Abstract:From April to June 2019, the fixed-point observation method and sample method were used to investigate the nest-site selection of Bar-headed Goose (Anser indicus) in the Yanchi Bay National Nature Reserve of Gansu Province. A total of 332 Bar-headed Goose nests were found, including three types of nest sites: haystack nests, ground nests, and bare rock cliff nests (Fig. 2). All nests were distributed on haystacks in shallow water swamps, on islands in the lake, and on nearby cliffs (Fig. 1). The average densities of the bar-headed geese in the three nest habitats were 0.18 /m2 (n = 204), 0.59 /m2 (n = 108), and 0.06 /m2 (n = 20). Bar-headed Geese favored to choose a nest in a small island in the center of the lake, followed by shallow water swamps and cliffs (Table 1). The average clutch sizes of bar-headed geese in the three nesting habitats were 4.74 ± 2.68 (n = 204), 4.23 ± 1.94 (n = 108) and 3.13 ± 0.64 (n = 20), with the hatching success rates of 48.65% (n = 199), 45.27% (n = 148) and 24.00% (n = 25), and reproduction success rates of 66.67% (n = 42), 74.28% (n = 35) and 36.36% (n = 11) (Table 2). Independent sample t-test and Mann-Whitney U test were used for nest site quadrats and the same number of control quadrats in each camp nest habitat. We found that in shallow water marsh habitats, three indicators of haystack density, haystack cover and water depth exist. Very significant difference (P < 0.01), and there was significant difference in water area (P < 0.05). In the lake core island habitat, there are extremely significant differences in the six indicators of vegetation density, vegetation height, maximum vegetation height, vegetation coverage, bare land ratio, and hay ratio (P < 0.01); in the cliff habitat, the slope direction and distance There are extremely significant differences between the three indicators of grass beach distance and water distance (P < 0.01) (Table 3﹣5). The principal component analysis showed that the main environmental factors affecting the selection of nesting sites for bar-headed geese in shallow marsh habitats were: water source factor, hidden factor, disturbance factor, and food factor; the major environmental factors affecting nest site selection in the core island habitat of Huxin Island were: Hidden factors, food factors, interference factors; the main environmental factors affecting nest site selection in cliff habitats are: terrain factors, food and water sources factors, and interference factors (Table 6﹣8). Studies have shown that the main environmental factors of nest site selection are different in different nest habitats of Bar-headed Geese, which mainly depend on the characteristics of habitats and surrounding environmental factors.

Abstract:The species of the subfamily Sciurinae are widely distributed in China, which play an important role in the dispersal of plant seeds and are considered an important animal group to maintain the ecological balance in forest ecosystems. From November 2016 to November 2017, we used line transect and camera-trapping methods to survey the Sciurinae species in Mentougou District, Beijing. Thirty-three line transects and 72 camera-trapping stations were established in total. The length of all line transects was 205 km and the total camera-trapping efforts accumulated to 5 059 camera-days. Four Sciurinae species were identified, among which Sciurus vulgaris is a new record for Mentougou District, Beijing, and Sciurotamias davidianus is a species endemic to China. S. davidianus has the highest relative abundance, followed by Sciurus vulgaris and Tamias sibiricus, and Tamiops swinhoei the lowest. The four squirrel species occur sympatrically in some areas, and competition among species is likely reduced due to the difference in their activity time. This research elucidated the distribution and abundance of Sciurinae species in Mentougou District of Beijing, providing basic information for their subsequent monitoring and protection management.

