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Genetic characterization of Japanese native horse breeds by genotyping variants that are associated with phenotypic traits. 通过对与表型特征相关的变体进行基因分型,确定日本本土马匹品种的遗传特征。
Q3 Veterinary Pub Date : 2023-12-01 Epub Date: 2024-01-18 DOI: 10.1294/jes.34.115
Ibuki Saito, Kotono Nakamura, Teruaki Tozaki, Kazuki Hano, Masaki Takasu

Concerns have been raised about the loss of genetic diversity in Japanese native horses because of their declining populations. In this study, we investigated the genetic variation of four genes, myostatin (MSTN), ligand-dependent nuclear receptor corepressor like (LCORL), doublesex and mab-3 related transcription factor 3 (DMRT3), and 5-hydroxytryptamine receptor 1A (HTR1A), which are associated with horse phenotypic traits, in six Japanese horse breeds (Hokkaido, Kiso, Noma, Misaki, Tokara, and Yonaguni). MSTN, LCORL, DMRT3, and HTR1A showed polymorphisms in the Kiso; Hokkaido and Noma; Hokkaido; and Kiso, Tokara, and Yonaguni breeds, respectively. The Misaki did not show polymorphisms in any of the genes. This study may serve as a basis for developing future breeding strategies focusing on traits in Japanese native horses.

由于日本本土马的数量不断减少,人们对其遗传多样性的丧失表示担忧。本研究调查了日本六个马种(北海道马、木曾马、野间马、三崎马、十日原马和米国马)中与马表型特征相关的肌生长蛋白(MSTN)、配体依赖性核受体核心抑制因子(LCORL)、双倍体和 mab-3 相关转录因子 3(DMRT3)以及 5- 羟色胺受体 1A(HTR1A)这四个基因的遗传变异情况。MSTN、LCORL、DMRT3和HTR1A分别在木曾马、北海道马和野间马、北海道马、木曾马、十原马和米国马中出现多态性。而 Misaki 没有在任何基因中出现多态性。这项研究可作为今后制定以日本本土马匹特征为重点的育种策略的基础。
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引用次数: 0
Effects of resveratrol and its analogues on the cell cycle of equine mesenchymal stem/stromal cells. 白藜芦醇及其类似物对马间充质干/基质细胞细胞周期的影响。
Q3 Veterinary Pub Date : 2023-09-01 Epub Date: 2023-09-08 DOI: 10.1294/jes.34.67
Norihisa Tamura, Neda Heidari, Richard G A Faragher, Roger K W Smith, Jayesh Dudhia

Resveratrol (RSV; trans-3,5,4'-trihydroxystilbene) strongly activates sirtuin 1, and it and its analogue V29 enhance the proliferation of mesenchymal stem/stromal cells (MSCs).Although culture medium containing 5-azacytydine and RSV inhibits senescence of adipose tissue-derived MSCs isolated from horses with metabolic syndrome, few studies have reported the effects of RSV on equine bone marrow-derived MSCs (eBMMSCs) isolated from horses without metabolic syndrome. The aim of this study was to investigate the effects of RSV and V29 on the cell cycle of eBMMSCs. Following treatment with 5 µM RSV or 10 µM V29, the cell proliferation capacity of eBMMSCs derived from seven horses was evaluated by EdU (5-ethynyl-2'-deoxyuridine) and Ki-67 antibody assays. Brightfield images of cells and immunofluorescent images of EdU, Ki-67, and DAPI staining were recorded by fluorescence microscopy, and the number of cells positive for each was quantified and compared by Friedman's test at P<0.05. The growth fraction of eBMMSCs was significantly increased by RSV and V29 as measured by the EdU assay (control 28.1% ± 13.8%, V29 31.8% ± 14.6%, RSV 32.0% ± 10.8%; mean ± SD; P<0.05) but not as measured by the Ki-67 antibody assay (control 27.0% ± 11.2%, V29 27.4% ± 10.8%, RSV 27.7% ± 6.8%). RSV and V29 promoted progression of the cell cycle of eBMMSCs into the S phase and may be useful for eBMMSC expansion.

