Abstract:Foraging is a fundamental behavior to ensure animals’ survival and reproduction. Foraging decisions should balance the risks of starvation and predation which usually varied both spatially and temporally in the wild. The energetic state-risk allocation hypothesis presented that the optimal trade-off between energetic gain from foraging and avoidance of predation should depend on animals’ current physiological states (e.g., hunger), on current environmental states (e.g. actual predation risk, food availability), and perhaps also on past and expected future states. However, in terms of the model of energetic state and predation risk allocation, studies in rodents have yielded mixed results. Therefore, we used a Y-shaped maze by a 10-min behavioral test to measure the discrimination to the predator odor of a Steppe Polecat (Mustela eversmannii) or no-predator (i.e. horse) odor in female Mongolian Gerbils (Meriones unguiculatus) deprived food for 18 h in field laboratory conditions. Then we investigated the feeding and antipredator efforts of the fasted female gerbils in the presence of predator odor with food or those of in the control no-predator odor with food at a neutral arena. We aimed to test the hypothesis that acute predation risk inhibited the foraging effort of the starving gerbils, and then to address the trade-off between the starvation and predation risk in the foraging decision of gerbils, as a desert rodent. We found that both the frequency and the duration of visiting the predator odor selection box was significantly less than that of visiting the control box (Wilcoxon signed ranks test: frequency, Z = 2.405, P = 0.016; duration, Z = 2.803, P = 0.005; Table 1), which indicated a remarkable aversive or fear response to the steppe polecat odors in female Mongolian gerbils. Furthermore, we observed that starving gerbils displayed their foraging effort both in the treatment with a predation risk and without predation risk, e.g. unchanged their foraging frequency compared to under the predation risk and under the control situation (Mann-Whitney test: Z = 1.514, P = 0.130; Fig. 2). It was just that，compared with the foraging latency (58.6 ± 35.5 s) under control situation，the starving gerbils under predation risks increased the foraging latency (110.4 ± 57.9 s), delaying their foraging (Mann-Whitney test: Z = 2.068, P = 0.039; Fig. 1), yet decreased the duration per foraging (Mann-Whitney test: Z = 2.856, P = 0.004; Fig. 1). Additionally, we noted that gerbils were higher in the effort of the up-investigating to the space for vigilance (Mann-Whitney test: frequency: Z = 3.187, P = 0.001; Fig.2; duration, Z = 2.725, P = 0.006; Fig. 3) and self-grooming for releasing fear in the treatment with the predation odor risk (Mann-Whitney test: Z = 2.289, P = 0.022, Fig. 2). Our results, thus, revealed that the acute predation risk could not completely inhibit the foraging activities in starving female Mongolian gerbils, and partly supported the energetic state-predation risk allocation hypothesis. These results suggested that the gerbil could respond to the hazard of starvation and predation by risk-adjusting or balancing, and trade off the energetic gain from foraging and cost of predation maximizing the current or lifetime fitness, which benefited to the Mongolian gerbils adapt to desert environment characterized by largely unpredictable food resources and relative higher predation-risk.

