Neutrophils are a type of white blood cell that play a significant role in the immune system of horses. They are involved in the body's defense mechanisms against infections and are typically one of the first responders to microbial invasion or tissue damage. Neutrophils function by engulfing and destroying pathogens through processes such as phagocytosis and the release of antimicrobial substances. In horses, the number and activity of neutrophils can be indicative of immune status and are often assessed in the context of infectious diseases, inflammation, and other health conditions. This page compiles peer-reviewed research studies and scholarly articles that explore the function, regulation, and clinical implications of neutrophils in equine health.
Weiss DJ, Evanson OA, Fagliari JJ, Valberg S.To determine whether platelets become activated and form platelet-neutrophil aggregates during near-maximal treadmill exercise in horses. Methods: 4 Thoroughbreds. Methods: Horses were subjected to 4 standardized exercise tests on a treadmill, and blood samples were collected before exercise, at treadmill speed of 12 m/s, and 5 minutes after exercise. Flow cytometric techniques were used to identify activated platelets, and flow cytometric and microscopic techniques were used to identify platelet-neutrophil aggregates. Results: Platelet-neutrophil aggregates increased from 2.8 +/- 0.4% at rest...
Brazil TJ, Rossi AG, Haslett C, McGorum B, Dixon PM, Chilvers ER.The synthetic formylpeptide fMLP is widely used as a model chemoattractant and secretagogue for mammalian neutrophils. Despite possessing fMLP receptors, equine neutrophils do not produce superoxide anions in response to fMLP and there is no inflammatory reaction in the horse when fMLP is injected intradermally. The functional capability of these receptors was investigated after pretreatment with recognized priming agents. Purified neutrophils were pretreated with lipopolysaccharide (LPS), platelet-activating factor (PAF), or tumor necrosis factor alpha (TNF-alpha) and superoxide anion generat...
Foster AP, Cunningham FM.Equine eosinophils and neutrophils are believed to play an important part in the protection of horses against parasitic and bacterial invasion. Eosinophils may also play a key role in the pathogenesis of equine inflammatory conditions such as the allergic skin disease, insect hypersensitivity. The factors which stimulate the respiratory burst of equine eosinophils and neutrophils are poorly understood. The first aim of the present study was to determine the effects of the phorbol ester, phorbol myristate acetate (PMA), which is believed to activate intracellular protein kinase C, and opsonised...
Weiss DJ, Evanson OA, McClenahan D, Fagliari JJ, Jenkins K.To determine whether platelets are hyperaggregable or form platelet-neutrophil aggregates during the prodromal stages of acute laminitis of ponies. Methods: Healthy adult ponies: 8 experimental and 6 control. Methods: Acute laminitis was induced by oral administration of corn starch and wood flour to 8 ponies, and indices of platelet activation were evaluated. Blood samples were collected before and at 4, 8, 12, 24, 28, and 32 hours after carbohydrate administration, and PCV, total plasma protein concentration, platelet count, activated clotting time, whole blood recalcification time, spontane...
Toews AR, Campbell JR.To assess the value of CBC as a preoperative test in healthy horses undergoing cryptorchidectomy. Methods: Retrospective study. Methods: 117 horses. Methods: Medical records were reviewed to identify horses that had had cryptorchidectomy. Of the 117 horses identified, 102 were found that did not have a known medical condition believed to adversely affect surgery. Preoperative CBC was assessed in terms of abnormalities detected and effects of these abnormalities on development of complications during and after surgery and patient management. Results: Of 102 CBC performed, 55 contained abnormali...
Cunningham FM, Andrews M, Landoni MF, Lees P.Calcium ionophore A23187 induced time and concentration dependent production of immunoreactive leukotriene (LT) B4 by equine heparinized whole blood in vitro. Time dependent production of immunoreactive LTB4 by equine neutrophils and immunoreactive LTC4 by equine eosinophils in vitro was also demonstrated. The 5-lipoxygenase activating protein (FLAP) inhibitors, BAY X 1005 and BAY Y 1015, produced concentration dependent inhibition of ionophore-induced LTB4 synthesis by equine whole blood (mean +/- SEM IC50s n = 5; 6.14 +/- 0.28 microM vs. 12.30 +/- 0.75 microM for BAY Y 1015 and BAY X 1005, r...
