Abstract:[Objectives] DNA-based research methods have been widely applied in the study of species taxonomy and phylogeny, ecological genetics, and conservation biology, however, the comparative studies on DNA samplings are generally scarce. In this study, we aimed to compare the performance and applicability of different DNA sampling and extraction methods by using the captive-bred Chinese giant salamander (Andrias davidianus) as study objects. [Methods] DNA samples were obtained by four sampling methods including buccal swabbing, skin swabbing, shed skin sampling and tail venous blood sampling (Fig. 2). DNA was extracted by four different methods including Kit method, high salt method, phenol-chloroform method and magnetic bead method based on the most accessible skin swabbing samples. All the DNA qualities and concentrations were detected by gel electrophoresis and nucleic acid protein analyzer, and each of the sampling DNA was identified by PCR and sequencing of the mitochondrial COI fragments. In addition, the daily and feeding behaviors of the experimental animals were observed after samplings. [Results] Our results showed that the daily and feeding behaviors of A. davidianus didn’t change visibly, thus the four sampling methods could belong to the category of non-disruptive DNA sampling that has minimal impact on the fitness, behavior or welfare of the experimental animals. Among the four different sampling methods, the tail venous blood sampling performs optimally, followed by buccal, skin swabbing, and then shed skin sampling in terms of DNA quality and concentration (Fig. 3a, b). However, the DNA obtained by four different extraction methods based on skin swabbing samples were generally similar (Fig. 3c, d). Although different sampling methods performed differences in obtaining DNA, all the extracted DNA could be amplified of the target COI gene fragments (Fig. 3e, f). [Conclusion] By summarizing the advantages, disadvantages and precautions of different DNA sampling and extraction methods (Table 1), we suggest that the most appropriate sampling method should be selected according to the experimental purposes and specific conditions. In terms of extraction methods, however, considering the toxicity of reagents, the complexity of operation, time and economic costs, the kit method is what we recommended. This study could provide some guidance for non-disruptive sampling and relevant molecular ecology studies of A. davidianus and other rare and endangered animals in the future.

Abstract:[Objective] Siberian Roe Deer (Capreolus pygargus) is a kind of important wildanimal with edible value, meidicinal and ornamental value for scientific research. To understand the biological characteristics of Siberian Roe Deer fibroblasts, trachea, lung and ear tissues were collected for cell culture. [Methods] In this study, wild male and female Siberian Roe Deers were collected as experimental materials from Daqingshan, Inner Mongolia Autonomous Region, China. The primary fibroblasts of trachea, lung and ear were established by tissue block monolayer culture method. The cell adherence rate, survival rate before freezing and after resuscitation, growth curve of cells from different tissue sources were explored, and the karyotype and the G-band characteristics were analyzed. Unless otherwise noted, all analyses were performed in at least triplicate. Student’s t-test was uses to calculate difference using Graphpad (Version 5.0) [Results] The experimental results showed that spindle-shaped or triangular cells were newly grown along the tissue blocks in the trachea and ear tissues of male roe deer after 2﹣4 days of culture. After 8﹣10 days of culture, the convergence degree of cells in the flask reached 60%, and after 11﹣12 days of culture, the convergence degree of cells in the flask reached 80% (Fig. 1). The morphology and growth of fibroblasts from trachea, ear and lung of the femalewere basically the same as those of the male (Fig. 2). The most densely populated fibroblasts from the trachea, ears and lungs were observed at P20, P12 and P9, respectively (Fig. 3). The adherence rates of fibroblasts from the three tissues increased gradually with time, and grew rapidly within 6 - 12 h. After 24 h, the adherence rates of fibroblasts from the trachea and ears were over 95%. The adherence rate of P3﹣P9 lung fibroblasts varied greatly, and the adherence rate reached more than 90% when the culture time was 24 h (Table 1). There was almost no difference in the cell survival rate of airway fibroblasts in different generations before cryopreservation, and the survival rates were all above 85%. There was little difference in cell survival rate between ear and lung fibroblasts in different generations before cryopreservation, and the survival rate was above 90%. The recovery rate of fibroblasts after freezing gradually decreased with the increase of cell passage times (Table 2). The growth curves of fibroblasts from the trachea, ears and lungs show a typical “S” shape (Table 3, Fig. 4). Mycoplasma test results were negative (Fig. 5). The results of H.E staining showed that the three kinds of fibroblasts grew in a spiral shape, with light blue-purple nuclei, light red cytoplasm and that long spindle-shaped or triangular cellswere typical fibroblasts in morphology (Fig. 6). Karyotype and G-banding analysis showed that the number of chromosomes in male deer fibroblasts was 2n = 70 and Bs = 5, among which there were 34 pairs of autosomes, with the morphological type of 12 submetacentric + 22 submetacentric, one pair of sex chromosomes, with the morphological type of X being metacentric, Y being st and five B chromosomes (Fig. 7a, Fig. 8a, Table 4). The number of chromosomes in female fibroblastswas 2n = 70 and Bs = 8, among which there were 34 pairs of autosomes, with morphological type was 29 submetacentric + 5 subacrocentric, one pair of sex chromosome with morphological type of X being submetacentric, and eight B chromosomes (Fig. 7b, Fig. 8b, Table 5). [Conclusion] In this study, three tissue-derived fibroblast cell lines from male and female roe deer were successfully established. When cultured in vitro, they grew well and maintained the stability of genetic stability. The karyotypes and G-banding altas of male and female roe deer chromosomes were drawn. This study provides material and basic technical support for further related research in the future.

