Abstract:Stopover sites are essential for successful migration, as migrating birds rely on these sites to refuel and rest. In order to get the stopover sites information of Common Cranes (Grus grus), we tracked 4 birds at Caohai National Nature Reserve using satellite tracking devices in winter of 2013 and 2015. Our results showed that 3 of them stayed in the Yellow River (37°53′﹣37°75′N, 105°74′﹣105°94′E) and Qingshui River (37°11′﹣37°35′N, 105°64′﹣105°82′E) wetlands in Zhongwei City, Ningxia, and this area would be important for the migration of Common Cranes (Fig. 1). During spring stopovers, the cranes arrived there around early February to early March and stayed for 24.5 ± 6.7 d on average (Range: 15﹣30 d, n = 4) and left around late February to late March. During autumn migration, cranes stayed there for 32.0 ± 4.2 d on average (Range: 29﹣35 d, n = 2) from late October or early November to the late November (Table 2). During their stopover, the cranes were mainly located in Changshantou Reservoir, eyots of Yellow River in Zhongning County, and areas along the Qingshui River in Tongxin County (Fig. 2﹣4). We observed 636 cranes at the night roosting site near Changshantou reservoir on 18 March in 2017 (Table 3). These stopover sites were still under no protection or management. We suggest local forestry department supply supplementary food (e.g., corn) during the migration period to reduce the mortality of cranes for feeding corn seeds coated with pesticide.

Abstract:To understand the behavior patterns, including the time budget of endangered animals is important to make better conservation measures. During March to October in 2016 and 2017, we observed the behaviors of 4 pairs of Cinereous Vulture (Aegypius monachus) and their nestlings during the nestling period, using the focal sampling method and all occurrence recording method, in Hejing County, Xinjiang, China. The study area is located in the Tianshan Mts (42°50′﹣43°02′N, 86°03′﹣86°32′E). Two or three infrared cameras were set up around each nest and about 110 thousand photos with over 90 h videos were captured. We also observed the behavior of parents and nestlings, 400﹣600 m far away from the nest, for 35 d (412 h) during the nestling period. All the behaviors of Cinereous Vulture, defined by the postures and actions, were classified into 9 categories (including 33 behaviors) for parents and 6 categories (including 28 behaviors) for nestlings according to the motivations and functions. During the nestling period, caring, alerting and looking of the time budget of parents and resting and looking of nestlings were dominant. The times of feeding the nestlings from the parents reached the peak at 12:30﹣13:30, followed by the second peak at 15:30﹣18:30 (Fig. 2). The length of nestling period of Cinereous Vulture lasted for (97 ± 6.16) d (90﹣105 d), and we divided the entire nestling period into three stages (Pre-nestling: April and May, Mid-nestling: June, Post-nestling: July) and analyzed the time budget of parents and nestlings during the three stages (Table 1 and 2). The time budget of pre-nestling period was analyzed according to the videos, and the other two stages were analyzed according to the observation supplemented with the data of cameras by time. The differences of these budgets were tested by one-way ANOVA. We found that: there was no significant difference between the time budget of pre-nestling and post-nestling of parents (P > 0.05) while either the difference between pre-nestling and mid-nestling or between mid-nestling and post-nestling was significant (P < 0.01); the difference between any two stages of nestlings was significant (P < 0.01). During the post-nestling period, the defecating numbers of nestling (3.52 ± 1.92) was much more than the feeding numbers from parents (0.93 ± 0.96) which, we believed, was related to the behavior of food storage and bad weather (Fig. 3). As the nestling period progressed, the time of first return of parents with food was gradually delayed (Fig. 4), we suggested that lack of food and anthropogenic factors might influence the breed success of the Cinereous Vulture in Xinjiang.

