Abstract:[Objectives] The Alligator sinensis, classified as a national first-class key protected wild animal, has consistently been a concern for conservationists regarding the improvement of artificial breeding and management standards. This study aims to explore the optimization of feeding patterns for A. sinensis. [Methods] The experiment utilized four different types of feeding:Feeding Group 1 (traditional frozen animal-derived feed), Feeding Group 2 (mixed pellet feed), Feeding Group 3 (mixed pellet feed with chelated sodium butyrate) and Feeding Group 4 (mixed pellet feed with complex Bacillus spores). By collecting feces from each feeding group and analyzing the changes in gut microbiota using 16S rDNA amplicon sequencing, combined with comparative analysis of data such as average clutch size, fertilization rate, and hatching rate of fertilized eggs during the breeding season, the study comprehensively evaluated the impact of different types of feeding on the diversity of gut microbial communities and reproductive performance in A. sinensis. Statistical analysis and data processing were performed using Excel 2010 and SPSS version 26.0. [Results] In terms of intestinal microbial richness and diversity, the intestinal flora of Feeding Group 1 exhibited the highest richness, while Feeding Group 4 had the highest diversity (Table 1). No significant differences were observed in the dominant flora at the phylum level (Firmicutes, Proteobacteria, Bacteroidota, Actinobacteriota) among the different feeding groups of experimental crocodiles. At the genus level, the microbial abundance of Brevibacillus, Cetobacterium, and Paenibacillus in Feeding Group 2, Cetobacterium, Kurthia, and Sphingobacterium in Feeding Group 3, and Brevibacillus, Paenibacillus, and Glutamicibacter in Feeding Group 4 were significantly higher than those in Feeding Group 1 (Fig. 3). The PICRUSt2 functional enrichment analysis revealed that the intestinal flora of Feeding Groups 2 and 4 exhibited the highest relative abundance across fourteen functions, including cell motility, signal transduction, transcription, and amino acid metabolism (Fig. 5). A comparison of the annual egg-laying performance of the four feeding groups of A. sinensis revealed that the numbers of egg-laying nests in Feeding Group 2 (5 nests), Feeding Group 3 (1 nest), and Feeding Group 4 (1 nest) were higher than that in Feeding Group 1 (0 nest). The number of eggs per nest and fertilization rate for Feeding Group 3 were 31 eggs and 91.2%, for Feeding Group 4 were 29 eggs and 89.5%, and the mean number of eggs per nest for Feeding Group 2 were 28.6 ± 4.5 eggs and 85.6% ± 2.3%. After excluding unfertilized eggs, the hatching rate for Feeding Group 4 was 95.9%, for Feeding Group 3 was 92.5%, and the average hatching rate for Feeding Group 2 was 95.3% ± 2.8% (Table 2). [Conclusion] In summary, the mixed particle feed has a positive impact on the intestinal flora structure of A. sinensis, optimizing the composition of intestinal microbes and enhancing the reproductive performance of A. sinensis. This provides a novel approach to the artificial breeding management of A. sinensis, which has the potential to enhance breeding outcomes and the quality of life of A. sinensis.

Abstract:[Objectives] The Chinese Pangolin Manis pentadactyla, currently an endangered species living in the southern region of the Yangtze River in China, may experience various stress factors after being introduced into artificial captive environments, making it susceptible to immune- or metabolism-related diseases. The case analysis of abnormally deceased pangolins in this study aims to provide reference data for the prevention and treatment of similar diseases, thereby improving the health status of captive pangolins. [Methods] This study focuses on the case of a deceased Chinese Pangolin, analyzing the cause of death through autopsy, microbiological tests on tissues and organs, and histopathological analysis using hematoxylin-eosin (H.E) staining. [Results] The body of pangolin was dissected to observe pathological changes. The lungs exhibited obvious congestion with white spots, while no significant abnormalities were found in other organs. Using the VITEK2 bacterial identification system, Citrobacter freundii was detected in the liver tissue, and Serratia fonticola was found in the lungs (Table 1). Pathological analysis revealed severe necrosis and structural disarray in multiple organs (such as the liver, kidneys, lungs, and intestines), indicating rapid disease progression and severe deterioration (Fig. 1). The death of the Chinese Pangolin was caused by infection with C. freundii, which led to an inflammatory response in the digestive organs, affecting digestive function. There was widespread necrosis of columnar epithelial cells in the intestinal mucosa, resulting in symptoms such as loss of appetite and diarrhea. The prolonged symptoms weakened the pangolins immune system, leading to a respiratory infection by S. fonticola in the lungs, where inflammation was pronounced, and the alveolar cavities were filled with proteinaceous edema fluid. The results of the antibiotic susceptibility test showed that C. freundii is more sensitive to fluoroquinolones and aminoglycosides, while exhibiting resistance to penicillin, tetracyclines, macrolides, and cefazolin. S. fonticola was found to be more sensitive to aminoglycosides and moderately sensitive to fluoroquinolones, but resistant to penicillin, tetracyclines, macrolides, and cefazolin (Table 2). [Discussion] Both C. freundii and S. fonticola are opportunistic pathogens. After entering captivity, Chinese Pangolins are often in a stressed state, leading to weakened immunity and increased susceptibility to these opportunistic infections. The unique physiological structure of pangolins, especially their digestive and respiratory systems, makes them highly sensitive to environmental changes, thus more vulnerable to infections. Improving captive conditions to reduce stress responses, closely monitoring for diseases with early interventions, and selecting and using appropriate antibiotics could effectively reduce the risk of infection and improve the survival rate of pangolins. This study will also provide stronger support for future pangolin conservation efforts.

