Abstract:From October 2012 to May 2015, we surveyed the population of Fran?ois′ Langurs (Trachypithecus francoisi) in and nearby Mayanghe National Nature Reserve in Guizhou Province, China. Our methodologies included direct counting of langur groups and individuals along rivers and roads and at their sleeping sites, survey of local people through questionnaires and interviews, as well as the incorporation of published and unpublished data. We located a total of 72 langur groups (including 2 groups outside of the nature reserve) including 47 groups confirmed by direct counts, 8 groups by sleeping sites observations, 9 groups by questionnaires and interviews of local people with sleeping sites observations, and 8 groups incorporated from published and unpublished data. Based on a mean group size of 7.7 ± 2.9 individuals (n = 25), we estimated the langur population at Mayanghe Reserve to be approximately 554 individuals. Specifically, 41 groups (315 individuals) were found in Liangqiao area, 16 groups (123 individuals) in Gongxikou area and 15 groups (116 individuals) in Wuchuan area. The Fran?ois langurs were recorded at the mid to downstream of the Mayanghe River area, Juchishan area, Lanzihe area, Hongduhe in Gongxikou area, north Wuchuan area, south Wuchuan area and a few places such as Chanxihe, Anxi, Liangqiao, and Yintongzi. Comparing this current study to the previous one (1988﹣1989), the number of groups varied from 12 to 14﹣18 at the mid to downstream of Mayanghe River area, 10 to 9﹣13 at Lanzihe area, and 16 to 8﹣11 at Juchishan area. The main distribution area of Fran?ois′ langurs was along three major rivers: Mayanghe River, Hongdu River and Lanzi River; especially along river banks and on cliffs where vegetation occurred. Furthermore, the majority of our sightings (67 group records or 93.1%) fell within a 1 500 m radius from the center of the nearest rivers or tributaries. Thus, to ensure the species survival in Mayanghe Reserve and to reduce human-langur conflict, we recommend improved protection and restoration of natural vegetation along river banks, especially in areas densely populated by humans.

Abstract:Striped Field Mouse (Apodemus agrarius) distributes in Guizhou Province broadly, which is one of the key pest rodents in farmland. To better understand reproduction characteristics of the Striped Field Mouse population in Guizhou Province, we monitored the mouse populations in Yuqing, Xifeng, Cengong, Kaili, Leishan, Duyun, Wengan, Dafang counties from 1984 to 2014 (Table 1), assessed the reproduction parameters including sex ratio, pregnant rate of females, average fetuses and percent of males with prominent testicle, and analyzed the pattern of monthly variation of Striped Field Mouse in Guizhou. We totally trapped 20 113 mouse and made observation. There were no differences in the reproduction index between geographical populations except for average fetuses (Table 2). A two-peak type of breeding season was revealed by the monthly variation of female pregnant rate and percent of males with prominent testicle (Fig. 4). The highest proportion of pregant females occurred in spring (from April to May) and in the end of summer to beginning of autumn (from August to September) (Fig. 3). The litter sizes varied from 2 to 10, with common size of 4 to 7 (Fig. 5). The average litter size varied monthly (Fig. 6). Generally, pregnant females were present at two peaks, one in spring and one in autumn. The proportion of pregant femaled was lowest in winter (from December to February of next year). The population abundance reached the peak in June (Fig. 1).

Abstract:The Bearded Vulture (Gypaetus barbatus) is a cliff-nesting Accipitrinae bird, its territoriality is associated with the resources necessary for breeding and survival. This paper presents data of 7 breeding sites of the Bearded Vulture in the Qinghai-Tibetan Plateau collected from 1991 to 1993 and from 2010 to 2013. In total, 393 h of observation over 90 days were carried out during the investigation. Cliff-nesting birds were recorded by direct observation in 12 home ranges (a radius of 500 m away from the nests) of the Bearded Vulture (Table 1). According to our observation, 23 species of birds could arouse vigilance of the Bearded Vulture, with 11 species being nesting in their home ranges, such as Himalayan Griffon (Gyps himalayensis) ( > 107 nests), Saker Falcon (Felco cherrug) (8 nests), Golden Eagle (Aquila chrysaetos) (3 nests), Upland Buzzard (Buteo hemilasius) (3 nests) and Long-eared Owl (Asio otus) (1 nest), 3 species of Corvidae (Pyrrhocorax pyrrhocorax, P. graculus and Corvus macrorhynchos) (5 nests), Pigeons (Columba leuconota and C. rupestris) ( > 96 nests) and Bar-headed Goose (Anser indicus) (1 nests). For the total recorded 1 298 viligances caused by the 23 species, 65 interspecific interactions were observed between 12 species of birds and the Bearded Vultures, and the highest interspecific interaction frequency was the birds of Corvidae and the Bearded Vultures (34 times, 53.31% of all 12 species of birds) (Table 2). We also observed once that the Golden Eagle gave up its prey owing to the driving behavior of a Bearded Vulture in its home range. The preliminary knowledge of interspecific relationship is as following: (1) The nests of Bearded Vulture adjacent to Himalayan Griffon′s community nests, showed a relatively shorter inter-nest distance, which reduced from a range of 35.84 km (25.10﹣47.30 km, n = 6) to 7.39 km (6.22﹣8.58 km, n = 5); (2) The inter-nest distance of Saker Falcons nests in home ranges of Bearded Vulture could be only 150﹣360 m (n = 4), much shorter than ones in eastern China (1.71﹣2.00 km) and western China (the nearest distance is 2.50 km, usually from 4 to 8 km or farther); (3) As the importance of Himalayan Griffon in the local traditional culture and customs, for the nests of Bearded Vultures adjacent to the Himalayan Griffon′s community nests, the adverse effects of human disturbance on theie breeding success were reduced to a great exten; (4) When the Bearded Vultures temporarily left their nests (either for foraging or by human disturbance), the existence of Corvidae and raptorial birds (especially the Saker Falcons), could increase the possibility of their eggs and chicks being preyed; (5) During the chick-rearing period (especially the first month of chick-rearing), the lost preies of small dead animals from the raptorial birds nesting in Bearded Vulture′s home range or in the vicinity area might partly play an important supplementary role for feeding the youngs of Bearded Vulture.

