pending specimens of Micromys clade) with high confidence (Fig. 4, 5). [Conclusion] The results of morphological analyses and the molecular phylogenetics supported that the pending specimens of Micromys arean independentspecies, corresponding to M. m. pygmaeus reported in previous studies. Based on the locality, genetic distance, and morphological differentiation, it is suggested that the M. m. pygmaeus should be elevated to a species and named M. pygmaeus comb. nov.. The samples of M. minutus in phylogenetic tree based on the Cyt b of mtDNA were clustered into 6 lineages:Japanese and Korean lineage, European lineage, Novosibirsk Russia lineage, Northeast China and Far East Russia lineage, Anhui China lineage, and Taiwan China lineage.

Abstract:[Objectives] Cultivation will lead to the aggravation of desertification and have a serious impact on animal communities. Functional diversity can precisely reflect the differences in community structure caused by environmental or interference stress. In this study, the desert rodent community in Alxa was taken as the research object to explore the changes in functional composition and functional diversity of the desert rodent community under the disturbance of cultivation, to provide a scientific basis for the maintenance of small mammal diversity in desert areas. The functional strategies of rodents to adapt to the environment are different, and their community structure is also different. Functional traits can reflect the interaction between species and abiotic and biotic factors in the environment and are closely related to niche differentiation, species coexistence, and community construction. Environmental filtering significantly affects community functional diversity. To this end, we propose the hypothesis that cultivation will affect the functional diversity of rodent communities and have seasonal changes. Through the change in functional diversity index, it shows the impact on community ecological space utilization, resource utilization, and niche. [Methods] The effects of cultivation and uncultivated on the functional diversity of desert rodent communities in Alxa were studied in April, July, and October of 2018-2020. The trap-day method was used to study rodent communities in disturbed habitats. Five functional traits were selected and quantified:nutrition, life history, physiology, morphology, and activity rhythm. and this paper uses Excel to input, sort out and calculate the species diversity of the original data. The different level of rodent species diversity between cultivation and uncultivated areas was determined by SPSS 20.0 one-way analysis of variance. The significant difference level was set to (P < 0.05) and plotted with Origin Pro 8 software. The ‘mFD’ package of software R 4.2.0 was used to analyze the correlation between the functional axis and species traits, β diversity, Jaccard similarity coefficient, Kruskal- Wallis test, and functional diversity index calculation, and the ‘ggplot2’ package was used for mapping. [Results] The results showed that:(1) In the two communities of cultivation and uncultivated (Table 2), the richness index, diversity index, and evenness index of the cultivation area were higher than those of the uncultivated area, indicating that the species diversity of the cultivation area was the highest and the overall distribution of the species was more uniform, and the cultivation changed the trait composition of the rodent community (Table 3). (2) The community composition in spring, summer, and autumn was significantly correlated with functional traits such as dormancy (P < 0.05), reproductive cycle (P < 0.05), and feeding habit (P < 0.05). The above functional traits could be considered as the main driving factors for the distribution of desert rodents in the Alxa desert (Table 3, Fig. 2). (3) Functional richness reflects the utilization degree of ecological space of the community, functional evenness reflects the utilization degree of effective resources of the community, and functional dispersion reflects the competition among species of the community. There are some spatial differences and seasonal changes in the functional diversity index of the desert rodent community in Alxa. cultivation area:the richness and evenness of community function in spring and autumn were higher than those in uncultivated areas, and the dispersion of community function in each season was significantly higher than that in uncultivated areas. Uncultivated area:the richness of community function in summer is higher than that in the cultivation area, and the evenness of community function in autumn is higher than that in the reclaimed area (Fig. 4﹣6). (4) The highest value of community functional richness in both cultivation and uncultivated areas appeared in summer, and the differences between the two in different seasons were compared (Fig. 3b). The highest value of community functional evenness in the cultivation area appeared in spring (Fig. 4a), and the highest value of functional dispersion appeared in autumn (Fig. 5c), and the difference between the two seasons was small. The highest value of functional evenness of uncultivated communities appeared in autumn (Fig. 4c), and the highest value of functional dispersion appeared in summer (Fig. 5b), both of which were quite different between seasons. (5) Fig. 4:FRic:Functional richness index, The blue dots and the connected parts represent the species distribution points and ranges in the uncultivated areas. the red dot and the connected part represent the species distribution point and range of the cultivation area; Fig. 5:FEve:Functional evenness index; Blue represents the uncultivated area, red represents the cultivated area; the size of the circle represents the abundance of species in the community. the larger the circle is the more species, and the smaller the circle is the fewer species; Fig. 6:FDiv:Functional divergence index; Blue represents the uncultivated area, red represents the cultivation area; the size of the solid circle represents the abundance of species in the community, and the diamond and triangle represent the center of gravity of the community in the uncultivated area and the cultivation area. [Conclusion] The above results indicate that the functional diversity of the community is different due to the differences in community composition and niche. The functional diversity of rodent communities in the Alxa desert area is associated with land reclamation and seasons, cultivation will also affect the functional diversity of the community from the aspects of ecological space utilization, resource utilization, interspecific competition, and niche.