Abstract:[Objectives] Mammals play a crucial role as foundational elements of ecosystems, with their diversity and structure forming the basis of ecosystem functionality. In this study, we analyzed the current status of species diversity, distribution characteristics, and structure of forest-dwelling mammals in the Tangjiahe area of the Giant Panda National Park, Sichuan. We compiled data from 215 infrared-triggered cameras deployed in forest habitats to conduct a preliminary analysis of the altitudinal and seasonal variations in the activity of forest-dwelling species. In addition, we examined the altitudinal distribution and daily activity patterns of the five ungulate species with the highest relative abundance index.Our primary goals included updating species diversity and investigating spatiotemporal distribution patterns which had not been previously reported. [Methods] Data were collected from 215 camera traps deployed in forest habitats ranging from 1 210 to 2 694 m above sea level in the Tangjiahe area of the Giant Panda National Park (Fig. 1). The relative abundance index (RAI, IRA), Shannon-Wiener index (H′), and Pielou’s evenness index (J) were calculated for different altitudinal ranges and seasons. The Kruskal-Wallis test, ANOVA, or t-test were selected based on the normality test for significance analysis. Spearman’s correlation coefficient was calculated between the relative abundance index of five dominant ungulate species and altitude. Daily activity patterns were analyzed using kernel density estimation. All analyses and chart creation were completed using Excel, R 4.2.1, ArcGIS 10.8, and Origin 2022. [Results] The total monitoring effort amounted to 47 196 camera-days, resulting in 73 635 independent photographs. We identified 22 mammal species belonging to 4 orders and 12 families, comprising 3 Primate species, 4 Rodentia species, 8 Carnivora species and 7 Artiodactyla species (Table 1). Among these mammals, 5 species were classified as Class I and 8 species as Class II key protected wildlife in China. Reeves’ Muntjac Muntiacus reevesi, Wild Boar Sus scrofa, Takin Budorcas taxicolor, Chinese Goral Naemorhedus griseus, Tibetan Macaque Macaca thibetana were the 5 most abundant mammal species. Our findings revealed the following characteristics regarding forest mammal diversity in Tangjiahe:(1) The relative abundance index (RAI, IRA) in the high-elevation band (2 101﹣2 700 m, IRA = 570.5 ± 199.5) was significantly lower than that of low-elevation band (1 200﹣1 600 m, IRA = 1 223.8 ± 1 017.8) and middle-elevation band (1 601﹣2 100 m，IRA = 1 355.1 ± 785.1) (Fig. 2). The Shannon-Wiener index and the Pielou’s index were highest at middle elevation band (J = 0.74, H′ = 2.17), and decreased sequentially in low-elevation band (J = 0.60, H′ = 1.82) and high-elevation band (J = 0.59，H′ = 1.77) (Table 2). (2) The species richness in winter (IRA = 936.1 ± 809.6) and spring (IRA = 870.5 ± 598.6) was significantly lower than that in autumn (IRA = 1 366.7 ± 1 087.9, P < 0.05). There were no significant differences in species richness between summer (IRA = 1 178.3 ± 1 032.2) and autumn, spring, or winter (Fig. 3). (3) The Pielou’s index is the highest in summer (J = 0.69), followed by autumn (J = 0.67) and spring (J = 0.67), and is the lowest in winter (J = 0.66). The Shannon-Wiener index was highest in autumn (H′ = 2.05), followed by summer (H′ = 2.03), with the lowest values in spring (H′ = 2.00) and winter (H′ = 2.00) (Table 3). The correlations of relative abundance with altitude for five ungulate species were as follows:Reeves’ Muntjac (ρ =﹣0.6), Wild Boar (ρ =﹣0.25), and Chinese Goral (ρ =﹣0.35) showed a negative correlation with altitude, while Takin (ρ = 0.48) and Tufted Deer Elaphodus cephalophus (ρ = 0.31) showed positive correlations. The relative abundance of the Reeves’ Muntjac was highest at an elevation range of 1 400﹣1 600 m (IRA = 1 318.6 ± 795.4). Wild Boars (IRA = 1 046.5 ± 748.2) and Chinese Goral (IRA = 280.9 ± 117.6) also had the highest relative abundance at 1 400﹣1 600 m. The highest relative abundance for Tufted Deer was found at an elevation range of 1 800﹣2 000 m (IRA = 131.6 ± 85.7). Takin had the highest relative abundance at an elevation range of 2 600﹣2 800 m (IRA = 384.6 ± 123.3). (4) Except for Wild Boar, all species exhibited a bimodal activity pattern, with peaks occurring during dawn and dusk. The peak activity times were 7:00﹣9:00 and 17:00﹣19:00, with a trough from 12:00﹣14:00 (Fig. 5). Significant differences (P < 0.05) were found in the diurnal activity rhythms between the Tufted Deer and the Reeves’ Muntjac, as well as between the Tufted Deer and the Takin. The Wild Boar exhibited a unimodal activity pattern with peak activity from 17:00﹣19:00. [Conclusion] This study shows that making full use of mammal diversity monitoring data can effectively identify species of concern in regional biodiversity inventories, contributing to more objective biodiversity assessments and enabling management agencies to better target conservation and management issues.

Abstract:[Objectives] With the rapid development of the wind power industry, the number of wind turbines has increased rapidly across the country, and the impacts of wind farms on birds have attracted widespread attention. However, there is still a lack of research on assessing the collision risk of birds on wind farms. [Methods] We took Guangxi Xing’an wind farm as an example and investigated avian community and bird collisions within 5 km of the wind farm as well as its surroundings in April-May and September-October 2023 by using the line transect method and the carcass search method. We constructed a collision risk assessment model for birds in wind farms and analyzed its validity by combining bird mortality data in Xing’an wind farm in the past three years and literature review. [Results] The results showed that among the 85 bird species investigated, fourteen species (16.5%) were ranked as extremely high collision-risk, twenty-three (27.1%) ranked as high collision-risk, twenty-two (25.9%) ranked as medium collision-risk, and twenty-six (30.6%) ranked as low collision-risk (Appendix 1). A total of 29 bird strikes were recorded through a combination of field surveys and literature review, including 18 field investigation records and 11 literature collection records, and the birds that had bird strikes belonging to 13 species in 5 orders, 7 families. Among these species, 6 were ranked as extremely high risk, with 19 bird strike fatalities observed, accounting for 65.5% of the total bird strike fatalities; 5 species were ranked as high risk, with 8 bird strike fatalities observed, accounting for 27.6% of the total bird strike fatalities; and 2 species were ranked as medium to low risk, with a total of 2 fatal bird strikes observed, accounting for 6.9% of the total bird strike fatalities (Table 1). [Conclusion] The results indicated that birds of extremely high risk and high risk classes have a higher probability of collision, and the results of the bird collision risk model are consistent with field survey results. This model provides a practical value to assess the risk of bird strikes in wind farms. Its utilization can support bird strike risk assessment for prevention in advance.

