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07 July 2008, Volume 24 Issue 2

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The Winter Habitat and Its Characteristics of Golden Takin in the East Part of Qinling Mountains

MA Yingtai LI Zhenbin WU Xuntao SHAO Jianbin ZHAO Wenchao

2004, 24(2):  93-97. 

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The winter habitat of Golden Takin ( Budorcas taxicolor bedfordi) in Niubeiliang National Nature Reserve which locates in the east part of Qinling Mountaints were surveyed fromJan. to Apr. , 19981 Five habitat patches were identified , the total area of the winter habitat was 2 305 ha , it occupied 14.04 % of the whole reserve. The main characteristics for Golden takins to select their winter habitat were topography (with a sunny exposure , against the wind , and subalpine terrace of relatively smooth terrain) ,food resource and quality , water (sufficiency) , cover for escaping natural enemy and comparatively warmth environment provided by coniferous forest and mixed coniferous and broadleaf deciduous forest , and escape natural enemy habitat of Fargesia spathacea. The twig , leaf and skin of Abies sp. , Pinus armandii were the major winter food taken by Takins in the study area. The fargess fir (Abies sp.) was the indicator tree species for the distribution of Takins. The forest of fargess fir covers Takins from cold wind , snowing during winter. The Takins have been restricted within the habitat patches because of the habitat fragment resulted from logging. The National 210 highway is the major obstacles for takins traveling from east to west within the reserve. Habitat restoration and building corridor over the 210 highway will be the very important management strategies for the conservation of Golden takins in the east part of Qinling Mountains.

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Observation on Some Sexual Behavior of the Yangtze Finless Porpoise

WEI Zhuo WANGDing ZHANG Xianfeng WANG Kexiong CHEN Daoquan ZHAO Qingzhong KUANG Xinan GONGWeimin WANG Xiaoqiang

2004, 24(2):  98-102. 

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The finless porpoise ( Neophocaena phocaenoides) is a small-toothed cetacean species , and distributes in a narrow band along the coast of tropical and temperate Asia. The population inhabited in the Yangtze River is a unique freshwater subspecies, and is known as the Yangtze finless porpoise ( N. p. asiaeorientalis) . Its population size has been decreasing remarkably in recently years. In order to breed the animals in captivity for the purpose of conservation , 3 Yangtze finless porpoises have been kept in our aquarium for 3 to 6 years. The rearing hall is a round building that is 30 m across , with a main kidney shaped pool that is 20 m ×7 m ×315 m , an auxiliary round pool that is 10 m across and a medical pool that is 7 m across. The main pool has 7 under water viewing windows that are 2 m×115 m in size , where that animals was kept and observation was carried out. The window in the middle of the pool was chosen to be the observation site. A pre-designed sheet was used to record behaviors and other environment information. Data recorded included record date , record time , weather , air and water temperature , air pressure , pH , clarity , penis erected behavior , water layers (up , middle and low) in which it happened , the time length it maintained , which female it chased when penis was erected , and responded behavior of the females , and so on. A 35 mm camera with 80 - 300 mm lens and
a digital video camera were used to take pictures. A digital voice recorder pencil was used to record comments by the recorder , a digital recorder was used to measure water and air temperatures and pH values , a air pressure metre was used to measure air pressure , and Secchi disc was used to measure clarity of the water. In total , 120 stationary observations were carried out from December 2001 to November 2002. The total observation time is 4 800 min. In this period , 105 times behaviors of penis erected were observed. Statistical analyses of the data indicated that the male animal stretched penis out of its body every month , the duration was from 1 to 6 s with the average of 218 s ( ±SD 0.3) . This kind of behavior had 2 peaks during April to May and September to October. The behavior could happen in different water layers , and there was no significant differences between them. The pattern that the male stretched penis out of body included sidestroke erection ; bend erection ; up erection ; down erection and perpendicular eretion. This may indicate the diversity of the animals'mating patterns. The male chased adult female more often than juvenile one. We conclude that mix group is better for normal breeding behavior development of this animal. During the observation period , the range of air pressure was 1 000 - 1 025 hpa , clarity was 3 - 315 m, pH value was 711 - 7153 , and the pool was lighted naturally. The sexual behavior was not affected by these environmental factors1During peak seasons of penis erected behavior , water and air temperatures were 17 - 23 ℃, the difference between them was 0 - 4℃. These results provide a base for reproducing the Yangtze finless porpoises in captivity.