Abstract:[Objectives] Ferritin exists widely in organisms, which can maintain a relatively stable iron content in cells and also participate in body’s immune response. In recent years, due to bacterial infection, the population of Rana amurensis has shown a downward trend. This study explores the expression pattern of ferritin H genes in bacteria-infected R. amurensis, hoping to provide a reference for the study of the mechanism of R. amurensis resistance to bacterial infection. [Methods] In this study, PCR was used to amplify the coding region of ferritin H(ferH) gene of R. amurensis and analyze its bioinformatics. Quantitative Real-time PCR (RT-qPCR) was used to detect the transcriptional changes of ferH gene in the liver, spleen, kidney, skin, and muscle tissue of R. amurensis after Aeromonas hydrophila (Ah) infection. The transcription level of ferHgene relative to the reference gene was calculated by 2^﹣ΔΔCT using EXCEL 2019. Finally, immunofluorescence detection technology was used to analyze the expression of ferH protein of R. amurensis after Ah infection. Three positive regions were selected for each sample to take photos, and Image J software was used to analyze the relative average optical density. The above results were expressed as Mean ± SD. SPSS 26.0 was used for statistical analysis. One-way ANOVA was used to compare the difference between the treatment group and the control group, and the statistical significance was P < 0.05. GraphPad Prism 8 software was used for mapping. [Results] The results showed that the encoding region of ferHgene was 534 bp, encoding 177 amino acids (Fig. 1). The amino acid sequence analysis of this gene showed that it had the highest homology with R. temporaria, reaching 98.37% (Fig. 2). The results of RT-qPCR showed that ferHgene was widely transcribed in R. amurensis tissues (Fig. 3), and the transcription level of ferHgene was significantly up-regulated in liver, spleen, kidney, skin, and muscle tissues after Ah infection (P < 0.01). ferHgene in liver, skin and muscle tissues reached the peak of transcription 6 h after infection, which was 11.95, 24.31, and 24.72 times higher than that in the control group (P < 0.01). The peak transcription in spleen and kidney tissues was 18.22 and 18.19 times higher than that in the control group at 24 h after infection (P < 0.01) (Fig. 4). In addition, immunofluorescence assay results showed that the protein was expressed in the cytoplasm of the liver and muscle tissue of R. amurensis to varying degrees after infection with Ah. ferH protein expression was highest in liver and muscle tissue after Ah 6 h infection, 11.63 and 4.82 times higher than in control group, respectively (P < 0.01) (Fig. 5). [Conclusion] In conclusion, the ferHgene of R. amurensis is up-regulated in response to bacterial infection, suggesting that the gene is involved in bacterial immune response.

