Abstract:The Eurasian Lynx (Lynx lynx) is the top predator at Saihanwula Nature Reserve in Inner Mongolia, which functioned as the major controller on prey abundance and community composition for the regional wild animal ecological balance. Study on the fecal residue of the lynx could reveal the food components and understanding the prey sharing mechanism with other sympatric carnivores. Moreover, this baseline information will provide references for endangered species conservation and habitat management in the reserve. In this study we collected 35 lynx scats by line transact sampling within the nature reserve from 2006 to 2008, and the relative occurrence frequency index was used to compare the food components (Fig. 1). The hare (30.85%) and plants (28.72%) occurred mostly in the scats, while the frequency of each food items varied annual (χ2 = 18.696, P < 0.001) and seasonal (χ2 = 74.695, P < 0.001, Table 1 and 2). Hare played a crucial role in lynx food spectrum at the study area. The lynx is an opportunistic predator who captures on large size prey more often in the cold season while feeds on small size prey frequently in the warm season.

Abstract:The Cinereous Vulture (Aegypius monachus) is listed as a Near-Threatened species by IUCN. Each year in winter or early spring, a few weak or poisoned vultures were rescued in Bohai Bay area. In the spring of 2016, two rescued vultures (ID2016 and ID1051, basic information shown in Table 1) were equipped with satellite transmitter and their migratory routes and home ranges were analyzed, aiming to provide more effective conservation measures. The two vultures migrated to the border of Bulgan and Arkhangai Province of Mongolia, and travelled about 2 200﹣2 400 km from wintering to summering area (Fig. 1). The home range sizes (50% Fixed Kernel) in the early, middle and late summer of the ID2016 were 31 261.01 km2, 1 492.96 km2 and 11 027.70 km2 respectively (Fig. 2). During migration, the vultures spent a majority of their time in the process of resting, followed by flying and feeding behaviors. They normally started to move around 8:00, reached the activity peak around 12:00 to 13:00, then gradually slowed down until 18:00 to be stable. This study provided a complete spring migration routes and important stopover sites for the Cinereous Vultures, and it was the baseline for the rescuing and protection of the vultures.

Abstract:During breeding season, from May to August 2017, we collected songs from 80 breeding males of Dusky Warbler (Phylloscopus fuscatus) in Saihanba, Hebei Province. The changes of song types, song parameters and song syllable types were analyzed. There were two sentence patterns in the song of the males: S-song (Fig. 1a) and V-song (Fig. 1b). There were significant differences in the song output of S-song and V-song among months (ANCOVA: F3,75 = 17.983, P < 0.05; F3,75 = 17.983, P < 0.05; Fig. 3), with more S-song used than V-song in the whole breeding season. The output of S-song and V-song in each month showed the opposite trend, with relatively more output of S-song in early (May) and late (August) breeding season, and more output of V-song in the middle breeding season (June). There was significant difference in the maximum frequency (ANCOVA: F3,75 = 4.841, P < 0.05; Table 1), bandwidth (ANCOVA: F3,75 = 3.814, P < 0.05; Table 1) and peak frequency (ANCOVA: F3,75 = 3.793, P < 0.05; Table 1) among months. Based on the 2 400 song verses from 80 males, 65 syllable types were obtained (Fig. 4). More than 80% of the syllable types were used in all months (Fig. 5), but the number of syllable types used in each month were significantly varied (ANCOVA: F3,75 = 5.456, P < 0.05; Fig. 6). The peak of syllable diversity was in July, with 19.79 ± 3.69 syllable types used, there were fewer syllable types used in May and August, with the least in August.

Abstract:All species of swans (Cygnus spp.) have been listed as the second class national protected animals in China. Illegal hunting, catching and killing of precious and endangered wild animal constitutes a criminal offence under the Law in China, however, such cases were still often happened. We collected and analyzed 46 death cases of swans during 2000 to 2016 in China. Our results showed that such cases were occurred frequently in winter from November to January. Poisoning, such as using the carbofuran, was the most important way to cause the deaths of swans. The cases always happened in borders between cities in China (Fig. 1). Therefore, pesticide management should be strengthened during the migration season of swans, especially in border areas between provinces and cities. Furthermore, to establish a multi-area law enforcement linkage mechanism during the migration season may be useful for protecting the swans.

