Monitoring of genetic diversity in the endangered Martina Franca donkey population

The Martina Franca (MF) donkey, an ancient native breed of Apulia, was mostly famous for mule production. The breed was at serious risk of extinction in the 1980s following the decrease in demand for draft animals because they were increasingly replaced by agricultural machinery. Much has been done in the last few decades to safeguard the existing donkey breeds, but the situation remains critical. Successful implementation of conservation measures includes an evaluation of the present degree of breed endangerment, so the aim of this work was to analyze the demographic and genetic parameters of this breed to suggest effective conservation strategies. With a current breed register counting less than 500 recorded animals, the pedigree data set included 1,658 MF donkeys born between 1929 and 2006. Analyses were carried out on the whole data set as well as on a smaller one consisting of 422 living animals. Demographic and genetic variability parameters were evaluated using the ENDOG (v4.6) software. The pedigree completeness level was evaluated as well as the generation length, which was calculated for each of the 4 gametic pathways. This information was obtained from animal birth date records together with those of their fathers and mothers. The effective number of founders (f(e)), the effective number of ancestors (f(a)), the founder genome (f(g)), individual inbreeding (F), average relatedness (AR), and the rate of inbreeding per generation were analyzed to describe the genetic variability of the population. Because pedigree depth and completeness were appropriate, especially regarding the current population, the parameters defining genetic variability, namely, f(e), f(a), f(g), F, and AR, could be reliably estimated. Analysis of these parameters highlighted the endangerment status of the MF donkey. Our special concern was with the increased percentage of males and females exhibiting increased AR values. Moreover, the effective size of the current population, 48.08, is slightly less than the range of the minimum effective size, and the rates of inbreeding per generation found in the current MF population exceed the maximum recommended level of 1%. Such a scenario heightens concerns over the endangered status of the MF breed and calls for proper conservation measures and breeding strategies, such as selecting individuals for mating when relationships are below 12.5%.     

Platelet-Rich Plasma as an Adjunctive Therapy for the Management of a Severe Chronic Distal Limb Wound in a Foal

Distal limb wounds are frequent in horses after traumatic events. The use of platelet-rich plasma (PRP) represents a simple method of treating wounds in equines. A case of a chronic severely contaminated distal limb wound in an 8-month-old foal is presented. The patient was managed with the combination of standard wound therapy (debridement and bandages), surgery, and PRP obtained by a manual tube protocol. No complications were observed with the PRP treatment, and the foal was fully recovered 2 months later. The results from this case report indicate that PRP and its by-products (platelet-poor plasma) could be used as an adjunctive treatment in severe distal limb wounds in horses. A clinical controlled study should be performed to test this hypothesis.     

Intraoperative Analgesic Effect of Intrafunicular Lidocaine Injection During Orchiectomy in Isoflurane-Anesthetized Martina Franca Donkeys

This study aimed to evaluate the analgesic effect of intrafunicular lidocaine during orchiectomy in isoflurane-anesthetized donkeys. For this purpose, 10 adult healthy donkeys were chosen from Martina Franca donkey population. Each donkey underwent two surgical procedures of monolateral orchiectomy under general anesthesia. Starting isoflurane vaporizer setting was 1.5% to maintain a light plane of anesthesia. Regional anesthesia was performed by injecting 10 mL of 2% lidocaine plus adrenalin or an equivalent volume of saline solution into the spermatic cord. According to what was injected into the funiculus before the surgical procedure, each donkey was once assigned to the group L (lidocaine) and once to the group S (saline). End-tidal isoflurane and standard physiological parameters were measured. Compared with groups, monolateral orchiectomy increased mean heart rate in group S; during surgical procedure, the end-tidal isoflurane was significantly lower in group L. No differences were found regarding mean arterial blood pressure, respiratory rate, recovery quality, and metabolic parameters between groups during procedures. In isoflurane-anesthetized donkeys, intrafunicular injection of lidocaine before castration appears to decrease intraoperative nociception and significantly reduces the concentration of the volatile agent to obtain a sufficient surgical anesthesia.     

Pedigree information reveals moderate to high levels of inbreeding and a weak population structure in the endangered Catalonian donkey breed

The Catalonian donkey is one of the most endangered donkey breeds in the world. At present, five main subpopulations exist: AFRAC, which consists of many genetically connected Catalonian localities; Berga, which consists of a single herd located also in Catalunya but under private management; and three minor non‐Catalonian subpopulations (Huesca, Sevilla and Toledo). In this study, we analysed the pedigree information of the Catalonian donkey herdbook to assess the genetic diversity and population structure of the breed. We found that the Catalonian donkey has suffered an important loss of genetic diversity and moderate to high increases of inbreeding because of the abuse of a few individuals in matings. This scenario is mainly characterized by the fact that both the effective number of founders and ancestors for the whole population was 70.6 and 27, respectively, while the equivalent number of founders was 146.5 and the number of ancestors explaining overall genetic variability was 93. In addition, only 14% of animals born between the 1960s and 1970s were significantly represented in the pedigree. Our results also show that subpopulations where breeders exchanged reproductive individuals had low levels of inbreeding and average relatedness. One subpopulation, Berga, was reproductively isolated and showed high levels of inbreeding (F = 7.22%), with average relatedness (AR = 6.61%) playing an important role in increasing the values of these coefficients in the whole pedigree. Using genealogical F‐statistics we have found little evidence of population structuring (F_ST = 0.0083) with major genetic differences among non‐Catalonian subpopulations.     