Abstract:[Objectives] Saiga Antelope (Saiga tatarica) is a class I wildlife in China and a world-class endangered species. Its origin is mainly in high cold and low oxygen areas, while the existing population mainly inhabits the desert and semi desert grassland in Central Asia. Neuroglobin is a kind of globulin with the ability of blood oxygen transport and storage in vertebrates, which plays an important physiological role in the process of animal adaptation to hypoxia. [Methods] In order to explore the mechanism of tolerance to hypoxia in Saiga Antelope, the distribution and expression of neuroglobin in the heart, liver, spleen, lung and kidney of Saiga Antelope were studied by Immunohistochemistry and qRT-PCR. [Results] The results of Immunohistochemistry showed that NGB protein was distributed in heart, liver, spleen, lung and kidney of Saiga Antelope, and the positive expression of NGB protein was mainly distributed in the cardiomyocytes, liver cells, lymphocytes, alveolar cells and glomerular endothelial cells in the white pulp area of the spleen (Fig. 1). The results of qRT-PCR showed that the expression of NGB in heart, liver, spleen, lung and kidney of Saiga Antelope was different. The expression of spleen was the highest, and the expression of heart was the second, both of which were significantly higher than that of liver, lung and kidney (P < 0.05). The expression of liver was higher than that of kidney and lung (P < 0.05), the difference between the expression of kidney and lung was not significant (P > 0.05), and the expression of lung was the lowest (Fig. 2). [Conclusion] The above studies showed that NGB was positive in the main organs of Saiga Antelope, and the expression of NGB in different viscera organs was different, which indicated that NGB might be involved in the oxygen utilization process of these visceral organs. The specific mechanism will need to be further explored.

Abstract:[Objective] The present study aims to provide a basis for in-depth study of the biological characteristics of White Horse (Equus caballus) and the establishment of species resource. [Methods] In this study, using the male and female Ujimchin White Horse in Inner Mongolia as experimental materials, the primary lineage of ear tissue fibroblasts was established by tissue block adherent culture method, the cell adherent rate, survival rate and growth curve of ear tissue source were studied, and the karyotype map of Ujimchin White Horse was further generated. Data results are expressed as Mean ± SD, Excel was used for data collation and analysis, and all graphs were drawn using GraphPad Prism 5. [Results] The results showed that female and male ear tissues of Ujimchin White Horse fibroblasts were cultured by tissue adhesion and grew into a typical fibroblast morphology, and showed the growth characteristics of “S”. There was significant difference in cell survival rate before and after cell freezing (Fig. 5), but the cells still show good resistance to freezing. According to the karyotyping analysis, the number of chromosome strips of female and male Ujimchin White Horse hair tissue fibroblasts is 2n = 64, of which 31 pairs of autosomes and 1 pair are sex chromosomes. [Conclusion] In this study, the fibroblast cell lines of female and male Ujimchin White Horse hair tissues were successfully established, and the cell lines with good genetic characteristics were obtained and stably cultured, which laid the foundation for subsequent in-depth research.

