There are morphological and reproductive physiological differences between swamp buffalo (Bubalus carabanensis) and river buffalo (Bubalus bubalis). The development of fetus weight and fetus biometry was reported in river buffalo and other animals but not in swamp buffalo. The aim of this study was to describe the inherent variability in fetus related measurements during swamp buffalo pregnancy. The data is based on measurements of 267 fetuses and 5 new born claves from swamp buffalo. The results show that a significant linear correlation exists between estimated age of fetuses and parameters of fetus sizes. There were correlations between crown-rump length (CRL) and other fetal parameters, as well as between fetus weight and its parameters. In conclusion, our data indicated that the feasibility and value of fetal measures in swamp buffaloes being used for the evaluation of fetal development.

peer reviewed | The aim of this work was to measure the myeloperoxidase (MPO) concentration in bronchoalveolar lavage (BAL) fluid collected from horses with recurrent airway obstruction (RAO), both in crisis and in remission, as well as from healthy horses. Seven horses with RAO were exposed to moldy hay until the maximum change in pleural pressure was greater than 1.5 kPa. At that point, BAL was performed, and the total cell counts and percentages in the fluid were immediately determined. To measure the MPO concentration in BAL-fluid supernatant, we used a specific enzyme-linked immunosorbent assay with polyclonal antibodies against equine MPO. The tests were repeated on the horses with RAO after they had spent 2 mo on pasture. Six healthy horses serving as controls underwent the same tests. The absolute and relative neutrophil counts and the MPO concentration in the BAL fluid were significantly greater in the horses with an RAO crisis than in the control horses. After 2 mo on pasture, the horses that had been in RAO crisis were clinically normal, and their neutrophil counts and MPO levels in BAL fluid had significantly decreased; during remission their neutrophil counts were not significantly different from those in the healthy horses, but their MPO concentration remained significantly higher. This study showed that determining the MPO concentration in a horse's BAL fluid is technically possible and that during remission from RAO the concentration remains higher than normal. Thus, MPO may be a marker of neutrophil presence and activation in the lower airways. | Le but de cette étude était de mesurer la concentration de myéloperoxydase (MPO) dans le liquide de lavage broncho-alvéolaire (BAL) récolté chez des chevaux avec obstruction respiratoire récurrente (RAO), en période de crise et en rémission, ainsi que chez des chevaux en santé. Sept chevaux avec RAO ont été exposés à du foin moisi jusqu’à ce que le changement maximal dans la pression pleurale soit plus grand que 1,5 kPa. À cet instant on procéda au BAL et le dénombrement cellulaire total de même que les pourcentages dans le liquide furent immédiatement déterminés. Afin de mesurer la concentration de MPO dans le liquide de BAL on procéda à une épreuve immunoenzymatique spécifique utilisant des anticorps polyclonaux dirigés contre MPO. Les épreuves ont été répétées chez les chevaux avec RAO après qu’ils eurent passé 2 mois au pâturage. Six chevaux en santé servant de témoin ont subi les mêmes tests. Les comptes absolus et relatifs de neutrophiles et les concentrations de MPO dans le liquide de BAL étaient significativement plus élevés chez les chevaux avec RAO en période de crise que chez les chevaux témoins. Après 2 mois au pâturage, les chevaux qui avaient été en crise de RAO étaient cliniquement normaux et leurs comptes en neutrophiles et niveaux de MPO dans le liquide de BAL avaient diminué de manière significative; durant la rémission les comptes de neutrophiles n’étaient pas significativement différents de ceux des chevaux témoins, mais les concentrations de MPO sont demeurées significativement plus élevées. Cette étude a permis de démontrer qu’il était techniquement possible de déterminer la concentration de MPO dans du liquide de BAL équin et que durant la période de rémission d’une crise de RAO la concentration est demeurée plus élevée que la normale. Ainsi, la MPO pourrait être un marqueur de la présence et de l’activation des neutrophiles dans les voies respiratoires inférieures. (Traduit par Docteur Serge Messier)

