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07 February 2015, Volume 35 Issue 1

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 Evaluation of habitat for Asian elephants (Elephas maximus) in Xishuangbanna, Yunnan, China

LIN Liu, JIN Yanfei, YANG Hongpei, LUO Aidong, GUO Xianming, WANG Lifan, ZHANG Li

2015, 35(1):  1-13. 

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The Asian elephant (Elephas maximus)is listed under the first class of national protected species in China, and the majority of wild elephants live in Xishuangbanna,Yunnan Province. In this study,we applied Ecological Niche Factor Analysis (ENFA)with 3S technique to evaluate habitat quality to predict suitable habitat for Asian elephants in Xishuangbanna,using the distribution data of elephants collected between 2003 and 2008. Based on ENFA,the marginality value was 1. 375 and the tolerance value was 0. 478,which showed that the selection of environmental variables by the elephants was non-random,though they had certain tolerance and adaptability to the environment. The primary factor affecting habitat quality was the vegetation,specifically the bamboo forest,indicating that food is the most important factor for elephants in selecting suitable habitat. As the elephant distribution data in farmlands made up 45. 2% of all data points, we conducted ENFA using all distribution and non-farmland data respectively. When using all data points,the total area of
habitat,including optimal,suitable and marginal habitat,for elephants was 5 507. 9 km² and the farmlands accounted for35. 7% ,while using non-farmland data points,the total area of habitat decreased to 2 855. 4 km² and the farmlands accounted for 6. 3% . Therefore,the habitat predicted by non-farmland was the real suitable habitat for wild elephants in Xishuangbanna and is scattered in Mengyang,Mengla,Shangyong,Menglun, Naban-River reserves and their surrounding areas. Currently,the natural habitat of wild elephants is isolated by farmlands,and corridors should be established in selective areas to conserve the wild elephants more effectively.

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Abundance and distribution of the diet of the Sichuan snub-nosed monkey (Rhinopithecus roxellana) in the Shennongjia Nature Reserve in China

LI Jinyu, ZHANG Jing, TIE Jun, WU Changke, ZHANG Zhixiang

2015, 35(1):  14-28. 

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Primates living in temperate regions often suffer from dietary stress during periods of food scarcity．The Sichuan snub-nosed monkey (Rhinopithecus roxellana)lives in such a region． In the present study,we investigated vegetation and dietary plant species in habitats of R. roxellana at Qianjiaping in the Shennongjia Nature Reserve (SNR)in western Hubei Province,China,which is the eastern boundary of the range of the Sichuan snub-nosed monkey- We present data on food composition and abundance in relation to the monkey’s habitat choices. We chose critical and representative vegetation in
Qianjiaping,outlined 88 plots with an elevation gradient,and collected floristic data from 2010 to 2012- Based on dominant tree species,we divided the habitats into 12 types. Broadleaf forests contained more diet species for R. roxellana than conifer-broadleaf forests,and did habitats at lower elevation provided more dietary species than did those at higher elevation. Although R. roxellana are capable of eating many plant species,its preferred food plants were distributed unevenly among its habitats,and the quantity and composition of diet plants varied seasonally. We therefore suggest improving forest structure for the monkey by cultivating its preferred food plants,such as Euonymus spp. ,Malus spp. ,and Crataegus spp. ,or by controlled logging.

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Mating strategies of subordinate males in Tibetan macaques(Macaca thibetana) at Mt.Huangshan, China

LI Zhaopeng, LI Jinhua, XIA Dongpo, ZHU Yong, WANG Xi, ZHANG Dao

2015, 35(1):  29-39. 

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Abstract:In nonhuman primates with a linear dominance hierarchy,dominant males have priority of access to females in the social group,while,subordinate individuals have fewer mating opportunities． One of the fundamental questions is how to obtain mating opportunities and to increase the reproductive success for subordinate individuals． However,although this issue has been more or less ignored in previous studies,it can shed light on the mating system and mating strategy． In mating season (from September to December 2012),we used focal animal sampling,alloccurrence recording methods to collect behavioral data from one group wild Tibetan macaques (Macaca thibetana)at Huangshan,China (YA1)． We discuss the mating strategy of subordinate Tibetan monkeys． The results show that the frequency of copulations between subordinate males and females was higher when higher-ranking males were absent than when dominate males were present． Most mating events of subordinate males were more frequent,clandestine,and forced than those of higher-ranking males． Furthermore,higher-ranking males preferred to choose higher quality adult females,subordinate individuals had no preferred mating partners． During mating efforts,dominant males followed the females more frequently． However,subordinate males
obtained mating opportunity by grimacing and sexual chasing． Thus,we concluded that subordinate males formed the changeable and opportunistic mating strategy,most of their copulatory behaviors were forced mating.

