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.     

Effects of adding a commercial capsaicin fertilizer on the feeding behavior of sika deer (Cervus nippon)

We investigated the inhibitory effect of capsaicin fertilizer on feeding in deer. We tested four captive adult female deer. In Experiment 1, in addition to the treatment (intact) containing only a solid feed (HC), we mixed the fertilizer not containing capsaicin (F) or the capsaicin fertilizer (CF) in the solid feed. In addition, the solid feed was put on a wire net that capsaicin fertilizer was placed 5 cm below (SCF). We investigated their feeding behavior response. In Experiment 2, we changed the amount of substance (fertilizer and capsaicin fertilizer) mixed in the HC. We mixed different amounts (0, 50, 100, and 200 g) of the treatments other than the intact with HC and presented them to the deer, and investigated their feeding behavior response. In Experiment 1, intake in the F and CF decreased (p < .05). In Experiment 2, HC intake was significantly lower in the 100 and 200 g CF (p < .05). However, HC intake relatively increased by the last day in the CF 200 g too. The capsaicin fertilizer decreased the feeding behavior of deer by directly touching the mucous membranes of the deer nose and lips. However, the effects were decreased over time.     

A survey of antimicrobial resistance in Escherichia coli isolated from wild sika deer (Cervus nippon) in Japan

We examined the antimicrobial susceptibility of 848 Escherichia coli isolates from 237 feces samples of wild sika deer (Cervus nippon) captured between 2016 and 2019 in 39 of the 47 prefectures of Japan. Five of the 237 wild sika deer (2.1%) carried E. coli with resistance to at least one antimicrobial, and all the resistant isolates showed resistance to tetracycline. The resistant isolates contained antimicrobial resistance genes that were similar to those in E. coli derived from humans and farm animals. Although wild sika deer are not currently likely to be a source for the transmission of antimicrobial resistance in Japan, they can potentially mediate antimicrobial resistance spread by coming into contact with humans, animals, and their surroundings.     

Population pharmacokinetics of methadone hydrochloride after a single intramuscular administration in adult Japanese sika deer (Cervus nippon nippon)

ObjectiveTo assess the population pharmacokinetics of methadone in deer.Study designProspective non-randomized experimental trial.AnimalsTwelve healthy adult sika deer (nine males and three females).MethodsDeer received intramuscular administration of racemic methadone hydrochloride at 0.5 mg kg⁻¹ or 1 mg kg⁻¹. Plasma methadone and its metabolite 2-Ethylidene-1,5-Dimethyl-3,3-Diphenyl-Pyrolidine (EDDP) concentrations were determined by validated liquid chromatography coupled to tandem mass spectrometry methods, at times 0, 30 minutes, 1, 2, 3, 4, 5, 6, 8, 12 and 24 hours. Population pharmacokinetics analysis was undertaken using a non-linear mixed effects modelling (NONMEM).ResultsA two-compartment linear disposition model best described observed time-concentration profiles of methadone and EDDP. Population parameter estimates of methadone were elimination clearance (17.3 L hour⁻¹), metabolic clearance (34.6 L hour⁻¹), volume of distribution of compartment 1 (216.0 L) and volume of distribution of compartment 2 (384.0 L). Population parameter estimates of EDDP were elimination clearance (121.0 L hour⁻¹), volume of distribution of compartment 3 (1.08 L) and volume of distribution of compartment 4 (499.5 L). The total clearance and total volume of distribution of methadone and EDDP were 51.9 L hour⁻¹, 121.0 L hour ⁻¹, 600.0 L and 500.6 L, respectively. The methadone terminal elimination half-life was 8.19 hours. No adverse effects were observed after methadone administration.Conclusions and Clinical relevanceFollowing intramuscular injection, methadone was characterized by a large total volume of distribution, high systemic clearance and intermediate terminal half-life in sika deer.     

