Heterotopic Transfer of Fresh and Cryopreserved Autogenous Articular Cartilage in the Horse

Two 10 mm thick osteochondral grafts were harvested from the lateral aspect of the lateral trochlear ridge of the left talus in each of 10 anesthetized horses. The grafts were frozen in a 7.5% DMSO solution and stored in liquid nitrogen. The horses were anesthetized again on day 14 and the thawed grafts were press-fitted into drill holes in the trochlear ridges of the right stifle. A fresh graft was transferred from the right hock to the left stifle. To control for the effects of surgery, another fresh graft was transferred from the right stifle to the left stifle. The result was two grafts in each femoropatellar joint. Fresh and frozen osteoarticular autografts appeared to maintain a durable weight-bearing surface for 3 months; however, the fresh grafts were clearly superior. Frozen grafts had fewer living chondrocytes, decreased safranin-O staining, and decreased SO435 uptake. Graft stability and articular surface congruency were determining factors in the outcome of all grafts. Since the availability of osteochondral autografts is limited, further work on the use of preserved allogeneic osteochondral tissue is warranted.     

Cortical Bone Allografting in the Horse

Half and full cylindrical frozen cortical bone allografts were transplanted into groups of horses. All horses accepted, resorbed, and began replacement of their grafts to varying degrees, according to frequent radiographic and monthly histologic evaluation. Clinically, all the recipients appeared to return to varying degrees of function. This method of bone allografting is suggested as an adjunct to current methods of fracture fixation in the horse.     

Multiple Osteochondral Autografts for Treatment of a Medial Trochlear Ridge Subchondral Cystic Lesion in the Equine Tarsus

Objective— To adapt the multiple osteochondral autograft technique for treatment of a subchondral cystic lesion (SCL) of the proximal medial trochlear ridge (MTR) of the equine talus and assess long term outcome.
Study Design— Case report.
Animals— Quarter horse stallion with SCL of the proximal MTR of the talus.
Methods— Osteochondral autograft techniques used in dogs and humans were adapted and optimized in equine cadavers. A horse with a SCL of the MTR of the talus was treated by osteochondral autografting. Three osteochondral autografts were harvested from the distal lateral trochlear ridge of the talus in the affected tarsus. No curettage or debridement of the osteochondral lesion was performed. Three recipient beds were predrilled and osteochondral autografts were press-fit into the proximal MTR of the talus. Outcome was assessed by radiography, telephone interview of the owner, and direct observation during training.
Results— Postoperative radiographs indicated excellent filling of the osteochondral defect and graft-articular surface congruency. No complications were encountered after surgery. At 10 months after surgery, the horse successfully re-entered reining training, and continued to be sound and performing athletically 2 years after surgery.
Conclusion— Osteochondral autografting deserves consideration and further evaluation as a primary treatment option for SCL of the proximal MTR of the talus in horses.
Clinical Relevance— Based on the outcome of this case, osteochondral autografting appears to be a safe, valuable and realistic option for treating SCL of the proximal MTR of the talus in equine athletes.     

Evidence of Damage in Cryopreserved and Fresh Bovine Embryos Using the Tunel Technique

The objective of the present study was to evaluate the quality of bovine embryos cryopreserved in different years in Chiapas, Mexico. The embryos were obtained from a government institution (FIMEGEN) dedicated to promoting embryo transfer among dual-purpose cattle farmers. Forty-three embryos frozen in 1988, 1989, 2000 and 2002 were analysed with the Tunel technique to detect programmed cell death (apoptosis). Eleven fresh embryos were used as controls. Analysis of variance was used in embryos stored in the different years with averages tested using Tukey's test. Student's t-test was employed to compare fresh and frozen cells. Embryos with shorter storage time presented a lower number (p < 0.001) of Tunel-positive cells compared with embryos stored for longer time. On the contrary, when comparing the number of apoptotic cells between frozen and fresh embryos a higher number of positive cells (p < 0.05) were found in the former. The present results suggest that the cryopreservation per se caused damage that compromises the viability of the embryo. Another explanation for the lower pregnancy rate found in the tropics could be irreversible damage caused by poor storage technique in these large operations.     

Arthroscopic Removal of an Osteochondral Fragment from the Middle Phalanx of a Horse

An intraarticular osteochondral chip fracture of the distal dorsolateral aspect of the right hind middle phalanx in a 4-year-old Dutch Warmblood gelding was removed arthroscopically. Accessibility and visibility of the fragment were excellent, and there was minimal soft tissue trauma.     

