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Table of Content

    08 March 2009, Volume 29 Issue 2
    Foraging ecology of feral goats on the Isle of Rum,NW Scotland
    SHI Jianbin,Robin I MDunbar
    2009, 29(2):  116-124. 
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    We studied foraging ecology of feral goats (Capra hircus) on the Isle of Rum,Scotland,from June to November 2000. Grazing bout length varied greatly from 1 min to 460 min with mean duration being 103.1 ±15.0 (SD)min. Adult males and females differed marginally significantly in feeding bout length with females having relatively longer feeding bouts (P = 0.077). The average bite rate for feral goats was 46.3 ± 0.6 bites/ min with significant variations between sexes (P = 0.023)and among months (P <0.001). Adult males had faster bite rates during pre- (June-July)and post-rutting (October-November)periods than during rutting period (August-September)(P <0. 008),but the bite rate of adult females during pre-rutting period was not different from that during rutting period (P = 0.327). Adult males also spent significantly less daytime feeding during rutting period. The differences in bite rates and feeding time between the two sexes might result in sexual differences in forage intake by feral goats:females had relatively stable intake,but males fluctuated greatly in their forage intake. The estimated forage intake decreased from June to November (although a slight increase from September to November),which means that feral goats on Rum may be in negative energy balance during autumn / winter period when the quality of forage is lower and the weather is beginning to deteriorate. There was no significant difference in step rate while feeding between sexes or among months.
    Winter bed-site selection by argali Ovis ammon sairensis,O. a. darwini in Mt. Kekesen and Mt. Kalamaili in southern Altai Mountains
    CHU Hongjun,JIANG Zhigang,QI Yingjie,TAO Yongshan,LI Bin
    2009, 29(2):  125-132. 
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    Winter bedding sites used by argali Ovis ammon sairensis,O. a. darwini were studied by using line-transect
    methods on Mt. Kekesen and Mt. Kalamaili,in the southern desert of Altai Mountains,Xinjiang Ugyur Autonomous Region, China from November of 2007 to January of 2008. One hundred bedding sites used by argali and 100 control plots were 1ocated and measured during a 30 transect surveys crossing the entire study area on Mt. Kekesen,while 125 sites used by argali and 125 control plots were 1ocated and measured during 45 transects surveys crossing the entire study area on Mt. Kalamaili. Argali at both sites preferred slopes as bed sites,and avoided sleeping in valleys and on plains;argali showed a preference for slopes which were half exposed to the sun and avoided shady slopes;argali showed a preference for the upper slope and the middle slope. Comparing bedding sites with control plots on the two mountains bedding sites of argali were characterized by steep slope angle,shallow snow cover,high plant density,low bush number,low Stipa sp. density, few Ceratoides arborescens near hiding spots,and high hiding cover. Results of Principal Component Analysis(PCA) showed that the first 6 principal components explained 80.09% and 82.58% of the total variance among all bedding site variables on Mt. Kekesen and Mt. Kalamaili. PCA indicated that snow depth,slope angle,number of plant families, number of plant species,and number of bush species were important factors affecting bed-site selection by argali. PCA also
    indicated that hiding cover,altitude ,distance to water resources and distance to human settlements were important factors affecting bed-site selection by argali.
    Feeding and bedding habitat selection by red deer (Cervus elaphus alxaicus) during winter in the Helan Mountains,China
    LIU Zhensheng,ZHANG Mingming,LI Zhigang,HU Tianhua,ZHAI Hao
    2009, 29(2):  133-141. 