Gröndahl G, Johannisson A, Jensen-Waern M.The ability of equine plasma from different donors to enhance phagocytic capacity was assessed in neutrophils obtained from seven foals, aged 7-8 days (Study A), and from seven adult horses (Study B). Neutrophils were allowed to phagocytize fluorescent yeast cells opsonized with plasma from one of three donors or with pooled serum, all previously frozen (-18 degrees C) and thawed. The results were analysed by flow cytometry. In study A, fresh autologous foal serum was also used for opsonization, and in study B, heat-inactivated plasma and pooled serum were used in addition to untreated samples...
Raidal SL, Bailey GD, Love DN.To evaluate the effect of transportation on lower respiratory tract contamination and peripheral blood neutrophil function in horses and to compare results from transported horses with those obtained in earlier experiments from horses confined with heads elevated. Methods: A prospective study. Methods: Six horses were transported by road for 12 h. Clinical and haematological examination, transtracheal aspiration and cell function studies were conducted before and after transportation. Results obtained after transportation were compared to pre-transportation values. Results: After transportatio...
Moore T, Wilcke J, Chilcoat C, Eyre P, Crisman M.Equine neutrophils (PMN) play a critical role in inflammatory processes in horses. The objective of this study was to characterize equine PMN function ex vivo following stimulation with calcium ionophore A23187 (A23187) and phorbol myristate acetate (PMA). These stimulants trigger different branches of the PMN activation process that occurs in vivo. Equine PMN were isolated from the whole blood of six clinically normal geldings using a one-step discontinuous Percoll gradient technique. Neutrophil aggregation, degranulation, and superoxide anion production were evaluated in assay systems which ...
Furr M, Tinker MK, Edens L.This study was conducted to develop an equation for the prediction of outcome in neonatal foals undergoing treatment in an intensive care unit (ICU). Fifty-three physical examination, historical, and clinicopathologic variables were analyzed from the records of 99 neonatal foals (< 14 days of age) treated in the neonatal ICU of the Equine Medical Center. The outcome was recorded and the results were categorized into either surviving or nonsurviving groups. The mean values for the 2 groups were compared, and variables that differed significantly between the two groups were retained and used ...
Marr KA, Foster AP, Lees P, Cunningham FM, Page CP.The effect of antigen challenge on the state of activation of peripheral blood neutrophils from horses with chronic obstructive pulmonary disease (COPD) has been determined by measuring neutrophil superoxide anion formation. Prior to a seven-hour antigen challenge superoxide anion production by neutrophils from asymptomatic horses with COPD and normal horses in response to platelet activating factor (PAF) (with and without cytochalasin B), serum treated zymosan (STZ) and phorbol myristate acetate (PMA) was similar. Agonist-induced superoxide production by neutrophils from symptomatic COPD and ...
Koivunen AL, Maisi P, Konttinen YT, Sandholm M.The gelatinolytic activity in tracheal aspirates (TA) of horses with chronic obstructive pulmonary disease (COPD) was analyzed using SDS-PAGE-gelatin-gel electrophoresis (zymography) and compared to TAs from healthy controls. The 110-90 kD MMP-9 type gelatinase was high in symptomatic disease phases (permanent disease 0.46 +/- 0.15, p < 0.001; or intermittent disease 0.47 +/- 0.12, p < 0.001) compared to healthy controls (0.10 +/- 0.07). Similarly, the overall gelatinolytic activity, the activity in high-mw gelatinolytic bands (210-190 and 150 kD) and in proteolytically processed fragments in ...
Reilas T, Katila T, Mäkelä O, Huhtinen M, Koskinen E.Intrauterine fluid (IUF) was collected using a tampon from mid-oestrous mares (n = 57) with and without ultrasonically detectable accumulations of free intraluminal fluid. Bacteria were cultured and neutrophils counted from all samples (n = 57). Total protein concentration, trypsin-inhibitor capacity (TIC), and plasmin, beta-glucuronidase (B-Gase) and N-acetyl-beta-D-glucosaminidase (NAGase) activities were determined in 27 IUF samples. The motility of spermatozoa in the presence of IUF, IUF extended with Kenney's medium (1:1) and Kenney's medium alone was analysed in 9 samples using a Hamilto...
