Application of Platelet-Rich Gel to Enhance Wound Healing in the Horse: A Case Report

A large torn wound of the dorsal elbow region was observed in a 17-year-old Arabian mare. Surgical reconstruction was performed with the horse in standing sedation, but suture dehiscence occurred 2 days later. Autologous platelet-rich gel (PRG) was then applied to the wound every 3 weeks for a total of 3 administrations to accelerate good-quality healing. The wound had healed rapidly and completely within 5 months of the first PRG treatment, without chronic effects or formation of exuberant tissue granulation and with minimum scarring. This case report suggests that topical treatment with autologous PRG, as additional therapy, might be considered beneficial in the management of large-wound healing in horses, and it can be regarded as safe and inexpensive treatment that can be used in field.     

A Novel Bioelectric Device Enhances Wound Healing: An Equine Case Series

The use of low-level microcurrents for accelerated wound healing is well documented. A case series was conducted to assess wound healing outcomes following the application of a wireless, current-generating bioelectric wound care device in 10 equines that presented with traumatic injuries in the lower extremity. Wounds were treated with a bioelectric device held in place with standard bandaging twice a week. At each follow-up visit, wounds were photographed and assessed for signs of epithelialization. All presented wounds were reduced in size or achieved complete wound closure, with an average 1.3% wound healing per day. The results of this case series demonstrate the safety and efficacy of a bioelectric device as a management option for traumatic lower-extremity wounds in equines and hold significant promise in promoting enhanced healing rates and improved aesthetic outcomes.     

Campylobacter Cross‐Contamination of Chicken Products at an Abattoir

Consumption of raw or undercooked poultry products contaminated with Campylobacter has been identified as a risk factor for human campylobacteriosis. We determined whether slaughtering of Campylobacter‐positive flocks was associated with contamination of chicken products derived from Campylobacter‐negative flocks slaughtered at the same abattoir. The presence of Campylobacter was investigated in 22 broiler farms 1 week prior to slaughter and in one abattoir on nine separate slaughter days. A total of 600 bulk packed chicken products were tested, with 198 (33.0%) of the products found to be Campylobacter positive. Of the 350 chicken products originating from Campylobacter‐positive flocks, 180 (51.1%) were contaminated with the bacteria. In contrast, only 18 (7.2%) of 250 chicken products derived from Campylobacter‐negative flocks were contaminated. In 14 of these 18 products, the Campylobacter isolates were identical to isolates obtained from the flock slaughtered immediately prior to the Campylobacter‐negative flock. Notably, on 4/6 slaughter days, Campylobacter‐negative flocks were slaughtered prior to the positive flocks, and Campylobacter was absent from all chicken products originating from the negative flocks. These results suggest that implementation of logistic slaughter (where Campylobacter‐negative flocks are slaughter first) significantly decreases the prevalence of Campylobacter‐positive chicken products.     

鸡蛋提取物角蛋白及鸡蛋黄对兔伤口愈合的临床研究

为了探讨鸡蛋提取物角蛋白及鸡蛋黄对伤口愈合影响的临床研究,取健康家兔12只,随机分为A、B两组,每组6只,在每只家兔脊背两侧设试验伤口和对照伤口各1个,即同体不同部位对照试验;A组试验处伤口涂抹鸡蛋提取物角蛋白;B组试验处伤口涂抹生鸡蛋黄;A、B两组对照处伤口均未作任何处理。结果表明:A组和B组涂抹鸡蛋提取物角蛋白和生鸡蛋黄的伤口愈合时间比未做任何处理的伤口愈合快3～4 d,无论是愈合速度还是愈合效果均非常明显且伤口感染率减少,伤口愈合的疤痕减淡,外部更加美观自然,且能达到祛疤美容效果。     

Immunoglobulin G Class Identification from Wild Ungulates by Cross‐reactivity with Antisera to Domestic Animals

Seven species of Spanish ungulates were tested for the presence of homologous immunoglobulin G (IgG) with a gel‐diffusion test using bovine, ovine, caprine and porcine IgG antisera. Homologous ovine and caprine IgG were detected in sera from chamois (Rupicapra pyrenaica), Spanish ibex (Capra pyrenaica hispanica), mouflon (Ovis orientalis musimon), red deer (Cervus elaphus), fallow deer (Dama dama) and roe deer (Capreolus capreolus). Homologous porcine IgG was detected in wild boar (Sus scrofa) serum. Immunoelectrophoretic assays were performed to compare the electrophoretic mobility of IgG from domestic and wild species.     

