Abstract:Scales in squamate reptiles are the main features that distinguish them from the other amniotes. The ultrastructure of scales in squamate reptiles was an adaptation to the local environment. We used scanning electron microscope to observe the microornamentation and scale sensilla on heads, middle dorsal and middle ventral parts of Takydromus septentrionalis, Dopasia harti and Elaphe carinata. Our results showed that there were interspecies and intraspecific differences in the microornamentation and scale sensilla of the examined species: 1) oberhautchen of T. septentrionalis and E. carinata was consisted of ?at and strap-shaped cells, but oberhautchen of O. harti was formed by polygonal cells; 2) the lenticular scale sensilla existed on the chin-shields of O. harti and E. carinata, while there was no lenticular scale sensilla on the chin-shields of T. septentrionalis; 3) the posterior margin denticulations were present on the oberhautchen of T. septentrionalis and E. carinata, however, the denticulations of E. carinata were wider and longer than in T. septentrionalis; 4) the oberhautchen cells appeared erect ridge on circumocular scales, while flat and strap-shaped cells were found on the scales of other parts in E. carinata. Simultaneously, a total of 17 families and 99 species of dorsal scale microornamentation data and 8 families and 25 species of receptor data were collected and ancestral reconstructions were performed on terrestrial squamate microornamentation and scale sensilla. The results showed that the narrow strap-shaped cells mainly existed in the Lacertide, Colubridae and Scincidae, while the polygonal cells existed in the Agamidae, Anguidae, Boidae and Viperidae. The ancestral morphology of scale sensilla was lenticular-like and lenticular, furthermore, the lenticular structure has evolved repeatedly. In addition, we speculated that the microornamentations and scale sensilla of the O. harti were closer to those snakes. Therefore, it was deduced that the morphological features of the microornamentations and scale sensillum of squamates contributed to their ecological adaptation to preferential microhabitats.

Abstract:Aggressiveness is one important dimension of animal personality, measuring an animal's tendency to express provoking and attacking actively toward other individuals. Animal aggressiveness has crucial connection with the social structure and hierarchy. In this study, the focal sampling and all-occurrence recording were used to sample the conflict behavior of captive Forest Musk Deer (Moschus berezovskii) in the Maerkang Musk Deer Breed Center of Sichuan from June 1, 2018 to July 31, 2018, and the aggressiveness personality (aggressiveness index) and hierarchy index of individuals were calculated respectively. The aggressiveness and hierarchy pattern of captive forest musk deer were analyzed (Kruskal-Wallis and Mann-Whitney U), and the influencing factors and interrelationships between aggressiveness and hierarchy were also explored (Spearman relationship test). The results showed that there existed significant difference in aggressiveness between gender, in which the male was significantly more aggressive (0.45 ± 0.09, n = 22) than the female (0.22 ± 0.06, n = 30) (P < 0.05). Age and group density had no significant effect on aggressiveness (P > 0.05) of captive forest musk deer, and age and gender had no significant effect on hierarchy (P > 0.05). In addition, there was a significant positive correlation between the aggressive and hierarchy. The more aggressive the individual was, the higher the rank was in a group (r = 0.727, P < 0.05). It was speculated that this was related to the hierarchy construction and resource competition in the musk deer group.