Abstract:Nestling behavior and nestling growth of Yellow-bellied Prinia (Prinia flaviventris) were observed in Zhaoqing, Guangdong, China, from March to September, 2007. In this study, we recorded the food consume, nestling food composition and nestling measurements of Yellow-bellied Prinia to investigate its parental investment and nestling development pattern. The feeding behavior was observed through the window of hidden tent in a distance within 5 m from the nest. Nestlings were marked with colored, odorless pen on their backs when younger than 10 days old, and on their tarsus when nestlings were 10, 11 and 12 days old. Bivariate correlation was employed to analyze the relationship between nestling age and types of food fed by their parents, and the correlation matrix of nestling measurements. Also, logistic growth curve was used to fit the nestling growth data, and the k value of each regression was performed a significance test. Our results showed that: 1) Nestlings were fed by both parents, with the nestling period of 11.9 ± 0.4 d (n = 7). When nestlings were less than 7 days old, parents spent more time on warming nestlings in the morning and evening; 2) Feeding frequency and the types of food increased as nestlings grew older, with nestling age positively correlated with parental feeding frequency (r = 0.995, P < 0.01); 3) Nestlings were fed with arthropods only, with the spiders being the most (40.95%) and others including larvas, Orthoptera, Lepidoptera, Coleoptera, Odonata and so on (Fig. 1); 4) The k value of growth curve for body weight (Fig. 2) was the maximum value (Table 1), and differed significantly from other k values (P < 0.05) of length growth curves (Fig. 3); 5) All the correlation coefficients among body measurements (body weight, body length, tail length, wing length, culmen length, gap length, the third primary feather (abbreviated as ‘P3’ below) and length of tarsus and claw) of nestlings were larger than 0.9 (P < 0.01, Table 2), indicating that body parameters could be replaced each other at some extent; 6) The most important functions of the nestling organ developed preferentially (Table 3), in accordance with the energy allocation hypothesis. In summary, Yellow-bellied Prinia had relatively higher feeding frequencies and k values, possibly due to rich food source in our study area, and/or an adaption to highly variable nest habitats.

Abstract:成年扬子鳄（Alligator sinensis）的声信号已经有过深入的研究，然而其声音的个体发育，特别是幼鳄出壳前后鸣声的时空特征至今还不为人所知。为了解扬子鳄声音的发育过程，记录和分析了21只扬子鳄幼鳄的260个叫声，结果表明，幼鳄出壳前后的鸣声具有相似的特征，即开始向上的频率调节和较长的向下的频率调节。出壳前和出壳后的两组叫声均包括持续叠加的多组谐波，并且绝大多数的鸣声都是在第3谐波显示了最强的能量。分析了出壳前后鸣声的9个参数，即总时长、向上调频的时长、向下调频的时长、基频的起始频率、最高频率、结束频率、向上调频的倾斜度、向下调频的倾斜度、主频。除了向上调频的时长，出壳前后鸣声的其他时长参数均没有表现出显著差异。与之相反的是，出壳前后鸣声的频谱参数，特别是基频的最高频率和主频均表现出显著差异（P < 0.05）。出壳前鸣声基频的最高频率为（573 ± 103）Hz，出壳后的声音最高频率（460 ± 52）Hz，出壳前比出壳后高113 Hz；出壳前的声音主频（1 359 ± 229）Hz，出壳后的声音主频（1 123 ± 216）Hz，前者比后者高236 Hz。幼鳄发出主频较高的鸣叫有利于吸引附近母鳄的注意同时避免天敌的捕食。

Abstract:Dayang Newt (Cynops orphicus) is an endangered species and endemic to China, there′s no any report on its reproductive ecology. The breeding season of Dayang Newt starts in the middle of May, we paired female newt with multiple males in aquariums to observe interaction of them and record duration or frequency of different sexual movements. The sexual behavior of this species resembled that of Chinese Fire-bellied Newt (C. orientalis), but male C. oriphicus had longer tail-fanning and creeping duration (25﹣37 s) than C. orientalis (10﹣20 s, Table 3), female might picks up more than one spermatophore if male deposited several in his courtship. The spermatophore shape is similar to C. orientalis. Males bit the belly after her pick-up spermatophore and she remained stationary (Fig. 1d). Female did not show any interest and even frightened in male once she was inseminated. The successful proportion of spermatophore picking-up was relatively rare (2.7%, Table 2), but slightly higher than that recorded in C. orientalis (1.1%). The competitions and interferences behaviors were found from potential male rival during courtship.