Effects of Nylon and Polyglactin 910 Suture Material on Perilimbal Corneal Wound Healing in the Dog

The effects of fine suture materials on corneal wound healing in the dog were studied. In 20 dogs, standardized perforating perilimbal clear corneal wounds were made and closed with either monofilament Polyglactin 910 or nylon. Five dogs each were euthanized 8,12,16, and 21 days postoperatively. Results of gross, biomicroscopic, and histologic examinations at 8,12,16, and 21 days showed nylon and Polyglactin 910 to cause similar inflammatory responses. Epithelialization and suppuration around the suture tracts were observed more frequently when Polyglactin 910 was used. Both materials were associated with a foreign body (granulomatous) response. There was no loss of wound integrity with either material. Strengths of the incised corneal tissues with and without the sutures intact were determined. At day 16 postsurgery, there was a statistically significant difference in tissue strengths between cases in which sutures were and were not intact. This difference was not apparent at 21 days postsurgery, which suggested that the suture dependent phase ends between the 16th and 21st postsurgical days. In addition, at day 16 there was no statistical difference between the tissue strength of wounds sutured with nylon or Polyglactin 910. Corneal suture materials studied should remain in place for at least 16 days, and absorbable material is only appropriate if it retains tensile strength for 16 days.     

Pathogenesis and Subsequent Cross‐Protection of Influenza Virus Infection in Pigs Sustained by an H1N2 Strain

The H1N1, H3N2 and, more recently, H1N2 subtypes of influenza A virus are presently co‐circulating in swine herds in several countries. The objectives of this study were to investigate the pathogenesis of Sw/Italy/1521/98 (H1N2) influenza virus, isolated from respiratory tissues of pigs from herds in Northern Italy, and to evaluate its potential cross‐protection against the Sw/Fin/2899/82 (H1N1) strain. In the pathogenesis test, eight pigs were intranasally infected with H1N2 virus; at pre‐determined intervals, these animals were killed and necropsied, along with eight uninfected animals. In the cross‐protection test, sixteen pigs were infected by intranasal (i.n.) and intratracheal (i.t.) routes with either H1N2 or H1N1 virus. Twenty days later, all pigs were challenged (by the same route), with either the homologous H1N2 or heterologous H1N1 virus strains. Control group was inoculated with culture medium alone. On post‐challenge days (PCD) 1 and 3, two pigs from each infected group, along with one control pig, were killed. Clinical, virological, serological and histopathological investigations were performed in both the pathogenicity and cross‐protection tests. In the pathogenicity test, mild clinical signs were observed in two pigs during 3 and 4 days, respectively. Virus was isolated from two pigs over 6 days and from lung samples of pigs killed on post‐infection days 2 and 4. Seroconversion was detected in the two infected animals killed 15 days after infection. In the cross‐protection study, mild clinical respiratory signs were detected in all pigs infected with either the H1N2 or H1N1 virus. The virus was isolated from nasal swabs of almost all pigs till 6 days. After the challenge infection, the pigs remained clinically healthy and virus isolation from the nasal secretions or lung samples was sporadic. Antibody titres in H1N1 or H1N2 infected groups were similar, whereas the H1N2 sub‐type induced less protection against re‐infection by homologous and heterologous virus than H1N1 sub‐type. The controls had no signs of the disease. In the H1N2 infected pigs, a reduced number of goblet cells in nasal and tracheal mucosa and small foci of lymphomononuclear cell infiltrates in the submucosa were detected. Furthermore, the goblet cell reduction was related to the time of infection. Diffuse mild interstitial pneumonia was also recorded in pigs infected with the H1N2 virus and challenged with either H1N1or H1N2 pigs. These studies showed the moderate virulence of the H1N2 virus and a partial cross‐protection against heterologous infection.     

