Validation of the BioPRYN enzyme‐linked immunosorbent assay for detection of pregnancy‐specific protein‐B (PSPB) and diagnosis of pregnancy in American bison (Bison bison)

This study assessed the accuracy of the commercial BioPRYN^® ELISA for the detection of pregnancy‐specific protein‐B (PSPB) using a single blood sample to determine pregnancy status in American bison (Bison bison). A total of 49 bison cows were used in the study, and sampled at two time‐points during the gestation period, fall and spring, correlating with early‐ to mid‐term gestation (average 62.9 days post‐mating) and mid‐ to late‐term gestation (average 229.2 days post‐mating), respectively. Sensitivity of the test during early‐ to mid‐term gestation sampling period (fall) was 87.1%, while specificity was 100%; sensitivity of the test during late‐term gestation sampling period (spring) was 96.3%, while specificity remained at 100%. In total, the test showed a total sensitivity of 91.4%, specificity of 100% and total accuracy of 93.8%, similar to domestic cattle. Use of the single‐sample BioPRYN^® ELISA in American Bison for pregnancy diagnosis is economical and practical, minimizing animal handling time, frequency and subsequent stress while providing accurate results for pregnancy diagnosis at 62 days post‐mating. This method should be considered over more traditional pregnancy diagnosis methods for use in managed bison herds.     

Effect of the Medium Replacement Interval on the Viability,Growth and In Vitro Maturation of Isolated Caprine and Ovine Pre‐Antral Follicles

The aim of the present study was to evaluate the effects of different medium replacement intervals on the viability, antral cavity formation, growth and in vitro maturation (IVM) of oocytes from caprine and ovine pre‐antral follicles. Pre‐antral ovarian follicles (≥150 μm) were isolated from the ovarian cortex of goats and sheep and were individually cultured for 24 days using two different medium replacement intervals [2 days (T₁) or 6 days (T₂)]. Follicle development was evaluated on the basis of antral cavity formation, increases in follicular diameter and the presence of healthy cumulus oocyte complexes and fully grown oocytes. For caprine species, results showed a higher percentage (p < 0.05) of viable follicles in T₁ than T₂ from day 6 until the end of the culture. In addition, when comparing both treatments after the same culture duration, the rate of antrum formation was significantly higher in T₁ than in T₂ from day 12 onwards. Yet, in ovines, when both treatments were compared on day 24 of the culture, there were more viable follicles in T₂ than in T₁ (p < 0.05). In the caprine species, percentages of fully grown oocytes (≥110 μm) acceptable for IVM after 24 days of culture were significantly higher in normal follicles cultured in T₁ (30.0%) than in T₂ (6.7%; p < 0.05). On the other hand, in ovines, at the end of the culture, the percentage of oocytes destined for IVM was higher in T₂ than in T₁ (23.5% vs 2.9%; p < 0.05). In conclusion, under the same conditions, the frequency of medium replacement significantly affected the in vitro development of caprine and ovine pre‐antral follicles. To improve the efficiency of the culture system, the medium must be replaced every 2 and 6 days for goat and sheep pre‐antral follicles, respectively.     

Prevalence of anthelmintic resistance on sheep farms in New Zealand

AIM: To establish the prevalence of anthelmintic resistance in parasitic nematodes on sheep farms in New Zealand.

METHODS: A cross-sectional prevalence study was conducted, using a standardised faecal nematode egg count (FEC) reduction (FECR) test (FECRT) for ivermectin, at a full (0.2 mg/kg) and half (0.1 mg/kg) dose rate, and albendazole, levamisole and albendazole-levamisole in combination, on 60 lambs (n=10 per group) on farms selected from throughout New Zealand. Farms that conformed with selection criteria were chosen at random (n=80) or with a history of suspected resistance to macrocy- clic lactone (ML) anthelmintics (n=32). Resistance to an an- thelmintic was inferred when there was <95% reduction in FEC 7-10 days after treatment. Larval cultures were performed for all control groups and for treated groups for which resistance was evident.

RESULTS: Of the farms randomly selected, 36% showed ≥95% FECR for all anthelmintics tested; resistance to ivermectin at 0.1 and 0.2 mg/kg liveweight was evident on 36% and 25% of these farms, respectively. Resistance to both ivermectin (0.2 mg/kg) and levamisole was evident on 8/80 (10%) farms, to ivermectin and albendazole on 10/80 (13%) farms, and to iver- mectin, levamisole and albendazole on 6/80 (8%) farms. The prevalence of resistance to a half dose of ivermectin tended to be more prevalent on farms with a history of suspected ML resistance (p=0.06). Resistance to albendazole was seen across all the main parasite genera, and to levamisole in Nematodirus, Ostertagia (= Teladorsagia) and Trichostrongylus species. Resistance to ivermectin was dominated by Ostertagia spp, although Cooperia, Nematodirus and Trichostrongylus species were also implicated.