白藜芦醇(RSV;反式-3,5,4’-三羟基二苯乙烯)强烈激活sirtuin 1,其及其类似物V29增强间充质干细胞/基质细胞(MSCs)的增殖。尽管含有5-氮杂胞苷和呼吸道合胞病毒的培养基抑制了从患有代谢综合征的马分离的脂肪组织来源的间充质干细胞的衰老,但很少有研究报道呼吸道合胞菌对从没有代谢综合征马分离的马骨髓来源的间质干细胞(eBMMSC)的影响。本研究的目的是研究RSV和V29对eBMMSCs细胞周期的影响。在用5µM RSV或10µM V29处理后,通过EdU(5-乙炔基-2'-脱氧尿苷)和Ki-67抗体测定来评估源自七匹马的eBMMSC的细胞增殖能力。通过荧光显微镜记录细胞的Brightfield图像和EdU、Ki-67和DAPI染色的免疫荧光图像,并通过P
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引用次数: 1
Construction of an individual identification panel for horses using insertion and deletion markers. 使用插入和删除标记构建马的个体识别面板。
Q3 Veterinary Pub Date : 2023-09-01 Epub Date: 2023-08-09 DOI: 10.1294/jes.34.83
Teruaki Tozaki, Aoi Ohnuma, Mio Kikuchi, Taichiro Ishige, Hironaga Kakoi, Kei-Ichi Hirota, Shun-Ichi Nagata

Individual identification and paternity testing are important for avoiding inbreeding in the management of small populations of wild and domestic animals. In horse racing industries, they are extremely important for identifying and registering individuals and doping control to ensure fair competition. In this study, we constructed an individual identification panel for horses by using insertion and deletion (INDEL) markers. The panel included 39 INDEL markers selected from a whole-genome INDEL database. Genotyping of 89 Thoroughbreds showed polymorphisms with minor allele frequencies (MAFs) of 0.180-0.489 in all markers. The total probability of exclusion for paternity testing, power of discrimination, and probability of identity were 0.9994271269, >0.9999999999, and 0.9999999987, respectively. The panel was applied to 13 trios (sires, dams, and foals), and no contradictions were observed in genetic inheritance among the trios. When this panel was applied to the trios (52 trios) containing false fathers, an average of 7.3 markers excluded parentage relationships. In addition, genomic DNA extracted from the urine of six horses was partially genotyped for 39 markers, and 6-28 markers were successfully genotyped. The newly constructed panel has two advantages: a low marker mutation rate compared with short tandem repeats and a genotyping procedure that is as simple as short tandem repeat typing compared with single nucleotide variant typing. This panel can be applied for individual identification, paternity determination, and urine-sample identification in Thoroughbred horses.

个体鉴定和亲子鉴定对于避免野生动物和家畜小种群管理中的近亲繁殖非常重要。在赛马行业,它们对于识别和登记个人以及兴奋剂控制以确保公平竞争极为重要。在这项研究中,我们使用插入和缺失(INDEL)标记构建了一个马的个体识别小组。该小组包括从全基因组INDEL数据库中选择的39个INDEL标记。89只纯种犬的基因分型显示多态性,所有标记的次要等位基因频率(MAFs)为0.180-0.489。亲子鉴定的总排除概率、辨别能力和同一性概率分别为0.9994271269、>0.9999999999和0.9999999987。该小组应用于13个三胞胎(父系、母系和小马驹),在三胞胎之间的遗传上没有发现矛盾。当这个小组应用于包含假父亲的三人组(52个三人组)时,平均有7.3个标记排除了父母关系。此外,从6匹马尿液中提取的基因组DNA进行了39个标记的部分基因分型,6-28个标记成功进行了基因分型。新构建的小组有两个优点:与短串联重复序列相比,标记突变率低,与单核苷酸变体分型相比,基因分型程序与短串联反复序列分型一样简单。该面板可用于纯种马的个体鉴定、亲子鉴定和尿样鉴定。
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引用次数: 0
Association of the severity of colic in horses with oxidative stress biomarkers, acute-phase proteins, and certain trace elements. 马绞痛的严重程度与氧化应激生物标志物、急性期蛋白质和某些微量元素的关系。
Q3 Veterinary Pub Date : 2023-09-01 Epub Date: 2023-09-08 DOI: 10.1294/jes.34.73
Ali Hajimohammadi, Mohsen Ghane, Masoud Ghari Tehrani, Benyamin Paravar, Ahmadreza Mirzaei, SeyedAmin Razavi, Mohammad Nikzad