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PAE coding system-based ethogram of Yangtze finless porpoise (Neophocaena phocaenoides asiaeorientalis) in a semi-natural environment

Chen Ran, Wei Yaolin, Wu Liang, Zheng Bangyou, Li Jinhu

2015, 35(1):  40-47. 

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The identification of species-typical behaviors and establishment of ethograms are fundamental for studying animal behavior and exploring the relationship between environment and behaviors. From August 2013 to April 2014, we used both coastal stationary and mobile tracking methods to observe 8 Yangtze finless porpoises (Neophocaena phocaenoides asiaeorientalis) living in a semi-natural environment located in Tongling Freshwater Dolphins National Nature Reserve. Using focal animal and ad libitum sampling methods, we recorded behaviors and coded them under the “posture-act-environment” (PAE) coding system. A total of 8 postures, 33 acts and 46 behaviors were recorded, and the relative frequency of each behavior related to gender and age was qualitatively described. The ethograms will enable further behavioral research and promote the protection of this animal.

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Preliminary analysis on taxonomic status of golden wild yak in Tibet

HOU Yunyun, ZHANG Yuguang, LU Hui, LIU Fang, LI Diqiang, FENG Jinchao

2015, 35(1):  48-54. 

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A population of 200 individuals of the golden wild yak inhabits the Chang Tang National Nature Reserve in Tibet. Golden wild yak is characterized by its unique golden hair compared to other black yaks. The genetic information for golden wild yak population is currently unknown due to its small population size,elusive behavior,and poor accessibility of their habitat. In this study,the taxonomic status of golden wild yak population has been preliminarily discussed based on two mitochondrial DNA fragments (i. e. ,Cyt b and D-loop)． The results show that the sequence characteristics,length, nucleotide composition of the two gene fragments of golden wild yak are similar to that of black wild yak and domestic yak.  Golden wild yak is the closest relative to other black wild yaks because the genetic distance between them is the smallest and they are clustered to a clade in a phylogeny tree of Bovine. However,the genetic distance between the golden wild yak and other black wild yaks is larger than the average genetic distance among the black wild yak individuals. Together with the morphological discrepancies,it is suggested that the golden wild yak is a subspecies or an important protection unit of wild yak. Due to the limited sample quantity in the present study,however,further evidence is still needed to corroborate our conclusions.

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Paternity identification of red panda using microsatellite genotyping

XIU Yunfang, LI Bichun,CHEN Yucun, ZHANG Yani, XU Suhui, El-Sayed Ahmed Kamel

2015, 35(1):  55-64. 

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The red panda (Ailurus fulgens) is an endangered species inhabiting a narrow region between the Himalaya Mountains and the Henduan Mountains． The studbook of captive red pandas in China has 968 registered individuals,of which 355 are living (as of 2013)but in this studbook,some pedigree information is unclear． In order to revise this studbook,microsatellite amplification was conducted on 41 captive red pandas with 19 pairs of red panda specific primers, and paternity relationship was analyzed. The paternity test results and species’original record information were analyzed using Pedigraph Software,and a genetic pedigree of Fuzhou captive red pandas was established．Our findings show that when the mother-child relationship was known,the combined paternity exclusion probability of the 19 loci was 0. 99999968．Genotype reliability testing of the 19 loci in six offspring with known parents revealed that the genotypes of 19 loci of offspring and parents completely comply with Mendelian Law,indicating that these loci can be used to ascertain the paternity of red pandas. The biological fathers of 15 offspring from the Fuzhou captive population were identified successfully. Paternity results showed that every litter has only a single father in spite of mating some female red pandas with multiple males in one breeding season. By genotype comparison,seven pairs of twins in the Fuzhou population were all found to be dizygotic twins and with the exception of studbook individuals #920 and #921,all of the other offspring from the Fuzhou population were sired by two males,#487 and #898,indicating different reproductive success rates．Our findings have practical significance for
strengthening the management of endangered animal populations and designing scientific breeding plans using modern breeding technologies.

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Composition and nutrient analysis of captive giant panda diet

YUAN Shibin, QU Yuanyuan, ZHANG Zejun, ZHAO Jingang, HOU Rong, WANG Hairui

2015, 35(1):  65-73. 

