Abstract:[Objectives] Paracanthocobitis belongs to Nemacheilidae (Cypriniformes). It is widely distributed from the Indus River basin in Pakistan to the Mekong River basin in Cambodia and Laos. Only one species P. botia,was recorded in China. During July 2022, some loach specimens were collected from Yingjiang County, Dehong Dai and Jingpo Autonomous Prefecture, Yunnan Province. Based on morphological and molecular evidence, they were conspecific with P. mandalayensis, which is a new species record to China. [Methods] In this study, the collected specimens were identified and morphological characteristics were described with reference to research literatures of Paracanthocobitis. Metric traits and vertebrae numbers were measured and counted by electronic vernier calipers and X-ray photography instrument, respectively. Based on COI gene fragments, bayesian inference tree was constructed by Phylosuite 1.2.2 and the genetic distance between individuals of Paracanthocobitis was estimated by p-distance in MEGA X. All new sequences in this study have been uploaded to NCBI GenBank, and other sequences were obtained from NCBI GenBank database (Table 1). [Results] In this paper, our material of P. mandalayensis is fully described, morphological photos (Fig. 1), morphological (Table 2) and molecular (Appendix 1) data for identification are also provided. The main features of P. mandalayensis are papillated pad with median notch on each side of lower lip; a conspicuous black spot (ocellus) close to upper extremity of caudal-fin base; presence of suborbital groove in males; complete lateral line; presence of axillary pelvic lobe; 10? branched dorsal-fin rays; 4﹣8 dark bands on caudal fin. The maximum genetic distance between the collected specimens and individuals of P. mandalayensis in Myanmar is 0.012, while the genetic distances with other species of Paracanthocobitis are 0.129﹣0.189. Therefore, based on morphological and molecular evidences, the specimens in this study are identified as P. mandalayensis. In addition, the minimum genetic distance between the collected specimens and individuals of P. mandalayensis in Thailand is 0.118. There may be cryptic species in P. mandalayensis based on phylogenetic tree (Fig. 2) and disjunction distribution, which also raised questions about the occurrence of P. botia in China. [Conclusion] This study provides a scientific basis for the investigation and conservation of fish resources in Gaoligong Mountain area, as well as new data for the identification of species of genus Paracanthocobitis in Southeast Asia.

Abstract:[Objectives] The nest predation of birds is often affected by nest-site selection. Studying the relationship between nest site and nest predation rate may help to reveal the role of different nest site characteristics in the successful reproduction of birds. [Methods] Using Black-throated Tit (Aegithalos concinnus) nests that were predated and those succeeded in the Dongzhai National Nature Reserve of Henan province from 2014 to 2017 as our study subjects, we investigated the relationship of daily nest survival rate with the found date of nest, and nesting plants by using nest survival analysis with RMark (a R package that provides a formula-based interface for the program MARK). We also assessed their nest-site safety class based on nest-site characteristics and examined whether researchers’ assessment of nest-site safety class could predict daily nest survival rate. Because nest predation might differ between different breeding stages, the egg stage (egg-laying and incubation stage; 124 nests) and nestling stage (119 nests) were analyzed separately. [Results] Daily nest survival rate decreased significantly with increasing nest age at the egg stage (Fig. 1, Table 2), but no significant relationship between nest age and nest survival rate existed in nestling stage (Table 3). In addition, daily nest survival rate did not change with the found dates of the nests and nest safety levels (Table 2, Table 3). However, at the egg stage, the nests built on bamboos (Indocalamus tessellatus, Phyllostachys sp.), Chinese firs (Cunninghamia lanceolate), bushes and grasses (e.g. Rosa spp.) and teas (Camellia sinensis) had a significantly higher daily nest survival rate than those built on cypress (Platycladus orientalis, Juniperus chinensis) and pine trees (Pinus massoniana) (Fig. 2a, Table 2). Furthermore, at the nestling stage, the nests built on bushes and grasses had a significantly lower daily nest survival rate than those built on teas, Chinese firs and bamboos (Fig. 2b, Table 3). [Conclusion] Our results suggest that nesting on certain plants may reduce the nest predation risk of the Black-throated Tit, highlighting the role of nesting plants in influencing the breeding success of birds.