Abstract:[Objectives] Distychus maculates is an endemic fish in Turks River of the Yili River basin. In order to protect its resources, we carried out its artificial breeding to explore the embryonic development and post-embryonic development pattern. [Methods] In May 2021, artificial breeding of D. maculates was carried out, the embryonic and post-embryonic development were observed and photographed with a dissecting microscope, and the morphological characteristics of each developmental stage were recorded. Embryos and early larvae were randomly selected each time. The biological indexes of embryos and larvae were measured with Image pro plus 6.0 software. Photoshop CS6 was used for image processing and the SPSS 16.0 software was used for data processing. In this study, K = NT, where K is accumulated temperature (℃?h), N is the time required for development to a certain stage (h), and T is the average water temperature at this stage (℃). Descriptive statistics was presented as mean ± standard error. [Results] In the water temperature of 15.0﹣16.0 ℃, the embryo development of the fish was divided into 7 key stages: fertilization, cleavage, blastocyst, gastrula, neuroembryo, organogenesis and membrane emergence, and the membrane was brokenafter 2 969.830 ℃?h. The mature eggs were pale yellow with a diameter of 2.75 ± 0.24 mm (n = 20). After 35 min of fertilization, the largest perioocyte gap was 3.46 ± 0.16 mm (n = 20), and the volume of yolk accounted for about 3/5. The relationship between the length of larval fish (LT) and the days post hatching (D) was as follows: LT = 0.50D + 9.15 (R2 = 0.90); The relationship between the length of yolk-sac (LY) and the days of membrane emergence (D) was: LY =﹣0.11D2 + 7.61 (R2 = 0.76); The relationship between larval length (LT) and yolk-sac volume (LP) was as follows: LT =﹣2.35LP + 19.96 (R2 = 0.88); The relationship between yolk-sac total length (LY) and yolk-sac volume (LP) was as follows: LY = 0.49LP + 5.34 (R2 = 0.68); The relationship between yolk-sac volume (LP) and membrane extraction days (D) was as follows: LP = 0.002D2﹣0.24D + 4.67 (R2 = 0.98). The total length and volume of yolk sac decreased with the growth of body length. The yolk sac was consumed 15 days after membrane emergence. The whole length of larva increased with the days after membrane emergence. The whole length of larva increased significantly in yolk sac absorption stage. [Conclusion] By studying the artificial reproduction of D. maculates, we explored its characteristics of embryo and post-embryonic development, to enrich the data of its early life history, and provide theoretical data for the breeding of unique fish seedlings, which further lays a foundation for the protection and development of germplasm resources.