Abstract:In China, gas cannons have been commonly applied in the airports to repel birds, by the high decibel noise generated by the gas explosion. However, the effectiveness of bird repelling varied due to long-term exposure to the noise, or unsuitable deployment time of the Gas cannon, making birds subjective to be habituated and even insensitive to noise. Quantitative evaluation of the effectiveness of gas cannons to repel birds is still scarce. In this study, adult and healthy domestic pigeons (Columba livia domestica) were used as the research object. 40 pigeons were randomly divided into eight groups, one non-treated control group, aiming to investigate the effects of background noise on corticosterone changes, and seven interference groups with a 20 m distance interval to the noise source. Fecal samples were collected the day before noise treatment, and then the pigeons were treated with gas cannon noise continuously with a frequency of 4 times/h for 20 days, during which samples were collected at day 2, 5, 8, 11, 14, 17 and 20. Fecal corticosterone was measured using ELISA. The fecal corticosterone concentration was used as the dependent variable, and the noise duration and the distance to the noise source as independent variables. The correlation between the two variables was analyzed using Pearson correlation analysis. Difference analysis was performed using one-way analysis of variance (ANOVA). The maximum corticosterone content was 174.67 ± 5.09 ng/g at 10 m, and a minimum of 139.42 ± 0.92 ng/g appeared at 130 m (Fig. 2). There was a significant negative correlation between the distant to noise resources and the corticosterone content in the feces (Pearson′s r =﹣0.954, P < 0.01) (Fig. 1). The noise duration and the distance to the noise source have a significant interaction effect on the fecal corticosterone (F = 15.008, P < 0.01) (Table 1). As the distance increased, duration time for induced corticosterone back to the baseline showed a decreasing, and corticosterone did not change significantly with the time span beyond the noise distance of 90 m (Fig. 2). Fecal corticosterone could be an effective indicator to reflect the stress of birds to noise, and the effectiveness of gas cannons was evaluated, providing a scientific basis for control and prevention of bird strikes at the airport.

Abstract:In order to understand the male calls characteristics of the poorly known Leptobrachium tengchongense, field survey was conducted on 27th February 2018 in Dahetou Ranger Station of Gaoligongshan National Nature Reserve, Tengchong City of Yunnan, China, aiming to obtain the records of male calls during its breeding season. A total of 78 male calls, belonging to six adult male individuals, were recorded using a digital recorder (TASCAM DR-40); the calls were analyzed using the Raven Pro ? v.1.5 software. The results show that, adult males of Leptobrachium tengchongense could make two different types of calls during the breeding season, namely advertisement call and combat call (Fig. 1). Both are single-note calls, and all have a dominant frequency of 1.31﹣1.50 kHz (Table 1). The combat calls are normally emitted when there are other males calling nearby, and its call duration is comparatively longer than those of advertisement calls (610 ± 187.0 ms vs. 147 ± 30.1 ms), and contains more pulses (28 ± 10.7 pulses/s vs. 15 ±2.4 pulses/s). There are variations on the call duration and intercall-interval among six individuals, and these variations may correlate with body size; however, due to the limited sample size, more records are needed in order to clarify the relationship between the body size and its call characteristics. Based on the survey result, we also discuss the current two major threats, habitat degradation and human consumption on tadpoles, to the species and propose relevant conservation actions.

Abstract:Immunohistochemical SABC was performed to analyze the expression of ZENK protein in the brain of female concave-eared torrent frogs (Odorrana tomorta) under background noise, the voice of male concave-eared torrent frogs and the voice of the male large odorous frog (O. graminea). The results showed that there were no ZENK protein-positive cells in the telencephalon, diencephalon and cerebellum of the female concave-eared torrent frogs (Fig. 2, 4) stimulated by aforementioned three voices. The cells in optic tectum, interpeduncular nucleus rostral portion, nucleus profundus mesencephali, tegmentum, torus semicircularis of the mesencephalon and the cells in solit tract nucleus, raphe nuclei, reticular nuclei, reticular formation, superior olivary nucleus of the medulla oblongata in female concave-eared torrent frogs showed positive reaction under the stimulation by the voice of male concave-eared torrent frogs (Fig. 3, 5), while the numbers of positive cells in aforementioned areas of the brain were markedly reduced under the stimulation from the voice of the large odorous frog. Part of the ZENK protein-positive cells were located in the torus semicircularis, tegmentum of mesencephalon and the superior olive nucleus of medulla oblongata, which were related to the processing of auditory information in amphibians. It can be inferred that ZENK protein is closely related to the processing of auditory information in female concave-eared torrent frogs.

