Abstract:[Objectives] The Lancang River, particularly its downstream Xishuangbanna section in Yunnan Province, represents one of China’s most biodiverse regions for freshwater fish. During several fish biodiversity surveys conducted by the Kunming Institute of Zoology (KIZ) in the Lancang River Basin in Xishuangbanna Dai Autonomous Prefecture, Yunnan Province, between 2015 and 2025, specimens of four newly recorded indigenous species and an exotic species were collected. [Methods] In this study, key diagnostic characteristics of the collected specimens were measured, and morphological characteristics were compared against relevant taxonomic literature. Specimens were taxonomically identified through comprehensive morphological analysis. [Results] In this article, the five newly recorded species are described, and morphological photos (Figs. 1﹣6) and data (Tables 1﹣4) for identification are provided. Based on the morphological evidence, the specimens of the four newly recorded indigenous species were identified as Pseudecheneis sympelvicus, Gyrinocheilus pennocki, Cosmochilus harmandi,and Labeo chrysophekadion, and the specimens of the newly recorded exotic species were identified as L. catla.[Conclusion] These five species are the first records in China.This study provides basic data for identification of fish species in the lower Lancang River Basin of China and a scientific basis for the investigation and conservation of fish resources in the Xishuangbanna area.

Abstract:[Objectives] Niche differentiation is a fundamental mechanism enabling the coexistence of sympatric species within shared habitats. This study investigates the niche differentiation strategies of four small and medium-sized carnivores—Viverricula indica, Prionodon pardicolor, Paguma larvata, and Prionailurus bengalensis—in the Baixi Provincial Nature Reserve, Zijin County, Guangdong. By analyzing their spatial and temporal niche dynamics, this work aims to inform conservation strategies for these endangered species. [Methods] From July 2020 to August 2022, camera trapping surveys were conducted to monitor four sympatric small and medium-sized carnivores in the Baixi Nature Reserve, with 72 camera traps placed in 51 grids of 1 km × 1 km. Spatial niche differentiation was quantified based on the spatial overlap index (IO). Kernel density estimation (KDE) was employed to model the diel activity patterns of the four carnivores, and overlapping coefficients (Δ) were calculated to compare the diel activity patterns among the four carnivores. Then, Wald tests were conducted to assess whether there were statistically significant differences in diel activity patterns among the four carnivores. The annual activity patterns of the four carnivores were assessed by the relative abundance index (IRA). Then, we performed Pearson correlation tests to assess the similarity in annual activity rhythms among the four carnivores. All statistical analyses were conducted in R 4.2.1. [Results] (1) In terms of spatial distribution, the IO values between V. indica and the other three small carnivores were lower (ranging from 0.20 to 0.38), while those among Prionodon pardicolor, Paguma larvata, and Prionailurus bengalensis were higher (ranging from 0.46 to 0.71) (Table 2). All the four carnivores showed a clear preference for evergreen broad-leaved forests, while Paguma larvata and Prionailurus bengalensis possessed more records of activities in other forests and shrubs (Fig. 3). (2) The diel activity pattern of V. indica significantly overlapped with those of Paguma larvata (Δ = 0.87) and Prionodon pardicolor (Δ = 0.83). Prionailurus bengalensis showed a different diel activity pattern compared with the other three carnivores (Δ ranging from 0.64 to 0.73, P < 0.01, Fig. 4). (3) The annual activity rhythms of the four animal species were fairly similar, while there were differences in the months when their peak activity occurred (Fig. 5). [Conclusion] The findings demonstrated that sympatric carnivores in the study area exhibited significant habitat preference overlap within comparable vegetation communities, resulting in pronounced interspecific competition pressures. This investigation provided novel insights into the temporal activity patterns and niche differentiation mechanisms among small and medium-sized carnivore guilds in South China, offering critical scientific support for developing regionally specific conservation strategies targeting endangered species within increasingly fragmented ecosystems.