Abstract:[Objectives] Bats are crucial for the maintenance of ecosystems, but they also rely on good environmental water sources for their survival. However, there is a lack of research on whether anthropogenically disturbed water environments, particularly urban aquatic ecology and environmental factors, influence bat water utilization. This study aims to provide basic data for the conservation of urban bats by investigating the correlation between aquatic ecology and environmental factors and bat activity in Changchun, China. [Methods] From June to August 2023, bat echolocation calls were recorded at 25 lakes and 6 rivers in the urban area of Changchun (Fig. 1). The study simultaneously recorded 17 ecological and environmental factors of aquatic ecosystems, including those common to both lakes and rivers, such as total number of insects around the water, number of Lepidoptera, Coleoptera, and Diptera in the vicinity, average atmospheric temperature, humidity, and wind speed around the water, chemical oxygen demand, ammonia nitrogen content, pH, dissolved oxygen content, average noise level around the water, and average luminance of the sampling sites. Additionally, lake-specific factors were recorded, including lake area, shape, vegetation coverage around the lake, and the functional zone of the lake. Experiments were conducted for 10 d each in June and July, and for 11 d in August, with each experiment starting at sunset and lasting 3 h. The experiments included acoustic sampling and monitoring of aquatic ecological and environmental factors. One sampling site was selected for each water body, and only one experiment was conducted at each site. In Avisoft-SASlab Lite software, the dominant frequency of bat echolocation calls was used to distinguish bat species. The number of echolocation pulse sequences of bats was used as a proxy for bat activity, and generalized linear models (GLM) and generalized additive models (GAM) were employed to examine the effects of aquatic ecological and environmental factors on bat activity. [Results] In Changchun, four bat species were found in the aquatic environment:Pipistrellus abramus, Hypsugo alaschanicus, Vespertilio sinensis, and Myotis ikonnikovi. M. ikonnikovi was excluded from subsequent analysis due to the relatively low number of echolocation pulse sequences. The average atmospheric temperature around the water was significantly positively correlated with the total number of echolocation pulse sequences for the three bat species (P < 0.001) and H. alaschanicus (P < 0.001). The average atmospheric humidity around the water was significantly positively correlated with the total number of echolocation pulse sequences of the three bat species (P < 0.05) and V. sinensis (P < 0.001). The pH value of the water was significantly negatively correlated with the total number of echolocation pulse sequences of the three bat species (P < 0.001) and H. alaschanicus (P < 0.05). The ammonia nitrogen content in the water was significantly negatively correlated with the echolocation pulse sequences of V. sinensis (P < 0.05). The average luminance of the sampling sites was significantly positively correlated with the echolocation pulse sequences of V. sinensis (P < 0.05, Fig. 2). Lake area was significantly negatively correlated with the total number of echolocation pulse sequences of the three bat species (P < 0.01), H. alaschanicus (P < 0.05), and V. sinensis (P < 0.01). The shape of the Lake was significantly negatively correlated with the echolocation pulse sequences of V. sinensis (P < 0.001, Fig. 3). In lakes in leisure areas, there were significant differences in the total number of echolocation pulse sequences of the three bat species (P < 0.05), H. alaschanicus (P < 0.01), and P. abramus (P < 0.05) compared to residential areas. There was also a significant difference in the echolocation pulse sequences of P. abramus (P < 0.05) in lakes in educational areas compared to residential areas (Table 1). The average total number of echolocation pulse sequences of the three bat species in lakes in residential areas was 2.6 times that in lakes in leisure areas. The average number of echolocation pulse sequences of H. alaschanicus in lakes in residential area was 3.1 times that in lakes in leisure areas. The average number of echolocation pulse sequences of P. abramus in residential area lakes was 3.2 times that in leisure areas lakes and 4.6 times that in educational area lakes (Table 2). [Conclusion] The study results indicate that urban aquatic ecology and environmental factors can influence bat activity. It is recommended to focus on urban water environments to better protect bat diversity in cities.

