Successful surgical management of eosinophilic granuloma on the urethral process of a gelding associated with Habronema spp. infection

Habronemiasis, otherwise known as summer sores, is a parasitic disease of horses, donkeys and other equids, commonly encountered in subtropical and tropical regions. It occurs as a result of nematode infestation of the skin and typically manifests as ulcerative granulating lesions or masses. Habronema muscae, Habronema microstoma and Draschia megastoma are the usual invaders, usually transmitted by house flies, face flies and stable flies. In this case report, an eosinophilic granuloma on the urethral process of a 15-year-old gelding is presented, followed by successful surgical excision without further medical treatment. To the authors’ knowledge, equine eosinophilic granuloma on the urethral process associated with habronemiasis has not previously been documented in southern Europe and should be considered as a potential differential diagnosis for penile masses.     

Cutaneous and ocular habronemiasis in horses: 63 cases (1988-2002)

OBJECTIVE: To describe clinical manifestations of cutaneous and ocular habronemiasis in horses and evaluate outcome of treatment. DESIGN: Retrospective study. ANIMALS: 63 horses. PROCEDURE: The diagnosis was made on the basis of history, clinical signs, and identification of calcified concretions (sulfur granules) in lesions. Histologic examination of biopsy specimens was used to confirm the diagnosis. Case horses were compared with a control population of 12,720 horses examined during the same period. RESULTS: Arabians, gray horses, and horses with diluted coat colors were overrepresented; Thoroughbreds were underrepresented. Lesions were identified most often during the summer and early fall. The medial canthus of the eye, male genitalia, third eyelid, and distal portions of the extremities were the most commonly affected locations. Twenty-five lesions were biopsied, and results of histologic examination were consistent with a diagnosis of habronemiasis. However, nematode larvae were seen in only 11 (44%) biopsy specimens. Treatment consisted of surgical removal (7 horses) or medical treatment (56) consisting of debulking granulation tissue and topical, intralesional, or systemic treatment with corticosteroids. All horses were treated with ivermectin. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that cutaneous and ocular habronemiasis should be considered when examining a horse during the summer months with a proliferative, moist, granulomatous lesion. Treatment should be aimed at decreasing the size of the lesion, reducing inflammation, and preventing recurrence. In general, the prognosis was good, and healing occurred within a few weeks. Fly control and regular deworming with ivermectin are recommended to reduce the incidence of habronemiasis.     

Chromomycosis in a horse

A 6-year-old Quarter Horse gelding was examined because it had a sharply circumscribed, firm, brown-black, roughly spherical dermal nodule at the right tuber ischii. After it was excised, the lesion did not recur. Microscopically, the dermis of the excised specimen had multiple pyogranulomas, many of which contained thick-walled, dark brown fungal elements, some with internal septation compatible with chromomycotic fungi. Chromomycosis is a rare skin disease in the horse. It may be included in the differential diagnoses of nodular and/or pigmented skin lesions that include melanoma, pyogranuloma, mycetoma, squamous cell carcinoma, habronemiasis, and onchocerciasis.     

Cutaneous habronemiasis in horses and domestic donkeys (Equus asinus asinus)

Cutaneous habronemiasis in 15 horses and 5 donkeys is described. The lesions were distributed in many parts of the body involving the medial canthus, shoulder and pectoral regions, knee and fetlock joints, abdominal wall and prepuce. Some animals had more than one lesion. The lesions were ulcerative and filled with soft light red granulation tissue. When curretted, the deeper layers revealed a dense fibrous tissue with calcified foci. Close examination of the lesions showed that the superficial layer of this dense fibrous tissue contained small caseated and necrotic foci. The same features prevailed in lesions involving the muscular areas, but the deeper layers consisted of a dense granuloma with no evidence of cicatrization. Curretted material digested in potassium hydroxide revealed fragments of larvae of the nematode suggestive of Draschia or Habronema. The histopathological changes were severe and comprised: necrotic foci in a dense fibrous stroma infiltrated with eosinophils, macrophages and few giant cells. Sections of the larvae surrounded by eosinophils were encountered in lesions of the muscular areas. Curretting and excision of the lesion were effective and produced complete healing of the wound by scar tissue formation.     