Abstract:[Objectives] Activity rhythms and time budgets are not fixed throughout the life cycle of animals. Instead, animals exhibit adaptability across various dimensions. This adaptability, particularly with respect to the temporal aspects of ecological niches, serves as a viable mechanism for animals to respond to cyclical environmental changes. However, the effects of annual and seasonal cycles on animal activity in the tropics remain unclear, and it is uncertain whether animal activity is plastic. Muntiacus nigripes, a diminutive deer species, inhabits tropical forest ecosystems, yet its natural ecology remains largely unexplored. Therefore, this study examined variations in the activity rhythm and time budget of M. nigripes across different temporal scales. [Methods] From November 2020 to October 2022, 160 infrared-triggered camera traps were deployed in the Jianfengling forest area of National Park of Hainan Tropical Rain Forest, Hainan Province, China to monitor M. nigripes and other wildlife. Based on the recorded infrared camera data, the kernel density estimation method built into the “overlap” package was employed to draw the daily activity rhythm curves of M. nigripes in different years, seasons, and sexes. In addition, we used the “overlapEst” function to calculate the overlap coefficient, and applied the Wald test in the “activity” package to assess the statistical significance of difference of the daily activity rhythm in M. nigripes. We recorded the behaviors of M. nigripes through photos and videos. The time budget of daily activity was expressed as the ratio of the number of samples with different behaviors to the total number of samples. The frequency ratio of daily behavioral time allocation was expressed as the ratio of the number of samples of a behavior in a time period to the total number of samples of all behaviors in that time period. Furthermore, we performed the Mann-Whitney U test to examine differences in time budgets for different daily behaviors between different sexes and between different seasons. All analyses were performed by R 4.2.3. [Results] A total of 2 739 independently valid photos were captured by 160 cameras over 45 719 working days. The results showed that M. nigripes was a diurnal species, mainly active at dawn and dusk.Its daily activity rhythm varied between years and seasons, and the overlap coefficient of daily activity was different between 2021 and 2022 (P < 0.05). Among the seasonal changes of different individuals, only the overlap coefficient of daily activity of male individuals in 2021 showed a seasonal difference (P < 0.05), while the rest showed no significant difference (Fig. 3). The temporal dimension of the ecological niche of M. nigripes showed obvious inter-annual variation in social isolation, with 2021 standing out as a significant period of differentiation. Specifically, during the dry season of 2021, a significant divergence was observed in the overlap coefficient of daily activities between males and females (P < 0.05). Similarly, during the rainy season of 2021, a significant difference was observed in the overlap coefficient of daily activities between sexes (P < 0.05). Conversely, no significant difference was noted in the overlap coefficient of daily activities between males and females in 2022 (Fig. 4). The primary behaviors of M. nigripes were locomotion and feeding, which respectively accounted for 36.2% and 48.1% of the total behaviors of females and 44.6% and 39.7% of the total behaviors of males (Fig. 5). In addition, sexes influenced the diurnal time budget for feeding, grooming, and parental care behaviors (P < 0.05), and seasons influenced feeding behaviors (P < 0.05). [Conclusion] In the natural ecological setting devoid of large carnivores, M. nigripes exhibits signals of releasing activity times throughout the day, demonstrating plasticity in activity rhythm and time budget across sexes and seasons.

Abstract:[Objectives] Birds play a crucial role as indicators of the ecological environment. The species composition and its fluctuations serve as significant factors in the conservation of biodiversity and the management of habitats. [Methods] In this study, we conducted a comprehensive analysis of bird records obtained from our investigation, ‘BirdReport’, and relevant literature. To update and analyze the diversity of birds in Shanxi, we referred to A Checklist on the Classification and Distribution of the Birds of China (Fourth edition). [Results] Shanxi Province boasted high bird diversity, harboring a total of 457 species belonging to 76 families of 21 orders. Among these species, 13 species are endemic to China, highlighting their unique presence within the region. In addition, 25 are classified as class Ⅰ national key protected animals, while 80 are categorized as class Ⅱ national key protected animals. Furthermore, 13 and 53 species are listed in Appendixes Ⅰ and II, respectively, of the Convention on International Trade in Endangered Species of Wild Fauna and Flora (CITES). According to the IUCN Red List of Threatened Species, 3, 7, and 17 species are critically endangered (CR), endangered (EN), and vulnerable (VU), respectively (Table 1). The birds in Shanxi exhibited diverse patterns of residence and migration, with a breakdown of 99 resident birds, 151 summer visitors, 77 winter visitors, 115 migrants, and 15 vagrants. Among the 250 breeding species, Palearctic species (103 species) (Appendix 1) are slightly dominant, which indicates a mixing and transitional avifauna realm within Shanxi. [Conclusion] This study presents a systematic analysis of the bird species composition, endangered status, residency types, and realm characteristics in Shanxi Province. The results contribute a scientific foundation to conservation management and further research in the region.

