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.
Frydendal C, Nielsen KB, Berg LC, van Galen G, Adler DMT, Andreassen SM, Jacobsen S.To investigate neutrophil gelatinase-associated lipocalin (NGAL) concentrations in serum and synovial fluid (SF) from horses with joint inflammation. Methods: Experimental studies and retrospective clinical study. Methods: Serum and SF samples were available from healthy horses (n = 19), clinical cases, and horses with experimental joint inflammation. Clinical cases included horses with (n = 10) or without (n = 10) septic arthritis. Experimental intra-articular inflammation was induced by lipopolysaccharide (LPS; n = 7, severe inflammation), lidocaine (n = 6, moderate inflammation), or mepivac...
Salami F, Ghodrati M, Parham A, Mehrzad J.Bone marrow mesenchymal stem cells (BM-MSCs), as multipotent cells with self-renewal and plastic-adherent properties, have immunomodulatory effects on immune cells, including neutrophils. These cells are in close proximity in bone marrow (BM) sinusoids with non-multiplicative immature neutrophils. BM-MSCs exert their immunomodulatory effects on adjacent cells both directly (cell-to-cell contact) and indirectly (secretion of soluble factors). The aim of this study was to evaluate the effect of equine bone marrow mesenchymal stem cells (BM-MSCs) on the expression of some pro- and anti-apoptotic ...
Amaral A, Fernandes C, Rebordão MR, Szóstek-Mioduchowska A, Lukasik K, Pinto-Bravo P, Telo da Gama L, Jan Skarzynski D, Ferreira-Dias G.Neutrophils can originate neutrophil extracellular traps (NETs). Myeloperoxidase (MPO) is a peroxidase found in NETs associated to equine endometrosis and can be inhibited by 4-aminobenzoic acid hydrazide (ABAH). Metallopeptidases (MMPs) participate in extracellular matrix stability and fibrosis development. The objectives of this in vitro work were to investigate, in explants of mare's endometrium, (i) the ABAH capacity to inhibit MPO-induced collagen type I (COL1) expression; and (ii) the action of MPO and ABAH on the expression and gelatinolytic activity of MMP-2/-9. Explants retrieved from...
Zhang M, Dickinson RM.Multiple red, raised nodules multifocally distributed along the serosal surface of the normal and the nonviable jejunum were identified in a 24-year-old neutered male horse undergoing surgery for removal of the strangulating lipoma around the jejunum. Histologically, these nodules consisted of many significantly and variably dilated, blood-filled vascular channels lined by a single layer of flattened, well-differentiated endothelial cells with occasional thrombi within a mildly thickened fibrous stroma. A diagnosis of intestinal angiomatosis was proposed. To the best of the authors' knowledge,...
Davis KU, Sheats MK.Asthma is a common and debilitating chronic airway disease that affects people and horses of all ages worldwide. While asthma in humans most commonly involves an excessive type 2 immune response and eosinophilic inflammation, neutrophils have also been recognized as key players in the pathophysiology of asthma, including in the severe asthma phenotype where neutrophilic inflammation predominates. Severe equine asthma syndrome (sEAS) features prominent neutrophilic inflammation and has been increasingly used as a naturally occurring animal model for the study of human neutrophilic asthma. This...
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...
Anderson SL, Duke-Novakovski T, Robinson AR, Townsend HGG, Singh B.The objective of this study was to determine the effect of pulmonary intravascular macrophage depletion on systemic inflammation and ex vivo neutrophil apoptosis using an experimental model of intestinal ischemia and reperfusion injury in horses. Neutrophils were isolated before and after surgery from horses that were randomized to three treatment groups, namely, sham celiotomy (CEL, = 4), intestinal ischemia and reperfusion (IR, = 6), and intestinal ischemia and reperfusion with gadolinium chloride treatment to deplete pulmonary intravascular macrophages (PIMs, IRGC, = 6). Neutrophil apopt...