Abstract:[Objectives] Hibernation is a very frequent occurrence in animals. During hibernation, the activity and digestive functions of animals are reduced, and the metabolic rate of animals is much lower than the basal metabolic rate, especially in amphibians and reptiles. Research demonstrated that the numbers of glucagon, gastrin, and somatostatin cells in the digestive system of Rana catesbeiana during hibernation were higher than that during the non-hibernation period or not significantly changed. Argyrophilic cells are the general term for the endocrine cells of the digestive tract that regulate the digestion, absorption, and feeding behavior of animals. However, little is known about the changes of argyrophilic cells in the digestive tract of R. catesbeiana pre-hibernation and post-hibernation, so we studied these changes during hibernation. [Methods] As a pre-hibernation experimental group, we acquired 10 R. catesbeiana from the Harbin Hada wholesale aquatic products market in September 2020 and acclimated them for 7 d in the laboratory. As a hibernation phase experimental group, 10 R. catesbeiana were obtained in October 2020 and acclimated to lab settings for 7 d in 6 ℃ incubators until mid-December 2020. April 10, 2021, 7 d under acclimatized laboratory conditions for the post-hibernation experimental group. Each R. catesbeiana was fed a live goldfish weighing around 3.5 g every three days while it was being domesticated. The Grimelius silver staining was applied to the intestinal tract. SPSS 23.0 and Duncan’s multiple range test were applied to make a statistical analysis of the argyrophilic cells in the digestive tract of three periods of R. catesbeiana. An independent sample t-test was applied to compare the distribution density of the argyrophilic cells in the same part of the digestive tract of the three groups, and the data were shown as “mean ± SE”. GraphPad Prism Version 8.0 software was used to plot the data. The argyrophilic cells were photographed using MOTICAM ProS5Lite microimaging system. [Results] The argyrophilic cells in R. catesbeiana digestive tract did not differ morphologically during hibernation, pre-hibernation and post-hibernation, being cone-shaped, fusiformis and ellipse. Cone-shaped and fusiformis argyrophilic cells with exocrine activity in various parts of the R. catesbeiana digestive tract had considerably larger densities in the digestive tract during the three periods than elliptical cells with endocrine function (P < 0.01). In all three phases, the distribution density of argyrophilic cells peaked in the jejunum, while the troughs varied. The distribution densities of argyrophilic cells are lowest in the esophagus during the pre-hibernation and post-hibernation periods, and in the cardia during the hibernation period. Except for the pre-hibernation and hibernation periods, there were differences in the distribution density at the pylorus (P > 0.05), and the distribution densities of the esophagus, stomach, duodenum, jejunum, ileum, and rectum during hibernation were significantly higher than those of the pre-hibernation and post-hibernation periods (P < 0.05). In the pre-hibernation and post-hibernation periods, the argyrophilic cells displayed an inverted “U”-shaped distribution pattern, and in the hibernation phase, a “~”-shaped distribution pattern (Fig. 2). [Conclusion] Overall, the variations in the distribution density of argyrophilic cells in R. catesbeiana during the three time periods were correlated with the physiological states and functions of the major digestive tract components. The distribution density of argyrophilic cells increased dramatically during hibernation compared to pre-hibernation and post-hibernation, which was linked to improved immunological response and an increase in the small intestine’s sensitivity to neurotransmitters. In short, the morphological characteristics of the argyrophilic cells did not change significantly in pre-hibernation and post-hibernation, whilst the number of argyrophilic cells changed to accommodate the regular performance of their digestive activities and endocrine functions.

Abstract:[objectives] Fungi are important components of nest microbiota, which are closely related to the survival, reproduction, and environmental adaptation of birds. However, most of the research on the relationship between birds and microbiota is mainly focused on intestinal microbes, but few studies focused on the micro-ecological environment in the nest. So, we want to investigate the fungal community diversity in the nest of the Green-backed Tit (Parus monticolus). [Methods] From April to June 2020, in Aha Lake National Wetland Park in Guiyang, Guizhou Province, the Green-back Tits were attracted by hanging artificial nesting boxes. All nests were inspected once a week in the early breeding period, and once every 2 days after the emergence of nest materials. Microbial samples were collected from successful breeding nests and the nests without laying eggs. Based on internal transcribed spacer (ITS) sequencing technology, the differences in fungal community composition between successful breeding nests and the nests without laying eggs of Green-backed Tit were investigated. [Results] At the phylum level, the results showed that Ascomycota (98.81%) was the main phyla of successful breeding nests, Ascomycota (85.59%) and Basidiomycota (8.33%) were the main phyla of the nests without laying eggs (Fig. 2a). At the genus level, the dominant genera of successful breeding nests were Phialemoniopsis (83.04%), Aspergillus (4.75%), Arthroderma (4.29%) and Scopulariopsis (1.78%), and the dominant genera of the nests without laying eggs were Phialemoniopsis (36.06%), Aspergillus (14.53%), Penicilliu (6.22%), Trichothecium (5.80%), Debaryomyces (1.67%) and Papiliotrema (1.09%) (Fig. 2b). Alpha diversity analysis showed that the diversity and richness of fungi in the nests without laying eggs were significantly higher than that in successful breeding nests (P < 0.05) (Fig. 3). Beta diversity analysis showed that there were significant differences in fungal communities between successful breeding nests and the nests without laying eggs (Fig. 4). A total of 19 biomarkers with statistical differences were detected by LEfSe (Linear discriminant analysis effect size) analysis. The significant biomarkers of successful breeding nests and the nests without laying eggs were distributed in Ascomycota and Basidiomycota, and there were significant differences between these two biomarker species (Fig. 5). [Conclusion] In general, by comparing the differences in fungal composition in successful breeding nests and the nests without laying eggs, we find that birds may affect the composition structure of microbiota in the nests during breeding, showing more potential pathogens in the nests without laying eggs than in successful breeding nests.