Abstract:In order to explore the community status of small mammals in Dongting Lake area after the Three Gorges Project, the small mammals in the marshland habitat of Dongting Lake area were investigated. From 2010 to 2018, the diversity of small animal communities in the marshland habitat of Dongting Lake area was investigated by snap traps, and the dominance concentration index, community diversity index and evenness index were calculated. The diversity index was compared by t value method, and the capture rate and rodent species composition of each year and habitat were tested by Chi-square test. The results of investigation as follow. From 2010 to 2018, a total of 34 116 traps were placed, 2 129 samples of distinguishable species were captured. Rodentia and Soricomorpha were the main species of small mammals captured in Dongting Lake beach habitat, including 7 species, Microtus fortis, Apodemus agrarius, Rattus norvegicus, Micromys minutes, Niviventer confucianus, R. losea and Suncus murinus. The total capture rate is 6.36%, among them, the capture rate of Microtus fortis and A. agrarius was higher, was 1.83% and 3.94% respectively (Table 1). From the composition of small mammals species in Dongting Lake beach, the dominant species are A. agrarius and Microtus fortis, and their proportion in the community is 63.18% and 29.26% respectively (Table 2). Among the five types of habitats investigated, the diversity index (1.077 2) and evenness index (0.666 2) of the small animal community in the habitats mainly composed of Carex sp. + Miscanthus sp. + Populus deltoides were the largest. It was followed by habitats dominated by Carex sp. (diversity index 0.927 5 and evenness index 0.572 3); habitats dominated by Miscanthus sacchariflorus (diversity index 0.885 6 and evenness index 0.550 2); and habitats dominated by Carex sp. + Miscanthus sp. (diversity index 0.775 6 and evenness index 0.481 9). The diversity index (0.323 6) and evenness index (0.180 6) of the habitat dominated by P. deltoides was the lowest, but the dominance concentration index was the highest (0.881 2) (Table 6). The results showed that with the operation of the Three Gorges Project, the vegetation succession in Dongting Lake area changed, which caused the dynamic change of the small mammal community. On the marshland of Dongting Lake, the more complex the habitat, the higher the diversity of small mammal species. In the single habitat, the greater the interference of human activities, the lower the diversity of small animal species. At present, the A. agrarius has become the first dominant rodent species in the marshland habitat of Dongting Lake.

Abstract:Monitoring of mammals and birds is an important part of biodiversity conservation. From April 2017 to May 2018, we installed 36 infrared-triggered cameras in Laoxiancheng National Nature Reserve of Shaanxi Province in 36 1 km x 1 km grids, and evaluated the species richness, relative abundance and photographic rate of mammals and birds. With a survey effort of 7 835 camera-days, we obtained 1 915 independent photographs and identified 32 mammals and birds belonging to 8 orders and 18 families. Among the recorder species, two were Endangered, four were Vulnerable, and three were Near Threatened on the IUCN Red List. The species with the highest relative abundance index (RAI) value in the reserve is Takin (Budorcas taxicolor) (IRA = 36.81) (Appendix 1). The species photographic rate of summer and autumn is higher than spring and winter (Fig. 3). There was no significant difference in the number of species and species diversity index at different seasons. There was also no significant difference in photographic rate, number of photographed species, and species diversity index at altitude. But there were significant differences in the photographic rate of the Takin at altitude (Table 1). Principal components analysis (PCA) showed that there had the correlation between the camera and the photographed species (Fig. 6 and Fig. 7), which was related to the habitat around the camera. In addition to providing a preliminary evaluation of mammal and species in the Laoxiancheng Nature Reserve, we also assessed the factors affected the distribution of species. This is the first study to investigate direct and quantitative data on species occurrences in this reserve. Our study provides important basis for future local wildlife research and conservation management.

Abstract:In July 2019, a bat was captured at Shihua Cave in Danzhou of Hainan Island and identified as Eonycteris spelaea by morphological measurements, skull characteristics and analysis on mitochondrial Cyt b gene. It is medium-sized bat with a forearm of 69.6 mm and without a claw on the second finger of the forelimb (Table 1). A short tail of 12.9 mm is prominent behind the femoral membrane and the bat has well-developed anal gland (Table 1 and Fig. 1). The greatest length of skull (GTL) is 35.2 mm and the rostral length (RL) is 11.4 mm (Table 1). The phylogenetic relationships of E. spelaea with its close species inferred from Cyt b gene sequences display that the Hainan bat and E. spelaea cluster a branch in high bootstrap value (Fig. 2). This is the first record of this species in Hainan Island. In the present study, morphological characters, the skull characters and the phylogenetic tree based on Cyt b gene sequences were represented detailedly. Comparisons were also performed between our findings and published data about the species recorded in other areas.

Abstract:In January 2018, when investigating chiropteran diversity Wuyi Mountain in Jiangxi province, a male vespertilionid bat was captured by using harp trap. We identified this bat through morphological examination and phylogenetic inference. It is medium-sized bat with a forearm of 38.29 mm (Table 1) and the black-brown fur with white tip. Its tragus is triangular. Ears are nearly square with transverse ridges and connect in the forehead without a prominent projecting lobe (Fig. 1a). Besides, skull rises evenly from the rostral to the end, the vertex of skull is not prominent (Fig. 1c). The zygomatic arch is slender, while both sagittal and lambdoidal crests are weak. Its dental formula is 2.1.2.3/3.1.2.3 = 34 (Fig. 1b, c). According to its external, skull characteristics and phylogenetic evidences using partial Cyt b and ND1 gene, it is identified as Barbastella leucomelas which representing a new record species in Jiangxi Province, China.