Abstract:Elongate Loach (Leptobotia elongate) is endemic to the upper Yangtze River. It has been threatened by overfishing and dam construction, and evaluated as being endangered in China Species Red List. To understand its population dynamics after the Three Gorges Reservoir was impounded, the catch and time series length-frequency data were used to estimate growth and mortality parameters, population abundance and resources utilization of Elongate Loach based on surveys in the Jiangjin section of the upper Yangtze River conducted in 2007﹣2009 (Fig. 1). Elongate Loach ranged from 76 to 480 mm in length and 5 to 2002 g in weight, with an average length of 158.7 ± 54.8 mm and an average weight of 72.4 ± 148.8 g (n = 277). The length group of 90﹣210 mm dominated the catches (77.9% of the total number) (Fig. 2). The length (L, mm)-weight (W, g) relationship of Elongate Loach was well-fit with a power function, W = 7.28 × 10﹣6 L3.09 (R2 = 0.95, P < 0.01, n = 277) (Fig. 3). Asymptotic length (L∞) and growth constant (k) were estimated using length frequency data as 555 mm and 0.17/a, respectively. Natural mortality was estimated as 0.37 using the empirical formula proposed by Pauly. The total mortality was estimated using a length-converted catch curve analysis as 1.23 (Table 1). The exploitation rate observed in the Jiangjin section was 0.70, higher than the estimated maximum exploitation rate (0.43), which indicated that it was overfished for Elongate Loach (Fig. 4). Population abundance of Elongate Loach in the Jiangjin section estimated by length-structured virtual population analysis was 2 544 ind/km (0.75 t/km) in 2007, 2 405 ind/km (0.42 t/km) in 2008, and 7 245 ind/km (1.63 t/km) in 2009, respectively, with an average of 4 065 ind/km (0.93 t/km) (Fig. 5). Compared with the earlier studies (Table 2), a new maximum length (480 mm) of Elongate Loach was recorded; the estimated total mortality and exploitation rate of Elongate Loach in this study were higher, which was likely caused by the higher fishing effort in the Jiangjin section. Long-term population dynamics monitoring, closed fishing, and artificial enhancement and releasing were suggested to improve the resources.

Abstract:In order to explore the function of β-carotene-15, 15′-monooxygenase 1 (BCMO1) in vitamin A (VA) metabolic balance in Yak (Bos mutus), the expression levels of BCMO1 protein were studied in Yak tissues at different nutrition periods. Three healthy adult males of 3 years old were selected for samples collection at July 2013 (nutrition-rich period) and March 2014 (nutrition-deficiency period) at Tongpu town of Wulan County in Qinghai Province. Western blotting were used to detect the expression of BCMO1 protein in eight tissues including liver, lung, kidney, muscle, duodenum, ileum, jejunum and rumen. The different expression levels of BCMO1 protein in different tissues at the same nutrition stage was compared using multiple comparisons and the difference of the expression level at different nutrition periods for the same tissue was compared using T test. As shown in Fig. 1 and Fig. 2, there was the highest expression level of BCMO1 protein in duodenum among all tissues (P < 0.05), and the expression level in ileum and jejunum tissue was higher than that of kidney and rumen (P < 0.05). BCMO1 protein expression level was the lowest in liver among all tissues during the nutrition-deficiency period. During the nutrient-rich period, liver tissue showed the highest level of BCMO1 protein among all tissues (P < 0.05). As shown in Fig. 2 and Table 1, the expression of BCMO1 protein in the liver was 0.919 ± 0.228 at nutrition-rich period, which was significantly higher than that at nutrient-deficiency period (P < 0.05). However, the expression levels of BCMO1 protein in duodenum, jejunum, kidney, muscle and rumen were 0.811 ± 0.134, 0.336 ± 0.255, 0.739 ± 0.404, 0.619 ± 0.092 respectively at nutrition-rich period, which were lower than those at nutrition-deficiency period (P < 0.05). The BCMO1 expression levels in lung were 1.128 ± 0.407 and 1.083 ± 0.232, and its expression levels in ileum were 0.466 ± 0.337 and 0.517 ± 0.280 at nutrition-rich and nutrition-deficiency periods, respectively. There was no significant difference between two different nutritional periods for BCMO1 expression in two tissues (P > 0.05). The expression of BCMO1 protein was lower at nutrition-rich period than that at the nutrition-deficiency period for most tissues, which indicated that sufficient content of vitamin A in tissues might inhibit expression of BCMO1, as the contents of β-carotene and vitamin A were adequate in Yak tissues at nutrition-rich time. It is very important to maintain and regulate vitamin A metabolic balance in Yak at different nutrition periods.