Abstract:Understanding the activity patterns of Plateau Zokor (Myospalax baileyi) during the breeding season has a great importance to their survival strategies and ecological control for zokor damage. This study used the radio tracking technology to monitor the activity patterns of plateau zokor during the breeding period from March to May in 2019 in Maqu County, Gansu Province and analyzed the correlation between the activity pattern and the soil relative moisture as well as soil temperature at the depth with 0﹣10 cm. The results showed that: 1) the plateau zokors exhibited three peaks of daily activity pattern during the breeding period. These peak time appeared at 08:00, 14:00, and 20:00, respectively. (Fig. 1). 2) the daily activity difference index (θ) indicated that the activity intensity of plateau zokors varies significantly with uneven distribution during the breeding period. The diurnal index (λ) indicated that plateau zokor had more activities in daytime during the breeding period. The daily activity time index (AT) indicated that the total daily activity duration of the plateau zokor was about 8 hours (Table 2). 3) Correlation analysis showed that the activity intensity of plateau zokor in breeding period was positively correlated with soil temperature (0﹣10 cm) (P < 0.05) (Fig. 4), but not correlated with soil relative moisture (0﹣10 cm) (P > 0.05) (Fig. 5). Our research indicated that the activity intensity of plateau zokor was influenced by the soil temperature at depth between 0 and 10 cm.

Abstract:Tylototriton kweichowensis is an endemic species in China and was listed in Grade Ⅱ of National protected wildlife. It has important guiding significance for environmental changes. From September 2018 to October 2019, using field observations were made on the habitats, morphological characteristics, and reproductive behavior of T. kweichowensis in the demonstration area for comprehensive management of rocky desertification in Salaxi, Bijie City, Guizhou Province. The results show that, T. kweichowensis lives in mountain ponds, springs, reservoirs, and temporary puddles. As shown in Table 1, the total length, tail length and weight of females are higher than that of males. T. kweichowensis went out after a thunderstorm on April 18. The species began to breed on April 29, and finished their breeding at the latest on August the reproductive peak was May-June. As shown in Fig. 3 and Fig. 4, the operational sex ratio of T. kweichowensis in breeding period is bias to male, but it is more female in spawn periods temprorally; female and male amplectant time ranges from a few minutes to 40 minutes. After the end of the female and male amplectant, they start to spermatophore transportation. Egg-laying behavior is carried out in 1﹣2 d after, and the eggs are produced on the bottom of the breeding ground, grass or stone. The hatching rate of eggs is 55%, the average hatching time is 8 d, and the larvae can complete metamorphosis at 130 d. These studies have shown that, the reproduction and development of T. kweichowensis are greatly affected by changes in rainfall, water volume, temperature, and the breeding habitat are relatively fixed and more susceptible to human activities. Therefore, we should pay attention to the habitats protection of T. kweichowensis in the process of rocky desertification control and ecological restoration. If necessary, we should build a stable breeding habitats manually to ensure its ecological reproduction.

Abstract:After 53 (28 ♂♂, 25 ♀♀) adults of the Ornamented Pygmy Frog (Microhyla fissipes) had been collected from Mengla County of Xishuangbanna Dai Autonomous Prefecture in Yunnan Province, China, 16 morphological characteristics of them, such as body length, head length, head width, body mass and female’s brood amount had been measured. With the independent sample t test and covariance method, sexual differences of the all morphological characteristics of this frog have been tested, and the correlation between local morphological characteristics with body length of male and female adults, also the brood amount with the local morphological characteristics of female adults have been also analyzed by the linear regression analysis method. The results showed that, the average body length of female M. fissipes was 25.08 ± 0.40 mm, and the male’s was 24.78 ± 0.31 mm, there was no significant difference in body length and body mass between males and females (P > 0.05, Table 1), the size of male and female individuals was basically identical. All local morphological features were extremely significant positively correlated with body length (P < 0.01, Table 1). Length of lower arm and hand, and hand length were significantly different between males and females (P < 0.05, Table 1), there were also significant differences with the growth rate of body length. Separately, the brood amount was positively correlated with body length, body mass, inter-orbital space, diameter of lower arm, tibia-width, length of foot and tarsus (P < 0.05, Table 2), and extremely significant positive correlation with body mass (P < 0.01, Table 2). Our analyses suggest that, the sexual dimorphism of M. fissipes adults between female and male were mainly manifested in the head width, forearm length and hand length, which possibly related to the ability to obtain food for survivaling, and the success rate of males in competing for mating rights. However, individuals of female may increase their reproductive output by increasing their body length and body weight correspondingly increasing abdominal volume, which was consistented with the fecundity selection hypothesis.