Koivunen AL, Maisi P, Konttinen YT, Prikk K, Sandholm M.The collagenolytic activity and its sensitivity to doxycycline inhibition in tracheal aspirates (TA) of horses with chronic obstructive pulmonary disease (COPD) was analyzed with SDS-PA gel electrophoresis (SDS-PAGE), using Type 1 collagen as the substrate. Both autoactive and total collagenase activities were significantly higher in TAs of horses with symptomatic COPD than in TAs of healthy horses. Doxycycline inhibition studies suggest that most of the TA collagenase is of the neutrophil type (MMP-8), but some is derived from other cells such as fibroblasts and monocyte/macrophages (MMP-1) a...
Lakritz J, Wilson WD, Watson JL, Hyde DM, Mihalyi J, Plopper CG.To determine whether oral administration of erythromycin alters the inflammatory response to bronchoalveolar lavage (BAL) in young horses. Methods: 12 healthy, unweaned, mixed-breed foals of either sex, between 2 and 4 months old. Methods: BAL was performed; 250 ml of phosphate-buffered saline solution (300 mOsm, pH 7.4) was administered in 50-ml aliquots. Foals were carefully monitored for 4 days, then erythromycin base (25 mg/kg of body weight, PO, q 12 h) was given to foals of the treated group. After 4 days, foals were reanesthetized, and the same lung was relavaged. Cytologic examination ...
MacDonald MH, Benton HP.To determine the influence of activated equine neutrophils on sulfated glycosaminoglycan metabolism of equine articular cartilage in vitro. Methods: Articular cartilage explants harvested from the metacarpophalangeal joints of 7 horses. Methods: Proteoglycan degradation and synthesis were measured by release of glycosaminoglycan from the explants, and incorporation of [35S]sulfate into newly synthesized glycosaminoglycan. Results: Activated equine neutrophils significantly increased the release of glycosaminoglycan from explant matrix and the magnitude of that response was influenced by durati...
Hiemstra PS, Maassen RJ, Stolk J, Heinzel-Wieland R, Steffens GJ, Dijkman JH.Antileukoprotease (ALP), or secretory leukocyte proteinase inhibitor, is an endogenous inhibitor of serine proteinases that is present in various external secretions. ALP, one of the major inhibitors of serine proteinases present in the human lung, is a potent reversible inhibitor of elastase and, to a lesser extent, of cathepsin G. In equine neutrophils, an antimicrobial polypeptide that has some of the characteristics of ALP has been identified (M. A. Couto, S. S. L. Harwig, J. S. Cullor, J. P. Hughes, and R. I. Lehrer, Infect. Immun. 60:5042-5047, 1992). This report, together with the catio...
Foreyt WJ, Lagerquist JE.Peripheral blood neutrophils from horses, cattle, and Rocky Mountain bighorn sheep (Ovis canadensis canadensis) were evaluated for susceptibility to cytotoxin-dependent lysis of different biotypes and serotypes of Pasteurella haemolytica of domestic sheep, cattle, bighorn sheep, or mountain goat (Oreamnos americana) origin utilizing a cytotoxicity assay which measures the degree of bacteria cytotoxin-killing of neutrophils. All isolates of P. haemolytica (biotypes A and T) were noncytotoxic to horse neutrophils. Thirteen of 18 R haemolytica biotype A isolates were cytotoxic (> 50% neutrophi...
Fairbairn SM, Marr KA, Lees P, Cunningham FM, Page CP.Antigen challenge is known to cause the recruitment of neutrophils to the lungs of horses with chronic obstructive pulmonary disease (COPD). To evaluate a possible role of platelet activating factor (PAF) in this process, the effects of PAF on the distribution of radiolabelled neutrophils were compared in normal horses and asymptomatic horses with COPD. Changes in lung function, heart rate and the distribution of platelets and eosinophils were also measured. PAF (5 ng kg-1 intravenously) caused immediate but transient increases in the number of radiolabelled neutrophils in the lungs and a conc...
Muñoz MC, Doreste F, Ferrer O, González J, Montoya JA.A 16-year-old, male, Hanoverian horse had a three-month history of weight loss, hirsutism and polyuria/polydypsia. Examinations revealed neutrophilia, lymphopenia, hyper glycaemia and abnormalities in hepatic function. A tentative diagnosis of hyperadrenocorticism was made. The results of thyroid-releasing hormone and combined dexamethasone suppression and ACTH stimulation tests suggested the presence of a pituitary adenoma. The horse was treated with pergolide and beneficial clinical and biochemical responses were observed within one to six months.
