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 This previously unpublished technique using a wire through the toe to create an active tension‐extension splint has a high success rate for correction of congenital flexural deformities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.     

Oxytetracycline associated acute kidney injury in a neonatal foal

A 3-day-old Australian Stock Horse filly presented with fulminant acute kidney injury after receiving 4 g of intravenous oxytetracycline at 24 and 48 h of age for medical management of flexural deformities. The filly’s clinical status initially improved, but worsening azotaemia prompted euthanasia after 55 h of supportive care. Necropsy confirmed proximal tubular necrosis. Nephropathy in association with oxytetracycline administration has been rarely reported. This case describes severe nephropathy in an apparently normal foal and highlights the need for thorough case evaluation prior to use of high-dose oxytetracycline therapy for correction of flexural deformities in foals.     

Effects of interleukin-1beta and tumor necrosis factor-alpha on expression of matrix-related genes by cultured equine articular chondrocytes

OBJECTIVE: To determine the effects of interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) on expression and regulation of several matrix-related genes by equine articular chondrocytes. SAMPLE POPULATION: Articular cartilage harvested from grossly normal joints of 8 foals, 6 yearling horses, and 8 adult horses. PROCEDURE: Chondrocytes maintained in suspension cultures were treated with various doses of human recombinant IL-1beta or TNF-alpha. Northern blots of total RNA from untreated and treated chondrocytes were probed with equine complementary DNA (cDNA) probes for cartilage matrix-related genes. Incorporation of 35S-sulfate, fluorography of 14C-proline labeled medium, zymography, and western blotting were used to confirm effects on protein synthesis. RESULTS: IL-1beta and TNF-alpha increased steady-state amounts of mRNA of matrix metalloproteinases 1, 3, and 13 by up to 100-fold. Amount of mRNA of tissue inhibitor of metalloproteinase-1 also increased but to a lesser extent (1.5- to 2-fold). Amounts of mRNA of type-II collagen and link protein were consistently decreased in a dose-dependent manner. Amount of aggrecan mRNA was decreased slightly; amounts of biglycan and decorin mRNA were minimally affected. CONCLUSIONS AND CLINICAL RELEVANCE: Treatment of cultured equine chondrocytes with IL-1beta or TNF-alpha resulted in marked alterations in expression of various matrix and matrix-related genes consistent with the implicated involvement of these genes in arthritis. Expression of matrix metalloproteinases was increased far more than expression of their putative endogenous inhibitor. Results support the suggestion that IL-1beta and TNF-alpha play a role in the degradation of articular cartilage in arthritis.     

Assessment of cartilage degradation effects of matrix metalloproteinase-13 in equine cartilage cocultured with synoviocytes

OBJECTIVE: To determine the effects of matrix metalloproteinase (MMP)-13, compared with interleukin (IL)-1alpha, on cartilage matrix molecule gene expression in a coculture system of equine cartilage explants and synoviocytes. SAMPLE POPULATION: Articular cartilage and synovium specimens harvested from femoropatellar joints of 4 horses, aged 3 to 5 years. PROCEDURES: Synoviocytes were isolated and cocultured with cartilage explants. Cultures were treated with human recombinant MMP-13 (1, 25, or 100 ng/mL) or IL-1alpha (0.01, 0.1, 1.0, or 10 ng/mL) for 96 hours, with medium exchange at 48 hours. Cartilage extracts and media were analyzed for glycosaminoglycan (GAG) content, and results were adjusted to cartilage DNA content. Quantitative PCR was performed on mRNA from cartilage (MMP-3, MMP-13, aggrecan, and collagen type IIB [COL2A1]) and synoviocytes (MMP-3 and MMP-13), and results were adjusted to 18S ribosomal subunit mRNA expression. Treatments were performed in triplicate, and the experiment was repeated 4 times. RESULTS: Cultures treated with MMP-13 or IL-1alpha had increased media GAG concentration at 48 and 96 hours. Aggrecan and COL2A1 mRNA expression were increased by application of MMP-13 or IL-1alpha. Gene expression of the catabolic mediator, MMP-3, in cartilage and synoviocytes was increased in cultures treated with MMP-13 or IL-1alpha. Expression of MMP-13 mRNA in cartilage was increased by IL-1alpha, but decreased in synoviocytes by MMP-13 treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Results support the use of recombinant MMP-13 in a coculture system of synoviocytes and cartilage explants for the study of osteoarthritis.     