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In order to provide a reasonable feeding proposal for captive giant pandas in Chengdu Research Base of Giant Panda Breeding,we collected feeding data of 10 adult giant pandas (5 males and 5 females)during the past 4 years,observed foraging behaviors of 3 male adults for more than 1 year,and analysed nutrients and total flavonoids of food supplied. The results indicated that the giant pandas ingested about 10 main food bamboos and 3 bamboo shoots,and most preferred for Bashania faberi (the feeding duration was 232. 5 days),followed by Phyllostachys nidularia (the feeding duration was 134.5 days)and then by shoots of Qiongzhuea opienenss,Phyllostachys nidularia and Chimonobambus quadrangularis(the total feeding duration was 114. 7 days)． Judging from bamboos ingested and their seasonal foraging habits,giant pandas have developed relatively stable diet selection. The nutrient content of feeding bamboos has a certain regularity:the crude protein,crude fat,protein to energy ratio,calcium and total flavonoids concentrations of bamboo leaves were the
highest,but dry matter,crude fiber and phosphorus contents of branches and stems were the highest. We found that palatability,nutrient composition and content of total flavonoids of the main food bamboos are confirmed as the main factors influencing diet composition of captive giant panda at the research base．

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Response patterns of phytohemagglutinin-induced skin swelling at different reproductive phases in female striped hamster（Cricetulus barabensis）

ZHANG Zhiqiang, HUANG Shuli, ZHAO Zhijun

2015, 35(1):  74-79. 

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Phytohaemagglutinin (PHA)induced swelling response is an immunological parameter,which is widely used in the field of ecological immunology The swelling area of PHA post-injection is roughly equal to the strengths of the cell- mediated immune response and innate immunity as a comprehensive variation. To explore the response patterns of PHA-in-duced swelling during reproduction in small mammals,we measured the dynamic variations of skin tissue thickness at 0 h pre-injection,as well as 6 h,12 h and 24 h after PHA versus PBS injection at different reproductive phases (non-productive, pregnant, lactation,and post lactation phase)in striped hamster,Cricetulus barabensis. The results were as follows:(1)Hamsters showed two types of PHA response at different productive phases. One type response peaked 6 h after PHA injection for the non-reproductive,pregnant,and post lactation phases,then gradually declined at 12 h and 24 h. The other response did not show significant differences among different time points for the lactation phase. (2)Maximum PHA response for the lactation phase showed a reduced tendency,but no significant differences were observed among different reproductive phases. Our results indicate the PHA response pattern for the lactation phase is different from that of other reproductive phases,and maximum PHA response is delayed,which may be harmful to maternal survival in a short term,while it is beneficial to promote reproductive process and accelerate offspring’s growth and development.

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Distribution and density of sweat glands in Mongolian gerbils（Meriones unguiculatus）and Brandt’s voles ( Lasiopodomys brandtii）

XU Mengmeng，WANG Dehua

2015, 35(1):  80-86. 

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Mammals can sweat to regulate body temperature when exposed to high ambient tenperature,but it has been stated that rodents do not sweat,and there is little information on sweat glands in wild rodents. By using hematoxylin-eosin staining techniques,we examined the skin morphology,sweat gland distribution and density in Mongolian gerbils (Meriones unguiculatus)and Brandt’s voles (Lasiopodomys brandtii)． We did not find sweat glands on the skin of head,angulus oris,stethidium,armpit,abdomen,dorsum,and hindleg,but eccrine sweat glands on hairless palmar and hairy plantar skin were found in both Mongolian gerbils and Brandt’s voles. Density of sweat glands of  fore-paws (2. 40 ±0. 49 // mm² )was higher than that on hindpaws (0.15 ±0. 03 // mm² )in Mongolian gerbils,while there was no significant difference between fore-paws (0. 37 ±0. 05 // mm² )and hind-paws (0.21 ± 0.08 // mm² ) in Brandt’s voles. It seems that sweat gland density might be related to the hair condition of paws,density is low when the paw skin is hairy. We predict that the function of sweat gland in Mongolian gerbils and Brandt’s voles may help to increase heat dissipation and increase friction for locomotion.

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Suppression effects of quinestrol on the reproduction of Brandt's voles(Lasiopodomys brandtii)

WANG Taotao, GUO Yongwang, WANG Deng, HAI Shuzhen

2015, 35(1):  87-94. 

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It is critical to confirm the optimum dose and the active duration of the sterilization by quinestrol on rodents for some studies and practices. We fed female and male Brandt’s voles 5 doses of 0. 5 mg// kg,1.0 mg// kg,2. 0 mg// kg, 3. 0 mg// kg,or 6. 0 mg// kg quiestrol for 7 consecutive days．We then assessed their anti-fertility effects on the voles by analyzing reproduction indices including birth date,reproductive rate and reproductive capacity. Results showed that quinestrol delayed the breeding of Brandt’s voles in the lab,but the effect was not significant due to the small sample size. In 90 days,some doses of quinestrol decreased the reproductive rate and average litter size of female and male,however,the inhibitory effect still was not significant,again possibly due to small sample size;some doses of quinestrol decreased markedly the reproductive capacity of Brandt’s voles. The suppression effects were not significantly correlated with the doses of quinestrol. The optimum doses of quinestrol causing sterility were below 2. 0 mg// kg for male and 1. 0 mg// kg for female. The dose applyed for Brandt’s vole should be below 2. 0 mg// kg,which can reduce total litter size of Brandt’s vole obviously within 90 days.