Abstract:We have sampled 194 Schizothorax nukiangensis, 152 Ptychobarbus kaznakovi and 117 Schizopygopsis thermalis in Tibet section of the Nujiang River between May and October 2017 (Fig. 1, 2; Table 2) in order to compare the morphological characteristics of digestive organs including feeding and digestive organs among the three Schizothoracinae fish species. Three methods of multivariate analysis were applied to analyze the morphological difference of feeding and digestive organs. The results showed that major parts of interspecific variation were the cephalic and enteral characteristics (Fig. 3, 4; Table 5, 6), and the morphological indexes of different species were significantly different among the three species (P < 0.05) (Table 3). The cephalic morphology of the three species was with differences. Analysis on the enteral inclusion under microscopes was applied to understand their food competition. The results indicated that Schizothorax nukiangensis and Ptychobarbus kaznakovi belonged to the omnivore and partial carnivore fish (Fig. 6), and the Schoener overlap index between two species was high (0.91). Schizopygopsis thermalis belonged to the omnivore and partial herbivore fish. The indexes of diversity of dietary composition among three species of fishes were apparently different (Table 8). Feeding habits of the three fishes were closely related to environmental factors, such as water temperature, flow rate and altitude (Table 1).

Abstract:The microstructure and ultrastructure of zebrafish (Danio rerio) spleen were observed by light microscopy and transmission electron microscopy. The results showed that the spleen of zebrafish mainly consisted of splenic pulp and reticular tissue. The splenic pulp was distributed in the whole spleen, which could be divided into white pulp and red pulp. The red and white pulps showed mixture distribution without obvious boundary (Fig. 3). The red pulp accounted for most of the spleen parenchyma, which was mainly composed of dense red blood cells. The splenic sinus was observed, while the splenic cord was not obvious. The white pulp with deeper staining was mainly composed of dense lymphocytes. The nuclei of lymphocytes were stained dark purple. After Gordon-Sweet silver staining to show reticular fibers, splenic nodules could be clearly observed (Fig. 4). Therefore, the red pulp and the white pulp could be clearly distinguished. Splenic nodules were similar with those of mammals, which were mainly composed of dense lymphocytes. Acid phosphatase staining revealed that there were many phagocytic cells in zebrafish spleen (Fig. 4). Electron microscopy showed the red and white pulp ultrastructure of the spleen. There were different types of red blood cells, platelets and plasma cells in the red pulp, and there were lymphocytes, macrophages, monocytes, neutrophils, plasmocytes and reticular cells in the white pulp. The structure of ellipsoid was not observed in the spleen. Multiple vesicles were observed in the lymphocyte cytoplasm, which may be closely related to the immune function of lymphocytes (Fig. 5﹣10). The spleen of zebrafish was similar to that of most bony fish, and the ellipsoid was not observed.

Abstract:As one of the most prominent external morphological features, color patterns play an important role in animal survival and communication. In order to understand morphological changes in the early development of Siniperca chautsi, CCD-Adapter anatomical microscope was used to observe the formation, distribution and main patterns (stripe, band and spot) of the pigment cells in the early stage, from embryo stage up to 40 days after hatching. At the same time, the skin was observed after tissue section. The results show that melanophores are first observed in the yolk sac and oil balls at embryonic stage, xanthophores appear on the head at the pre-hatching stage. After hatching, melanophores develop most prominently and erythrophores appear behind the eye and in the front of trunk (Fig. 1). After 5 days of age, xanthophores increase throughout the whole body and melanophores continue to develop and pattern formation begins (Fig. 2). Early color pattern formation processes of S.chautsi include: 1)Trunk band: some melanin on the back at 5 days old, the dorsal melanin and the abdominal melanin are connected at 14 days old (Fig. 4a﹣c). 2) Eye strip: the melanin above the gill cover increases at 10 days old, the hind strip part forms from the upper edge of gill cover to the front of the back at 12 days old, the front strip part forms from the upper jaw to the front of the eye at 17 days old (Fig. 4d﹣f). 3) Overhead strip: the black spots above the top of the head increase at 6 days old, these spots extend forward and backward at 18 days old. The black strip just above the top of the head forms at 23 days old (Fig. 4g﹣i). 4) Trunk spots: a small black spot appears at the bottom of the tail at 8 days old, three irregular black patches appear in the front of the tail at 15 days old. Five round black spots formed in the back of the torso at 25 days old (Fig. 4j﹣l). The results show that melanophores, xanthophores and erythrophores appear at S.chautsi′s body surface in early stage, the whole body is dominated by black color, the main pigment strips and spots gradually form in different modes at the later larval stages, and pigment layer composition and distribution are different in different skin parts.

