A History of Equine Embryo Transfer and Related Technologies

The first successful equine embryo transfer was reported in 1972, 21 years after the first reported embryo transfer in cattle. Adaptations of embryo transfer and the various related technologies have generally been more rapid in the equine than in cattle, with the exceptions of superovulation, in vitro fertilization and cryopreservation. Recent progress has been achieved in all three of these areas. This paper presents a time line of various events in the history of equine embryo transfer and related technologies. Approximately 25,000 equine embryo transfers are being conducted per year, worldwide.     

驴源马疱疹病毒8型的分离鉴定与生物信息学分析

本研究旨在分离1株驴源马疱疹病毒8型（Equine herpesvirus type 8,EHV-8）毒株并分析其遗传特征。从山东省聊城地区某驴场采集病驴肺脏组织,经PCR方法鉴定EHV-8的感染情况;对EHV-8感染阳性的肺组织进行研磨,经反复冻融后接种于兔肾细胞（RK-13）细胞中,盲传3代,待出现细胞病变（CPE）时收集细胞,并通过PCR、间接免疫荧光试验、透射电镜等技术对EHV-8进行鉴定。利用PCR扩增获得分离株的ORF70全基因组序列,并进行生物信息学分析。研究结果显示,经PCR鉴定获得EHV-8感染阳性的肺脏组织,病料接种易感细胞RK-13后出现典型CPE,分别收集前3代的细胞培养物,经PCR扩增,均获得与预期大小一致的ORF70基因片段,将分离获得的EHV-8毒株命名为SDLC66,序列上传GenBank,获得登录号：MW816102。经测序和序列比对发现,SDLC66毒株与AHV-3毒株（GenBank登录号：U24184.1）ORF70基因的相似性为99%,与国内的EHV-8 wh毒株（GenBank登录号：JQ343919.1）、国外EHV-8/IR/2015/40（GenBank登录号：MF431614.1）、EHV-8/IR/2003/19（GenBank登录号：MF431611.1）参考毒株的相似性最高,均为99.8%。经遗传进化树分析发现,SDLC66与EHV-8（EHV-8 wh、EHV-8/IR/2015/40和EHV-8/IR/2003/19株）在一个小分支上,亲缘关系最近;与EHV-1型参考毒株（Hong Kong/57/1984、United Kingdom/32/1982、Oxfordshire/206/2013株）序列来源于同一大分支,亲缘关系较近,与EHV-4毒株（91c1和TH20p株）亲缘关系较远。间接免疫荧光试验可见与病毒蛋白特异结合的红色荧光信号;透射电镜观察可见直径约110 nm的圆形病毒粒子,并且核衣壳外有一层亮晕,亮晕外有一层囊膜。说明本试验成功分离得到1株驴源EHV-8毒株,为进一步研究其致病性和致病机制奠定基础。     

Antimicrobial and Antioxidant Activities of Two Medicinal Plants Cuphea aequipetala var. hispida (Cav.) Koehne and Eryngium comosum Delaroche F Against Bacteria Related to Equine Infections

Functional biocompounds beneficial for animals and humans are in Mexican folk herbs. Cuphea and Eryngium species presented antimicrobial potential. Natural antibiotic uses by ethnoveterinary research with medicinal plants in equine infection or digestive diseases need more scientific evidence. Staphylococcus aureus, Escherichia coli, Salmonella enterica serotype Enteritidis are etiological agents in horses responsible for stable infections, abortions, fetal or perinatal deaths, and resistant intrahospital infections. The main objective of the present research was to evaluate the potential of antimicrobial and antioxidant activities of two Mexican medicinal plants Cuphea aequipetala var. hispida (Cav.) Koehne and Eryngium comosum Delaroche F over Listeria monocytogenes ATCC 19115, Staphylococcus sp., E. coli ATCC 25922, and S. enterica serotype Enteritidis ATCC 13076 bacterium reference strains related to equine infections. Determination of total phenol, saponins, antioxidant activity (ABTS), and antimicrobial activity with diffusion-sensitive discs was performed in triplicate. All the strains were sensitive for both extracts except for E. coli strain that was inhibited only by C. aequipetala. Staphylococcus sp. and S. enterica strains were inhibited equally by both extracts. E. comosum extracts tested have shown the highest effect over L. monocytogenes. In summary, antimicrobial activity was similar to the reported activity of Eryngium species extracts with other different solvents. Present extracts are suggested as a potential alternative antibiotic; definitely, more specific equine pathogen inhibition tests are needed in feed additives for horse nutrition research. In conclusion, antimicrobial activities of Cuphea aequipetala var. hispida (Cav.) Koehne and Eryngium comosum Delaroche F over reference strains related to equine infections suggested these medicinal plants as potential antibiotic sources for horse diseases.     