Abstract:Time budget and behavioral rhythm of animals can be regarded as a kind of behavioral adaptation to environmental conditions. The Black-necked Crane (Grus nigricollis) use fixed roosting sites during overwintering periods and have a daily behavioral pattern of flying out from the roosting sites in the morning to forage and flying back in the evening to roost. To explore the time budget and factors influencing this behavior during different periods of winter, a field study by means of instantaneous scan sampling was conducted on the flight and behavior patterns of Black-necked Cranes at seven roosting sites at the Caohai wetland. The field observations were conducted in the whole winter, which divided into three periods: early winter (Nov. 9﹣Dec. 31), mid-winter (Jan. 1﹣Feb. 21) and late winter (Feb. 22﹣Mar. 31). Based on the known behavioral spectrum of Black-necked Cranes and previous observation results (Li et al. 2005), crane behavior at roosting sites before the morning departure and after the evening return was classified into 8 categories and 14 types (Table 1). One-way analysis of variance (ANOVA) was used to test the time differences of the daily departure and return flights between three periods of winter. The results showed that both departure and return times were significantly different between the three periods. Compared with early winter, the departure time from roosting sites was delayed in middle winter and advanced in late winter (mean times: 7:34, 7:40, and 7:13 in the morning), while the return time to roosting sites became gradually later throughout the winter (from 17:12 to 18:15 in the late afternoon) (Fig. 2). A Chi-square R × C table test was used to compare patterns of the departure and return times from roosting sites as well as the behavioral differences before departure and after return between different periods of winter. There was a significant difference of the behavior before departure among early, mid, and late winter (F = 1 768.25, df = 12, P < 0.01), so as to the behavioral differences after their return to the roosting sites (F = 793.98, df = 12, P < 0.01). The behavior of the cranes before the morning departure and after the evening return was significantly different in the early winter period (F = 2 723.16, df = 6, P < 0.01), mid-winter period (F = 1 979.48, df = 6, P < 0.01), and late winter period (F = 5 098.18, df = 6, P < 0.01) (Table 2). The occurrence frequency of various types of behavior in the crane roosting population within 80 min before morning departure and 90 min after evening return were recorded. For the 80-minute period before the morning departure from the roosting site, maintaining (34.32%) and resting (32.38%) were the dominant behaviors, while foraging (43.04%) and resting (23.68%) were the dominant behaviors within the 90-minute period after the evening return (Fig. 3). The Pearson correlation coefficient was used to test the correlation between the flight times and the sunrise and sunset times. The departure times were significantly related with sunrise time (r = 0.832, n = 48, P < 0.01), while the return times were weakly correlated with sunset time (r = 0.353, n = 47, P < 0.01). Multiple linear regression analysis was used to test the effects of temperature and humidity on the departure and return time changes. The difference between the sunrise time and the crane departure time (Y1) was affected by humidity at the time of the departure (W) (Y1 = 0.469﹣0.625W, P < 0.05). The difference between the sunrise time and the crane departure time (Y1) was inversely proportional to humidity at the time of departure (W). The difference between the sunset time and the crane return time (Y2) was affected by mean daily temperature (T) (Y2 = 1.231﹣0.107T, P < 0.05). Our results can be meaningful and useful for further exploring the roosting behavior of Black-necked Cranes as well as their behavioral adaptations to human disturbances.

Abstract:Environment factors affect many larval amphibian traits, including larval growth, development, and time to metamorphosis. Temperature is found to be a key factor in the growth larval among all the environment factors. To uncover the effect of temperature on tadpole growth of Microhyla fissipes, adults M. fissipes collected from Shuangliu, Chengdu, China (30°34′57″N, 103°50′38″E) were used in this work and induced breeding with Luteinizing Hormone Releasing Hormone analogue hormones (LHRHa, dosage: 0.3 μg/g body weight) in laboratory. Forty-five full-sib family tadpoles of stage 28 (stages according to Wang et al. 2017) were randomly and averagely divided into three groups and raised at water temperature (WT) 25 ℃, 28 ℃ and 30 ℃, respectively. Five morphological measurements (Fig. 1) including total length (TOL), body width (BW), interocular space (IOS), snout length (SL) and snout-vent length (SVL) were measured by a stereo microscope with Mshot Image Analysis system (Mc50-N). We used one-way ANOVA and DUNCAN test to analyze the data by the sigmaplot 13.0 software. The growth rate β (%) = (the ending data-beginning data) / beginning data × 100%. Descriptive statistics was presented as mean ± standard error, and the significance level was set at P < 0.05. The results showed that the five measured morphological characters had similar growth trend, of which TOL could be the dominant character to indicate the growth of tadpoles (Fig. 2). The TOL indicated that tadpoles under 25 ℃ grew slowest but presented quickening growth trend after 21 d; tadpoles under 30 ℃ grew fastest but presented slower growth trend after 12 d, and 5 tadpoles were died before end of feeding 28 d; tadpoles under 28 ℃ grew continuously fast and the TOL is longer than those under 30 ℃ at 28 d, also presented quickening growth trend after 21 d, which higher than that under 25 ℃ (Table 1). So, it could be concluded that the survival ratio of tadpoles under 30 ℃ decreased, tadpoles grow slower under 25 ℃, and 28 ℃ is the best feeding temperature for tadpoles of M. fissipes.