Effect of Seminal Plasma Components on the Quality of Fresh and Cryopreserved Stallion Semen

The importance of seminal plasma (SP) components for stallion semen quality and freezability is little known. This study aimed to evaluate the relationship between SP components and fresh/cryopreserved stallion semen quality. Semen of 30 stallions was collected, and then, SP was recovered and lyophilized. Total protein (TP), vitamin C (CVIT), vitamin E (EVIT), vitamin A (AVIT), iron (Fe), copper (Cu), magnesium, and zinc (Zn) in SP were assessed. Sperm was frozen in an extender supplemented with lyophilized SP. In fresh semen motility, abnormal morphology (AM), sperm vitality (SV), and plasma membrane integrity (PMI) were evaluated. In post-thaw semen, additionally, total motility (TM), progressive motility (PM), straight line velocity (VSL), curvilinear velocity (VCL), average path velocity (VAP), amplitude of lateral head displacement (ALH), and beat cross-frequency (BCF) were assessed. Levels of component of SP were established by a distribution analysis. Generalized linear models were fitted. Comparisons of means were done with Tukey's test. Correlation and regression analyses were performed. Vitamins and ions were found to be related to fresh semen quality. For post-thaw sperm, medium TP showed higher semen quality. Negative regression and correlation coefficients between CVIT and all post-thaw semen parameters were found. Low EVIT yielded the lowest PM, VSL, and VAP values, while a high level of AVIT yielded the best results for sperm quality. A high level of Cu yielded higher results for TM, PM, VCL, and ALH. Moreover, a negative correlation was found between Zn, SV, and PMI. In conclusion, SP composition influences fresh and post-thaw stallion semen quality.     

Radial-intermediate Carpal Bone Fusion in a Horse

The radial carpal and intermediate carpal bones were found to be fused in a four year old Quarterhorse stallion having a subtle forelimb lameness. Clinical examination, diagnostic local anesthesia, radiography and arthroscopy were used to arrive at the diagnosis. A cause could not be established, although perinatal deformation of cartilagenous carpal bone templates is suspected. A similar case of carpal bone fusion was found among carpi collected for an anatomical study, indicating this condition may be more common than expected. Clinicians should carefully evaluate the flexed lateral view of the carpus for proximal movement of the intermediate and ulnar carpal bones in young horses presented for carpal lameness. The significance of this condition will remain uncertain until its incidence and relationship to lameness are better defined.     

Surgical Management of Proximal Splint Bone Fractures in the Horse

Fractures of Metacarpal and Metatarsal II and IV (the splint bones) were treated in 283 horses over an 11 year period. In 21 cases the proximal portion of the fractured bone was stabilized with metallic implants. One or more cortical bone screws were used in 11 horses, and bone plates were applied in 11 horses. One horse received both treatments. Complications of screw fixation included bone failure, implant failure, radiographic lucency around the screws, and proliferative new bone at the ostectomy site. Only two of the horses treated with screw fixation returned to their intended use. Complications of plate fixation included partial fixation failure (backing out of screws), wound drainage, and proliferative bony response around the plate. Six of the 11 horses treated by plate fixation returned to their intended use. The authors recommend consideration of plate fixation techniques for repair of fractures in the proximal third of the splint bone.     

不同糖类稀释液对低温保存杜泊羊鲜精的影响

为了比较添加不同糖类(葡萄糖、果糖、D-半乳糖和蔗糖)稀释液对杜泊羊鲜精品质的影响,试验选用3岁健康杜泊种公羊3只,采集精液,等量分装,分别加入等量等温的含不同糖类的精液稀释保存液,并在0h、6h、24h、30h、48h、54h、72h和78h对精子活率、质膜完整率和顶体完整率进行检测。结果表明,混合糖精子活力、质膜完整率和顶体完整率都最高,与其他组差异显著(P0.05);D-半乳糖组质膜完整率比葡萄糖组和果糖组高,差异显著(P0.05),其他指标三者差异不显著(P0.05);蔗糖组效果较差,与其他组差异显著(P0.05),对照效果最差,与其他组差异极显著(P0.01);有效存活时间和生存指数混合糖组最高。结果提示:添加葡萄糖、果糖和D-半乳糖的混合糖作为杜泊羊新鲜精液的保存液效果较好。     

Osteochondral Fragments within the Dorsal Pouch or Dorsal Joint Capsule of the Proximal Intertarsal Joint of the Horse