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Study on the PCR Method Specif ic for Tiger Identif ication

HUA Yuping ZHANGQiong XU Yanchun ZHENGDong

2004, 24(2):  103-108. 

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For develpment a PCR specifically amplifying DNA of tiger species , which could be applied to tiger species identification for doubtful samples or unable recognized products obtained in field investigation or forensic tests , mtDNA Cty b sequences of 5 tiger subspecies and 6 other cat species as well as 6 deer species were downloaded from Genbank , and compared by Wdnasis V2.5 software. Considering the essential doctrine of designing PCR primers , two regions in which all tigers share an unique hyplotype against other species were selected to design a pair of primers (Primer 1 , 2) to amplify a tiger Cty b fragment specifically. PCR tests were performed with DNAs extracted from muscle , visceral tissues , skins or hairs of Siberian tigers ( Panthera tigris altaica) , South-China tigers ( Panthera tigris amoyensis) and 6 other feline species and 6 non-feline species using this primer set. The results indicated that the primers are specific to tiger DNA and identification of tiger species by this means is accurate , sensitive , reproducible and reliable.

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The Phylogenetic Relationship among Goat , Sheep and Bharal Based on Mitochondrial Cytochrome b Gene Sequences

CAO Lirong WANG Xiaoming RAO Gang WAN Qiuhong FANG Shengguo

2004, 24(2):  109-114. 

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The blue sheep ( Pseudois nayaur) is a species currently listed as Endangered by IUCN and is registered in the state key protected wildlife list as a Category Ⅱspecies. It is widely distributed in most areas of Southwestern and Northwestern China. There has been no previous molecular examination of the phylogenetic origin of the blue sheep and our research results provide the first preliminary information and molecular perspectives on this problem. mtDNA Cyt b gene was used as genetic marker. Total DNA from three dried skin samples of bharal from Sichuan and Qinghai province in China were extracted and sequenced , and deposited in GenBank ( accession number : AF473606 , AF473607 , AF473608). These complete cyt b gene sequences (1 140 bp) , together with those sequences from goat ( Capra hircus) , ibex ( Capra ibex) , sheep ( Ovis aries) and argali ( Ovis ammon) in the family Bovidae which were obtained from GenBank , were aligned with the MegAlign software. By pairwise comparison , their Kimura 2-parameter distances and base substitutions were analyzed with MEGA software and the percentage differences among them were calculated with Dnastar software. According to two Cyt b molecular clocks , the time when different species began to evolve divergently was estimated. The phylogenetic trees constructed by two methods ( NJ and MP) supported almost the same topology.
Our molecular results showed that the mean sequence difference between blue sheep and the genus Capra (9.67 %) is significantly lower than that between blue sheep and the genus Ovis (11.78 %) . Sequence comparisons showed that the blue sheep is similar to the genus Capra. The phylogenetic analysis revealed that the blue sheep is more closely related to the monophylogeny of the genus Capra than to that of the genus Ovis. The approximate divergent time between bharal and the genus Capra is 3 - 6 Myr , whereas that between bharal and the genus Ovis is 6 - 8 Myr. The primitive-type karyotype is similarly maintained in Capra. A similar chromosomal complement of 2n = 60 is shared by the ibex and the goat. The diploid numbers of the chromosomes of the genus Ovis are 52 , 54 , 56 , 58 and that of blue sheep is 56. Therefore , we propose that blue sheep originated 3 - 6 million years ago when it split up from the genus Capra and developed separately. Further researches are needed in the future by being combined with the fossil records , the characteristics of morphology , behavior biology , and cytogenetics.