The Effect of Season on the Histologic and Histomorphometric Appearance of the Equine Pituitary Gland

The objective of this study was to determine the effect of season on the histologic and histomorphometric appearance of the normal equine pituitary gland. Pituitary glands were collected at necropsy from 121 horses throughout the year. Plasma was also collected from 59 of these horses before euthanasia. Hematoxylin and eosin stained median sagittal sections of each pituitary were evaluated and histologically graded by three pathologists. Histomorphometric analysis was performed on the same slides. Plasma α-melanocyte stimulating hormone was measured by radioimmunoassay in a subset of horses (n = 59). A total of 118 pituitary glands were included in the study after exclusions were made on the basis of the presence of pars intermedia (PI) adenomas (>5 mm). There was a positive correlation between PI hormone concentration (α-melanocyte stimulating hormone) and PI area. Pituitary gland measurements and grades from samples collected in the fall were compared with those collected in the nonfall months using t-test. The PI area, total pituitary area, and PI/total pituitary ratio were significantly greater in the fall compared with nonfall months (P < .0001, P < .01, P < .0001, respectively). Pituitary grades were also higher in the fall compared with nonfall months (P < .001). There was no seasonal difference in pars distalis or pars nervosa area. The results of this study show that the normal equine pituitary shows seasonal changes in appearance and size. These changes must be considered when using postmortem histologic evaluations in the diagnosis of pituitary pars intermedia dysfunction or for validation of antemortem diagnostic tests.     

The Effects of Feed Form on Consumption Time and Glucose and Insulin Response to a Concentrate Meal in Equine

This study tested the hypothesis that feeding an identically formulated, low sugar and starch concentrate in three forms (5-mm extruded [E], 4-mm pellet [P], and 19-mm oval [O]) would affect consumption rate and glucose or insulin responses, or both. Horses received 1.8 kg treatment feed in a randomized, crossover design, with samples taken every 30 minutes for 6 hours for blood glucose and insulin response. Pearson's correlation compared consumption time, insulin and glucose peak, and time to peak insulin and glucose. The pellet (P) elicited a lower (P = .01) glucose concentration at 2.5 hours than O. The pellet also elicited a lower (P = .03) insulin concentration at 5.5 hours than E and O. There were no differences (P > .05) in area under the curve (AUC) insulin, peak insulin, and time to peak insulin for the three treatments. Average insulin concentration was lower (P = .01) for P versus O. There were no differences (P > .05) in average insulin between P and E, nor between O and E. There were no differences (P > .05) in AUC and peak glucose concentration. Time to peak glucose was longer (P = .04) for P versus E. Average glucose concentration was lower (P = .02) for P versus O. Consumption time was longer (P = .03) for O versus P. There was a positive correlation between consumption time and time to peak insulin (r = 0.46, P = .029). Further research on feeding practices, feed forms, and consumption times that affect glycemic response is necessary.     