Abstract:This study aims to explore the influence of fragile X retardation 1 (FMR1) gene knockout on biological characteristics of C57BL/6 (Mus musculus domesticus) mice. Blood physiological and biochemical indexes of FMR1 knockout mice and C57BL/6 mice of 8﹣10 weeks old were respectively examined using animal automatic blood cell analyzer and automatic biochemical detection. The comparison of difference between groups was analyzed with independent-samples T test method using SPSS 19.0. The blood routine indexes of C57BL/6 FMR1 KO male and female mice, such as neutrophilic granulocyte (MEUT#), neutrophilic granulocyte percentage (MEUT) and lymphocyte percentage (LY), had significant difference compared to C57BL/6 mice (P < 0.05); for males, the number of red blood cells (RBC), hamoglobin (HGB), hematocrit value (HCT) and mean corpuscular hemoglobin concentration (MCHC) showed significant difference compared to C57BL/6 mice (P < 0.05); for females, the number of white blood cells (WBC) and lymphocytes (LY#) showed significant difference compared to C57BL/6 mice (P < 0.05); the non-sex factors including red blood cells (RBC), hematocrit value (HCT), neutrophilic granulocytes (MEUT#), neutrophilic granulocyte percentage (MEUT) and lymphocyte percentage (LY) showed significant difference compared to C57BL/6 mice (P < 0.05). But there was no significant difference in the number of platelets (PLT), mean platelet volume (MPV), plateletcrit (PCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), red blood cell distribution width-standard deviation (RDW-SD), red blood cell distribution width-coefficient of variance (RDW-CV), platelet distribution width (PDW), platelet-large cell rate (P-LCR), monocytes (MONO #), monocytes percentage (MONO), eosnophils (EO#), eosnophils percentage (EO), basophilic granulocytes (BASO#) and basophilic granulocyte percentage (BASO) between C57BL/6 FMR1 KO mice and C57BL/6 mice (P > 0.05) (Table 1). Serum biochemical indexes of C57BL/6 FMR1 KO mice, such as the AST/ALT (AS/AL) in males had significant difference compared to C57BL/6 mice (P < 0.05); the creatinine (CR), creatine phosphokinase (CK) and electrolyte Ca2+ in females had significant difference compared to C57BL/6 mice (P < 0.05); the non-sex factors including AST/ALT (AS/AL), creatinine (CR) and creatine phosphokinase (CK) had significant difference compared to C57BL/6 mice (P < 0.05). No significant difference was found in albumin (ALB), globulin (GLB), ALB/GLB (A/G), alkaline phosphatase (ALP), glutamyltransferase (GGT), total protein (TPROT), alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), uric acid (UA), cholesterol (CHO), triglyceride (TG), glucose (GLU), lactic dehydrogenase (LDH), osmotic pressure, K+, Na+, Cl￣, Mg2+ and P3+ (P > 0.05) (Table 2). Thus, the results suggest that FMR1 gene knockout can influence some blood physiological and serum biochemical values in mice. These results will provide the experimental basis for researching and using C57BL/6 FMR1 KO mouse model in the future.

Abstract:To investigate the effects of artificially induced hibernation on the consumption of macronutrients in the blood and tissues in Asiatic Toads, Bufo gargarizans, hibernation was artificially induced for 56 days, and the body mass, organic indexes of heart, liver and gastrocnemius muscle, as well as the concentrations of glucose, total protein and total cholesterol in blood, the contents of glycogen in the hepatic tissue and skeletal muscle, and protein in the skeletal muscle and myocardium were measured on the day before hibernation and the day of 1, 3, 7, 14, 28, 42 and 56 post hibernation. Results showed that, compared with the pre-hibernation, the body mass did not change significantly during hibernation, and there was not significant sexual difference (Plot a1 and a2 in Fig. 1). The organic indexes of the heart, liver and gastrocnemius muscle in male toads were significantly higher than those in females (P < 0.01) (Plot b2, c2 and d2 in Fig. 1), while both the males and females showed no significant change in organic indexes during hibernation (Plot b3, c3 and d3 in Fig. 1). The blood glucose concentration decreased significantly after day 42 (P < 0.01) (Plot a1 in Fig. 2), the concentration of total protein decreased significantly (P < 0.05) on day 56 (Plot b1 in Fig. 2), but total cholesterol did not change significantly (Plot c1 in Fig. 2). The blood macronutrients did not show significant sexual difference (Plot a2, b2 and c2 in Fig. 2). The hepatic glycogen decreased significantly after day 42 of hibernation (P < 0.01) (Plot a1 in Fig. 3), and the muscular glycogen decreased significantly after day 1 (P < 0.05) (Plot b1 in Fig. 3), but the protein contents in the skeletal muscle and myocardium did not show significant differences (Plot c1 and d1 in Fig. 3). The tissue macronutrients showed no significant sexual difference (Plot a2, b2, c2 and d2 in Fig. 3). The contents of carbohydrate in the blood and tissue decreased first, and the blood protein decreased significantly only after deep hibernation, but the macronutrients in the blood and tissue could keep a relatively stable state within 1 month. It might be one of the major physiological mechanisms to adapt to hibernation in toads.