Effects of platelet-derived growth factor-BB on the metabolic function and morphologic features of equine tendon in explant culture

OBJECTIVE: To evaluate the effects of recombinant human platelet-derived growth factor-BB (rhPDGF-BB) on the metabolic function and morphologic features of equine superficial digital flexor tendon (SDFT) in explant culture. Animals-6 euthanized horses (2 to 5 years old). METHODS: Forelimb SDFT explants were cultured for 6 days as untreated control specimens or treated with rhPDGF-BB (1, 10, 50, or 100 ng/mL of medium). Treatment effects on explant gene expression were evaluated via real-time PCR analysis of collagen type I, collagen type III, PDGF-A, and PDGF-B mRNA. Explants were assayed for total collagen, glycosaminoglycan, and DNA content; histologic changes were assessed via H and E staining and immunohistochemical localization of collagen types I and III. RESULTS: No morphologic or proliferative changes were detected in tendon explant sections. After high-dose rhPDGF-BB treatment, gene expression of collagen types I and III was increased and decreased, respectively. Expression of PDGF-A and PDGF-B mRNA was significantly increased at 24 hours, but later decreased to have few or negative autoinductive effects. Although PDGF gene expression waned after 48 hours of culture, collagen type I gene expression was significantly increased at 48 hours and reached peak value on day 6. Glycosaminoglycan and DNA content of explants were unchanged with rhPDGF-BB treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that rhPDGF-BB use may be of benefit in the repair of equine tendon, particularly through induction of collagen type I mRNA. Positive autoinductive effects of PDGF-BB in equine SDFT explants were detected early following culture medium supplementation, but these diminished with time.     

Long-term effects of desmotomy of the accessory ligament of the deep digital flexor muscle in standardbreds: 23 cases (1979-1989).

Desmotomy of the accessory ligament of the deep digital flexor muscle (inferior check desmotomy) permitted Standardbred foals affected with flexural deformities to reach their full athletic potential. Long-term effects of inferior check desmotomy were examined in 23 Standardbreds over a 10-year period. Six of 11 foals that were treated surgically either raced 6 times and obtained a race record or were training sound (if yearlings). All 12 horses with flexural deformity that did not receive an inferior check desmotomy had an unfavorable outcome (no race record). Foals that had surgery performed at a younger age apparently had a better chance of racing or training sound because no foals treated surgically after 8 months of age had a favorable outcome and only 1 foal that was older than 5 months at the time of surgery had a favorable outcome. In 5 foals that had surgery with an unsuccessful outcome, 3 were greater than or equal to 1 year old at the time of surgery and were lame when training was started on the limb(s) with the desmotomy.     

Myofibroblasts in the accessory ligament (distal check ligament) and the deep digital flexor tendon of foals

OBJECTIVE: To demonstrate myofibroblasts in the accessory ligament of the deep digital flexor tendon (ie, distal check ligament) and deep digital flexor tendon of clinically normal foals. SAMPLE POPULATION: Tissue specimens from 25 foals that were necropsied for reasons unrelated to this study and unrelated to musculoskeletal disease. PROCEDURE: The distal check ligament and deep digital flexor tendon of both forelimbs were examined histologically. Myofibroblasts were identified by immunohistochemical staining specific for alpha-smooth muscle actin (alpha-SMA). RESULTS: Most of the cells in the distal check ligament and deep digital flexor tendon of all foals stained positive for alpha-SMA. CONCLUSION AND CLINICAL RELEVANCE: Myofibroblasts made up most of the cells in the distal check ligament and deep digital flexor tendon of clinically normal foals. These cells have contractile ability and therefore, may play a role in flexure contracture of these tendons. The ability of tetracycline to chelate calcium or decrease the expression of the contractile protein alpha-smooth muscle actin could inhibit the myofibroblasts' ability to contract, thus providing a rationale for tetracycline administration as a treatment of distal interphalangeal joint flexor deformity in foals.     