Abstract:[Objectives] Cabot’s Tragopan (Tragopan caboti) is an endemic pheasant in China, and a class Ⅰ nationally key protected species. The high rate of nest egg loss has been restricting the population of wild T. caboti, and the main cause of nest egg loss is the damage related to natural enemies. [Methods] From 2010 to 2021, video monitoring technology, infrared camera monitoring and manual observation were used to investigate the natural enemies of nest eggs of 42 artificial nests used by wild T. caboti (36 nests were monitored by video, 2 nests were monitored by infrared camera, and 4 nests were observed manually) in Wuyanling National Nature Reserve, Zhejiang Province. [Results] Based on the monitoring data, we found that the clutch size of wild T. caboti was 2﹣5, the breeding success rate was 40.5% (17 out of 42 nests had at least one egg hatched), 54.8% of the nests (23 out of 42 nests) were destroyed by natural enemies, and 91.3% of the destroyed nests (21 out of 23 nests) were predated by King Ratsnake (Elaphe carinata), which is the main natural enemy of T. caboti (Fig. 1). Predators of nest eggs also include Eurasian Jay (Garrulus glandarius) and Yellow-throated Marten (Martes flavigula). Possible natural enemies include Grey Treepie (Dendrocitta formosae), Yellow-bellied Weasel (Mustela kathiah) and Stump-tailed Macaque (Macaca arctoides), etc. [Conclusion] This study, for the first time, used video monitoring technology to assess the natural enemies of endangered pheasant nests and clarified the behavioral patterns of natural enemies’ predation. Meanwhile, through literature review, we speculated that the longer incubation period and departure time might be the reason for the high rate of egg loss in T. caboti nests. This work is of great significance to the conservation and breeding of T. caboti.

Abstract:[Objectives] Since 2007, five released populations of Crested Ibis (Nipponia nippon) have been established in Ningshan, Tongchuan and Qianyang in Shaanxi Province, as well as Dongzhai in Henan Province and Deqing in Zhejiang Province. The reproductive behavior is important to the life history strategies, and the study on the reproductive ecology of the reintroduced Crested Ibis in Deqing can also reflect the nutritional status and local habitat quality of Crested Ibis as well as their adaptation to southern China. This paper can provide scientific support for the establishment of Crested Ibis population in southern China. [Methods] The reproductive behavior of released Crested Ibis in Deqing, Zhejiang Province, was recorded by video cameras for a total of 134 d and valid data for 2 958 h during the incubation and brooding stages in April-May 2018 and March-May 2020 in Deqing Xiazhu Lake National Wetland Park.The linear mixed models were used to explore the influence factors of the frequency of supplying nest-material and the reproductive behavior of Crested Ibis. We used Spearman correlation analysis to examine the relationship between the egg-turning frequency and the incubation process. The logistic regression model was used to explore the change of fledgling-warming duration of parent birds with the day age of fledglings. The one-way ANOVA was used to analyze the differences about nest-changing frequency between Deqing population and wild population as well as the other reintroduced populations. [Results] The results showed that the mean clutch size of the reintroduced Crested Ibis in Deqing was 3.7 ± 0.3. The hatching success rate was 90.9% and the fledgling success rate was 100.0% (Table 1). Different breeding periods, nesting conditions and the gender of parent birds significantly affected the frequency of supplying nest-material of Crested Ibis (Table 2, 3). The egg-turning frequency decreased significantly with the incubation process (Table 4, 5). The largest decline of the duration of chick-warming was at 11 days of age (Fig.1). Different brooding periods had a significant impact on the duration of fledgling-warming and the frequency of feeding and nest-changing in the Crested Ibis (Table 6, 7). [Conclusion] The higher environment temperature may be one of the reasons why the population of Deqing Crested Ibis lied eggs earlier than wild population in Yangxian County. The higher average clutch size and fledgling success rate may indicate that the Crested Ibis was adapted to the habitat of southern China. The Crested Ibis which used artificial nest-baskets spent more time supplying nest-material, indicating the necessity to improve the artificial nest-basket design in the future. We suggest to increase the open angle and reduce the depth of the nest-basket, and expand the outer diameter that close to the natural nests. At the same time, the bottom of the nest-basket should be designed as a porous and breathable structure that is conducive to drainage, in order to adapt to the humid and rainy climate in the middle and lower reaches of the Yangtze River in China.

Abstract:[Objectives] It is well known that human disturbance has been affecting the habitat, survival, reproduction of wild animals, with the negative impacts depending on the intensity of human disturbance. In order to understand how the human disturbance impacts wild animals, we set up 40 infrared cameras at the previously selected sites with distinct disturbance intensities in Baishanzu National Nature Reserve, Zhejiang Province, China in January 2021. [Methods] Based on the differences in the human disturbance intensity, the selected sites were divided into four groups (i.e., few disturbance area, slight disturbance area, minor disturbance area and serious disturbance area, from low to high human disturbance level). The monitoring period lasted 365 days, and there were 14 585 camera days in total.Shapiro-test of R software was used for normality test. t-test was used to analyze the difference if the normal distribution was met, and Wilcox-test was used to analyze the difference if the distribution of data was not normal. [Results] 4 256 independent photographs were taken, of which 3 485 were for mammals and 771 were for birds. We found there were 31 species of animals photographed besides rodents, including 14 mammals and 17 birds. Specifically, two of those were under the first-class state protection and nine belonged to the second-class state protection. Because of the blurry images, three photographs of birds were not identified to species. The number of animal species photographed in few disturbance area, slight disturbance area, minor disturbance area and serious disturbance area were 29 (15 mammals and 14 birds with 1 798 photographs), 24 (15 mammals and 9 birds with 1 308 photographs), 20 (12 mammals and 8 birds with 756 photographs) and 18 (13 mammals and 5 birds with 394 photographs), respectively. It was found that there was a significantly negative relationship between the degree of human disturbance and the number of independent photographs (P < 0.01). Namely, with the human disturbance intensifying, less species of animals, particularly for birds, were photographed. Moreover, 3 163 of these photographs were taken from 23 infrared cameras deployed in the state-owned forests, being identified to 16 mammals and 17 birds. In the collective-owned forests, 17 infrared cameras produced 1 093 photographs (identified to 13 mammals and 9 birds). The number of photographs taken by one camera in the collective-owned forests was only 46.75% of that in the state-owned forests, with a significant difference between them (P < 0.01), possibly due to forest cutting in the early years and thus the destroyed and fragmented habitat had a negative impact on animals. [Conclusion] As such, habitat protection should be an important task for wildlife conservation and be taken seriously.