Anderson MJ, Ibrahim AS, Cooper BR, Woolcock AD, Moore GE, Taylor SD.Sepsis is associated with ascorbic acid (AA) depletion and critical illness-related corticosteroid insufficiency (CIRCI) in humans. Objective: Intravenous infusion of lipopolysaccharide (LPS) would (a) decrease endogneous AA concentrations, (b) induce CIRCI and (c) administration of a combination of AA and hydrocortisone (HC) would have decreased indices of inflammation compared to either drug alone. Methods: Thirty-two healthy horses. Methods: Randomized placebo-controlled experimental trial. Horses were assigned to 1 of 4 groups (saline, AA and HC, AA only, or HC only). Treatments were admin...
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...
Miró J, Gutiérrez-Reinoso M, da Silva JA, Fernandes C, Rebordão MR, Alexandre-Pires G, Catalán J, Ferreira-Dias G.Collagen fibers and inflammatory cells are the basis for jenny endometrium Kenney and Doig's classification developed for the mare. The infiltration of a large number of eosinophils in the jenny endometrium is intriguing. Eosinophil and fibroblast produced IL33, which has been related to fibrosis development and chronicity. This work on the endometrium consisted of (i) quantification of collagen type I (COL1A2), type III (COL3A1), and IL33 transcripts; (ii) histological localization and quantification of COL1 and COL3 proteins; and (iii) eosinophil and neutrophil count and correlation with col...
Kutasi O, Fehér O, Sárdi S, Balogh N, Nagy A, Moravszki L, Bódai E, Szenci O.West Nile virus (WNV) is a zoonotic arbovirus transmitted by mosquitoes between wild birds (natural hosts) and other vertebrates. Horses and humans are incidental, dead-end hosts, but can develop severe neurological disorders. Owing to the close contact of cerebrospinal fluid (CSF) with the extracellular fluid of the brain, the analysis of CSF composition can reflect central nervous system (CNS) impairments enabling the diagnosis and understanding of various neurodegenerative CNS disorders. Our objective was to compare the findings from the CSF samples of horses with neuroinvasive WNV infectio...
Stucchi L, Alberti E, Stancari G, Conturba B, Zucca E, Ferrucci F.Mild-moderate equine asthma (MEA) is a common disease of young racehorses characterized by inflammation of the lower airways diagnosed by cytological examination of bronchoalveolar lavage fluid (BALF), and it is considered an important cause of poor performance. The most common parameter of athletic capacity associated with horse performance is the speed at 4 mmol/L of lactate (VLA4). The aim of the present work was to evaluate the relation between the different BALF inflammatory cells and VLA4 in racehorses affected by MEA. A population of Standardbred racehorses that underwent an accurate pr...
Sundman EA, Chigerwe M, Hunyadi LM.Exercise-induced pulmonary hemorrhage (EIPH) and asthma in barrel racing horses is a common disease across the United States. Limited information is available on non-infectious respiratory diseases in this population, the interaction between these two diseases, and the occurrence of both EIPH and asthma in the horse. The purpose of this study was to evaluate the bronchoalveolar lavage (BAL) fluid cytological results of barrel racing horses with EIPH, asthma, or both. A retrospective study was conducted using the medical records of horses that presented with cough and decreased athletic perform...
Albornoz A, Morales N, Uberti B, Henriquez C, Burgos RA, Alarcon P, Moran G.Neutrophils participate in innate immunity as the first line of host defence against microorganisms. However, persistent neutrophil activity and delayed apoptosis can be harmful to surrounding tissues; this problem occurs in diverse inflammatory diseases, including asthma-affected horses. Previous studies in horses with acute lung inflammation indicated that treatment with tamoxifen (TX), a selective oestrogen receptor modulator, produces a significant decrease in bronchoalveolar lavage fluid (BALF) neutrophil content. The aim of this study was to investigate the effect of tamoxifen and its me...
Lilliehöök I, Bröjer J, Nostell K, Kindahl H, Tvedten HW, Edner A, Hagman R.Endotoxemia is a common and severe disease of horses. Most previous studies have monitored changes caused by a bolus dose of endotoxin over short time periods. Objective: We aimed to describe inflammatory responses to endotoxin with inflammatory and hematologic markers monitored over a longer time than has been performed in the past using more prolonged endotoxin exposures. Methods: Escherichia coli O55:B5 endotoxin was administered as a 6-hour continuous intravenous infusion of lipopolysaccharide (LPS) to eight horses. Blood cell counts, and prostaglandin F -metabolite (PGM), serum amyloid A ...