Hypertrophy and physiological death of equine chondrocytes in vitro

REASON FOR PERFORMING STUDY: Equine osteochondrosis results from a failure of endochondral ossification during skeletal growth. Endochondral ossification involves chondrocyte proliferation, hypertrophy and death. Until recently no culture system was available to study these processes in equine chondrocytes. OBJECTIVE: To optimise an in vitro model in which equine chondrocytes can be induced to undergo hypertrophy and physiological death as seen in vivo. METHODS: Chondrocytes isolated from fetal or older (neonatal, growing and mature) horses were cultured as pellets in 10% fetal calf serum (FCS) or 10% horse serum (HS). The pellets were examined by light and electron microscopy. Total RNA was extracted from the pellets, and quantitative PCR carried out to investigate changes in expression of a number of genes regulating endochondral ossification. RESULTS: Chondrocytes from fetal foals, grown as pellets, underwent hypertrophy and died by a process morphologically similar to that seen in vivo. Chondrocytes from horses age >5 months did not undergo hypertrophy in pellet culture. They formed intramembranous inclusion bodies and the cultures included cells of osteoblastic appearance. Pellets from neonatal foals cultured in FCS resembled pellets from older horses, however pellets grown in HS underwent hypertrophy but contained inclusion bodies. Chondrocytes from fetal foals formed a typical cartilage-like tissue grossly and histologically, and expressed the cartilage markers collagen type II and aggrecan mRNA. Expression of Sox9, collagen type II, Runx2, matrix metalloproteinase-13 and connective tissue growth factor mRNA increased at different times in culture. Expression of fibroblast growth factor receptor-3 and vascular endothelial growth factor mRNA decreased with time in culture. CONCLUSIONS: Freshly isolated cells from fetal growth cartilage cultured as pellets provide optimal conditions for studying hypertrophy and death of equine chondrocytes. POTENTIAL RELEVANCE: This culture system should greatly assist laboratory studies aimed at elucidating the pathogenesis of osteochondrosis.     

Factors regulating collagen synthesis and degradation during second-intention healing of wounds in the thoracic region and the distal aspect of the forelimb of horses

OBJECTIVE: To determine significant molecular and cellular factors responsible for differences in second-intention healing in thoracic and metacarpal wounds of horses. ANIMALS: 6 adult mixed-breed horses. PROCEDURE: A full-thickness skin wound on the metacarpus and another such wound on the pectoral region were created, photographed, and measured, and tissue was harvested from these sites weekly for 4 weeks. Gene expression of type-I collagen, transforming growth factor (TGF)-beta1, matrix metalloproteinase (MMP)-1, and tissue inhibitor of metalloproteinase (TIMP)-1 were determined by quantitative in situ hybridization. Myofibroblasts were detected by immunohistochemical labeling with alpha-smooth muscle actin (alpha-SMA). Collagen accumulation was detected by use of picrosirius red staining. Tissue morphology was examined by use of H&E staining. RESULTS: Unlike thoracic wounds, forelimb wounds enlarged during the first 2 weeks. Myofibroblasts, detected by week 1, remained abundant with superior organization in thoracic wounds. Type-I collagen mRNA accumulated progressively in both wounds. More type-I collagen and TGF-beta1 mRNA were seen in forelimb wounds. Volume of MMP-1 mRNA decreased from day 0 in both wounds. By week 3, TIMP-1 mRNA concentration was greater in thoracic wounds. CONCLUSIONS AND CLINICAL RELEVANCE: Greater collagen synthesis in metacarpal than thoracic wounds was documented by increased concentrations of myofibroblasts, type-I collagen mRNA,TGF-beta1 mRNA, and decreased collagen degradation (ie, MMP-1). Imbalanced collagen synthesis and degradation likely correlate with development of exuberant granulation tissue, delaying healing in wounds of the distal portions of the limbs. Factors that inhibit collagen synthesis or stimulate collagenase may provide treatment options for horses with exuberant granulation tissue.     

Observations on the placentae of eight Thoroughbred foals born with flexural limb deformities

Congenital flexural deformities are relatively common in newborn Thoroughbred foals. This paper reports on the physical and morphological examination of 8 placentae from foals with varying degrees of the condition. All the placentae showed one or more of the following characteristics: a reduction in their linear dimensions; folding of the allantochorion over the course of major blood vessels; oedema. Although the changes observed in the placentae were indicative of uterine narrowing, it remains unclear if they are the cause of, an effect of or unrelated to the deformities.     

Effects of oral administration of concentrated equine serum IgG to newborn foals on passive immunity

Thirteen newborn foals of Quarter Horse breeding were used to determine if oral administration of concentrated equine serum increases concentrations of IgG in foals allowed to naturally suckle colostrum. Foals were alternately assigned either to receive 300 ml of an oral equine serum IgG product or to serve as controls. Foals receiving the IgG product were given 150 ml orally at 10 hours and again at 12 hours after birth. All foals were allowed to suckle from their dams ad libitum. Jugular blood samples were obtained from foals at 10 hours and 24 hours of age for IgG determination. Colostrum samples from the dam were also obtained within 3 hours following parturition for determination of specific gravity. Plasma samples were analyzed for IgG level using a commercially available radial immunodiffusion kit. Oral administration of equine serum IgG had no significant effect on concentrations of plasma IgG in foals at 24 hours of age (p>.34). There was also no difference between control and treated foals in the rate of IgG absorption from 10-24 hours after birth (p>.34). In conclusion, oral administration of equine IgG to foals that ingest their dam's colostrum does not significantly increase concentrations of plasma IgG when compared to controls.     