Abstract:In order to study reproductive characteristics of Hypophthalmichthys molitrix, 433 samples were collected at the Yichang section in the Yangtze River during March 2014 to September 2016. The data for standard length, body weight, net body weight and gonad weight of each specimen, and eggs′ diameter of 10 females in Ⅳ stage were measured. Egg diameters were measured with the Image-Pro Plus 6.0 software. The gonads′ stage was divided into six stages according to the method of YIN Ming-Cheng (1995). Statistic analysis of all data were taken by Microsoft excel 2007 and SPSS 16.0. The results proved that females spawned from May to August, peaked in June and July. The age of reproductive group consisted of 3, 4, 5, 6 and 7 years, while those at 3 years were consisted a highest percentage. The sex-ratio of females to males was 1.43︰1 (χ2, P < 0.01). A significant differences were found between males and females by the body length to body weight equation (0.01 < P < 0.05, Fig. 2). The Logistic equation was used to calculate the characteristics of the first maturity and found out the minimum size of mature female was 482.3 mm in length and 2 206.7 g in weight, while that of mature male was 484.0 mm in length and 1 677.5 g in weight (Fig. 3). The average egg-diameter in ovary at stage Ⅳ was from 0.57 to 1.332 mm (average 1.01 ± 0.12 mm) and the distribution of eggs diameter showed a one peak curve (Fig. 6). H. molitrix belonged to total spawners based on the gonadosomatic index curve (Fig. 5) and the distribution of the diameter of mature eggs. The absolute fecundity ranged from 102 351 to 767 200 eggs (average 477 662 ± 9 631 eggs) and the relative fecundity per gram ranged from 44.89 to 139.75 eggs (average 93.38 ± 5.92 eggs). The absolute fecundity was power correlated to body length and body weight (Fig. 4). Contrary to the reproductive population of fish in other section of Yangtze River, the average egg-diameter of H. molitrix was relatively smaller, but the fecundity was relatively higher.

Abstract:According to the existing mitochondrial gene sequence of Acrossocheilus parallens, primers were designed, and by using the method of primer walking and PCR amplification, mitochondrial genome sequence of A. kreyenbergii was obtained, and its structure was analyzed. The results showed that the complete mitochondrial genome of A. kreyenbergii was a circular genome, 16 596 bp in length, including 13 protein coding genes, 22 tRNA genes, 2 rRNA genes and two non-coding regions (D-loop and origin of light strand replication, OL). The base composition showed two features, the negative bias for G and AT bias. All protein-coding genes started with ATG as an initiation codon except COⅠ, which used GTG. Two complete termination codons were TAA (38.46%) and TAG (7.69%), and two incomplete stop codons were TA (15.38%) and T (38.64%). A control region of 941 bp in length between tRNAPro and tRNAPhe, a short tandem repeat TA, was found in the 811﹣837 bp intervals. From the information contained in the protein coding gene, the mutation site and the mutation rate of view, ND5, ND4, COⅠ and ND2 were the most suitable molecular markers for interspecific phylogenetic analysis of Acrossocheilus. The phylogenetic tree constructed by Bayesian method for 13 protein coding genes showed that 23 species of Acrossocheilus and Onychostoma were not clustered into monophyletic groups, and could not be clearly distinguished from each other.

