Abstract:[Objectives] Bamboo forest thinning is one of the effective ways of bamboo forest regeneration. To determine the effects of thinning on the regeneration of Bashania bamboo forests and foraging selection of Giant Pandas (Ailuropoda melanoleuca), we conducted a three-year (2017-2019) monitoring on the belt thinning plots with a size of 30 m × 60 m (Fig. 1) each in the Bashania bamboo forest, which is located in the non-major foraging areas (with old and dense bamboo stems) of Giant Panda in the Foping National Nature Reserve of Shaanxi Province. [Methods] We obtained in total 10 102 group data of bamboo regeneration, 1 119 Giant Panda feeding bamboo data and 32 215 infrared camera trapping data. By using SPSS t test, we analyzed the difference on bamboo regeneration between the belt thinning plots and the control plots with P < 0.05 representing a significant difference and P ≥ 0.05 representing no significant difference. We also counted the numbers of selected bamboo shoots, mature bamboos and produced feces by Giant Pandas, and calculated the occurring frequency of Giant Pandas and its sympatric animals based on infrared camera photos in order to insight the impacts of bamboo thinning on its regeneration and Giant Pandas’ food selecting. [Results] Our results showed that (1) about bamboo shoots, theie number increased significantly in the thinning plots, and the basal diameters (Fig. 2) and heights (Fig. 3) in the belt thinning plots are significantly smaller than the ones in the control plots (Except height of bamboo shoots in the 1st year), and no significant difference in the 3rd year. (2) About the mature bamboo stems, their number increased also significantly in the thinning plots, and the basal diameters (Fig. 4) and heights (Fig. 5) in the strip thinning plots are significantly smaller than the ones in the control plots (Except height of mature bamboo in the 2nd year), and no significant difference in the 3rd year. (3) The Giant Panda showed a preference to the thinning plots with more feeding on bamboo shoots and young bamboos. The numbers of bamboo stems selected and feces deposited in the thinning plots were significantly higher than those in the control ones (Table 1). Giant Pandas show a longer duration of staying in thinning plots (Fig. 6). (4) The total 23 species including Giant Panda and other sympatric species through camera trapping in the thinning plots (Fig. 7), and the highest occurrence of the Giant Pandas in the months of January, April, May and December with feeding on bamboo shorts and leaves. [Conclusion] Therefore, 2 m-width strip thinning can change the age structure of bamboo forest, improve its ageing problem and increase the range of suitable habitat for giant panda forging. We can gradually generalize this strip thinning technique in the habitat of Giant Pandas with seriously aging bamboo forest, and can be applied in protection and management of the food base areas of the captured Giant Pandas.

Abstract:[Objectives] The Leopard Cat (Prionailurus bengalensis) and Yellow-throated Marten (Martes flavigula) are both the national second-class protected species. However, as sypatric species, their ecological niche and coexistence mechanism are nevertheless unclear. [Methods] From September 2013 to December 2018, 37 infrared camera traps were set up in the Foping National Nature Reserve in Shaanxi Province. Our study area and camera trap sites can be seen from Fig. 1. Based on the monitoring data, we calculated the relative abundance index to model the cluster pattern, daytime-and-night activity rhythm, seasonal dynamics and spatial distribution of these two small and medium sized carnivores. Seasonal activity rhythm were tested by t-test and spatial activity pattern were tested by Wilcoxon rank sum test. Daily activity rhythm was modeled by kernel density method and inverse distance weighted method (weight = 5) was used to draw spatial activity pattern. All the analysis were conducted in R 3.6.3. [Results] There were a total of 72 076 camera working days and captured 290 independent and effective incidents of Leopard Cats and 84 independent and effective incidents of Yellow-throated Martens. The results showed that the Leopard Cat is a typical solitary species, preferring activity in the night (daytime-and-night relative abundance index, DRAI = 77.4%). While the Yellow-throated Marten is more gregarious and more activity in the day time (DRAI = 94.1%) (Fig. 2a). Leopard Cats reach their activity peak in a day between 3:00 and 4:00 am and the activity peak of Yellow-throated Martens appeared at 17:00 (Fig. 2a). The relative seasonal abundance index SRAI of Leopard Cats is significantly higher in the cold season than in the warm season (t = 2.82, df = 11, P < 0.05), while the SRAI of Yellow-throated Marten is significantly higher in the warm season than in the cold season (t =﹣3.09, df = 11, P < 0.05) (Fig. 3a). The monthly relative abundance index MRAI of the two species showed obvious seasonal fluctuations (Fig. 3b). There was significant mutual avoidance between Leopard Cats and Yellow-throated Martens in spatial distribution. However, Leopard Cats is more frequently recorded in the area active moved more frequently in areas closer to human paths than Yellow-throated Martens (Fig. 4a). The camera site abundance of index CRAI to the road within 500 m is significantly higher for Leopard cat’s than that of Yellow-throated Marten (Distance, Wilcoxon rank sum test: P < 0.001) (Fig. 4b). [Conclusion] The two small carnivorous mammals occupied different spatial and temporal ecological niches to reduce competition and achieve coexistence in a large region.