The anatomy of the dorsal pouch of the proximal intertarsal joint (PIJ) and its communication with the tarsocrural joint (TCJ) was studied in 15 pairs of hocks from young and mature horses. The mediolateral length of the TCJ-PIJ fenestration was 14 to 29 mm. The potential volume of the dorsal pouch of the PIJ was 3 to 5 ml, and a recess extended 10 to 28 mm medial to the medial commissure of the TCJ-PIJ fenestration. In a correlated clinical study, osteochondral fragments were identified radiographically within the dorsal pouch (category 1) or dorsal joint capsule (category 2) of the PIJ in 17 horses undergoing arthroscopic surgery of the TCJ. In six horses with category 1 lesions, osteochondral fragments were found free within the dorsal pouch and were removed. In five horses, category 1 fragments were not located. All 11 horses were reported by owners to be sound after surgery. In seven horses, a minimally displaced fragment was identified at the distal aspect of the medial trochlear ridge, within the insertion of the synovium separating TCJ and PIJ (one horse had both types of lesions in the same hock). Five of the seven category 2 fragments were removed at surgery. Of the seven horses with category 2 lesions, four were training or racing, two were unsound, and one was still convalescing at the time of follow-up.     

Pregnancy in the Caspian Miniature Horse Using Frozen Semen Cryopreserved with the EquiPRO CryoGuard Freeze Medium and Customized Freezing Protocols

The primary goal of this project was to establish a protocol to freeze the sperm of the Caspian miniature horse in an attempt to start an intensive artificial insemination program to effectively increase the population of this breed, which has been listed as “Critical Rare Breed” by the American Livestock Breed Conservancy and is in danger of extinction. Commercially available equine freezing medium (EquiPRO CyoGuard Complete egg-yolk extender) was used for the initial setup of two different freeze protocols: slow and fast. The fast-freeze protocol had slightly better postthaw results and was used for a fertility demonstration. Five mares of proven fertility, aged 3 to 12 years, were used in the fertility trials, two of which resulted in pregnancy. This is the first report of pregnancy in the Caspian miniature horse using frozen semen, and the results seem to be a promising start to an extensive program to help this endangered breed, although further research on freezing protocols and conditions for this process are necessary to further improve the survival of semen and pregnancy rate.     

A Large Frontonasal Bone Flap for Sinus Surgery in the Horse

A large frontonasal bone flap was created to treat diseases of the paranasal sinuses in 14 horses. The bone flap was made as wide as possible within the confines of the nasolacrimal duct so the floor of the frontal sinus and the dorsal and ventral conchae could be opened. These openings exposed the nasal passages, maxillary sinuses, and ventral conchal sinus thereby facilitating removal of diffuse and localized lesions from these sites. Diseases treated were ethmoid hematomas (4 horses), sinus cysts (5 horses), cryptococcal granuloma, osteoma, hemangiosarcoma, pus in the ventral conchal sinus, and periapical infection of a second molar. Four horses were euthanatized during or after surgery, one because of postsurgical pleuritis and pneumonia (horse with osteoma) and three because of their primary problems (cryptococcal granuloma, hemangiosarcoma, pus in the ventral conchal sinus). Skin suture abscesses that responded to treatment developed in four horses. Ten horses returned to their intended uses, the sinus flaps healed without blemish, and the original problems did not recur. The frontonasal flap technique provided greater access to all paranasal sinuses than methods described previously.     

门源马骨营养状况的检测

对门源县 3 8匹马的骨营养状况指标进行了检测。结果发现 :门源马的血清钙浓度为 2 .68mmol L ,血清无机磷浓度为 1.3 6mmol L ,血清碱性磷酸酶活性为 66.0 4IU L ,血清羟脯氨酸含量为 15 .2 6μmol L ,均处于正常的范围。按性别组间比较 ,公马的血清钙浓度显著高于母马的 (P <0 .0 5 ) ,其他各项指标均无显著性差异 (P >0 .0 5 )。     

Evaluation of the Tibia as a Source of Autogenous Cancellous Bone in the Horse

A 1 cm defect was created in the proximal medial cortex of 12 tibiae, cancellous bone was removed from the site, and the bones were fractured by loading in torsion. The fractures did not occur through the donor sites and the fracture patterns and loads to fracture were similar for the treated tibiae and their untreated mates. Cancellous bone was harvested from the proximal medial aspect of both tibiae in nine adult horses. The soft tissue wounds were monitored for more than 10 days in seven horses and healing of the osseous defects was evaluated radiographically at regular intervals to month 6 in two horses. The site provided convenient access with the horses in lateral or dorsal recumbency, the anatomic landmarks were reliable, and the surgical procedure was simple and short. Complications occurred in only three incisions.     