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    Winter feeding and bedding habitat selection by red deer (Cervus elaphus alxaicus) were studied from December
    2007 to January 2008 in the Helan Mountains of Ningxia and Inner Mongolia. We identified 72 feeding sites and 59 bedding sites by observation of fresh sign and direct observation of animals;for comparison,we identified 131 sites from systematically placed transects on which we observed no recent evidence of deer use. At each plot, we measured 18 topographic and biological variables. Indices of selectivity for feeding were positive for sites with smooth undulating slopes,overstories characterized by Ziziphus jujube,Salix spp. , mixed species,and unforested Additional variables yielding positive selectivity indices at feeding sites included tree heights > 2. 5 m, shrub density >9 trees/100 m2 ,vegetation coverage > 20% ,tree density < 2 /100 m2, slopes <15°, aspects with both sun and shade,distance from nearest tree > 6 m,distance to nearest bare rock > 20 m,and hiding cover < 86%. Selectivity indices for bedding sites were positive for smooth undulating slopes,elevations of 1 600 -2 000 m, woodlands of < 2 /100 m2 characterized by Ziziphus jujuba,Salix spp., Juniperus rigida,mixed species,and unforested,slopes < 15° on southerly aspects,shrub density < 2/ 100 m2 ,shrub height < 0.2 m,distance from nearest shrub > 10 m,distance from nearest bare rock >20 m,vegetation coverage >20% ,and hiding cover < 86%. Feeding habitat selected by red deer had lower vegetation coverage and less hiding cover than bedding habitat. We fitted a logistic regression model describing feeding habitat selection:1.155 - 0.149 (tree density) -0.066 (vegetation coverage) + 0.190 (slope). The model correctly predicted 86. 8% of feeding sites. The model of bedding habitat was best fit by - 30.936 + 0. 494 (tree density) +0.257 (slope) - 0. 002 (elevation)+0.387(hiding cover),correctly predicting 95.8% of bedding sites. Food resources,hiding cover,and special landform characteristics of the Helan Mountains are important factors influencing feeding and bedding habitat selection by red deer.
    Behaviour tactics in overwintering Eurasian red squirrel in Liangshui Nature Reserve,China
    RONG Ke,ZONG Cheng,MA Jianzhang
    2009, 29(2):  142-151. 
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    Low temperature is the main limiting factor affecting the overwintering of animals. Present behavior research of
    red squirrels (Sciurus vulgaris) in winter lacks direct observations.  We made a focal observation on red squirrels in broadleaved Korean pine forests in Liangshui nature reserve of China, since October 31st,2007 to March 25th,2008. Technology of radio telemetry and snow trace was used to detect the overwintering behaviors of red squirrels responded to the low temperature. &nbsp;Eight red squirrels were captured then marked by radio collar and colored plastic collar. The activity rhythm of red squirrels was a unimodal pattern in winter.&nbsp; The delay of outgo time (Kendall &tau; test; n = 21,r = - 0.92,&tau; = - 0. 86,P &lt;0.0001) and decrease of activity while (Kendall &tau; test;n = 21,r = 0.80,&tau; = 0.68,P &lt; 0.0001)was notable accompanied with the temperature decline. The home range of red squirrels was clearly elliptical (2. 46 &plusmn; 0.09 ha) without overlaps. Red squirrels hoarded the pine seeds all over the home rage as the only food in winter. The retrieving behavior occurred in three retrieved-cache areas in the home range. The home range can be divided into three parts according to the position and use rate of the nests and retrieved-cache areas,i. e. the major area,in mid-segment of the home range;the minor area,near to the original broad-leaved Korean pine forests and the third area,far from the original broad-leaved Korean pine forests in the home range. The use ratesamong retrieved-caches areas were significantly different during winter (early winter,Kruskal-Wallis test;df = 2,x2 = 5.65,P = 0. 0594,mid winter,Kruskal-Wallis test;df = 2,x2 = 14.24,P =0.0008; late winter,One-Way ANOVA Test,df = 2,18,F = 7.00,P = 0.0056). The retrieving-caches areas in the major area (use rate in early winter,37.8 &plusmn; 7. 7% ; use rate in mid winter 80.5 &plusmn; 9.2% ; use rate in late winter 40.5 &plusmn; 2.7% ) and minor area (use rate in early winter,41.8 &plusmn;5.9% ;use rate in mid winter 14.8 &plusmn;9.2% ;use rate in late winter 37.8 &plusmn;4.9% ) were the main retrieving-caches areas. The rate of empty retrieve significantly increased (Kruskal-Wallis test;df =2;x2 = 16.60; P = 0.0002) as time went deep into winter. We supposed that the retrieving behavior reminded red squirrels of the cached-seeds by spatial memory. &nbsp;Red squirrels used nests (6 &plusmn; 1)dispersed near to the retrieved-cashes areas by turns to avoid capture of predators. &nbsp;More nests (6 &plusmn; 1) near the caches-areas were alternatively used by squirrels. The use of nests remarkably related with the use of retrieving-caches areas (Kendall &tau; test;the major area n = 217,r = 0.79,&tau; = 0. 6256,P = 0. 0003;minor area n = 171,r = 0.67,&tau; = 0.6186, P = 0.0006; third area n = 88,r = 0.74,&tau; = 0.6565,P =0. 0006). Red squirrels usually stayed over in the nearest nest after retrieving Our research suggested that multi-dense area retrieving and the relevant tactics of nests alternation were integrated response to low temperature and predators in winter.