Abstract:The illegal trade in wild animals has caused great damage to wildlife resources in China. By analyzing the temporal and spatial characteristics of 179 raptor poaching cases from 2011 to 2018 in east China (37.8% of the total raptor poaching cases in the same period of China), we could understand the development trend of the cases. In this study, we obtained the data set of species, the number of individuals, time and location through the statistics of judicial appraisal cases in the archives of State Forestry Administration Forest Public Security Judicial Identification Center. Excel 2016 and ArcGIS10.2 were used to carry out descriptive analysis of spatiotemporal characteristics of the cases. 35 species were involved, with a total of 530 individuals, accounting for 40.5% of the total raptor poaching individuals in China (Table 1). Nine species were the most poached (individual and case number ≥ 9), all of which were national Ⅱ key protected wild animals. In the past 8 years, the number of cases, species and individuals showed an increasing trend year by year. Spring, autumn and winter were the periods of high incidence of cases (Fig. 1). Jiangsu (23 species, 69 cases, 50 sites) and Zhejiang province (21 species, 66 cases, 31 sites) had more cases than Anhui (15 species, 23 cases, 16 sites) and Shandong province (13 species, 21 cases, 7 sites) (Fig. 2). The cases were concentrated at the provincial boundaries (82 cases), particularly close to main roads (83 cases) and railways (70 cases). We suggest that the border of two provinces should be taken as the key monitoring areas of raptor protection, by strengthening the supervision of road and water transportation, especially in spring, autumn and winter.

Abstract:Cervus nippon hortulorum was once considered to be extinct in the wild. In recent years, some small size populations were found in the southeastern part of Heilongjiang province and the eastern part of Jilin province, nearing the border. Since the habitats of C. n. hortulorum are narrow, isolated and fragmented, it is an urgent need to further evaluate the genetic changes of the population, especially the genetic diversity and inbreeding decline, so as to enhance the pertinence of conservation and management. In this study, 673 suspected fecal samples of sika deer were collected from 9 key research areas in Daxing'an, Xiaoxing'an and Changbai Mountains (Fig. 1). Firstly, species identification was carried out based on DNA Cyt b gene sequencing technology, and the positive samples were supplied for further individual identification by microsatellite technology. At last, Microchecker 2.2.3 software was used to detect the invalid allele or allele deletion of each locus; and Genalex 6.0 software was used to calculate the population average allele number (Na), observed heterozygosity (Ho), expected heterozygosity (He) and fixed coefficient (Fis). A total of 33 individuals (20 in Muling nature reserve and 13 in Laoyeling nature reserve) were identified from 106 fecal DNA samples of sika deer (Fig. 2 and Table 1). Six variation sites and five haplotypes were detected in the Cyt b sequence of these 33 individuals. The values of Hd, Pi, Na, Ho, He and Fis were 0.621, 0.006 7, 7.1, 0.604, 0.712, and 0.152, respectively (Table 1). The results showed that the population genetic diversity of Northeastern sika deer was rich, but there was also a certain degree of heterozygosity deficiency and inbreeding (Table 1); the population experienced bottleneck effect in recent years, without population expansion (Table 1 and Fig. 5); there was no genetic differentiation between populations, which could be protected as a management unit (Table 2 and Fig. 3, 4). It is suggested that the individuals with rare haplotypes should be taken as the key point in monitoring and protection, and the artificial bred populations should be released in the field at the right time, so as to improve the gene exchange between individuals in the field and accelerate the population restoration.