Abstract:Automatic acoustic recorder is a low-cost, high-efficiency research equipment that can effectively monitor the activity level of species in a large space range and long-time span. To understand the rhythm of the Galliformes birds, vocalization of Brown Eared Pheasant (Crossoptilon mantchuricum), Koklass Pheasant (Pucrasia macrolopha) and Ring-necked Pheasant (Phasianus colchicus) were recorded using 40 automatic recorders in the Xiaolongmen National Forestry Park during 2019 breeding season (from April 27th to June 31st). The calls of Brown Eared Pheasant, Koklass Pheasant and Ring-necked Pheasant were extracted based on human-assisted machine learning. By optimizing the time-frequency parameters of the sound extraction, the correct rates of call extraction of the Koklass Pheasant and Ring-necked Pheasant were 73.32% and 89.32%, with the detection rates of 52.91% and 67.36%; however, the correct rate of call extraction of Brown Eared Pheasant was only 8.69% with the detection rate of 58.54%. Peak frequency, syllable duration, number of syllables and strophe duration were measured for each species. Acoustic characteristics were compared among species using K-S test and F-test. The calls of Brown Eared Pheasant were occurred rich in harmonics, while Koklass Pheasant and Ring-necked Pheasant were relatively few harmonics (Fig. 1, Table 1). Both breeding rhythms and daily rhythms on vocal activity were compared among species using T-test. In terms of breeding rhythms, all these three pheasant species had a peak of vocal activity during mating and early hatching periods (Fig. 2); in terms of daily rhythm, the Brown Eared Pheasant and the Ring-necked Pheasant had two peaks in the morning and evening, while Koklass Pheasant only had one peak of dawn calling (Fig. 3).

Abstract:In this study, the daily vocal pattern of female Common Cuckoos (Cuculus canorus) were described. Recordings were made using 28 passive acoustic recorders during breeding season in Wild Duck Lake Wetland Park, Beijing (10 recorders, from 7th May to 8th July in 2017), Liaohe Delta Nature Reserve, Liaoning (10 recorders, from 28th June to 29th July in 2018), and Dagangzi Forest Farm, Jilin (8 recorders, from 17th May to 10th July). A total of 2 777 female calls (Fig. 1) were detected, with 1 431 calls from Wild Duck Lake Wetland Park, 1 222 calls from Liaohe Delta Nature Reserve, and 124 calls from Dagangzi Forest Farm. The females in Wild Duck Lake Wetland Park and Liaohe Delta Nature Reserve showed a similar pattern of daily vocal activity, reaching a sharp peak in the dawn (Fig. 2a, b); while females in Dagangzi Forest Farm showed two peaks of daily vocal activity, with peak activity occurring around early morning and dusk (Fig. 2c). Through comparing these daily vocal patterns with daily egg-laying pattern (Fig. 2d), we suggest that the function of female call in the study populations is to advertise territory and attract mates, rather than misdirect host defenses. This study helps to understand the function of the female Common Cuckoo call.

Abstract:The accumulation of large amounts of dung from captive herbivorous wildlife can easily cause environmental pollution and its treatment is time-consuming. Dung beetles feed on the dung of mammals or use mammalian dung as a place for breeding activities If dung beetles can be screened and used to treat the dung of herbivorous wildlife, it will undoubtedly have certain application value. Fresh dung samples of three species of captive herbivorous wild mammals (namely, Elephas maximus, Dama dama and Giraffa camelopardalis) from Nanjing Hongshan Forest Zoo were used as bait in this study from August, 2019 to September, 2019. The replacement and placement trap methods during the two stages of the experiment were used in Nanjing Zijin mountain to investigate the attraction of dung to dung beetles. One-way ANOVA was used to test the trapping quantity of dung beetles. The results showed that a total of 4 597 dung beetles were recorded in the first stage, belonging to 7 species, 5 genera and 1 family. During the first stage, Onthophagus sinicus (48.79%) and Caccobius brevis (30.95%) were the dominant species (Table 1). In the second stage, 3 512 dung beetles were recorded, belonging to 7 species, 5 genera and 1 family (Fig. 2). Dung beetles showed polyphagous characteristics to the dung of three species of wild mammals and the effective time periods of feeding activities in all samples of dung were mainly concentrated in 1﹣2 d, which occupied a short time. There was almost no dung beetles found in the dung on the 7th day (Fig. 1, 3). Diversity index pattern of dung beetle community shows that Shannon-Wiener index and Pielou index were relatively higher in E. maximus dung (Fig. 4). The study shows that the dung of three species of herbivorous wild mammals in captivity from zoo have great attraction to dung beetles and further study is needed on the treatment effect on dung by dung beetles.