CONCLUSION: Anthelmintic resistance in parasitic nema-todes of sheep is common in New Zealand. Not only was resistance to albendazole and levamisole common, but resistance to the ML, ivermectin, was at a higher prevalence than expected. Sheep farmers and advisors in New Zealand need to re-evaluate the way they manage parasites, and more research is urgently needed if the steady decline in anthelmintic susceptibility is to be halted.     

MAGNETIC RESONANCE IMAGING FOR THE DIAGNOSIS OF STRESS FRACTURES IN A HORSE

In humans, magnetic resonance (MR) imaging is the method of choice for the diagnosis of stress fractures. In this paper, bilateral stress fracture of the lateral condyle of the third metacarpal bone in a French trotter is described. Results of the radiographic, MR imaging, and histologic examinations are presented, with a focus on the MR signal abnormalities found. Based on this patient, the potential use of MR imaging for the diagnosis of stress fractures in horses is discussed.     

In Vitro Comparison of a Novel External Fixator and Traditional Full‐Limb Transfixation Pin Cast in Horses

Objective: To compare the mechanical properties and failure modes of a standardized short oblique distal radial metaphyseal osteotomy stabilized using either a transfixation pin cast (TPC), a modular‐sidebar external skeletal fixator (ESF), or a solid‐sidebar ESF (modular‐ or solid‐ESF, respectively) using static or cyclic axial loading to failure. Study Design: In vitro study. Animals: Equine cadaver forelimbs. Methods: A 30° oblique distal radial osteotomy was created and stabilized using 1 of the 3 fixation methods: (1) TPC, (2) modular‐ESF, or (3) solid‐ESF. Limbs were tested using static (TPC, modular‐ESF, and solid‐ESF) or cyclic (TPC and solid‐ESF) axial loading to failure. The stiffness, yield load, yield displacement, failure load, and failure displacement for static loading and the cycles to failure for cyclic loading at 75% failure load were obtained. Data were analyzed using a Kruskal–Wallis test. Level of significance was P<.05. Results: The solid‐ESF had a greater stiffness, higher yield and failure load and a lower yield and failure displacement than the TPC (P=.01) and the modular‐ESF (P=.02). TPC had a higher yield load, failure load, and yield displacement than the modular‐ESF (P=.01). Mean cycles to failure for TPC was 2996±657 at a load of 16,000 N and for solid‐ESF 6560±90 cycles at a load of 25,000 N. Conclusions: The solid‐ESF was stiffer and stronger than the TPC and modular‐ESF and failed at a greater number of cycles in axial loading compared with the TPC. Clinical Relevance: This study is an initial step in evaluating the solid‐ESF. Further testing needs to be performed, but this fixation may offer a viable alternative to the traditional TPC for stabilization of long bone fractures in adult horses.     

Changes in Testicular Development, Ultrasonographic and Histological Appearance of the Testis in Buck Kids Immunized Against LHRH Using Recombinant LHRH Fusion Protein

This study was designed to evaluate the effectiveness of recombinant Ovalbumin-LHRL (OL) immunization on changes in testicular size, histological appearance and testosterone production in buck kids. Thirty native buck kids at 18 weeks of age were divided into three groups, control (n = 10), immunization (n = 10) and castration (n = 10) groups. Immunized animals received OL protein generated by recombinant DNA technology. Ultrasonographic and histological examinations of the testes were performed. Animals were slaughtered at 44 weeks of age. Semen and epididymides were evaluated for the presence of sperm cells. Immunized animals generated anti-LHRH antibodies. Testosterone production, testicular and accessory glands development and sperm production were suppressed in the immunized animals (p < 0.01). Semineferous tubule diameters decreased (p < 0.01), basal membrane of the tubule was thickened and hyalinized in immunized kids. Immunization affected ultrasonographic appearance of the testes drastically. While testes of control animals gained their normal ultrasonographic appearance as the age increased, immunized animals had uniform hypoechogenic testicular structure as observed at 18 weeks of age until slaughter. Simultaneous histological and ultrasonographic evaluations indicated that the changes in testicular histology could partly be monitored via ultrasonographic imaging; nevertheless, it is difficult to claim that ultrasonographic image reflects the exact changes in such instances. In conclusion, these results indicate that recombinant OL fusion protein is effective in immunocastration in buck kids and has a potential to be used as an alternative to physical castration. Further researches should be conducted to help assessing reproductive status of testes from ultrasound images.     