Abstract:Chiroptera has been identified as a natural reservoir of emerging infectious diseases for carrying zoonotic viruses. An astonishing amount of genetic diversity of viruses has been isolated from the bats in different populations throughout the world. The diversity resource of chiropteran species in Guizhou province is abundant, consisting 65 species, 19 genera and 7 families of 2 suborders. However, there has been limited research on their zoonotic viruses. In this study, 253 faecal samples and 258 bat tissue samples were collected in Guizhou Province, including 10 species in 2 families, 6 genera (S1). Based on viral metagenomics and sRNA virus detection, this research identified the viruses carried by four species of bats that are widely found in Guizhou Province. By analysing different types of bat viruses, and comparing the species differentiation between bats from Guizhou and those from Yunnan and Guangxi, the study detected four types of viruses in four bat species, including vertebrate viruses, insect viruses, plant viruses and bacterial viruses. Furthermore, there were totally 53 families, 111 genera and more than 170 virus species found, including 9 families, 10 genera and 46 species with public health significance, such as Human herpesvirus 1, Hepatitis E virus, Human papillomavirus type 16 (S2). The comparison of virus species among the three regions revealed that only Adenoviridae and Retroviridae were found in the bats inhabiting in all the three provinces, which might indicate that there could be large differences in types of bat viruses distributed in different regions. Since Chiropterans in Guizhou are found to carry a wide range of virus species which have certain representativeness in public health, it is crucial to carry out further molecular epidemiological investigations.

Abstract:The purpose of the experiment was to induce the differentiation of osteoblasts and chondrocytes from bone marrow mesenchymal stem cells (BMSCs) in the horse (Equus caballus). BMSCs were obtained by primary cell culture, and stem cell characteristics of the third generation (P3) of purified cells were identified. Then, the BMSCs were induced to differentiate in vitro, and the differentiated cells were stained and identified for specific gene expression. The results showed that the obtained horse bone marrow cells expressed stem cell transcription factors and mesenchymal stem cell surface markers. The cells obtained were thought to be horse BMSCs. After induction culture, P3 generation cells changed in shape from "long spindle" to "bone nodules" osteoblasts and “paving stones” chondrocytes. Alizarin red staining showed that the induced red osteoblasts, and the red "bone nodules" gradually increased with time. The number of Alsine blue stained chondrocytes, which were rich in proteoglycan and hyaluronic, increased with extended induction time, while no staining was observed in the control group. The expressions of Col and ALPL genes in osteoblasts changed significantly with the induction time. Common PCR results showed that chondrogenic genes such as collagerⅡ, aggrecan and Sox9 were amplified in the induced chondrocytes, while the control cells did not express these specific genes. In conclusion, BMSCs were successfully induced to differentiate into osteoblasts and chondrocytes, providing experimental basis for the repair of bone tissue defects and the treatment of chondrocytes.

Abstract:Black-necked Crane (Grus nigricollis) is a national first-class protected wildlife. Due to its harsh natural environment and small population size in the wild, the Black-necked Cranes have been bred in captivity in many zoos and protected areas for protection, however, in low reproduction rate. In order to improve its reproduction rate, we explored the physiological characteristics of Black-necked Crane in captivity during breeding period using non-invasive sampling methods and behavioral observations. The enzyme-linked immunity was used to measure the contents of sex hormones (estradiol, progesterone and testosterone). SPSS was used to conduct one-way ANOVA and multiple comparisons on the data. The significance level was set as P = 0.05. The homogeneity of sample mean variance was determined by Levene’s homogeneity of variance test, Dunnett''s T3 test was used for multiple comparisons of data from different sampling periods. The results showed that: The estradiol level of Black-necked Crane in the breeding period was significantly increased, which could be used as one of the important physiological indicators to effectively monitor the physiological status of females in the breeding period (Fig. 1); The testosterone level of males in the early stage of reproduction was significantly increased, which was significantly different from that in the non-breeding period and other stages of breeding period, indicating that testosterone could effectively monitor the reproductive physiological state of males and provide a theoretical basis for the selection of appropriate time for artificial extraction (Fig. 2 and 4); The progesterone level in the middle reproductive period of the females with successful mating was significantly higher than that in other stages, indicating that this indicator can effectively monitor the mating status of the females (Fig. 1). In conclusion, monitoring the appropriate physiological parameters can provide a theoretical basis for the breeding of captive Black-necked Crane and improve its breeding rate.