Abstract:[Objectives] Determining the elevational distribution of species diversity along altitudinal gradients is of critical importance in biodiversity and conservation research, the hump-shaped pattern of species diversity is the most common one. To understand the species diversity distribution pattern of Taibai Mountain, and to provide basic data and references for the study of elevational biodiversity gradient patterns. [Methods] We surveyed the diversity of small mammals along the elevational gradient on the northern slope of Taibai Mountain from June to September in 2020 and 2021. The elevation range of the study was 780﹣3 767.2 m, with a gradient of 200 m. Fifteen sampling points were set, with a total of 4 150 rat traps, 105 traps and 8 sample sites. Four diversity indices (Shannon-Wiener, Margalef, Simpson, Pielou) were used to compare the diversity discrepancy between different elevational forest types and used to reveal the diversity distribution pattern. All species diversity index calculation, cluster analysis was completed in SPSS 21.0 and R language. [Results] Our study surveyed totally 148 individuals representing 21 species that were belong to 12 genera, 7 families and 4 orders. The species diversity of Quercus variabilis and Q. aliena which in the middle altitude area was the highest, and that of Abies fargesii in high altitude area was the lowest. The elevational distribution pattern of species diversity of small mammals in the northern slope of Taibai Mountain Nature Reserve is a left-biased mid-peak pattern. The two non-parametric estimations of species richness Chao2 and Jackknife2 indicate that increased sampling intensity is still needed in Betula albosinensis. [Conclusion] The species diversity of small mammals in broad-leaved forest was generally higher than that in coniferous forest. This distribution pattern was mainly related to the composition of Oriental fauna and Palaearctic fauna, endemic species and non-endemic species of small and medium-sized mammals in the community. The mountain range-valley landform on the northern slope of Taibai Mountain provides favorable conditions for the diffusion and migration of northern and southern species. The climate and geographical characteristics in the valley were similar, forming a typical elevational vegetation zone, which made the distribution of species diversity of small mammals had similar characteristics.

Abstract:Brandt's Vole (Lasiopodomys brandtii) is one of the main rodent species in the typical steppe area of Inner Mongolia. The voles store food in the burrows in autumn to survive the winter when the vegetation is poor. [Objectives] In order to study the effects of increasing food on vole's winter survival rate. [Methods] In October 2004, we selected 2 sample plots (200 m × 100 m) in Baiyintuga, Abagaqi, Inner Mongolia, where is Brandt’s vole distributed. We defined 2 plots as increasing food plot and control plot. In the increasing food plot, we increasing wheat around all vole’s burrow clusters. Each burrow cluster was fed 500 g of wheat per day for two consecutive days for a total of 1 000 g. And do nothing in the control plot. The mark-recapture method is used to estimate the number of Brandt’s vole in 2 plots. The number of Brandt's voles was 310 and 318, respectively, and the results were used as the base for calculating their winter survival rate. In May 2005, we returned the 2 plots again to investigate the vole’s population number. Statistics using chi-square test. [Results] The winter survival rate of Brandt’s vole in the increasing food plot is 41.3%, which is significantly higher than the control plot’s winter survival rate of 24.2% (P < 0.01, n = 628, χ2 = 20.82) (Table 1). The winter survival rate of female and male Brandt’s vole in the increasing food plot is 45.4% and 37.3% respectively, which is significantly higher than the 25.8% and 22.6% winter survival rates of female and male Brandt’s voles in the control plot (P < 0.01, n = 311, χ2 = 13.07) (P < 0.01, n = 316, χ2 = 8.16) (Table 3). However, there is no significantly difference in winter survival rate between females and males in each plot (P > 0.05) (Table 2). [Conclusion] Increasing food will raise the winter survival rate of Brandt’s vole, which play an important role in population growth in the next year.