Abstract:Erythropoietin (EPO) is a glycoprotein hormone that plays a potential role in hypoxic adaptation in animals. In the present study, the encoding sequences of Epo and Epor genes were cloned in Gymncypris eckloni by RT-PCR method. The coding region of Epo and Epor were 552 bp and 1 590 bp, which encoded 183 and 529 amino acid residues respectively (Fig. 1 and 2). The high homology of EPO and EPOR protein of G. eckloni with other cyprinid fishes indicated that the two proteins were evolutionary conserved (Table 2). Molecular evolution analysis suggested that Epo and Epor genes of G. eckloni did not undergo positive selection (Table 3). Under severe (the dissolved oxygen was 0.3 ± 0.1 mg/L) and moderate hypoxic (the dissolved oxygen was 3.0 ± 0.1 mg/L) conditions, the Epo mRNA levels in key tissues such as brain and red muscle were increased significantly (Fig. 4 and 5), implying its potential roles in movement, neuroprotection and neurotrophy, which may be a regulatory mechanism of fish to hypoxic adaptation.

Abstract:In order to identify a decayed large body of Baleen whale, found at the converging area of Yangtze Estuary and Hangzhou Bay, three regions of mtDNA (control region, cytochrome c oxidase subunitⅠ, COⅠ and cytochrome b, Cyt b) were sequenced. The sequencing results showed that the control region was 976 bp (GenBank accession: MF781125), COⅠ was 642 bp (MG010134) and Cyt b was 307 bp (MG010133). UPGMA and ML gene trees of the control region based on HKG + G model of 17 Baleen whales suggested that the body was among individuals in branch of Balaenoptera physalus (Fig. 4 and Fig. 5). Only a C/T transition in the control region was found between decay body and two B. physalus samples (KC572816 and KC572776) from coastal zone of California, America. The COⅠ and Cyt b sequences contained several termination codons, and the blasting results in GenBank verified that sequences of high homology were not available. The results unequivocally identified the body as B. physalus and its geographical origin was the North Pacific. Moreover, COⅠ and Cyt b in the body could be nuclear mitochondrial pseudogenes.

Abstract:During a field expedition on September 28 of 2016, we photographed one troop of about 20﹣30 Macaques in Simuzha Scenic Spot, Cona, Tibet Autonomous Region, China (27°49′34.89″N, 91°43′44.77″E, alt. 2 830 m). Their photographs and relative tail lengths are compared with that of the potiential sympatric Macaca species and from the literatures, and be sure that these macaques are Macaca munzala, which was newly discovered Macaca species in Tawang in 2005. This is the first record of this species in China except the disputed area occupied by India.

Abstract:The nemertean genus Ototyphlonemertes includes small and slender nemerteans that inhabit the interstitial spaces of marine sands. They are known to occur all over the world, but have not been reported from China seas. Recent years, we collected Ototyphlonemertes specimens from intertidal coarse sand in Qingdao (36o03′N, 120o21′E). After being relaxed with 7.5% MgCl2, specimens were observed and photographed under microscope. Two species, O. dolichobasis Kajihara, 2007 (Fig. 1) and O. martynovi Chernyshev, 1993 (Fig. 2), were identified by morphological studies. Present specimens of O. dolichobasis, which was characterized by an extremely long stylet basis and was previously known only from its type locality (Otsuchi Bay, Iwate Prefecture, Japan), have a larger body size, and a smaller central stylet and basis than those in the original description, but the ratios of “basis length / basis width” and “central stylet length / basis length” are the same as in the original description. Except being smaller in body size, the present O. martynovi specimens are well accordant to the original description. The head bristle formula of present specimens, like that of the type specimens from Peter the Great Bay, is different from O. cf. martinovi from Van Phong Bay, Vietnam. Present specimens are deposited at Institute of Evolution and Marine Biodiversity, Ocean University of China.