Abstract:The morphological characteristics of hemocytes in peripheral blood were investigated in Theloderma corticale by the Wright-Giemsa staining method and manual counting of blood cells．The results showed that the peripheral hematocytes of the T. corticale were composed of erythrocytes, neutrophils, basophils and eosinophils, lymphocytes, monocytes, and thrombocytes (Fig. 1). Erythrocytes mostly were oval, elliptical and few were teardrop-shaped, pear-shaped, comma-shaped, spindle-shaped cells, and their mean density was 2.43 × 105 cells/mm3. Occasionally, erythrocytes undergoing direct division or mitosis were observed. The mean of leucocytes was 1.74 × 104 cells/mm3. The proportions of small lymphocytes, large lymphocyte, monocytes, neutrophils, basophils and eosinophils were 40.02% ± 1.77%, 10.83% ± 2.53%, 23.17% ± 3.16%, 10.08% ± 4.62%, 9.25% ± 2.69% and 6.72% ± 1.50%, respectively. Single thrombocyte was ovoid or spindle-like in shape and cluster thrombocyte was round and irregularly round in shape (Fig. 2 and Table 1).

Abstract:Pigment cells are the basis of skin pattern formation. In order to understand different types and arrangement of pigment cells in different skin pattern areas of mandarin fish (Siniperca chuatsi), the skin from pattern area, non-pattern area and their junction area were observed with light and electron microscopes. The results showed that the skin contained melanophores, xanthophores, erythrophores and iridophores, which were mainly distributed in the epidermis and pigment layer. The pattern areas include eye stripe, trunk band and trunk spots that contained melanophores in both the epidermis and pigment layer. The skin non-pattern area only contained a small amount of melanophores in the dermis (Fig. 2). In the skin trunk pattern area (band, strips), different pigment cells were obviously distributed in order in pigment layer: from the outside to the inside, xanthophores, erythrophores, melanophores and iridophores were distributed (Fig. 5b), and the reflective platelets in iridophores were long and neatly arranged horizontally (Fig. 5e). In the non-patterned areas of the trunk xanthophores, erythrophores and iridophores were observed from the outside to the inside (Fig. 5c), and the reflective small plates in iridophores were short and irregularly arranged (Fig. 5f). Pigment layer of the eye stripe contained these four kinds of pigment cells, and the number of pigment cells which were irregularly arranged in this region was small, and melanin granules were large in size (Fig. 5a). At the skin junction area, the number of melanophores in the pigment layer gradually decreased toward the non-patterned area, while the number of iridophores gradually increased (Fig. 5d). The result showed that the types, distributions and arrangements of pigment cells were distinctly different among pattern areas, non-pattern area and junction area. Results of this study provide some basic information for understanding pigmentation patterning mechanism in mandarin fish.

Abstract:The distributions of nonspecific esterase, acid phosphatase and alkaline phosphatase in digestive tract of Tilapia Galilaea (Oreochromis niloticus ♀ × O. aureus ♂) were studied by paraffin section combined with histochemical methods including Gomori’s staining and acid-α-naphthyl-acetate esterase staining. It was found that nonspecific esterase densely distributed in the foregut, massively distributed in the stomach, esophagus and midgut, and slightly distributed in the hindgut (Fig. 1). Acid phosphatase mainly distributed in the foregut, followed by the midgut and stomach, and few in the esophagus and hindgut (Fig. 2). Alkaline phosphatase was densely observed in the foregut and midgut, and a larger amount was also observed in the stomach, esophagus and hindgut (Fig. 3). The results showed that the foregut, midgut, esophagus and stomach of Tilapia Galilaea were the main sites of lipid absorption and digestion. The foregut, midgut and stomach were the main sites of protein absorption and digestion, and the foregut and midgut were the main sites of nutrient absorption.