    Caching selection by plateau pika and its biological significance
    LIU Wei,ZHANG Yu,WANG Xi,ZHAO Jianzhong,XU Qingmin,ZHOU Li
    2009, 29(2):  152-159. 
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    The study analyzed the placing of haypiles,the quantities of haypiles,and the plant species composition of haypiles
    in different habitats,and compared the relation between haypiles and habitats and food items in winter by observational and sampling methods. The results showed that pikas adopted a special way in which haypiles were placed on Lamiophlomis rotata or on Oxytropis kansuensis to protect them from rotting. The haypile quantities were different in three kinds of habitats. The mean was 29.00 ± 17.09 unit/ ha in the forbs meadow,16.33 ±9.50 unit /ha in the Elymus nutans meadow, and 12.67 ± 6.66 unit / ha in the Kobresia humilis meadow. The 5 largest haypiles included 37 plant species in the Kobresia humilis meadow. Preference indices were above 3 for 10 plant species,and the highest proportion for plant species were Oxytropis kansuensis (23.6% ) and Ajania khartensis (19.1% ). The similarity analysis results showed that,except for the second haypiles,the similarity coefficients between the different haypiles were high. The correlation analysis for plant species composition was positive and significant (r = 0.7323,df =6,P <0.05). The enclosed haypiles were not consumed by pikas. Based on these results,we explained the biological significance of caching haypiles by pikas,and we think that pikas collected haypiles as a hedge against an unpredictable food shortage in winter.
    Dispersal and hoarding on acorns of Quercus variabilis by rodents in dif- ferent habitats
    ZHAO Xuefeng,LU Jiqi,QIAO Wangtie,TANG Fayou
    2009, 29(2):  160-166. 
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    We studied the fate of tagged acorns of cork oak (Quercus variabilis),which were released in three different
    types of habitats,secondary forest,shrub,and rehabilitated land,in Wangwu Mt. area in Jiyuan,Henan,China. In mid-
    September 2007,15,10,and 8 plots were set in secondary forest,shrub,and rehabilitated land,respectively. Forty tintagged acorns were placed in each plot. Checking and recording on the released acorns was,at two-day intervals,carried out till the end of field investigation,namely,62 days after the acorns were released. SPSS for Windows (version 13.0) was used for data analysis and statistics. The results of this study showed that: 1)Apodemus peninsulae,Niviventor confucianus, A. agrarius,and Sciurotamias davidianus are the principle rodent species that predate acorns of cork oak; 2)The acorn-removal rate in forest habitat was lower than that in shrub and rehabilitated land,while seed the consumption rate by rodents in shrub and rehabilitated land was higher than that in secondary forest; 3)The median survival time was significantly different among three habitats;4)The average dispersal distances were not significantly different among these habitats, while the differences between eaten and hoarded acorns were significant in each type of habitat. Our results indicated that habitat is one of the key biotic factors influencing seed survival and subsequent seedling establishment.
    Seasonal variations in densities of 5-hydroxytryptamine immunoreactive cells in the gut of Mongolian gerbils (Meriones unguiculatus)
    ZHANG Zhiqiang,LIU Quansheng,WANG Dehua
    2009, 29(2):  167-171. 
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    To explore the relationship between the density of 5-Hydroxytryptamine (5-HT) cells and seasonal environmental
    conditions,localizations and comparisons of 5-HT-immunoreactive (IR)cells in the five portions (duodenum,small intestine, caecum,colon and rectum) of the gut in Mongolian gerbils (Meriones unguiculatus) were studied in both late November (winter) in 2003 and July (summer)in 2004, respectively,using an avidin-biotin-peroxidase complex immunohistochemical method. 5-HT-IR cells were distributed mainly in the basal portion of the epithelium of the gut and epithelium of the gut gland,and showed similar morphological features both in winter and in summer. Most of 5-HT-IR cells were round or oval in shape,while fewer were long-bar,cone,or irregular in shape. In winter,the number of 5-HT-IR cells was the highest in the small intestine,fewer in the duodenum,caecum and rectum,and the lowest in the colon. In summer,the highest densities of 5-HT-IR cells were located in the duodenum and small intestine,densities were lower and similar in the other portions of the gut. The distribution of 5-HT-IR cells in different seasons may be related to the seasonal changes of food components. Except for the duodenum and colon,the cell densities of the other portions of the gut in winter were significantly higher than those of summer,which is beneficial for enhancing digestion and assimilation,may be an adaptive strategy to deal with differences in food quality and the increased food intakes needed to survive winter successfully. Both morphological features and seasonal dynamics of 5-HT-IR cells suggested that,at the cellular level,the gut of Mongolian gerbils can respond to complex environmental conditions,and this has adaptive regulation functions
    .