Abstract:Damage or loss from bird activity is a worldwide concern, impacting many industries (including agriculture, forestry, fisheries, and electric power distribution), aircraft safety, epidemic disease transmission, and human health. Bird damage can cause huge economic losses, and it is difficult to prevent and control. [Objectives] Our goal is to compile a list of birds which damages to crops in Xinjiang Provence, the northwest of China. On the basis of understanding their patterns of activity, we propose effective control measures. [Methods] Our study was based on transect censuses, questionnaires, bird counts in mist-nets, and bird stomach contents. We set up observation points in Moyu, Wushi, Baicheng and Changji counties, with a latitudinal span from N 37° to N 44°, and a range of habitat types including farmland, orchard and reservoirs, over the period from 2019 to 2021. Different hazard levels were preliminarily divided through field observations, bird quantity census, feed intake and questionnaire analysis (n = 881 persons or families), (Table 1). [Results] A total of 49 bird species were recorded in 40 genera, 19 families and 10 orders harmful to agriculture, which has exceeded more than 10% of the number of bird species in Xinjiang. These species mainly damaged corn, wheat, rice, grapes, dates, oil sunflower, potatoes, watermelons, common melons, walnuts, strawberries, and Chinese wolfberry. Among these losses, winter wheat (52.9%) and wine grape (14.4%) were the most significant. For the whole investigation area, the probability of bird presence is about 28.6% (Table 2), which decreases from north to south (R2 = 0.893, P < 0.05). In some places, it is estimated that the harvest could be damaged by approximately 26.6%. Only 18.0% of farmers use big horns, mist-nets, scarecrows, chemicals, or other measures to take preventive measures. At present, insurance claims and government subsidies are very small, less than 9.53% (sometimes no any relief or compensation). In order to prevent harm, some farmers have set up sticky bird nets or traps in the farmland. The most common birds caught by the nets were starlings (44.8%) and sparrows (28.4%). The diet of starling which the so-called beneficial bird in China has changed greatly, which is closely related to excessive reclamation and heavy use of pesticides. In addition, sparrows and starlings not only made harms to cereal crops, but also to fruit crops. The conflict between birds and humans has lasted for thousands of years, and in 1950s, there was even a great national movement to eliminate “sparrows” in China. [Conclusion] Bird damage on crops and fruits is related to food security, so we must pay attention to it. Finally, we discussed the measures to repel birds, including biological control, chemical expulsion, and physical intimidation (See pictures). In fact, to prevent bird damage effectively and harmlessly during the bird repellent is also a way to protect the birds.