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The effect of cold stress on the HSP70 mRNA expression in mammary epithelial cells of yak

JING Muxian, CUI Yan, YANG Kun, WEN Zexing, LIU Penggang, SHEN Qingfeng,YU Sijiu

2015, 35(1):  95-101. 

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The present study was carried out to investigate the effect of cold stress on the expression of Heat shock protein 70 (HSP70)in mammary epithelial cells of the yak. The changes of HSP70 mRNA expression in the condition of cold stimulation of 10℃ were determined by use of realtime fluorescent quantitative PCR technology.The results showed that no significant changes (P >0. 05)of HSP70 mRNA expression would be found if the mammary epithelial cells were treated only in the condition of 10℃ for 2 h,4 h,6 h and 8 h,respectively. A significant increase of HSP70 mRNA expression would be found and reached a peak value (P <0.01)at the 6th hour of cold treatment if the mammary epithelial cells were first treated in the condition of 10℃ for 2 h,4 h,6 h and 8 h,respectively,and then reheated for 4h. A significant increase of HSP70 mRNA expression would also be found and reach a peak value (P <0. 01)at the 4th hour of rewarming if the mammary epithelial cells were first treated for 4 h in the condition of 10℃,and then reheated for 2 h,4 h,6 h and 8 h,respectively. It is concluded from this study that an increase of HSP70 mRNA expression does not occur in the process of cold treatment,but in the process of rewarming; meanwhile the increase of HSP70 mRNA expression is to some extentrelated with the time duration.

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Development of a SYBR Green I real-time quantitative PCR assay for the detection of Mink enteritis virus

YU Yongle, ZHANG Chuanmei, YANG Haiyan, YANG Ruimei, ZHANG Hongliang, SHAN Hu

2015, 35(1):  102-109. 

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A quantitative PCR assay was developed for specific and sensitive detection of mink enteritis virus. The primer pair targeting a 146 bp fragment of the conserved VP2 gene was selected based on alignment of viral sequences available at GenBank. The analytic sensibility of this assay was 34 copies of plasmid DNA,and it had high repeatability with CV <2% ． The assay was evaluated by testing 80 fecal samples collected from suspicious cases. The results showed that 30 samples were found positive for the virus infection,which was higher than that of routine PCR and electron microscopy. Therefore the new assay provides a useful method for the rapid diagnosis of mink enteritis and the prevention and control of the disease.

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The application of extracting DNA from noninvasive samples in feline species identification

ZHOU Xuwei, SONG Jingliang, WANG Fuyou, XIA Wenjun, WU Liji, ZHANG Zhenhuan, BAO Weidong

2015, 35(1):  110-118. 

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Field surveys utilizing indirect indicators are often adopted in population ecology studies of wild cats in China．However,research using molecular biology techniques are still relatively rare． Confirmation of molecular techniques as an informative method for wide use in population analysis is required． Therefore,in this paper,molecular markers(mtDNA,sex gene and microsatellites)were used for determining applicability in conservation genetic research by comparing fecal samples from 4 feline species． Nine of 11 fecal samples suspected for Siberian tiger were identified as Panthera tigris altaica and
16 of 30 fecal samples suspected as Eurasian lynx were confirmed as Lynx lynx． Additionally five tigers (four males and one female)and five Eurasian lynx (three males and two females)were identified． Genetic diversity analysis indicated that the average number of allele of microsatellite loci of the Siberian tiger was 2. 64;the average observed heterozygosity was 0. 489 and the average polymorphism information content was 0. 301,while these values for Eurasian lynx were 2. 88,0. 575,and 0. 428,respectively,indicating a moderate level of genetic diversity for the two species Analysis of Eurasian lynx fecal samples preserved in pure ethanol were as follows:species identification rate of 69% ;sex identification rate of 100% ;microsatellite amplification rate 85%. By comparison analysis of air dried samples were 41%,71%,and 25% for species identification,sex identification and microsatellite amplification, respectively． The species identification rate for fecal samples preserved for one year was 80%,the sex identification rate was 75% and the microsatellite amplification rate was 28% ． Our results provide a reliable method for high success in DNA extraction,from noninvasive fecal samples,for identifying wild cats and determining population genetic structure．