梅花鹿天然Toll样受体9基因克隆与序列分析

Trizol法提取梅花鹿肝脏总RNA,采用ORF两端兼并引物,RT-PCR方法克隆梅花鹿天然Toll样受体9基因（TLR9）并进行系统的生物信息学分析。RT-PCR结果显示,梅花鹿TLR9基因的mRNA长4 043bp,含有800bp左右的5’UTR,完整ORF编码1 081个氨基酸（GenBank序列号为：HQ260632）。同源性分析显示梅花鹿TLR9基因编码区与其他物种高度同源,但5’UTR区存在基因结构的变异。功能结构域分析显示不同物种间TLR9结构域相对保守但也存在细微差别,结构域的差别导致梅花鹿TLR9蛋白胞外区马蹄形弧顶内侧空间结构由人类TLR9蛋白的弧形改变为梅花鹿TLR9蛋白的三角形。进化分析显示TLR9基因分子进化关系与物种间真实进化相一致。     

Seasonal changes in digestibility, passage rate and rumen fermentation of alfalfa hay in sika deer (Cervus nippon) under restricted feeding

To investigate seasonal variations in the digestive functions of sika deer, five female sika deer were provided with an amount of alfalfa hay cubes equivalent to voluntary food intake during winter. We measured the rate at which the food passed through the digestive tract, digestibility and rumen fermentation during the summer (August), autumn (November), winter (February) and spring (May). Total mean retention time in the digestive tract during summer and autumn was numerically longer than that in winter and spring, but the difference did not reach significance. Organic matter and fiber were less digestible in summer and autumn than in winter and spring (P < 0.05), whereas the digestibility of the dry matter tended to vary with the seasons (P < 0.1). Ruminal pH values seasonally changed (P < 0.01), and were the lowest in autumn. The concentration of ruminal ammonia‐nitrogen differed significantly among the seasons (P < 0.05), increasing in winter and decreasing during spring and summer. The numbers of protozoa changed significantly among the seasons (P < 0.05), being higher in autumn than in winter and spring, and intermediate in summer. The concentration of total volatile fatty acids was not seasonally affected, but the molar percentages of propionic acid and butyric acid significantly changed according to season (P < 0.05 and P < 0.01, respectively), and the ratio of acetic to propionic acid tended to change with the seasons (P < 0.1). The results of this study suggested that the digestive functions in sika deer, fed a commercial diet at a restricted level, differed notably among the seasons and these variations might partially be due to environmental effects.     

Comparison of digestibility, passage rate and rumen fermentation between sika deer (Cervus nippon) and cattle fed alfalfa hay cubes

Five female sika deer and three male Holstein cattle were offered alfalfa hay cubes at 2% (deer) and 2.5% (cattle) of bodyweight, respectively. The passage rate through the digestive tract, digestibility and rumen fermentation of the animals were determined. The rate of ruminal passage was higher and the total mean retention time in the digestive tract was shorter in deer than in cattle. In addition, the rate of post‐ruminal passage in deer was lower. The digestibilities of dry matter, organic matter, crude protein and fiber in deer were significantly lower than in cattle (P < 0.05). The concentration of total volatile fatty acids in the rumen was significantly higher in deer than in cattle. The molar percentage of acetic acid was lower and that of valeric acid was higher in deer (P < 0.05). The number of protozoa was somewhat higher in deer. These results suggested that the lower digestibility in deer might be a result of the shorter retention time in the digestive tract.     

梅花鹿朊蛋白基因(PRNP)的克隆及序列分析

根据GenBank中鹿朊蛋白基因序列设计特异性扩增引物，采用PCR方法从中国梅花鹿基因组中扩增得到梅花鹿的朊蛋白基因，采用PCR产物直接测序，并将其克隆到pGEM—TEasy载体中测序进行进一步确认，通过分析表明所克隆的梅花鹿朊蛋白基因的ORF基因片段包含771bp，编码256个氨基酸的前体蛋白，相对分子质量约为28200。与已报道的其他品种鹿朊蛋白基因序列进行对比分析，核苷酸及氨基酸序列的同源性均在98％以上。本试验为进一步研究中国梅花鹿朊蛋白的多态性提供了数据。     

Evaluation of the genetic structure of sika deer (Cervus nippon) in Japan's Kanto and Tanzawa mountain areas,based on microsatellite markers

