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.     

Analysis of mitochondrial DNA protein-coding region in the Yeso Sika deer (Cervus nippon yesoensis)

In the present study, mitochondrial DNA sequences of the Yeso Sika deer (Cervus nippon yesoensis) were studied. Specifically, protein‐coding genes as mitochondrial NADH dehydrogenase subunits (ND1, ND2, ND3, ND4L, ND4, ND5 and ND6), cytochrome c oxidase subunits (CO I and CO III), ATP synthase subunits (ATPase8 and ATPase6) and cytochrome b. Also, phylogenetic analyses on eight mammalian species were performed, including the Muntjac deer (Muntiacus reevesi). The rate of amino‐acid substitution was lowest (3.74%) between Yeso Sika deer and Muntjac deer, and the values between Yeso Sika deer and other species (sheep, cattle, horse, pig, mouse, human and chimpanzee) were 6.63%, 7.30%, 12.55%, 13.03%, 23.59%, 24.82% and 25.04%, respectively. Among them, the highest value of divergence was recognized in ATPase8, and the second structure of ATPase8 showed a difference between the Yeso Sika deer and Muntjac deer as a result of the substitution of 34His→Tyr and 49Thr→Ile. In addition, we identified a substitution of an amino‐acid sequence (19Thr→Ala) between the Yeso Sika deer and Yakushima Sika deer (C. n. yakushimae). From these results, ATPase8 was also a variable region in Cervidae.     

A TaqMan real-time PCR-based assay for the identification of Fasciola spp

Real time quantitative PCR (qPCR) is one of the key technologies of the post-genome era, with clear advantages compared to normal end-point PCR. In this paper, we report the first qPCR-based assay for the identification of Fasciola spp. Based on sequences of the second internal transcribed spacers (ITS-2) of the ribosomal rRNA gene, we used a set of genus-specific primers for Fasciola ITS-2 amplification, and we designed species-specific internal TaqMan probes to identify F. hepatica and F. gigantica, as well as the hybrid 'intermediate'Fasciola. These primers and probes were used for the highly specific, sensitive, and simple identification of Fasciola species collected from different animal host from China, Spain, Niger and Egypt. The novel qPCR-based technique for the identification of Fasciola spp. may provide a useful tool for the epidemiological investigation of Fasciola infection, including their intermediate snail hosts.     

Functional anatomy of the omasum in high Arctic Svalbard reindeer (Rangifer tarandus platyrhynchus) and Norwegian reindeer (Rangifer tarandus tarandus)

The structure and fill of the omasum was investigated in summer and in winter in adult female reindeer living on the polar desert and tundra of the high Arctic archipelago of Svalbard and in sub-Arctic mountain habitats in northern Norway. The mean total mass of the omasum in non-lactating adult female Svalbard reindeer was 467 g (0.65 g per 100 g live body mass (BM)) in September and 477 g (1.03 g per 100 g BM) in April. By contrast, the mean mass of the omasum in non-lactating adult reindeer in northern Norway was 534 g (0.83 g per 100 g BM) in September but only 205 g (0.35 g per 100 g BM p < 0.05) in late March, owing to a decrease in both tissue mass and the wet mass of the contents of the organ. The mean absorptive surface of the omasum in Svalbard reindeer was 2300 cm2 in September and 2023 cm2 in April. In Norwegian reindeer, by contrast, the absorptive surface area decreased from 2201 cm2 in September to 1181 cm2 (p < 0.05) in late March. The marked seasonal decline of omasal tissue and contents in Norwegian reindeer probably results from intake of highly digestible forage plants, including lichens, in winter. Svalbard reindeer, a non-migratory sub-species, survive eating poor quality fibrous vascular plants in winter. The absence of any marked seasonal change in the mass, total absorptive surface area or filling of the omasum in Svalbard reindeer in winter despite a substantial decline in body mass presumably reflects their need to maintain maximum absorption of nutrients, including volatile fatty acids, when feeding on such poorly fermentable forage.     

Experimentally induced infection of reindeer (Rangifer tarandus) with Mycobacterium bovis.