Full‐Thickness Skin Grafting to Cover Equine Wounds Caused by Laceration or Tumor Resection

Objective: To describe and evaluate full‐thickness skin grafting of equine wounds. Study Design: Case series. Animals: Adult horses (n=6). Methods: A full‐thickness graft was harvested from the pectoral region with the horse anesthetized or standing and sedated after local anesthetic infiltration. Grafts were attached to the cutaneous margin of the wound with staples and/or sutures if the horse was anesthetized or if the recipient site was desensitized. Cyanoacrylate glue was used to attach the grafts to the cutaneous margin of the wound of 3 horses. Medical records were reviewed for history, physical examination findings, grafting technique, postoperative complications, and outcome. Results: Three horses had full‐thickness skin grafting to cover a fresh defect created by excision of a cutaneous neoplasm, and 3 horses had full‐thickness skin grafting to cover a fresh or granulating laceration. Grafts were completely accepted in 5 horses. The superficial layers of all grafts sloughed, but the final cosmetic appearance of accepted grafts was good. Conclusions: Full‐thickness skin grafting can be performed in standing sedated horses with good cosmesis, especially when the meshed graft is expanded minimally. Clinical Relevance: Good acceptance of a full‐thickness graft can be expected, regardless of whether the graft is applied to a fresh or granulating wound.     

Co-occurrence of papillomas related to Equus caballus papillomavirus type 2 and cutaneous habronemiasis

The present report describes a case of cutaneous papillomatosis related to equine papillomavirus type 2 (EcPV2) with concomitant cutaneous habronemiasis. A 2-year-old female Pura Raza Española horse was referred for a dermatological evaluation because of the onset of cutaneous multifocal, nodular lesions on the face characterised by a warty and partially ulcerated overlying epidermis. Multiple biopsies were taken from these lesions, and cutaneous viral papillomas with a concomitant severe focal eosinophilic dermatitis with intralesional Habronematidae nematode larvae were diagnosed. Immunohistochemistry revealed papillomaviral capsid protein L1 in the epidermal proliferative lesion, confirming the presence of a productive infection. Rt-qPCR confirmed the presence of EcPV type 2. To the authors’ knowledge, this is the first report of EcPV2-related papillomas with concurrent habronemiasis.     

Nodular diseases

The development of a nodule is a fairly common reaction pattern in the skin and subcutaneous tissues of the horse. In many such nodules, the inciting aetiologic agent or antigen is unknown, but in some equine cases there is significant evidence for insect bite origin. The pathogenesis of some equine nodular diseases appears to involve marked collagen degeneration which varies from hyalinization to lysis and can become mineralized. The exact role the lytic collagen has in the pathogenesis of these lesions is still unknown. Vascular changes may be present in some entities. Other nodules may be caused by or complicated by the larva of Habronema spp. The clinical or histological entities outlined in this section include the collagenolytic granuloma, axillary nodular necrosis, unilateral papular dermatosis and cutaneous habronemiasis.     

Effects of hyperbaric oxygen on full-thickness meshed sheet skin grafts applied to fresh and granulating wounds in horses

OBJECTIVE: To determine the effects of hyperbaric oxygen therapy (HBOT) on full-thickness skin grafts applied to fresh and granulating wounds of horses. ANIMALS: 6 horses. PROCEDURES: On day 0, two 4-cm-diameter circular sections of full-thickness skin were removed from each of 2 randomly selected limbs of each horse, and two 4-cm-diameter circular skin grafts were harvested from the pectoral region. A skin graft was applied to 1 randomly selected wound on each limb, leaving the 2 nongrafted wounds to heal by second intention. On day 7, 2 grafts were harvested from the pectoral region and applied to the granulating wounds, and wounds grafted on day 0 were biopsied. On day 14, 1 wound was created on each of the 2 unwounded limbs, and the wounds that were grafted on day 7 were biopsied. All 4 ungrafted wounds (ie, 2 fresh wounds and 2 wounds with 1-week-old granulation beds) were grafted. The horses then received HBOT for 1 hour daily at 23 PSI for 7 days. On day 21, the grafts applied on day 14 were biopsied. RESULTS: Histologic examination of biopsy specimens revealed that grafts treated with HBOT developed less granulation tissue, edema, and neovascularization, but more inflammation. The superficial portion of the graft was also less viable than the superficial portion of those not treated with HBOT. CONCLUSIONS AND CLINICAL RELEVANCE: The use of HBOT after full-thickness skin grafting of uncompromised fresh and granulating wounds of horses is not indicated.     