Abstract:Amphibian resources are abundant and unique in China, and more new amphibian species were found through molecular techniques. A Microhyla species was found during a field survey in Zhejiang Jiulongshan National Nature Reserve in April of 2017 and 2018. They were identified based on morphological comparison, BLAST comparison and phylogenetic analysis of mitochondrial 16S rRNA gene. Thirty-one adult (25♂♂ and 6♀♀) were captured and measured for snout-vent length (SVL), head length (HL), head width (HW), snout length (SL), internasal distance (IND), internorbital distance (IOD), diameter of eye (DE), length of lower arm and hand (LLAH), total length of leg (TLL) and foot length (FL) to examine sexual dimorphism of this species. Corresponding data were processed by one-way ANOVA, linear regression and principal component analysis. Our results showed that: (1) the species was identified as M. beilunensis and this sample locality is a new geographic distribution of M. beilunensis (Figure 1); (2) M. beilunensis displays female-biased sexual size dimorphism (Table 3); (2) other nine morphological traits were positively related with SVL in both sexes of M. beilunensis (Figure 3); (3) the mean value of LLAH in males was larger than female after removing the effect of SVL (Table 3); (4) a principal component analysis resolved two components (eigenvalues 1) from nine SVL-adjusted morphological traits, accounting for 52.47%% of variation in the original data (Table 4). The first component had high positive loading for HW and LLAH, and morphological traits between male and female differed significantly in their scores on the first axe (Table 4), indicating that HW and LLAH could distinguish females and males.

Abstract:The checklist of extant, native reptiles of Zhejiang Province, China is updated based on Fauna of Zhejiang Amphibia & Reptilia in 1990 and newly published data to October 2019, along with its fauna being analyzed. In total, 89 reptile species and one subspecies are recorded in Zhejiang, belonging to 54 genera, 18 families and three orders (Table 1, Appendix). Among these reptiles, there are 77 species and one subspecies of Squamata (belonging to 43 genera and 12 families), 11 species of Testudines (belonging to 10 genera and five families) and one species of Crocodylia (in one genus and one family) (Table 2). Among them, twenty species are endemic to China; two species, five species and 16 species are listed as the national first-grade, national second-grade and provincial key protected species, respectively. Furthermore, seven species are listed by CITES appendices I; six species by CITES appendices II; two species by CITES appendices III, respectively. According to the IUCN Red List of Threatened Species (IUCN-RL), two species and six species are listed as critically endangered (CR) and endangered (EN), respectively. Ten species and 11 species are listed in the Red List of China’s Vertebrates (RLCV) as critically endangered (CR) and endangered (EN) (Appendix 1). By realm, oriental species are dominant (69 species), and the others are widespread species (18 species) and palaearctic species (two species) (Table 3). By ecotype, terrestrial species are dominant (61 species) (Table 4). The results can be used as basic data for the systematics, phylogeography and ecology of reptile. This study has an important guiding significance to protect and monitor the reptiles in Zhejiang Province, China.

Abstract:To study the species composition and morphological characteristics of Siphonosoma in Zhaoshu Island, an onsite investigation was conducted in the tidal flat in Zhaoshu Island in Summer 2018. The environmental adaptability of Siphonosoma in tidal flat were analyzed based on the sediment characteristic and light microscopic observation of tissue biopsies. Three sampling section were selected in the tidal flat to collect and identify the macrobenthos qualitatively and quantitatively, and the sand grain size composition in gut and slices of gut were determined. The results indicated that, There are three species of Siphonosoma, and they are S. rotumanum, S. cumanense and S. funafuti, the species composition was about 4︰2︰1. The three species lived in the same environment and disturbed the same level of 10﹣15 cm. The gut of Siphonosoma was filled with sand, and the small organic particles, diatoms, copepods and plant fibers were found in gut. The large particles in down-gut were more than that in the upward-gut, and the small paticles in down-gut was less than that in the upward-gut. The result indicated that the gut of Siphonosoma has grain refinement effects to sediment particles. The cuticle of introvert retractor muscle and body wall were 59.08 μm and 231.92 μm, respectively. The introvert retractor muscle consisted of 9 000﹣9 500 muscle fibers, and there were some mastoid in the skin. The thicker cuticle and strong introvert retractor muscle were two adaptions of Siphonosoma to the coral reef sediment environment. The numbers of folds and muscle fibers in down-gut were less than that in the upward-gut. This may be correlated to the low content of organic matter in the coral reef sand.