Sixty-one horses were enrolled in this study and divided into 3 different groups according to their severity of colic (heart rate, oral mucous membrane color, and abdominal distention): a strangulating colic (SC) group (n=21), non-strangulating colic (NC) group (n=20), and control group (n=20) consisting of randomly selected normal horses without signs of colic. The serum concentrations of haptoglobin, tumor necrosis factor-α (TNFα), nitric oxide (NO), malondialdehyde (MDA), zinc, iron, and copper were evaluated in all horses. The average concentration of TNFα in the SC group was higher than that in the control group (P<0.001). The TNFα concentration was higher in the NC group compared with the control group (P<0.001). Furthermore, the average concentration of TNFα tended to be higher in the SC group compared with the NC group (P=0.052). The average concentration of haptoglobin in the SC group was higher than that in the control group (P<0.001). The average concentration of NO was higher in the SC group compared with the NC group. (P=0.016) The average concentration of MDA was higher in the SC group compared with the control group (P=0.042). Furthermore, the concentration of MDA was higher in the SC group compared with the NC group (P=0.048). TNFα in horses with signs of colic may be a reliable indicator of prognosis and the severity of clinical signs. The haptoglobin concentration may be a useful marker in cases where animals are referred to clinicians a few days after the onset of colic. The concentrations of MDA and NO should be interpreted with caution.

61匹马被纳入本研究,并根据绞痛的严重程度(心率、口腔粘膜颜色和腹胀)分为3组:绞窄性绞痛(SC)组(n=21)、非绞窄性结肠(NC)组(n=20)和对照组(n=20),对照组由随机选择的没有绞痛迹象的正常马组成。评估所有马的血清触珠蛋白、肿瘤坏死因子-α(TNFα)、一氧化氮(NO)、丙二醛(MDA)、锌、铁和铜的浓度。SC组TNFα平均浓度高于对照组(P
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引用次数: 0
Incidence of surgical site infection after internal fixation of the first phalangeal bone and the third metacarpal/metatarsal bone fractures in Thoroughbred racehorses. 纯种赛马第一指骨和第三掌骨/跖骨骨折内固定后手术部位感染的发生率。
Q3 Veterinary Pub Date : 2023-09-01 Epub Date: 2023-09-08 DOI: 10.1294/jes.34.61
Hiroshi Mita, Taisuke Kuroda, Hidekazu Niwa, Norihisa Tamura, Kentaro Fukuda, Minoru Ohta

Surgical site infection (SSI) is one of the major complications of equine fracture surgery. The purpose of this study was to investigate the incidence of and risk factors for SSI after internal fixation of the first phalangeal bone (P1) and the third metacarpal/metatarsal bone (MC3/MT3) fractures in Thoroughbred racehorses. Between 2011 and 2020, 451 cases underwent surgery with screws or a locking compression plate (LCP) for sagittal fractures of P1 or condylar fractures of MC3/MT3. Overall, 2.9% (13/451) of the cases developed an SSI. The incidence was significantly higher in plate fixation (21.4%) than in screw fixation (2.3%). There was no significant association with other variables, such as sex, age, number of screws, experience of surgeon, or prophylactic antimicrobials. The median duration of hospitalization for screw fixation was 14 days without an SSI and 20 days with an SSI, and those for plate fixation were 26 and 25-88 days, respectively, indicating that the development of SSI prolongs the duration of hospitalization. On the other hand, there were no significant differences in discharge and race resumption rates between cases with and without an SSI. These data indicate that the incidence of SSI in this study was low and that it was higher following plate fixation than screw fixation.