Abstract:[Objectives] The reproduction management of captive female Giant Pandas Ailuropoda melanoleuca is crucial for ex-situ conservation in Giant Pandas. The primary difficulty in reproduction management is to determine when female Giant Pandas are in estrus. However, the current methods for determining estrus have many limitations, such as susceptibility to observers’ experience or other subjective factors, complicated operation, and high costs. This study investigated the changes in daily fecal weight of 30 captive female Giant Pandas during the 31-d pre-mating period (31 d before the day prior to natural mating or artificial insemination) and analyzed their relationship with the estrus initiation, as indicated by changes in estrogen and progesterone levels, aiming to provide a new perspective for assessing estrus initiation and improving reproduction management. [Methods] A total of 1 168fecal samples from 30 captive female pandas (aged 5﹣20 years old) over 38 pre-mating periods were collected daily. The samples obtained at 15:00 and 09:00 the next morning were combined and weighed together as the daily fecal weight by a TCS-150 electronic platform scale (capacity:150 kg, division:10 g, accuracy:± 10 g). Urinary estrogen and progesterone levels were monitored via enzyme-linked immunosorbent assay (ELISA) to determine the date of hormone initiation. Temporal patterns of changes in fecal weight were modeled through 5th-degree polynomial regression (Python 3.10.9 with numpy 1.23.5), with elbow points of the weight decrease identified via the KneeLocator method (Python 3.10.9 with kneed 0.8.3). We employed the Wilcoxon signed-rank test (Python 3.10.9 with scipy 1.10.0) to statistically analyze the changes in fecal weight, with a significance level set at P < 0.05. [Results] The results showed that (1) the elbow point of fecal weight decrease (17.5 ± 4.8 d before mating) appeared about 8 d earlier than the hormone initiation (9.2 ± 2.3 d before mating) (T = 6.5, P = 2.0 × 10^﹣7). Fig. 2 presented the polynomial curve fitting of changes in fecal weight in the 38 pre-mating periods, and Fig. 2 and Table 1 indicated the elbow points and the time points of hormone initiation; (2) the time of the fastest fecal weight decrease (9.8 ± 5.6 d before mating) between the elbow point and the 3rd day prior to mating coincided with the hormone initiation time, with no significant statistical difference observed (T = 313.5, P = 0.99). Table 1 detailed the time points of the fastest fecal weight decrease and hormone initiation; and (3) there was a significant difference in the standard deviation of fecal weights that could be used to distinguish whether Giant Pandas gave birth or not after mating. Specifically, during the phase of stable fecal weight (31﹣18 d before mating) and the phase of fecal weight decrease (17﹣1 d before mating), the P-values were 2.3 × 10^﹣3 and 1.5 × 10^﹣5, respectively. Fig. 3 illustrated the mean values and standard deviations of daily fecal weight during the 31-d pre-mating period for Giant Pandas that would give birth and those that would not, and Table 2 presented the statistical significance in fecal weight between the two groups. [Conclusion] These results provide new perspectives and methods for assessing the timing of estrus initiation and reproductive outcomes in captive female Giant Pandas, which will help to expand the preparation time for reproduction management and improve reproduction success of captive Giant Pandas through precise resource allocation.