Abstract:The effect of alkalinity (0, 2 and 4 g/L NaHCO3) stress on morphology of gill ionocytes in Nile Tilapia (Oreochromis niloticus) was studied by scanning electron microscopy, and four HCO3- transporters including carbonic anhydrase (CAⅡ, CAⅣ), Na+/HCO3- cotransporter (SLC4A4), and Cl-/HCO3- exchanger (SLC26A6) in gill, kidney and intestine were also observed by immunohistochemistry. The surface scanning of the gills showed that ionocytes cells were distributed close to the inter-lamellar regions of gill filaments (Fig. 1). According to the apical shape and size, the ionocytes cells could be divided into four subtypes, subtype Ⅰ, subtype Ⅱ, subtype Ⅲ and subtype Ⅳ (Fig. 2). The apical size of each subtype of ionocytes was positively correlated with the alkalinity stress strength, and the size of subtype Ⅲ cells was changed most obviously (P < 0.01) (Table 1); The ionocytes number increased significantly with the increase of alkalinity stress strength, and the number of subtype Ⅲ cells was increased most significantly (P < 0.01) (Table 1). The results of immunohistochemistry showed that CAⅡ, CAⅣ, SLC4A4 and SLC26A6 were expressed in gill and kidney of O. niloticus at fresh water and alkaline water (Fig. 3, Fig. 4). With the increase of alkalinity, the positive reactions became stronger (Table 2). However, no positive reaction was detected in intestinal at fresh water and alkaline water (Fig. 5). This result suggest that O. niloticus may change the quantity and morphological structure of ionocytes to adapt to alkaline environment, and that gill and kidney may play a key role in alkalinity regulation.

Abstract:Orange-spotted Grouper (Epinephelus coioides) which inhabits subtropical and tropical areas, has become a very popular species of marine teleost currently being cultured in China. Metabolic rate is the most fundamental biological rate as it represents the rate of energy uptake, transformation and allocation. Oxygen consumption is a widely studied indicator of metabolic rate, and measurement of oxygen consumption rate is often used to examine energy utilization and stress in aquatic animals. The aim of the present study is to investigate the effects of temperature (21℃, 24℃, 27℃, 30℃ and 33℃), pH (6.5, 7.0, 7.5, 8.0 and 8.5) and body wet weight (mean values:15.64 g, 35.80 g, 65.67 g and 95.93 g) on oxygen consumption rate (MO2), ammonia excretion rate (MTAN) of Orange-spotted Grouper juveniles cultured in low-salt water (salinity 12) by the intermittent flow respirometry system with ecological methods in laboratory. Data were analyzed by using the one-way ANOVA, followed by Ducan test. All analyses were performed with a significance level of P < 0.05. The results showed that temperature strongly affected both MO2 and MTAN. The MO2 and MTAN increased significantly (P < 0.05) with an increase in temperature from 21℃ to 33℃ (Table 1). The linear equation between the MO2, MTAN and temperature (T) were established (MO2 = 6.0826T﹣8.9704, R2 = 0.9127; MTAN = 0.2248T﹣0.7731, R2 = 0.7792) (Fig. 2). Over the entire experimental temperature range (21﹣33℃), the respiration temperature coefficient Q10 and excretion temperature coefficient Q10 were 1.51, 1.54, respectively, and the lowest values were found between 27 and 30℃. The optimal temperature for the juvenile lied between 27 and 30℃ (Table 2). Both MO2 and MTAN were significantly affected by pH (P < 0.05) (Table 1), being increased first then decreased with pH increase. The relationship between MO2, MTAN and pH was modeled by a quadratic equation (MO2 = ﹣15.241ApH2 + 234.98ApH ﹣737.42, R2 = 0.7888; MTAN =﹣1.1477ApH2 + 18.073ApH﹣65.369, R2 = 0.7557) (Fig. 2). Both MO2 and MTAN decreased significantly with increased body wet weight (P < 0.05) (Table 1), and their relationship with body wet weight (W) could be represented by power equations (MO2 = 310.61W ﹣0.1972, R2 = 0.8653; MTAN = 9.9167W ﹣0.2043, R2 = 0.8257). The oxygen consumption (RO2) and ammonia excretion (RTAN) increased with increase in body mass, and their relationship with body wet weight was modeled by power equations (RO2 = 0.3106W 0.8028, R2 = 0.9907; RTAN = 0.0099W 0.7957, R2 = 0.9863). The averages of oxygen-nitrogen ratio (O︰N) at different temperatures, pH values and body wet weights were 25.90, 28.65 and 28.19, respectively (Table 1), which indicated that Orange-spotted Grouper juvenile cultured in low- salt water utilized protein-lipid dominated metabolism.