The browsing habits of sika deer (Cervus nippon) in Japan have caused serious ecological problems. Appropriate management of sika deer populations requires understanding the different genetic structures of local populations. In the present study, we used 10 microsatellite polymorphisms to explore the genetic structures of sika deer populations (162 individuals) living in the Kanto region. The expected heterozygosity of the Tanzawa mountain range population (Group I) was lower than that of the populations in the Kanto mountain areas (Group II). Our results suggest that moderate gene flow has occurred between the sika deer populations in the Kanto mountain areas (Group II), but not to or from the Tanzawa mountain range population (Group I). Also, genetic structure analysis showed that the Tanzawa population was separated from the other populations. This is probably attributable to a genetic bottleneck that developed in the Tanzawa sika deer population in the 1950s. However, we found that the Tanzawa population has since recovered from the bottleneck situation and now exhibits good genetic diversity. Our results show that it is essential to periodically evaluate the genetic structures of deer populations to develop conservation strategies appropriate to the specific structures of individual populations at any given time.     

东北梅花鹿优化育种规划中育种目标的确定

讨论了东北梅花鹿依据经济指标确定育种目标的问题 ,以及在东北梅花鹿育种规划中 ,应如何挑选在综合育种目标中的生产性状及其对应的选择性状。并着重介绍了应用综合育种值作为数量化育种目标的方法。指出在确定以茸用为主的东北梅花鹿育种目标时 ,除了主要考虑茸用性状外 ,还应适当地考虑繁殖和使用寿命等性状 ,同时对于直接影响茸鹿生产效益的直接性状和次级性状不容忽视。     

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.     

Effects of dietary manganese supplementation on nutrient digestibility and production performance in male sika deer (Cervus Nippon)

This study aimed to investigate the effects of manganese (Mn) supplementation on nutrient digestibility and antler growth in male sika deer. Twenty 3‐year‐old male sika deer were divided into four groups (Control, T40, T80 and T120). The control diet contained 22.2 mg Mn/kg. The treatment diets were supplemented with 40, 80 and 120 mg of Mn/kg (provided as Mn methionine). The results showed that the digestibility of dry matter and crude protein in the T80 group was greater than other groups. Higher digestibility of neutral detergent fiber and acid detergent fiber was observed in the T80 group than in the control group. Plasma Mn concentrations in the T80 and T120 groups were higher (P < 0.01) than control and T40 group (P < 0.01). The concentration of Mn in feces was increased with increasing Mn amount. Average daily gain of fresh antler and dry antler of T80 was greater than other groups, especially the control group (P < 0.05). In conclusion, the suitable level of Mn supplementation was found to be 80 mg/kg (total Mn content 103.4 mg/kg dry matter), which significantly increased antler daily gain and feed digestibility in 3‐year‐old sika deer.     

Daily incremental lines in sika deer (Cervus nippon) dentine

This work was designed to observe the dentine incremental lines of the sika deer (Cervus nippon) fawns and to investigate their periodicity using the chronological labeling method with fluorochromes. The incremental lines were observed in decalcified specimens stained by Bodian's silver technique, and the fluorescence-labeled lines were observed in undecalcified and ground specimens. In the silver stained specimens, there were two types of lines, deeply stained thick lines and faintly stained minute regular incremental lines. The intervals and staining intensities of the deeply stained thick lines were very similar to those of the fluorescence-labeled lines in the ground specimens obtained from the same tooth, and hence, it appeared that the both lines were identical. The number of minute incremental lines between the deeply stained thick lines was the same as that of days between the time when each fluorescent labeling injection was made. Therefore, it seemed that each minute incremental line was formed each day. The possibility of age estimation in days using diurnal dentine increments was discussed.     

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.     

