Clinical outcome after surgical correction of cleft palate by laryngeal tie-forward in 2 horses

Introduction: Cleft palate is a rare congenital defect in horses. Its description in literature is generally limited to cases of young foals referred for milk regurgitation from nostrils, coughing and dysphagia, with a guarded to poor prognosis if not treated. Surgical intervention consists mainly in palatoplasty that is considered a salvage procedure and complications are frequent. Only few cases of cleft palate in adult horses are described in literature. Laryngeal tie-forward, a surgical technique generally used to treat dorsal displacement of the soft palate, has been described only in one 4-year-old pony with cleft palate but clinical improvement after surgery was partial and temporary. Objectives: To describe laryngeal tie-forward as a valid option for the treatment of cleft palate in two adult horses not suitable for palatoplasty. Methods: Clinical records of two 8 y.o. jumping horses performing respectively at a low and intermediate intensity of exercise and competition. Both horses were referred for respiratory noise, exercise intolerance, cough and alimentary nasal discharge; they were diagnosed with a moderate soft palate defect and surgically treated by laryngeal tie-forward. A clinical examination and control endoscopy were realized at 4 months for one horse and at 3 years after surgery for the other. Results: In both horses postoperative endoscopy showed a visible reduction of the gap between soft palate and the larynx so that the epiglottis was covering the soft palate defect. One horse was examined 4 months after surgery, a residual respiratory noise during exercise was still present but lighter than prior to intervention, its performances had remarkably improved. The second horse was controlled 3 years after surgery, some cough was still observed but limited to the onset of exercise, performances were satisfying and the horse was working at the same intensity level. Globally, in both horses, initial symptoms had disappeared or were significantly reduced after surgery and owners were satisfied. Reduction of the soft palate defect was still effective, even several months after surgery. Conclusions: The description of these two clinical cases shows that some horses can grow up to adult age with moderate defects of the soft palate; they are likely to show clinical signs of variable severity. In such cases laryngeal tie-forward should be considered as an option if the defect is too large or asymmetrical to be corrected with palatoplasty

Generalized sarcoidosis and hypertrophic osteopathy in a Standardbred with poor-performance.

editorial reviewedBackground and objective: Equine sarcoidosis (ES) is a rare but emergent disease of unknown etiology. Body of literature on ES remains limited, especially on the generalized forms associated with hypertrophic osteopathy (HO). This case report describes clinical and imaging findings in a racehorse with exercise intolerance affected by generalized ES and subsequent HO. Material and methods: A 6-year-old Standardbred gelding was presented for sudden exercise intolerance, weight loss, pyrexia, skin lesions, facial and peripheral multifocal swelling. Results: The horse underwent diagnostic imaging of head and limbs. Radiography and ultrasonography of the head revealed periostitis and regional soft tissue swelling of the maxillary bone. Bilateral marked periostitis with palisade-like appearance involving diaphyseal or metaphyseal region was also identified on distal radius and metatarsal bones, suggestive of HO. Computed tomography excluded dental or sinus abnormalities. Due to the appearance (scaling, crusting and exfoliating) of skin lesions on the croup and lower hindlimbs during hospitalization, biopsies were performed revealing granulomatous dermatitis. This finding was consistent with ES, thus explaining HO. Rapidly declining conditions of the horse led to the decision of euthanasia. Post-mortem examination revealed splenomegaly and several pulmonary nodules. Histopathology on different organs confirmed severe granulomatous lesions; PCR on a pool of tissues identified EHV-5. Discussion and conclusions: ES, even in its generalized form, is an emergent disease that should be considered in case of unspecific clinical signs combining exercise intolerance, weight loss, facial/distal limb bone deformities and skin lesions; EHV-5 could play a role in the development of the disease

A case of true hermaphrodism in a horse

peer reviewedTrue hermaphroditism in horses, is a complex and poorly under- stood disorder of sexual development characterized by the presence of both male and female gonads as separate organs or in a combined structure called ovotestis. A 3-year-old Spanish nullipa- rous mare with a history of stallion behaviour and an abnormal reproductive exam was presented at the Equine Clinic of the University of Li ege. Externally, a small vulva with an enlarged clitoris was observed. Transrectal ultrasonography showed a thin (<1 cm) uterus, leading to a small heterogeneous structure in the normal place of the left ovary. No structure could be identified in the presumed location of the right ovary. Speculum exam was compatible with a vaginal agenesis, as the cervix opened craniad the urethral meatus. Oestradiol, progesterone and testosterone concen- trations were compatible with a stallion’s endocrinology. Gonads were laparoscopically found in the usual location of the ovaries in both flanks and excised. Histopathology of both gonads showed atrophic testicular tissue with hyperplastic Leydig cells. The left gonad also contained ovarian tissue with some scarce primordial follicles. Clitoral enlargement is the first symptom most commonly identified in animals with true hermaphroditism and can be explained by the production of testosterone by the gonads, however, the aetiology of the vaginal agenesis is unknown. True hermaph- roditism in horses is generally related to 64XX syndrome with or without SRY gene translocation (impending karyotyping analysis). After 5 days the mare was discharged from the clinic, and in absence of testosterone the stallion behaviour disappeared in a couple of months