Abstract:HTR8/SVneo cells are an important component of the placenta to establish blood circulation. Peroxisome proliferator-activated receptor alpha (PPARα) is a key factor regulating lipid metabolism. This study aims to clarify the influence of PPARα on human trophoblast cell function. The constructed PPARα expression vector and PPARα small interfering RNA were transfected into HTR8 / SVneo cells, respectively, to detect changes in cell function. EdU method and MTT method were used to detect cell proliferation, flow cytometry was used to detect cell apoptosis, and transwell chamber method was used to detect cell migration and infiltration capacity. The number of samples in each group is not less than three, and the experiment is repeated three times. The experimental data were statistically analyzed using GraphPad Prism 6 software. Data were processed using t test and analysis of variance. P < 0.05 was considered statistically significant. The results showed that PPARα overexpression inhibited the proliferation, migration and infiltration of trophoblast cells and promoted apoptosis (P < 0.05, Fig. 2a, 3a, c, 4a, b, 5a, b, 6a, b); knockdown of PPARα promoted cell proliferation, migration and infiltration, and inhibited apoptosis (P < 0.05, Fig. 2b, 3b, d. 4c, d, 5c, d, 6c, d). The level of PPARα expression is negatively correlated with its effect on cell growth and migration.

Abstract:This experiment used Grimelius argyrophil staining to observe and measure the effect of cyclic starvation on the morphological function and distribution density of argyrophilic cells in the digestive tract of rats (Rattus norvegicus). Rats were starved for one day after being fed for one day for a period of 14 days. SPSS 25.0 was used to conduct one-way ANOVA on argyrophilic cells in each part of the digestive tract in the experimental group and the control group. Moreover, a statistical analysis was conducted on the distribution density of the argyrophilic cells in the same part of the digestive tract by using the independent sample T test. The results showed that the argyrophilic cells were distributed in all segments of the digestive tract except for the esophagus in both groups. Morphologically, the argyrophilic cells in control group were mainly ellipse and cone-shaped (Table 1), while those in the experimental group were mainly cone-shaped (Table 1). The distribution density of argyrophilic cells in gastrointestinal tract of the two groups was the highest in stomach (Table 2). The density of arginophilic cells in the stomach in the control group was 44.95 ± 7.87, and that in the experimental group was 42.35 ± 10.95. Low density of arginophilic cells was found in the jejunum, cecum, ileum and rectum of control group (Table 2) and that in the experimental group was found from jejunum to rectum (Table 2). The distribution density of argyrophilic cells in in the cardia and pylorus of the experimental group was very significantly lower than in the control group (P < 0.01), and that in the colon was significantly lower than that in the control group (P < 0.05). These results indicate that cyclic starvation will significantly change the morphology of argyrophilic cells and reduce the distribution density of the cells in the digestive tract, which may be related to the changes in the function of the endocrine cells.