Periosteal transection of the proximal phalanx in foals with angular limb deformities of the metacarpo/metatarsophalangeal area

Bilateral angular limb deformities of the metacarpo/metatarsophalangeal regions in 2 foals are discussed. Periosteal transection was used to correct the deformity in both foals. In one foal, only the right forelimb was treated because the deformity in the left forelimb did not appear to warrant surgery. Subsequently, an angular limb deformity, which could have been prevented, developed in the left forelimb. A third foal developed a deformity in the proximal phalanx after periosteal transection of the distal third metatarsal bone.     

Femoral fracture repair complicated by vascular injury in a foal

A comminuted, mid-diaphyseal femoral fracture was diagnosed radiographically in a 4-month-old Quarter Horse colt. Disruption of the distal blood supply was suspected, as evidenced by coolness and diminished pulses of the distal portion of the limb. The fracture was repaired by compression plating but the foal's condition continued to deteriorate. A femoral arteriogram of the affected limb was obtained. Positive contrast agent was visible only as far as the mid-shaft of the femur. The foal was euthanatized and the postmortem examination revealed a transected popliteal artery accounting for ischemic necrosis of the distal limb.     

Metastatic Tumor in Pregnancy: Placental Germ Cell Tumor With Metastasis to the Foal

A placental mass was observed in an otherwise-healthy Quarter Horse mare; subsequently, the mare's 52-day-old foal was examined because of hind limb ataxia, urinary incontinence, and raised lesions of the distal limbs. Clinical and biochemical findings were supportive of liver disease and lumbosacral injury. Ultrasonographic evaluation of the abdomen suggested a liver mass, which was confirmed with computed tomography (CT) and determined to be neoplastic via histopathologic evaluation of a liver biopsy sample. Initial histopathology suggested a germ cell tumor. Regions of lysis affecting both femurs and third metacarpal bones and a complete oblique sagittal fracture through the body of the first sacral vertebra were present on CT. Supportive care was provided until CT confirmation of diffuse hepatic neoplasia and vertebral fracture. Necropsy revealed a large multinodular mass within the liver and a pathologic fracture of the first sacral vertebral body. Histopathologic comparison between neoplastic cells examined from the placenta and foal confirmed metastases of a germ cell tumor. Placental tumors are rare in all species; however, when noted in horses, clinicians should be aware of the possibility of metastatic spread to the foal.     

Molecular and cytogenetic paternity testing of a male offspring of a hinny

An alleged male foal of a female mule, whose sire and grandparents were unknown, was identified for its pedigree. Parentage testing was conducted by comparing polymorphism of 12 microsatellite DNA sites and mitochondrial D‐loop sequences of the male foal and the female mule. Both the sequence analysis of species‐specific DNA fragments and a cytogenetic analysis were performed to identify the species of the foal and its parents. The results showed that the alleged female mule is actually a hinny, and the male foal, which possesses 62 chromosomes, qualifies as an offspring of the female hinny and a jack donkey.     

湖北利川马的现状分析及对策研究

对利川马的品种资源近几十年来的资料和调查的结果进行分析。用地方品种资源受威胁的程度和群体的有效大小等指标分析了利川马资源的受威胁程度。结果表明：利川马存栏数量在上世纪70年代以后呈逐渐下降的趋势；利川马的有效群体大小正在逐渐下降，处于潜在威胁中，也正接近最低威胁；整体上看，体尺、体重近几年不断提高．利川马产驹教有下降的趋势。     