Abstract:[Objectives]Temperature adaptation is very important for the survival of animals. However, there is still a lack of correlational research on the temperature adaptation of ectothermic animals such as fish. Perccottus glenii can survive for several days in frozen environments, and Symphysodon aequifasciatus is a widely-reared tropical ornamental fish. These two species of fish were regarded as the representative of the cold-water fish and tropical fish in this study. [Methods] To investigate their temperature tolerance, we performed linear regression on the percentage of experimental fish that were out of balance at different temperatures and then interpolated the temperature value corresponding to 50% individual imbalance to obtain the critical temperature of the two fish species. We also measured their heart rate across the temperature gradient. Then the cardiomyocytes of P. glenii and S. aequifasciatus were acutely isolated, and whole-cell patch-clamp and temperature control techniques were also used to explore the electrophysiological characteristics of voltage-gated sodium channels in two kinds of fish cardiomyocytes at different temperatures (15 ℃, 20 ℃, 25 ℃, and 30 ℃). PatchMaster was used to record the original data, and Igor Pro 6.37 was used to fit the data and draw curves, the significance was inspected using one-way ANOVA. [Results] The temperature tolerance ranges of P. glenii and S. aequifasciatus were﹣2.0 to 27.4 ℃ and 13.1 to 39.3 ℃, respectively. The heart rate of P. glenii increased steadily from 0 ℃ to 19 ℃, reached the highest at 19 ℃, and then decreased gradually (Fig. 3), which was consistent with the electrophysiological characteristics of the voltage-gated sodium channel in the myocardial cells:voltage-gated sodium channels in the cardiomyocytes of P. glenii had the highest peak current (normalized current:﹣0.93 ± 0.11) and the highest opening probability (G50:﹣29.48 ± 0.33 mV) at the temperature of 20 ℃ (Fig. 4 and Fig. 6). The heart rate of S. aequifasciatus increased steadily from 14 ℃ to 31 ℃, which was consistent with the electrophysiological characteristics that the peak current and open probability of voltage-gated sodium channel in cardiomyocytes increased with the increase of temperature within the experimental range between 15 ℃ and 30 ℃, and voltage-gated sodium channels of S. aequifasciatus had the highest peak current (normalized current:﹣1 ± 0) and the highest opening probability (G50:﹣31.46 ± 0.33 mV) at 30 ℃ (Fig. 5 and Fig. 6). [Conclusion] These results indicate that the temperature response of the heart rate and voltage-gated sodium channel complex current in the cardiomyocytes of these two fish species is closely related to their own temperature tolerance range and the temperature of their native habitat. Therefore, the voltage-gated sodium channel of fish cardiomyocytes may play an important role in the temperature adaptation process of the two species of fish and maybe a key protein in the temperature adaptation of the two species of fish.