Marr KA, Fairbairn SM, Page CP, Lees P, Cunningham FM.Antigen challenge involving exposure to straw and mouldy hay for 7 h produced lung function changes and neutrophil recruitment to the lungs in horses with chronic obstructive pulmonary disease (COPD). During the challenge, an increase in radiolabelled neutrophils in the lungs occurred, together with increased respiratory rate and pleural pressure. The role of platelet activating factor (PAF) in antigen-induced neutrophil accumulation, and increased pleural pressure and respiratory rate was investigated by administering the PAF receptor antagonist WEB 2086 to asymptomatic COPD horses prior to a...
Raulo SM, Hyyppa S, Räsänen LA, Pösö AR.The activities of lysosomal enzymes, such as beta-glucuronidase and N-acetyl-beta-D-glucosaminidase, have been shown to increase in muscle after endurance exercise. We examined whether measurable activities of lysosomal enzymes are present in equine plasma and whether the exercise-induced changes in the muscle are reflected in plasma. Six trained Standardbred trotters performed three exercise bouts with 1 h intervals and the same procedure was repeated 3 days later. Venous blood samples and muscle biopsies from the middle gluteal muscle were taken before and after exercise. The activities of b...
Moore BR, Krakowka S, Cummins JM, Robertson JT.Natural human interferon-alpha (nHuIFN alpha) was administered to actively training Standardbred racehorses with inflammatory airway disease (IAD). Inflammatory airway disease was characterized by poor exercise performance and inflammation and exudate in the upper and lower airway. Placebo, 50, 150, or 450 units(U) of nHuIFN alpha was administered orally for 5 consecutive days to eight horses per treatment group in a double-blind, randomized block design. Response to nHuIFN alpha was monitored by semiquantitative endoscopic examination score and cytologic examination of bronchoalveolar lavage ...
Tvedten HW, Korcal D.The differential leukocyte counts performed by an automated hematology analyzer, the Technicon H-1E Hematology System, and traditional microscopic method (M-Diff) from blood samples of 129 horses, 40 cattle, and 140 cats were compared. The comparison was repeated after selected subsets of data were created by deleting samples with certain patterns suggesting error with the automated differential cell count (A-Diff). The two methods had good comparison of results for neutrophils and lymphocytes in all three species. Results for equine monocytes correlated moderately well between the two methods...
Olszewski MA, Robinson NE, Yu MF, Derksen FJ.During acute bouts of recurrent airway obstruction (heaves) in horses, neutrophils that are capable of increased production of reactive oxygen species accumulate in the airways. In the study reported here, the effect of hydrogen peroxide (H2O2; 1 microM to 0.1M), one of these reactive oxygen species products, on the responses of isolated trachealis muscle of horses was determined. Before and after incubation with H2O2, contractile responses to acetylcholine, electrical field stimulation (EFS), 127 mM KCl, and relaxation responses to isoproterenol and activation of the nonadrenergic noncholiner...
Dixon PM, Railton DI, McGorum BC.The majority of 270 adult horses with chronic pulmonary diseases had excessive volumes of respiratory secretions (RS) present in their trachea, elevated tracheal RS neutrophil ratios and lowered arterial oxygen partial pressures. Some control horses, had inexplicably elevated trachael RS neutrophil ratios. Only the chronic obstructive pulmonary disease (COPD) affected group had significantly elevated bronchoalveolar lavage fluid (BALF) neutrophil ratios. Maximal intrapleural pressure changes (max dPpl) were raised in 48% of COPD cases and rarely with other pulmonary disorders. Arterial pH or c...
Foster AP, Lees P, Cunningham FM.Platelet activating factor (PAF) mimics the effects of Culicoides antigen by inducing oedema and inflammatory cell accumulation in the dermis of horses with the allergic skin disease, sweet itch. PAF could therefore contribute to antigen-induced inflammatory changes in these horses. We now report that intravenous administration of the PAF receptor antagonist WEB 2086 (3 mg kg-1), at a dose that inhibited the vascular and cellular responses to PAF in sweet itch horses, reduced Culicoides antigen-induced oedema at 1 h by 73% and at 8 h by 71% (p < 0.05). Neutrophil accumulation and eosinophil...
McKane SA, Rose RJ, Evans DL.Twenty-four Thoroughbred and twelve Standardbred racehorses aged between 2 and 6 years, presented for reported poor racing performance, underwent clinical exercise testing. During the last 10 s of exercise at each speed throughout an incremental speed exercise test on a treadmill inclined at a 10% slope, samples of arterial blood and expired gases were collected. Maximum oxygen uptake and the partial pressures of oxygen and carbon dioxide in arterial blood were determined. These values were compared between the two breeds of horses and also with reference to cytological findings of bronchoalve...
