The complete mitochondrial genome of the domestic red deer (Cervus elaphus) of New Zealand and its phylogenic position within the family Cervidae

We determined the complete nucleotide sequence of the mitochondrial genome of the semidomestic red deer (Cervus elaphus) of New Zealand. The genome was 16 357 bp long and contained 13 protein‐coding genes, 12SrRNA, 16SrRNA, 22 tRNAs and a D‐loop as found in other mammals. Database homology searches showed that the mitochondrial DNA (mtDNA) sequence from the New Zealand semidomestic deer was similar to partial mtDNA sequences from the European, Norwegian (C. e. atlanticus) and Spanish red deer (C. e. hispanicus). Phylogenetic analysis of the mitochondrial protein‐coding regions revealed two well‐defined monophyletic clades in subfamilies Cervinae and Muntiacinae. However, red deer and Sika deer were not found to be close relatives. The analysis did identify the red deer as a sister taxon of a Samber/Sika deer clade, although it was more closely related to the Samber than the Sika group.     

Detection of nucleolar organizer regions on chromosomes by flourescence in situ hybridization with human 28S rRNA gene and cloning of 28S rRNA gene in Sika deer

The number and loci of nucleolar organizer regions (NOR) on chromosomes in Sika deer (Cervus nippon centralis) were determined by fluorescence in situ hybridization with a human 28S ribosomal RNA (rRNA) gene as a probe. Sika deer that live in Nikko National Park and its neighboring areas (Asio and Seta) in Japan were used. All of the analyzed metaphases had three or four NOR at the end of the first and second longest telocentric autosomes. Nucleolar organizer region association, which is associated specifically on parts of NOR between chromosomes, was also observed clearly. A Sika deer 28S rRNA gene was produced by a polymerase chain reaction method. The nucleotide sequence of a Sika deer 28S rRNA gene determined by an automatic sequencer was 97 bp, and showed homogeneity of 88% for the human sequence.     

Sex and age differences in meat composition of Yeso sika deer (Cervus nippon yesoensis) reared for a short period after capture in the wild

Yeso sika deer captured in winter around Lake Akan in Hokkaido were reared for 8–10 months at Tokyo University of Agriculture in Abashiri. Six 1‐year‐old females and males and six 2‐year‐old or older (adult) females and males were slaughtered and their carcasses were processed. The chemical composition, mineral contents and fatty acid composition of the loin were measured. No marked influence of gender or age was noted in the chemical composition of loin. In the mineral contents, significant differences were noted. The potassium and sulfur contents were lower and the sodium content was higher in adult deer meat (P < 0.05, respectively) and the potassium content was higher in male deer meat (P < 0.05). Arsenic, cadmium or lead were not detected. In the unsaturated fatty acid, a significant interaction was detected (P < 0.05), and it was high in 1‐year‐old female deer meat and low in 1‐year‐old male deer meat. Significant gender or age differences were noted only in the mineral contents in the loin of deer reared for a short period after capture.     

梅花鹿茸鲜重与年龄的相关性研究及经济效益分析

以江苏地区的梅花鹿为对象,研究分析了梅花鹿鹿茸重量与其年龄之间的相关性。结果表明:9龄内梅花鹿的鹿茸重与年龄呈正相关(r=0.767,P<0.05),它们之间线性回归方程为y=0.345x+1.186。梅花鹿3～8锯的鲜茸单产很高,平均水平达3.28kg/锯。公鹿鲜茸重的变异系数在2～6龄之间逐渐减小,7龄之后又有上升的超势,其中以5～7龄的变异系数最小,分别为16.2%、6.1%、11.7%。相关数据分析表明,梅花鹿公鹿在1～4龄内经济效益较差,从5龄开始获利,年获利逐年增加,这种趋势与梅花鹿的茸产量变化趋势是一致的。     

Seasonal changes of the mineral contents in the rumen of wild Yeso sika deer (Cervus nippon yesoensis)

The rumen contents were collected from 36 wild Yeso sika deer (Cervus nippon yesoensis) captured by deer culling or by hunting in the spring, summer, autumn and winter in Hokkaido, Japan. Botanical classification was conducted, and the contents of mineral (calcium (Ca), phosphorus (P), potassium (K), sodium (Na), iron (Fe), copper (Cu) and zinc (Zn)) were measured. The animals were captured around pastures or fallow field areas in the Kushiro area. The rumen contents consisted of grasses and Sasa sp. leaves regardless of the season. Leaves and bark were ingested in the spring, autumn and winter. The macro‐mineral contents in the rumen showed seasonal changes. In the summer, the Ca, K and P contents were high, and the Na content was low. There were no seasonal changes in the Fe content. The P, Na and Fe contents were higher than the animals' requirements. In a future survey, it is needed to determine the mineral contents of the food ingested by wild Yeso sika deer.     

