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02 March 2010, Volume 30 Issue 1

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Habitat selection in dry season of Asian elephant (Elephas maximus) and conservation strategies in Nangunhe National Nature Reserve,Yunnan, China

FENG Limin1 ,WANG Zhisheng2 ,LIN Liu,YANG Shaobing,ZHOU Bin,LI Chunhua,XIONG Youming,ZHANG Li

2010, 30(1):  1-10. 

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This work studied habitat selection of Asian elephants (Elephas maximus) in the dry season at Nangunhe National
Nature Reserve,Yunnan of China. Village investigation and line-transect samples were used during the field survey. Waypoints were made about 1 km apart along sampling lines with 195 km in total. Data were collected on vegetation type,vegetation abundance,geography parameters (aspect,slope,location and elevation),population status and frequency of elephant sightings. A total of 54 elephant trace points were tracked during the survey in the dry season of 2005. 3S (GIS,GPS, RS) technology was used to analyze the data obtained from field survey and satellite images. Vanderploge and Scavia’s selectivity index were used to assess Asian elephants’selection for different habitat parameters. The results indicated that elephants preferred habitat where elevation was less than 1000 meters,slope less than 20°, located in a valley and oriented north, southeast and south. Elephant-preferred vegetation included bamboo-broad-leaved mixed forest and shrub. Asian elephants showed a preference for denser coverage shrub layer and arbor layer with lower height,dense canopy and small DBH. Based on satellite images,we analyzed the forest dynamics from 1988 to 2007 and found heavy loss of suitable habitat which seriously
threatened the survival of Asian elephant in Nangunhe National Nature Reserve. Preventing habitat loss and degradation, and strengthening the control of poaching would be the key tasks for the conservation of elephants in Nangunhe.

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Habitat suitability assessment of Gazella subgutturosa in Kalamaili Mountain Nature Reserve

LI Ying,XU Wenxuan,YANG Weikang,QIAO Jianfang,LIU Wei,XIA Canjun

2010, 30(1):  11-20. 

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We selected a representative region of about 1447 km² for an assessment area in the Kalamaili Mountain Nature
Reserve. Based on the data obtained from field surveys of Goitred gazelle (Gazella subgutturosa) in Kalamaili Mountain Nature Reserve since 2005, we used the habitat evaluation model and Geographic Information System (GIS) to assessed suitability of the habitat for Goitred gazelle. Vegetation types,slope,aspect,and human activities were selected as factors for habitat suitability assessment. The results showed that the area of actual habitats was 1146. 9 km² ,1137.1 km² ,991.6 km² and 499.8 km² respectively during four seasons;accounting for 79.3% ,78.6% ,68.5% and 34.5% of the total assessment area. The area of the suitable actual habitats for Gazella subgutturosa was 304.1 km² ,599.4 km² , 303.6 km² and 56. 2 km² respectively in the four seasons;accounting for 21. 0% ,41.4% ,21.0% and 3. 9% of the total assessment area. Some potential habitat has been transformed into unsuitable habitat. The habitat lost is 3.9% ,3.1% ,4.3% and 48.5% during the four seasons. Among human interferences,residential areas,a national highway and mining affect the habitat,but have little impact. In winter,suitable habitat for gazelle was occupied by livestock,which leads to a decline in the quality of habitat in the large area. Based on evaluation results,this paper also put forward measures to protect them.

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Activity rhythms of captive white-headed langur in spring

PENG Yanyan,CHEN Min,WANG Aishan,YAO Jianzhuang,ZHANG Endi

2010, 30(1):  21-27. 