Riber C, Rubio MD, Marquez F, Pinedo M, Muñoz A, Castejon F.Clinical blood chemistry was studied on 13 Andalusian horses with laminitis. Ten healthy horses were used as controls. Blood samples were taken within the first 24 hr after the onset of the disease in Obel grade 3 before any type of treatment was given. No significant differences were observed in the number of red blood cells and white blood cells between Andalusian horses affected by laminitis and healthy animals. Significant decrease in the number of neutrophils and increased number of lymphocytes were noted. A significant increase in the enzymatic activity of CPK, LDH and AST in these anima...
Kohn CW, Swardson C, Provost P, Gilbert RO, Couto CG.Myeloid and megakaryocytic bone marrow hypoplasia in association with moderate to profound neutropenia was observed in 8 young Standardbred horses sired by the same stallion; 7 horses were intermittently thrombocytopenic. Evaluation of serial neutrophil counts in 2 horses suggested that a cyclic variation in neutrophil numbers was present, that lymphocyte numbers increased when neutrophil counts decreased, and that platelet counts decreased when neutrophil counts decreased. Preliminary bone marrow cultures indicated that myeloid progenitor cells were present and that these cells were able to r...
Flisińska-Bojanowska A, Gill J, Grzelkowska K.In the blood of 11 foals and their lactating mothers (Standardbred) diurnal changes in the cortisol level, neutrophil number and lysozyme activity were studied during the first 13 weeks of life. The investigations began when a foal reached 7 days of age and were repeated every two weeks till 13 weeks of age. Blood samples were taken from the jugular vein every 4 hours for one day. Experiments were repeated in two following years. In the first year 6 mares and 6 foals born by these mares were examined, and in the second year--5 of the mares from the first year and the 5 new foals borne by them....
Watson ED.Uterine washings were found to promote neutrophil mediated killing of Streptococcus zooepidemicus. Depletion of complement and/or specific antibody from the washings significantly reduced bactericidal activity. Phagocytosis of yeast by uterine washings was complement dependent. Inhibition of the classical pathway significantly reduced opsonic activity indicating that, in addition to direct activation via the alternate pathway, antibody may also be involved in yeast phagocytosis.
Seahorn TL, Gaunt SD, Berry C.Hematologic and rheologic variables were examined in a group of 13 horses with intestinal colic and a control group of 6 horses. All horses had been recently transported to the veterinary teaching hospital, and blood samples were obtained during initial examination. There were no significant differences in blood neutrophil count or plasma fibrinogen concentration between the groups, and PCV was significantly increased in horses with intestinal colic. Cell filterability was measured by passing uniform concentrations of blood, erythrocytes, and neutrophils through micropore filters. There were n...
Wyse CA, Nixon C, McLaughlin M, Dowell FJ, Philbey AW.The aim of this study was to characterise the expression of haem oxygenase-1 (HO-1) in healthy lung tissue from horses and to measure its activity. Samples of lung tissue were collected from six horses euthanased for reasons other than respiratory disease. HO-1 expression and activity were detected in type II alveolar epithelial cells, macrophages and neutrophils in all the samples examined. The activity was dependent on the presence of NADPH and inhibited quantitatively by the addition of increasing concentrations of a competitive inhibitor of HO-1, tin mesoporphyrin IX.
Blanchard TL, Orsini JA, Garcia MC, Elmore RG, Youngquist RS, Bierschwal CJ.A retrospective study was done on total white blood cell (WBC) and blood neutrophil counts of 41 mares referred to one of two veterinary hospitals for correction of dystocia. The mares were 2 to 19 years of age and included draft, light, and pony breeds. The WBC and neutrophil counts were performed at varying intervals from time of admission to 10 d after delivery of the feti. Retrospective analyses of WBC and neutrophil counts from 10 normal foaling mares from two Pennsylvania breeding farms (Thoroughbred and Trakehner) and from 14 normal foaling pony mares were done as controls. Mean WBC (10...