Abstract:[Objectives] The National Nature Reserve of Guangdong Nanpeng Islands is the only national marine ecological reserve in the eastern Guangdong region, characterized by a rich diversity of marine species, including crabs (Decapoda:Brachyura). However, the status of these crab populations remains largely unknown, hindering effective scientific research and conservation efforts for these species. Therefore, this study aims to conduct a field survey encompassing both the coral reef ecosystem and the shallow continental shelf ecosystem (herein referred to as non-coral reef habitats) to investigate species diversity, community structure, and to update the regional species list of crabs within the National Nature Reserve of Guangdong Nanpeng Islands. [Methods] Field surveys were conducted using trawl and diving methods in June 2023 and June 2024 to assess crab species across 25 sites within the National Nature Reserve of Guangdong Nanpeng Islands (Fig. 1). Of these sites, 10 were located in non-reef areas, while 15 were situated in coral reef regions, representing the majority of crab habitats in this region. We analyzed the structure of the crab community using Pianka’s Index of Relative Importance (IRI), the Hill diversity index, and the Jaccard similarity index. To estimate potential species diversity, we employed statistical extrapolation and interpolation methods. To identify the key species contributing to variations in community composition, we performed SIMPER analysis. Additionally, we conducted non-metric multidimensional saling (NMDS) analysis to explore the spatial distribution patterns of crabs, and utilized one-way analysis of variance (ANOVA) to assess the significance of differences in community structure. [Results] A total of 36 crab species belonging to 12 families and 22 genera were documented in this survey. Species richness was greater in the coral reef area, which harbored 27 species, compared to the non-reef area, which contained 23 species. At the family level, the highest abundance was observed in the Portunidae and Calappidae families, while at the genus level, the Portunus and Calappa genera exhibited the greatest abundance (Fig. 2a). The Portunidae and Xanthidae families demonstrated the highest species diversity, whereas the Charybdis and Portunus genera were the richest at the genus level (Fig. 2b). To evaluate the potential species diversity of the two regions, we assessed sample coverage and conducted statistical extrapolation. The coral reef area exhibited a sample coverage of 87.50%, whereas the non-reef area achieved a coverage of 99.99% (Fig. 3a). Statistical extrapolation indicated that the coral reef could potentially support up to 83 species, which is significantly higher than the estimated 26 species for the non-reef area (Fig. 3b). The non-metric multidimensional scaling (NMDS) analysis revealed significant differences in crab community structure between the coral reef and non-reef areas (ANOSIM:R = 0.540 8, P = 0.001; Jaccard similarity index:0.361) (Fig. 4). The low similarity between the two habitats may be attributed to differences in species composition and the presence of numerous habitat-specific species. For example, the dominant species in the coral reef area was Charybdis acuta, while Portunus sanguinolentus was the dominant species in the non-reef area. In addition, species from the family Euryplacidae were exclusively found in the non-reef area, whereas species from the families Inachidae, Tetraliidae, and Percnidae were exclusively distributed in the coral reef area. By integrating historical records with our survey data, the regional crab species list has been updated to encompass 99 species from 26 families and 66 genera, including 15 newly recorded species. [Conclusion] This study highlights the significance of conducting comprehensive surveys of crab fauna across various habitats, particularly coral reefs. It also provides critical data for the scientific management and conservation of crab resources within the National Nature Reserve of the Guangdong Nanpeng Islands.