Congenital bilateral aplasia of the metacarpophalangeal joints in a foal

A 2‐day‐old Warmblood filly was presented for examination of an angular limb deformity of the left front limb and an upright conformation of both metacarpophalangeal joints. Radiological examination revealed bilateral absence of the metacarpophalangeal joint space with fusion of the third metacarpal bone and first phalanx (synostosis). No treatment was undertaken. The filly was readmitted to the clinic 10 weeks later for bilateral front limb lameness. On radiological examination, the synostosis of the front metacarpophalangeal joints was still present. Physitis of the distal growth plate of the right third metacarpal bone and proximal growth plate of the right proximal phalanx, and an avulsion fracture of the palmaromedial and proximal aspect of the left middle phalanx, with a cystic like lesion on the medial aspect of distal first phalanx and proximal middle phalanx were diagnosed. Given the poor prognosis, the foal was subjected to euthanasia. Post mortem examination confirmed the absence of the metacarpophalangeal joint space with a trabecular bony union between the third metacarpal bone and the first phalanx. A rudimentary joint capsule was present at the level of the absent joints as well as a small zone of articular cartilage, which invaginated over a short distance into the dorsal trabecular bone on the right front limb. On the medial aspect of the left proximal interphalangeal joint, a focal defect of articular cartilage with exposure of subchondral bone was observed. This is the first case report of a foal born with congenital aplasia of both metacarpophalangeal joints. Congenital malformations should be considered as differential diagnosis in lame foals or foals born with angular or flexural limb deformities.     

Active tension‐extension splints: A novel technique for management of congenital flexural deformities affecting the distal limb in the foal

A variety of methods are described for managing distal limb flexural deformities in the foal, including intravenous oxytetracycline and splint or cast use. This case series describes a novel technique that creates an ‘active tension‐extension splint’ by wiring the toe into a custom‐made fibreglass splint and therefore into active extension. A dorsal fibreglass splint is made by halving a cast that is set around the affected leg with padding underneath it, so that it is sculpted to a more appropriate anatomical shape. Cerclage wire is placed through the toe and the dorsal aspect of the splint, then tightened to pull the limb into active extension. Foals with distal limb flexural deformities that were treated in this way were followed up by examination of hospital records and telephone questionnaire. Records were examined for 13 foals treated between 2004 and 2010. One foal developed septic osteitis of the distal phalanx due to suspected laminar penetration; other post operative complications seen were bandage sores and minor cosmetic scarring. Out of 10 foals where follow‐up by questionnaire was available, 8 had complete resolution of their deformity following active tension‐extension splinting, one required inferior check ligament desmotomy for complete correction and one had carpal flexural deformities that did not resolve. All that survived to adulthood are sound and have achieved their intended purpose. This previously unpublished technique using a wire through the toe to create an active tension‐extension splint has a high success rate for correction of congenital flexural deformities affecting the distal interphalangeal and metacarpo‐/metatarsophalangeal joints in the foal. The majority of post operative complications are minor and easily managed. This is a simple technique that can improve the management of neonatal distal limb deformities both in a hospital situation and for equine practitioners in the field.     

Clinical Updates II: Topics of Interest Pertaining to the Distal Limb

Regional limb perfusion is used to treat infections of the distal limb and can be performed using a peripheral vein or the intraosseous route; the peripheral vein is the preferred technique. Other distal limb entities of clinical importance include foot conformation problems, bruising, heel pain, navicular disease, degenerative joint disease, tendonitis and desmitis. Specific clinical problems of the distal limb seen in the foal and aged horse are frequently managed with good foot care.     

Closing wedge ostectomy and medial distal tibia plate application for treatment of severe bilateral angular limb deformities in the hindlimbs of an 8-month-old foal

This case report describes the evaluation, surgical approach and outcome of an 8-month-old foal with severe angular limb deformities of both metatarsophalangeal joints (MTPJ). Radiography and computed tomography (CT) were used to diagnose a windswept deformity of the hindlimbs with associated bilateral physitis. The orthopaedic surgery was planned and rehearsed using three-dimensional (3D) bone models. A bilateral closing wedge ostectomy was performed on the distal MtIII, and 3.5 mm LCP^® medial distal tibia plates were used bilaterally for osteosynthesis. Even though convalescence was prolonged and complicated by partial unilateral implant failure and osteoarthritis (OA), reduction of the deformities was successful and resulted in a sound foal.     

Computed tomographic diagnosis of a fracture of the floor of the pelvis in a Thoroughbred foal

This case report describes the clinical and diagnostic imaging findings, treatment and outcomes of a 15-day-old Thoroughbred foal referred for the evaluation of severe left hindlimb lameness of 8 days' duration. Radiographic examination of the left hindlimb and transcutaneous ultrasonographic examination of the pelvis were unremarkable. A computed tomographic (CT) examination was performed to investigate the proximal aspect of the limb and the pelvis; CT scans of the thorax were also acquired. In this case, CT images allowed detection of a closed, slightly displaced, oblique wedge-like fracture of the left caudal ramus of the pubis and medial ramus of the ischium, allowing an accurate diagnosis to guide appropriate treatment and prognostication. The foal was treated with anti-inflammatory drugs and box-rest. A follow-up by telephone interview with the trainer 3 years later revealed that the horse is in full athletic activity without any residual gait alteration related to the pelvic fracture.     