    Age-dependent variation in densities of somatostatin and substance P-immuno- reactive cells along the gut in the Mongolian gerbils (Meriones unguiculatus)
    ZHANG Zhiqiang,LIU Quansheng,WANG Dehua
    2009, 29(2):  172-177. 
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    Gastrointestinal hormones play important roles in regulating the digestive processes of small mammals,and
    change with age. Substance P (SP)and somatostatin (SS)immunoreactive (IR)cells have opposite functions:the former promotes motility of the small intestine,and the latter acts as an inhibitor. To clarify age-dependent variation in distribution, as well as interactive features of SS and SP-IR cells along the gut in the Mongolian gerbils (Meriones unguiculatus), localization and comparison of both endocrine cells in the five portions (duodenum,small intestine,caecum,colon and rectum)of the gut in young,adult,and old gerbils were studied in winter using avidin-biotin-peroxidase complex immunohistochemical method. Both endocrine cells showed similar morphological features at different ages,being round,oval, shuttle,cone,or irregular in shape. The distribution of SS-IR cells tended to be narrower with increased age. SS-IR cells were found mainly in the duodenum and small intestine for all ages,and no immunoreactivity was found in the colon and the rectum in old gerbils. In the caecum,old gerbils showed higher densities of SS-IR cells than did young and adult gerbils, but no significant differences were found in any other portion of the gut. SP-IR cells displayed the highest densities in the colon,caecum,and rectum for young,adult and old gerbils,respectively,and no immunoreactivity was found in the colon and the rectum for adult gerbils. Different from SS-IR cells,SP-IR cells displayed significant differences for the each portion of gut for different ages. In conclusion,SS and SP-IR cells showed different distributions and age-dependent developmental features along the gut of Mongolian gerbils,which may be related to food quality and the opposing physiological functions of the two endocrine cells.
    Morphological variation of the plateau zokor (Myospalax baileyi) from different geographical populations
    TANG Lizhou,ZHANG Tongzuo,SU Jianping
    2009, 29(2):  178-184. 
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    The variation of seventeen morphological traits for plateau zokor (Myospalax baileyi) collected from twelve geographical populations was analyzed based on principal component analysis and cluster analysis. The results of principal component analysis showed that the cumulative contributive proportions of the first three principal components for males and first four components for females were 78.483% and 79.587% respectively. The skull characteristics included profile length, basal length,and occipital width and otherwise,coupled with the least distance between the parietal ridges and the least distance between the frontal ridges representing the extent of zygomatic expanding might become the most contributive factors for the morphological variation among populations. The results of principal component analysis and cluster analysis congruously revealed that the populations were divided into two large regional groups:the Gansu group and the mixed group including
    Qinghai and Sichuan populations. These results actually reflected two different directions of evolution:one showed a lesser skull,divided parietal ridges and frontal ridges,such as Gansu group;another showed a larger skull,closed parietal ridges and frontal ridges,such as the mixed group.  This morphological variation of regional groups may result from the selection of environmental factors in the subterranean burrow system.
    Application of DNA pooling in combination with DHPLC and direct se- quencing in SNPs detection of the finless porpoise (Neophocaena phocaenoides)
    LI Shuzhen,WAN Huirong,YANG Guang
    2009, 29(2):  185-190. 
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    Denaturing high performance liquid chromatography (DHPLC) and direct sequencing in combination with DNA
    pooling were used as possible strategies for discussing the least rare allele frequency of DNA pools for SNPs detection in the finless porpoise (Neophocaena phocaenoides). To evaluate the utility and sensitivity of this approach,we constructed DNA pools comprised of 20 previously genotyped individuals with a frequency representation of 0% - 50% for the variant allele. The SNPs can be detected out at a frequency that no less than 5% when using DNA pooling and DHPLC. In contrast, fluorescent sequencing detected variants in the same pools only if the frequency of the less common allele was at least 10% . We conclude that the sample number of DNA pooling for DHPLC analysis should be no more than ten to ensure the efficiency and accuracy of SNPs discovery and screening. The high efficiency and accuracy of the present methods will make them effective in large-scale SNPs screening.