Abstract:[Objectives] Two species of foliar nematodes were extracted from the leaves of Tillandsia fasciculata and the root rhizosphere of Vanda subconcolor, both of which were imported from Thailand, using a modified Baermann funnel technique. The taxonomic status of the two intercepted Aphelenchoides populations was determined through comprehensive morphological and molecular analyses. [Methods] For morphological analyses, nematode specimens were extracted, fixed, dehydrated and permanently mounted in anhydrous glycerin on glass slides. Specimens were photographed, measured and illustrated using a Leica DMi8 microscope. Morphometric measurements were carried out and analyzed in accordance with the de Man formula. For molecular analyses, genomic DNA was extracted from individual fresh nematodes. Two sets of universal primer pairs were employed for PCR amplification targeting both the 18S ribosomal DNA (rDNA) and the D2-D3 expansion segment of 28S rDNA. The resulting PCR products were subjected to separation, purification and sequencing. The newly obtained rDNA sequences were edited using DANMAN software package, compared against the NCBI database via BLAST, and subsequently deposited in the NCBI GenBank database. Phylogenetic analyses were conducted using a suite of bioinformatics tools:PhyloSuite v1.2.3 for extracting related species sequences and identifying the best-fit evolutionary model, MAFFT v7.313 for sequence alignment, MrBayes 3.2.7 for constructing Bayesian inference (BI) phylogenetic trees, and iTOL v6 for tree visualization. [Results] The population densities recovered from T. fasciculata and V. subconcolor were approximately 10 ind per 100 g leaves or roots, respectively. Both populations exhibited morphological similarities to A. besseyi, an economically significant plant-parasitic nematodecharacterized by a star-shaped tail tip and classified within the Aphelenchoides Group 3 as defined by Shahina (1996). Comparative morphological and morphometric analyses revealed that the nematode population from T. fasciculata closely resembled the Fragaria × ananassa-associated populationof A. pseudogoodeyi from Florida, USA, exhibiting overlapping ranges for all morphometric data except the b ratio. Similarly, the nematode population from V. subconcolor demonstrated morphological characteristics consistent with the Medicago sativa-associated population of A. medicagus from the United States, with overlapping measurements for all parameters except the c ratios in females. Phylogenetic analyses based on 18S rDNA and the 28S D2-D3 rDNA sequences further demonstrated the distinct evolutionary relationships among the two Aphelenchoides populations. In the 18S BI tree, the population from T. fasciculata formed a well-supported independent subclade (posterior probability, PP = 1.00) with A. pseudogoodeyi, while in the 28S BI tree, it formed a moderately supported independent subclade (PP = 0.98). Concurrently, the population associated with V. subconcolor clustered robustly with A. medicagusin in both the 18S and 28S BI trees, resulting in well supported independent subclades (PP = 1.00). [Conclusion] Based on an integrated analysis of morphological and molecular evidence, the nematode population derived from T. fasciculata was identified as A. pseudogoodeyi, whereas the population from V.subconcolor was identified as A. medicagus. Both of these nematode species represent new geographical records for Thailand, while T. fasciculata and V.subconcolor also serve as new host record species for the two nematodes, respectively. This study significantly enhances our understanding of the ecological characteristics, geographical distribution, and host range of Aphelenchoides nematodes. Furthermore, it offers a scientific foundation for formulating effective plant protection strategies and phytosanitary measures, aimed at mitigating risks associated with importation of plants from Thailand.