In the USA, all species of Cervidae are included in the USDA's uniform methods and rules for the eradication of bovine tuberculosis and, therefore, are subject to regulations regarding intradermal tuberculin testing. In reindeer (Rangifer tarandus), infection with Mycobacterium bovis is exceedingly rare and the response of reindeer to infection with M. bovis in pathologic and immunologic terms is unknown. The objectives of the study reported here were to describe the pathologic changes associated with M. bovis infection in reindeer and evaluate the effectiveness of intradermal tuberculin testing as a means of diagnosis of tuberculosis in reindeer. Thirteen reindeer were inoculated intratonsilarly with 10(5) colony-forming units (CFU) of M. bovis, and 4 noninoculated reindeer served as negative controls. The comparative cervical test (CCT) was done on all reindeer 90 and 240 days after inoculation. Thirteen months after inoculation, all reindeer were euthanized and examined. All experimentally inoculated reindeer developed lesions in the medial retropharyngeal lymph nodes. The CCT accurately identified all M. bovis-inoculated reindeer, but false-positive results were common among negative-control reindeer. Modifications of the method for interpretation of the CCT decreased false-positive results without increasing false-negative results. Reindeer are susceptible to infection with M. bovis; however, lesions are fewer in number, less severe in nature, and less widely disseminated than are those seen in white-tailed deer (Odocoileus virginianus). Comparative cervical skin testing of reindeer can be highly sensitive, but has low specificity. Specificity can be improved by modification of criteria for interpretation of the CCT.     

Haematology of clinically normal and sick captive reindeer (Rangifer tarandus)

A retrospective analysis of haematological values from clinically normal captive reindeer (Rangifer tarandus) showed that the red cell count, haemoglobin level, packed cell volume and lymphocyte count were higher and the erythrocyte sedimentation rate and eosinophil count were lower in juveniles than in adults. Newborn animals were anaemic compared with juveniles and adults and had high reticulocyte counts. The values from healthy reindeer were used to identify abnormal haematological variations in a number of sick animals. It was shown that reindeer exhibit similar haemopathological responses to those of other artiodactyla, with increases in the erythrocyte sedimentation rate and fibrinogen level being of particular diagnostic significance. Eosinophilia was the only abnormal haematological finding in individuals with subclinical infections of intestinal parasites.     

A simple molecular method for discriminating common filarial nematodes of dogs (Canis familiaris)

Accurate diagnosis of canine filariosis is essential for choosing correct therapeutic approach. Therefore, reliable methods for discriminating among the different filarial infections in dogs are needed. The authors report simple and highly specific molecular methods that identify the three most common filarial nematodes of European dogs: Dirofilaria immitis, D. repens and Acanthocheilonema (syn. Dipetalonema) reconditum, based on (1) PCR amplifications of mitochondrial DNA (12S rDNA and coxI) with general filarial primers followed by digestion with restriction enzymes that generates band polymorphisms clearly discriminating the three species and (2) PCR amplifications with species-specific primers to support the restriction analysis, in particular in the case of multiple infections.     

基于线粒体Cyt b DNA阿拉善马鹿分子系统学研究

阿拉善马鹿为国家Ⅱ级重点保护野生动物,仅分布于宁夏和内蒙古交界的贺兰山中段,是我国唯一幸存的该亚种的有效种群。目前关于野生阿拉善马鹿的分子研究尚属空白,本研究于2016年8—9月和2017年2—3月在贺兰山采集新鲜阿拉善马鹿粪便样本396份,使用线粒体Cyt b区DNA引物对提取的DNA进行物种鉴定和9对微卫星引物对其个体识别,共鉴定出278个阿拉善马鹿个体,基于线粒体Cyt b DNA对其全序列进行分析。使用线粒体Cyt b区引物对提取的粪便DNA扩增,获得了1 075 bp的序列,检测到10个变异位点,并定义了10个单倍型。其中10个变异位点占分析长度的0. 93%,均为碱基代换,无碱基插入和缺失。其单倍型多样性为0. 243,核苷酸多样性为0. 000 32。中性检验Tajima's D值为显著负值,Fu和Li's D值为不显著的负值,表示阿拉善马鹿可能经历瓶颈事件并随后出现种群扩张。在系统发育树中,阿拉善马鹿单独聚成一支且与东北马鹿亲缘关系较近,与其他中国马鹿亚种亲缘关系较远。本研究表明野生阿拉善马鹿种群总体遗传多样性较低,建议加强对该亚种的保护力度。     

Detection of Ehrlichia muris DNA from sika deer (Cervus nippon yesoensis) in Hokkaido, Japan

Ehrlichia muris DNA was detected in the blood of sika deer (Cervus nippon yesoensis) by species-specific PCR based on the citrate synthase gene, which was shown to be more sensitive than species-specific PCR based on the 16S rRNA gene. Among 102 deer examined, one deer was positive. Deer may be a possible mammalian reservoir of E. muris.     