Abstract:The nutrition source plays an important role in the growth and development of animals. In this study, the effects of glucose and vitamin C (Vc) on the growth and activity of three enzymes (malate dehydrogenase: MDH, lactate dehydrogenase: LDH, amylase: AMS) of tadpoles of Rana zhenhaiensis were studied using the static water immersion method. Healthy and uniform-size sibling tadpoles of R. zhenhaiensis with developmental stages of Gosner 29 and 30 were selected for testing. For glucose treatment, three experimental concentrations were chosen: 0.5, 1.0 and 2.0 g/L, respectively; and for Vc treatment, three experimental concentration were chosen: 10.0, 20.0 and 30.0 mg/L, respectively. Treatment with dechlorinated tap water was set up for control. Each treatment was replicated three times, with 10 objective tadpoles. The experimental period was lasted for tadpole metamorphosis until forelimb emerged (Gosner 42). The results showed that survival rates among different glucose treatments were not significantly different (P > 0.05). Metamorphic time (P > 0.05) was significantly different among treatments, with the shortest period in the 0.5 g/L (43.0 ± 4.0 d) and 1.0 g/L (43.0 ± 3.4 d) group, and the longest period in the 2.0 g/L (46.2 ± 5.4 d) treatment. Both body mass and total body length of metamorphosis were found to be the largest in the 0.5 g/L treatment group. Body mass (P < 0.01) of metamorphosis among different treatments was significantly different, but total body length (P > 0.05) was not significantly different. Rate of weight growth was found to be the highest in the 0.5 g/L treatment (9.67 ± 1.71 mg/d) and the lowest in the control group (7.54 ± 1.22 mg/d), and the differences were significant (P < 0.05) (Fig. 2). For Vc treatments, like those in the glucose experimental treatments, survival rates among different treatments were not significantly different (P > 0.05). Developmental stage of each treatment was more advanced than that of the control when examined 1 week later, as well as on day 14, 21 and 28, respectively. Metamorphic time (all treatments were similar to 43.0 d, P > 0.05) was not significantly different among all treatments. Total body length and body mass of metamorphosis in control treatment were smaller than those of 20.0 mg/L and 30.0 mg/L treatment groups, but total body length (P > 0.05) and body mass (P > 0.05) was not significantly different among the treatments during the experimental period. Rate of weight growth was found to be the highest in both 20.0 mg/L and 30.0 mg/L treatments and the lowest in the 10.0 mg/L treatment, but there was no significant difference (P > 0.05) (Fig. 3). Both glucose and Vc could impact enzyme activity of R. zhenhaiensis tadpoles. The activity of MDH increased with the increase of experimental concentrations of glucose and Vc. The activities of LDH and AMS were highest in 1.0 g/L of glucose group and 10.0 mg/L of Vc group, respectively (Table 1). These findings suggest that 10.0 mg/L of Vc or 1.0 g/L of glucose should be the optimum exogenous concentrations that could promote the growth performance and enzymatic activity of R. zhenhaiensis tadpoles. These data might provide some valuable information for R. zhenhaiensis breeding in the future.

Abstract:Currently, the tree shrew has been widely used as an alternative laboratory animal in biomedical researches. Our previous study showed that Helicobacter spp. was the most distributed bacteria in gastro- intestinal of tree shrew, however, the species of this genus were still unknown. To find out the characteristics of Helicobacter spp. species carried in laboratory tree shrew, we performed the current study to provide useful information for future investigation. Seventy-two tree shrew fecal samples were collected, and processed for DNA extraction, nested PCR to amplify specific gene fragments with the genus specific primers. The species- specific primers including those for Helicobacter hepaticus, H. muridarum, H. bilis, H. rodentium, Flexispira rappini, H. suncus and H. typhlonius were used to amplify species-specific fragments from genus-positive samples. The samples negative for species-specific primers but positive for genus primer were sent for sequencing. BLAST and alignment methods were used to confirm their Helicobacter species. The results showed that eighteen feces were positive for Helicobacter, and the positive rate was 25%, among which eight strains were H. typhlonius, six were H. bilis, five were H. cinaedi, two were H. felis and one was H. macacae (Fig. 1). Four cases were mixed carrying both H. typhlonius and H. bilis. The carried Helicobacter genus and species had no statistical significance in tree shrew among different gender and age groups (P > 0.05) (Table 2). The highly carried rate of Helicobacter spp., mainly H. typhlonius, H. bilis and H. cinaedi, was found in laboratory tree shrew with different genders and ages.