peer reviewed | [en] OBJECTIVE: To determine and compare the concentration of gentamicin in the lower airways and serum of healthy spontaneously breathing dogs after nebulization with 5% undiluted gentamicin during 3 versus 10 minutes. ANIMALS: 10 healthy experimental Beagles. METHODS: This was a prospective crossover study. A standardized bronchoalveolar lavage (BAL) procedure was performed in each dog after 1 week of administration of each of 2 different gentamicin nebulization protocols separated by a 1-week washout period. The 2 protocols consisted of nebulization of 5% undiluted gentamicin (50 mg/mL) twice daily either during 10 minutes per session (± 95 mg; 10-minute protocol) or 3 minutes per session (± 28 mg; 3-minute protocol). BAL fluid (BALF) was obtained under general anesthesia using a bronchoscope within 15 minutes after administration of the last nebulization. Blood was collected within 5 minutes after BALF collection. BALF and serum gentamicin concentrations were determined by particle-enhanced turbidimetric inhibition immunoassay. Concentrations between protocols were compared using a paired t test. RESULTS: Both BALF and serum gentamicin concentrations were higher after the 10-minute protocol compared with the 3-minute protocol (mean ± SD: 2.41 ± 0.87 mg/L vs 1.25 ± 0.31 mg/L, P = .001; and 1.02 ± 0.59 mg/L vs 0.31 ± 0.24 mg/L, P < .0001 in BALF and serum, respectively), while the BALF-to-serum ratio did not differ between the protocols (3.75 [1.37 to 5.75] (median [IQR]) in the 3-minute protocol vs 2.48 [2.02 to 2.67] in the 10-minute protocol; P = .754). CLINICAL RELEVANCE: A 3-minute nebulization of gentamicin seems to achieve sufficient concentrations of gentamicin in the BALF to have good efficacy against aminoglycoside-sensitive bacteria while remaining below the toxic range values in blood.

peer reviewed | OBJECTIVE: To compare the extent of inflammation and catabolic collagen response in the middle carpal joints (MCJs) of healthy horses following intra-articular injection of 2% lidocaine, 2% mepivacaine, lactated Ringer solution (LRS), or 0.1% methyl parahydroxybenzoate. ANIMALS: 17 adult horses. PROCEDURES: In the first of 2 experiments, the left middle carpal joint (MCJ) of each of 12 horses was injected with 10 mL of 2% lidocaine (n = 3), 2% mepivacaine (3), or LRS (control; 6). After a 4-week washout period, the right MCJ of the horses that received lidocaine or mepivacaine was injected with 10 mL of LRS, and the right MCJ of horses that received LRS was injected with 10 mL of 2% lidocaine (n = 3) or 2% mepivacaine (3). In experiment 2, the left MCJ of each of 5 horses was injected with 10 mL of 0.1% methyl parahydroxybenzoate. After a 48-hour washout period, the right MCJ of each horse was injected with 10 mL of LRS. Synovial fluid (SF) samples were aseptically collected before and at predetermined times after each injection. Synovial fluid WBC count, neutrophil percentage, and total protein, neutrophil myeloperoxidase, neutrophil elastase, and Coll2-1 concentrations were compared among treatments. RESULTS: Both lidocaine and mepivacaine induced SF changes indicative of inflammation and a catabolic collagen response, but the magnitude of those changes was more pronounced for lidocaine. Methyl parahydroxybenzoate did not cause any SF changes indicative of inflammation. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that mepivacaine was safer than lidocaine for intra-articular injection in horses.