Comparative digestion of cell wall components of alfalfa hay cubes between Sika deer (Cervus nippon) and cattle

Four female Sika deer (mean bodyweight, 48 kg) and three male Holstein cattle (mean bodyweight, 209 kg) were offered alfalfa hay cubes at 2% (deer) and 2.5% (cattle) of bodyweight, respectively. The digestibility of the cell walls and cell wall components (rhamnose, arabinose, xylose, mannose, galactose and glucose) in alfalfa and its retention time of the alfalfa in the digestive tract and rumen parameters were determined. Cell walls and xylose were less digestible in the deer than in the cattle (P < 0.01 and P < 0.05, respectively). The digestibility of galactose in the deer was as high as that in the cattle. The digestibility of the other sugars and total neutral sugars was numerically lower in the deer, but the differences were not significant. In the deer, mannose was most digestible, followed in order by galactose, arabinose, glucose, rhamnose and xylose; whereas, in the cattle, mannose was most digestible, followed in order by arabinose, glucose, galactose, rhamnose and xylose. The retention time in the digestive tract was shorter in the deer than in the cattle. In the deer, the number of ruminal protozoa was somewhat higher, and the concentrations of propionic acid and butyric acid were higher (P < 0.05) than in the cattle. These results indicate that Sika deer might utilize pectic polysaccharides as a carbon source in preference to glucose containing polysaccharides such as cellulose. The lower digestibility of all cell wall components except galactose in the Sika deer might be mainly due to its shorter retention time.     

Effects of the feeding of wild Yeso sika deer (Cervus nippon yesoensis) on the prevention of damage due to bark stripping and the use of feeding sites

Feeding sites for wild Yeso sika deer around Lake Akan, Japan, were established. Effects on the number of deer using the feeding sites, the prevention of bark stripping damage, the amount of feeding, and eating time in a 5‐year period (1999–2003) were evaluated. The number of deer using feeding sites increased with years during the feeding period. The damaged tree ratio after the initiation of feeding markedly decreased compared with 16.5% before the initiation of feeding. After the start of feeding, there were no trees with damage the entire circumference. According to tree species, the number of damaged trees of Ulmus laciniata Mayr as a percentage of all investigated trees was high (5.2%). The total amount of beet pulp feeding increased with the feeding year, showing 4.5‐fold increase. At feeding sites in deer culling, eating behavior was observed during the night. The preventive effects on bark stripping damage continued during the 5‐year feeding period. However, with the course of feeding years, the number of deer using feeding sites and the level of feeding increased.     

Determination of nucleotide sequence of SRY gene in sika deer (Cervus nippon)

The aim of the present study was to determine the whole nucleotide sequence of the open reading frame of the sex‐determining region Y (SRY‐ORF) in wild sika deer. The SRY gene of wild sika deer was obtained by polymerase chain reaction (PCR) with DNA from blood samples. The whole nucleotide sequence of the SRY‐ORF in wild sika deer consisted of 687 bp and encoded 229 deduced amino acids. In comparison with the bovine SRY gene, the percentage of nucleotide sequence homology was 91.0% in the overall ORF, and those of the N‐terminal, high mobility group (HMG) box, and C‐terminal regions within ORF were 88.9%, 96.2% and 87.9%, respectively. The nucleotide sequences of sika deer SRY‐ORF characterized in the present study can be used for phylogenetic analysis or sexing in wild sika deer.     