Abstract:[Objectives] Jiangcheng is located in the priority area of biodiversity conservation in Xishuangbanna, where the diversity of reptiles is unclear and reptile resource survey is lack in history. Here, we conducted seven surveys for reptile diversity in Jiangcheng from May 2019 to September 2020. [Methods] Comprehensive surveys have been conducted to evaluate diversity of reptiles by using the fixed line transects method. A total of 207 survey lines were set up, covering a wide range of diverse habitats at different altitudes. The time span of each day is from 8:00 am to about 04:00 am of the next day. Collected specimens were brought back to the laboratory for identification. We use GrcGIS and Excel for data mapping and statistics. For data analysis, we used Berger-parker index (I) to determine dominant species, Margalef index (DMg) to determine species richness, and G-F index (DG-F) to analyze family and genus diversity. [Results] A total of 51 species of reptiles were recorded in Jiangcheng, which belong to 2 orders, 17 families, and 39 genera. 17 species (33.3%) are endangered in China and one Class-I (1.9%) and 7 Class-II (13.7%) nationally protected reptiles were recorded. The dominant species are Hemidactylus bowringii (I = 13.3%), Pareas macularius (I = 11%), while the rare species are Platysternon megacephalum, Cyclemys oldhami, Varanus salvator and Python bivittatus in Jiangcheng. We found 2 newly identified species which are namely Pareas geminates and Trimeresurus goui. Two species were new records. One of them, Parafimbrios lao, is the new record in China and the other, Pseudoxenodon karlschmidti, is the new record in Yunnan Province. All species can be divided into 6 ecological types, which are residential (3.92%), arboreal (17.65%), bushwood rocky (45.10%), arboreal aquatic (23.53%), aquatic (5.88%) and soil (3.92%, Fig. 2). Form the taxon of all, the Testudofomes have high dependence on the aquatic ecological environment, and the Lacertilia and Serpentes have a high dependence on the forest ecological environment. The soil and the residential type are endemic to the Serpentes and Lacertilia, respectively. The bushwood rocky has the highest richness index (DMg = 3.548) with high species evenness and the best stability. On the contrary, residential (DMg = 0.198) is the lowest richness index with low species evenness and poor stability (Table 1). The vertical distribution shows a certain regularity. The mountain slope (700﹣1 500 m) has the most abundant species, with 47 species whereas the Piedmont ravine and the mountain top are less distributed, with 10 and 4 species (Fig. 3). There are many species of single genus in reptiles (G = 3.506 5, F = 8.406 0). Species are mainly in the Oriental boundary, and has a relationship with the Himalayas-Hengduan Mountains. According to this survey, road killing is the most direct factor leading to the threat of reptiles, and 2/3 of the survey data comes from road killing, involving 35 species, especially Pareas macularius and Bungarus wanghaotingi. In addition, overharvesting has made some economically valuable species become rare, such as turtles and pythons. [Conclusion] Jiangcheng is rich in reptile species diversity, and this survey makes up for the gap in the literature on reptile diversity in this area. Jiangcheng is also a typical area of the biodiversity hotspot for studying the reptile diversity of rainforest. We suggest that the existing forest distribution pattern should be maintained, as well as keep studying, monitoring and conservation of reptile diversity into the future.