Factors Associated with Dog Rabies Vaccination in Bhol,Philippines: Results of a Cross‐Sectional Cluster Survey Conducted Following the Island‐Wide Rabies Elimination Campaign

The Philippines has a long history of rabies control efforts in their dog populations; however, long‐term success of such programmes and the goal of rabies elimination have not yet been realized. The Bohol Rabies Prevention and Elimination Program was developed as an innovative approach to canine rabies control in 2007. The objective of this study was to assess canine rabies vaccination coverage in the owned‐dog population in Bohol and to describe factors associated with rabies vaccination 2 years after implementation of the programme. We utilized a cross‐sectional cluster survey based on the World Health Organization’s Expanded Programme on Immunization coverage survey technique. We sampled 460 households and collected data on 539 dogs residing within these households. Seventy‐seven per cent of surveyed households reported owning at least one dog. The human‐to‐dog ratio was approximately 4 : 1, and the mean number of dogs owned per household was 1.6. Based on this ratio, we calculated an owned‐dog population of almost 300 000. Overall, 71% of dogs were reported as having been vaccinated for rabies at some time in their lives; however, only 64% of dogs were reported as having been recently vaccinated. Dogs in our study were young (median age = 24 months). The odds of vaccination increased with increasing age. Dogs aged 12–23 months had 4.6 times the odds of vaccination compared to dogs aged 3–11 months (95% CI 1.8–12.0; P = 0.002). Confinement of the dog both day and night was also associated with increased odds of vaccination (OR = 2.1; 95% CI 0.9–4.9; P = 0.07), and this result approached statistical significance. While the programme is on track to meet its goal of 80% vaccination coverage, educational efforts should focus on the need to confine dogs and vaccinate young dogs.     

Isolation,Identification and Enrichment of Type A Spermatogonia from the Testis of Chinese Cross‐Bred Buffaloes (Swamp × River)

The proportion of type A spermatogonia in the isolated testis cells is a prerequisite for conducting experiments and the manipulation of these germ cells. Thus, this study was designed to examine the wide range of strategies for the isolation, identification and enrichment of type A spermatogonia in pre‐pubertal buffalo calves (3–6 months). Histological findings revealed the presence of maximum number of type A spermatogonia at 5 months, which was further confirmed by DBA immunohistochemistry. In a newly modified strategy for the isolation of testis tissues, mincing followed by trituration and two rounds of digestion with collagenase, hyaluronidase and DNase yielded more than 95% testis cell population. Differential plating with laminin, poly‐l ‐lysine and gelatin significantly (p < 0.05) affected the purity of type A spermatogonia. Among these extracellular matrix (ECMs) molecules, laminin and gelatin performed well and reached at a purity of 39.38 ± 1.21% and 32.15 ± 1.60%, respectively. In addition, combination of laminin and gelatin followed by Percoll centrifugation performed the best and yielded >90% type A spermatogonial purity. Moreover, viability of the cells was not affected (p > 0.05) irrespective of different enrichment methods. In conclusion, type A spermatogonia isolation and enrichment system was developed using different ECM molecules in buffaloes, which will aid in solving wide range of problems especially fertility‐related problems and transgenic animal production in buffaloes.     

The Effect of Trans-Stifle External Skeletal Fixation and Hyaluronic Acid Therapy on Articular Cartilage in the Dog

Transarticular external skeletal (TES) fixators were applied unilaterally to the stifle joints of 10 young adult dogs. After 4 weeks, the fixators were removed from all dogs. Two dogs were not allowed a remobilization period, whereas 8 dogs were provided with 4 additional weeks of weight-bearing activity in a kennel run. Four dogs were given high-molecular weight hyaluronic acid by intra-articular injection weekly during the remobilization period. Clinical gait evaluations and range of motion were determined during the remobilization period. Articular cartilage samples from both stifle joints of all dogs were evaluated histologically and histochemically. No significant differences in gait scores or range of motion were noted between treated and untreated dogs. Articular cartilage proteoglycan content was reduced after 4 weeks of trans-stifle external skeletal fixation as determined by loss of alcian blue (AB) histochemical staining. Improved homogeneity of histochemical staining was observed after remobilization. However, remobilization was associated with histological damage to the surface and tangential layers of articular cartilage. Remobilization combined with hyaluronic acid (HA) therapy improved histochemical staining and reduced structural damage to articular cartilage when compared with remobilization alone.     