    Cloning and expression of HSF1 cDNA from Hainan Eld’s deer
    CHENG Ying,DU Li,WANG Fengyang,LIU Tao,LI Zhishen,XU Shiying,FU Yunnan,LIN Xianmei,WU Kebang,LIN Jiecai,MANCHU Riga
    2009, 29(2):  191-197. 
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    Heat shock transcription factor 1 (HSF1)cDNA of Hainan Eld’s deer (Cervus eldi hainanus) was amplified
    by RT-PCR and RACE,and cloned into pMD20-T vector. The recombinant plasmid was confirmed by PCR,endonuclease digestion,sequencing and bioimformatics analysis. The PCR product was then subcloned into pET28a vector and transformed into E. coli host cells. Protein expression was induced by IPTG and analyzed by SDS-PAGE and Western blot. The results showed that the HSF1 cDNA was 2 036 bp in length and contained a 1 578 bp ORF encoding 525 amino acids. The encoded protein was a hydrophilic protein which IP was 4. 93 analyzed by bioimformatics. A 62 kD fusion protein with a His tag was induced by IPTG and confirmed by Western blot using anti his tag monoclonal antibody,getting a special antibody binding band,indicated that prokaryotic expression vector of HSF1 from Hainan Eld’s deer was constructed and expressed successfully.
    Prokaryotic expression and polyclonal antibody preparation of plateau pika HIF-1α protein
    LI Hongge,ZHAO Xinquan,CHANG Zhijie,REN Fangli
    2009, 29(2):  198-203. 
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    The plateau pika (Ochotona curzoniae) is a high hypoxia-tolerant and cold adapted species living only at 3 000
    - 5 000 m above sea level on the Qinghai-Tibetan Plateau. Hypoxia inducible factor-1 (HIF-1),which is composed of
    HIF-1α and HIF-1β subunits,is a key transcription factor that regulates a variety of cellular and systemic adaptations to hypoxia. HIF-1 transcription activity is mainly regulated through oxygen-dependent modification of HIF-1α protein. The gene sequence encoding pika HIF-1α (pHIF-1α) (538/822) was inserted into PGEX-4T-1 bacterial expression vector using recombinant DNA techniques. After transformation by the expression vector, E. coli BL21 was induced to highly express the target protein fused by GST protein.  After identification and purification,the solubly expressed GST-pHIF-1α (538/822) fusion protein was used to immunize rabbits for preparing polyclonal antibody. The quality of the antibody was identified by Western blot. The results indicated that the polyclonal antibody recognizes plateau pika HIF-1α protein. The polyclonal antibody was further affinity purified. Western blot and immunostaining analysis reveal that the purified antibody can detect in vitro and in vivo expression and localization of plateau pika HIF-1α protein and be used in the further study.
    Co-expression of T’TB antigen epitope of canine distemper virus and VP2 protein of canine parvovirus and determination of specific antibodies in immunized mice
    WANG Yajun,HUA Yuping
    2009, 29(2):  204-209. 
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    The VP2 gene of canine parvovirus (CPV) was amplified by applying the PCR approach. The amplicon was ligated
    into the transfer vector pFastBacHTc of Bac-to-Bac baculovirus expression system. The vector carrying VP2 gene was named pFastBacHTc-VP2. The Artificially synthesized T’TB antigen epitope gene of canine distemper virus (CDV) was inserted into the vector pFastBacHTc-VP2 at the upstream position of VP2 to create a vector carrying both VP2 gene and T’ TB gene named pFastBacHTc-T’TB-VP2. pFastBacHTc-T’TB-VP2 was transferred into E. coli DH10Bac that contains baculovirus shuttle vector bacmid to obtain recombinant transfecting plasmid Bacmid-BacHT-T’TB-VP2. Bacmid-BacHTT’TB-VP2 was then used to transfect Sf-9 insect cells to express recombinant protein T’TB-VP2. Western blot analysis indicated that the molecular weight of T’TB-VP2 was 70 ku and could react with CPV and CDV antiserum. Purified T’TBVP2 protein was used to challenge BALB /c mice at the age of 6 -8 weeks without adjuvant. ELISA testing indicated that T’TB-VP2 protein was able to induce high level of antibodies specifically neutralizing CDV and CPV. This study laid a foundation for developing the new sub-unit vaccine of canine distemper and canine parvovirus.
    Diurnal behaviors and time budget of a female Yangtze finless porpoise during pregnancy
    SHU Jiakuan,JIANG Wenhua,ZHENG Bangyou,HAN Demin,SHANG Qiliang,YU Daoping
    2009, 29(2):  210-215. 