Abstract:Hypoxia occurs in the natural and aquaculture water environment frequently. Golden Pompano (Trachinotus ovatus) is a hypoxia-sensitive fish, so we employed (31.59 ± 3.01g) juvenile golden pompano to study the effect of acute and chronic hypoxia stress on the liver by physiological and histological methods. Individuals were exposed to 3﹣24 h of acute or 14 d of chronic hypoxia stress at room temperature. The histological changes in the liver were observed to analyze the tissue injury. Catalase (CAT), superoxide dismutase (SOD), glutathione (GSH) and malonic dialdehyde (MDA) activities were measured to determine the antioxidant effects in the liver tissue. Data were statistically analyzed. The liver tissue gradually showed vacuoles, expanded blood sinus, and blurred hepatic lobule structure during acute hypoxia. At 24 h acute hypoxia, liver tissue even showed cell fusion and local cell necrosis (Fig. 1e). At 14 d chronic hypoxia, liver tissue showed local cell necrosis and vacuolation inside the cells (Fig. 1f). Cell structure was decentralized, intercellular connection was divided, cell membrane was dissolved, organelles were disrupted, nucleus was decomposed, blood sinus was enlarged, while only rough endoplasmic reticulum was evidently observed (Fig. 2c, d). In acute hypoxia, CAT activity was continuously increased, SOD and GSH activities was recovered after increase (P < 0.05). In chronic hypoxia stress, SOD, GSH activities were significantly increased (P < 0.05), while CAT activity was significantly decreased (P < 0.05) (Fig. 3a﹣c). MDA activity firstly was increased and then recovered in acute hypoxia, but increased significantly (P < 0.05) at 14 d of hypoxia (Fig. 3d). The results show that golden pompano liver tissue structure is seriously damaged and is in severe oxidative stress under hypoxia stress. Hypoxia stress in chronic hypoxia is more serious than that in acute hypoxia.

Abstract:Molting is an essential biological process for the growth and development of crustaceans, however, the molecular mechanism of molting on growth regulation is little known. In this study, molting and growth performance were individually observed under the indoor laboratory condition, and gene expression level of two molting-related genes (ecdysteroid receptor gene, EcR; retinoid X receptor, RXR), and one growth related gene (myostatin, MSTN) were investigated using quantitative RT-PCR method (Table 1 and Fig. 4). A total of 60 juvenile crabs in each male and female groups were reared in 120 water closets of 60 L with 1 individual in each closet for two molting times. The growth traits (body weight, carapace length, carapace width, and merus length of second foot) were measured for each crab at the initial stocking stage, on the fifth day after the first molting and the second day after the second molting. Meantime, muscle and hepatoparncreas tissues were immediately collected and stored in ﹣80℃ freezer after growth traits measurements at the second molting for further gene expression analysis. The correlation analysis was conducted between the expression level of the three genes (EcR, RXR and MSTN) and growth traits using SPSS software. A sharp gaining of body weight was found after molting, followed by a slow and continuous increment in post-molting stage (Fig. 1, 2). Condition factor that could be used as an index indicating the next molting was initiated when it accumulated over 60% (Fig. 3). There was no significant correlation between the conditional factor and morphological character (Table 2). Significant negative correlation was found between MSTN expression level and the growth rate of carapace length (r =﹣0.450, P < 0.05) and carapace width (r =﹣0.410, P < 0.05), but significantly positive correlation was detected between MSTN expression level and condition factor (r = 0.450, P < 0.05) (Table 3). There was no significant correlation between the EcR and RXR expression levels and the rate of weight increment, carapace length, and carapace width increment. Generally, higher expression of MSTN indicated lower rate of weight increment, and higher expression of EcR and RXR indicated higher rate of weight increment (Fig. 5, 6). This research indicates that a regular pattern on the growth performance, and condition factor measuring nutrient substance content could be used as an indicator showing the initiation of the next molting, and the gene expression of EcR, RXR, and MSTN are correlated with the growth traits in the juvenile Chinese Mitten Crab.

Abstract:Five isonitrogenous and isolipidic fattening diets were formulated by blending vegetable oils (soybean oil: rapeseed oil = 1︰1) to replace fish oil at level of 0, 25%, 50%, 75% and 100% (named as diet F1, F2, F3, F4 and F5, Table 1) to feed 5 treatments of adult male Chinese Mitten Crab (Eriocheir sinensis) for 60 days. Then, we assayed the proximate composition and fatty acid composition of the hepatopancreas, muscle and testis of each crab in the five treatments and analyzed these data by variance analysis method. We got the following results: (1) The gonads of crabs fed F1 diet had the significantly higher ash content than those fed by diet F4 and F5 (P < 0.05), but no significant difference with crabs fed diet F2 and F3; no significant differences in the contents of moisture, crude protein and crude lipid among five diet treatments (P > 0.05). The contents of moisture and ash in the hepatopancreas of crabs fed diet F1 were highest among the five treatment group, but the crude lipid was lower than other four treatments; however, no significant difference was found in the crude protein content between individuals in the five treatments (P > 0.05). Except for the crabs treated by diet F1, the crude lipid and ash contents in the muscle increased with increasing replacement level; while no significant differences were found for the rest contents in the muscle among crabs treated by five diet (P > 0.05) (Table 3). (2) The total n-6 polyunsaturated fatty acid (∑ n-6PUFA) contents in gonads of crabs increased with the increasing replacement level, while the content of total n-3 polyunsaturated fatty acid (∑ n-3PUFA) had opposite changing tendency. No significant difference was found in the total saturated fatty acid (∑ SFA), total polyunsaturated fatty acid (∑ PUFA) and total highly unsaturated fatty acid (∑ HUFA) contents in the crab gonads between all the five treatments (P > 0.05) (Table 4). (3) The highest content of ∑ n-3PUFA and ∑ HUFA was detected in the hepatopancreas of crabs treated by diet F3 although there was a significant difference between each treatment. While no significant differences were found for the ∑ PUFA and ∑ n-6PUFA contents among five treatments (P > 0.05) (Table 5). (4) There was no significant difference among the fatty acids contents in muscle except for the content of ∑ n-6PUFA which increased with elevating replacement levels (Table 6). In conclusion, except for moisture and crude protein contents, fish oil replacement by blending vegetable oils in fattening diets have significant influence on fatty acid composition in the edible tissues of adult male E. sinensis. These results suggested that the replacement level of 50% is good for the fat accumulation in the hepatopancreas and muscle of this crab.