peer reviewed | Objective—To culture equine myoblasts from muscle microbiopsy specimens, examine myoblast production of reactive oxygen species (ROS) in conditions of anoxia followed by reoxygenation, and assess the effects of horseradish peroxidase (HRP) and myeloperoxidase (MPO) on ROS production. Animals—5 healthy horses (5 to 15 years old). Procedures—Equine skeletal myoblast cultures were derived from 1 or 2 microbiopsy specimens obtained from a triceps brachii muscle of each horse. Cultured myoblasts were exposed to conditions of anoxia followed by reoxygenation or to conditions of normoxia (control cells). Cell production of ROS in the presence or absence of HRP or MPO was assessed by use of a gas chromatography method, after which cells were treated with a 3,3′-diaminobenzidine chromogen solution to detect peroxidase binding. Results—Equine skeletal myoblasts were successfully cultured from microbiopsy specimens. In response to anoxia and reoxygenation, ROS production of myoblasts increased by 71%, compared with that of control cells. When experiments were performed in the presence of HRP or MPO, ROS production in myoblasts exposed to anoxia and reoxygenation was increased by 228% and 183%, respectively, compared with findings for control cells. Chromogen reaction revealed a close adherence of peroxidases to cells, even after several washes. Conclusions and Clinical Relevance—Results indicated that equine skeletal myoblast cultures can be generated from muscle microbiopsy specimens. Anoxia-reoxygenation– treated myoblasts produced ROS, and production was enhanced in the presence of peroxidases. This experimental model could be used to study the damaging effect of exercise on muscles in athletic horses.

peer reviewed | Gastrointestinal disorders, especially strangulating intestinal obstructions, are still a major cause of illness and death in the horse. Circulating lipopolysaccharides may activate both neutrophils and monocytes. The activated neutrophils release myeloperoxidase (MPO), a specific enzyme with strong oxidative activity. The aim of this study was to evaluate MPO concentrations in the plasma and peritoneal fluid (PF) of horses with colic and to check the hypothesis that these concentrations would be higher in a case of strangulating obstruction than in cases of nonstrangulating disease. By using a specific enzyme-linked immunosorbent assay for equine MPO, we determined the MPO concentrations in horses admitted to a clinic for colic. Horses with nonstrangulating or strangulating obstruction of the large intestine (NSLI or SLI), strangulating obstruction of the small intestine (SSI), or inflammatory bowel disease (IBD) were compared with healthy horses. The horses with SLI, SSI, or IBD had significantly higher MPO levels in plasma and PF than did those in the other 2 groups. The mean plasma level was significantly higher in the horses with NSLI than in the healthy horses. High MPO values in PF indicated necrotic bowel. These results show that neutrophil activation occurs during nonstrangulating and strangulating intestinal obstruction in horses and that the plasma and PF MPO concentrations may be a marker of the severity of the disease.

peer reviewed | Objective-To compare measurements of myeloperoxidase (MPO) in plasma, laminar tissues, and skin obtained from control horses and horses given black walnut heartwood extract (BWHE). Animals-22 healthy 5- to 15-year-old horses. Procedures-Horses were randomly assigned to 4 groups as follows: a control group given water (n = 5) and 3 experimental groups given BWHE (17) via nasogastric intubation Experimental groups consisted of 5, 6, and 6 horses that received BWHE and were euthanatized at 1.5, 3, and 12 hours after intubation, respectively. Control horses were euthanatized at 12 hours after intubation. Plasma samples were obtained hourly for all horses. Laminar tissue and skin from the middle region of the neck were harvested at the time of euthanasia. Plasma and tissue MPO concentrations were determined via an ELISA; tissue MPO activity was measured by use of specific immunologic extraction followed by enzymatic detection. Results-Tissues and plasma of horses receiving BWHE contained significantly higher concentrations of MPO beginning at hour 3. Laminar tissue and skin from horses in experimental groups contained significantly higher MPO activity than tissues from control horses. Concentrations and activities of MPO in skin and laminar tissues were similar over time. Conclusions and Clinical Relevance-in horses, BWHE administration causes increases in MPO concentration and activity in laminar tissue and skin and the time of increased MPO concentration correlates with emigration of WBCs from the vasculature. These findings support the hypothesis that activation of peripheral WBCs is an early step in the pathogenesis of acute laminitis.