Abstract:[Objectives] Chinese Alligator, Alligator sinensis, listed as the first level protected animals in China, is an ancient and rare reptile which is endemic to China. There is no report on the cooling treatment for hibernation in North China. In this study, the 5 Chinese Alligators reared in Beijing Zoo were artificially cooled for hibernation during November 2020 to May 2021, in order to understand the characteristics and environmental needs of hibernation and the relationship between the environmental temperature and the behavior, also to explore the behavior ethogram during hibernation and establish a suitable way of controlling temperature for hibernation, which will provide a reference for safe overwintering and feeding management in North China. [Methods] The simple planar graph of the environment shows in Fig. 1. During the study, the environmental selection and behaviors of the Chinese Alligators were observed in each hibernation periods, the body temperatures were monitored during deep hibernation period, and the changes of behavioral indicators and biological indexes before and after hibernation were analyzed. ANOVA was used to analyze the variance between different distribution areas. Paired-Samples T Test was conducted to analyze the difference before and after hibernation and the difference periods. [Results] The animals were in primary hibernation when the average room temperature were 13.00 ± 3.10 ℃ (range 9.4﹣20.0 ℃) (Table 1, Fig. 2). The animals were in deep hibernation when the average room temperature and water temperature were 11.35 ± 2.60 ℃ (range 7.0﹣ 13.0 ℃) and 9.60 ± 2.09 ℃ (range 6.0﹣11.0 ℃) respectively, when the average room temperature was 12.17 ± 1.95 ℃, the animals were in deep hibernation, and when 10.54 ± 3.01 ℃ the minor reaction to human and external interference. The animals were in recovery period when temperature were 15.50 ± 1.77 ℃ (range 12.0﹣18.4 ℃). The body temperature of the animals was 10.9 ± 0.55 ℃ during the deep hibernation, and the whole hibernation period was nearly 7 months. During hibernation period, the inactivity behaviors mainly includes hibernating, piling up and hiding, the activity behaviors mainly includes crawling, diving, swing tail, open eyes, shaking head, rising head in water, putting head on something, warning exhalation and roaring. The percents of the inactivity behaviors frequency were 46.5% in the primary period, 76.0% in deep period, and less than 10% in the recovery period (Table 2). Most of the behaviors have significant differences between each hibernation periods (P < 0.05). There were individual differences in habitat selection in each hibernation stage. In the primary hibernation period, most individuals were on the land (89.4%). In the deep hibernation period, most individuals mainly hibernated in water and occasionally on land (75.8% in water), while all individuals were prefer to the pool during recovery period (90.4% in water) (Fig. 3). The distribution sites of the Chinese Alligators in land-area were mainly in the depressions area around the plants, which were consistent in each hibernation period (Fig. 4). In the water, they were mainly in the area where was less interference and with the low stones on the edge of the pool, the distribute percent was more than 95% (Fig. 5). There was no significant difference in of the biological indexes before and after hibernation (P > 0.05, Table 3). After hibernation, they began to eat at 26.0﹣28.0 ℃. [Conclusion] The results showed that, the Chinese Alligators were in hibernation when the average room temperature and water temperature were kept at 7.0﹣13.0 ℃ and 6.0﹣11.0 ℃ respectively in North China. By artificial cooling and enriching the habitat types of hibernation environment, the animals could be safe overwintering in hibernation, and the environmental temperature, humidity and environmental stability were the important factors to hibernation.

Abstract:[Objectives] Breeding period occupies an important stage in the life history of cephalopods, we study their reproductive needs by observing their behaviors. [Methods] In June 2020, we observed directly on 30 Octopus sinensis reared in an indoor pool during their breeding period as well as by underwater photography. [Results] The behaviors such as movement, predation (Fig. 1), courtship, mating (Fig. 2), oviposition and brooding (Fig. 3) were found during the spawning period. The power to conduct swimming and crawl in O. sinensis was produced by blowing water with their funnel to move short distanceforward, and they retreated quickly when they were shocked and occasionally as well as jetting ink. Before spawning, O. sinensis actively hunted and consumed Charybdis japonica. After spawning, the appetite of females decrease and finally stopped eating and males lost their appetite gradually. Courtship and mating occurred frequently during spawning period. Males pursued females actively and tended to mate with females with the similar body size, and they also to fight against each other for mating. O. sinensis was polygamy, and kept a certain distance when mating. Males and females turned their body dark brown during mating, and males extend their arms to keep other males away from their mating female. Males could mate with different females, and vice versa. There was a phenomenon of one female mating with multiple males at the same time. Males matured earlier than females, rather than laying eggs immediately after mating, females usually waited for maturation of gonads. Some individuals begun spawning even after more than 50 d. O. sinensis tended to lay their eggs in their nests and sealed them by stones. If there was no nest, the fertilized eggs were protected by covering them with their bodies. Females cleaned the dead eggs and blew the fertilized eggs to provide oxygen after spawning. The fertilized eggs were translucent yellow and elastic, the average diameter was (2.5 ± 0.2) mm × (1.1 ± 0.1) mm. The average fecundity was 83 988 eggs (Table 1). Females had the brooding behavior, the incubation time was between 21 and 24 days under 22.4﹣23.5 ℃. After hatching, the body weight of female was 46% lower than that before oviposition, and died gradually. The longest survival time of brooding female was 34 d, and the longest survival time of mating male was 56 d. [Conclusion] We conclude that the polygamy of O. sinensis could improve their mating success rate, and the brooding behavior could benefit to larval hatching.