Abstract:To promote the heath assessment and medical care of captive Bornean Orangutans (Pongo pygmaeus) and provide basic parameters for supplementing the hematological database, a total of 24 blood samples from 18 healthy orangutans were collected from 6 zoos and routine and biochemistry parameters of these samples were analyzed. The subjects were divided into different groups based on sex (14 male, 10 female) and age (7 infants, 6 subadults, 11 adults), and blood parameters of different groups were compared. The results showed that the blood routine parameters of male and female orangutans were not significantly different with each other (Table 2). The lymphocyte (LYM) density decreased with age (One Way ANOVA test, F = 5.200, P < 0.05) (Table 3). Male individuals showed higher alanine aminotransferase (ALT) density than that of female (One Way ANOVA test，F = 4.911, P < 0.05), while other parameters showed no sexual difference (One Way ANOVA test, P > 0.05) (Table 4). Total protein (TP) showed significant differences among age groups (One Way ANOVA test, F = 4.715, P < 0.05) (Table 5). There was a high glucose (GLU) content in the blood of orangutans in Chinese zoos, which should be paid attention in daily husbandry.

Abstract:The utilization of various agricultural pesticides has both direct and indirect effects on aquatic animals. The toxic effects of organochlorine insecticide endosulfan on tadpoles of the Chinese tiger frog (Hoplobatrachus chinensis) were investigated in this study. Specifically, we first examined the safe concentration (SC) and evaluated the toxic rank of endosulfan in an acute toxicity assay. We then examined the blood biomarker (erythrocyte nuclear abnormality) and determined the metabolic enzyme activitie of acid phosphatase (ACP), alkaline phosphatase (AKP) and lactate dehydrogenase (LDH) in a chronic toxicity test. In terms of the acute toxicity, the average mortality percentage of tadpoles significantly increased with increasing concentrations and exposure duration of endosulfan solution, indicating dose- and time-dependent lethality of this drug (Table 1). The half lethal concentration (LC50 value) of endosulfan for H. chinensis tadpoles at 96 h was 23.38 μg/L and the safe concentration was 2.34 μg/L (Table 2). Five different types of erythrocyte nuclear abnormalities were observed: broken nucleus, binucleated, unequal division, karyopyknosis and anucleated (Fig. 1). The total frequencies of abnormal erythrocytes were positively correlated with pesticide concentrations (Table 3). Activities of all three enzymes were influenced. Compared with the control treatment, the activities of both ACP and AKP were decreased with increased endosulfan concentrations, while LDH activity was first increased and then decreased (Table 4). These findings suggest that endosulfan has a high toxicity for H. chinensis tadpoles and that the erythrocyte nuclear abnormality and particular metabolic enzymes may be considered as biomarkers for environmental monitoring.

Abstract:Hynobius maoershanens belongs to Hynobiidae, and is endemic to Maoershan Natural Reserve in Guangxi Province, China. It is restrictedly inhabited in alpine swamp at an altitude of 2,000 m with a small population size. We investigated the histological structure of liver, using anatomical methods and paraffin sectioning technique. The total length, head length, head width, tail length, body weight and liver weight were measured. An Olympus BX-63 automatic fluorescence microscope was used to analyze the liver histological structure. The results showed that the histological structure of the liver in H. maoershanensis was similar to that of other amphibians, which could be divided into the right and left lobe (Fig. 1a). Glisson capsule, central veins, portal areas and hepatic cells constituted the liver structure (Fig. 2a, b). The hepatocyte cords were arranged in an indistinct radial pattern around the central vein (Fig. 2c, d). Moreover, interlobular veins, bile ducts in portal areas were clearly found (Fig. 2e). The boundaries of liver lobules were unclear due to the underdevelopment of connective tissues (Fig. 2c-g). Many brown-black melanin granules could be seen in liver parenchyma (Fig. 2a, b). Our findings suggest that the brown-black melanin granules in the liver parenchyma of H. maoershanensis might be an adaptation to the low-oxygen environment.