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Five captive white-headed langurs (Trachypithecus leucocephalus) were observed day and night from March to
May, 2008 in Shanghai Zoological Park using an instantaneous scan sampling method. Results showed that the langurs activity began at around 06∶ 00 and ended at 18∶00 or 18∶ 30 in spring. During the daytime, resting, feeding and moving were the main behaviors of the langurs. The mean frequencies were 13.79, 4.75 and 2.18, respectively. During the nighttime, the main behaviors were resting,moving and scratching The mean frequencies were 22.13, 0.43 and 0.26, respectively. Activity rhythm of moving,grooming,playing and other behaviors differed significantly among the individuals. No grooming and playing behaviors happened at night. The langurs had four small movement peaks at 21∶ 00,00∶30, 03∶ 00 and 04∶ 30, and also had scratching peaks at 22∶30, 01∶ 00, 02∶ 03∶30, 04∶ 30 and 05∶ 30 from which we inferred that the langurs remained awake and alert during the long night. Moving behavior differed significantly between male and female langurs during the day and night.

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Population size and habitat of wild boar (Sus scrofa) in the eastern Wanda Mountains

ZHOU Shaochun,ZHANG Minghai,SUN Haiyi,YIN Yuanxin,HUANG Haijiao,YU Hongwei,LU Xiangdong,GE Dongning,TIAN Jialong

2010, 30(1):  28-34. 

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We estimated population size and performed a preliminary analysis of habitat selection of wild boar using line
transects in Eastern Wanda Mountains,Heilongjiang Province,China from November 18, 2008 to March 20, 2009. We randomly set 40 sampling sites with 200 transect lines to collect field data on wild boars and their habitat at thirteen forestry farms operated by the Dongfenghong Forest Bureau. We estimated population density at 0.175 individuals/ km² and abundance at 546 - 680 individuals. They were distributed in an elevation range of 300 to 800 m. The distribution areas of wild boars were mainly concentrated in Hekou, Qiyuan, Qingshan, Wulindong, Dumuhe, Haiyinshan, and Donglin forest farms. The population sizes and densities of wild boars were 0.3720 individuals / km² and a total of 1 302 individuals in 1989,0.3423 individuals/ km² and a total of 1198 individuals in 1989. We found marginal evidence that the population size decreased slowly from 1989 to 2002,then decreased rapidly from 2002 to 2008. On the other hand,the percentage of wild boar tracks lest in snow in various forest stands shown that wild boars choose habitats of middle slope position a sunny aspect,slope angle less than 5°, 25 to 50 percent shelter and canopy density,vegetation coverage more than 30 percentage. Wild boars prefer broadleaf forest and shrub. Four threatening factors,including illegal hunting to wild boar,forest harvesting,nut collection,and predation by Amur tigers contributed to declining population size and habitat degradation.

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Influence of burrowing activity of plateau pikas (Ochotona curzoniae) on nitrogen in soils

LIU Wei,XU Qingmin,WANG Xi,ZHAO Jianzhong,ZHOU Li

2010, 30(1):  35-44. 

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We analyzed the effects of burrowing activity of plateau pikas on inorganic nitrogen in soils by measuring the total
nitrogen content,N-NH₄⁺ content,and N-NO₃^- content in different sorts of mounds soil and in soil 0 - 10 cm below the mounds,and estimated increased sizes of inorganic nitrogen by mean mounds density rate. The results showed that total nitrogen contents were not changed in different sorts of mounds soil,and the trend for NH₄ ⁺
nitrogen content,NO₃^- nitrogen content,inorganic content were:in new mounds > in two-year mounds > in multiyear mounds > in control soil. The one way ANOVA results showed that NH₄⁺ nitrogen content was not changed in different sorts of mounds soil,but NO₃^- nitrogen content in new mounds soil and in two-year-old mounds soil was greater than it was in multiyear mounds soil and in control soil in May (F = 14.55, P < 0. 001,df =3,12),and inorganic content in new mounds was greater than it in control soils in May and September (F_May =14.55, P <0.001,df = 3,12; F_Sept^.= 3. 92, P < 0.05,df =3,12). N-NH₄⁺ content reduced gradually from May to August,and increased in September,N-NO₃^- content and inorganic nitrogen content showed the trends of “high-low-high-low-high”from May to September. The one way ANOVA results showed significant differences between months. N-NH₄⁺content, N-NO₃^- content and inorganic nitrogen content in soils 0 - 10 cm under different sorts mounds showed the same change as in mounds soils among months and treatments,but N-NO₃^-content and inorganic content were greater in soils 0 - 10 cm under new mounds soil than it was in control soil (P < 0. 05) in the same month respectively,
and N-NH₄⁺content,N-NO₃^- content and inorganic nitrogen content showed significant differences between different months of the magnitude of the changes in N-NH₄⁺ content,N-NO₃^- content and inorganic nitrogen content were 162.6 mg/ kg, 355.1 mg / kg and 497.7 mg / kg respectively in new mounds soil.