Abstract:[Objectives] Small and medium-sized soil animals are important components of soil fauna in terrestrial ecosystems. Research shows that there are many biotic and abiotic factors that affect soil fauna diversity in anthropogenic ecosystems such as paddy fields, and anthropogenic disturbance factors such as planting methods play the most obvious role. Chongming Island is an important agricultural planting area in Shanghai. The influence on ecosystem of rice farming is the focus in the rice production process. At present, although there have been some reports on the community structure and diversity characteristics of soil animals in paddy fields under different planting methods, these studies have not yet covered the three rice planting methods of conventional, organic and ecological (rice-fishing) in the same area. Therefore, the present study investigated the differences in the diversity of small and medium-sized soil animals under three rice cultivation methods. [Methods] This study set up three sample plots, which are all located on Chongming Island. The three plots are mainly cultivated with rice, and their natural vegetation and production background are basically the same. Their farming patterns were conventional planting, organic planting and ecological planting (Table 1). According to different rice growth stages, this study investigated the small and medium-sized soil animals in paddy fields in January, March, June and October, 2020, with a total of 4 samples. Small and medium-sized soil animals were separated by Tullgren method, and one-way ANOVA and Kruskal-Wallis H method were used to test the significance of differences. [Results] The results showed that a total of 449 small and medium-sized soil animals were obtained, belonging to 2 phyla, 5 classes, and 8 orders (Table 2). The dominant group was Collembola, accounting for 82.18% of the total catch. Among the three planting methods, Symphyla are only found in the soil of conventional paddy fields, Rosette is only found in the soil of organic paddy fields, and Hemiptera and Orthoptera are only found in the soil of ecological paddy fields. The community structure of small and medium-sized soil animals were significantly different in paddy fields under three planting methods, the density difference of small and medium-sized soil animals in different plots were significant, and the density of organic paddy fields was higher than the others (Fig. 1). The vertical distribution of small and medium-sized soil animal communities showed surface clustering characteristics, and the number and density of groups showed a downward trend (Fig. 2). There was no significant difference in the number of groups of small and medium-sized soil animals in different soil plots. The richness index, diversity index, and evenness index of small and medium-sized soil animals in ecological planting paddy fields were the highest, and the dominance index of organic planting paddy fields was the highest (Fig. 3). Three different planting patterns showed moderate similarity between any two groups. [Conclusion] Compared with conventional and organic planting patterns, ecological planting pattern is more conducive to improving the diversity and evenness of small and medium-sized soil animals and promoting the stability and healthy development of the paddy ecosystem. It is of significance to the sustainability of rice planting and as the indicator of agricultural non-point source pollution. At the same time, there are many factors affecting the community structure of small and medium-sized soil fauna in paddy fields, we suggest that further studies can be performed with the correlation among climate factors, physical and chemical indexes, and soil fauna community.