Abstract:[Objectives] Populations of long-distance migratory passerines have been declining continuously, and such decline has been especially striking in Rustic Bunting Emberiza rustica. Through a long-term monitoring and migration dynamics analysis of Rustic Bunting, this study aims to provide a scientific basis for more effective conservation actions. [Methods] We collected and analyzed the data on Rustic Buntings between 2001 and 2018 from 10 banding stations in Northeast China, including Gaofeng, Qingfeng, Maoershan, Xinqing and Dazhanhe in Heilongjiang Province, Hunchun and Jilin in Jilin Province, Liaoning bird research center in Dalian and Lvshun Laotieshan in Liaoning Province as well as Wuerqihan in Inner Mongolia. Mist-net was set nearby water on the edge of the forest where bird inhabits and checked every 30 minutes. The captured birds were banded and recorded before being released. In Gaofeng, Maoershan and Qingfeng banding station, the survey was conducted from 15th, March to 31th May in Spring and from 15th August to 20th November in Autumn. 70 mist-nets with 3m in height and 20m in length were set in different habitats in these three banding stations. We conducted statistic analysis on data from these three banding stations and used data from the rest of the banding stations for auxiliary analysis. The sex of Rustic Buntings is mainly distinguished by the color of the head, the color of the midline of the crown feathers, the width of the crown feathers (In males, the crown feathers are black in spring and the ear feathers are black while the base of the crown feathers is black in winter. Female's crown feathers and ear feathers are not black and the midline of the crown feathers are dark brown in winter). The age of Rustic Buntings is distinguished by the uniformity of the tail feathers-growth line, the degree of wear in tail feathers-tip, the width of the outer tail feather, the situation of molt in flight feathers and the color of iris. Birth to the end of the year is recorded as a juvenile. The following spring is recorded as sub-adult and the following autumn and beyond is recorded as an adult. [Results] A total of 184,181 Rustic Buntings were banded in our 18 years survey, including 88 571 in spring and 95 610 in autumn. The annual banding number fluctuated greatly and showed a rapid downward trend. The overwintering mortality rate of juvenile birds was 41.3% higher than that of adults. The data on the 106 recovered Rustic Buntings indicated that under natural conditions， Rustic Buntings enjoy a life span of up to 11 years, and can travel as far as 300 kilometers a day. Northeast China is an important migration channel for Rustic Buntings, with a relatively stable migration route. It is where Rustic Buntings would fly through on their way to the south of Tianjin for overwintering after breeding in northern Sweden. [Conclusion] The results showed that the Rustic Bunting population decreased rapidly in recent years, as the banding number decreased by more than 95% compared with the peak year, which deserves our greater attention. Habitat fragmentation and illegal hunting are two major threats to Rustic Buntings. It is suggested that efforts should be made to strengthen habitat protection, keep long-term ringing monitoring, and explore migration patterns of Rustic Bunting, so as to facilitate its population recovery.

Abstract:[Objectives] Amphibians and reptiles are important components of biodiversity and important groups of ecosystems. In this study, we investigated the diversity and altitudinal distribution characteristics of amphibians and reptiles in Yingjing Area of the Giant Panda National Park (102°19′﹣102°55′ E, 29°28′﹣29°56′ N). [Methods] From May to September 2021, we set up six observation sites in the Yingjing Area of Giant Panda National Park, and set up 33 effective line transects to investigate the diversity and elevation distribution of amphibians and reptiles, and supplemented the field survey results by interview survey and literature review (Appendix 1). We calculated the Shanoon-Winner’s diversity index, Pielou’s evenness index and ecological dominance index by Spass software (IBM SPSS Statistics 21.0). In addition, Pearson correlation analysis was conducted for these three indices in six study sites. Finally, we analyzed the correlation between the distribution of amphibians and reptiles and the elevation. The statistical test was set two-tailed, and the nominal significance level was set at P = 0.05. [Results] There are 67 species of amphibians and reptiles in the region, including 31 species of amphibians in 9 families of 2 orders and 36 species of reptiles in 9 families of 2 orders. All amphibians are Oriental species except Pelophylax nigromaculatus. There are 26 species of Middle Eastern Ocean reptiles, and only 9 species are widespread species (Appendix 2). By comparing the habitat types of amphibians in the region, we found the local amphibians were different in their adaptability to the environment (Table 1). The Shanoon-Winner’s diversity index, Pielou’s evenness index and ecological dominance index of the six observation sites showed similar change trends with Xinmiao > Paocaowan > Niba mountain > Yunwu mountain > Dashiba > Anjing (Table 2). [Conclusion] The abundance of amphibians decreased gradually with the increase of altitude gradient. Due to the limited number of reptiles, we did not find obvious altitude distribution characteristics (Fig. 3).