Abstract:We aimed to investigate the expression of Piwil2, Stat3 and Bcl-2 proteins and their location correlation in sterile male mice. Methods: 60 male Kunming mice were randomly divided into the experimental group and the control group, 30 in each group. The sterile mice were modeled with administration of Chinese herb Tripterygium wilfordii for 28d. The control group was intragastrically administered with the same dose of normal saline for the same duration. After modeling, two groups of male mice were mated with female mice respectively. The samples are respectively detected for expressions of Piwil2 protein, Stat3 protein and Bcl-2 protein by immunohistochemical staining and Western blot. We compared the expressions of these three proteins between the two groups. The HE staining showed that the structure of seminiferous tubules in the testis tissue of the experimental group was significantly damaged, the number of spermatogenetic cells including spermatogonia and primary spermatocytes was significantly reduced, and the fertilization ability was significantly decreased after mating, indicationg that the model establishment was successful. The results of IHC staining showed that the expressions of Piwil2 protein, Stat3 protein and Bcl-2 protein and the number of positive cells in the experimental group were significantly lower than those in the control group (P < 0.001) (Table 2). Western blot results also showed significantly decreased expressions of these three proteins in the experimental group than in the control group (Piwil2 protein: P < 0.05, Stat3 protein: P < 0.05, Bcl-2 protein: P < 0.01) (Fig. 3, Fig. 4). This study shows that the expressions of Piwil2, Stat3 and Bcl-2 proteins in the testis may play important roles in the occurrence of infertility.

Abstract:Reishia clavigera (Mollusca: Muricidae) is a common species in the rocky intertidal zone along the coastal islands of Wenzhou, Zhejiang Province in China. There are many islands and their hydrological environments are complex in coastal region of Wenzhou. In order to investigate the genetic diversity and population structure of R. clavigera, we collected 158 individuals from 14 populations (Table 1 and Fig. 1). We amplified 672 bp length of mitochondrial cytochrome c oxidase submitⅠ(COⅠ) gene (primer LCOⅠ 1490 5′-GGT CAA CAA ATC ATA AAG ATA TTG G-3′and HCOI2198 5′-TAA ACT TCA GGG TGA CCA AAA AAT CA-3′), which could provide the basic data on their resources and genetic utilization. The ratios of A, T, C, G were 23.0%, 38.4%, 17.4% and 21.2% (Table 2)，respectively, which was in accordance with the mitochondrial composition. All sequences obtained included 108 polymorphism sites (57 parsim-information sites and 51 singleton sites) (Table 2). The nucleotide diversity was 0.008 0, while haplotypes diversity and the average number of nucleotide differences were 0.978 and 5.328, respectively (Table 2). A total of 102 haplotypes were detected (Table 3). The genetic distance between populations ranged from 0.005 7 to 0.011 1, and the genetic distance within the population ranged from 0.005 7 to 0.010 8 (Table 4). The genetic distance between groups from Niyu Island and Dazhushi Island was minimal, and the genetic distance within groups from Dongtou Island was maximal. The correlation between genetic distance and geographical distance based on COⅠ gene fragments was significant (R2 = 0.134, P < 0.01), but there was no geographical isolation (r = 0.365, P = 0.11) (Fig. 2). The topological structure of R. clavigera in ML analysis had no obvious geographical structure (Fig. 3 and Fig. 4).

Abstract:We investigated fecal microflora in the fecal samples of three species of Elaphe (E. carinata, E. schrenckii and E. anomala) which were fed in captivity in laboratory by using a V4﹣V5 region of 16S rRNA gene amplicon sequencing and analyzed the diversity and composition of the microflora. The results showed that the number of OUTs was 192 and the number of shared OTUs was 116 in all samples (Fig. 1). The diversity indexes (Shannon and Simpson) were significantly different between the E. carinata and the E. schrenckii, but the E. anomala had no significant difference when compared to the above two species. There was no significant difference in ace and chao1 among the three species (Table 1). Eight phyla were detected in the samples, but only three phyla, Firmicutes, Bacteroidetes and Proteobacteria were shared by these three species (Fig. 3). The ranking of relative abundance of the three phyla was obviously different. There were 4 genera (Bacteroides, norank_f_Porphyromonadaceae, Enterococcus, Escherichia-Shigella), and the average relative abundance was 77.92% in the E. carinata faeces, there were 9 genera (Bacteroides, Bacillus, Enterococcus, Clostridium_sensu_stricto_1, Clostridium_sensu_stricto_11, Paeniclostridium, Aeromonas, Escherichia-Shigella, Proteus, total of average relative abundance is 75.53%) in the E. schrenckii faeces. There were 6 genera (Bacteroides, norank_f_Porphyromonadaceae, Enterococcus, Clostridium_sensu_stricto_1, Edwardsiella, Escherichia-Shigella), and the average relative abundance was 67.42% in the E. anomala faeces (Table 2). Notably, Escherichia-Shigella had the highest relative abundance in E. carinata faeces; Clostridium_sensu_stricto_11 had the highest relative abundance in E. schrenckii faeces; Bacteroides had the highest relative abundance in E. anomala faeces. There is significant difference in the composition of microbial communities in the faeces among the three species. Under the same feeding environment and food condition, the main factors determining the difference of the microbial communities may be closely related to the specific genomes of each species, which needs further clarification.