Nerren JR, Payne S, Halbert ND, Martens RJ, Cohen ND.Rhodococcus equi is an intracellular pathogen of macrophages that causes rhodococcal pneumonia in foals and immunocompromised people. Evidence exists that neutrophils play a vital role in resistance to infection with R. equi; however, the means by which neutrophils exert their effects have not been clearly defined. In addition to directly killing bacteria, neutrophils also may exert a protective effect by linking innate and adaptive immune responses. In the present study we evaluated the cytokine expression profiles of adult equine neutrophils in response to stimulation with isogenic strains o...
Pycock JF, Allen WE.Streptococcal endometritis was induced experimentally in pony mares during oestrus. Uterine fluid was collected 30, 60, 120 or 240 minutes later and tested for its effect on the in vitro morphology and chemotaxis of equine neutrophils by two independent methods. The maximal response occurred between 30 and 60 minutes after infection and persisted until 240 minutes. The chemo-attractant contained both heat labile and heat stable components and the latter appeared to be active at low concentrations.
Heyneman RA, Vercauteren RE.The subcellular components of purified neutrophil leukocytes from horse blood were fractionated by isopyknic equilibration in sucrose and metrizamide gradients. Five classes of particles have been identified: dense azurophil granules containing the bulk of the lysosomal acid hydrolase and peroxidase activity (A); less dense particles, containing all the lysozyme activity, but not resolved from a second population of azurophils B, and particles of low density, biochemically characterized as a plasma membrane fraction (C). Isopyknic equilibration in sucrose disclosed a minor membrane fraction (D...
Perez-Ecija A, Buzon-Cuevas A, Aguilera-Aguilera R, Gonzalez-De Cara CA, Mendoza FJ.Preanalytical factors such as storage time and temperature are proved to induce marked artifactual changes in hematological parameters in horses, small animals and humans. These errors can mirror findings typical of endotoxemia, leading to dangerous misdiagnosis. Since donkeys are common in warm climates and remote regions, blood samples from this species can be subjected to long lasting travels from the farm to the nearest laboratory, frequently under suboptimal conditions. Moreover, as other equids, donkeys are prone to suffer endotoxemia. Nonetheless, stability has not been evaluated in sam...
Wauters J, Franck T, Pille F, Martens A, Demeyere K, Sys S, Serteyn D, Gasthuys F, Meyer E.Myeloperoxidase (MPO) is a protein of interest due to its involvement in equine pathologies. Until now, results in equine diagnostic research were achieved through extracellular MPO detection. However, studying the cellular MPO content in neutrophils has revealed important insights in human diseases. This study aimed to develop a technique for the specific detection of MPO on the single cell level defining a flow cytometric protocol for the detection of both equine surface-bound and cellular MPO. Both indirect and direct labeling techniques are described which include the comparison of two sec...
Siemsen DW, Kirpotina LN, Malachowa N, Schepetkin IA, Porter AR, Lei B, DeLeo FR, Quinn MT.The development of new advances in understanding the role of neutrophils in inflammation requires effective procedures for isolating and purifying neutrophils. Methods for isolating human neutrophils are fairly standard, and some are covered in other chapters of this volume and previous editions. However, procedures for isolating neutrophils from nonhuman species used to model human diseases vary from those used in isolating human neutrophils and are not as well developed. Since neutrophils are highly reactive and sensitive to small perturbations, the methods of isolation are important to avo...
Heyneman RA, Bruyninckx WJ, Vercauteren RE.Two distinct groups of acid phosphatase containing granules were characterized in neutrophils, each group displaying different multiple forms of the enzyme. The heavy granule acid phosphatase showed a lysosomal location. A second lighter group of particles contained a thermolabile, thiol-dependent acid p-nitrophenyl and alpha-naphtylphosphatase, an enzyme clearly different from lysosomal acid phosphatase. Acid phosphatase activity from eosinophil leukocytes appeared to be totally associated with the typical eosinophil granules. On mechanical disruption of these particles, an acid phosphatase w...
Pycock JF, Allen WE, Porter DJ, Boyd EH.Two independent assay systems were used to study the effect of three antibacterial preparations on in vitro morphology and chemotaxis of equine neutrophils. Incubation of neutrophils with high (200 micrograms/ml) and medium (20 micrograms/ml) concentrations of neomycin impaired their response to standard chemoattractants. Trimethoprim/sulfadoxine (0.4/2.0 micrograms/ml-40/200 micrograms/ml) and benzylpenicillin (0.25-25 micrograms/ml) had no effect. Neutrophils collected from geldings 2 and 24 h after neomycin (5 mg/kg) administration had impaired responses to standard chemoattractants. Benzyl...