Effects of split-thickness and full-thickness skin grafts on secondary graft contraction in horses.

Full-thickness, circular (4-cm diameter) cutaneous wounds were created on the metacarpi and metatarsi of 6 horses. Immediately after wounding, 1 wound on each horse received a meshed, split-thickness skin graft (0.64 mm) obtained from the ventrolateral aspect of the horse's thorax by use of a pneumatic dermatome, whereas a second wound received a meshed, full-thickness skin graft obtained from the pectoral area. In addition, sections of split-thickness and full-thickness grafts were refrigerated in a solution of McCoy's 5A medium, to which equine serum (10%) and gentamicin sulfate solution (16 mg/dl) were added. Ten days after wounding, 1 granulating wound on each horse was grafted with a stored, meshed, split-thickness graft, and 1 granulating wound on each horse was grafted with a stored, meshed, full-thickness graft. Areas of wounds were calculated from photographs taken of wounds on days 1, 5, 10, 15, 20, 25, and 30 after wounding. Time course of contraction was determined by use of a first-order mathematic model of changes in area through time. Rate constants of contraction for fresh or granulating wounds receiving full-thickness grafts did not differ significantly from those for fresh or granulating wounds receiving split-thickness grafts. Rate constants of contraction for grafted fresh wounds, however, were significantly less than those of grafted granulating wounds, regardless of whether a split-thickness or full-thickness graft was applied.     

Equine multisystemic eosinophilic epitheliotropic disease: A case report and review of literature

CASE HISTORY: A 2-year-old Standardbred gelding presented with a history of fever over 1 week, anorexia and skin lesions on all four legs. The lesions were associated with severe pruritus and oedema, and there was no response to therapy.

CLINICAL FINDINGS: The horse was in poor body condition, was lethargic and severely pruritic. Skin lesions consisted of diffuse alopecia and crusting of the distal extremities. Initially it was slightly febrile, but subsequently its temperature increased up to 40°C. Ten days after admission it developed profuse watery diarrhoea and the skin lesions progressed. Skin biopsies revealed superficial and deep perivascular dermatitis with lymphoplasmacytic and eosinophilic predominance. Based on the poor prognosis the horse was subject to euthanasia.

PATHOLOGICAL FINDINGS: The most notable lesions included ulcerative gastritis, typhlitis and colitis with prominent oedema of the intestines, marked subcutaneous oedema and severe thickening of the large bile ducts. Histopathology showed marked eosinophilic and lymphoplasmacytic infiltration of various tissues including the skin, gastrointestinal tract, mesenteric lymph nodes, large bile ducts, pancreatic duct and kidney. Immunohistochemistry revealed a clear predominance of CD3-positive cells in the lymphocytic infiltrations.

DIAGNOSIS: Based on the clinical findings and histopathology a diagnosis of multisystemic eosinophilic epitheliotropic disease (MEED) was made.

CLINICAL RELEVANCE: Multisystemic eosinophilic epitheliotropic disease is rare in horses, and usually chronic. In the current case the horse showed an apparently acute onset with high fever and rapid clinical deterioration. A diagnosis of MEED should be considered in horses presenting with weight loss and skin lesions with or without fever. A final diagnosis is based on histological results of biopsy specimens from affected organs.     