手术部位感染是马骨折手术的主要并发症之一。本研究的目的是调查纯种赛马第一指骨(P1)和第三掌骨/跖骨(MC3/MT3)骨折内固定后SSI的发生率和危险因素。2011年至2020年间,451例患者接受了螺钉或无导线心脏起搏器(LCP)手术治疗P1矢状骨折或MC3/MT3髁突骨折。总体而言,2.9%(13/451)的病例出现SSI。钢板内固定的发生率(21.4%)明显高于螺钉内固定(2.3%)。与其他变量(如性别、年龄、螺钉数量、外科医生经验或预防性抗菌药物)没有显著关联。螺钉固定的中位住院时间在没有SSI的情况下为14天,有SSI的患者为20天,钢板固定的患者分别为26天和25-88天,这表明SSI的发展延长了住院时间。另一方面,有SSI和没有SSI的病例在出院率和比赛恢复率方面没有显著差异。这些数据表明,本研究中SSI的发生率较低,钢板内固定后的发生率高于螺钉内固定。
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引用次数: 0
A case of equine piroplasmosis in the Tokyo 2020 Olympic Games. 2020年东京奥运会上的一例马梨状体病。
Q3 Veterinary Pub Date : 2023-09-01 Epub Date: 2023-09-08 DOI: 10.1294/jes.34.93
Hiroko Aida, Jonathan H Foreman, Akihiro Ochi, Yoshimasa Takizawa, Takashi Yamanaka

Equine piroplasmosis is an infectious disease caused by Babesia caballi and Theileria equi. A competition horse that had been imported to the Equestrian Park for the Tokyo 2020 Olympic Games and had a fever over 40°C and severe anemia was diagnosed with equine piroplasmosis by blood smear and direct polymerase chain reaction (PCR) tests for Theileria equi. Treatment with protozoan anthelmintics and whole blood transfusion diminished the fever, improved the anemia, and allowed the horse to return home safely. Preparation for routine cases of this infection should include the development of a system that allows accurate and prompt international dissemination of information and implementation of quarantine measures.

马螺旋体病是由马巴贝虫和马泰勒虫引起的一种传染病。一匹为2020年东京奥运会进口到马术公园的比赛马发烧超过40°C,严重贫血,通过血液涂片和马泰勒氏体直接聚合酶链式反应(PCR)检测,被诊断为马梨形体病。使用原生动物驱虫剂和全血输注治疗可以减轻发烧,改善贫血,并使马安全回家。这种感染的常规病例的准备工作应包括开发一个系统,以便在国际上准确、及时地传播信息并实施隔离措施。
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引用次数: 0
Investigation of optimal procedures for storage and use of plasma samples suitable for gene doping tests. 适合基因兴奋剂检测的血浆样品的最佳储存和使用程序的研究。
Q3 Veterinary Pub Date : 2023-06-01 DOI: 10.1294/jes.34.21
Teruaki Tozaki, Aoi Ohnuma, Mio Kikuchi, Taichiro Ishige, Hironaga Kakoi, Kei-Ichi Hirota, Yuji Takahashi, Shun-Ichi Nagata