Abstract:In this study, we for the first time carried out Sika Deer (Cervus nippon) somatic cell culture in the space by using self-developed olyvinylidene fluoride (PVDF) membrane culture method, and analyzed the biological characteristic changes of these cells, including cell growth curve and karyotype characteristics. Ear skin tissue was collected for primary adherent culture of somatic cells, and these cells were then subcultured after trypsin digestion. The cells were cryopreserved by using a traditional procedure cooling system. The cell number was counted every day in the 24-well plates and the cell growth curve was made by applying Excel 97-2003. Non-linear regression analysis subroutine Logistic curve model from the IBM SPSS Statistics 23 statistical software was used to analyze growth curve fitting degree, and normal chromosome specimen preparation technology and cytogenetic workstation (AI) software were used to analyze chromosome karyotype of space-treated cells. The results showed that new technology of developed PVDF membrane for cell culture (Fig. 1) could effectively maintain the survival of the C. nippon somatic cells in the space and the cells successfully proliferated after recovery. The somatic cells were still showing normal fibroblast morphology after the space-mutagenesis (Fig. 2). The growth curve of somatic cells showed “S” model which was similar to that of control somatic cells. The space-treated group of cultured somatic cells entered the logarithmic growth phase by 4 days after the cultivation, while it took 2 days in the control group (Fig. 3). The rates of fitting growth curve were similar between two the groups, however, the cell inflexion time and cell volume of somatic cells were significantly changed in space-treated cells (Table 1). Karyotype analysis showed that the chromosome number was still 2n = 66 (XX), and chromosome morphology was normal, 32 of which were autosomes and one pair of sex chromosome (Fig. 4). This study established a new technology of somatic cell culture for domestic animals in the space condition, and also analyzed the effect of the space conditions on biological characteristics of C. nippon somatic cells, which provided some basic and important information and technique for future studies of animal genetics.

Abstract:Przewalski′s Gazelle (Procapra przewalskii) and Tibetan Antelope (Pantholops hodgsonii) are the first-class protected animals in China, but the digestive system structure of these two species has not been reported. We anatomized 6 Przewalski′s Gazelles and 2 Tibetan Antelopes which were dead in the wild and described their digestive systems in details. We measured the length of alimentary tract and volume of the stomach (Table 1) and analyzed percentages of various segments of the intestine of gazelles (or antelopes). The schematic diagrams of the digestive system (Fig. 1 and 2) were drawn by CorelDraw X6. Rumen of Przewalski′s gazelle occupied a large proportion of total stomach volume, and the average value was 96.05%, with 79.25% and 66.67% respectively in adult and young Tibetan Antelopes. The length of small intestine accounted for 68.42% of total length of the intestine in young Przewalski′s Gazelle and 66.48% in adult Przewalski′s Gazelle; the percentages in young and adult Tibetan Antelopes were 74.67% and 83.52% respectively. Descriptions of antelopes′ digestive system have provided basic data for further understanding and deep study of the environmental adaption of digestive system in Przewalski′s Gazelle and Tibetan Antelope.

Abstract:This work was conducted to study the histological properties and distribution of mucous cells in digestive tract by conventional paraffin section, H.E and AB-PAS (Alician blue and periodic acid Schiff reagent, Alician blue at pH 2.5) staining in Carcinoscorpius rotundicauda. The digestive tract shaped as a tube with branch structure in the zone of midgut (Fig. 1). H.E staining showed that esophagus, stomach, pyloric stomach, midgut and hindgut of this species showed general structure of digestive tract. From inner to outer of the digestive tube were mucosa, submucosa, muscularis, and serosa layer. A short and tubular esophagus was featured by thin muscularis but thick deeply chitin layer (Fig. 2a, b); Stomach was expanded with thick longitudinal folds and chitin layer coating in the mucosa surface (Fig. 2c, d, e). There pyloric stomachs trapped into midgut with both thin but recognizable muscularis and chitin layer (Fig. 2f, g); there was no remarkable difference in structure between midgut and hindgut, they both had numbers of mucosa fold and mucous cells locating in epithelium, but there was no chitin layer (Fig. 2h﹣l). Results of AB-PAS staining showed that there existed typeⅠ and typeⅡ mucous cells in digestive tract and their distribution characteristics were different in number and shape through the digestive tract. No mucous cell was detected in esophagus and pyloric stomach, but typeⅡ mucous cells were found in submucosa layer of stomach (Fig. 3a). Mucous cells distributed in mudgut and hindgut were superior in quantity than in stomach to some extent, especially in gut zone near pyloric stomach. The majority of mucous cells was TypeⅡ in midgut, mainly distributed in submucosa layer and mucosa epithelium (Fig. 3b, c). TypeⅠ mucous cells were found in submucosa layer and numerous typeⅡ mucous cells were found in mucosa epithelium of hindgut (Fig. 3d﹣f). The histological characteristics of digestive tract of C. roundicauda reveal that the functions may differ in different parts of digestive tract, and it may indicate a coordination between feeding habit and digestive tract structural evolution in C. rotundicauda.