Abstract:[Objectives] During the 14^th Five-Year Plan period, the Pinglu Canal in Guangxi will be a magnificent project for New International Land-Sea Trade Corridor in western China, and the Maowei Sea is a critical point of the Canal. Maoling River and Qin River flow toward the Maowei Sea. Land-sourced pollutants are inputted by these two rivers. The pollutants mainly include industrial wastewater and domestic sewage, as well as pesticides and fertilizers in agricultural irrigation areas, and nutrients and organic pollutants in surrounding shrimp-breeding pond. From 2010 to 2014, the water quality of the Maowei Sea was the worst, with pollution readings often reaching level four. The main factors that exceeded the state standard were inorganic nitrogen and active phosphate. The pH of some part of Maowei Sea also exceeded the state standard. Because the outlet to sea is narrow, and too much pollutants inputted, the routine tides from Beibu Gulf cannot effectively keep the water quality of the Maowei Sea at a healthy level. The community of macrobenthos in the Maowei Sea in 2009 was investigated, and it was believed that the Maowei Sea was in a clean to lightly polluted situation. Therefore, the present study aims to assess the ecological environmental level of the Maowei Sea using intertidal macrobenthic communities, and to collect the baseline data which would support the scientific management of the Pinglu Canal during construction and operation. [Methods] Intertidal macrobenthos were collected at four sampling sites along the eastern coastline of Maowei Sea, Qinzhou City, which included Sun Yat-Sen Park, Shahuan Village, Mangrove Park and Kangwangmiao Village, in October 2020 as well as in January, April and July 2021 (Fig. 1), according to the standard protocol for marine studies (GB 12763.6-2007). One sample line perpendicular to the shoreline is set for each site, and one sample is taken in the high tide zone, two samples are taken in the middle tide zone, and one sample is taken in the low tide zone. Then all animals from all samples mixed together, which represent the community of macrobenthos at the sampling site. Samplings were conducted once every season in every sample site for one year. All sites are outside mangroves or on bare beaches. The mud samples were filtered through sieves (mesh size:0.5 mm) in the nearby tidal creeks. The macrobenthos samples in sediments were fixed in 5% formalin in the field. All specimens of macrobenthos were brought back to the laboratory. The collected macrobenthos were preserved in 75% ethanol for further examination in the laboratory. All specimens were identified by ourselves and some taxonomy experts. All specimens were weighed by electronic balance at 0.01 g accuracy. All specimens were counted and categorized to clarify the macrobenthic composition of each sampling site. Index of relative importance (IRI) was calculated to clarify dominant species, main species, common species, and rare species. IRI = (pi + wi) fi, where pi is the relative abundance of species i in the plot, wi is the proportion of the weight of species i in the plot (%), and fi is the percentage of the number of plot of species i in the total number of plots investigated (%).We defined IRI≥ 1000 as dominant species, 100 ≤ IRI < 1000 as main species, 10 ≤ IRI < 100 as common species, and IRI < 10 as rare species. Species with IRI ≥ 100 were considered as the important species in the fauna community. Simpson diversity index (D), Shannon diversity index (H?), Pielou evenness index (J) are used to scale the diversity of macrobenthos. Nonparametric Kruskal-Wallis and Mann-Whitney analyses were performed on the macrobenthos biodiversity index data using SPSS 26.0. Then diversity of macrobenthic data was analyzed and compared to assess the overall environmental conditions of the Maowei Sea. Shannon’s diversity index H? is used to assess environmental pollution, while H? < 1, heavy pollution; H? value of 1 to 2, moderate pollution; H? value of 2 to 3, mild pollution; H? > 3, clean. Meanwhile, macrozoobenthos pollution index (IMP) assesses pollution level, while IMP <﹣6, clean;﹣6 ≤ IMP < 0, light pollution; 0 ≤ IMP ≤ 4, moderate pollution; IMP > 4, serious pollution. The multivariate analysis software PRIMER 6.0 was used to analyze the disturbance degree of macrobenthos community, and the abundance and biomass curve (ABC) was constructed. When the biomass dominance curve is above the abundance curve, W is positive, which means the community is not disturbed; on the contrary; when W is negative, it means that the community is severely disturbed and the biomass curve is below the abundance curve; When the curves overlap, the W value is close to 0, indicating moderate interference. [Results] Total of 3 084 macrobenthos individuals were collected from 4 sites in the intertidal zone of the Maowei Sea. According to the classification and identification, there are 6 phyla, 8 classes, 31 families, 51 genera and 58 species of macrobenthos in the Maowei Sea intertidal zone (Table 1). The average annual animal density is highest in Shahuan Village (6 340 ind/m²); and lower in Sun yat-sen Park (1 240 ind/m²), Mangrove Park (3 204 ind/m²)and Kangwangmiao Village (1 552 ind/m²). The macrobenthos composition in each sampling site is more than 50% arthropods, followed by annelids, about 10%, and molluscs, less than 2%. Small arthropods are the main group of macrobenthos in the intertidal zone of the Maowei Sea, accounting for 41.38% of the total species and 87.13% of the total individuals. However, the macrobenthos communities in the estuaries and intertidal zones of the Beibu Gulf in the past, were generally composed of molluscs, arthropods, and annelids in turn as the dominant groups. To date (Table 2), Discapseudes mackiei, Ilyoplax formosensis, Macrophthalmus convexus, Metopograpsusquadridentatus whose IRI are more than 1000, are the dominant species, but dominant species were not evenly distributed in the four plots, and only D. mackiei appeared in each plot. 27 main species whose IRI are more than 100 but less than 1 000 are listed in Table 2, Paracleistostoma depressum, Notodasus sp. appeared in each plot, however, the others were not evenly distributed in plots. 19 common species whose IRI are more than 10 but less than 100 were found, and 8 rare species whose IRI are less than 10 were found. According to Kruskal-Wallis analysis, the Simpson diversity index (D) and Shannon diversity index (H?) values of Shahuan Village were all lower than other three sites (Table 3). The average indices of the Simpson diversity index (D) and Shannon diversity index (H?) values were 0.435 (range 0.183﹣0.704), and 1.506 (range 0.747﹣2.256) respectively, in which the levels were significantly lower than those in 2009 (P = 0.004, and P = 0.048 for D and H? respectively). The Pielou evenness index (J) value was 0.475 (range 0.223﹣0.696) which was statistically similar to that in 2009 (P = 0.109). Overall consideration of abundance and biomass curve (ABC) (Fig. 2), Shannon diversity index (H?) and macrozoobenthos pollution index (IMP), environmental quality of the Maowei Sea is in moderate pollution. [Conclusion] Our results demonstrated that the intertidal areas of Maowei Sea were dominated by arthropods, which accounted for over 41% of the total species number and over 87% of the total abundance. The dominant species (IRI > 1 000) included D.mackiei, I.formosensis, M.convexus and M.quadridentatus, but they were not evenly distributed in the four sites, and only D. mackiei appeared in each site. These biodiversity indices concluded that the Maowei Sea is suffered from moderate pollution. The currently established intertidal macrobenthic database of Maowei Sea can support the ecological construction and scientific operation of the Pinglu Canal in future.