Abstract:To study effect of dietary Inulin on the growth, digestive function, and immunity of juvenile Takifugu obscures, a total of 640 juvenile T. obscures (initial body weight of 6.97 ± 1.32 g) were randomly divided into 4 groups with 4 replicates and data were analyzed by single factor test. Four groups were fed diets with 0% (control group), 0.25%, 0.5% and 1% inulin, respectively. After rearing 8 weeks, the growth, nutritive composition, activities of digestive enzymes, nonspecific immunity enzyme were investigated. The supplementation of Inulin had no significant effect on growth index of juvenile T. obscures (P > 0.05). The crude fat content in the white muscle of 0.25% group (0.87%) was significantly higher than that of 0.5% group (0.82%) and 1% group (0.76%), and the crude protein content in the white muscle of 0.25% group (18.75%) was significantly higher than that of 1% group (18.50%), but showed no significant difference compared with the control group (18.62%) and 0.5% group (18.60%); The activities of pepsin, trypsin, lipase and amylase in intestine were increased with the increase of inulin addition. The activities of glutathione peroxidase and superoxide dismutase in the liver of 0.5% group were significantly higher than in the rest three groups, and the difference was very significant (P < 0.01). The activity of catalase in intestine of 0.5% group was very significantly higher (P < 0.01) than that of 0.25% or 1% group, while the malondialdehyde content was decreased with the increase of inulin addition. Therefore, the added amount of inulin should be 0.5%, a concentration that that cannot promote the growth significantly, but improve the activities of digestive enzymes and nonspecific immunity enzymes of juvenile T. obscures.

Abstract:In order to study the function V-ATPase H gene expression on salinity stress in Sinonovacula constricta, the adults were used as experimental materials, and the stress was tested in different salinities (5, 15, 20, 25, 35). The freezing point osmometer was used to determine the difference in serum osmotic pressure, and V-ATPase ELISA detection kit was used to determine the activity of V-ATPase. The full-length ORF sequence of V-ATPase H gene was closed and its mRNA expression was analyzed. The results showed that the serum osmolarity of the low-salt group (salinity 5, salinity 15) and that of high-salt group (salinity 25, salinity 35) changed significantly, both were very significantly different from the control group (salinity 20) (P < 0.01). With the treatment time, the V-ATPase activity of the experimental group showed a decreasing first and then increasing trend, while the control group showed no significant change. The open reading frame (ORF) of the V-ATPase H gene was 1 440 bp in length, encoding 479 amino acids. The qPCR results showed that the expression level of V-ATPase H gene in the gills of S. constricta was significantly higher than that in the other 6 tissues including siphon, mantle, kidney, digestive gland, flabs, and foot (P < 0.01). Under salinity stress, the expression of V-ATPase H gene in the gills of each experimental group continued to rise, peaking at 24 h, which was significantly higher than that of the control group (P < 0.05). The experimental results show that the V-ATPase H gene plays a role in maintaining the balance between its own serum osmotic pressure and the external osmotic pressure in low-salt and high-salt environments during salinity adaptation of S. constricta.

