Extent of Mycobacterium bovis infection in dairy cattle herds subject to partial culling as determined by an interferon-gamma assay

The interferon-gamma (IFN-γ) assay is employed as a complementary diagnostic test for bovine tuberculosis (BTB) in many countries. To simplify this assay, we established a 96-well plate format using the ESAT-6 and CFP-10 antigens and then employed it to determine the extent of Mycobacterium (M.) bovis infection in dairy herds with a history of BTB outbreaks in a country where only selective culling is practiced. The sensitivity and specificity of this IFN-γ assay were 85.9% and 100%, respectively, based on comparison with the conventional single intradermal tuberculin test (SIDT). The IFN-γ assay was also positive in 30.4% and 36.8% of SIDT-negative animals from herds with recent and remote BTB outbreaks, respectively. Of 14 SIDT-negative, IFN-γ positive cattle, five (35.7%) were culture positive and an additional six were positive based on a polymerase chain reaction-based test for M. bovis. Therefore, the IFN-γ assay has the potential to serve as a specific and sensitive test for M. bovis infection in dairy cattle. Further, the results indicated that a substantial portion of SIDT-negative animals in herds with previous BTB outbreaks were actually infected with M. bovis. Accordingly, the present selective-culling strategy may require modifications to include this more sensitive assay.     

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。     

First detection of adiaspiromycosis in the lungs of a deer

Adiaspiromycosis is a pulmonary infection caused by the soil fungi, Emmonsia crescens and E. parva. It primarily affects small mammals and can range from an asymptomatic condition to fatal disseminated disease. We detected a granuloma containing fungal spherules, which were morphologically consistent with the adiaspores of E. crescens in the lungs of a female Hokkaido sika deer. This is the first reported case of adiaspiromycosis involving a cervid in the world.     

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。     

细胞周期蛋白D1在梅花鹿茸角中的表达

以梅花鹿为研究对象,通过原位杂交方法对细胞周期蛋白D1（cyclin D1）在梅花鹿茸角中的表达进行了研究。结果显示,cyclin D1在梅花鹿茸角表皮层内表达极少,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差异。在真皮层中,cyclin D1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cyclin D1的表达,在鹿茸软骨层中,cyclin D1在软骨细胞中的表达量非常高,主要表达在增殖区的软骨细胞中。结果表明,cyclin D1可能在梅花鹿茸角再生过程中起重要的调控作用。     

Molecular findings and approaches spotlighting Mycobacterium bovis persistence in cattle

Mycobacterium tuberculosis (Mtb) and Mycobacterium bovis (M. bovis) are the etiological agents of human and bovine tuberculosis (TB, bTB) respectively, and share genetic identity over 99% at the whole genome level. Progress has been made towards explaining how mycobacteria and their infected hosts remain in balance without producing clinical symptoms of disease, a phenomenon referred to as latency or persistence, which can be mimicked by certain in vitro conditions. Latency/persistence has mainly been studied using Mtb, where the two-component signalling system, dosRS, has been assigned an instrumental role, and even constitutes the current basis for development of new diagnostic methods and treatment addressing this particular stage of TB. M. bovis conserves homolog genes that in Mtb play a role in human latent TB infection and that, by analogy, would allow it to enter a persistent state in infected cattle; nevertheless, little attention has been paid to this stage in bovine hosts. We suggest that many of the advances acquired through the study of Mtb can and should be taken into consideration by research groups and veterinary professionals dealing with bTB. The study of the infection in bovines, paying particular attention to defining the molecular and cellular markers of a M. bovis persistent infection in cattle, presents great opportunities for the development and trial of new diagnostic tests and vaccines, tools that will surely help in promoting eradication of bTB in high-burden settings.     

梅花鹿增茸综合技术的试验研究

选择５８只成年梅花鹿，随机分为两组。试验组３８只，在精料补充料中添加 １％的鹿用预混料，并于收获头茬茸时，颈部皮下注射“再生茸生长剂”６ｍｌ；对照组 ２０只，不使用预混料和“再生茸生长剂”。结果表明，试验组三杈茸鲜重比对照组提高１３．４９％（Ｐ〈０．０５），再生茸平均单产提高２２１．０５％（Ｐ〈０．０１）。试验组平均纯增经济效益是增加投入的７．５６倍。     

吉林省梅花鹿副结核病、结核病及布鲁菌病血清流行病学调查

采集吉林省具有代表性的4个地区的8个梅花鹿场（四平2个梅花鹿场、通化2个梅花鹿场、长春市2个梅花鹿场、辽源2个梅花鹿场）中630份血清样本作为研究对象,进行血清学（ELISA_方法）检测,了解吉林省梅花鹿副结核病、结核病及布鲁菌病感染现状。上述4个地区均存在梅花鹿副结核病,结核病及布鲁菌病发生和流行,其中副结核病的整体阳性率为18．73％、结核病的整体阳性率为19．21％、布鲁病的整体阳性率为28．42％。吉林省梅花鹿副结核病、结核病及布鲁菌病流行不容乐观,提示广大饲养者及相关部门应该高度重视梅花鹿副结核病的防治工作。     

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.     