Abstract:Silvery pomfret (Pampus argenteus) is a kind of valuable economic fish species and the intestinal gas bubble accumulation is an important factor that hinders its popularization. In order to evaluate the changes of the bacterial communities in Silvery pomfret’s intestine upon gas bubble accumulation occurrence, 15 diseased fish were chosen as the diseased group and 15 healthy fish as the healthy group. The method of high-throughput sequencing of 16S rDNA gene’s V3-V4 region was used to determine diversity and composition of bacterial communities in each of these samples. The comparison results of the Alpha diversity index showed that there was no significant difference in the Simpson index and the Shannon index between these two groups (P > 0.05), but the Chao1 estimator and the ACE estimator in the diseased group are significantly lower than those of the healthy group (P < 0.05) (Table 1). The analysis of the microbiota structure showed that the dominant bacterial groups were similar in the diseased and control groups, at the phylum level being Proteobacteria, and at the genus level being Sphingobium, Sphingomonas, Cupriavidus, Novo sphingobium and Acinetobacter (Table 2). From the above results, it can be concluded that the intestinal gas bubble accumulation reduces the richness and evenness of intestinal flora of Silvery pomfret, but does not affect its diversity. In addition, the two groups of samples were analyzed by LEfSe and the results showed that the relative abundance of aerobic bacteria such as agrobacterium bacteria, thermophilic bacteria, cyanobacteria, rhizobial bacteria in the diseased group was significantly higher than that of the healthy group, while the relative abundance of acinetobacter and microacidic bacteria was significantly lower than that of the healthy group (Fig. 3 and Fig. 4 ). From these results, it can be concluded that the intestinal gas bubble accumulation may cause intestinal flora disorder.

Abstract:Food enrichment experiments (wood-wool, sack and bamboo tube) were carried out on 8 Chimpanzees (Pan troglodytes) fed at Shanghai Zoo, and the orthogonal test was used to design the test factors and levels. We recorded feeding behavior using all-occurrence recording method before and after food enrichment. The results showed that the order of the factor on feeding behavior was wood-wool > sack > bamboo tube. The influence of different food enrichments on feeding behavior of adult male was not significant, while all the adult females decreased. The proportion of feeding behavior of young individuals decreased after enrichment. There was no significant effect on adult male and young individual in the three ways of enrichment. The optimal combination of enrichment for four adult female chimpanzees was wood-wool and sack (with or without bamboo tube), and the optimal combination for one sub adult female chimpanzee was wood-wool (with or without bamboo tube and sack). The effect of wood -wool on adult female's feeding behavior was very significant (P < 0.01), and the influence of bamboo tube on each individual’s behavior was not significant (P > 0.05), and the effect of sack on some individuals was not significant (P > 0.05) (Table 6).

Abstract:The complete blood count (CBC) of the South China tiger was analyzed by automated hematology analyzer and manual microscope method respectively in this study. By comparing the results obtained from the two methods, a suitable method for CBC of South China tiger was explored. Forty blood samples from South China tiger were collected and examined. When data detected by the two methods were normally distributed, paired sample T-test was used to analyze the difference, otherwise, Wilcoxon signed rank test was used. Correlation analysis was performed by Spearman analysis and the statistical significance was judged when P value was less than 0.05. The results showed that there was no significant difference for the red blood cells counts (P > 0.05), white blood cells counts (P > 0.05), neutrophil ratio (P > 0.05), eosinophil ratio (P > 0.05) between the two methods. And there were significant differences in the lymphocyte ratio (P < 0.05), monocyte ratio (P < 0.05) and basophil ratio (P < 0.05). The two sets of data were well correlated in red blood cells counts (r = 0.915), white blood cells counts (r = 0.832), lymphocyte ratio (r = 0.832), and neutrophil ratio (r = 0.481), while the correlation coefficients of monocyte ratio (r = 0.283), eosinophil ratio (r = 0.309) and basophil ratio (r = 0.146) were lower than 0.4, and P value was greater than or equal to 0.05, showing weak or extremely weak correlation. Thus, the automatic hematology analyzer can be used to accurately detect the count of both red blood cells and white blood cells of South China tiger, which has the advantages of accuracy, rapidity and reliability. However, this method is not suitable for the classification of white blood cells. If necessary, the accuracy of the results can be guaranteed by combining microscope and machine method. This study provides a reference for the rapid and accurate analysis of blood cells in South China tiger.

Abstract:On November 16 and 18, 2019 respectively, a single bird of the Yellow-vented Bulbul (Pycnonotus goiavier) was found at the Xishuangbanna Tropic Botanic Garden of the Chinese Academy of Sciences, in Mengla of Yunnan Province. It might be the first record of the Yellow-vented Bulbul in China. The bird was observed to be mixed in a flock of the Red-whiskered Bulbul (P. jocosus) and feeding on the fruits of Waringin (Ficus benjamina).

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