梅花鹿鹿茸异位生长研究

为了揭示鹿茸发生机理,阐明生茸区骨膜在鹿茸发生中的作用,选择4只1周岁雄性梅花鹿,在鹿茸发生前,通过外科手术将一侧完整的生茸区骨膜摘除并移植到额骨上。结果,摘除骨膜的生茸区没有发生鹿茸,而在被移植的额骨部位上发生了鹿茸且生长发育正常。证明了生茸区骨膜是鹿茸发生的组织基础,同时为通过异位生茸提高鹿茸产量开辟了新的途径。     

Hematology in sika deer (Cervus nippon yesoensis Heude, 1884)

Blood samples were taken from 78 wild and 21 farmed sika deer (Cervus nippon yesoensis) using ketamine-xylazine sedation during their excited (82 deer) and resting (17 deer) states. Red cell count (RBC), hemoglobin (Hb), packed cell volume (PCV) and mean corpuscular volume (MCV) were significantly higher and mean corpuscular hemoglobin concentration (MCHC) was lower in excited deer than in resting deer. There was no significant difference in total leukocyte count (WBC) between excited and resting wild males, while a marked increase of WBC with neutrophilia was observed in excited wild females. RBC and PCV were significantly higher and MCH was lower in excited males than in excited females. In wild deer, WBC was significantly higher in females than in males, but there was no significant difference in WBC between farmed males and females. Sex differences in the hematological parameters were not observed in fawns (10 months).     

Bcl-2在梅花鹿茸角中的表达

鹿茸角是哺乳动物唯一的失去后还能完全再生的器官,人们对鹿茸角再生的分子机理了解甚少。本试验以梅花鹿为研究对象,通过原位杂交方法对Bcl-2在梅花鹿茸角中的表达进行了研究。结果显示,Bcl-2在梅花鹿茸角表皮层内表达甚微,在茸角真皮层、间充质层及软骨层等处均有表达,但在表达强度上存在一定差异。在真皮层中,Bcl-2在真皮成纤维细胞中有较强的表达,在鹿茸间充质细胞中也有少量Bcl-2的表达;在鹿茸软骨层中,Bcl-2在软骨细胞中的表达量很高,主要表达在增殖区的软骨细胞中。这表明Bcl-2可能在梅花鹿茸角再生过程中起重要的调节作用。     

An occurrence of Salmonella typhimurium infection in sika deer (Cervus nippon)

Seven sika deer (Cervus nippon) in a herd of 30 deer in a park died. Upon examination of three dead deer, Salmonella Typhimurium was isolated from the organs and intestinal contents. Histopathological examination revealed catarrhal enteritis and focal necroses in the liver. Immunohistochemically, Salmonella antigen of O4 was detected in the enteric lesions. The case was diagnosed as S. Typhimurium infection in the sika deer. Because of the importance of Salmonella in public health, fecal and soil samples were continuously collected from the paddock. However, no Salmonella was isolated from any samples collected after medication of the deer and thorough disinfection of the immediate environment.     

基于SRY基因分析梅花鹿的遗传多样性及父系类型

试验旨在研究梅花鹿SRY基因的遗传多样性及其父系类型。选取东北亚种、北海道亚种、本州亚种、指名亚种、屋久岛亚种5个亚种的144个个体,利用试剂盒和传统酚/仿(1∶1)法进行DNA的提取,采用PCR扩增和直接测序法分析梅花鹿的核苷酸多样性(Pi)、单倍型多样性(Hd)、遗传距离及系统进化关系。结果显示,试验所获序列长度为1 613bp,在第47、62、602、1 148、1 569、1 604bp处共检测到6个SNPs多态位点,占核苷酸总数的0.3%,碱基替换以碱基转换为主。通过6个SNPs多态位点确立了6种单倍型:Hap-1、Hap-2、Hap-3、Hap-4、Hap-5和Hap-6,其中Hap-4、Hap-5和Hap-6为新发现单倍型。遗传多样性由高至低依次为:指名亚种、屋久岛亚种、东北亚种、本州亚种、北海道亚种。各亚种间遗传距离最小为北海道亚种与本州亚种的距离(0.000056),最大为东北亚种与指名亚种的距离(0.001733)。基于单倍型利用邻接法构建系统进化树,结果显示,与日本梅花鹿相比,东北亚种与马鹿关系更近,东北亚种存在两大分支,日本梅花鹿各亚种间无明显分支,其他单倍型均是由Hap-3进化而来,单倍型最小跨度网络图与系统进化树一致。结果表明,东北梅花鹿存在两大父系类型,日本梅花鹿存在一个父系类型,单倍型Hap-3是日本梅花鹿的原始单倍型。