Abstract:[Objectives] Cipangopaludina chinensis is widely distributed in Asia. It has strong adaptability and can quickly enter a dormancy state under drought.In this paper, we study the dormancy characteristic of C. chinensis under drought stress to provide a research clue for revealing the mechanism of extensive adaptive tolerance, as well as the genetics and breeding of the C. chinensis. [Methods] The samples of C. chinensis were collected in Yancheng. Select individuals with sound ontogeny and consistent size to temporarily keep in the laboratory for 2 days. 30 C. chinensis were placed in an anhydrous plastic bucket for drought stress treatment, and the room temperature was adjusted to 24 ℃. 30 C. chinensis were placed in plastic buckets with water as control, and kept temporarily for 30 days. After 30 days, the liver and kidney tissues of the control group and the stress group were taken at random for 3 biological replications. High throughput sequencing technology was used to sequence and analyze the transcriptome of the liver and kidney tissues of C. chinensis under drought stress. After cleaning the raw sequencing datathen we identified different expressed genes by P < 0.05 and | log2FoldChange | > 2. Finally, GO (gene ontology) annotation and pathway enrichment analysis of the different expressed geneshave been done. [Results] The results showed that under drought stress, 110 genes were up-regulated and 389 genes were down regulated in the kidney tissue, 84 genes were up-regulated, and 86 genes were down-regulated in the liver tissue (Fig. 3), gene ontology is mainly related to the regulation of cell adhesion, extracellular matrix organization, neuron projection regeneration, carbohydrate metabolic process, dephosphorylation, regulation of epithelial cell proliferation, carboxylic acid, organic acid transmembrane transport, and tissue remodeling (Fig. 5); the KEGG enrichment analysis showed that the differently expressed genes were mainly enriched in PI3K-Akt signal pathway, proteoglycan, lactose, sphingolipid biosynthesis and metabolic pathway 5 that are the main pathways under drought stress (Fig. 6). [Conclusion] Based on the enrichment analysis results of GO and KEGG, heat shock protein genes Hsp70, SRCR, SLC28A3, FASN, APMAP, MSTN, Poc1b, S1P and Na⁺- K⁺- ATPase β Subunit 1 have been identified as important genes related to dormancy of the C. chinensis under drought stress.

Abstract:In May 2014, a Jerdon’s Baza Aviceda jerdoni was encountered and photographed during a bird survey in Ehuangzhang, Yangjiang, Guangdong Province (Fig. 1). By reviewing literatures, we confirm that this is the first record of the species in Guangdong Province. In the following years, the species was successively recorded in other areas in Guangdong Province, such as Maoming, Zhanjiang and Shaoguan (Fig. 2, Table 1). Based on the seasonal distributions of these records, we assume that Jerdon’s Baza is a rare resident bird restricted in western and northern montane areas in Guangdong Province.

Abstract:Lycodon pictus was described from Cao Bang Province, Northern Vietnam by Janssen et al. 2019. In 2020, it was confirmed to have a distribution in Nonggang National Nature Reserve, Longzhou County, Guangxi Zhuang Autonomous Region, China, which is the first recorded from China. In September 2021, two snake specimens (one male and one female) of the genus Lycodon were collected in Libo County, Qiannan Buyi and Miao Autonomous Prefecture, Guizhou Province, China (25°14′3″ N, 107°55′32″ E, altitude 530 m) during a field survey. Based on the morphological comparison and phylogenetic analyses, the collected specimens of the genus Lycodon were identified as L. pictus,which is a new record to Guizhou Province. The specimens were fixed in 10% formalin for one day, subsequently kept permanently in 75% ethanol, and deposited in Anhui Normal University Museum (ANUM). The fresh liver tissue was extracted and immediately preserved in 95% ethanol. 31 sequences of 18 recognized species of genus Lycodon and2 outgroups (Boiga cynodon, Dipsadoboa flavida) were downloaded from GenBank (Accession numbers listed in Fig. 2). Uncorrected pairwise distances (p-distance) among closely related congeners were calculated in MEGA X software. Phylogenetic tree was constructed by maximum likelihood (ML) in RaxML 7.2.6 software. After external morphological (Fig. 1) and morphometric measurements (Table 1) examination, the specimens from Libo County were similar to Lycodon pictus. Molecular phylogenetic analyses revealed that the uncorrected pairwise distances (p-distance) between the two collected specimens and L. pictus were from 0.3% to 0.8% (Table 2). The new distribution site of L. pictus in Libo, Guizhou, China is approximately 350 km away from the type locality in Vietnam. The new discovery in the present study expands the distribution area of L. pictus, and it also enriches the species diversity of the genus Lycodon in Guizhou Province.