Abstract:[Objectives] In order to reveal the differences in the main morphological characteristics of the Leptobrachella, it is hoped to provide a reference for the subsequent taxonomy and pedigree geography of Leptobrachella. [Methods] Apply morphological analysis methods, use the shortest distance method of Euclidean distance for cluster analysis, construct a cluster tree to show the grouping relationship of six species. The stepwise discriminant method is used for discriminant analysis, the trait parameters that contribute greatly to the discrimination are selected, and the discriminant formulas of six species of Leptobrachella are established. The morphological characteristic values of the individuals to be determined are respectively substituted into the discriminant formulas, and the formula values are calculated and compared. The largest is the population to which the individual of Leptobrachella belongs, so as to verify the ability of each main deformation contribution trait to distinguish species. [Results] The results indicated that: Morphological character Snout-vent length comparison: From large to small, the snout-vent length of the six species of Leptobrachella are L. chishuiensis, L. suiyangensis, L. jinshaensis, L. bijie, L. wulingensis and L. purpuraventra (Table 3); Cluster analysis showed that among the 6 species, the difference between the L. purpuraventra and the L. bijie was the largest, and the L. purpuraventra and the L. chishuiensis had the smallest difference (Fig. 2); The difference of six species of Leptobrachella mainly focused on the ratio of snout length / snout-vent length, internasal distance / snout-vent length, upper eyelid width / snout-vent length, maximal tibia width / snout-vent length, hindlimb length / snout-vent length, fourth finger length / snout-vent length (Fig. 4). The accuracy rate of morphological feature discrimination was 75.0%﹣100.0%, and the comprehensive discrimination accuracy rate was 90.0%. In addition, we also compared the quality characteristics of six species of Leptobrachella (Table 4), and found that they can also be distinguished by quality characteristics. [Conclusion] This study initially revealed the morphological differences of these six species of Leptobrachella in the Guizhou Plateau. It is believed that the Leptobrachella in the Guizhou Plateau can be well distinguished by measurable and qualitative traits.

Abstract:[Objectives] The Wuyi Mountain and Meihua Mountain National Nature Reserves in Fujian Province are located in the mid-subtropical monsoon evergreen broad-leaved forest climate zone. This region has high species diversity. However, there are relatively few reports on the species diversity of soil animals (earthworms). We thus conducted an earthworm collection in the Wuyi Mountain and Meihua Mountain National Nature Reserves. [Methods] Morphological taxonomy methods and DNA barcode technology were used to identify the new species. The Bootstrap and P-distance methods were used to calculate the genetic distance between species, and the Bootstrap method and Jukes-Cantor model were used to construct molecular phylogenetic NJ trees in MEGA-X. [Results] Four new earthworm species belonging to the family Megascolecidae and genus Amynthas were found: A. epigenus Sun & Qiu, sp. nov., A. meihuamontis Sun & Qiu, sp. nov., A. muscosus Sun & Jiang, sp. nov. and A. humatalis Sun & Jiang, sp. nov (Fig. 1﹣4). These four new species belong to the A. sieboldi-group in the species group of Sims & Easton, and their spermatheca pores are located in 6/7﹣8/9. We provided morphological descriptions, pictures, morphological comparisons with similar species, and barcode accession numbers submitted to GenBank for all the new species. In addition, we also searched out earthworm species that are closely related to the new species through BLAST on GenBank (Table 1). The genetic distance calculations show that the genetic distance between new species and related species is greater than the interspecies genetic distance (usually less than 1%), and meets the range of intraspecies genetic distance (8.3%﹣13.0%, and above), so they have at least species level Genetic difference (Table 2, 3). The molecular phylogenetic relationship rebuilding result shows that the positions of the four new species in the phylogenetic tree are different from those of other species, which further proves that the four species reported in this paper are new species (Fig. 5). [Conclusion] In summary, the morphological and genetic evidence suggests that A. epigenus Sun & Qiu, sp. nov., A. meihuamontis Sun & Qiu, sp. nov., A. muscosus Sun & Jiang, sp. nov. and A. humatalis Sun & Jiang, sp. nov are new species. The present report has made the number of earthworm species increase from 4 to 8 in Wuyishan and Meihuashan National Nature Reserves in Fujian Province.

Abstract:[Objectives] The radula of the terrestrial mollusk is generally composed of a central vertical tooth row, several vertical rows of lateral teeth on both sides, and several vertical rows of marginal teeth on both sides of the lateral teeth. The aim of the present study was to analyze the radulae characteristics of different families, genera, species of terrestrial mollusks, and to further explore the taxonomic significance of radula form classification hierarchies of terrestrial mollusks. [Methods] With the back of the terrestrial mollusk facing upwards, the mantles on both sides were separated, the buccal mass was taken, and the muscle tissue was removed toobserve the radula sac. The radula sac was treated with 5% NaOH solution for 3 to 5 minutes, followed by peeling off the radula and removing the connective tissue, then the alkaline solution on the radula was wash off with distilled water, followed by ultrasonic cleaning. After dehydration with alcohol gradients, samples were stored in 70% alcohol. The radulae were transferred to the sample table for metal spraying after natural drying, and were observed and photographed under the FEI Quanta 200F scanning electron microscope (SEM). The radulae of 17 species of terrestrial mollusks belonging to 3 orders, 10 families, 12 genera were observed. [Results] The results showed that among the 17 species of terrestrial mollusks studied, the number of vertical rows of central teeth was all one. The numbers of vertical rows of lateral teeth ranged from 12 to 218, and the numbers of marginal teeth ranged from 0 to 204. The central teeth were divided into unicuspid, tricuspid, and multitooth types (Fig. 5). There were many types of lateral teeth and marginal teeth. The numbers of cusps of lateral teeth were from 1 to 6, and the numbers of cusps of the marginal teeth varied from 1 to 10. [Conclusion] Combined with the 38 species of terrestrial mollusks previously reported, there are 55 species in total in 3 orders, 20 families, and 41 genera, and the morphology and numbers of cusps of the central teeth, lateral teeth, and marginal teeth of terrestrial mollusks of different families are quite different. Radulae of the same genus have similarities in morphology, and the difference in morphology is small. The numbers of vertical rows of different species in the same family are quite different, and there is no regularity (Appendix 2). The morphological characteristics of radula have important taxonomic significance in different classification hierarchies.