Abstract:[Objectives] Positive reinforcement blood collection training allows to collect blood samples from animals without physical or chemical bonding, minimizing injuries due to stress. In conventional positive reinforcement blood collection training, non-fixed reinforcement, mainly food, is used as a reinforcer to maintain the movement. In this study, after failing to train three chimpanzees in blood collection using non-fixed reinforcement, music was added to the blood collection training as a continuous positive reinforce and timer for the first time, based on the characteristic that chimpanzees love music and can recognize musical melodies. [Methods] The retention time of blood collection action of each chimpanzee during training without music and with music was recorded respectively. SPSS Statistics (version 20.0) was used for statistical analysis. First, Shapior-Wilk test was used for normality test. For data conforming to normal distribution, paired sample t-test was used for testing. Conversely, Wilcoxon signed-rank test was used for data that are not normally distributed. [Results] When using music for blood collection training, the movement retention time of the three chimpanzees was significantly higher than that of the non-fixed reinforcement method (without music) (Table 1). The number of times each chimpanzee completed the training, that is, the minimun number of times required to achieve the required training effect varies. Beibei had the most training sessions (177 times) and the training difficulty was sharp needle desensitization for 4 min, followed by Yaya (69 times) and the training difficulty was holding the movement for 4 min. Mingming had the fewest training sessions (42 times) (Table 2). [Conclusion] Upon completion of training, all three chimpanzees were able to complete blood collection during 4 min of music playing, without feeding any food in the middle of the session. That is, music can prolong the movement retention time of chimpanzees during blood collection training and increase the stability of training. The methods and study can provide a sound reference and approach for related workers in the same industry.

Abstract:[Objectives] The study aims to explore the effects of exogenous gonadotropin on epidermal growth factor (EGF) expression level and action characteristics in Hyla Rabbit’s (Oryctolagus cuniculu) reproductive organs. [Methods] In this experiment, 30 Hyla Rabbits (6 male and 24 female) were randomly selected. Female rabbits were randomly divided into superovulation group (intramuscular injection of 70 U pregnant mare serum gonadotropin +100 U human chorionic gonadotropin, n = 12) and control group (no injection of any hormone, n = 12). And Real-time fluorescence quantitative PCR (qRT-PCR), Western-blot and immunohistochemistry were used to study the expression and localization of epidermal growth factor (EGF) in ovaries, oviduct and uterus in superovulation group. SPSS 25.0 software was used for independent sample t test analysis of relevant data. Results were expressed as “mean ± standard deviation”, P < 0.01 means significant difference, P < 0.05 means significant difference. [Results] The results showed that the relative expression level of epidermal growth factor mRNA in different tissues was significantly higher in the superovulation group than in the control group (uterus:t = 17.053, P < 0.01; oviduct:t = 3.009, P < 0.01; ovaries:t = 14.125, P < 0.01) (Fig.1). The relative expression of EGF protein was also significantly higher in different tissues of the superovulation group than the control group (uterus:t = 20.663, P < 0.01; oviduct:t = 19.537, P < 0.01; ovaries:t = 21.272, P < 0.01) (Fig.2). And the positive signals were located in primary oocytes, granulosa cells, endometrium cells, mesenchymal cells and vascular endothelial cells respectively (Fig. 3). Oviduct ciliated cells, basal cells, mucosal epithelium, muscularis and epithelial (Fig. 4), and endometrial cells of the uterus (Fig. 5). [Conclusion] Our study has shown that exogenous gonadotropin used for superovulation can promote the expression of epidermal growth factor expression in ovaries, fallopian tubes and uterus, suggesting that EGF may be involved in the reproductive process of Hyla Rabbit and regulate the functions of ovaries, fallopian tubes and uterus.