长春地区梅花鹿毕氏肠微孢子虫分子流行病学调查

为了掌握长春地区梅花鹿毕氏肠微孢子虫(Enterocytozoon bieneusi)的感染率及基因型种类的分布情况,探究其在人与动物之间传播的源由,本研究于2018年12月-2019年12月期间采集长春地区8个养殖场共计538份梅花鹿粪便样品进行毕氏肠微孢子虫的流行病学调查及基因型分析,经PCR鉴定,测序分析、序列拼接比对以及系统进化分析。试验结果表明,长春地区梅花鹿毕氏肠微孢子虫的感染率为17.84%(96/538),不同养殖场的感染率为0~47.96%;研究共发现14种基因型(BEB6、EbpC、Ⅰ、JLD-Ⅲ、JLD-Ⅸ、JLD-ⅩⅤ、JLD-ⅩⅥ、JLD-ⅩⅦ、JLD-ⅩⅧ、JLD-ⅩⅨ、JLD-ⅩⅩ、JLD-ⅩⅪ、JLD-ⅩⅫ和JLD-ⅩⅩⅢ),其中BEB6为优势基因型。应用多位点序列分型(MLST)方法,选择MS1、MS3、MS4以及MS7位点对毕氏肠微孢子虫阳性样本进行多态性和群体遗传结构分析,结果显示,在96份阳性样品中,共有16份阳性分离株在至少2个位点同时扩增成功,形成10个多位点基因型(multilocus genotypes,MLGs),相同的基因型之间存在不同亚型,表明长春地区梅花鹿毕氏肠微孢子虫具有丰富的遗传多样性。本研究应用分子生物学方法对长春地区梅花鹿毕氏肠微孢子虫的流行病情况及基因型进行分析,研究结果不仅为梅花鹿毕氏肠微孢子虫防控提供科学依据,也为防控毕氏微孢子虫在人和动物间的传播提供基础数据,对保障公共卫生与健康意义重大。     

梅花鹿S100A16基因的克隆及生物信息学分析

为探索梅花鹿（Cervus nippon）S100A16基因序列及生物学特性，本研究根据GenBank数据库中牛、绵羊S100A16基因序列设计引物，以梅花鹿鹿茸顶端组织cDNA为模板，采用RT-PCR技术和分子克隆技术成功获得梅花鹿S100A16基因的cDNA序列。生物信息学分析发现，梅花鹿S100A16基因CDS区全长312 bp，编码103个氨基酸；蛋白含有11个磷酸化位点，有跨膜结构域，无信号肽，为在细胞内发挥作用的稳定蛋白；蛋白仅在C端含有S100蛋白家族经典的EF螺旋结构域，由12个氨基酸组成，N端EF螺旋结构域由15个氨基酸组成；蛋白C端含有FGF-1蛋白结合位点；梅花鹿S100A16蛋白的二级结构主要由α-螺旋和无规则卷曲构成；三级结构显示该蛋白有2个Ca²⁺结合位点；梅花鹿S100A16蛋白氨基酸序列与东欧马鹿同源性最高，为100%，与其他部分物种S100A16蛋白氨基酸序列构建系统进化树，分析表明S100A16基因在进化上比较保守，符合功能基因的特点。研究结果为进一步揭示梅花鹿S100A16基因的功能及表达机制提供依据。     

鹧鸪线粒体COII、ATPase8和3个tRNA基因序列分析及物种鉴别研究

线粒体DNA是真核生物核外遗传物质,其结构简单,发生变异的机率相对稳定,被广泛用于系统起源、进化、亲缘关系及物种鉴别的研究中。为了挖掘禽类（源性成分）鉴别的分子生物学数据,探讨线粒体不同基因发育信息及鉴别的可信度和灵敏度。本研究扩增鹧鸪线粒体基因组中COⅡ、ATPase8区域,克隆出1138 bp片段,测序结果分析表明,在所克隆的片段中,含有COⅠ基因部分序列、tRNA^Ser基因、tRNA^Asp基因、COⅡ基因、tRNA ^Leu基因、ATPase8基因全序列和ATPase6基因部分序列。分析COⅡ、ATPase8和tRNA基因显示其与鹌鹑、鸡、火鸡、鹅、鸵鸟5种禽类的序列均具有较高的同源性,基于它们MEGA3.1构建系统进化树,探讨了它们所含的信息量及在鹧鸪和其它禽类鉴别方面的应用潜力,发现COⅡ比ATPase8和tRNA基因含有更多的鉴别信息。     

Mitochondrial DNA variation and genetic relationships in Spanish donkey breeds (Equus asinus)

In this study, the mitochondrial DNA diversity of six Spanish donkey breeds and two African donkey populations (one from Morocco and the other from Zimbabwe) was analysed. A total of 79 animals were sequenced for 313 bp of the cytochrome b gene, and 91 individuals for 383 bp of the D‐loop region or control‐region. Sequence comparisons and phylogenetic analyses of both Spanish and African populations revealed low diversity. Only six and seven haplotypes respectively were found in cytochrome b and the D‐loop region. Relatively low nucleotide diversity (π) values were detected in these populations. The π values, from the D‐loop region, ranged from 0.0006 to 0.0169 for the Catalana and Andaluza breeds, respectively. The obtained results seem to confirm the existence of two divergent maternal lineages of African origin (Equus asinus africanus and E. a. somaliensis). In this paper the genetic relationships between these breeds are analysed and compared with those obtained in other European populations. Also, the data on the genetic relationships between the populations, obtained from nuclear DNA (microsatellites) and mitochondrial DNA (cytochrome b and D‐loop region) is argued and interpreted.     