Presence of neutrophil myeloperoxidase in lamellar tissue of horses with laminitis induced using a hyperinsulinemic model

peer reviewedLaminitis is a pathology of the equine digit resulting in failure of the dermo-epidermal interface. Inflammation is likely a central player in its pathophysiology. Neutrophil activation and the presence of myeloperoxidase have been observed in the black walnut heartwood extract model, used to investigate sepsis-related laminitis. However, less is known about the role of neutrophil activation in the prolonged euglycemic hyperinsulinemic clamp (pEHC) model. This model is used to investigate the pathophysiology of laminitis related to endocrine disturbances like hyperinsulinemia encountered in pituitary pars intermedia dysfunction and equine metabolic syndrome. Five horses were included in this study and randomly assigned to a control group (n = 2) and a pEHC group (n = 3). Histological sections of lamellar tissue from all horses were obtained and immunohistochemically stained for myeloperoxidase and counterstained with hematoxylin-eosin. The sections were examined for histopathological evidence of laminitis and myeloperoxidase. Histopathological changes that characterize insulin-induced laminitis and increased presence of myeloperoxidase, especially in the dermal lamellae, were observed in histologic sections of pEHC-treated horses compared to control horses. Neutrophil activation with myeloperoxidase release may play a role in the pathophysiology of endocrinopathic laminitis. The implications of this finding warrant further investigation

Gastrointestinal effects of general anaesthesia in horses undergoing non abdominal surgery: focus on the clinical parameters and ultrasonographic images

The ultrasonographic images of the gastrointestinal tract in horses can be influenced by fasting and sedation but the proper effect of general anaesthesia (GA) on them has not been determined yet. This study aimed to evaluate the effects of GA on ultrasonographic images of the gastrointestinal tract in horses and to compare these effects with a clinical evaluation. Twenty horses undergoing non-abdominal surgeries were evaluated by ultrasonography before and 4 times within 24 h after GA. Each ultrasonographic exam focused on the stomach, the duodenum and on 5 locations on the jejunum. The four-quadrant auscultation and the postoperative faecal output were also recorded. Pre and post anaesthetic values were compared using linear mixed effects models. None of the horses presented colic signs or reduced faecal output. During the first 2 post anaesthetic evaluations, the gut sounds were significantly decreased and, when taking all jejunal locations together, the jejunal diameter and visualisation frequency significantly increased. No intestinal loop appeared thickened and most of their diameters remained within the normal range. Our results suggest that the effects of GA on the ultrasonographic images of the small intestine are mild and of short duration and can therefore be differentiated from a pathological process

Effects of Juglone on Neutrophil Degranulation and Myeloperoxidase Activity Related to Equine Laminitis.

Experimental laminitis, characterized by a failure of the dermal-epidermal interface of the foot, can be induced in horses by the oral administration of a black walnut extract (BWE). In the early phase of this severe and painful disease, an activation of neutrophil occurs, with the release of myeloperoxidase (MPO), a pro-oxidant enzyme of neutrophils, in plasma, skin, and laminar tissue. Juglone, a naphthoquinone derivative endowed with redox properties, is found in walnuts and has been incriminated in this neutrophil activation. We report for the first time the inhibitory activity of juglone on the degranulation of neutrophils induced by cytochalasin B and formyl-methionyl-leucyl-phenylalanine as monitored by the MPO release (>90% inhibition for 25 and 50 μM). Moreover, it also acts on the peroxidase activity of MPO by interacting with the intermediate "π cation radical," as evidenced by the classical and specific immunological extraction followed by enzymatic detection (SIEFED) assays. These results are confirmed by a docking study showing the perfect positioning of juglone in the MPO enzyme active site and its interaction with one of the amino acids (Arg-239) of MPO apoprotein. By chemiluminescence and electron paramagnetic resonance techniques, we demonstrated that juglone inhibited reactive oxygen species (ROS) and superoxide anion free radical produced from phorbol myristate acetate (PMA)-activated polymorphonuclear neutrophils (PMNs). These results indicate that juglone is not the trigger for equine laminitis, at least if we focus on the modulation of neutrophil activation