Abstract:This study was conducted to investigate the effects of dietary supplementation of Haematococcus pluvialis powder on survival, weight gain and biochemical composition of adult male Chinese Mitten Crab (Eriocheir sinensis). Four isonitrogenous and isolipidic fattening diets were formulated by adding powder of H. pluvialis in 0, 0.2%, 0.4% and 0.6% in weight to feed the adult male E. sinensis for 60 days (defined as D1﹣D4) (Table 1). The survival rate, weight gain rate, specific growth rate and condition factor were measured and the proximate composition, fatty acid profile and carotenoid content in the tissues of male E. sinensis were detected. The experimental data was statistically analyzed with variance analysis. The results showed as the followings: (1) dietary supplementation powder of H. pluvialis had no significant effects on the survival rate, weight gain rate, specific growth rate and condition factor of adult male E. sinensis (P > 0.05) (Table 2). (2) The highest contents of protein was detected in the gonad and hepatopancreas lipid in the treatment D3, while the content of moisture in the muscle showed an increasing trend with the increasing of the powder of H. pluvialis in the diet (P < 0.05) (Table 3). (3) An increasing trend was found for the fatty acid C22:0 in the gonad with increasing powder of H. pluvialis powder in the diet, while the highest contents of C16:1n7, C18:2n6 and C18:3n3 were detected in the treatment of D4, D3 and D1, respectively (P < 0.05) (Table 4). (4) Individuals in treatment D2 had the highest contents of C14:0, C18:0 and C18:1n7 in the hepatopancreas, while the highest contents of C18:1n9 and total monounsaturated fatty acids (∑ MUFA) were detected in treatment D4 (P < 0.05) (Table 5). (5) The highest C14:0 was found in the muscle for individuals in treatment D1, while individuals in treatment D2 showed the highest C14:1n5 and C20:2n6 (P < 0.05), the crabs in treat D3 had the highest percentage of C16:0 and DHA/EPA ratio in their muscle, while the highest contents of C22:6n3, total polyunsaturated fatty acids (∑ PUFA), as well as ∑ n-3 PUFA and total highly unsaturated fatty acids (∑ HUFA) were detected in D4 treatment (P < 0.05) (Table 6). (6) The contents of astaxanthin and β-carotene in the hepatopancreas as well as astaxanthin, lutein and zeaxanthin in the carapace increased significantly with increase of H. pluvialis powder in the diet (P < 0.05) (Table 7). In conclusion, dietary supplementation of powder H. p luvialis had no significant effects on the survival rate, weight gain rate and condition factor for adult male E. sinensis, while increased the contents of gonadal protein and muscle total polyunsaturated fatty acids as well as the carotenoid contents in the hepatopancreas and carapace. These results suggested that the optimal diet was with powder of H. pluvialis at approximately 0.4 %.

Abstract:China is rich in the resources of laughingthrushes, with 40 species, 28 in Garrulax and 12 in Trochalopteron. In this paper, we summarized their distribution (Fig. 1), as well as studies on ecology, such as breeding ecology, sexual dimorphism, mating pattern, thermoregulation, adaptive thermogenesis and sing behaviours, in order to provide reference for the further research in the future.

Abstract:Mammalian hibernation is a state of seasonal heterothermy and survival strategy. The hibernation consists of long periods of deep torpor interspersed by brief interbout arousal periods. During torpor, metabolic rate is profoundly depressed and core body temperature decreases to near ambient temperature. However, these functions are promptly restored to the basic level upon arousal, and a quick restoration of blood flow accompanied by an increased mitochondrial respiration and oxygen usage results in elevated generation of reactive oxygen species (ROS) in mammals. However, no oxidative damage is found in the tissues of hibernators during a prolonged period of hibernation. Small mammalian hibernators have, therefore, been considered to be the best model of the anti-oxidative damage. In this paper, the adaptation mechanisms of oxidative stress to hibernating mammals are reviewed with respect to the generation of oxidative stress, the source of ROS and antioxidant defense.

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