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Growth and thermogenesis in pups of Apodemus chevrieri

LIU Pengfei, ZHANG Lin, WANG Rui, ZHU Wanlong, LIU Chunyan, MENG Lihua, WANG Zhengkun

2010, 30(1):  45-50. 

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Using 56 days of postnatal data from Apodemus chevrieri, this paper analyzed tentatively its growth rules. Electronic
scales and Open-Circuit Respirometry were used to measure body mass, resting metabolic rate,non-shivering thermogenesis, and evaporative water loss. A. chevrieri weight growth could be divided into rapid-growing and slow-growing situations based on the inflection point of a logisitic growth curve. Body temperature of pups increased with age,and approached the adult level at 35 days of age. Resting metabolic rate (RMR)and Nonshivering thermogenesis (NST) increrased with age at early time points and then showed allometry with body mass. RMR reached the adult level at 49 days of age. Onset of NST occurred during the 6th day after birth and its evaporative water loss (EWL) was higher during lactation than during the weaning period. A. chevrieri is an altricial mammal. The short gestation time,larger litter size,and longer lactation term of A. chevrieri are consistent with the relatively abundant food supply in its habitat.

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Maintenance of polymorphism of Chinese muntjac (Muntiacus reevesi) Mhc-DRB gene

YU Jian,LIANG Wenming,ZHU Feihu,XU Si,WU Hailong

2010, 30(1):  51-57. 

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A pair of primers (LA31 and LA32) specific to BoLA-DRB3 were used to amplify genomic DNA of the Chinese
muntjac (Muntiacus reevesi). Twenty distinct DRB exon 2 alleles were obtained from 12 individuals through polymerase chain reaction (PCR),single strand conformation polymorphism (SSCP),and sequencing. At least two DRB loci could be discerned taking into account the fact that three to four alleles were identified from six of 12 individuals analyzed in this study. No insertion,deletion or stop codons within the nucleotide sequences were detected. Multiple sequence alignments demonstrated that the Chinese muntjac DRB alleles showed high similarity to that of the expressed BoLA-DRB3 locus. The amino acid mutation sites were mainly present in the putative peptide binding region (PBR). These results evidenced that the loci isolated in this study were probably expressed and functioning. The frequency of nonsynonymous substitutions (d_N ) was significantly greater than that of synonymous substitutions (d_S )(P < 0.01) in the peptide binding region (PBR) suggesting that the DRB gene of the Chinese muntjac had historically undergone positive selection. This inference was further confirmed through likelihood ratio test (LRT)from model M7 and M8 of program CODEML. Six robust positive selection amino acid sites (position 11,37,61,67,71 and 86)were detected by a Bayesian method,of which 5 sites located in PBR. These results provide evidence that positive selection is one of the main mechanisms in maintaining DRB gene polymorphism of the Chinese muntjac. A neighbor-join (NJ) tree was constructed based on DRB gene exon 2 sequences of several deer. In the NJ tree,the Chinese muntjac DRB alleles showed a mosaic pattern with that of other deer, implying a trans-species evolution pattern in maintaining DRB allele polymorphism. Moreover,two DRB alleles of Chinese muntjac (Mure-DRB1 and Mure-DRB11) and two DRB alleles of red deer (Ceel-DRB34 and Ceel-DRB46)were closely clustered with DRB3 alleles of cattle,suggesting some DRB alleles of Chinese muntjac and red deer might share an ancient lineage.