Abstract:[Objectives] Four species of amphibian were recorded in Wanfoshan Town, Tongdao County, Hunan Province, China at 26°11′47″ N, 109°56′21″ E, altitude 924 m during a filed survey on 23 May, 2020. They are recognized as first recorded in Hunan province by morphological and molecular. [Methods] [Objectives] The genus Nidirana species was endemic group of classes in Oriental realm, widely distributed in subtropical areas of East and Southeast Asia. Among the species in genus Nidirana, Nidirana adenopleura had the widest distribution area and only distributed in Taiwan, northern of Fujian, southern of Zhejiang, central Jiangxi and other regions in China. The recent contribution to the phylogeny of genus Nidirana reconsidered the previously recorded as N. adenopleura populations in southern of China were actually the genus Nidirana new species. There are several geographical population of genus Nidirana mainly distributed in the northeast, north, central and southern regions of Guangxi, had been certified the genus Nidirana population form Dupang mountain was N. xiangica, the genus Nidirana population form Dayao mountain was N. yaoica, and the genus Nidirana population form Daming mountain was N. guangxiensis. [Methods] Seven male and three female specimens collected form Rongshui were examined and measured with digital calipers. The measurements were as follows: snout-vent length, head length, head width, snout length, internasal distance, interorbital distance, eye diameter, tympanum diameter, tympanum-eye distance, hand length, radio-ulna length, foot length, tibial length, and compared characters with the N. leishanensis specimens obtained from Guizhou. All specimens were fixed in 10% buffered formalin, transferred to 70% ethanol, and de?posited in NHMG. Four muscular samples attained from euthanasia specimens and then preserved in 95% ethanol and stored at ﹣40 ℃ were used for molecular analysis. All samples were 36 sequences from all known Nidirana species and two sequences from the out-group Babina were obtained from GenBank and incorporated into our dataset (Detail information of these materials was shown in Table 1). Genomic DNA were extracted from muscle tissue samples using DNA extraction kit. A mitochondrion genes namely partial cytochrome C oxidase I gene (COI) were am?plified, and the primers were dgLCO and dgHCO. PCR amplifications were processed with the cycling conditions that initial denaturing step at 95 ℃ for 4 min, 35 cycles of denaturing at 94 ℃ for 40 s, annealing at 52 ℃ for 40 s and extending at 72 ℃ for 60 s, and a final extending step at 72 ℃ for 10 min. PCR products were sequenced with both forward using reverse primers. The Clustal W algorithm was used in MEGA 6.0 software to compare all the COI gene sequences and calculate the genetic distance between specimens and other species by the p-distance model. The comparison sequences were calculated to get the optimal nucleotide substitution model in jmodeltest v2.1.2. Sequenced data analyzed using maximum likelihood (ML) in RaxmlGUI 1.3 to structure the phylogenetic tree. Advertisement calls of the specimens were recorded in the field at the air temperature 22 ℃ using a SONY PCM D100 digital sound recorder. Praat 6.0.27 was used to obtain the oscillogram, sonogram, and power spectrum. Raven pro 1.5 was used to quantify the acoustic properties. [Results] We obtained four mitochondrial COI gene sequences length 639 bp and the optimal nucleotide substitution model for phylogenetic analysis was GTR + G + I. The Maximum likelihood phylogenetic tree of genus Nidirana showed these samples and N. leishanensis samples in Guizhou together formed a monophyletic group (Fig. 1). Genetic distance estimation based on p-distance model showed the intraspecific genetic distance was 0.2%, and the genetic distance was 0.5% between the specimens and the N. leishanensis samples (Table 2). The following principal characters of genus Nidirana specimens: a largebody size (Snout-vent length, SVL, males: 47.5 mm < SVL < 51.2 mm; females: 49.4 mm < SVL < 59.1 mm, for more please see Table 3); the presence of lateroventral grooves both on fingers and toes, relative finger lengths: II < IV < I < III; tibiotarsal articulation reaching the level between eye and nostril when leg stretched forward; a pair of subgular internal vocal sacs at corners of throat in males; nuptial pad present on the inner side of base of fingers I and II in males in breading season; webbing formula: I 2 –213 II 2 –223 III 312 –323 IV 323 –3V. Dorsal skin of head, anterior part of body, ventral surface of head and limbs were smooth. All specimens were similar in morphology but some individuals different in color pattern (Fig. 2). The advertisement call had only one strophe with one syllable. The time of syllable duration and interval syllables duration were respectively 0.18﹣0.25 s, 11.40﹣23.08 s, and the frequency of sound wave was 350﹣4 270 Hz (Fig. 3). With this wok, the morphological characteristics and acoustic characteristics of genus Nidirana population form Rongshui were accorded with N. leishanensis. We confirmed these were N. leishanensis, and it was a new record of amphibians in Guangxi. [Conclusion] This discovery extended the distribution area of the N. leishanensis from eastern Guizhou to northern Guangxi and provided important information for the phylogeography and phylogeography of genus Nidirana. We have clarified up the current species distribution of genus Nidirana in Guangxi. The complex rivers and mountain systems in Guangxi may have played an important role in isolating the communication of genus Nidirana population, promoted the population differentiation and formed new species.