Abstract:When Tetrahymena cells are induced to mate for sexual reproduction in the lab, the resulting population always contains a certain proportion of unmated parental cells besides sexual progeny. However, highly purified sexual progeny is an important basis for studying some basic biological questions of Tetrahymena. In this study, based on the characteristics that mating Tetrahymena loses the ability to ingest particles and the newly produced sexual progeny does not recover this ability within several hours, we added the Fe3O4 magnetic nanoparticles to the mating population of T. thermophila at the late stage of sexual reproduction and only the unmated parent cells would become magnetic because they still had the ability to ingest the magnetic particles, which made possible an efficient isolation of sexual progeny under the action of magnetic field. By optimizing the time point at which cell population was passed through the separation column, the diameter of the column and the quality of iron/quartz powder in the column, we could make the purity and yield of the sexual progeny both more than 98%. The established method can also be used for the separation and purification of the sexual progeny of other Tetrahymena or ciliate species.

Abstract:The Saiga tatarica, a member of the Bovidae, is classified as critically endangered A2acr ver 3.1 by the IUCN. It was historically distributed in northwest of China and was extirpated in the mid-twentieth century in wild. In order to initially explore the effects of the partial biological indicators for the newborn S. tatarica, the survival ratio, sex ratio, single and double lamb ratios, weight and others were included. From the end of April to the start of May 2018, this study has conducted on-site measuring of 61 newborn S. tataricas in the Gansu Endangered Animal Protection Center which was the only artificial domesticated base of the S. tatarica in China, and has conducted an inductive analysis of the measurements. The results showed that the period of childbirth began from April 25 to May 5, 2018, a total of 61 lambs were delivered during the period. The peak period of lambing production was concentrated from April 28 to May 2 (Fig. 1). The newborn S. tatarica survived 56, with the survival ratio of 91.80%. The newborn male lambs accounted for 40.98% of the total lambs, and the newborn female lambs accounted for 59.02% of the total lambs, and the female-male sex ratio (♀︰♂) was 1.44︰1 (Table 2). The differences were not significant after χ2 test (P > 0.05). The single lamb ratio was 80.33%, the double lamb ratio was 19.67%. The differences were extremely significant between the ratio of single lamb and double lamb (P < 0.01). The average weight of newborn lambs were mainly concentrated in 2.501﹣3.000 kg, and the average newborn weight of male lambs was slightly higher than those of female lambs, the average newborn weight of single lambs was slightly higher than those of double lambs. The differences were not significant that the average newborn weight of the single lamb and the double lamb of the newborn male S. tataricas. The differences were not significant that the single lamb and the double lamb of the newborn female S. tataricas, and the differences were not significant that the total average newborn weight of the single and double lambs and the total average weight of male and female lambs (P > 0.05, Table 3). Those research results provided a reference for the basic research of S. tatarica in China and provided a theoretical basis for S. tatarica population of protection.

Abstract:The crocodile lizard (Shinisaurus crocodilurus) mainly preys on small animals including insects, spiders, earthworms, frogs and fish, but whether it can prey on relatively large animals like other reptiles is unknown yet. Here we reported for the first time that crocodile lizards preyed on an oriental garden lizard (Calotes versicolor) and a greater green snake (Cyclophiops major). The predation process was observed and analyzed using all occurrence sampling. This discovery indicates that the crocodile lizard has a wide diet, and is capable of preying on large animals like lizards. Therefore, we recommend that a diversity of foods should be provided to crocodile lizards in captivity to avoid malnutrition.

Abstract:Vocal communication and seasonal breeding are typical characteristics of frogs. During the breeding season, males compete for territories and attract females by advertisement calls and females evaluate the individual of males by their characteristics and resources possession and mate choice was expressed by phonotactic behavior. Parameters of the phonotactic behavior were usually used to test the choice preference and behavioral response of the acoustic signal. Auditory and motor pathways are the basis of the neural mechanisms of the phonotactic behavior in frog (Fig. 1, 2). In the present study, we reviewed the related researches on the phonotactic behavior of frogs, and made a summarization on the mechanism, evaluation methods as well as the possible factors that may influence on the phonotactic behavior. At the end of this paper, we provided some suggestions for the future research on the phonotactic behavior.

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