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Population genetic diversity of roe deer (Capreolus pygargus) in Northeastern China

ZHANG Minghai,LIU Yanhua,JIA Jingbo

2010, 30(1):  58-64. 

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Roe deer (Capreolus pygargus) are important economic animals in China and are one of the major sources of
food for the Amur tiger. To design effective conservation strategies for roe deer, we investigated the genetic diversity and gene flow in eight roe deer populations by analyzing 463 base pairs of the mitochondrial DNA (mtDNA) control region in 88 individual deer sampled from Yinggchun, Dongfanghong, Shuangfeng, Taipinggou, Xinqing, Zhanhe, Shengshan and Tuqiang (Table 1). Fifty-nine variable sites and thirty haplotypes were identified. The roe deer exhibited high mtDNA diversity with haplotype diversity (H = 0.967 ±0.007)and nucleotide diversity (π =2.641 ± 0.02595). Neither the estimate of Tajima’s D nor that of Fu and Li′s D deviated significantly from the neutral selection hypothesis (P > 0.1) for all eight populations of deer,showing no evidence of strong selective sweeps or balancing selection (Table 2). Based on the analysis of thirty haplotypes,the roe deer can be divided into three subpopulations (Figure 1). Although there appears to be gene flow among the three subpopulations,these data indicate that the population of deer from Tuqiang is genetically distinct from the other population and is worthy of specific conservation efforts. We recommend that a breeding program be established for the Tuqiang population in the Great Khingan Mountains.

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cDNA cloning and sequence analysis of growth hormone receptor (GHR) from Arctic fox

DU Zhiheng,NING Fangyong,YANG Chunshan,BAI Xiujuan

2010, 30(1):  65-73. 

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In the current study,the gene encoding growth hormone receptor (GHR)was cloned by RT - PCR from Arctic fox (Alopex lagopus)according to dog GHR gene (AF133835). Sequence analysis revealed that the open reading frame
(ORF) of Arctic fox GHR is 1917 bp,which encodes a 638 - amino acid precursor containing a signal peptide of 18 amino acids and a mature peptide of 620 Amino acids. Arctic fox GHR gene has highest similarity (98% ) with dog GHR. The molecular phylogenetic tree was constructed by the NJ method based on GHR gene sequences. The result showed the Arctic fox and the dog were clustered into one branch,this result is consistent with traditional classification. In addition, Arctic fox GHR has its own characterization in its amino acid sequence compared to other mammal GHR sequences. For example, its amino acid residues at position 45 and 451 are A and E, respectively,while for others species the corresponding amino acid residues are T (K for rat )and A (V for cattle and sheep and T for mouse).

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Genetic cloning and expression of neuroglobin in high altitude hypoxic ad- aptation species——Plateau pika (Ochotona curzoniae)

BAI Zhenzhong,HAN Shufen,YANG Yingzhong,CAO Yue,MA Lan,LIU Shou,GE Rili

2010, 30(1):  74-78. 

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For identification of the neuroglobin genes coding sequences molecular cloning and examination of the tissues expression spectrums and showing the hypoxic adaptations mechanisms in Plateau pika (Ochotona curzoniae). Extracting the total RNA,cloning neuroglobin coding sequences cDNA with reverse-transcription RT-PCR,capturing and confirming the certain sequences with DNA sequencing. Examination the tissues mRNA expressions spectrums of neuroglobin with in suit hybridization ISH technology. In addition,semi-quantitative RT- PCR and western-blot technologies were being used to analysis of the relative neuroglobin mRNA and protein expression amounts in various tissues with Plateau pika. Cloning and sequencing results confirmed it was the neruoglobin gene coding sequence of Plateau pika with Blast analysis;and the ISH results exhibited the neuroglobin gene expressions spectrums indicate widely distributed with higher amount in brain tissues of Plateau pika. Additionally,neuroglogin mRNA also expressed in the other tissues such as testis and adrenal glands besides brain tissues. We can speculate that NGB might play an important role in the adaptations of the Plateau pika under the high altitude environment. Furthermore,native Tibetan species neuroglobin genes will also present fundamental evidence to investigate high altitude adaptable related genetics study in the future.