Abstract:[Objectives] During the fieldworks in April-May and July-August 2021, we collected 8 specimens of Amolops (Anura: Ranidae) from Gaoligong Mountain, Yunnan Province, China, which were identified as A. deng and A. chayuensis by comparison of Morphology and Phylogeny. This represents first records of these species in Yunnan Province. [Methods] In this study, the morphological characteristics of the specimens we collected were compared and analyzed with the holotype of A. deng and A. chayuensis, respectively. Based on 16S rRNA gene and COI gene fragments, we used Mrbayes-3.1.2 to construct bayesian tree for phylogenetic analysis. Based on 16S rRNA gene fragments, we used p-distance in MEGA 7 to calculate the genetic distance between individuals of Amolops. All of the new sequences from this study have been uploaded to the NCBI GenBank, the other sequence we used were obtained in NCBI GenBank database (Table 1). [Results] The morphological characteristics are consistent between the specimens collected in this study and holotype. A. deng has a medium body size (SVL 63.1 to 68.5 mm in adult female, n = 3, 49.3 to 52.9 mm in adult male, n = 3); head length is larger than the head width; tympanum is obvious, and taller than the length; three metacarpal tubercles; supratympanic fold is not obvious; dorsolateral fold obvious and connect with the crotch; tibiotarsal articulation beyond snout. A. chayuensis has a small body size (SVL 44.7 mm and 44.8 mm in male, n = 2); males with external subgular vocal sacs; supratympanic fold and dorsolateral fold obviously; arms and legs with brown bands. In addition, the bayesian analyses strongly support the specimens we collected forms a monophyletic clades respectively with each holotype specimens of A. chayuensis and A. deng from Tibet, China (Fig 3). Based on 16S rRNA, the genetic distance of A. deng was 0.8%﹣1.2% and A. chayuensis was 0.2% between the samples we collected and type locality (Table 3). [Conclusion] In summary, the two species of Amolops collected in Gaoligong Mountain in Yunnan are determined to be A. deng and A. chayuensis. Both A. deng and A. chayuensis are first record of Gongshan and Fugong counties of Yunnan province (Fig. 4). In this study, we further updated the elevation range of the distribution of A. chayuensis, from 1 385 m lower to 2 884 m higher than the previous record.

Abstract:[Objectives] The black Amur bream (Megalobrama terminalis) is an indigenous fish species, which has been identified to hold an important position in fishery production in the lower reaches of the Pearl River. This study aims to investigate the inter-annual variation on body length-weight relationship, Fulton’s condition factors (K) and age structure of M. terminalis population in the course of 2010-2018 in the downstream of Pearl River. [Methods] From 2010 to 2018, 1 785 specimens of M. terminalis were collected. Body mass and body length of all the samples were measured. In this study, we investigated the growth pattern of M. terminalis, basing on the body length, weight and age structure of M. terminalis. The power function of the body length and body weight relation equation: W = aLb was used to compare the variation trend of M. terminalis from 2010 to 2018. SPSS software (version 19.0) was used for data processing. We examined significant differences between allometric growth parameters b and 3 by t test. Descriptive statistics was presented as mean ± standard deviation (Mean ± SD). [Results] The results indicated that there was a decreasing trend in body length and proportion of individuals over 5+ years of M. terminalis population in Zhaoqing section of the Lower Pearl River from 2010 to 2018. Meanwhile, the body length-weight power function correlation condition factors (a) ranged from 0.011 to 0.004, and the value of allometric factor (b) ranged from 2.816 to 3.261 in the present study. M. terminalis population showed negative allometric growth in 2010, 2011 and 2014, while positive allometric growth in 2013, 2015-2018. The fatness of M. terminalis population presented a gradually increasing trend during 2010 to 2018. In addition, the proportion of daily average water temperature above 20 ℃ in ZhaoQing section of the Pearl River gradually increase, while the number of flood peaks decreased significantly and the distibution of flood peaks turned into intensively. Moreover, there was a positive relationship between allometric factor and water temperature and discharge, verifying the relevance between fish growth and habitat environment. [Conclusion] In general, high fishing pressure might be responsible to accelerate growth rate of M. terminalis population in the downstream of the Pearl River, causing negative allometric growth of M. terminalis population. Furthermore, implement of the fishery ban management and variation of water temperature in the lower reaches of the Pearl River might lead to the ascent in fatness of M. terminalis population.