透明质酸对猪精子活率,获能,顶体反应及体外受精穿透率的影响

采用具有高繁殖力的种公猪的新鲜精液及冷冻精液，加入不同获能剂咖啡因及透明质酸，在３９．５℃、５％ＣＯ２培养箱中孵化培养。结果表明，透明质酸能够在体外条件下保持精子的活率，并维持在稳定的水平，鲜精为４２．６％～４７．８％，冻精为２０．４％～２５．６％。荧光剂Ｈｏｅｃｈｓｔ染色表明，透明质酸能够显著增加培养３６０ｍｉｎ后的活精子率。在无蛋白来源的ｍＢＯ培养液中，培养０～６０ｍｉｎ，精子发生自发的获能和顶体反应，而受获能剂影响，精子发生获能的时间带在各处理组之间显著不同：咖啡因处理组的获能时间带是６０～３６０ｍｉｎ，透明质酸处理组则是６０～１８０ｍｉｎ。体外受精试验表明，去除卵母细胞周围卵丘细胞时，透明质酸和咖啡因对精子穿透率的影响没有区别；而在卵丘细胞存在的前提下，咖啡因处理组的穿透率显著高于透明质酸处理组。     

Alkaline and Acid Phosphatase, β‐Glucuronidase and Electrolyte Levels in Fractionated Stallion Ejaculates

Seminal plasma (SP) is a mixture of contents from the testes, epididymides and accessory sex glands. The sperm concentration is highest in the first few jets, or fractions, of the ejaculate, and the composition of SP varies between these fractions because accessory gland secretions are released in a specific order. The aim of this study was to compare the levels of Na, Cl, K, Mg, Ca, inorganic phosphate (Pi) and the enzymes alkaline phosphatase (AP), acid phosphatase (ACP) and β‐glucuronidase (BG) in the different fractions of the ejaculate and in different stallions. All semen collections were done using a computer‐controlled phantom that collects the ejaculatory jets separately in five cups. The cups with the highest (HIGH) and the lowest (LOW) sperm concentration were analysed. In Trial I, semen was collected from three reproductively normal stallions. In Trial II, ejaculates of two reproductively normal stallions were compared to those of two subfertile stallions. In Trial III, semen was collected from seven stallions with varying reproductive history. The sperm‐rich fractions contained the highest levels of AP, ACP, BG and inorganic phosphate, and the values were positively correlated to the sperm concentration. Significant differences between the subfertile and the fertile stallions pairs in HIGH : LOW ratios were found in Pi and Cl concentrations. The highest concentrations of Ca and Mg were found in the last fractions with low sperm concentrations, with no significant differences between the fertile and the subfertile stallion pairs. The concentrations of K, Na and Cl were similar in HIGH and LOW fractions and in whole ejaculate samples. Pre‐sperm fluid contained the highest concentrations of Na and Cl. Some of the possible variation in storage tolerance between ejaculates and ejaculatory fractions could perhaps be explained by differences in the composition of SP.     

Effects of N‐acetyl‐cysteine and N‐acetyl‐cysteine‐amide Supplementation on In Vitro Matured Porcine Oocytes

This study was conducted to evaluate the effects of different concentrations of the antioxidant N‐acetyl‐cysteine (NAC) supplemented to the maturation medium on porcine embryo development. Concentrations of NAC and its synthetic derivative, NAC‐amide (NACA) were evaluated for effects on nuclear maturation, fertilization success and embryo development. Concentrations of NAC (0, 0.5, 1.0, 1.5, 2.0, 2.5 and 5.0 mm ) were supplemented to maturing oocytes, and embryo development was analysed at 48 and 144 h post‐fertilization. There were no differences among cleavage rates for any of the treatment groups. Blastocyst formation for 1.5 mm NAC (56.5 ± 9.2%) was higher (p < 0.05) than all other supplementations. There were no differences in nuclear maturation or fertilization or in cleavage rates when comparing 1.5 mm NAC and 1.5 mm NACA supplementation to the control. Blastocyst formation for 1.5 mm NAC (44.4 ± 4.7%) and 1.5 mm NACA (46.2 ± 3.4%) supplementation were higher (p < 0.05) than the control (32.1 ± 6.2%) oocytes. These results indicate that supplementing 1.5 mm of NAC or NACA to the oocyte maturation medium increased the percentage of viable embryos reaching the blastocyst stage of development.     