peer reviewed | [en] OBJECTIVE: Treatment of orofacial tumors in dogs is associated with high morbidity and reliable prognostic factors are lacking. Dynamic contrast-enhanced computed tomography (DCECT) can be used to assess tumor perfusion. The objectives of this study were to describe the perfusion parameters of different types of orofacial tumors and to describe the changes in perfusion parameters during radiotherapy (RT) in a subset of them. ANIMALS: 11 dogs with orofacial tumors prospectively recruited. CLINICAL PRESENTATION AND PROCEDURES: All dogs had baseline DCECT to assess blood volume (BV), blood flow (BF), and transit time (TT). Five dogs had repeat DCECT during megavoltage RT. RESULTS: 5 squamous cell carcinomas, 3 sarcomas, 1 melanoma, 1 histiocytic sarcoma, and 1 acanthomatous ameloblastoma were included. Blood volume and BF were higher in squamous cell carcinomas than in sarcomas, although no statistical analysis was performed. At repeat DCECT, 4 dogs showed a reduction in the size of their tumor during RT. Among these dogs, 3 showed an increase in BV and BF and 1 a decrease in these parameters between the baseline and the follow-up DCECT. The only dog whose tumor increased in size between the first and the second DCECT showed a decrease in BV and BF. CLINICAL RELEVANCE: Perfusion parameters derived from DCECT were described in a series of dogs with various types of orofacial tumors. The results suggest that epithelial tumors could have higher BV and BF than mesenchymal tumors, although larger sample sizes are needed to support these preliminary findings.

peer reviewed | Abstract Title: Equine neutrophil elastase in plasma, laminar tissue, and skin of horses administered black walnut heartwood extract (BWHE) Author(s) Name(s) & Institutional Affiliation(s): Geoffroy de la Rebière de Pouyade1, 2, Laura. M. Riggs3, J. N. Moore4, Thierry Franck1, 2, Ginette Deby-Dupont2, David J. Hurley4,5, and Didier Serteyn1, 2 1-Department of Clinical Sciences, Large Animal Surgery, Faculty of Veterinary Medicine B41, University of Liège, Sart Tilman, 4000 Liège, Belgium 2-Center for Oxygen Research and Development, Institute of Chemistry B6a, University of Liège, Sart Tilman, 4000 Liège, Belgium 3-Department of Equine Surgery, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA 70803 4-Department of Large Animal Medicine, College of Veterinary Medicine, University of Georgia, Athens, GA 30602 5-Department of Population Health, College of Veterinary Medicine, University of Georgia, Athens, GA 30602 Take Home Message: Elastase concentrations are increased in the integument of horses during the developmental phase of BWHE-induced laminitis. Introduction: Current evidence suggests the involvement of neutrophils and myeloperoxidase (MPO), a strong oxidative enzyme in the development of laminitis. Elastase is a protease recognized as a component of inflammatory disorders. Neutrophil elastase concentrations were measured in plasma, skin and laminar tissues from control horses and horses given BWHE Materials & Methods (including Statistical Analyses): The study included 4 groups, a control group (water) and 3 experimental groups (BWHE). Plasma was obtained hourly until euthanasia of BWHE-treated horses at 1.5 (n=5), 3 (n=6), and 12 hours (n=10 plasma/n=6 tissue) and control horses at 12 hours (n=7 plasma/n=5 tissue). Elastase concentrations were determined via ELISA. Log-transformed data were analyzed with 1-way ANOVA (tissues) and unpaired t-test (plasma); significance was set at P<0.05. Results: Plasma elastase concentrations in the BWHE group were significantly higher at 6, 8 and 10h compared to the control group. Concentrations in skin and laminar tissue were significantly higher at 3 and 12h in the BWHE group. Elastase concentrations were significantly higher in skin than in lamina at 12h in BWHE group. Discussion: Increased elastase concentrations are consistent with dynamic changes in circulating leukocytes in horses given BWHE. Collectively, MPO and elastase have the potential to cause significant tissue damage during the development of laminitis. Clinical Relevance: Elastase may become an important target as new treatments for laminitis are developed. Conclusion: Elastase concentrations increase in the integumentary system during the development of laminitis and may participate in the disintegration of the hoof basal membrane.