Wauters J, Martens A, Pille F, Dumoulin M, Gasthuys F, Sys S, Meyer E.Synovial fluid samples from culture-confirmed infected joints (n=13), joints with pronounced non-infectious synovitis (n=11) and healthy joints (n=14) were collected from 24 equine patients and seven slaughterhouse horses. The samples from the joints with non-infectious synovitis and healthy joints served as negative controls. After isolation, counting and identification of neutrophils, the percentage viability, and the proportion apoptotic and necrotic neutrophils were determined by flow cytometry. Viability was significantly higher in infected samples compared to the controls. A significant ...
Coignoul FL, Bertram TA, Roth JA, Cheville NF.Neutrophils from 4 pony foals, 3 lactating pony mares, and 3 nonlactating mares were evaluated ultrastructurally and by in vitro function tests. Neutrophils from foals had significantly (P = 0.05) less random migration than neutrophils from mares; values in tests for iodination and Staphylococcus aureus ingestion were also lower with foal neutrophils. Neutrophils from lactating mares had lower responses to iodination, antibody-dependent cell-mediated cytotoxicity, and random migration tests than did neutrophils from nonlactating mares. Ultrastructurally, granule concentration did not differ si...
Weiss DJ, Geor RJ, Burris SM, Smith CM.Pentoxifylline has been reported to improve peripheral vascular circulation by altering the flow properties of blood. To determine if the hemorrheological effects of pentoxifylline were mediated by alterations in neutrophil function and/or flow properties, we evaluated the drug's effects on equine neutrophils in vitro. Pentoxifylline, at a concentration of 1 x 10(-1) M, but not at concentrations of 1 x 10(-6) M to 1 x 10(-2) M, markedly suppressed neutrophil superoxide production, zymosan phagocytosis and adherence to nylon wool. Pentoxifylline failed to improve neutrophil filterability throug...
Fadok VA.Abstract- Urticarial eruptions, with or without pruritus, are common lesions in horses. The pathogenesis of these lesions can include immunological and other mechanisms. Research in the human field suggests that the mast cell co-ordinates the urticarial response by releasing a complex array of inflammatory mediators. Other cells, including the neutrophil, the eosinophil and the macrophage, may also play a role in the development of wheals. Elucidation of the role of many of these cells and mediators in the evolution of urticaria is only just beginning. Successful treatment of this dermatologic...
Asbury AC, Schultz KT, Klesius PH, Foster GW, Washburn SM.Ten mares, 5 resistant and 5 susceptible to bacterial endometritis, were examined for differences pertaining to the efficiency of phagocytosis of bacteria by neutrophils in the uterus. An assay for chemiluminescence was used to evaluate various schemes of opsonization and relate them to phagocytic rate. 123I-labelled albumin was used to measure protein migration to the inoculated uterus. Neutrophil numbers moving to the uterine lumen were determined. Before inoculation resistant mares were found to have substances in their uterine secretions that opsonized bacteria, resulting in effective phag...
Watson ED, Stokes CR, Bourne FJ.The function of blood and uterine luminal neutrophils from ovariectomized mares treated with ovarian steroids was investigated 18 h after intrauterine infusion of 1 X 10(9) Streptococcus zooepidemicus. Random migration of blood neutrophils under agarose was reduced by treatment with progesterone compared with that of neutrophils from oestradiol-treated and control mares. In-vitro addition of progesterone to blood neutrophils from acyclic ponies also reduced migration. Uterine neutrophils did not migrate under agarose which was probably an effect of bacterial phagocytosis. Hormone treatment had...
Zerbe H, Engelke F, Klug E, Schoon HA, Leibold W.An endometritis model was used to investigate the influence of degenerative endometrial changes (endometrosis) on functional parameters of uterine neutrophils in the horse. Six hours after intrauterine application of recombinant human interleukin-8 (rhIL-8), the uteri of 15 mares were flushed with phosphate-buffered saline. Quantitative and qualitative flow cytometric assays were then made to determine the absolute numbers, viability, phenotype, generation of reactive oxygen species (ROS), and phagocytic activity of immigrated polymorphonuclear neutrophilic granulocytes (PMN). Recombinant hIL-...
von Fellenberg R.The importance of proteases and protease inhibitors for the pathogenesis of pulmonary emphysema and chronic bronchitis of the horse is described. Endogenous elastases from neutrophil granulocytes and macrophages, which probably provoke emphysema in the human being, are not relevant in horse emphysema. Exogenous elastases from different species of streptomyces may be responsible for emphysema generation in this species. Part of the exogenous elastases are poorly or not inhibited at all by the equine blood protease inhibitors especially by alpha 1-protease inhibitors. A disorder similar to genet...