Abstract:[Objectives] To investigate the genetic diversity and differentiation among populations of Gymnocypris chilianensis, Schizopygopsis pylzovi,and S. kialingensis distributedin Gansu Province, microsatellite markers were screened and used to assess the conservation genetics of these three species. [Methods]From 2017 to 2022, 309 ind from 16 populations of the three fish species were collected from the Weihe, Taohe, and Daxia Rivers (Yellow River basin); the Bailongjiang River (Jialing River in the Yangtze Valley); and the Shule, Heihe, and Shiyang Rivers (inland river systems). These included 110 ind from 5 populations of G. chilianensis, 72 ind from 6 populations of S. pylzovi, and 127 ind from 5 populations of S. kialingensis. High-throughput sequencing was conducted to identify microsatellite markers in the three species. Polymorphism at microsatellite loci was assessed using samples from the Schizothoracine species, and microsatellite polymorphism was detected by capillary fluorescence electrophoresis. GeneMaker 2.2.0 was used to read the capillary electrophoresis test results. Data were calibrated according to the base repeat unit at each SSR site to obtain genotyping data. Genetic diversity statistics were calculated using PopGene 1.32, including number of alleles (Na), effective number of alleles (Ne), Shannon’s index (I), Nei’s gene diversity (h), and percentage of polymorphic loci (P). Bayesian clustering analysis was performed using Structure software, with the length of the burn-in period parameter and the number of MCMC repeated sampling set to 1 × 10⁵ and 1 × 10⁶, respectively. The K values for the clustering group were set to 1﹣17, with each K value calculated 10 times. A UPGMA dendrogram was generated using Mega 7, with the bootstrap value set at 1 000. [Results] Capillary fluorescence electrophoresis identified 21 highly polymorphic primer pairs suitable for three species (Table 2). These 21 universal primer pairs were used to amplify 309 ind from 16 populations. Among the G. chilianensis populations, Huangyang population showed the highest genetic diversity (Na 1.442, Ne 1.314, I 0.203, h 0.203, P 44.24%), while Zhangye population had the lowest (Na 1.042, Ne 1.042, I 0.029, h 0.021, P 4.24%). For S. pylzovi, Qilian population had the highest genetic diversity (Na 1.539, Ne 1.193, I 0.206, h 0.127, P 53.94%), and Shimen population the lowest (Na 1.236, Ne 1.126, I 0.120, h 0.078, P 23.64%). S. kialingensis showed the highest genetic diversity in the Shimen population (Na 1.351, Ne 1.152, I 0.147, h 0.094, P 35.15%), with the lowest in the Zhouqu population (Na 1.181, Ne 1.089, I 0.088, h 0.057, P 18.18%) (Table 3). Overall, S. pylzovi exhibited higher genetic diversity than G. chilianensis and S. kialingensis. Genetic parameters indicated little intraspecific variation across different populations within species, suggesting comparable genetic diversity levels among populations. Bayesian genotype clustering detected interspecies gene flow between S. pylzovi and G. chilianensis in the Huangyang population, and between S. pylzovi and S. kialingensis in the Shimen population (Fig. 4). A UPGMA dendrogram of 16 Schizothoracine populations based on Nei’s genetic distance showed that S. pylzovi and S. kialingensis populationsformed small branches before clustering into a larger branch. The Shiyang population of S. pylzovi andall G. chilianensis populations formed one distinct branch (Fig. 5). [Conclusion] The genetic diversity of 16 populations across three Schizothoracine species was evaluated, thereby providing valuable insights into their germplasm status and supporting future conservation strategies.