Abstract:[Objectives] Four species of amphibian were recorded in Wanfoshan Town, Tongdao County, Hunan Province, China at 26°11′47″ N, 109°56′21″ E, altitude 924 m during a filed survey on 23 May, 2020. They are recognized as first recorded in Hunan province by morphological and molecular. [Methods] Compared the specimens with other species of the genus Nidirana morphologically by comparing their measurement data and diagnosis feature (Sample information is shown in Table 1). Molecularly, based on 16S rRNA gene constructed Bayesian inference tree to analyze its phylogenetic relationship, the Kamura two- parameter model in MEGA v6.0 was used to calculate the genetic distances between species of the genus Nidirana. [Results] Through this research, we found the specimens were similar with N. leishanensis in morphological, morphometric measurements (Table 2) and feature (Fig. 1) conform to diagnosis characters. Based on 16S rRNA gene constructed Bayesian inference tree showed that the four specimens were clustered with the N. leishanensis with a high support (1.00) (Fig. 2). The genetic distance between the specimens and N. leishanensis from type locality was 0.8%, much lower than that between Nidirana species range from 1.4% to 6.4% (Table 3). [Conclusion] Based on morphological and molecular compared, the specimens were identified as N. leishanensis, which is the new record of this species in Hunan Province, China.

Abstract:[Objectives] One specimen of Tropidophorus was collected from Yamugou Scenic Area, Jiangkou County, Guizhou Province during filed work in August 2019 and one specimen of Calotes was collected from Pandu Township, Lianjiang County, Fujian Provence in October 2019. [Methods] In order to identify these species, we conducted a morphological study on these specimens. [Results] After comparing the morphology of the head, body, tail and limbs, it was found that the morphological characteristics of this batch of specimens were in line with the identification characteristics of T. sinicus and C. versicolor (Fig. 1, 2). [Conclusion] The specimen from Guizhou is Tropidophorus sinicus, which is a new record of lizard in Guizhou, also the northernmost recorded distribution of this species; the specimen from Fujian is Calotes versicolor, a new record in Fujian and the most eastward record of this species.

Abstract:Melatonin is important for seasonal breeding animals to display their breeding characteristics in an appropriate season by regulating the hypothalamic-pituitary-gonadal axis. The Giant Panda (Ailuropoda melanoleuca) is a seasonal breeding mammal, usually breeding from March to May. [Objectives] To explore the seasonal changes in melatonin and testosterone in male giant pandas, [Methods] urine samples from three adult male giant pandas housed at the Chengdu Research Base of Giant Panda Breeding were collected once a week for one year (2018.8-2019.7) under natural photoperiod conditions. Depending on the local climate, spring is from March to May, summer includes June and July, autumn is from August to November, and winter is from December to February of the following year. These samples were analyzed for urinary melatonin and testosterone levels by ELISA. The seasonal changes of melatonin and testosterone levels were analyzed by one one-way ANOVA, and the differences of inter-seasonal data were analyzed by independent sample t-test. The data of testosterone levels were adjusted for homogeneity of variance by using the log value for analysis, however, all graphs were expressed in the actual testosterone levels. [Results] The results indicated that melatonin levels showed a seasonal secretion pattern (Fig. 1 and 2) and its level in winter was extremely significantly higher than that in in autumn, spring and summer, respectively (P < 0.01), and there were no significant differences among the levels in autumn, spring or summer (P > 0.05). Testosterone levels also showed a seasonal secretion pattern (Fig. 3 and 4) and its level in winter was extremely significantly higher than that in autumn or summer (P < 0.01). Testosterone level in spring was also extremely significantly higher than that in autumn or summer (P < 0.01), however no significant difference was detected between winter and spring (P > 0.05). Additionally, there was no significant difference between autumn and summer either (P > 0.05). [Conclusion] Urinary melatonin and testosterone levels show seasonal changes. Melatonin and testosterone levels increase synchronously from autumn to winter; melatonin level decreases sharply, while testosterone remains at a high level from winter to spring. The results provide reference data for further investigation of the effect of melatonin on the seasonal breeding of giant pandas.