Diagnosis, relative incidence, and probable cause of cunean tendon bursitis-tarsitis of Standardbred horses

A prospective study of the diagnostic results on 25 previously untreated, slightly lame Standardbred horses showed that manipulative tests are of some help in diagnosis. The lameness of each horse was diminished or the horse went lame on the opposite limb after being given an injection of anesthetic in the cunean bursa, and lameness improved more when local anesthetic was injected in the distal intertarsal and tarsometatarsal articulations. In four horses, lesions of the distal articulation of the hock were evident on radiography.     

FEASIBILITY AND SAFETY OF CONTRAST‐ENHANCED ULTRASOUND IN THE DISTAL LIMB OF SIX HORSES

Vascular alterations play important roles in many orthopedic diseases such as osteoarthritis, tendonitis, and synovitis in both human and equine athletes. Understanding these alterations could enhance diagnosis, prognosis, and treatment. Contrast‐enhanced ultrasound (CEUS) could be a valuable method for evaluation of blood flow and perfusion of these processes in the equine distal limb, however no reports were found describing feasibility or safety of the technique. The goal of this prospective, experimental study was to describe the feasibility and safety of distal limb CEUS in a sample of six horses. For each horse, CEUS of the distal limb was performed after intravenous injections of 5 and 10 ml, as well as intra‐arterial injections of 0.5 and 1 ml contrast medium. Vital parameters were monitored and CEUS images were assessed qualitatively and quantitatively for degree of contrast enhancement. None of the horses had clinically significant changes in their vital parameters after contrast medium injection. One horse had a transient increase in respiratory rate, and several horses had mild increases of systolic blood pressure of short duration after intravenous, but not after intra‐arterial injections. Intra‐arterial injection was possible in all horses and resulted in significantly improved contrast enhancement both quantitatively (P = 0.027) and qualitatively (P = 0.019). Findings from this study indicated that CEUS is a feasible and safe diagnostic test for evaluation of the equine distal limb. Future studies are needed to assess the clinical utility of this test for horses with musculoskeletal diseases.     

The Bacteria Isolated from the Skin of 20 Horses at a Veterinary Teaching Hospital

The objective of this study was to isolate bacteria on the skin of the proximal to distal equine limb to guide the practitioner in the selection of prophylactic antimicrobial protocols. This prospective study involved 20 client-owned horses that were admitted to the Veterinary Teaching Hospital for routine elective surgery. Each horse spent between 12 and 36 hours at our hospital before sampling. Samples were collected from the skin of the left mid-thorax and the dorsal aspect of nine joints on the left side of each horse: front and hind coffin and fetlock joints, carpi, elbows, shoulders, hocks, and stifles. Samples were cultured aerobically and speciated when possible. When evaluating bacterial composition by location, a 40% difference was considered clinically significant. When comparing proximal sites above the fetlock to distal sites, the odds of isolating gram-positive bacteria were 1.23 times (P = .0124) higher at proximal sites; the odds of isolating coliform bacteria were 1.32 times (P = .023) higher at distal sites; and the odds of isolating a common septic arthritis pathogen were 1.16 times (P = .018) higher at distal sites. Coagulase-positive Staphylococcus was not isolated in this study. All comparisons between sites and between the proximal and distal limb were <40%, and thus were not considered clinically significant. No coagulase-positive Staphylococcus was isolated from any of the 200 sites in this study, suggesting that iatrogenic infections by that organism may not be because of preexisting flora. These data suggest that antibiotic prophylaxis targeting preexisting normal flora should be similar regardless of the area of interest on the limb.     