Abstract:Red Panda Ailurus fulgens which depends heavily on bamboo with high fiber content, like Giant Panda Ailuropoda melanoleuca, retains the similar digestive system with carnivore. The purpose of our research is to determine the microbial structure of the feces of Red panda, and explore the biological mechanism of the bamboo feeding habits. Feces of captive Red pandas from Hangzhou zoo and Shanghai zoo were collected, and Hiseq high-throughput sequencing based on 16S rRNA gene was conducted to determine the structure of fecal microbiota of Red panda. For the diversity of microbiota composition, Sobs, Shannon, simpson, ace and Chao1 index were calculated. Wilcoxon rank-sum test was used to evaluate the influence of diet composition and age on the microbiota structure. Tax4Fun was used to predict the function of microbiota. The results showed that the diet compositions influenced the microbiota diversity of Red pandas (P < 0.05), and the microbial diversity of adult Red pandas was significantly higher than that of cub and old groups (P < 0.05) (Table 2). The fecal microbiota of the Red panda was mainly composed of Firmicutes, Proteobacteria and Bacteroides at phylum level, and composed of Eschericia-Shigella, Clostridium sensu stricto and Enterobacter at genus level (Fig. 2). For the fecal microbiota of Red panda, the metabolic genes related to carbohydrates and amino acids were the richest, followed by the metabolic genes related to energy and lipid (Fig. 5). 6-phosphate-β-glucosidase and β-glucosidase might be helpful for the digestion of high fiber diets (Fig. 6). The microorganisms of Red panda can be helpful in metabolizing carbohydrates and amino acids, and producing enzymes to digest cellulose.

Abstract:Environmental DNA (eDNA) detection method is considered as a new survey tool for the wildlife aquatic animals in recent years. To further investigate the Hong Kong newt (Paramesotriton hongkongensis) population distribution in the wild, we thus aimed to design a set of specific primers and TaqMan probe and verify the feasibility of this eDNA method for water samples. The mitochondrial cytochrome b genes of 11 Paramesotriton species were sequenced, and then the primers and TaqMan probe were designed using Primer Express Software 3.0. The primers and TaqMan probe specificity were verified by comparing to the NCBI GenBank and testing with the annealing temperature gradient. The qPCR amplification efficiency was optimized by testing different concentrations of primers and TaqMan probe. Then, eDNA was detected in the water samples in aquarium tanks with different numbers of Hong Kong newt to assess the sensitivity of the established qPCR method. Also, the decay rate of Hong Kong newt eDNA was determined to estimate its lasting time in water. The results showed that the designed primers and TaqMan probe only amplified eDNA of Hong Kong newt but not the other 10 closely related newts (Table 3 and 4). The optimized qPCR achieved the efficiency of 93.9 % and the limit of detection down to 10 DNA copies (Fig. 2 and Table 5). The established qPCR method was sensitive enough to detect 13.56 ± 3.35 DNA copies/ml in the aquarium tank with 1 Hong Kong newt after housing for 24 h (Fig. 3). In addition, the decay rate experiment demonstrated that the use of 0.45 μm pore size filter membrane was effective to detect and monitor the Hong Kong newt eDNA in 15 d (Fig. 6). In this study, an eDNA detection method was successfully designed and established to detect the presence or absence of Hong Kong newt in water environments, which can be potentially utilized in the field inspection.

Abstract:In order to study the expression of PKCθ and Spartin in Nile tilapia (Oreochromis niloticus), these two proteins were expressed in Escherichia coli, purified, and then used to immunize Japanese big-ear white rabbits (Oryctolagus cuniculus) according to the conventional method to prepare rabbit anti-PKCθ and rabbit anti-Spartin polyclonal antibody. The titers of rabbit anti-PKCθ and rabbit anti-Spartin antiserum were evaluated by indirect ELISA. The expressions of these two proteins in different issues (liver, spleen, intestine and muscle) of Nile tilapia were detected by Western Blotting. The results showed that the prokaryotic expression vectors of pET-B2m-PKCθ and pET-B2m-Spartin were successfully constructed, the recombinant PKCθ and Spartin proteins were expressed in inclusion body with molecular weight of 56 ku and 30 ku, respectively. The indirect ELISA assay showed that the rabbit anti-PKCθ and rabbit anti-Spartin antisera had a good sensitivity with the titer of 1︰512 000, and Western blotting showed that the polyclonal antibody had good specificity. PKCθ protein was expressed in the liver, spleen, intestine and muscle. Spartin protein was only expressed in the muscle. In this study, the PKCθ and Spartin recombinant proteins of Nile tilapia were expressed and purified successfully, the polyclonal antibodies against PKCθ and Spartin proteins with a high titer were obtained, and the expressions of PKCθ and Spartin in different issues of Nile Tilapia were determined. The current study will shed new light on the functional and mechanism study on PKCθ and Spartin in Nile tilapia.