Abstract:[Objectives] Food serves as the energy foundation for the survival and reproduction of animals, and its nutritional quality is of utmost significance for the development of animal individuals and populations. This study aims to evaluate the quality of bamboo, the staple food of Giant Pandas, in the Liziping National Nature Reserve, Sichuan Province. [Methods] The Liziping National Nature Reserve serves as both the distribution area and the release site for the small population of Giant Pandas Ailuropoda melanoleuca. The nutrient-to-energy ratios of protein, carbohydrate, and fat in the stems, leaves, and shoots of two bamboo species (Arundinaria spanostachya and Yushania lineolata) in this area were compared using the nutritional geometric model. [Results] There was no significant difference in the nutrient-to-energy ratio of nutrient provided by each vegetative organ between the two bamboo species. The bamboo shoots and leaves had significantly higher protein-to-energy ratios than stems, supplying more than half of the energy for Giant Pandas (Fig. 3, Table 1). Bamboo stems, especially those of multiple years, had a higher carbohydrate-to-energy ratio and could be used as an important energy source for Giant Pandas in winter and spring. The age of bamboo had no significant impact on the nutrient-to-energy ratio in leaves (Figs. 4, 5). The nutrient-to-energy ratios provided by the two bamboo species for Giant Pandas in the study area were similar to those in other distribution areas and could meet the energy demands of both wild and released Giant Panda individuals. [Conclusion] This study evaluates the quality of bamboo in the Liziping National Nature Reserve from the perspective of energy balance. It provides crucial basic information for the protection, restoration, and management of the Giant Panda habitat, particularly the bamboo forest, in this region.

Abstract:[Objectives] Sinospelaeobdella wulingensis and Haemadipsa tianmushana are two species of terrestrial blood-sucking leeches with roughly sympatric distribution in the Wuling Mountain region and surrounding ranges. The former inhabits karstic caves while the latter resides in open environments outside caves, thus facing distinct microhabitat conditions and food availability. [Methods] In this study, 2.5% glutaraldehyde fixing solution and anhydrous ethanol gradient dehydration were employed to prepare the samples of S. wulingensis from Shuiniu Cave in Jishou City, Hunan Province and the samples of H. tianmushana from Hunan Baiyunshan National Nature Reserve in Baojing County, western Hunan Province. The oral suckers of the two leech species were scanned and photographed by scanning electron microscopy (SEM). Image J, Excel, and SPSS 27.0 were used for data measurement and statistical analysis of the observed receptors. The normality of the measured data was assessed by the Shapiro-Wilk test, and the homogeneity of variance was evaluated by the Levene’s test. For comparisons between groups, the t-test was employed. All data were expressed as mean ± SD, and P < 0.05 was considered statistically significant. [Results] The chemoreceptors associated with foraging and feeding of the two leech species were remarkably similar to those found in aquatic leeches. On the dorsal margin of the upper lip of the oral sucker of S. wulingensis, approximately 70 round large chemoreceptors with the diameter of 18.04 ± 2.08 μm were distributed in a band, corresponding to the radial ribs on the oral sucker, and the central regions of these receptors contained either solitary or multiple-in-bundle sensory cilia (Fig. 1). In H. tianmushana, two types of circular chemoreceptors were identified—similar in morphology but different in size, with the central regions of these receptors containing either solitary or multiple-in-bundle sensory cilia (Figs. 2, 3). The large chemoreceptors had the diameter of 17.28 ± 1.53 μm, with similar shape, size, and distribution as those in the dorsal margin of the upper lip of S. wulingensis, and their number was about 90 (Fig. 2). In contrast, small receptors, with the diameter of 8.08 ± 0.48 μm, were observed mainly in the oral sucker and its adjacent epidermis (Fig. 3). In addition, some pore structures (measuring 0.78 ~ 1.15 μm, possibly involved in mucus secretion) were observed on the oral sucker epidermis of both species. [Conclusion] The number of large chemoreceptors in S. wulingensis is lower than that in H. tianmushana, which may be the result of long-term adaptation to the cave environment (Figs. 1﹣3). From an ultrastructural perspective, the chemoreceptors associated with foraging in the oral suckers of these two terrestrial blood-sucking leeches are examined and compared, which provides essential data for future studies on functional morphology, foraging behavioral ecology, and adaptive evolution of leeches.