Abstract:[Objectives] Amphioxus is a transitional animal from lower invertebrates to higher vertebrates, which plays an extremely important role in the research of origin and evolution of vertebrates. Currently, the family Branchiostomatidae contains three genera: Branchiostoma, Asymmetron and Epigonichthys. In August 2021, when conducted a comprehensive survey in Nanji Islands National Marine Nature Reserve, Zhejiang Province, the author discovered amphioxus which is a national second-class protected animal. [Methods] Compared the specimens (one male ZJ 01♂, one female ZJ 02♀) with other species of amphioxus by comparing their morphological characteristics and measurement data (Fig. 1, Table 2). Phylogenetic analysis was performed on the basis of COI sequences of the samples and other amphioxus to determine the phylogenetic location of the specimens. The Kimura2-Parameter model in MEGA11.0.1 was used to calculate the genetic distances between species of the amphioxus. [Results] We found the specimens were similar with Branchiostoma japonicum in morphological. Based on molecular evidence, the specimens were clustered with the B. japonicum (Fig. 2). Genetic distance estimation based on Kimura2-Parameter model showed that the distance between the specimens and B. japonicum was the closest, ranging from 0.006 to 0.013, the distance much lower than that between other species range from 0.171 to 0.269 (Table 3). [Conclusion] Combined with morphological and molecular biological methods, this species was identified as B. japonicum. According to relevant literatures, the author presumed that amphioxus in Nanji Island were the pelagic larvae of amphioxus which came from Taiwan Warm Current in the coastal waters of Fujian province. It is the first recorded species in Zhejiang Province, which will provide scientific data for the protection of amphioxus resources.

Abstract:Henan locates in the central of China and is characterized with transition area between Palearctic and Oriental regions in zoogeography. However, there is only few investigations of the third species-most abundant order Eulipotyphla, therefore limiting the study of zoogeography. In this study, we aimed to identify three shrews collected from Xinyang City, China, which would enrich the species diversity of Eulipotyphla in Henan Province, China. During October 2013, a shrew (voucher number IBE20131005) was collected from Baichong conservation station (31°36?54? N, 114°51?13? E) of Liankangshan National Nature Reserve, Xinxian, Xinyang, Henan Province, China; and during May 2018 two shrews (voucher number IBE2018100501 and IBE2018100502) were collected from Dahuwan (32°18?57? N, 113°48?5? E), Wujiadian Town, Shihe District, Xinyang, Henan Province, China. Based on the morphological characteristics (Fig.1), the three shrews could be assigned as species from genus Crocidura. In order to identify the taxonomic status, the morphological and skull variables of these shrews were measured and compared with topotype specimen (Table 1), and then phylogenetic relationship (Fig. 2) within genus Crocidura was analyzed using Cyt b sequences and K2P genetic distance was also estimated by using Cyt b sequences. The cranial length (CIL) of the specimen IBE2018100501 is 20.40 mm (Table 1). The cranial width (CB) is 9.62 mm, the orbital width (IOB) is 4.79 mm, the great width measured at anterior labial margins of 2nd upper molars (M2-M2) is 6.16 mm, the upper dentition length (UTRL) is 9.14 mm, and the lower dentition length (LMTR) is 8.13 mm. The genetic distance of K2P between Henan Province and other regions is 0.1%﹣1.2%. Both morphological (Fig. 1) and phylogenetic analyses (Fig. 2) suggested that the three shrews were Crocidura tanakae, a new record of mammals in Henan Province, China. This study also suggested the north-most distribution of C. tanakae was higher than previous record. The C. tanakae collected from the three locations of Mt. Dabieshan showed a very close phylogenetic relationship but further phylogenetic relationship with populations collected from Mt. Huangshan area located close to the south edge of Changjiang River, which suggested the river could be a natural barrier for the gene flows of this species. The specimens (voucher number IBE20131005, IBE2018100501 and IBE2018100502) were stored at the Institute of Biodiversity and Ecology, Zhengzhou University.

Abstract:The Red Deer (Cervus elaphus) plays an important role in ecosystem function in northeast China, but their populations had seriously suffered from habitat degradation and human disturbance. In this study, we investigated the population recovery and habitat status of red deer by means of line transects and pellet group counting methods in southern part of the Greater Khingan Mountains, Inner Mongolia. The results showed that after 10 years’ development, the population density increased significantly in Huanggangliang, Baiyin’aobao, Dalengshan, Saihanwula, Wulanba and Gaogesitai areas (P < 0.05). The total number of red deer in the study region was about 9 644.2 ± 1 378.6 ind, and the population density was 2.5 ± 0.9 ind/km2. This study provided solid baseline data for red deer conservation in the southern part of the Greater Khingan Mountains, and we further discussed the measures to promote a steady recovery of the red deer population according to habitat restoration and population management.