Woodward MC, Andrews FM, Kearney MT, Del Piero F, Hammerberg B, Pucheu-Haston CM.To characterize the response of skin of nonallergic horses following ID injection of polyclonal rabbit anti-canine IgE (anti-IgE) and rabbit IgG. Methods: 6 healthy horses. Methods: Skin in the cervical area was injected ID with anti-IgE and IgG. Wheal measurements and skin biopsy specimens were obtained before and 20 minutes and 6, 24, and 48 hours after injection. Tissue sections were evaluated for inflammatory cells at 4 dermal depths. Immunohistochemical analysis for CD3, CD4, and CD8 was performed, and cell counts were evaluated. Results: Anti-IgE wheals were significantly larger than IgG...
Kohn CW, Swardson C, Provost P, Gilbert RO, Couto CG.Myeloid and megakaryocytic bone marrow hypoplasia in association with moderate to profound neutropenia was observed in 8 young Standardbred horses sired by the same stallion; 7 horses were intermittently thrombocytopenic. Evaluation of serial neutrophil counts in 2 horses suggested that a cyclic variation in neutrophil numbers was present, that lymphocyte numbers increased when neutrophil counts decreased, and that platelet counts decreased when neutrophil counts decreased. Preliminary bone marrow cultures indicated that myeloid progenitor cells were present and that these cells were able to r...
Montgomery JB, Hamblin B, Suri SS, Johnson LE, New D, Johnston J, Kelly J, Wilson DG, Singh B.Ischemia followed by reperfusion leads to release of toxic molecules into the circulation, and these molecules may cause injury in remote organs such as the lung. Horses commonly suffer from episodes of intestinal ischemia-reperfusion (IR) due to intestinal twisting/strangulation followed by repair. Because there is no evidence of lung injury associated with IR in horses, we designed a study to characterize the intestinal IR-associated lung inflammation and determine the effect of lidocaine on lung inflammation in IR horses. Lung tissues were collected from non-anesthetized (n=4) and anestheti...
Conrado FO, Iapoce N, Batista-Linhares M, Lopez S, Matthews MH, McKinney CA, Rothacker C.An 18-year-old, grey, Thoroughbred Cross gelding was referred to the Cummings School of Veterinary Medicine at Tufts University following a 3-week history of low-grade fever of unknown origin, distal limb swelling, and weight loss. Clinical examination identified a few black, round, smooth nodules along the ventral aspect of the proximal tail. Transabdominal ultrasound showed a markedly enlarged heterogenous spleen, hyperechoic liver nodules, and evidence of peritonitis with fibrin deposition. A mature neutrophilia was noted on complete blood count with variable numbers of phagocytized granule...
Rutledge JJ, Paegelow J, Ritchey J, Singh A, Rizzi T, Murray C, Gilliam L, Crisman E, Williams NJ, Holbrook TC.The objective of this study was to determine the extent that nebulized glycosylated caffeic acid phenylether ester-4-O-alpha-D-glucopyranoside (G-CAPE) attenuates particulate-induced airway inflammation in healthy horses. Our hypothesis was that nebulization with G-CAPE would result in improved respiratory scores, higher arterial oxygen partial pressure, and less inflammatory airway infiltrates in horses with induced airway inflammation, compared with untreated controls. Five healthy adult horses were housed inside a climate controlled, closed barn on straw bedding and fed moldy grass hay for...
Zerbe H, Castilho LF, Engelke F, Mattos RC, Schuberth HJ, Klug E, Leibold W.Sufficient numbers of functionally competent polymorphonuclear neutrophil granulocytes (PMN) seem to be of major importance during the course of equine endometritis. In this study, we wanted to establish a method for cryopreservation of functionally competent neutrophils for an intended local endometritis therapy in mares. The separation of leucocytes by hypotonic lysis of whole blood from clinically healthy mares was superior to the separation by dextrose sedimentation. After suspension of the cells in the cryoprotective solution [equine plasma with 5% (v/v) dimethyl sulphoxide (DMSO)], the l...