Abstract:In August 2022, two specimens of amphibia were collected from the Baima Mountain Nature Reserve in Wulong District, Chongqing (Fig. 1 and 2). Through a comprehensive examination of morphological features, it has been identified as Leptobrachella suiyangensis. A molecular phylogenetic analysis was conducted by comparing the 16S rRNA gene sequences of several Leptobrachella species from GenBank with those obtained from the specimens collected in this study. Oreolalax rhodostigmatus and Leptobrachium tengchongense were selected as outgroups (Table 1). The phylogenetic tree was constructed using Bayesian inference (BI) and maximum likelihood (ML) methods. Additionally, the genetic distance between the Leptobrachella species listed in Table 1 was determined using Kimura’s two-parameter model. The Leptobrachella specimen collected from the Baima Mountain Nature Reserve in Wulong District, Chongqing grouped clustered with L. suiyangensis, with a high support rate of 0.98/99 (BI/ML) (Fig. 3). The genetic distance between L. suiyangensis and the type specimen was found to be 1.2% (Table 3), indicating genetic differentiation between the populations from the two locations. The L. suiyangensis discovered in Wulong, Chongqing exhibited a slightly smaller size (23.0﹣23.4 mm) compared to the type specimen with a minimum body length of 28.7 mm (specimen number:GZNU20180606007), and displayed a brownish-yellow dorsal coloration. Based on the combined morphological comparison and phylogenetic analysis, the collected Leptobrachella specimens were identified as L. suiyangensis, representing a novel amphibian distribution record in Chongqing. The study helps to improve our understanding of the species’ distribution patterns and morphological characteristics, and to develop effective strategies for its long-term conservation and management.

Abstract:The genus Nidirana is an amphibian widely distributed in the Oriental realm. Only one species (N. adenopleura) of Nidirana has been recorded in Anhui Province. In July 2022, nine specimens of Nidirana were collected in the Huangshan area of Anhui Province (Jiulongfeng Nature Reserve and Majinling Nature Reserve). Firstly, their morphological characteristics were similar to those of N. mangveni (Fig. 1, Table 2). In addition, the molecular phylogenetic analysis based on mitochondrial COI gene fragments showed that the newly collected specimens had the closest evolutionary relationships with N. mangveni with high support. Moreover, the genetic distance between the new samples and N. mangveni was the closest, with only 0.04% sequence divergence between the new samples and N. mangveni from the type locality (Table 3). Therefore, combined with morphological comparison and molecular phylogenetic analysis, the new specimens were identified as N. mangveni, which is the new amphibian record of Anhui Province, China. This discovery extends the recorded distribution range of N. mangveni to southern Anhui and provides the first-hand basic information for the biogeography of the genus Nidirana.

Abstract:Food caching is an effective survival strategy that animals have evolved to cope with unpredictable environmental changes. Cognition is a major area in ornithological research, with much work being done on avian cognition, particularly regarding food caching behavior. Although spatial cognition has received considerable attention, there has been a dearth of research on social cognition in current studies. Species that heavily rely on stored food during periods of scarcity, not only require well-developed spatial cognition to memorize the location of their caches but also need strong social cognition ability to handle interactions with potential pilferage. This article provides a comprehensive review of social cognition about avian food caching behavior, which focuses on two aspects:social relations cognition and social status cognition. Our review will highlight the social cognitive abilities of birds.

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