Abstract:To comprehensively understand the nutritional value of postpartum parents of Alosa sapidissima, We investigated the basic nutritional compositions, amino acid and fatty acid by biochemical analysis methods. The experimental data were statistically compared with variance analysis. The data of amino acid and fatty acid compositions and contents in fodder were listed in Table 1. The results showed that the crude protein content was significantly lower in the female parent than in the male parent (P < 0.05, Table 2), while the crude fat content was significantly higher in the female parent than in the male parent (P < 0.05, Table 2). There was no significant difference in the contents of moisture and ash between the female and male (P > 0.05, Table 2). Eighteen kinds of amino acids were detected in postpartum parents, and the amino acid content was significantly lower in the female parent than in the male parent (P < 0.05, Table 3) except for glycine and cystine. The contents of total amino acids, essential amino acids, half-essential amino acids, non-essential amino acids and delicious amino acids were significantly lower in the female parent than in the male parent (P < 0.05, Table 3). Based on the amino acid scores, the first and second limited amino acids of female parent were valine and tryptohan, respectively, but the first and second limited amino acids of male parent were tryptohan and valine, respectively (Table 4). Based on the chemical scores, the first limited amino acid of postpartum parents was tryptophan, and the second limited amino acids were methionine and cystine (Table 4). The essential amino acid indexes (EAAI) were 81.60 and 82.64, respectively in female parent and male parent (Table 4), while Their F values were 2.20 and 2.23, respectively (Table 4). Eleven kinds of fatty acids were detected in postpartum parents. There were significant difference in the contents of C18:0, C20:0, C16:1 and C18:3n between the female and male (P < 0.05, Table 5). The average contents of docosahexenoic acid (DHA), eicosapentaenoic acid (EPA) and ω3 poly unsaturated fatty acids (∑PUFA) in female parent were much lower than in male parent. In conclusion, the nutritive value of muscle in postpartum parents of A. sapidissima was at a low level, the nutritional value of male parent muscle was better than that of female parent. The data are important for improving the feeding conditions and management of A. sapidissima parents and for their intensive culture.

Abstract:The high morbidity and mortality of captive Forest Musk Deer (Moschus berezovskii) due to the digestive tract diseases, especially the intestinal inflammatory diseases have been a major problem. Fecal detection is a basis for evaluating the health status of the digestive system of the wild animals and the clinical diagnosis of the intestinal inflammatory diseases. The fecal occult blood has a wide range of clinical diagnostic values in the diagnosis of gastrointestinal bleeding. In this study, diluted fresh Forest Musk Deer blood was utilized to explore the sensitivity ranges of three methods (pyramidon chemical method, o-toluidine chemical method and benzidine chemical method) for the determination of the blood concentration in the Forest Musk Deer feces. The results showed that the minimum sensitivity test concentration of the pyramidon method was 0.05 mg/L, more sensitive than that of o-toluidine method (0.4 mg/L) or benzidine method (100 mg/L). The positive rate of gastrointestinal bleeding revealed by the pyramidon method was significantly higher when compared to the other two methods, with the positive rate of pyramidon method 10.13%, o-toluidine chemical method of 2.56% and benzidine chemical method of 0 (P < 0.05). The operation of the pyramidon method was more convenient and rapid. Therefore, the detection of fecal occult blood in the Forest Musk Deer by pyramidon method is more accurate and convenient.

Abstract:Conspecific brood parasitism (CBP) is widespread in Anseriformes. For the Scaly-sided Merganser (Mergus squamatus), however, there is no report on the occurrence of CBP. On 13th of April of 2019, in our study area at the Manjiang, Fusong in the Changbai Mountains of China, we found one female of Scaly-sided Merganser breeding in the nest-box with 15 eggs in the nest (Fig. 1a, b). During April to May, we monitored the nest by video camera, and found that one parasitic female intruded into the nest when the parent female was incubating, and stayed inside the nest box for 29 min (Fig. 1c, d). The host female showed aggressive behavior against the parasitic female (Fig. 1e), but no egg rejection occurred after being parasitized. Thereafter, an increase in clutch size from 15 to 18 eggs was recorded (Fig.1b, f). After 11 ducklings leaving the nest-box, 7 eggs were left unhatched (Fig.1 g, h) and were immediately collected and checked, among which 5 were infertile and 2 were artificially hatched and released to the wild (Fig. 1i). Based on combined evidence from an increase in clutch size, the abnormally large clutch, and asynchronous hatchling, we suggest the occurrence of CBP in this nest. At our study area, there were still 22 nest-boxes unoccupied, the causes of CBP in Scaly-sided Merganser need further investigation.

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