Abstract:[Objectives] In order to explore the organizational structure characteristics of reproductive organs in mammals adapting to the plateau environment, we compared the structures of male reproductive organs between Tibetan Sheep (Ovis aries) and Small-tail Han Sheep. [Methods] The testis, epididymis, and vas deferens of adult Tibetan Sheep and Small-tail Han Sheep were collected respectively. The differences in structures of reproductive organs were compared by gross anatomy, paraffin section, and conventional H.E staining. All the data were analyzed by T-test. [Results] The results indicated that the cilia in the lumen of caput epididymis and corpus epididymis were longer, on the contrary, the cilia in the lumen of cauda epididymis were shorter and displayed a clear brush border structure in Tibetan sheep (Fig. 2). There were more smooth muscle cells in the vas deferens, the lamina propria, and the mucosal layer closely adhered, and more obvious irregular folds in Tibetan Sheep (Fig. 3). The cross-sectional diameter, area, and epithelium thickness of seminiferous tubules in Tibetan Sheep were significantly lower than in Small-tail Han Sheep (P < 0.05) (Table 1); meanwhile the diameter and area of spermatogonia and primary spermatocytes, and the number of Sertoli cells were also significantly reduced in Tibetan Sheep than in Small-tail Han Sheep (P < 0.05) (Table 1). The inner diameter, outer diameter, and cilia length of the caput epididymis, corpus epididymis, cauda epididymis were significantly reduced in Tibetan Sheep (P < 0.05) (Table 2); the thickness of the columnar epithelium of the epididymis was significantly increased (P < 0.05) (Table 2), whereas the diameter of the vas deferens lumen and the thickness of smooth muscle were significantly decreased (P < 0.05) (Table 3). [Conclusion] The study suggests that the structures of reproductive organs have undergone certain adaptive changes during the long-term adaptation process of Tibetan sheep to high altitude and low oxygen environment, which may be related to the maintenance of normal reproductive performance in the plateau environment.

Abstract:[Objectives] Animals which inhabit in plateau are facing with the unique environmental challenges such as low temperature and hypoxia. As the rising of altitude, these challenges may increase. The metabolism of animals plays an important role during long-term physiological adaptation. With the changes of altitude, the metabolic rates of many animals vary synchronously to adapt to different habitats. However, it is still unclear whether the animal thermal neutral zone (TNZ) changes with altitude. In this study, we took Plateau Pikas (Ochotona curzoniae) as studying objects to test how their thermal neutral zone range changes at different altitudes. [Methods] We examined the physiological traits of Plateau Pikas by measuring body temperature (Tb), metabolic rate and thermal conductance (C). Resting metabolic rates (RMR) were measured using 8-channel FMS portable respiratory metabolic system (Sable Systems International, Henderson, NV, USA). Biochemical incubators (LHR-250, Shanghai Yiheng Scientific Instruments Co., LTD.) was used to adjust the chambers’ temperature (Ta) within ± 0.5 ℃. Experiments were conducted under 15.0 ℃, 20.0 ℃, 22.5 ℃, 24.0 ℃, 25.0 ℃, 26.0 ℃, 27.5 ℃, 29.0 ℃, 30.0 ℃, 31.5 ℃, 32.5 ℃ and 35.0 ℃, respectively. The differences of metabolic rate among each group at ambient temperature were analyzed by repeated measure ANOVA, the differences between resting metabolic rate and body weight at different altitudes were analyzed by one-way ANOVA. [Results] The results showed that the body mass of Plateau Pikas at high or medium altitude region was significantly higher than that of low altitude region (Fig. 3). The body temperature of Plateau Pikas at three altitude regions continuously increased with the increase of ambient temperature (Fig. 1). Compared with high and medium altitude regions, Plateau Pikas at low altitude region had a lower metabolic rate (Fig. 3). The thermal neutral zone of Plateau Pikas was 24﹣30 ℃ at both high and middle altitude regions, while it was 25﹣30 ℃ at low altitude region (Fig. 2). The thermal conductance of Plateau Pikas at three altitude regions continuously increased with the increase of ambient temperature (Fig. 4). The thermal conductance was the lowest and stable when the ambient temperature was between 15 ℃ and 29 ℃, the mean thermal conductance values at high, middle and low altitude were (0.18 ± 0.09) ml/ (g·h·℃), (0.19 ± 0.09) ml/ (g·h·℃) and (0.19 ± 0.08) ml/ (g·h·℃), respectively. [Conclusion] This study found that as the rising of altitude, both metabolic rate and thermal neutral zone of Plateau Pikas increased to adapt to colder environment at higher altitude regions. These results provide basic information for understanding the physiological adaptation of animals at different altitude regions.