Abstract:SNaPshot assay is a single base extension assay which, by extending a base after four primers, displays color of the base through capillary gel electrophoresis to judge the genotype of samples. We aimed to establish a SNaPshot assay to type mouse hepatitis viruses (MHV), including MHV1, MHV3, JHM, and A59. The SNaPshot assay was developed after polymerase chain reaction (PCR) using cDNA from MHV strains two pairs of universal PCR primers and four single-base primers that were designed according to the conserved sequence of the four strains. The PCR products were analyzed by capillary gel electrophoresis. The sensitivity and specificity of the SNaPshot assay were analyzed using gradient dilution method and compared with human Hepatitis B virus (HBV). The accuracy of the SNaPshot assay was analyzed using 41 murine (Mus musculus) serum samples by comparing with ELISA and sequencing the samples containing MHVs. The optimal conditions of SNaPshot assay were that T1﹣T4 primers were 0, 3, 10, and 15 Ts in modification, 4︰6︰5︰10 in concentration, and 16 bp, 19 bp, 26 bp and 31 bp in length. The sensitivity of the SNaPShot assay was 1.25 mg/L (MHV cDNA) (Fig. 3), the specificity was 100% (Fig. 4), and the accuracy was 100% (41/41, Table 3), which was confirmed by ELISA, and the sequencing data. The SNaPshot assay is a sensitive, specific, and accurate method for typing MHV1, MHV3, JHM, and A59.

Abstract:Equilibrium, freezing and thawing are three essential steps which play a critical role in semen freezing process. In order to optimize Horse (Equus caballus) sperm freezing methods, we investigated the impact of the equilibrium, freezing and thawing manipulations on sperm motility, membrane integrity and mitochondrial membrane potential after freeze-thawing. Semen was collected from 4 adult stallions, diluted in INRA82 + 5% clarified egg yolk + 3.5% combined cryoprotectant and then frozen. In excrement 1, we compared the effect of equilibration for 0, 45, 90, 120, 180, 240 min and 8 h on equine sperm cryopreservation; in experiment 2, we aimed to find a suitable height of liquid nitrogen steam for freezing; in exprement 3, we assessed the effect of different thawing procedures on post-thaw semen quality. Statistical analysis was conducted with ANOVA SPSS 13. Differences between means of parameters were subjected to an analysis of variance using the Tukey′s test, with P < 0.05 considered significant. The results showed that the equilibrium time of 120, 180 and 240 min resulted in significantly higher sperm motility and membrane integrity than the equilibrium for 0, 45, 90 min and 8 h group after thawing (Fig. 2); liquid nitrogen steam frozen at 2 cm and 4 cm from liquid nitrogen surface obtained similar results with program freezing method (Fig. 3); high temperature rapid thawing method (75℃ 7 s and 46℃ 20 s) gained a higher post-thaw motility than the conventional thawing method (37℃ 30 s) (Fig. 4). In summary, we believe that combination of equilibrium for 120﹣240 min, liquid nitrogen steam frozen at 2 cm and 4 cm from liquid nitrogen surface and high-temperature rapid thawing method (75℃ 7 s and 46℃ 20 s) can get better effect in horse sperm freezing process.

Abstract:During the breeding season from April to August in 2015, one case of White-bellied Redstart (Hodgsonius phaenicuroides) parasitized by Large Hawk-cuckoo (Cuculus sparverioides) was observed in Kuankuoshui National Nature Reserve, Guizhou, southwestern of China, and the cuckoo parasitism rate was 1/16 (n = 16). The parasitized nest, with one white Cuckoo′s egg and two Redstart′s eggs (Fig. 1), was found to be deserted by the redstart host. The Large Hawk-cuckoo egg was 6.09 g in weight and 27.73 mm × 20.20 mm in size, and two host eggs were 2.34 g and 2.40 g in weight and 20.05 mm × 14.94 mm and 19.95 mm × 15.09 mm in size, they were much different in egg mass and egg size (Table 1). Egg measurements by spectrophotometer showed that they were much difference in spectrogram. The Large Hawk-cuckoo egg was a typical non-mimetic parasitic egg because of its egg chroma and hude were much lower than those of White-bellied Redstart, however, higher in brilliance (Fig. 2).