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Molecular cloning and eukaryotic expression of Canine distemper virus receptor gene SLAM of fox,raccoon dog and mink

ZHAO Jianjun,ZHANG Hailing,GAO Han,ZHAO Chunfei,CHAI Xiuli,CHEN Tao,YAN Xijun,WU Wei

2010, 30(1):  79-86. 

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Signaling lymphocyte activation molecule (SLAM)is one of the cellular receptors for Canine distemper virus (CDV) infection. In this study,we first cloned and sequenced the SLAM genes from peripheral blood lymphocytes of fox (Vulpes vulpes),raccoon dog (Nyctereutes procyonoides)and mink (Mustela lutreola) respectively. Identical with the dog (Canine) SLAM,SLAM cDNAs both of fox and raccoon dog were 1 029 bp length encoding 342 amino acids. The nucleotide sequence homologies of SLAMs among fox,raccoon dog and dog were highly homologous (>98.6% ). The SLAM cDNA of mink was 1 020 bp in length encoding a polypeptide of 339 amino acids and had high homology with spotted seal (Phoca largha) SLAM (91.4% at nt level,87.9% at aa level). Phylogenetic analysis based on the nucleotide sequences of SLAM genes of different species showed the natural hosts including dog, fox, raccoon dog, mink and seal for CDV formed a monophyletic cluster. In addition to the 26 amino acid residue predicted signal peptide of SLAMs,there were 8 amino acid residues in the V region of SLAM thought to be involved in determining host-virus specificity were completely conserved,irrespective of the species of CDV infection.We also established three eukaryotic recombinant plasmids using an pIRES2 - EGFP vector which expressed the SLAM cDNAs of fox,raccoon dog and mink in transient transfection CRFK cells. One day after inoculation with CDV virulent strain,all the SLAM - expressing CRFK cells produced extensive cytopathic effects (CPE). Our results suggest that CDV use SLAMs of fox,raccoon dog and mink as a cellular receptor.

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Nutritional requirement of captive non-human primates

ZHANG Peng,WATANABE Kunio

2010, 30(1):  87-98. 

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Nutritional requirements of captive non-human primates have increasingly become a topic of focus for international and Chinese communities. Introduction of nutritional studies on non-human primates to China,the biggest breeding and exporting country for non-human primates in the world,will help to improve the standard of raising the animals,the feeding efficiency of ingredients,and industry development. This article reviews recent research progress on the nutritional requirements of captive non-human primates and explains the recommended levels by both Chinese (GB 14924.8 - 2001) and US (NRC 2003)governments. It further introduces functions of each nutrient composition including the clinical and pathologic signs of deficiency and treatment for the deficiency. The purpose of this review paper is to enrich the relevant scientific information and to promote understanding of the nutritional needs of captive non-human primates.

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Cloning and sequence analysis of bitter taste receptor T2R2 gene in giant panda

XU Huailiang,YAO Yongfang,ZHU Qing,CHENG Anchun

2010, 30(1):  99-104. 