2007年华北地区H3N8亚型马流感病毒的分离与鉴定

2007年10月,华北地区某赛马场的马同时发生了以发烧、流水样鼻汁或脓性分泌物、咳嗽等临床症状为主的疾病,疑似马流行性感冒。采集患病赛马的鼻腔分泌物,发病期和发病后14d血清,经鸡胚接种法分离病毒,并用鸡红细胞血凝抑制试验(HI)、神经氨酸酶抑制试验(NI)、病毒回归试验、血清学检测和基因序列分析对分离的病毒进行了系统鉴定。结果表明分离的毒株(A/equine/Huabei/1/2007(H3N8)为马源H3N8亚型马流感病毒,基因型属于美洲分支。我们通过动物回归感染试验建立起分离毒株的实验感染模型。     

The Effects of Nonstructural Carbohydrate Content and Feeding Rate on Glucose and Insulin Response to Meal Feeding in Equine

The following research encompassed two experiments and involved feeding horses two isocaloric diets (diet A and diet B), with an approximate 50% difference in nonstructural carbohydrate (NSC) content. There were three main objectives: first, to test the hypothesis that feeding an approximately 50% lower NSC concentrate feed would cause a lower glucose and insulin response; second, to test the hypothesis that feeding meals equal in NSC content would create similar responses in glucose and insulin dynamics; and finally, to test the hypothesis that the time spent eating is correlated with glucose/insulin response. In experiment 1, in which diet A and diet B were fed at the same rate, the main finding was that feeding a meal lower in NSC resulted in a lower glucose and insulin response to the feed. In experiment 2, in which the effects of feeding diets A and B at a rate to provide 0.3 g/kg body weight (BW) NSC per meal were explored, the main finding was that, although glucose responses were similar, the meal containing more NSC/kg and fed at the lower rate resulted in a substantially lower insulin response. Consumption time also was found to be significantly different between treatments.In conclusion, a low NSC formulation and small meal size appear to be sensible recommendations for horses that may benefit from a low glucose and insulin response to feeding. In addition to NSC content, meal size, and nutrient:calorie ratio, nutrient requirements of the individual horse and the entire nutritional balance of the diet also should be addressed.     

Evaluating the Benefits of Equine Massage Therapy: A Review of the Evidence and Current Practices

Following the lead of human athletic training, equine massage therapy is becoming a more common part of the management of equine athletes and pleasure horses alike. The basic science rationale for massage is supported by research indicating that massage may affect a number of physiologic systems as well as cellular and fascial components of the muscular system. Equine therapeutic massage, or sports massage, employs a number of techniques first developed in humans and has been reported to increase range of motion and stride length, reduce activity of nociceptive pain receptors, and reduce physiologic stress responses. Additional preliminary research indicates that massage therapy also may improve some aspects of exercise recovery. Although important evidence has begun to document the potential benefits of massage therapy for equine athletes, the current review may say less about the true clinical effects of massage therapy than it does about the current state of research in this field. Additional prospective study of massage therapy using sufficient scientific rigor will be necessary to provide veterinarians, trainers, and owners with definitive data and scientifically based confidence in the use of equine massage. In the meantime, the preliminary research, anecdotal positive effects, and case studies indicating potential benefit are not to be ignored; equine massage therapy already plays a valuable practical role in the care and training of many equine athletes.     

Puncture of the Equine Embryonic Capsule and Its Repair In Vivo and In Vitro

Vitrification of embryos >300 µm in diameter requires puncture of the glycoprotein capsule, although the size of the hole compatible with embryo survival is unknown. Forty-five day-7 or -8 embryos were punctured using a 30-µm glass biopsy pipette mounted on a micromanipulator (n = 20) or manually with either an acupuncture needle (∼100-µm diameter -hole; n = 10) or a microneedle with a <1 µm tip to produce a ∼30-µm diameter hole (n = 15) before transferring to recipient mares; further 12 embryos were punctured with either the acupuncture needle or microneedle before being cultured in vitro for 48 hrs (n = 3 per puncture group) or transferred to recipient mares and recovered 48 hrs later (n = 3 per puncture group). No pregnancies resulted from the 10 embryos punctured with the acupuncture needle, whereas 15 of 20 (75%) and 10 of 15 (67%) punctured on the micromanipulator or manually with the microneedle resulted pregnancies. Neither acupunctured nor microneedle-punctured embryos repaired their capsules in vitro. The acupunctured embryos also failed to repair their capsule after 48 hrs in vivo and subsequent uterine flushing yielded numerous capsular vesicles. The microneedle-punctured embryos did repair their capsule in vivo. Puncture with the microneedle opens the way for development of a manual method to vitrify equine embryos.