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Studies on the Karyotype and Chromosomal G2Banding of

ZOU Fangdong YUE Bisong ZHANG Yizheng CHEN San

2004, 24(2):  115-120. 

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A unique culture system has been developed to induce more peripheral blood lymphocytes of musk deer (Moschus berezovskii) into cell cycle by adding an equal quantity of phytohemagglutinin (PHA) and concanavalin A (ConA) , which stimulate G₀ phase lymphocytes to proliferate. Peripheral blood samples from 5 musk deers (3 ♂、2 ♀) of Institute of Scihuan Musk Deer Breeding was obtained from ear vein with heparin as an anticoagulant , and inoculated 0.3 - 0.4 ml of blood into 54 ml of RPMI 1640 medium containing 1 ml of bovine serum , 40μg of PHA and ConA , 0.05 ml of heparin (10 mg/ ml) and an appropriate antibiotics. The pH value of medium was adjusted to 7.0 - 7.21 The blood was cultured for 75 hrs and added colchicine to a final concentration of 0.01μg/ ml before the end of culturing. Preparation of chromosome was carried out with air-dry method and displayed G-bands with 0.025 % trypsin. It was found that the karyotype of M. berezovskii was 2N = 58 , and all of the chromosomes are telocentric ones. The results indicated that using late metaphases , in which all the chromosomes are linked only by centromeres , can determine whether the chromosome is telocentric or not. This study was the first time to use G-banding technique to analyze the chromosomes of M. moschiferus.

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Small Mammals Consuming Tree Seeds in Dujiangyan Forest

XIAO Zhishu ZHANG Zhibin

2004, 24(2):  121-124. 

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Through the live-trap-and-feed method (i.e.by placing three seeds each tree seeds and peanuts in live traps) , this study was conduct to investigate the species of small mammals , which potentially consume the six tree seeds : Quercus varialilis , Q. serrata , Castanopsis fargesii , Lithocarpus harlandii , Cyclobalanopsis glauca and Camellia oleifera in the forest of Dujiangyan Region , Sichuan Province , China. The results showed that : dominant rodent species , Leopoldamys edwardsi , Niviventer fulvescens , N. confucianus , Berylmys bowersi , Apodemus chevrieri and Rattus nitidus consumed all the seed species provided , and they also had some selectivity on these tree seeds. Leopoldamys edwardsi had a higher selective index for all tree seed species , while the other species had a strong selection on these seeds. Among these six tree seeds , all small mammal species preferred to consume Castanopsis fargesii seeds (with lower tannin) , but consume fewer Lithocarpus harlandii seeds (with hard seed coat) . This indicated that body size of small mammals , seed quality and the time to comsume a seed might be the main factors affecting seed selection by small mammals. This study also showed that the live-trap-and-feed method could inform the difference of seed selection by small mammals. But it had both advantages and limitations on seed selection by small mammals.

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Levels of Female Aggressiveness in Fluctuating Populations of Microtus ochrogaster and M. pennsylvanicus

Joyce E. Hofmann Lowell L. Getz

2004, 24(2):  125-131. 

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Levels of aggressiveness of females from free-living populations of Microtus ochrogaster and M. pennsylvanicus were compared during phases of the population cycle. M. ochrogaster displayed no significant difference in the proportion of aggressive females or in the means of five behavioral variables (approaches , threats , attacks , retreats , attacks/ approaches) among phases of the population cycle. None of these variables showed a significant linear regression against population density. Neither did the proportions of aggressive M. pennsylvanicus females differ among phases. The mean number of threats in this species was significantly higher during the low than the increase and decline phases and the mean number of attacks was significantly higher during the low than all other phases.The results of this study are similar to those obtained for males and do not support the polymorphic behavior hypothesis regarding generation of population cycles in arvicoline rodents.