Abstract:The Oriental Scops Owl (Otus sunia) is wide spread in South and East Asia but little was known about its reproductive feeding behaviour, particularly in China. The main objective of this study is to assess and evaluate the diet and feeding activity of the owls during reproductive season. The work was conducted in an experimental wood of Nanjing University, Nanjing, China in 2012 and 2013 by means of infrared video-recording, with each year one nest being recorded. Food types were recorded and feeding rhythm of the owls was analyzed with ANOVA and Student-Newman-Keuls post-hoc for multiple-level comparison. According to 613 records which we could identify the type of food from the videos, we found that the food items delivered to the owlets were composed of maily insects belonging to Orthoptera (62%) and Lepidoptera (18%), and non-insect invertebrates such as Araneae (8%) and Chilopoda (7%), and small terrestrial vertebrates (3%) such as rodents and geckos. The overall feeding rhythm of delivering food items by parent owls was varied significantly, with three feeding peaks at 19:00﹣20:00, 21:00﹣22:00 after sunset and 04:00﹣05:00 before sunrise (Fig. 1). With this study, we confirmed that insects are the most important food Scops Owls during breeding period and the peak time of active feeding of Scops Owls is during dusk and dawn food of Scops.

Abstract:Six specimens of odour frogs were collected from Fanjing Mountain National Nature Reserve (FMNNR) in Guizhou Province from May to July in 2015 (Table 1). The specimens were identified as Odorrana yizhangensis (Amphibia, Anura, Ranidae); and this is the new record of this species in Guizhou Province. Twenty two morphological characters of the specimens were measured (Table 2) and their habitats (Fig. 1) were also described. The maximum likelihood phylogenetic tree constructed based on 12S rRNA and 16S rRNA gene fragments showed that the six specimens were clustered with the O. yizhangensis sampled from its type locality (Fig. 2), and the genetic distance of Kamura-2-parameter model between our specimens and O. yizhangensis sample from its type locality was much smaller than that between Odorrana species (Table 3). New found of O. yizhangensis in Guizhou Province would promote taxonomic and phylogeographic studies on this species.

Abstract:Vocal communication consists of the production, transmission and perception of sounds as well as behavioral response to sounds. For most anurans, competition between males, potential mate recognition and choice by females are almost totally dependent on the vocal communication. Therefore vocal communication will be critical for the survival and reproductive success in frogs. In this paper, we reviewed previous studies on the calling characteristics of frog and the production mechanism for their callings. We summarized the functions and evolution of vocal communication in sexual selection, possible neural mechanisms and endocrine mechanisms underlying vocal production and auditory perception. Finally, we proposed some potential study fields for the vocal communication and predicted the possible prospects in the future.

Abstract:Based on the related literatures studying, this paper summarized the research progress on morphological and biological characteristics of the bdelloid rotifers with emphasis on their parthenogenesis and anhydrobiosis features as well as their mechanisms in bdelloid rotifers. The results of current researches on the bdelloid rotifers put forward new challenges to the traditional theories about classical genetics, biochemistry and physiology. These progresses also has certain reference on the safeties of “transgenic” organisms. The research reports on the new species bdelloid rotifer species and their fauna not only had rich global invertebrate biodiversity, but also provided important reference value on the improvement of the geographical distribution pattern for this group of animals. Based on the study status and its significance in academic of bdelloid rotifer, we suggest that it should strengthen the bdelloid rotifer research in domestic.

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Abstract:The Chinese Crested Tern, Thalasseus bernsteini, a seabird of eastern Asia, has far been considered a Critically Endangered (CR) species since the very beginning of quantity assessment on the endangerment of birds. Although, in the last decade years or so, only two colonies of the bird had far been reported, that is, the colony in the Taiwan Straits and the colony around the Greater Zhoushan Archipelago, respectively, the authors pointed out, in 2010, that the Northern colony of the bird might currently still “be appearing in the rim of the Yellow Sea”. Then, in 2011, new findings of the bird got revealed in Rizhao of Shandong. Quite recenly, on 7 August 2016, Chinese birders (Zhang Lin and others) found an adult bird with a bird of the year inhabiting on the coast at Rudong of Jiangsu, E China. Next day, August 8, the Korean governmental authorities officially declared that two breeding pairs of the Chinese Crested Tern observed started nesting on an island offshore the SW Korean coast in April and the whole process being well recorded. Then, from August 19 to September 5, people of Qingdao Birdwatching Society (Yu Tao, Wang Hai-bin, and others) constantly saw the bird in Jiaozhou Gulf of E Shandong, and, to be the extreme, two adults with two birds of the year on August 19﹣20 and a small flock of six adult birds on August 30 observed. It hence might be doubtlessness of current existence of the CCT′s northern colony, while, these updated new findings have also brought with them further evidence onto the jigsaw puzzle of the breeding distribution of the bird, making it now appearing relatively more completed and integrated than ever before. At last, we still would like offerring the same question, like once we did in 2010, that is, what a portion of the CCT′s world population that we can say under monitoring, or, simply to say, in our hand or our sight… 80%, 50%, 30%, or even less?

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