Tuberculosis in alpaca (Lama pacos) on a farm in Ireland. 2. Results of an epidemiological investigation

Tuberculosis (TB), due to infection with Mycobacterium bovis was diagnosed in a flock of alpaca in Ireland in 2004. An epidemiological investigation was conducted to identify the risk of TB for farmed alpaca where TB is endemic, the origin of the infection, the potential for alpaca-to-alpaca transmission and appropriate control measures. The investigation focused on the alpaca flock (including the farm, animal movements and breeding, feeding and flock health practice), the disease episode (including animal disease events and subsequent control measures) and TB infection risk in the locality. The TB risk to alpaca is high in areas where infection is endemic in cattle and badgers and where biosecurity is inadequate. It is most likely that the source of infection for the alpaca was a local strain of M. bovis, present in cattle in this area since at least 2001. Genotyping of isolates identified a single variable number tandem repeat (VNTR) profile in both cattle and alpaca in this region. Although a tuberculous badger was also removed from the vicinity, bacterial isolation was not attempted. On this farm, infection in alpaca was probably derived from a common source. Alpaca-to-alpaca transmission seems unlikely. Two broad control strategies were implemented, aimed at the rapid removal of infected (and potentially infectious) animals and the implementation of measures to limit transmission. Tests that proved useful in detecting potentially-infected animals included measurement of the albumin-to-globulin ratio and regular body condition scoring. Skin testing was time consuming and unproductive, and early detection of infected animals remains a challenge. The flock was managed as a series of separate groupings, based on perceived infection risk. No further TB cases have been detected.     

Variable number tandem repeat analysis of Mycobacterium bovis isolates from Gyeonggi-do, Korea

Bovine tuberculosis (TB) is a major zoonosis that''s caused by Mycobacterium bovis (M. bovis). Being able to detect M. bovis is important to control bovine TB. We applied a molecular technique, the variable number tandem repeat (VNTR) typing method, to identify and distinguish the M. bovis isolates from Gyeonggi-do, Korea. From 2003 to 2004, 59 M. bovis clinical strains were isolated from dairy cattle in Gyeonggi-do, Korea, and these cattle had tuberculosis-like lesions. Twenty-four published MIRU-VNTR markers were applied to the M. bovis isolates and ten of them showed allelic diversity. The most discriminatory locus for the M. bovis isolates in Korea was QUB 3336 (h = 0.64). QUB 26 and MIRU 31 also showed high discriminative power (h = 0.35). The allelic diversity by the combination of all VNTR loci was 0.86. Six loci (MIRU 31, ETR-A and QUB-18, -26, -3232, -3336) displayed valuable allelic diversity. Twelve genotypes were identified from the 59 M. bovis isolates that originated from 20 cattle farms that were dispersed throughout the region of Gyenggi-do. Two genotypes [designation index (d.i.) = e, g] showed the highest prevalence (20% of the total farms). For the multiple outbreaks on three farms, two successive outbreaks were caused by the same genotype at two farms. Interestingly, the third outbreak at one farm was caused by both a new genotype and a previous genotype. In conclusion, this study suggests that MIRU-VNTR typing is useful to identify and distinguish the M. bovis isolates from Gyeonggi-do, Korea.     

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。     

Gastrointestinal parasites in an isolated Norwegian population of wild red deer (Cervus elaphus)

Background

Thirteen red deer (Cervus elaphus), culled from the isolated population at the Mongstad Oil Refinery, Norway, were investigated for gastrointestinal helminths. These animals, enclosed by the refinery fence, do not have contact with other ruminants and have a high population density considering the available browsing area (1 km²) within the refinery site (3 km²). The population was estimated to be 110-130 at the time of culling.

Results

The helminth fauna among these sampled red deer was enumerated and species were identified based on morphology. Ostertagia leptospicularis/O. kolchida was detected in 83% [CI 55 - 95%], Spiculopteragia spiculoptera/S. mathevossiani in 92% [CI 65 - 99%] and Trichostrongylus axei in 42%, [CI 19 - 68%] of the abomasa examined. Characterisation of the intestinal parasite fauna revealed Capillaria bovis, Cooperia oncophora, Oesophagostomum venulosum, Trichuris globulosa and tapeworm fragments (presumed anoplocephalids) in seven individuals. Only one calf had an infection with more than one intestinal helminth (tapeworm fragment and Trichuris globulosa). The remaining six deer had single species intestinal infections.No significant age related trends were seen, with the exception of higher intensity of infection of T. axei in yearlings relative to other age classes. Assessment of abomasal parasite burden and body condition revealed no significant trends. In calves, statistically non-significant correlation was seen between increased parasite burden and decreased slaughter weight, whilst the opposite was seen in adults with the heaviest adults exhibiting the higher burdens. Given the small sample size the trends that were seen need further investigation. The parasite burden was aggregated with three adult red deer harbouring 75% of the total abomasal parasite count.