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Seed-hoarding Behavior of Wild Apricot and Liaodong Oak by Small Rodents

LU Jiqi ZHANG Zhibin

2004, 24(2):  132-138. 

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In Donglingshan mountain (40°00′N , 115°30′E) , a mountainous area of Mentougou District of Beijing , China , we chose 30 seed releasing plots in a shrub community and released seeds of wild apricot ( Prunus armeniaca) and Liaodong oak(Quercus liaotungensis) , labeled with small pieces of coded tin-tags to investigate and compare the seeds of these two species eaten on the spot and survival dynamics and the distance , location and status of seed-transported. Furthermore , we identified rodent species by using comparison of gnawing marks on seeds of wild apricot made by different rodents. On 10th October 2002 , we released seeds of wild apricot and Liaodong oak , with an interval of 1 m between releasing places of them , on the ground surface in each plot , and checked the seed fate daily for 16 days. The recording items included number of seed-remained , transporting distance and micro-habitat and status of the seeds after transport. The micro-habitats were classified into four categories (Under shrub , US; Shrub edge , SE; Grass , G and Bare ground , BG). The statuses of those seeds being removed were defined as four types (Buried , B ; Surface , S; Eaten , E and Missing , M). SPSS for Windows (Version 10.0) was used in the analysis of data : survival analysis was used to analyze the remaining dynamics of the experimental seeds in the releasing place ; Kolmogorove-Smirnov test was used to determine the distribution of all data ; Mann-Whitney U was used to test the significant level of transporting distance between the removed seeds of wild apricot and Liaodong oak ; One-way ANOVA was used once the distribution pattern of data were normal. Chi-square test was used to determine if the two kinds of transported seed distributed randomly within four categories of microhabitats. The main results of this study are as follows : 1) Apodemus peninsulae and Sciurotamias davidianus are key rodent species acting on seeds of wild apricot and Liaodong oak in the study area ; 2) in releasing plots , the intensity of seeds of Liaodong oak were consumed is higher (54.38 %) than that of wild apricot (0.17 %) , the median survival time of released seed of wild apricot and Liaodong oak are 7.89 days and 7.73 days , respectively ; 3) the mean transport distance of seeds of wild apricot is longer than that of Liaodong oak ; 4) after transported by rodents , more seeds of wild apricot than seeds of Liaodong oak were buried within the micro2habitats of US and SE and 5) the cache size of seeds of wild apricot is 1 - 3 seeds with the majority of one seed , while the all caches of Liaodong oak has one seed only.

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Effects of Maternal Stress Induced by Predation Risk on Postnatal Body Mass Growth and Sex Ratio in Root Voles ( Microtus oeconomus

BIAN Jianghui WU Yan LIU Jike

2004, 24(2):  139-144. 

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Pattern of sex ratio variation and maternal investment in the literature are often inconsistent. This could be due to intraand inter-species variations in social system and the assumptions of the Trivers-Willard model not fulfilled in some studies. The present study was designed to test the main assumptions underlying Trivers-Willard model and whether the model could be explained the effects of maternal stress on growth and development of offspring and their sex ratio variation at weaning and adulthood in root voles. Pregnant mothers of root voles were exposed to a living steppe for 4 h daily. At birth and weaning , body mass of male and female offspring from stressed mothers were lighter than that of both offspring from unstressed mothers. Among them , female offspring could compensate for early growth disadvantage after weaning when they were allowed to unlimited access to food after weaning , whereas male offspring could maintain lower body mass into adulthood. Difference between treatment and control in birth sexual ratio per litter was not found. But litter sexual ratio in prenatal stressed offspring was strong skewed toward female at adulthood. Differences between males from stressed and unstressed mothers in mortality were also not found at weaning or adulthood. But mortality during from birth to adulthood in parental stressed male offspring was significantly increased compared with the control. We suggest that maternal stressful hormones secreted during gestation take part in sex-biased maternal investments effectively and directly and the consequences of the maternal investments accord with predication of Trivers2Willard mode.