Abstract:[Objectives] The Bider marking is a distinctive feature on horses, characterized by unique and symmetrical black irregular patterns located on the shoulder blades. Currently, this marking is found exclusively on Przewalski’s Horse Equus ferus przewalskii and Mongolian Horses E. caballus that exhibiting a Dun coat color. This study aims to explore the differences in the formation of skin and hair follicles between the Bider marking area (shoulder), the dorsal midline and croup areas of Dun Mongolian Horses. [Methods] Paraffin sectioning and hematoxylin and eosin (H.E) staining techniques were employed to observe and analyze the morphological structure of hair follicles, patterns of pigment deposition, epidermal thickness, and hair follicle density in the skin regions of the shoulder, dorsal midline, and croup of Mongolian Horses. Skin tissue samples were collected from the shoulder, dorsal midlines, and croup of six Dun Mongolian Horses with Bider markings (Fig. 1). These samples underwent paraffin embedding, sectioning (both transverse and longitudinal), H.E staining, and subsequent microscopic observation using a BX53 (LED) semi-electric fluorescence microscope. To assess the symmetry of the anagen hair bulbs, five unique microscopic images of anagen hair bulbs from the shoulder, dorsal midline, and croup were selected through longitudinal sectioning. The hair bulbs were divided into left and right sides based on the central line of the hair papilla. t-test was performed on data exhibiting uniform hair follicle morphology, with data values for the left and right sides of each hair bulb recorded separately. Hair follicle density and epidermal thickness were measured using both transverse and longitudinal sectioning preparation methods. Continuous sections containing hair follicle shafts and the epidermal layer were selected from skin tissues of the shoulder, dorsal midline, and croup, all observed under consistent microscopic conditions. Five representative images were selected for measurements. Hair follicle counting involved tallying the number of clearly visible hair follicles in each selected image, with the total count recorded for each field of view. To measure epidermal thickness, Image View image processing software was employed to obtain precise measurements within the epidermal layer of each image, and the corresponding values were documented. t-test was applied to determine whether significant differences existed in hair follicle density and epidermal thickness across different areas (shoulder, dorsal midline, croup). Statistical analysis of the data was conducted using GraphPad Prism 10 software. [Results] Following longitudinal sectioning and H.E staining of skin tissues (Fig. 2a), the epidermal layer (Fig. 2b), dermal layer, hair follicle structure, and sebaceous glands were examined. The hair follicle structure included the hair shaft, hair bulb (Fig. 2c), hair root, inner root sheath, outer root sheath, and hair papilla. Transverse sectioning facilitated the observation of the hair cortex, medulla, and cuticle structures of the hair shaft (Fig. 2d). Longitudinal sectioning of skin tissues from the shoulder, dorsal midline, and croup (Fig. 3) revealed the structural integrity of the skin tissues and hair follicles. The shoulder (Fig. 3a) and croup (Fig. 3c) were in the anagen phase, while the dorsal hair follicles exhibited atrophy (Fig. 3b), indicating they were in the catagen phase. Pigment deposition was observed in the epidermal layer and hair bulb regions of the dorsal midline, shoulder, and croup. Hematoxylin and eosin (H.E) staining revealed significant melanin deposition in the epidermal and hair bulb areas, facilitating the distinction between eumelanin and pheomelanin. Notably, significant asymmetry in pigment deposition was observed in the anagen hair bulbs of the croup. However, this asymmetry was not observed in the shoulder and dorsal midline (Fig. 4). In the comparative observation of pigment deposition in anagen hair bulb regions (Fig. 5), it was noted that the pigment distribution on the shoulder (P = 0.866) and dorsal midline (P = 0.093) exhibited symmetrical characteristics, with no statistically significant difference in pigment deposition area between the left and right sides. Conversely, pigment deposition on the croup displayed significant asymmetry (P = 0.049). In our comparison of follicle density among the shoulder, dorsal midline, and croup coats (Fig. 6a), we observed highly significant differences in follicle density between the croup and shoulder (P = 0.002) as well as between the dorsal midline and shoulder (P = 0.001). When examining the thickness of the epidermal layers in the shoulder, dorsal midline, and croup coats (Fig. 6b), we found that the difference in epidermal layer thickness between the dorsal midline and croup was not significant (P = 0.558). Additionally, the difference in epidermal layer thickness between the dorsal midline and shoulder was also not significant (P = 0.059). However, a significant difference in epidermal layer thickness was observed between the croup and shoulder (P = 0.020). [Conclusion] This study offered a comprehensive examination of the morphological characteristics of skin and hair follicles, along with pigment deposition patterns, in various coat regions of Dun Mongolian Horses with Bider markings. These findings provide an important foundation and reference point for future research on the morphological basis of the distinctive Bider marking in Mongolian Horses.