Conclusion

This isolated population was parasitised by a reduced subset of gastrointestinal nematodes typical of this cervid across an extensive geographic range in Eurasia. The intensity and abundance of abomasal nematodes was higher in this isolated population than reported in similar studies of red deer populations across Europe.     

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.     

低蛋白质日粮补充赖氨酸、蛋氨酸对离乳期梅花鹿氮代谢的影响

为研究低蛋白质日粮添加赖氨酸和蛋氨酸对仔鹿氮代谢的影响,采用4×4完全拉丁方试验,选取3月龄、健康、离乳梅花鹿仔鹿4只,体重为(30±0.12)kg(X?±SD),进行消化代谢试验。对照组(Ⅰ组)饲喂16.28%粗蛋白质的高蛋白质日粮,试验组饲喂13.40%粗蛋白质,添加0.23%赖氨酸,并分别添加0(Ⅱ组)、0.06%(Ⅲ组)、0.12%(Ⅳ组)蛋氨酸的低蛋白质日粮。结果表明:1)对照组蛋白质消化率显著高于Ⅱ组(P<0.01)、Ⅲ组和Ⅳ组(P<0.05),脂肪消化率显著低于Ⅱ组(P<0.01),其他营养物质消化率差异不显著(P>0.05)。2)对照组和Ⅳ组蛋氨酸消化率显著高于Ⅱ组和Ⅲ组(P<0.05),对照组其余氨基酸消化率(赖氨酸、胱氨酸除外)显著高于各试验组(P<0.05)。3)对照组吸收氮显著高于Ⅱ组(P<0.01)、Ⅲ组和Ⅳ组(P<0.05),尿氮排放量显著高于各试验组(P<0.05),氮利用率及氮生物学效价低于各试验组(P>0.05)。4)对照组尿嘌呤衍生物总排放量显著大于Ⅱ组(P<0.01)和Ⅲ组(P<0.05),尿囊素显著大于Ⅱ组(P<0.01)、Ⅲ组和Ⅳ组(P<0.05),对照组、Ⅲ组和Ⅳ组黄嘌呤和次黄嘌呤显著高于Ⅱ组(P<0.01)。补充赖氨酸、蛋氨酸可改善低蛋白质日粮仔鹿营养物质消化率利用率,除粗蛋白外其他营养物质消化率与常规蛋白质水平相近,甚至更好,提高氮沉积、氮利用率和生物学效价,改善氮平衡,减少氮排放,降低环境污染。     

精氨酸水平对梅花鹿仔鹿瘤胃发酵参数和菌群结构的影响

为研究精氨酸水平对离乳期梅花鹿瘤胃发酵参数和微生物菌群结构的影响,试验选择3月龄,体重相近的12只健康雄性梅花鹿,随机分为3组,每组4只。仔鹿饲喂蛋白质水平为12.28%的低蛋白质饲粮,精氨酸水平分别为0.80%(A组)、1.08%(B组)、1.26%(C组),试验期50 d。在试验期的最后1 d抽取仔鹿瘤胃液20 mL,一部分用于测定瘤胃发酵参数,一部分用于提取DNA分析瘤胃细菌多样性。结果表明：1)B组氨态氮含量极显著高于C组（P<0.01）;B组乙酸、丁酸、异丁酸含量极显著低于A组（P<0.01）,总挥发性脂肪酸含量显著低于A组（P<0.05）;B组的乙丙比极显著低于A、C组（P<0.01）。2)B组ACE指数显著高于C组（P<0.05）,Chao1指数显著高于A、C组（P<0.05）,B组Simpson指数极显著高于A、C组（P<0.01）,Shannon指数则极显著低于A、C组（P<0.01）,A组细菌群落结构与B、C组差异显著（P<0.05）。3）在门水平上,B组拟杆菌门（Bacteroidetes）相对丰度极显著低于A...     

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.     

不同蛋白质水平对育成早期梅花鹿消化代谢的影响

为探讨不同蛋白质水平配合日粮对育成早期梅花鹿消化代谢的影响,选择4只9月龄雄性梅花鹿,按4×4拉丁方设计,进行消化代谢试验。结果表明,不同蛋白质水平配合日粮(蛋白质水平分别为10.27%、14.26%、15.61%、18.60%)对梅花鹿体重及日增重、干物质采食量及消化率、蛋白质采食量、沉积量、可消化蛋白质量、蛋白质消化率及代谢率、钙磷采食量及消化率、血液总蛋白及白蛋白浓度差异不显著(P>0.05);对血清尿素氮浓度具有显著性差异(P<0.05)。当配合日粮蛋白质水平为15.61%时,蛋白质代谢率为30.64%。蛋白质沉积量y1与蛋白质采食量x1呈正相关,二者之间的回归方程为:y1=0.394x1-27.568,R2=0.882 8,n=8(P<0.05)。     

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.