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The Influence of Red Fox's Odor on the Reproduction of the Plateau Pika

WEI Wanhong CAO Yifan ZHANG Yanming YIN Baofa WANG Jinlong

2004, 24(2):  145-151. 

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The influence of the predation risk by using feces and urine of the red fox on the reproduction of the plateau pika (Ochotona curzoniae) was studied and the reproduction suppression hypothesis was verified in the field. The results showed that the body weight of the pika , which was the index to assess the reproduction investment of the pika , has no significant difference between treatment and control plots. The red fox's odor could not influence the reproduction investment of the pika. During the early period of their reproduction , both of male and female obviously increased their reproduction investment , but the investment of female was much higher than that of male during the late period of reproduction , because the male finished his mating activity and spent less time to care for the offspring than female. The reproductive success of the female , sexual ratio and resident rate for plateau pika have no difference between the treatment and control plots , but the reproductive success of male and the sexual ratio of the offspring for plateau pika were different between two plots. The pika could make decision and behaviorally decrease the predation risk. The results did not supported the reproduction suppression hypothesis and the predation risk did not suppress the reproduction of the plateau pika significantly.

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Energy Metabolism and Thermoregulation of the Desert Hamster (Phodopus roborovskii) in Hunshandake Desert of Inner Mongolia , China

ZHAN Xinmei WANGDehua

2004, 24(2):  152-159. 

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In order to understand the adaptive ecophysiological characteristics of desert hamsters ( Phodopus roborovskii) lived in Hunshandake desert of Inner Mongolia , we measured the energy metabolism and thermoregulation of adult hamsters. Metabolic rates were measured at a temperature range from 7 to 35℃using the closed-circuit respirometer , and the nonshivering thermogenesis was induced by subcutaneous injection of norepinephrine. The energy intakes were estimated with food trials. The body masses and body temperatures of animals were measured before and after each experiment. The thermal neutral zone (TNZ) of desert hamsters was 25 to 33 ℃, basal metabolic rate (BMR) was 2.61 ±0.04 (mlO₂/ g·h) , which is 152 % and 180 %of the values predicted by Hayssen &Lacy (1985) and McNab (1988) based on body mass , respectively. Mean body temperature of desert hamsters at the temperature range of 7 to 31 ℃was 35.7 ±0.1 ℃. The minimum thermal conductance within the temperature range of 7 to 25 ℃was 0.21 (mlO₂/ g·h·℃) , which is 98 % of that predicted by Bradley &Deavers (1980 ) . The maximum nonshivering thermogenesis of desert hamsters was 8.53 ±0.28 mlO₂/ g·h , which is 114 % of the value predicted by Heldmaier (1971) based on body mass , and the nonshivering thermogenesis scope (ratio of maximum NST to BMR) was 3.3. The energy intake ,digested energy , and metabolizable energy intake was 2.26 ±0.12 kJ / g·d , 2.18 ±0.13 kJ / g·d , and 2.13 ±0.12 kJ / g·d , respectively , while the digestibility and metabolizable energy efficiency was 97±0.2 %and 94 ±1.2 % , respectively1 Digestibility is higher in desert hamsters than other rodent species. No significant relationship between basal metabolic rate and digested energy was detected and , however , there was a significant relationship between minimum thermal conductance and energy intake ,digested energy , and metabolizable energy intake. These results suggest that high BMR and NST, wider TNZ, lower low critical temperature , relative lower body temperature , thermal conductance equal to the predicted value , and high digestibility can enable desert hamsters to live in an arid environment with high fluctuations in environment temperature and food resources.

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Neural Regulation of Pair Bond Formation in a Monogamous Rodent Species

Brandon J.Aragona J.Thomas Curtis LIU Yan WANG Zuoxin

2004, 24(2):  160-172. 