Abstract:[Objectives] The purpose of this study was to isolate strains capable of efficiently decomposing cellulose from the intestines of Muskrat (Ondatra zibethicus), so as to provide strain resources for the development of microbial preparations of cellulose decomposing bacteria. [Methods] Six cellulose decomposing bacteria were isolated from the cecum of Muskrat using the culture medium with sodium carboxymethyl cellulose (CMC-Na) as the only carbon source. The Congo red staining experiment and the determination of carboxymethyl cellulase (CMCase) activity were carried out. Bacterium strain WJ-3 with highly efficient cellulose decomposing activity was screened, and further identified by morphological observation, physiological and biochemical analysis as well as 16S rDNA gene sequencing approach. The 16S rDNA gene sequence was analyzed by BLAST in national center for biotechnology information (NCBI), and the phylogenetic tree was constructed by MEGA 6.0 software. The enzymatic characteristics of CMCase produced were tested to analyze the optimal reaction pH and temperature of the cellulase, as well as the tolerance of the cellulase to different temperatures and pH values. Ultimately, all the data were processed with Spss 23.0 software. [Results] A strain WJ-3 with efficient cellulose decomposing activity was identified and it was agram-positive, spore-forming, and rod-shaped bacterium (Fig. 2, Fig. 3). The physiological and biochemical test results of strain WJ-3 were shown in Table 2. By the 16S rDNA gene sequence analysis, strain WJ-3 was identified as Bacillus aerius, and named Bacillus aerius WJ-3 (GenBank Accession: MZ 292092.1). The phylogenetic tree of strain WJ-3 was shown in Figure. 5. When the CMCase produced by strain WJ-3 reacted in the range of pH 4.0﹣6.0, the enzyme activity increased with the increase of pH value, and the best reaction pH was 6.0 (Fig. 6), and the cellulase could maintain more than 80% of the relative enzyme activity after being stored in the range of pH 4.0﹣8.0 for 30 min (Fig. 7); When the CMCase produced by strain WJ-3 reacted in the temperature range of 30﹣50 ℃, the enzyme activity gradually increased with the increase of temperature, the enzyme activity was the highest at 50 ℃ (Fig. 8), and then the enzyme activity of CMCase decreased with the increase of temperature, the cellulase could maintain high enzyme activity after being stored in the temperature range of 30﹣50 ℃ for 30 min (Fig. 9). [Conclusion] The enzyme activity of CMCase produced by strain Bacillus aerius WJ-3 is relatively high, with satisfactory acid and alkali resistance as well as thermal stability. It is a strain resource with certain utilization value.

Abstract:Forest Musk Deer (Moschus berezovskii) is a kind of national first-class protected wild animal and is famous for secreting musk. The musk is a special substance secreted by the musk gland of male Forest Musk Deer. Due to excessive hunting and habitat fragmentation, the number of wild forest musk deer has dropped sharply to an endangered status. Since the 1950s, artificial breeding of Forest Musk Deer has been conducted in China, accumulated rich experienc. However, little progress has been made in both basic theories and key technologies in the many aspects including nutritional needs, feed processing and feeding management, genetic characteristics and breeding, reproduction, as well as musk secretion and extraction, which affect its population growth and musk yield. Understanding the secretion mechanism is essential to increase the yield of musk. This paper reviews the research progress on the microstructure and ultrastructure of Forest Musk Deer gland, the musk secretion process, the regulation of musk secretion as revealed by molecular markers, the relationship between sex hormones and musk secretion, and musk secretion related genome and transcriptome changes, in order to provide a reference for the study of the molecular mechanisms of musk secretion.

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Abstract:On August 11,2019, a Wilson's Storm Petrel (Oceanites oceanicus) was found in West Lake Hangzhou, Zhejiang Province. This bird is a new record for Zhejiang Provice.

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