Abstract:The flight ability, nocturnal activity pattern and similar morphological characteristics of Chiroptera (bats) increase the difficulty for researchers to accurately identify them in the field. Therefore, extensive investigation should be performed to identify the diversity and accurately distribution information of Chiroptera. In August 2019, we sampled six Horseshoe Bat (Rhinolophus spp.) in Chaoyang of Liaoning province in China (119°08′21″ E, 40°53′15″ N). Based on morphological data (Table 1), barcode sequence (COI) (the accession number in NCBI database is MT240893) and phylogenetic relationship (Fig. 1), a Horseshoe Bat was identified as a Big-eared Horseshoe Bat (R. macrotis). This is the first record of Big-eared Horseshoe Bat in Liaoning, and the northernmost record of this species in China.

Abstract:Specimen resources are becoming more and more precious, so specimen repair techniques need to be constantly updated. This paper addresses the phenomenon of dry cracks, hair removal, discoloration, mildew, and worm-eaten of peeled specimens using improved techniques, which includes use of new materials, way of addition color, mildew resistance and mothproof. Compared with the previous techniques, the new techniques fully utilizes the new materials and ways, which are simple and practicable. Several cases of specimen repairing were introduced here, results show that the new techniques can restore damaged specimens as before.

Abstract:Based on The Mammalian Fauna of Guizhou, historical accounts and the latest research on the taxonomy and phylogeny of animals, we provided an updated species checklist with the references of mammal species in Guizhou Province, China. As of December 31, 2019, the new mammalian inventory of Guizhou has 9 orders, 29 families, 84 genera and 153 species, including 56 of bats, 38 of rodents，24 of carnivores, 19 of insectivores, seven of ungulates, four of primates, three of hares, one of scandentia and pangolins. When compared to mammal species the records in The Mammalian Fauna of Guizhou, we added 35 species, including 5 new species: Murina shuipuensis, M. fanjingshanensis, M. rongjiangensis, M. liboensis, Apodemus nigrus, one new species records in country (M. eleryi), 14 new record species from Guizhou, and 17 species were confirmed to be distributed in Guizhou. At the same time, 23 species of animals should be removed from the Guizhou animal List due to errors in specimen identification，synonyms，or confirmed not distributed in Guizhou. 19 species are listed at the National Key Protection Species Lists, including 5 species I level and 14 species as II level. According to the IUCN Red List of Threatened Species, one mammal species is Critically endangered (CR), 5 mammal species are Endangered (EN), 5 mammal species are Vulnerable, 9 mammal species are Near Threatened. On the basis of the Red List of China’s Vertebrates, 6 mammal species are Critically endangered, 5 mammal species are Endangered (EN), 17 mammal species are Vulnerable, 39 mammal species are Near Threatened.

Abstract:The male sex differentiation in crustaceans is mainly regulated by the insulin-like androgenic gland hormone (IAG) secreted by the androgenic gland (AG). In Macrobrachium rosenbergii, manipulation of the IAG can lead to a full-functional reversed female, which is used to produce all-male offspring. Therefore, the IAG-based sex regulation technique has its application potential. At present, IAG has been reported in many economic crustaceans. It is found that IAG expression is not limited to the AG, and its function is more extensive. With the application of RNA interference technology in aquatic animals, the study of gene function is more easily realized. Therefore, the signaling mechanism of IAG function and its upstream regulatory network have gradually become a hot topic. This paper reviews the recent research progresses in the regulation of gene expression, molecular characteristics and biological functions of IAG, as well as the signaling mechanisms of IAG function. Summary of these researches will provide a basis for further elucidation of the physiological function of IAG, as well as its mode of action.

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