Abstract:[Objectives] Research on oxidative stress in skeletal muscle under disuse conditions is a topic of increasing concern among physiologists. In view of the important role of elevated oxidative stress in the development of disuse muscle atrophy, we hypothesized that hibernators and non-hibernators may exhibit differences in oxidative stress levels and antioxidant defense regulation, thus constituting intrinsic differences in the mechanism of muscle atrophy. Moreover, because of the variations in their energy metabolism and function, differences should also exist in the degree of atrophy and level of oxidative stress in different skeletal muscles. For this reason, we investigated the degree of atrophy in the SOL (slow-twitch) and EDL (fast-twitch) muscles in parallel with the level of oxidative stress, antioxidant defense and underlying mechanism in non-hibernators (Rats, Rattus norvegicus) and hibernators (Daurian Ground Squirrels, Spermophilus dauricus) between artificial and natural disuse. [Methods] We examined changes in muscle wet weight and muscle-to-body weight ratio by electronic balance. Immunofluorescence staining was performed to measure the muscle fiber cross-sectional area (CSA) and fiber type composition. Using a hydrogen peroxide (H2O2) assay kit and malondialdehyde (MDA) assay kit, we measured the concentrations of H2O2 and MDA (level of oxidative stress) in muscle samples. The antioxidant activity was ascertained using specific superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) assay kits. We detected protein expression of the Nrf2/Keap1 signaling pathway and downstream antioxidant enzymes (SOD1, SOD2, CAT, and GPx) in hindlimb loading (HU) Rats and hibernating Daurian Ground Squirrels. Differences in body weight between HU and control Rats and between hibernating inactive (HIB) and summer active (SA) Ground Squirrels were evaluated with an independent samples t-test. Muscle wet weight, ratio of muscle wet weight/body weight, fiber-type distribution, fiber CSA, and all the protein expression levels were analyzed using two-way ANOVA (main effects of muscle type and disuse treatment) followed by independent samples t-test. [Results] Compared to the control Rats, muscle atrophy indices (muscle mass, muscle-to-body mass ratio and CSA) were significantly increased in the SOL and EDL after 14 d of HU. In the Daurian Ground Squirrels, however, no significant changes in these indices were observed in the muscles between the hibernation inactive and summer active groups (Fig. 1 and 2). After 14 d of HU, the proportion of fast type fiber (MyHC II) increased significantly by 206.0% (P < 0.001) in the SOL compared with the control Rats; however, there was no significant change in the proportion of fiber type in the EDL. In Daurian Ground Squirrels, the proportion of slow type fiber (MyHC I) and MyHC II showed no significant changes in the SOL or EDL between the hibernation inactive and summer active groups (Fig. 2). In regard to oxidative stress, H2O2 content increased significantly in the SOL and EDL and MDA content increased significantly in the SOL of HU rats; however, neither were increased in the hibernating Daurian Ground Squirrels, although H2O2 and MDA content decreased significantly in the EDL (Fig. 3). SOD2, CAT, and GPx1 expression markedly decreased in the SOL, as did GPx1 in the EDL, whereas SOD1 expression increased significantly in the SOL of HU rats. In hibernating Daurian Ground Squirrels, however, expression was maintained or increased in both muscles; for example, CAT and GPx1 protein levels increased significantly in the SOL and EDL of the hibernation group (Fig. 5 and 6). Furthermore, compared with control rats, Nrf2, p-Nrf2, and Keap1 protein levels in the SOL of the HU group demonstrated no changes, whereas the expression of Nrf2 and Keap1 in the EDL of the HU group decreased significantly, whereas Nrf2 and p-Nrf2 protein levels in the SOL and EDL of the hibernating Daurian Ground Squirrels increased markedly (Fig. 7). [Conclusion] These results indicate that the Nrf2/Keap1 antioxidant pathway was not activated and downstream antioxidant enzymes were down-regulated in Rats under muscle disuse conditions, which may be an important mechanism in oxidative stress (i.e., increased reactive oxygen species) and induced skeletal muscle atrophy. Conversely, activation of the Nrf2/Keap1 signaling pathway and increase in antioxidant enzymes observed in the Daurian Ground Squirrels during hibernation undoubtedly helped prevent oxidative stress and muscle atrophy. In conclusion, oxidative stress and Nrf2-mediated antioxidant defense appear to differ in skeletal muscles of non-hibernators (Rats) and hibernators (Daurian Ground Squirrels) under disuse conditions, which may be important mechanisms for the differing degrees of muscle atrophy.

Abstract:[Objectives] Chinese Soft-shelled Turtle (Pelodiscus sinensis) is an important aquatic economic animal, belonging to the representative taxonomic family of Trionychidae. The glans of the penis of P. sinensis is the black part of the end of the penis, which is the key to the study of mating and sperm excretion of the turtle. At present, its detailed structure has not been reported. [Methods] In this study, six-year-old sexually mature male P. sinensis were used. The penile glans of P. sinensis were studied in erectile and natural state by anatomical and histological methods (stereomicroscope, light microscope and scanning electron microscope). [Results] The results showed that there were two sulcus spermaticus on the dorsal side of the penis (Fig. 1d), which extended along the penis to the synaptic tissue of the penis head. The dorsal surface of the penis head is a fold like blood sinus, which expands after the congestion of the penis. When the blood sinus fully extended, ten tentacles were appeared on the surface of penis glans, and formed a symmetric “butterfly” structure, which can open and close during ejaculation. In addition, the head of the penis is rich in villi and mucus. [Conclusion] The penile glans of P. sinensis formed a special structure in the erectile state, which may be directly related to the stability of mating and spermatogenesis.