Detection and effects on platelet function of anti-platelet antibody in mule foals with experimentally induced neonatal alloimmune thrombocytopenia

Horse mares carrying mule foals were immunized during the last trimester of pregnancy with whole acid-citrate-dextrose-anticoagulated donkey blood to experimentally induce neonatal alloimmune thrombocytopenia. Thrombocytopenia occurred in the neonatal mule foals born to immunized horse mares within 24 hours after ingestion of their dams' colostrum. Mule foals born to mares not immunized with donkey blood did not develop thrombocytopenia. These findings suggest that antibodies may have been directed against a donkey platelet antigen present in the mule foals but not present in their dams. The objectives of this study were to determine whether anti-platelet antibody could be detected in mule foals with experimentally induced neonatal alloimmune thrombocytopenia, to identify any platelet proteins recognized by serum antibody in these foals, and to determine if platelet function was altered by sera from these mule foals. An indirect enzyme-linked immunosorbent assay demonstrated significantly higher absorption at 1:200 of platelet-bindable immunoglobulin G in serum from thrombocytopenic mule foals, compared with nonthrombocytopenic mule foals. Sera from thrombocytopenic and nonthrombocytopenic mule foals produced similar binding patterns in western immunoblots with donkey platelet proteins separated on sodium dodecyl sulfate polyacrylamide gels. Maximal platelet aggregation and relative slope of aggregation in response to collagen were significantly inhibited after incubation with sera from thrombocytopenic mule foals. These results suggest that mule foals with induced alloimmune thrombocytopenia have serum antibodies that bind to platelets and may compete with collagen binding sites to impair platelet aggregation.     

Effect of transforming growth factor beta1 on chondrogenic differentiation of cultured equine mesenchymal stem cells

OBJECTIVE: To determine the morphologic and phenotypic effects of transforming growth factor beta1 (TGFbeta1) on cultured equine mesenchymal stem cells (MSC) and articular chondrocytes. SAMPLE POPULATION: Bone marrow aspirates and articular cartilage samples from a 2-year-old and two 8-month-old horses. PROCEDURE: After initial isolation and culture, MSC and chondrocytes were cultured in Ham's F-12 medium supplemented with TGF-beta1 at a concentration of 0, 1, 5, or 10 ng/ml. Medium was exchanged on day 2, and cells were harvested on day 4. Medium was assayed for proteoglycan (PG) content. Total RNA was isolated from cell cultures, and expression of aggrecan, decrin, collagen type-I, and collagen type-II mRNA was assessed by means of Northern blot analyses. Cell cultures were stained with H&E or toluidine blue and examined histologically. Additional cultures were examined after immunohistochemical staining for type-I and -II collagen. RESULTS: MSC cultures exposed to TGF-beta1 had an increased cellular density with cell layering and nodule formation that was most pronounced in cultures treated with 5 ng of TGF-beta1/ml. Expression of collagen type-II mRNA in MSC cultures exposed to 5 ng of TGF-beta1/ml was 1.7 times expression in control cultures, and expression of collagen type-I mRNA was 2.8 times expression in control cultures. Treatment of MSC with TGF-beta1 led to dose-related increases in area and intensity of type-II collagen immunoreaction. CONCLUSION: Results suggest that TGF-beta1 enhances chondrogenic differentiation of bone marrow-derived MSC in a dose-dependent manner.     

Functional and ultrastructural changes in neutrophils from mares and foals experimentally inoculated with a respiratory tract strain of equine herpesvirus-1

Neutrophils isolated from venous blood of adult and foal ponies inoculated with equine herpesvirus-1 were evaluated by in vitro function tests and by electron microscopy. Foals had fever and severe neutropenia 24 hours after inoculation; increased neutrophil random migration under agarose and decreased antibody-dependent cell-mediated cytotoxicity were significant at 24 hours, but values had returned to preinoculation levels by 72 hours. Mares had fever and leukopenia of less severity, increases in neutrophil migration, and longer persistence of primary granule release than were seen in foals. Reduced migration and degranulation, and a decrease in antibody-dependent cell-mediated cytotoxicity seen with neutrophils from foals, as compared with mares, may relate to the high susceptibility of foals to equine herpesvirus-1 infection.     

Pharmacokinetics of oxytetracycline in clinical cases in the red-necked wallaby (Macropus rufogriseus)

Oxytetracycline, administered at 40 mg kg-1 by intravenous injection, was used as part of the treatment of three female red-necked wallabies, Macropus rufogriseus, with suspected Fusobacterium necrophorum infections. The plasma concentrations of this drug in blood samples collected at intervals up to 24 hours after administration were measured using a biological assay. The pattern of decline in plasma oxytetracycline concentration with time was consistent with a two-compartment model. The half-life of the elimination phase was calculated to be 11.4 hours and the apparent volume of distribution was found to be 2.041 litres kg-1. These results provide a basis for devising appropriate oxytetracycline dosage regimes for the species.     