Functional responses of cougars (Puma concolor) in a multiple prey‐species system

The study of predator–prey interactions is commonly analyzed using functional responses to gain an understanding of predation patterns and the impact they have on prey populations. Despite this, little is known about predator–prey systems with multiple prey species in sites near the equator. Here we studied the functional response of cougars (Puma concolor) in relation to their main prey, armadillo (Dasypus novemcinctus), coati (Nasua narica) and white‐tailed deer (Odocoileus virginianus). Between 2004 and 2010, cougar scats were collected along 5 transects to estimate the consumption of different prey species. A relative abundance index (RAI) was calculated for each prey species and cougar using 18 camera traps. We compared Holling type I, II and III functional response models to determine patterns in prey consumption based on the relative abundance and biomass of each prey species consumed. The 3 main prey species comprised 55% (armadillo), 17% (coati) and 8% (white‐tailed deer) of the diet. Type I and II functional responses described consumption of the 2 most common prey species armadillos and coati similarly well, while a type I response best characterized consumption of white‐tailed deer. A negative correlation between the proportions of armadillo versus coati and white‐tailed deer biomass in cougar scats suggests switching to consume alternative prey, confirming high foraging plasticity of this carnivore. This work represents one of the few studies to compare functional responses across multiple prey species, combined with evidence for prey‐switching at low densities of preferred prey.     

梅花鹿三种保定药物对比实验

本文通过用静松灵、司可林、眠乃宁三种保定药物保定梅花鹿对比实验，得出：①静松灵一次性给药剂量较大，生产中操作不方便。②司可林一次给药剂量虽小，但有效剂量与致死剂量十分接近，剂量误差易导致鹿死亡。③静松灵和司可林均无有效拮抗药物，且后遗性影响较大。④眠乃宁注射剂量适中，剂量误差不会导致鹿死亡，无后遗性影响。拮抗药物苏醒灵拮抗效果优良，使眠乃宁的应用更安全。     

Spiculopteragia mathevossiani Ruchliadev, 1948 is the minor morph of Spiculopteragia spiculoptera (Gushanskaya, 1931): molecular evidence

Although Spiculopteragia spiculoptera is primarily a parasite of cervids, it can also but less often contaminate domestic livestock. Little is known about its epidemiology and its pathogenicity in domestic ruminants and other unusual cervid species. Its taxonomic status remains unclear since the hypothesis of morphological polymorphism among males has been proposed. However, accurate taxonomy is fundamental in the identification and survey of potentially pathogenic species of parasites. The second internal transcribed spacer of rDNA (ITS-2) and the mitochondrial (mt) DNA-derived ND4 gene were used to study the polymorphism hypothesis for S. spiculoptera. ND4 evolves more quickly than ITS-2 and is considered to be more discriminant in the characterization of closely related species. DNA sequences of ITS-2 and ND4 were studied in 18 individual males of morphological type spiculoptera and in 3 of morphological type of mathevossiani from Red deer (Cervus elaphus), Roe deer (Capreolus capreolus) and Chamois (Rupicapra rupicapra). Intraindividual ITS-2 variations were detected within and between each morphotype of Spiculopteragia but these differences did not separate the two morphs mathevossiani and spiculoptera. Similarly, although ND4 showed a high level of nucleotide substitution, the morphotypes S. mathevossiani and S. spiculoptera were clustered together. Our genetic data support the dimorphic male hypothesis for the species S. spiculoptera.     