Management of hydrops amnion in a mare resulting in birth of a live foal

CASE DESCRIPTION: A 19-year-old Thoroughbred mare was evaluated at 265 days of gestation with a markedly distended abdomen and edema of the ventral portion of the abdomen. CLINICAL FINDINGS: The uterus was distended over the pelvic rim, making transrectal palpation of the fetus impossible. Transabdominal ultrasonography revealed excessive amounts of fetal fluid. Results of analysis of fluid obtained via amnio- and allantocentesis confirmed that the amniotic cavity was large. TREATMENT AND OUTCOME: The mare was monitored for signs of weakness of the prepubic tendon and abdominal wall. The fetus and placenta were monitored for signs of stress and pending abortion. Flunixin meglumine and altrenogest were administered to the mare. Parturition was attended and occurred at 321 days' gestation. Postpartum complications in the mare included hypovolemic shock and cardiac arrhythmias. Both conditions were treated, and the mare recovered. The foal was considered small, had bilateral angular limb deformities, and was unable to nurse. The foal was given plasma for failure of passive transfer of immunity. Ten months later, the foal underwent procedures to correct limb deformities. CONCLUSIONS AND CLINICAL RELEVANCE: Hydrops conditions are rare in horses, with hydrops allantois occurring more frequently than hydrops amnion; reportedly result in fetal or neonatal death; and may result in death of or injury to the mare. Close monitoring of maternal and fetal health in combination with supportive treatment of the mare can result in the safe progression of a hydrops pregnancy and the birth of a live foal.     

Use of intralesional cisplatin to successfully treat distal limb haemangiosarcoma in a foal

A 4-month old, 200 kg, grey warmblood colt presented for a firm, non painful mass on the distal medial aspect of the left third metatarsus. Excisional biopsy revealed a diagnosis of haemangiosarcoma. Equine haemangiosarcoma is uncommon and only limited reports of successful treatment are available. The prognosis for survival is therefore considered to be poor. After two separate incidences of recurrence with incomplete excision of the tumour, intralesional treatment with cisplatin without excision or debulking was performed on three separate occasions. Intralesional cisplatin injection was performed at monthly intervals for three treatments. Four years post treatment with cisplatin, the horse remained in remission. This case report describes the diagnostic and treatment challenges for successful treatment of a primary haemangiosarcoma on the distal limb of a warmblood foal using intralesional cisplatin chemotherapy.     

广灵驴分子系谱的建立及群体遗传结构分析

旨在对广灵县优种驴场保种群体进行调查的基础上构建分子系谱,并对其种群的遗传结构进行分析。本研究采集保种群成年、体况良好的广灵驴（体重350~400 kg）颈静脉血10 mL（n=107）,其中公驴13份,母驴94份,抗凝处理后提取全血DNA。采用12个微卫星标记进行荧光PCR扩增后,用ABI3730测序仪进行分型。分型结果采用Cervus 2.0和Pedigraph 2.4软件构建分子系谱,同时采用STRUCTURE2.3和Fstat软件计算群体遗传参数,采用R语言的hclust函数绘制7头公驴及其后代的系统发生NJ树（邻接树）。结果,对107头种驴进行了分子系谱构建,找到了30头子代的父亲和7头子代的母亲,系谱可靠性>90%;微卫星标记的平均观测杂合度（H_Obs）和多态信息含量（PIC）分别为0.676 5和0.593 9,标记遗传多样性较高;NJ树对7个公驴家系进行了聚类;群体分化系数（F_ST）为0.184,群体平均近交系数（F_IT）为0.033,群体内近交系数（F_IS）为-0.238,且群体处于哈代-温伯格平衡状态,存在很弱的近交。本研究建立了广灵驴保种群可靠性较高的分子系谱并对其遗传结构进行了分析,证明该群体遗传多态性较高,群体近交系数较低,处于较好的保种状态,具有较大的品种资源开发潜力。     

Arthrodesis for Congenital Flexural Deformity of the Metacarpophalangeal and Metatarsophalangeal Joints

A llama, a miniature horse, and a miniature donkey with severe bilateral congenital flexural deformities of the metacarpophalangeal and metatarsophalangeal joints were treated successfully by arthrodesis with dynamic compression plating or external skeletal fixation. The flexor tendons were more taut than the suspensory ligaments and were transected. In the llama, the suspensory ligament was transected, and overcorrection caused ischemic necrosis of one distal limb and subluxation of the other fetlock joint. In the horse and donkey, the suspensory ligament was preserved and wedge osteotomy was performed to correct the deformity.     

Congenital limb abnormality in a Thoroughbred foal in Trinidad

A congenital defect of the right hindlimb of a 36‐h‐old Thoroughbred foal as reported for the first time in Trinidad, West Indies. The foal was subjected to euthanasia and radiographs of both hindlimbs revealed that the bones distal to P1 of the right hindlimb only, were missing. Insertion of both the superficial and deep digital flexor tendons were found to be on the most proximal aspect of P1. Necropsy revealed no other congential defects.