Multicentric mast cell tumors in a horse

Abstract: A 6‐year‐old female Rocky Mountain horse was presented for evaluation of draining tracts and distal limb subcutaneous edema on the left front and left hind limbs that had been present for 2 weeks. Direct smears of fluid collected by fine‐needle aspiration of subcutaneous fluid from both limbs were highly cellular with a predominance of eosinophils accompanied by numerous, moderately atypical, variably granulated mast cells. The cytologic diagnosis was mast cell tumor (MCT) with prominent eosinophilic infiltration with a differential diagnosis of eosinophilic granuloma. Histologic evaluation of surgical biopsies of lesions from both limbs was performed on sections stained with H&E, toluidine blue, and Luna stains. The histologic diagnosis was MCT, and staining with toluidine blue and Luna stains confirmed the presence of mast cells and eosinophils, respectively. In addition, the mast cells strongly expressed CD117. This is the first reported case of cutaneous mast cell neoplasia in a horse in which primary presenting complaints were draining tracts and distal limb subcutaneous edema involving multiple limbs. This case illustrates the utility of staining for CD117 expression in combination with traditional stains, such as toluidine blue and Luna, in differentiating MCTs from other eosinophilic lesions in horses.     

Computed tomographic evaluation of the distal limb in the standing sedated horse: Technique,imaging diagnoses,feasibility, and artifacts

In several veterinary institutions, adjustments of CT machines have been made that allow for imaging of the standing horse. The risk of general anesthesia is eliminated and the shorter scan completion time reduces cost to clients. The objective of this retrospective, analytical study was to evaluate the technique, imaging diagnoses, feasibility, and image artifacts of multi-slice helical CT of horses’ distal limbs acquired under standing sedation. The CT images of 250 horses of various breeds, aged 3–23 years, that underwent standing distal limb CT were evaluated. Three observers assessed the CT images for artifacts and inter-observer agreement was calculated. Eighty-six percent (95% confidence interval (CI), 81–90) of the scans were carried out on the forelimbs, while 14% (95% CI, 10–19) were of the hindlimbs. A total of 65% (95% CI, 59–71) of horses that underwent standing sedated CT had single imaging diagnoses. Seventy-one percent (95% CI, 65–77) of the cases had unilateral lesions, 27% (95% CI, 22–33) had bilateral lesions and 2% (95% CI, 1–4) had no diagnosed lesions. The average CT acquisition time was 17.5 minutes (range = 15–20). The average number of acquisitions per horse was 1.7 (median = 1; range = 1–4). There was good to excellent agreement between all three observers for the presence of motion artifact in the metacarpo/metatarsophalangeal joints, identification of marked beam hardening artifact, mild solar/ skin dirt, and photon starvation artifact (kappa 0.61-0.80). No complications were encountered. Standing examination of the distal limb achieved diagnostic image quality that was obtained with minimal acquisition attempts and in a timely manner.     

Laminin receptor 1 is differentially expressed in thoracic and limb wounds in the horse

Healing of wounds located on the distal limbs of horses is often complicated by retarded epithelialization and the development of exuberant granulation tissue (proud flesh). Treatments that definitively resolve this pathological process are still unavailable. Molecular studies of the repair mechanism might contribute to the development of new therapeutic strategies. The study presented herein aimed to clone the full length cDNA and to study the spatio-temporal expression profile of mRNA and protein for LAMR1, previously attributed a role in wound epithelialization, during the repair of body and limb wounds in the horse. Cloning was achieved by screening a cDNA library previously derived from 7-day wound biopsies. Expression was studied in unwounded skin and in samples from 1-, 2-, 3-, 4- and 6-week-old wounds of the body and limb. Temporal gene expression was determined by real time polymerase chain reaction (RT-PCR) while protein expression was mapped immunohistochemically. Full-length cDNA for equine LAMR1 was shown to be highly similar to that of other species. The mRNA expression of LAMR1 was significantly up-regulated only in thoracic wounds, 4 and 6 weeks following wounding (upon epithelialization). Cutaneous wounding induced protein expression at both locations. Our data suggest that up-regulation of LAMR1 protein might favour epithelialization during wound healing. However, its interaction with ligands other than laminin complicates data interpretation. Future studies should quantitatively verify the temporal expression of this protein in order to provide the basis for targeted therapies that might enhance epithelialization.     