Abstract:[Objectives] To understand the species of Dactylogyrus that infect Gobio acutipinnatus in the Irtysh River in Xinjiang, China, this study collected 510 ind of G. acutipinnatus from the Haba River (a tributary of the Irtysh River) between July 2021 and August 2023. [Methods] Morphological methods and molecular biological techniques were employed to identify the Dactylogyrus species collected from the gills of G. acutipinnatus. The morphological characteristics of this species were illustrated, measured, and compared with previously reported morphological data of Dactylogyrus parasitizing G. acutipinnatus from both domestic and international sources. The 18S-ITS1-5.8S rDNA sequences were amplified, and sequence alignment was performed by BLAST in the NCBI database. Based on the 18S-ITS1-5.8S rDNA sequences, the base composition was calculated by MEGA 7.0.20, and the Kimura-2-parameter (K2P) model was adopted to calculate the genetic distances between species. PhyloSuite was used for multiple sequence alignment, and the best-fit model was selected through Akaike Information Criterion (AIC) testing with ModelFinder. Based on the selected model, phylogenetic trees were constructed by both Bayesian inference (BI) and maximum likelihood (ML) methods to validate the phylogenetic relationships of parasitic Dactylogyrus. [Results] The collected Dactylogyrus species had a straight line-shaped dorsal bar, with two ends protruding backward and slightly wider in the middle. The ventral bar is straight line-shaped, with narrow ends, extending to the front and slightly wider in the middle. The copulatory tube is a thick bend, with the thickest starting part and a thin tail. The accessory piece is thin tubular, starting from the top of the copulatory tube and bifurcating at the tail end, forming long and short sheet branches at both sides of the copulatory tube (Figs. 1, 2). Consistent with the D. gobii parasitizing G. acutipinnatus of Kazakhstan in the copulatory organ and opisthaptor, the D.gobii parasitizing gills of the G. acutipinnatus inhabiting the Irtysh River, China was identified as D. gobii according to morphological characteristics (Table 3). The characteristic structure of this parasite differed from those of other Dactylogyrus species parasitizing Gobio. The opisthaptor shape of D. gobii is most similar to that of D. cryptomeres, varying only in the ratio of inner root and outer root for anchor, which was 4:3 for D. gobii and 2:1 for D. cryptomeres. The copulatory organs of both species of Dactylogyrus are curved tubular structures. In D. gobii, the accessory piece is in the form of a thin tube, starting from the tip of the copulatory organ and bifurcating into two branches, one long and one short, in the middle and rear part of the copulatory tube. For D. cryptomeres, the copulatory tube was relatively thick, with an enlarged top, and the accessory piece is in the shape of a flake-like mass, with the terminal end forming a sickle shape. D. gobii, D. cristatus, D. finitimus, and D. squameus possessed the D. extensus type of anchor, while only the anchor of D. gobiocypris was of the D. wunderi type. In addition, there are differences in the copulatory organ between D. gobii and D. gobiocypris. The accessory piece of the latter consists of two parts:one is horseshoe-shaped and the other is semi-saccular (Table 3). The18S-ITS1-5.8SrDNA sequence of D. gobii showed the highest similarity of 94.04% to that of D. cryptomeres. The genetic distances based on the 18S-ITS1-5.8S rDNA sequence between D. gobii and nine other species of Dactylogyrus varied within the range of 0.039﹣0.268, and D. gobii had the closest genetic distance (0.039) to the D. cryptomeres from G. gobio (Table 2). The phylogenetic trees indicated that the D. gobii determined in this study clustered into one clade with D. cryptomeres, D. finitimus, D. squamous, and D. gobiocypris parasitizing Gobioninae (Fig. 3). [Conclusion] The results showed that the D.gobii was a new record species of Dactylogyrus in China.