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    From August 25,2006 to August 15,2007 in the open water of Tongling Freshwater Porpoise National Nature
    Reserve,the diurnal behavior and time budget of a pregnant Yangtze finless female porpoise (Neophocaena phocaenoides asiaeorientalis) was observed using focal-animal and continuous sampling methods. The results indicated that: 1) the female predominantly invested its time in feeding (66 0 ±1 1% ,Mean ± SE),followed by traveling (11.5 ± 0.3% ),resting (10.5 ±0.5% ),socializing (9.8 ± 1.1% ) and other (2.1 ± 0.1% ), 2) among different months,it showed significant differences for each of the above behaviors except traveling, 3) each of the five behaviors was significantly different among the 11 periods of the day,and 4)significant differences were observed for each of the five behaviors across the five zones of the water.
    Habitat selection of masked palm civet in Houhe Nature Reserve,Hubei
    WANG Jian, LIU Qunxiu, TANG Dengkui, JIANG Guanghua, SUO Jianzhong, ZHENG Zhizhang,GONG Renhu,WANG Yongchao,HUA Yuping
    2009, 29(2):  216-222. 
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    We documented the activity tracks of masked palm civet (Paguma larvata) using radio-tracking and line-transects
    sampling methods from April to October of 2005. Sample plots (10 m × 10 m)were centered on tracks found according to radio-tracking signals. Line transects were set randomly throughout the research area,and sample plots (10 m ×10 m)were set at 150 m interval along line transects. Habitat selection by masked palm civet was studied by comparing the habitat properties recorded in both active (used by masked palm civet)and in active (unused by masked palm civet)sample plots. Principal components were analyzed for the habitat utilization of masked palm civet. In spring,canopy cover, distance to river,slope position,herbage cover and habitat type serve as important factors influencing habitat use by masked palm civet,and high canopy cover, proper distance to river,middle and higher slope position. In summer,distance to river served as the most important factor influencing habitat selection by masked palm civet,and slope position, vegetation types and canopy also were selectively used. In autumn,canopy cover,shrub cover and slope position were important for masked palm civet. Sunny slopes and high canopy cover were consistently used by masked palm civet,and slope position used was significantly different in different seasons. There were certain seasonal changes in slope position,distance to residents,herbage cover and distance to river used by masked palm civet.
    Comparison of hair trace elements content in different age groups of semifree-ranging cheetah (Acinonyx jubarus)
    WU Feng,SUN Qiang,ZHOU Yongxiao,XU Jianzhong,XU Chunzhong,XIE Chunyu,WU Kun,JIN Xiaojun
    2009, 29(2):  223-226. 
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    Five trace elements,including Cu,Zn,Pb,Mn,Se,were determined in the hair of 50 individual cheetahs (Acinonyx jubarus). Cu, Zn were determined by flame atomic absorption spectrophotometry (FAAS); Mn, Pb were determined by graphite furnace atomic absorption spectrometry (GFAAS)while Se was determined by atomic absorption spectrometry (AAS). The corresponding amounts of trace elements are significantly different according to different genders. The Cu and Zn amounts in the hair of different age groups differ significantly. The zinc amount increases continuously from sub-adult group to adult group with a peak in the adult group and then a slight decrease in the senior group. The copper amount increases with age. Manganese and lead amounts of different age groups differ little,but tend to increase with age. The selenium amounts of different age groups differ significantly with the peak in the adult group.
    Mammalian skeleton specimen preparation using proteolytic enzyme (alcalase)detergent
    WU Yonghua,YANG Qisen
    2009, 29(2):  227-230. 
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    We report on an easy method to remove muscle tissue from the skeleton by alcalase detergent in mammalian skeleton specimen preparation. In the present paper,using hare skull specimen as an example,we describe the alcalase detergentmethod in details and compare it with other methods for mammalian skeleton specimen preparation. In this method,hare muscle tissues are removed from skeletons by alcalase detergent. With commercial proteinases,alcalase (2.4 L, 2.4 AU/ g, Novozymes Biotechnology Co. , Ltd. ), the process requires around 10 minutes’skeletal simmering in a 0.5% solution (V/V) of the alcalase and water under 60 ℃ condition. In total, about 101 rat skull and 487 hare skull specimens are made using this method. Compared to other traditional methods in mammalian skeleton specimen preparation,the alcalase detergent method is safer,more efficient and less costly,and it is also suitable for batch production of skeleton specimen.