OB‐cadherin cloning and expression in a model of wound repair in horses

Reason for performing study: Horses suffer from a debilitating impediment in repairing wounds located on the lower limb that leads to the development of a fibroproliferative disorder (exuberant granulation tissue). This condition is a source of wastage since it often forces retirement from competition. Treatments that resolve or prevent this condition are still lacking, maybe due to deficient knowledge of the underlying molecular mechanisms. Fibroblast‐to‐myofibroblast conversion is an essential step allowing contraction during wound repair and is accompanied by an increase in OB‐cadherin expression. Objectives: To clone equine cadherin‐11 (CDH11) cDNA and to study its spatiotemporal expression profile during the repair of body and limb wounds, thereby contributing to a better understanding of the repair process. Methods: Cloning was by a PCR technique. Expression was studied in intact skin and in 1, 2, 3, 4 and 6‐week‐old wounds of the body and limb. Temporal CDH11 gene expression was determined by RT‐PCR while OB‐cadherin protein expression was mapped immunohistochemically. Results: Equine CDH11 is a highly conserved gene and protein. mRNA was not expressed in equine skin whereas the wound repair process was characterised by a significantly higher expression in the thorax than in limb samples. mRNA expression pattern was paralleled by protein data as confirmed by immunohistochemistry. Conclusions: The data suggest that deficient OB‐cadherin expression in the first phases of wound repair contributes to the excessive proliferative response seen in horse limb wounds. Potential relevance: Future studies should verify the quantitative, temporal expression of this protein in order to provide the basis for targeted therapies that might prevent the development of EGT in horse wound repair.     

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.     

Scintigraphic evaluation of intra-arterial and intravenous regional limb perfusion of allogeneic bone marrow-derived mesenchymal stem cells in the normal equine distal limb using (99m) Tc-HMPAO

Reasons for performing study: Mesenchymal stem cells (MSCs) are commonly injected intralesionally for treatment of soft tissue injuries in the horse. Alternative routes of administration would be beneficial for treatment of lesions that cannot be accessed directly or to limit needle‐induced iatrogenic damage to the surrounding tissue. Objectives: The purpose of our study was to evaluate MSC distribution after intra‐arterial (IA) and intravenous (IV) regional limb perfusions (RLP) using scintigraphy. We hypothesised that MSCs would persist in the distal limb after tourniquet removal and that both techniques would lead to diffuse MSC distribution. Methods: Six horses were used in the study. MSCs were labelled with hexamethyl propylene amine oxime (HMPAO) and technetium‐99m. RLP was performed through the median artery of one forelimb and the cephalic vein of the opposite limb under general anaesthesia. The tourniquet was left in place for 45 min. Scintigraphic images were obtained at 0, 45, 75 min, 6 h and 24 h post injection. Results: Distribution of labelled MSCs through the entire distal limb was achieved with all 6 IA RLP, but 3 out of 6 IV RLP showed poor or absent uptake distal to the metacarpus. Mesenchymal stem cell persistence was 39% (30–60%) and 28% (14–50%) (median [minimum–maximum]) at 6 h for IA and IV RLP, respectively. Severe arterial thrombosis occurred in one horse after IA RLP. Conclusions: Both IA and IV RLP of the distal limb result in MSC persistence in perfused tissues. The IA perfusion resulted in more reliable cell distribution to the pastern and foot area. Potential relevance: Regional limb perfusion of MSCs might be used in cases where intralesional injection is not possible or in order to avoid iatrogenic needle damage. Further work is needed to assess the safety of IA RLP before its clinical use.     

Diagnosis and treatment of biaxial keratomas in the left fore foot of a 15‐year‐old Clydesdale mare

A 15‐year‐old Clydesdale mare presented for further diagnostics and treatment of waxing and waning lameness and recurrent subsolar abscesses. Radiographs and computed tomography revealed biaxial masses extending from the hoof capsule, causing bone resorption of the distal phalanx. Surgery was performed to remove the masses and post operative care included regional limb perfusions, systemic antibiotics and therapeutic shoeing. Histopathology was consistent with the diagnosis of keratoma for each of the masses; this is the first case of confirmed biaxial keratomas. Two months after surgery the horse is sound at the walk and is expected to return to full function within the next year.