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The neurobiology of monogamous social organization can be studied by laboratory examination of social bonding. In this review , we discuss how the monogamous prairie vole (Microtus ochrogaster) has been used as a model system to provide tremendous insight into the neural regulation of pair bond formation. Neuroanatomical differences between monogamous and non-monogamous voles , as well as how neurochemical manipulations affect pair bond formation are reviewed. In addition , interactions among neurochemical systems that regulate pair bond formation and the extent of sexual dimorphism associated with pair bonding are discussed. Finally , we propose future directions for this line of research and explain why understanding the neural regulation of social bonding is important for human health.

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Reproductive Behavior Variations and Reproductive

LIU Xueqing ZHANG Zejun WEI Fuwen LI Ming LI Chun YANG Zhi HU Jinchu

2004, 24(2):  173-176. 

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In order to investigate the change of reproductive behaviors and understand reproductive strategies of both male and female red pandas , one-year behavioral observation was conducted through the focal sampling method in the Chengdu Research Base of Giant Panda Breeding from December 1999 to November 2000. Our results indicated that reproductive behaviors showed significant differences between the estrous and non-estrous seasons. Frequencies of the rubbing anogenital , sniffing and licking marking were much higher in estrus than in non-estrus. Bleating only appeared in the estrus and can be regarded as an estrous indicator.The result also demonstrated that both male and females applied different reproductive behavioral strategies. Frequencies of activity , rubbing anogenital , licking and sniffing marking , and bleat were much higher in the male than in the female. However , those of resting and investigating were much lower in the male than in the female. This indicated that the male was more active than the female during the estrus and might imply that the male acts mainly as an estrous message sender , and the female as a message receiver in the estrous season.

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The Sex Ratio of Plateau Pika's Population

WANG Jinlong WEI Wanhong ZHANG Yanming YIN Baofa CAO Yifan

2004, 24(2):  177-181. 

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The sex ratio ( ♂/ ♀) of plateau pika's ( Ochotona curzoniae) population was studied by re-captured method in the region of the Haibei Alpine Meadow Ecosystem Research Station , Northwest Institute of Plateau Biology , the Chinese Academy of Sciences from April of 2001 to August of 2002. The result showed that there was no significant difference from 1:1 in adult's sex ratio in whole breeding season , whereas the sex ratio of juvenile had some fluctuations among different age stages. The sex ratio of the second litter varied significantly from embryo to 60-day-old , but no difference at the first and the third litter. We concluded it was caused by conflict between maternal strategy and juvenile's strategy. No significant differences of sex ratio were found both in adult and juvenile between 2001 and 2002. The sex ratio of plateau pika before and after overwintering did not vary. In summary , we proposed that sex ratio of plateau pika's population was not influenced by exogenous factors , but some self-regulation mechanisms may be involved.

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Feces-based Detection of Yersinia enterocolitica in Intestine of Ailuropoda melanoleuca by PCR

YANG Shuiyun PEI Weijing SUN Feilong CHEN Xi WU Mingyu SHI Xiaoqiang WU Xiaomin REN Jianshe JIA Kangsheng

2004, 24(2):  182-184. 

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It is inevitable to develop noninvasive sampling methods to do studies on giant panda even diagnose the diseases since which is so endangered that it's impossible to carry out invasive sampling. A non-invasive sampling method to detect the intestinal pathogen , Yersinia enterocolitica in feces of pandas based designing PCR primers was established in this study. The main procedures are based on bacteria enrichment and cell lysis before binding the pathogen DNA to silica powder at high concentration of Kalium iodide and neutral pH conditions. Before PCR cycles , the binded DNA is washed with 80% ethanol and eluted with diluted EDTA buffer. Results showed that the silica-based feces DNA-purification method could remove the inhibitors of PCR so applicable to detect the target pathogen.