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Based on bitter taste receptor T2R2 gene sequence of domesticated dog (AB249685),one pair of primers were designed and used to amplify an approximately 1.1 kb DNA fragment from genomic DNA sample of giant panda by using PCR. The PCR products were ligated into the pMD-18T vector,and then transformed into competent cells of E.coli DH5α. The identified positive clone was sequenced. The result showed that the T2R2 gene of giant panda was 1 008 bp in length, and contained complete exon,and 915 bp,encoding 304 amino acid residues. The pI of the protein is 9.56 and its molecular weight is 34.90 kDa. The prediction of topological structure for the protein indicated that it contained 11 potential functional sites (three N-glycosylation sites,three potential protein kinase C phosphorylation sites,two N-myristoylation sites, one Casein kinase II phosphorylation site,one cAMP-and cGMP-dependent protein kinase phosphorylation site and one Nuclear Localization Signal site) with five sites observed only in giant panda,and the protein comprised seven transmembrane helix regions,and four extracellular regions and four intracellular regions. The T2R2 was a hydrophobic protein with less hydrophilic components which mostly were located on the intracellular regions. Alignment analysis revealed that the homology of T2R2 gene nucleotide sequence of giant panda with that of dog,cat,cattle,horse,chimpanzee and mouse is 92.65%, 91.12% , 85.64% , 86.73% , 85.20% , 72.59% , respectively, and the homologies of amino acid sequence is 86.73%, 85.20%, 74.67% , 78.62% , 75.66% , 60.53%, respectively. On the whole,the giant panda T2R2 gene was high evolutionarily conserved,but its protein presented more abundant functional sites than did those of other species. However,
the correlation between the characteristics of T2R2 gene and giant panda’s special diet needs to be further studied. The obtained sequences were submitted to GenBank,with accession NO. FJ812726.

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A new method for detecting exon 2 alleles of DRB genes in sika deer

LI Bo,XU Yanchun,MA Jianzhang

2010, 30(1):  105-109. 

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A new method was found which integrated Motif specific-PCR,SSCP and direct sequencing to identify the exon 2 of alleles of DRB genes in sika deer. Using this method,the detected 15 sequences which included no artificial sequences from mutation or recombination in the process all met the criterion to distinguish a new allele. The criterion is that there have to be identical sequences identified from at least two different individuals by PCR and direct sequencing. Compared with PCR-SSCP-cloning sequencing previously,this method can efficiently remove artificial sequences. Combined with the above criterion,this method provided a new approach for investigating DRB gene polymorphism of sika deer and other deer species.

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Seasonal variation of pelage characteristics in Siberian weasel (Mustela sibirica) of Xiaoxing’anling area,Heilongjiang,China

HUA Yan,ZHANG Wei,XU Yanchun

2010, 30(1):  110-114. 

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Coat characteristics of seasonal molting mammals reveal significant seasonal variation as an adaptive strategy to cope with seasonal climate changes. However,the adaptive significance of such morphological variation has not yet been addressed. We analyzed seasonal variation of microscopic indices of hair and skin of adult Siberian weasels (Mustela sibirica manchurica Brass) from the Tonghe forest area of the Xiaoxing’anling Mountains,Heilongjiang. Skins from 8 males and 8 females were collected from summer (July to September),and an additional 8 male and 8 females skins were collected from winter (November to December )(i. e. ,n = 32). Morphological indexes included length and width of guard hair, cuticular scale patterns of guard hair,external and cross-section form of guard hair,and medullary characteristics. We found significant differences between winter and summer coat hair density,hair length,and proportion of medulla-absent part of guard hair. We discuss the adaptive mechanism of this seasonal variation.

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New record of Formosan lesser horseshoe bat (Rhinolophus monoceros, Andersen 1905 Rhinolophidae,Chiroptera)in Mainland of China

ZHOU Jiang,YANG Tianyou

2010, 30(1):  115-118. 

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During a survey of bat species diversity from July 2008 to February 2009 in Songtao county of Guizhou Province, one specimen of Rhinolophus was collected from Guizhou. It was identified as a Formosan lesser horseshoe bat,based on the following features:small body size,forearm length of 36.3 mm,the greatest length of skull was 15.16 mm,the cells of lancet were shorter and appeared triangular and showed an acute angle,and forward bending,the lancet was shorter and approximately triangular and had a narrow top,the anterior median swellings of the skull were higher and more vertical,the rear of the sagittal crest was thin,the upper molars were shorter and the length of C¹ - M³ was 5.36 mm,the width of M³ -M³ was 5.24 mm. Rhinolophus monoceros,which is new record in Guizhou and the first found on the Chinese mainland, supports Simmons’prediction that Rhinolophus monoceros may occur in the south of China.