马鹿精子形态与超微结构研究

文中对马鹿(Cervus elaphus)精子形态与超微结构进行了研究。马鹿精子经固定、脱水、置换、包埋和聚合,用超薄切片机切片,再用醋酸双氧铀、柠檬酸铅染片,最后于透射电镜下观察其超微结构的变化。观察结果表明:马鹿精子全长(58.75±2.35)μm,由头部(8.93±0.24)μm、颈部(1.00±0.16)μm和尾部(48.18±1.18)μm三部分组成;头部呈扁卵圆状,绝大部分被浓缩且电子密度较高的精核所占据,顶体似帽状扣在精核之上,约占头部的2/3,其前部较为膨大;颈部位于头部和尾部之间,这个区域较短;尾部可分为中段、主段和终段。马鹿尾部轴丝的结构类型为"9+2";微管结构类型为"9+9+2"。     

Theileriosis in a reindeer (Rangifer tarandus tarandus) associated with a potentially novel Theileria sp

A 5‐year‐old male neutered reindeer (Rangifer tarandus tarandus) from Missouri was presented with a 3‐week history of anorexia, respiratory distress, lethargy, and weight loss. Blood smear review revealed that a small percentage of RBCs contained small (1–2 μm in length) pleomorphic piroplasms (signet ring, rod‐ or pear‐shaped, and elongate forms) with an eccentric magenta nucleus and basophilic cytoplasm. Nested PCR to specifically amplify a portion of the piroplasm small subunit ribosomal RNA (SSU rRNA) gene was performed on DNA extracted from an EDTA specimen of whole blood. Subsequent sequence analyses showed similarity between the reindeer hemoparasite and Theileria spp SSU rRNA gene sequences in the GenBank database, with highest similarity to those of a Theileria sp in a White‐tailed deer from North Texas (AY735132, AY735133). The reindeer and North Texas Theileria sp are genetically distinct from, albeit closely related to, the White‐tailed deer Theileria sp (subsequently referred to as T cervi). To the authors' knowledge, this is the first identification of Theileria of this genotype in a reindeer. Historically, T tarandirangiferis infection was found with associated mortality in reindeer in Russia, but reports predate molecular characterization. Hence, the relationship of T tarandirangiferis with either T cervi or this agent remains unknown. T cervi is not typically pathogenic in White‐tailed deer in the US unless the animal is immune‐compromised by stress or disease; however, mortality from T cervi infection in reindeer has been reported.     

Sex determination based on fecal DNA analysis of the amelogenin gene in sika deer (Cervus nippon)

A sex determination method using DNA extracted from feces has been developed for sika deer (Cervus nippon). We determined a partial sequence of the amelogenin gene of sika deer, which exists on both X and Y chromosomes with a deletion region on the Y chromosome. Based on the sexually dimorphic sequences, we designed a pair of primers which could amplify DNA fragments the lengths of which are different between males and females. PCR products were detected in 34 out of 37 fecal samples collected from captured deer and the sexes estimated by the present method were perfectly matched with the actual sexes.     

单核苷酸多态性(SNPs)——第三代DNA分子标记

单核苷酸多态性 (SNPs)是指染色体基因组水平上由单个核苷酸变异引起的DNA序列多态性 ,具有分布广、密度高、多态性丰富等特点 ,且易于实现自动化检测 ,被公认为是最新的第 3代DNA分子标记。其在基因作用机理、疾病的研究及动物育种等领域具有广泛的应用前景。本文简要介绍了 6种高通量的SNPs检测技术 :基因芯片、TaqMan、变性高效液相色谱、双色荧光偏振检测入侵、基质辅助激光解吸附电离飞行时间质谱及单碱基延伸标签阵列的原理及其优、缺点。