Abstract:[Objectives] This article aims to explore the adaptive mechanism of Tibetan Sheep Ovis arise epididymis in high-altitude environments. [Methods] The epididymal tissue and semen samples were collected from Tibetan Sheep in Haiyan County (at an altitude of approximately 3 500 m) and Xunhua County (at an altitude of approximately 2 300 m) in Qinghai Province. Sperm extraction, paraffin embedding and tissue sectioning, fluorescence quantitative PCR, Western blotting, and immunohistochemistry were employed to compare semen quality, epididymal histology, and expression and distribution of sperm maturation- associated genes PRDX2, STAR, and GALNTL5. The 2^﹣ΔΔCt method was adopted to calculate the relative mRNA level, and Western blot results were analyzed by Image J. The data were analyzed in SPSS 25.0. Independent sample t-test was performed to compare gene and protein expression levels between two altitudes, and one-way analysis of variance and multiple comparisons were performed among three different tissues. The results are expressed as mean ± SD. P < 0.05 indicates a significant difference. [Results] The semen quality of high-altitude Tibetan Sheep slightly decreased, while meeting the fertilization criteria (Table 3). High-altitude Tibetan Sheep had a smaller inner diameter of the lumen and thicker columnar epithelium at the tail of the epididymis (Table 4). PRDX2 was mainly expressed in epididymal epithelial cells, and its expression level was down-regulated in the epididymis of high-altitude Tibetan Sheep. STAR mainly existed in the principal cells of the epididymal epithelium, with up-regulated expression in the epididymis of high-altitude Tibetan Sheep. GALNTL5 was mainly located in the main and basal cells of the epididymis, and its expression level in the epididymis was up-regulated in high-altitude Tibetan Sheep (Figs. 2﹣4). [Conclusion] The differences in histology of epididymal tissue and semen quality between Tibetan Sheep at different altitudes are adaptive changes to high-altitude and low-oxygen environments. The down-regulated expression of PRDX2 as an antioxidant for sperm in the epididymis of high-altitude Tibetan Sheep is hypothesized to be due to the decrease in oxygen content in the epididymal environment under low-oxygen conditions. The upregulated expression of STAR in the epididymis of high-altitude Tibetan Sheep indicates that this protein promotes the synthesis of androgens to maintain the stability of the epididymal lumen environment under high-altitude and low-oxygen conditions. GALNTL5 as a starting enzyme for catalyzing the synthesis of glycoproteins is upregulated in the epididymis of high-altitude Tibetan Sheep, indicating that this protein promotes the synthesis of glycoproteins in the high-altitude and low-oxygen environment to maintain sperm maturation in the epididymis.

Abstract:The Pygmy Cormorant Microcarbo pygmaeus is the most diminutive member of the cormorant family. It had not been sighted in China for many years until it was rediscovered at the Manas Wetland in Xinjiang in 2018, but no breeding records had been observed yet. From May 20 to June 23, 2024, we used a drone to discover 25 Pygmy Cormorant nests, containing 121 chicks in the reed wetlands of Nalati National Wetland Park, Xinjiang (Figs. 1, 2). The average brood size was 3.9 ± 1.1 chicks (n = 15), and the average distance between nests was 30.39 ± 1.35 m (n = 24). Using Nearest Neighbor Analysis, we analyzed the spatial distribution characteristics of the nests. The analysis revealed that the nesting sites of Pygmy Cormorants exhibiting an aggregated distribution pattern within the species, while showing a significant avoidance pattern when compared to Great Cormorant Phalacrocorax carbo, which breeds in the same area (Tables 2, 3, Fig. 3). This discovery marks the first recorded breeding occurrence of the Pygmy Cormorant in China, and highlights the Nalati National Wetland Park as a critical breeding habitat for this species. Further ecological monitoring and conservation efforts are urgently needed to support the breeding population in this region.

Abstract:In August 2023, a female Myotis bat was captured by a harp trap in Shangcheng County, Xinyang City, Henan Province. The echolocation calls of the specimen were recorded during its stationary and exploratory behaviors. The external morphology and cranial parameters were measured, and genomic DNA was extracted from the wing membrane tissue. The complete sequence of the ND1 gene was amplified to facilitate the construction of a phylogenetic tree for species identification. Morphometric analysis indicated that the specimen was relatively large, with a forearm length of 64.3 mm. The dorsal fur exhibited a dark gray-brown color with nearly black roots, while the ventral fur was gray (Fig. 1a). The tragus was narrow and elongated, measuring 11.1 mm, approximately half the length of the ear (Fig. 1b). The wing membrane originated at the base of toes, and the tail membrane commenced at the ankle joint (Fig. 1c). The skull was slightly flattened, featuring a prominent sagittal crest (Fig. 2). The echolocation calls were characterized by a frequency-modulated type, comprising two harmonics (Fig. 3), with a peak frequency of 44.1 ± 4.9 kHz. The maximum likelihood phylogenetic tree constructed based on the ND1 gene (Fig. 4) indicated that the Myotis specimen from Henan shared the same clade with M. chinensis (GenBank accession number:AB106567), exhibiting minimal sequence divergence and a genetic distance of 0.003. Conversely, the genetic distances between the Henan specimen and both M. blythii and M. pilosus exceeded 0.120. According to the data from external morphology, cranial measurements, and the phylogenetic analysis, the Myotis specimen from Henan was identified as M. chinensis, marking a novel record of bat species distribution in Henan Province.