Third‐compartment cannulation in alpacas using a polyurethane gastrostomy tube

Objective To develop a simple and effective surgical technique for third‐compartment cannulation in alpacas. Design Prospective study using six adult male alpacas. Methods General anaesthesia was induced and a polyurethane gastrostomy tube was surgically implanted into the distal portion of the third compartment. Results Three of the alpacas retained their cannulas for a 100‐day period; however, three cannulas were dislodged during the study. Two of the three dislodged cannulas were replaced during a second surgical procedure. Cannulas were well tolerated by the alpacas and all animals remained clinically healthy during the study period. Third compartment contents did not leak from the cannulation site. The tubes were manually removed following the completion of the study and the small defect in the body wall quickly healed over in all animals. Conclusion Surgical placement of polyurethane tubes designed for percutaneous endoscopic gastrostomy is a useful method of cannulating the third compartment in camelids. This technique can be used for experimental studies and possibly could be used for nutritional support and fluid therapy in sick camelids that might need long‐term care.     

Congenital dyserythropoietic anaemia and dyskeratosis in Australian Poll Hereford calves

Congenital dyserythropoietic anaemia (CDA) is a heterogeneous group of rare genetic disorders that in humans is characterised by ineffective haematopoiesis with morphological abnormalities in erythroid precursor cells and secondary iron overload. In the 1990s, a syndrome of CDA with dyskeratosis and progressive alopecia was reported in Poll Hereford calves in Canada and the USA. We report the clinical and pathological findings in two Poll Hereford calves with this syndrome from separate properties in South Australia. The animals had a variably severe anaemia, associated with abnormal nucleated red blood cells in peripheral blood, and large numbers of rubricytes and metarubricytes with a characteristic nuclear ultrastructure in the bone marrow. Both calves were born with a wiry hair coat and a progressively ‘dirty‐faced’ appearance associated with hyperkeratosis and dyskeratosis (apoptosis).     

A comparison of ultra-high-molecular weight polyethylene cable and stainless steel wire using two fixation techniques for repair of equine midbody sesamoid fractures: an in vitro biomechanical study

OBJECTIVE: To compare the monotonic tensile and fatigue strength of 16-gauge stainless steel wire (SSW) to ultra-high-molecular weight polyethylene (UHMWPE) cable using a transfixed cerclage technique in an in vitro midbody sesamoid osteotomy model. Endoscopic modifications to Martins transfixed cerclage technique were developed. A new suture technique of fixation was compared with the transfixed cerclage technique by measuring gap formation after cyclic testing. STUDY DESIGN: An in vitro biomechanical paired equine cadaver limb study. SAMPLE POPULATION: Twenty-one paired cadaveric adult equine forelimbs. METHODS: Uniaxial medial midbody sesamoid osteotomies were created in paired adult equine forelimbs. Monotonic tensile strength was measured on 10 forelimbs repaired by a transfixed cerclage technique using wire or cable. Fatigue testing to failure was performed on 4 forelimbs repaired using the transfixed cerclage technique by cycling the limbs between 500 N and 2,000 N. The limbs were initially repaired with wire, cycled until the wire broke, then repaired with cable and cycled again to failure. Fatigue testing for gap displacement was performed on 8 limbs repaired with either the transfixed cerclage technique or the suture technique. Limbs were cycled between 500 N and 2,000 N for 10,000 cycles. The limbs were repaired with wire initially, tested, and then repaired with cable and tested again. Twenty-two limbs were used for mechanical testing. The remaining limbs (20) were used to develop and practice the endoscopic transfixed cerclage (10 limbs) and suture (10 limbs) techniques. RESULTS: Ultimate tensile strength (UTS) of UHMWPE cable constructs was 34% greater than the UTS of SSW constructs. Fatigue strength was 2 to 20 times greater for UHMWPE cable constructs than SSW constructs. Separation of fragments was 153% less for limbs repaired by the suture technique compared with those repaired by the transfixed cerclage technique. CONCLUSIONS: UHMWPE cable shows promise for this clinical application because of its greater tensile and fatigue strength. The newly described suture technique significantly reduced gap formation compared with the transfixed cerclage technique. Osteotomy gap formation occurred early in cycling, suggesting that rigid support in the form of a cast may be needed during the early postoperative period for wiring techniques. CLINICAL RELEVANCE: Clinical testing of UHMWPE cable should eliminate problems of wire breakage seen with SSW. The endoscopic transfixed cerclage technique can be used by surgeons familiar with arthroscopic surgery. However, the suture technique needs to be tested in vivo to determine whether there is a clinical advantage compared with the transfixed cerclage technique.