某鹿场梅花鹿种群退化的调查

调查了 1988～ 2 0 0 1年间黑龙江省平山养鹿场梅花鹿种群生物学特性变化特点。结果表明 ,仔鹿病死率增加 12 4 6 6 % ,成年鹿病死率增加 12 3 12 % ;母鹿产仔率减少 10 0 4 % ,2 0d仔鹿成活率减少 8 5 7% ;公鹿鹿茸平均单产降低 2 8 5 7% ,一等品率降低 5 2 2 3% ,等外品率增加 12 5 14 %。分析该结果原因为近亲繁殖所致。     

Phylogenetic relationship and variation of alarm call traits of populations of red‐cheeked ground squirrels (Spermophilus erythrogenys sensu lato) suggest taxonomic delineation

Distribution area and taxonomic borders within the species complex Spermophilus erythrogenys sensu lato remain questionable. Early evidence suggests that red‐cheeked ground squirrels of Southeast Kazakhstan are remarkably different in terms of the acoustic structure of their alarm calls from the red‐cheeked ground squirrels of the Kurgan region in Russia. In this study, we analyzed the differences in the acoustic structure of the alarm call and mitochondrial DNA (complete control region, 1005–1006 bp and complete cytochrome b gene, 1140 bp) in 3 populations of red‐cheeked ground squirrels (Tara, Altyn‐Emel and Balkhash), all located within areas isolated by geographical barriers in Southeast Kazakhstan. We found that the alarm call variables were similar between the 3 study populations and differed by the maximum fundamental frequency (8.46 ± 0.75 kHz) from the values (5.62 ± 0.06 kHz) reported for the red‐cheeked ground squirrels from the Kurgan region of Russia. Variation in mtDNA control region was only 3% and variation in cytochrome b gene was only 2.5%. Phylogenetic trees based on cytochrome b gene polymorphism of 44 individuals from the study area and adjacent territories indicated 3 clades with high (98–100%) bootstrap support: “intermedius,” “brevicauda” and “iliensis”). We conclude that the 3 study populations in Southeast Kazakhstan belong to the clade intermedius and suggest a taxonomical revision of the species complex Spermophilus erythrogenys sensu lato, including analyses of nuclear DNA and alarm calls for populations of the brevicauda and iliensis clades.     

东北梅花鹿优化育种规划步骤、基本条件及对现行育种方案的分析

对东北梅花鹿优化育种规划的具体步骤和所应具备的基本条件进行了研究 ,并在分析东北梅花鹿群现状的基础上 ,得出群体的经济学和遗传参数、生物学和育种技术指数、群体结构参数、育种成本与投资参数等一系列将在育种规划计算中应用的参数 ,同时还对东北梅花鹿现行育种方案的育种效果进行了分析。     

牛病毒性腹泻病毒梅花鹿分离株E0基因的克隆与表达

利用RT-PCR技术扩增了牛病毒性腹泻病毒(BVDV)梅花鹿分离株E0基因，并将其克隆到pMD18-T克隆载体和pET28a原核表达载体中，构建了pMD18-T／E0和pET28a／E0重组子，并进行了测序和表达。表达产物分别用12％SDS-PAGE检测和牛抗BVDV阳性血清进行Western-blotting检测，结果表明．BVDV梅花鹿分离株E0基因的核苷酸序列与匈牙利VEDEVAC株的同源性高达98．6％，与C21V标准株的同源性为84．9％，证明构建的重组子是正确的；BVDV梅花鹿分离株E0基因得到了正确表达。     

Differentiating among horse (Equus caballus), donkey (Equus asinus) and their hybrids with combined analysis of nuclear and mitochondrial gene polymorphism

A novel and brief method of differentiating among horse (Equus caballus) and donkey (Equus asinus) and their hybrids (mule, E. asinus × E. caballus and hinny, E. caballus × E. asinus) with combined analysis of nuclear and mitochondrial gene polymorphism (CANMGP) was reported in the present report. A nuclear gene, protamine P1 gene of donkey was sequenced and compared with the known horse sequence from GenBank while a published equid mitochondrial gene, cytochrome b gene of donkey was compared with that of horse. In each of the two genes, a fixed nucleotide substitution within an exon that could be recognized by Dpn II restriction enzyme was found between the two species. Two pairs of primers were designed for amplifying the fragments within the two genes containing the informative nucleotide positions in 65 horses and 41 donkeys and 38 hybrids and conditions of polymerase chain reaction and restriction fragment length polymorphism (PCR‐RFLP) analysis were optimized. Horse, donkey and mule and hinny had their own specific cleavage patterns after the PCR‐RFLP analysis was performed, which made it very easy to identify them from each other. As multiplex PCR can be conducted with the two pairs of primers and only one restriction enzyme is involved in PCR‐RFLP analysis, the method described in the present study is a convenient way to identify horse and donkey and their hybrids. The idea involved in the method of CANMGP can be also used to differentiate other animal species or breeds and their hybrids.