Ascorbic Acid,Catalase and Chlorpromazine Reduce Cryopreservation‐induced Damages to Crossbred Bull Spermatozoaa

The present study evaluated the effectiveness of ascorbic acid, catalase, chlorpromazine and their combinations in reducing the cryodamages to crossbred bull (Bos taurus × Bos indicus) spermatozoa. A total of 32 ejaculates (eight each from four bulls) were diluted in Tris–citric acid–fructose–egg yolk–glycerol extender. Each ejaculate was split into six parts (five treatment and one control). Treatment groups included 10 mm ascorbic acid, 0.1 mm chlorpromazine, 200 IU/ml catalase, 10 mm ascorbic acid + 0.1 mm chlorpromazine or 200 IU/ml catalase + 0.1 mm chlorpromazine in the extender. Fluorescent probes (Fluorescein isothiocyanate–Pisum sativum agglutinin + Propidium iodide) were used for the assessment of spermatozoa viability and acrosomal status. The proportion of acrosome intact live (AIL), acrosome intact dead, acrosome reacted live and acrosome reacted dead sperm was assessed in fresh, equilibrated and frozen‐thawed semen. The functional status of the sperm was assessed using hypo‐osmotic sperm swelling test (HOSST). Activities of acrosin and hyaluronidase enzyme were also determined. Lipid peroxidation level was assayed based on the melonaldehyde (MDA) production. In cryopreserved semen, the values of AIL spermatozoa, HOSST response, hyaluronidase and acrosin activity were reduced by 53%, 47%, 34% and 54%, respectively from their initial values in fresh semen. However, MDA level was threefold higher in the frozen‐thawed sperm compared with fresh sperm. Significant (p < 0.05) improvement in motility, viability, HOSST response, retention of hyaluonidase and acrosin and reduction in MDA was recorded in ascorbic acid, catalase, ascorbic acid + chlorpromazine and catalase + chlorpromazine incorporated groups. The percentage of AIL sperm was significantly (p < 0.05) higher in ascorbic acid, catalase and ascorbic acid + chlorpromazine incorporated groups compared with the control. Chlorpromazine alone did not improve the post‐thaw semen quality but when combined with either ascorbic acid or catalse, improvement in semen quality was noticed. It was inferred that incorporation of ascorbic acid, catalase and ascorbic acid + chlorpromazine in semen extender improved the post‐thaw semen quality in crossbred bulls.     

Acceleration of Second and Fourth Metatarsal Fracture Healing with Recombinant Human Bone Morphogenetic Protein‐2/Calcium Phosphate Cement in Horses

Objective— To compare the efficacy of recombinant human bone morphogenetic protein‐2 (rhBMP‐2)/calcium phosphate (CP) to autogenous cancellous bone graft (CBG) and to no treatment on bone healing, in surgically induced osteotomies and ostectomies of the accessory metatarsal bones in an equine model. Study Design— Experimental. Animals— Adult horses (n=9). Methods— Segmental ostectomies of the second metatarsal bone (MT2) and osteotomies of the fourth metatarsal bone (MT4) were performed bilaterally in 9 horses. There were a total of 35 defects (1 MT4 was previously fractured) created and supplemented randomly either with no treatment (untreated control), rhBMP‐2/CP cement, or matrix (CPC or CPM), or CBG. Radiography was performed every 2 weeks until study endpoint at 12 weeks. After euthanasia, bone healing was evaluated using radiography, mechanical testing, and histology. Data were analyzed with ANOVA followed by the Duncan's Multiple Range Test or nonparametric analyses. Results— At 12 weeks, radiographic scores for union were significantly greater for the rhBMP‐2 (P<.0001) and CBG (P=.004) groups compared with the untreated control group, for both MT2 ostectomies and MT4 osteotomies. The rhBMP‐2 treated MT2 had greater maximum torque to failure in torsion than CBG and control limbs at 12 weeks (P=.011). Histologic analysis demonstrated increased bone formation and more mature bone at the ostectomy site for MT2 in the rhBMP‐2 and CBG groups compared with the untreated control group. Conclusion— Injection of rhBMP‐2/CP into surgically induced ostectomies and osteotomies of the accessory metatarsal bones might accelerate early bone healing in the horse. Clinical Relevance— RhBMP‐2/CP may be as effective if not superior to CBG as an adjuvant treatment to accelerate healing of bone defects.