Abstract:Six bird species and subspecies were newly recorded during a bird survey in Medog, Southeast Tibet of China from October 2018 to October 2021, including Large Blue Flycatcher (Cyornis magnirostris), Mountain Scops-Owl (Otus spilocephalus), Golden-breasted Fulvetta nominate subspecies (Lioparus chrysotis chrysotis), Slender-billed Scimitar-Babbler intextus subspecies (Pomatorhinus superciliaris intextus), White-gorgeted Flycatcher nominate subspecies (Lioparus chrysotis chrysotis), and Buff-breasted Babbler garoense subspecies (Pellorneum tickelli garoense).

Abstract:This paper introduces the distribution of Elysia tomentosa K. R. Jensen, 1997 in Hainan Island for the first time. E. tomentosa belongs to Family Plakobranchidae, Superorder Sacoglossa, Class Gastropoda in the Phylum Mollusca. In September 2021, the sea slug was collected in Yunlong Bay, Wenchang city, Hainan Province. We designed the primers of 16S ribosomal RNA gene sequence, histone H3 gene (H3) sequence and tufA gene to identify what species it is and the algae in its living environment by polymerase chain reaction (PCR). We detected the photosynthetic fluorescence characteristics of E. tomentosa by chlorophyll fluorescence induction curve. A series of fluorescence parameters Fv/Fm, Y(II), Y(NO) and NPQ measured by the chlorophyll fluorescence measuring instrument Dual PAM-100 (Walz, Germany) were calculated by t-test with three samples and three repeat. Through sequence alignment of 16S rRNA and the relationship between photosynthetic characteristics and evolution, a phylogenetic tree was constructed by maximum likelihood (ML) method. The comparison of 16S rRNA sequence and H3 gene sequence identified this species as E. tomentosa by molecular method. The source of the symbiotic chloroplasts of this species was Caulerpa racemosa (Fig. 3). The photosynthetic activity of E. tomentosa could maintain for about one month and the photosynthetic capacity of E. tomentosa continue to decrease (Fig. 2). The evolutionary tree showed that the evolutionary status of E. tomentosa in the some species (Fig. 4). This is another species of photosynthetic mollusk discovered by our research group in Hainan Province, which further enrich our understanding of its distribution in China and also provides new materials for studying the evolution of photosynthesis and endosymbiosis mechanism.

Abstract:The Crocodile Lizard (Shinisaurus crocodilurus) is the first-class protected wild animal. In recent years, crocodile lizards in the Beilou Station in Guangxi Daguishan Crocodile Lizard National Nature Reserve in China have been plagued by diseases, but the reasons are unknown. To investigate whether the occurrence of these diseases was related to the pollution of heavy metals and pesticides in food, this study detected heavy metals and pesticide residues in their main food using inductively coupled plasma mass spectrometry and chromatography mass spectrometry. Results showed that heavy metals were lower in the crocodile lizards compared with their food (earthworms), while pesticide residues were not detected in crocodile lizards and their food. These results indicate that it is less likely that heavy metals and pesticides accumulate in the crocodile lizards through biomagnification. Therefore, environmental pollutants such as heavy metals and pesticides have little effect on the occurrence of crocodile lizard disease. This study provides a references for the artificial rescue and breeding of crocodile lizards, which is benefit to the protection Crocodile Lizards.

Abstract:Pungitius sticklebacks (Gasterosteidae) widely distributed across the Northern Hemisphere. Phylogenetics relationships and taxonomic validity of different species and/or populations remain contentious due to convergent evolution and frequent hybridization among the Pungitius sticklebacks. Here, we briefly reviewed the taxonomic history and distribution of the genus Pungitius, and verified the phylogenetic relationships and phylogeography of the genus from around the world, especially from China. This paper further confirmed the findings of Wang et al. (2021), i.e., there may be four species of Pungitius sticklebacks in China, including P. sinensis, P. kaibarae, P. bussei, and P. stenurus, among which the P. sinensis has two different lineages. We also suggested conservation and management authorities dealing with sticklebacks should resort to molecular species identification when planning translocations in order to more scientific and efficient artificial release and protection of sticklebacks.

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