Abstract:A snake specimen was collected from a concrete road in Hejiang County (106°16′33″ E, 28°39′06″ N, 696 m above sea level), Luzhou City, Sichuan Province, China, during a survey on the diversity of amphibians and reptiles in June 2023 (Fig. 1). The morphological characteristics of this specimen were similar to those of Longsheng Kukri Snake Oligodon lungshenensis (Fig. 2). On the phylogenetic tree built based on mitochondrial COI gene sequences, the specimen formed a highly supported monophyletic group with Longsheng Kukri Snake (with a Bayesian posterior probability of 0.99) (Fig. 3), and the genetic distance between the two was 1.6% (Appendix 1). The genetic distances between the newly collected specimen and other congeneric species ranged from 3.4% to 16.8%, and those between Oligodon and the outgroup Lycodon gongshan varied between 17.2% and 20.5% (Appendix 1). According to the results from both morphological comparison and phylogenetic analysis, we confirmed that the specimen was O. lungshenensis, representing a new record in Sichuan Province.

Abstract:Bufo rubroventromaculatus is a newly discovered species of the genus Bufo from northern and central Vietnam in 2024. Currently, its distribution in China has been recorded only in Napo County, Guangxi Zhuang Autonomous Region and Malipo County, Yunnan Province—areas along the border between China and Vietnam. Its presence in other regions of China remains uncertain. Since 2022, eight specimens of Bufo species have been collected from Dawuling Forestry Station, Xinyi City, Guangdong and three from Guangxi Yachang Orchid National Nature Reserve, Leye County, Guangxi (Fig. 1). The morphological characteristics of these specimens were examined and measured via a digital vernier caliper for preliminary species identification. To further confirm species delimitation, we used MEGA 11, raxmlGUI 1.3, and MrBayes 3.2.4 to construct phylogenetic trees. In addition, uncorrected p-distances were calculated in MEGA 11 based on mitochondrial 16S rRNA, COI, and ND2 gene sequences. Morphological comparisons revealed that these specimens matched the diagnostic characteristics of B. rubroventromaculatus (Table 2 and Fig. 2). These specimens had the following differences from the type specimens:head width larger than length; vomerine teeth present or absent; tympanum prominent; dorsal skin warts smooth with relatively few keratinized spines; snout-vent length larger than the type series in males. The phylogenetic analysis based on mitochondrial 16S rRNA, COI, and ND2 genes showed that all specimens of Bufo species clustered into a monophyletic lineage with a highly support rate (Fig. 1). The genetic distances between the populationfrom Xinyi City, Guangdong and the population fromLeye County, Guangxi, China were 0.36% (16S rRNA), 2.48% (COI), and 2.33% (ND2), respectively, while the genetic distance between these two populations and the Vietnamese population were 2.46% (Guangdong) and 0.86% (Guangxi) based on the ND2 gene, respectively (Appendices 1﹣3). Taking into account the results of morphological comparison and phylogenetic analysis, we confirmed that the collected specimens belonged to B. rubroventromaculatus. This represents a new record of this amphibian species in Guangdong. The discovery expands the known distribution of B. rubroventromaculatus from southeastern Yunnan and western Guangxi to northwestern Guangxi and western Guangdong. It provides significant insights into the phylogeography and biogeography of the genus Bufo. Moreover, this study refines the morphological description and biological information of B. rubroventromaculatus.

Abstract:In the field of zoology, blood research is important in many aspects such as animal health, disease treatment, and biodiversity conservation. The continuous advancement of molecular biology technologies, particularly the application of single-cell transcriptome sequencing, has greatly promoted blood research in zoology. This paper reviews the application of single-cell transcriptome sequencing in blood research in zoology and analyzes current challenges and future development trends, aiming to expand the application of this technology in animal science. This review shows that the application of this technology in blood research in zoology is limited at present. This technology is mainly applied in invertebrate insects and crustaceans and less in vertebrates. The research content mainly focuses on blood cell mapping and description and immune defense and response.

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