Abstract:Two bats were captured (specimen numbers 20180444 and 20190291 respectively) in Guangzhou City and Guangdong Shimentai National Nature Reserve in Guangdong Province in July 2018 and May 2019, respectively. The morphological and skull characteristics of these two bats were measured by using electronic digital calipers and compared with morphological data of Myotis species from other studies. The Cyt b and COI gene sequences of these two bats were sequenced. Then, the Cyt b and COI gene sequences of some Myotis species were downloaded from the NCBI database, and a maximum likelihood phylogenetic tree was constructed using multiple programs in PhyloSuite. Genetic distances based on the Cyt b gene of 25 bats and COI gene of 15 bats were calculated using the Kimura 2-parameter model. Subsequently, phylogenetic trees of multiple species (Myotis and Submyotodon bats) based on Cyt b and COI sequences were then constructed. The main morphological characteristics of the two bats are small body size, with forearm lengths of 35.60 mm and 34.55 mm, respectively. Pelage is soft and thick. Dorsal hairs black basally with buff brown tips. The abdominal hair is slightly lighter than the back hairs. The base hair color is black, and the hair tip is buff (Fig. 1a). The ear is small and narrow, and the tragus is long and straight (Fig. 1b). The wing membrane is black-brown, without any fuzzy covering. The hind-foot length is short, less than half of the tibia length (Table 2). Combined with the skull measurements, they are different from the bats currently named as S. latirostris, S. moupinensis, and S. caliginosus, all of which were previously considered subspecies of M. muricola. For molecular evidence, the Cyt b and COI gene sequences of two bats were amplified by PCR. Based on the results of the phylogenetic analysis, the two captured bats are well clustered with M. muricola from the database. The genetic distances of Cyt b and COI gene are between 1.1%﹣2.1% and 0.9%﹣2.9%, respectively (Appendices 1 and 2). It is found that two bats have a relatively large genetic distance from Submyoodon (Figs. 2 and 3). Based on the phylogenetic analysis of multiple bat species (Fig. 4), the issues between M. muricola and Submyotodon of the classification differences in morphology and phylogenomics, as well as the geographical distribution differences among related species were raised (Fig. 5). In summary, two newly collected bats were identified as M. muricola. We have expanded our understanding of the geographical distribution of this species. The specimens are kept in the Institute of Zoology, Guangdong Academy of Sciences at present.

Abstract:The odd-scaled snake genus Achalinus, widely distributed in northern Vietnam, China, and Japan, is a group of snakes with relatively deficient data. A. dabieshanensis Zhang, Liu, Huang and Zhang, 2023 was first described based on three specimens and is currently known to be found only in Anhui Province, China. During our recent herpetological surveys, an Achalinus specimen (collection number LFR2024132) was collected from Wuning County, Jiangxi Province, China (29°23′21″ N, 114°39′45″ E, 1 470 m a. s. l.). Morphological characteristics of this specimen were examined and measured in calibrated scale (Deli, 0﹣300 mm, accurate to 1 mm) and Stainless steel digital calipers (German Minat, 0﹣150 mm, accurate to 0.01 mm) to primarily identify this specimen, then maximum likelihood tree was constructed using DNAStar, MEGA X, IQ-TREE 1.6.12, and uncorrected p-distance was calculated using MEGA X, based on mitochondrial COI genes, to confirm the species delimitation. The specimen was a subadult female with the following morphological characteristics:(1) dorsal scales strongly keeled, 23 rows throughout the body, the outmost row smooth and significantly enlarged; (2) tail relatively short, the ratio of tail length to total length 0.16; (3) length of suture between internasals shorter than that between prefrontals; (4) supralabials 6, the 4thand 5th in contact with the eye; (5) infralabials 5, first three in contact with anterior chin shields; (6) ventrals 159, subcaudals 44, not paired. Molecular data revealed that the specimen from Wuning County clustered with the type series of A. dabieshanensis, with low intraspecific genetic divergence (0.9%﹣1.3%, Appendix 1). The result of morphological and phylogenetic relationships indicates that A. dabieshanensis represents a new snake record for Jiangxi Province (Figs. 1﹣4, Tables 1﹣3, Appendix 1). This study indicates that A. dabieshanensis represents a new snake record for Jiangxi Province, bringing the total number of Achalinus species recorded in this province to four, and further enriching the reptile diversity of Jiangxi Province. It is recommended that the investigation and research of this species be continuously strengthened, especially in the adjacent areas of known distribution, such as southeastern Hubei Province and northeastern Hunan Province.

Abstract:Collective escape behavior is a complex and rapid response of group-living animals to threats, which is crucial for the survival of animals. In open spaces with limited obstacles, the movement of animal groups is less constrained, and the collective escape can occur in both two-dimensional (e.g., herds) and three-dimensional (e.g., bird flocks and fish schools) spaces. The success of collective escape largely relies on the coordination between individuals in the group. In addition, escaping efficiency can be affected by various factors such as group size and habitat types, depending on the species. In enclosed spaces, movement is restricted, and animals must compete for limited exits and escape routes, often leading to selfish and herd behaviors. Therefore, blocking and clogging may be observed when groups of animals attempt to escape from an exit, resulting in the “faster is slower” effect. Such an effect can be reduced by wider exits and higher levels of coordination. Interestingly, some types of obstacles near the exit may also reduce blocking and increase evacuation efficiency. Even when there are more exits, individuals in the group may follow their predecessors and preferentially use one of the exits over the other ones, leading to the “symmetry breaking” phenomenon. Although collective escape is common and crucial for group-living animals including humans, very few studies have focused on the molecular mechanisms of this behavior. In addition, virtual simulation technology and bionics of collective escape are prospected.

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