Effect of non-sperm cells removal with single layer colloidal centrifugation on myeloperoxidase concentration in post-thaw equine semen

Myeloperoxidase (MPO) is a pro-oxidant enzyme contained in, and released by, neutrophils during degranulation or after lysis. Post-thaw semen contains MPO and its concentration is associated with decreased sperm motility. Recently, MPO concentration in post-thaw semen was shown to be associated with presence of non-sperm cells. The aim of this study was to describe the effect of centrifugal fractionation of semen prior to cryopreservation on post-thaw concentrations of non-sperm cells and MPO. The experimental design consisted in freezing semen with or without previous centrifugation through two concentrations of single layer colloid media. Non-sperm cells and MPO concentrations were assessed in pellet and upper layer at each step of the procedure and MPO was detected in cells by immunocytochemistry. Single layer colloid centrifugation decreased non-sperm cells and MPO concentrations in post-thaw semen. The MPO concentration was correlated with concentration of non-sperm cells in the upper layer of the supernatant. In post-thaw semen, with or without previous single layer colloid centrifugation, MPO concentration was correlated with concentration of non-sperm cells. Overall, neutrophils were rarely observed and non-sperm cells were mainly epithelial cells or cellular debris, as demonstrated by MPO immunocytochemistry. Following single layer colloid centrifugation, MPO concentration was decreased, and correlations observed in different samples of the experiments revealed an association between concentrations of MPO and non-sperm cells. At all steps of the semen processing and cryopreservation, MPO immunostaining was clearly identified only on non-sperm cells. Our results demonstrate that non-sperm cells present in fresh semen release MPO during freezing.SPERMP

Constitutive apoptosis in equine peripheral blood neutrophils in vitro

AbstractThe aim of this study was to characterise constitutive apoptosis in equine peripheral blood neutrophils, including assessment of factors that potentially modulate neutrophil survival through alteration of the rate of constitutive apoptosis. Cells underwent spontaneous time-dependent constitutive apoptosis when aged in culture for up to 36 h, developing the structural and functional features of apoptosis observed in many cell types, including human neutrophils. Neutrophils undergoing apoptosis also had diminished zymosan activated serum (ZAS)-stimulated chemiluminescence, but maintained responsiveness to phorbol myristate acetate (PMA). The constitutive rate of equine neutrophil apoptosis was promoted by lipopolysaccharide (LPS), tumour necrosis factor α and phagocytosis of opsonised ovine erythrocytes, while it was inhibited by dexamethasone and ZAS (a source of C5a). Formyl-Met-Leu-Phe, leukotriene B4, platelet activating factor and PMA had no demonstrable effect on equine neutrophil apoptosis. There was a difference between equine and human neutrophil apoptosis in response to LPS and the time-dependence of the response to dexamethasone

Polyphenol Content and Modulatory Activities of Some Tropical Dietary Plant Extracts on the Oxidant Activities of Neutrophils and Myeloperoxidase

Young leaves of Manihot esculenta Crantz (Euphorbiaceae), Abelmoschus esculentus (Malvaceae), Hibiscus acetosella (Malvaceae) and Pteridium aquilinum (Dennstaedtiaceae) are currently consumed as green vegetables by peoples in sub-Saharan Africa, Latin America, Asia and their migrants living in Western Europe. Sub-Saharan peoples use Manihot, Abelmoschus and Hibiscus also in the folk medicine to alleviate fever and pain, in the treatment of conjunctivitis, rheumatism, hemorrhoid, abscesses, ... The present study investigates the effects of aqueous extracts of those plants on the production of reactive oxygen species (ROS) and the release of myeloperoxidase (MPO) by equine neutrophils activated with phorbol 12-myristate 13-acetate (PMA). The ROS production was measured by lucigenin-enhanced chemiluminescence (CL), and the release of total MPO by an ELISA method. The study also investigates the effect of the extracts on the activity of MPO by studying its nitration activity on tyrosine and by using a new technique called SIEFED (Specific Immunological Extraction Followed by Enzymatic Detection) that allows studying the direct interaction of compounds with the enzyme. In all experiments, the aqueous extracts of the plants developed concentration-dependent inhibitory effects. A moderate heat treatment did not significantly modify the inhibitory capacity of the extracts in comparison to not heated ones. Total polyphenol and flavonoid contents were determined with an HPLC-UV/DAD analysis and a spectroscopic method using Folin-Ciocalteu reagent. Some polyphenols with well-known antioxidant activities (caffeic acid, chlorogenic acid, hyperoside, rosmarinic acid and rutin) were found in the extracts and may partly explain the inhibitory activities observed. The role of those dietary and medicinal plants in the treatment of ROS-dependent inflammatory diseases could have new considerations for health