Lag Screw Fixation of a Sagittal Fracture of the Third Carpal Bone in a Horse

A sagittal fracture of the third carpal bone of a Standardbred racehorse was repaired with lag screw fixation. Healing of the fracture was uncomplicated, and the horse made a successful return to racing. It was concluded that selected sagittal fractures of the third carpal bone may be treated successfully with lag screw fixation. Criteria for selection of cases include fragment size and orientation of the fracture plane.     

Osteochondral Defects of the Lateral Trochlear Ridge of the Distal Femur of the Horse Clinical, Radiographic, and Pathological Examination and Results of Surgical Treatment

Osteochondral defects of the lateral trochlear ridge of the distal femur were identified in 15 joints of 10 horses. Lesions were bilateral in five horses and unilateral in five horses. Thirteen of these 15 defects were treated by curettage through a craniolateral femoropatellar arthrotomy. Subcutaneous seroma formation and partial skin dehiscence occurred in nine stifles. Lameness attributable to the stifle joint was no longer apparent at 6 and 12 months after surgery. The normal subchondral bony contour of the lateral trochlear ridge was altered in all joints after surgery. The subchondral bone was uniformly increased in density in all joints, and six lateral trochlear ridges had small focal radiolucent regions within the subchondral bone, which suggested that complete removal of the original lesion had not been achieved. Healing of the surgical site between 2 weeks and 14 months was evaluated in four joints. Granulation tissue from mesenchymal elements in subchondral marrow spaces gradually filled the defects with fibrocartilage.     

Production of Lambs After the Transfer of Fresh and Cryopreserved in vitro Produced Embryos from Prepubertal Lamb Oocytes and Unsorted and Sex-sorted Frozen-thawed Spermatozoa

Cumulus-oocyte complexes from hormone-stimulated 3-4-week-old (n=43) and 6-7-week-old (n=12) prepubertal lambs were matured in vitro and incubated with unsorted, or X- or Y-spermatozoa separated with a high-speed cell sorter (SX MoFlo)frozen-thawed. Presumptive zygotes were then cultured to the blastocyst stage, and transferred to recipients fresh or after cryopreservation (frozen). Oocyte cleavage was higher (p <0.05) with unsorted (515/926, 55.6%) than X- or Y-spermatozoa (261/672, 38.8% and 229/651, 35.2%, respectively) and blastocyst formation (% zygotes) by Day 9 of in vitro culture was lower (p <0.05) for X- (102/261, 39.1%) than unsorted spermatozoa (249/515, 48.3%), but did not differ between Y-spermatozoa (103/229, 45.0%) and unsorted spermatozoa, or between X- and Y-spermatozoa (p >0.05). For fresh embryos, survival to term was 50.0% (3/6) for unsorted, 0.0% (0/6) for X- and 16.7% (1/6) for Y-spermatozoa-derived embryos (p >0.05), and for frozen embryos was 4.0% (2/50) for unsorted, 9.1% (2/22) for X- and 2.9% (1/34) Y-spermatozoa-derived embryos (p >0.05). Of the two lambs born from X-spermatozoa-derived embryos, one was female (50%), and from the two Y-spermatozoa-derived lambs, both were male (100%), demonstrating that lambs can be produced after the transfer of fresh and cryopreserved IVP embryos derived from prepubertal lamb oocytes and frozen-thawed sex-sorted sperm.     

Dorsal Laminectomy in the Horse II. Evaluation in the Normal Horse

Subtotal dorsal cervical laminectomy was performed on nine normal horses to evaluate positioning, refine the surgical technique, and assess the immediate and long-term effects of this procedure in the horse. Funkquist type B laminectomies were performed at various levels from C3-C4 to C6-C7. Absorbable gelatin sponge or autogenous nuchal fat was placed over the exposed dura mater. Surgery times varied from 2 to 3 hours, and all horses recovered without complication. Rapid primary intention wound healing occurred, with two horses showing neck pain. All remained neurologically normal. The horses were euthanized 6 weeks, 3 months, or 6 months postoperatively. Myelograms performed just prior to euthanasia demonstrated normal or expanded dye column widths. Necropsy revealed that all surgical sites had healed well. The laminectomy membrane was soft and pliable and could be easily separated from the underlying dura mater when fat was used as interpositional material. When gelatin sponge was used, the laminectomy membrane was firmly bonded to the dura. Spinal cords at the level of laminectomy appeared normal and moved freely within the vertebral canal. Histological evaluation of the laminectomy sites and associated spinal cord revealed no abnormalities attributable to the surgery. It was concluded that dorsal cervical laminectomy can be performed in horses without untoward sequelae. The procedure may be useful for the treatment of equine cervical compressive myelopathies.