Gene doping, which is prohibited in horseracing and equestrian sports, can be performed by introducing exogenous genes, known as transgenes, into the bodies of postnatal animals. To detect exogenous genes, a method utilizing quantitative polymerase chain reaction (qPCR) with a hydrolysis probe was developed to test whole blood and plasma samples, thereby protecting the fairness of competition and the rights of stakeholders in horseracing and equestrian sports. Therefore, we aimed to develop sample storage methods suitable for A and B samples in gene doping tests using blood. For sample A, sufficient qPCR detection was demonstrated after refrigeration for 1 to 2 weeks post collection. For sample B, the following procedures were confirmed to be suitable for storage: 1) centrifugation after sample receipt, 2) frozen storage, 3) natural thawing at room temperature, and 4) centrifugation without mixing blood cell components. Our results indicated that long-term cryopreservation yielded good plasma components from frozen blood samples even though it destroyed blood cells, indicating its applicability to the gene doping test using sample B, which can be stored for later use. Sample storage procedures are as important as detection methods in doping tests. Therefore, the series of procedures that we evaluated in this study will contribute to the efficient performance of gene doping tests through qPCR using blood samples.

在赛马和马术运动中被禁止的基因兴奋剂可以通过将外源性基因(即转基因基因)引入产后动物体内来实现。为了检测外源基因,建立了一种利用酶解探针的定量聚合酶链反应(qPCR)检测全血和血浆样本的方法,从而保护赛马和马术运动的公平竞争和利益相关者的权利。因此,我们的目标是开发适用于血液基因兴奋剂检测中A和B样本的样本存储方法。对于A样品,在收集后冷藏1至2周后证明有足够的qPCR检测。对于B样品,确认以下步骤适合保存:1)样品接收后离心,2)冷冻保存,3)室温自然解冻,4)不混合血细胞成分离心。我们的研究结果表明,长期冷冻保存可以从冷冻血液样本中获得良好的血浆成分,尽管它会破坏血细胞,这表明它适用于使用样本B的基因兴奋剂测试,可以保存以备以后使用。在兴奋剂检测中,样品储存程序与检测方法同样重要。因此,我们在本研究中评估的一系列程序将有助于通过使用血液样本的qPCR进行基因兴奋剂检测的高效性能。
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引用次数: 1
Immunohistochemical markers for equine granulosa cell tumors: a pilot study. 马颗粒细胞肿瘤的免疫组织化学标记物:一项初步研究。
Q3 Veterinary Pub Date : 2023-06-01 DOI: 10.1294/jes.34.37
Aurelia Dolin, Peter Schweiger, Martin Waselau, Monika Egerbacher, Ingrid Walter

Sex cord-stromal tumors (SCSTs), generally referred to as granulosa cell tumors (GCTs) or granulosa-theca cell tumors (GTCTs) in equids, show complex compositions and variable numbers of hormone-producing cells. These tumors can be difficult to diagnose, especially in early stages. Therefore, we tested a panel of antibodies for vimentin, smooth muscle actin, laminin, Ki-67, E-cadherin, calretinin, moesin, p-ezrin, AMH, and aromatase, markers used for tumor composition and classification, progression, and prognosis in human SCSTs, on an exemplary grapefruit-size equine GCT within the left ovary of a 13-year-old mare with stallion-like behavior and elevated testosterone levels in comparison with normal ovarian tissue. The tumor showed a low proliferation rate and prominent moesin and p-ezrin staining in granulosa cells. E-cadherin, calretinin, aromatase, and AMH are suggested to be potential markers for different cell components of equine SCSTs that can support tumor diagnosis and classification.

性索间质肿瘤(SCSTs),通常被称为马科动物的颗粒细胞瘤(gct)或颗粒-膜细胞瘤(GTCTs),具有复杂的组成和数量可变的激素分泌细胞。这些肿瘤很难诊断,尤其是在早期阶段。因此,我们在一匹13岁母马的左卵巢内的葡萄柚大小的马GCT上测试了一组抗体,包括vimentin、平滑肌肌动蛋白、层粘连蛋白、Ki-67、e -钙粘蛋白、calretinin、moesin、p-ezrin、AMH和芳香酶,这些标记物用于人类SCSTs的肿瘤组成、分类、进展和预后,该母马的行为与种马相似,睾丸激素水平高于正常卵巢组织。肿瘤增殖率低,颗粒细胞moesin和p-ezrin染色明显。e -钙粘蛋白、calretinin、芳香化酶和AMH被认为是马SCSTs不同细胞成分的潜在标记物,可以支持肿瘤的诊断和分类。
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引用次数: 0
Efficiency of round bale feeders: comparison of Tombstone versus Hay Saver. 圆捆喂料机的效率:墓碑与干草节省器的比较。
Q3 Veterinary Pub Date : 2023-06-01 DOI: 10.1294/jes.34.51
Kendall A Hyde, Annabelle Altman, Robin Banasek, Melba O Gastal, Eduardo L Gastal