Abstract:[Objectives] The olfactory and vomeronasal organs are present and well developed in squamates (lizards and snakes). To elucidate the evolution of chemoreception in reptiles, a detailed structural examination of the vomeronasal organs from a variety of squamate species is required. [Methods] The histological features of the nasal cavity and vomeronasal organ in Phrynocephalus frontalis,P. przewalskii, Eremias multiocellata and Scincella tsinlingensis wereexamined using hematoxylin-eosin staining. The data was analyzed using one-way Analysis of Variance with Duncan’s method for multiple comparisons between species. [Results] The nasal cavity of P. frontalis is long and narrow, while the nasal cavity of S. tsinlingensis is pear-shaped; in contrast, the nasal cavity of other two lizards are slightly round (Fig. 1). The olfactory epithelium of S. tsinlingensis is the thickest, followed by E. multiocellata and P. frontalis, and the olfactory epithelium of P. przewalskii is the thinnest (P < 0.05) (Fig. 2 and 3). The vomeronasal organ is composed of the vomeronasal lumen, vomeronasal sensory epithelium, vomeronasal nerve and mushroom body. No glandular structures were found in the vomeronasal organ. The vomeronasal lumens of P. frontalis and P. przewalskii are broader than E. multiocellata and S. tsinlingensis. The vomeronasal sensory epithelium is thicker than the olfactory epithelium in four species of lizards. At the caudal portion of the vomeronasal organ, the mushroom body disappears and the vomeronasal sensory epithelium entirely lines the circular lumen of the vomeronasal organ. The vomeronasal sensory epithelium is situated dorsally to the vomeronasal organ in E. multiocellata and S. tsinlingensis, while the mushroom body occupies the veutri of the vomeronasal organ; in contrast, the vomeronasal sensory epithelium is situated ventrally to the vomeronasal organ in P. frontalis and P. przewalskii, while the mushroom body occupies the dorsum of the vomeronasal organ (Fig. 1 and 4). At the middle region of the vomeronasal organ, the vomeronasal sensory epithelium of E. multiocellata is the thickest, followed by S. tsinlingensis, and the vomeronasal sensory epithelium of two species of Phrynocephalus is thinner. The density of vomeronasal sensory cells of S. tsinlingensis is the highest, followed by E. multiocellata, and the two species of Phrynocephalus are the lowest (Fig. 3). [Conclusion] These results indicate that E. multiocellata and S. tsinlingensis use and invest more in chemical signal than two species of Phrynocephalus. The differential features of the vomeronasal organ from four species of lizards indirectly confirm the specificity of vomeronasal organ phylogeny in squamates.

Abstract:In August 2021, we collected one specimen of the genus Hebius (Fig. 1 showing the morphological characteristics of this snake) in the Dabie Mountains (Niuwang Mountain, Lujiang County, Hefei City, Anhui Province, China). Morphological comparison showed that the snake was obviously different from the recorded genus Hebius in the Dabie Mountains (H. sauteri and H. craspedogaster). Based on molecular phylogenetic analysis, it was shown that the specimen was clustered with H. vibakari and morphologically, it conforms to the characteristics of H. vibakari (Fig. 2). It suggests that the specimen should be H. vibakari. This is a new recorded species of H. vibakari distribution in the Dabie Mountains and Anhui Province, and this is the first time of this species reported outside of northeastern China. This new record expands the distribution range of H. vibakari, and is of great significance for studying the population evolution and biogeography of this species.

Abstract:Two species of barbastelle live in China, Barbastellabeijingensis and B. darjelingensis. B. darjelingensis is found in Inner Mongolia, Xinjiang, Qinghai, Shaanxi, Gansu, Henan, Hunan, Jiangxi, Sichuan, Chongqing, Yunnan, Guizhou, and Taiwan, while B.beijingensis has only been recorded in Beijing. A male barbastelle was captured by mist net in the forest road of Changyuanhe Wetland Park in Qi County (37°19′15″ N, 112°26′18″ E, 812 m), Shanxi Province, in July 2022. We measured the external morphology and skull characteristics of the captured sample, and amplified the Cyt b and ND1 sequences. With 1 000 bootstrap replicates, phylogenetic trees were reconstructed from Cyt b and ND1 gene sequences by Mega 5.0. The forearm length is 43.7 mm, and the sample had nearly square outline of ears with transverse ridges (Fig. 1). Its ears joined across the forehead, and it had a small lobe on the middle outer edge of each pinna (Fig. 1b, c). This Shanxi sample had a longer 3^rd, a middle 4^th and a shorter 5^th metacarpal lengths (Table 1). The length of skull is 14.6 mm, and it has robust upper and lower premolars (Fig. 2). All morphological measurements were in accord with those from Barbastella beijingensis, except condylo-base length and braincase breadth (Table 1). Cyt b and ND1 sequences from the Shanxi sample showed higher similarities to those of B. beijingensis (Cyt b:99.5%; ND1:98.6% to 100%), than to those of B. darjelingensis (Cyt b:86.3% to 86.9%; ND1:81.6% to 83.3%). The Shanxi sample clustered with B. beijingensis, rather than B. darjelingensis in both cladograms of Cyt b and ND1 genes(Fig. 3). Through comparing external morphology, skull characteristics and molecular data with B. beijingensis and B. darjelingensis, this bat is identified as B. beijingensis, which is a new record for the Chiroptera in Shanxi Province.

Abstract:In June 2022, a single specimen of amphibian was collected from Hualongshan Nature Reserve, Zhenping County, Ankang City, Shaanxi Province (32°00′01″ N, 109°17′05″ E, altitude 1 641 m), and identified as a species of the genus Odorrana by comparison of morphological characters. In the phylogeny of 35 species of the genus Odorrana based on the mitochondrial 16SrRNA gene, it clustered with O. kuangwuensis, which was distributed in Nanjiang, Sichuan, the type locality, in the ML tree with 99% support, and should be intraspecific. Based on the morphological analysis and phylogenetic comparison, the collected specimen was identified as O. kuangwuensis, a genus of frogs in the Anura family (Ranidae), which represents a new record of amphibians in Shaanxi Province, and provides evidence for the continuity of the distribution of O. kuangwuensis in the Daba Mountain Range.

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