Absorption of bovine colostral immunoglobulins G and M in newborn foals

The uptake of colostral IgG and IgM, their serum half-lives, and the rates of endogenous synthesis of IgG and IgM were evaluated in 6 newborn foals fed bovine colostrum (principals) and 6 foals allowed to suckle their dams (controls). The principal foals were fed 400 ml of bovine colostrum (IgG, 10,000 mg/dl and IgM, 200 mg/dl) at 2-hour intervals, from 2 to 20 hours after foaling (total dose, 4 L). Serum IgG and IgM concentrations were determined by single radial immunodiffusion from birth to 98 days of age. At foaling, principal foals had no detectable serum equine IgG, but 1 control foal had serum equine IgG of 185 mg/dl. After ingestion of colostrum, there was no significant difference in the maximal serum bovine IgG concentration (range, 1,350 to 3,300 mg/dl) in the principal foals, and maximal serum equine IgG concentration in the control foals (range, 500 to 6,000 mg/dl). The calculated biological bovine and equine IgG half-life in the principal and control groups was 9.4 and 26 days, respectively. Endogenous IgG synthesis was first detected in 1 principal foal at 3 days of age, but was detected first between 28 and 42 days in the other principal foals. Starting on day 56 there was no significant difference in serum equine IgG concentration between groups. At foaling, foals in both groups had low equine IgM concentrations. In the control foals, there was marked individual variation in the increases in equine IgM concentration (range, 5 to 73 mg/dl) after ingestion of colostrum.(ABSTRACT TRUNCATED AT 250 WORDS)     

Farriery for the foal: A review part 2: Therapeutic farriery

The extensive nature of this topic warrants this review paper to be divided into two parts: ‘Basic trimming in foals’ and ‘Therapeutic farriery in foals’. Management of the feet and limbs during this juvenile period will often dictate the success of the foal as a sales yearling or mature sound athlete. Overall hoof care in the foal is often a joint venture between the veterinarian and the farrier. The orthopaedic disorders discussed in this paper that require input from the two professions are flexural limb deformities (FLD) and angular limb deformities (ALD). The concept of protecting the foot from the deleterious effects of mal-loading created by many FLDs and ALDs is just as important as using the symptomatology as an instrument to correct the deformity. This paper presents a review of the current information regarding the farriery for these two limb deformities while dispelling some of the anecdotal methodology, such as the use of toe extensions to treat flexural deformities, that presently exists. Considering the deficiency of information in the literature, segments of this text will be based on the author's extensive clinical practice, comprehensive clinical records and comparisons of case outcomes.     

Equine Herpesvirus 1 Infection in Mares Vaccinated with a Live-virus Rhinopneumonitis Vaccine Attenuated in Cell Culture

Vaccination, in July and again in either November or December 1976, of 55 pregnant Standardbred mares with a live-virus rhinopneumonitis vaccine attenuated in cell culture failed to protect some mares from infection with equine herpesvirus 1. From 1976-12-08 to 1977-03-08, 33 mares foaled healthy foals, 16 mares foaled dead foals or live foals which died usually within 48 hours and six mares aborted. Gross and histological examinations and virus isolation studies confirmed that equine herpesvirus 1 caused 18 of the 22 neonatal deaths, stillbirths or abortions.     

Some features of the antibody response of sheep

Six nine-month-old red deer were injected intramuscularly with a long-acting injectable solution of oxytetracycline (“Terramycin-LA”, Pfizer Ltd) at a dose rate of 20 mg/kg. Four similar control deer were injected with saline. There was no significant pain response to injection, and only minor palpable swellings at the injection site were observed in three oxytetracycline-treated and one control animal. No statistically significant changes in white blood cell numbers, blood fibrinogen, creatine Phosphokinase or glutamic oxaloacetic transaminase concentrations occurred as a result of Oxytetracycline administration up to seven days after injection. Mean plasma Oxytetracycline concentration reached a peak (4.68 mg/1) two hours after injection and declined to levels below assay sensitivity (0.3 mg/1) in five deer 72 hours after injection, and in all deer by 96 hours after injection. No gross lesions at the injection site were observed at slaughter 30 days after injection. There were traces of Oxytetracycline at the injection site muscle of two deer after 30 days, but residues were not detected in injection site muscle from the other four deer, or in any of the liver, kidney or other muscle specimens.