Hay wastage when feeding round bales due to contamination, deterioration, and animal refusal can accrue large financial losses for farmers. The present study investigated the efficiency of the conventional Tombstone-style feeder system compared to the Hay Saver feeder system to reduce hay wastage in feeding round hay bales. Mares were distributed equally into two groups, Tombstone and Hay Saver, and fed six bales per group over 48 days. Hay wastage was collected daily, dried, and weighed, while the mares were weighed weekly. Overall, the Hay Saver feeder showed less hay wastage, higher mean mare weight, and higher consumption rate per horse. The results of this study indicated that the Hay Saver feeder system had higher efficiency compared to the Tombstone feeder system.

饲养圆捆干草时由于污染、变质和动物拒绝而造成的干草浪费会给农民带来巨大的经济损失。本研究调查了传统墓碑式给料系统与干草节育器给料系统的效率,以减少喂圆草包时的干草浪费。将母马平均分为墓碑组和干草拯救组,每组饲喂6包,共饲喂48 d。每天收集干草,晒干并称重,而母马每周称重。总体而言,干草节省饲料显示出较少的干草浪费,更高的平均母马体重,和更高的每匹马消耗率。本研究结果表明,干草节省给料系统比墓碑给料系统具有更高的效率。
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引用次数: 0
Monitoring the positive conversion of anti-erythrocyte antibodies in blood transfusion donor horses. 输血供马抗红细胞抗体阳性转化的监测。
Q3 Veterinary Pub Date : 2023-06-01 DOI: 10.1294/jes.34.47
Hironaga Kakoi, Mio Kikuchi, Taichiro Ishige, Yuko Hirosawa, Shoko Tanaka, Shun-Ichi Nagata

To confirm the positive conversion of antibodies against erythrocyte antigens in horses, possible blood transfusion donor horses selected from draft horse populations were periodically monitored with an indirect antiglobulin (Coombs) test for approximately 3 years. In this study, 19 horses (16 females and 3 males) were investigated, and five mares showed alloantibodies during the monitoring period. Four mares were typically pregnant when positive conversion was detected, whereas no particular cause of conversion could be observed for one mare based on its clinical records. In the analyzed horses, most positive conversions were possibly due to pregnancy, as conversion occurred more often during this period than after parturition. Pregnancy is considered a key event for positive conversion. Additionally, in cases in which unknown causative sensitization is confirmed, continuous monitoring with a test to detect antibodies should be performed, even if the possible donor is selected and maintained.

为了确认马体内抗红细胞抗原抗体的阳性转化,从役马群中选择可能输血的供体马,定期进行间接抗球蛋白(Coombs)检测,监测时间约为3年。本研究对19匹马(母马16匹,公马3匹)进行了调查,其中5匹马在监测期间出现同种异体抗体。当检测到阳性转化时,四匹母马通常怀孕,而根据其临床记录,一匹母马没有观察到特定的转化原因。在分析的马中,大多数积极的转换可能是由于怀孕,因为转换在此期间比分娩后更频繁。怀孕被认为是积极转变的关键事件。此外,在确认未知致敏的病例中,即使选择并维持可能的供体,也应通过检测抗体的试验进行持续监测。
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引用次数: 0
期刊
Journal of Equine Science
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