Salinas C, Espinosa G, Morales N, Henríquez C, Morán G, Gajardo G, Uberti B.Obesity is a highly prevalent condition in horses. Dysfunctional neutrophil activity has been reported in metabolically healthy obese humans, but minimal data exist regarding horses. The present study evaluated the effect of obesity on apoptosis, phagocytosis and oxidative burst activity of peripheral blood neutrophils from lean and obese non-insulin dysregulated horses. Seven lean (BCS, body condition score 4-6/9) and five obese (BCS 8-9) horses were enrolled in the study. All animals underwent two metabolic tests (OGT, oral glucose test; IRT, insulin response test) before their selection to ...
Caissie MD, Gartley CJ, Scholtz EL, Hewson J, Johnson R, Chenier T.Persistent breeding-induced endometritis (PBIE) is a major cause of infertility in mares. Endometrial inflammation that persists until embryonic descent ultimately results in early embryonic death. A poor endometrial biopsy grade (IIb or III) has been identified as a risk factor for PBIE. Intrauterine fluid accumulation (>2 cm in depth), pathologic endometrial edema, and elevated intrauterine neutrophil levels are all clinical features of PBIE. Commonly applied treatment options include uterine lavage and oxytocin therapy. N-acetyl cysteine (NAC), a mucolytic used to treat bacterial endome...
Adler DMT, Serteyn D, Franck T, Jørgensen E, Christophersen MT, Denwood M, Verwilghen DR.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. 17 adult horses. 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 MC...
Amaral A, Fernandes C, Rebordão MR, Szóstek-Mioduchowska A, Lukasik K, Gawronska-Kozak B, Telo da Gama L, Skarzynski DJ, Ferreira-Dias G.Neutrophil extracellular traps (NETs) fight endometritis, and elastase (ELA), a protease found in NETs, might induce collagen type I (COL1) accumulation in equine endometrium. Metallopeptidases (MMPs) are involved in extracellular matrix balance. The aim was to evaluate the effects of ELA and sivelestat (selective elastase inhibitor) on MMP-2 and MMP-9 expression and gelatinolytic activity, as well as the potential inhibitory effect of sivelestat on ELA-induced COL1 in equine endometrium. Endometrial explants from follicular (FP) and mid-luteal (MLP) phases were treated for 24 or 48 h with ELA...
Brankston G, Greer AL, Marshall Q, Lang B, Moore K, Hodgins D, Hennessey JTG, Beeler-Marfisi J.Ambient pollution is associated with the development and exacerbation of human asthma, but whether air pollution exposure is associated with lower airway inflammation in horses has not been fully evaluated. The Air Quality Health Index (AQHI) is an online tool used by asthmatic Ontarians to modify their outdoor activity when ambient pollution is high. A single AQHI value, falling on a scale from 1 to 10, is calculated from measurements of fine particulate matter (PM), nitrogen dioxide (NO), and ozone (O). Increased AQHI values predict an increased risk for presenting to a health care provider ...
Hue E, Orard M, Toquet MP, Depecker M, Couroucé A, Pronost S, Paillot R, Richard EA.Few data on cytokine profiles in bronchoalveolar lavage fluid (BALF) are available for racehorses with mild/moderate equine asthma (EA); cytological diagnosis being most frequently made from only one lung. The purpose of the study was to compare cytokine mRNA expressions and protein concentrations in BALF from both lungs. As part of a larger study, 250 ml saline was randomly instilled in one lung and 500 ml in the contralateral lung of 30 clinically healthy Standardbred racehorses. This procedure was repeated 72 h later, inversing the volume per lung. Cytological cut-off values for diagnosis o...
Köhne M, Mönnig F, Papin J, Schöniger S, Tönissen A, Rohn K, Martinsson G, Schoon HA, Sieme H.The effects of standard uterine body and hysteroscopic insemination on endometrial health were investigated. For this purpose, 33 mares were assigned to five different protocols: control (no insemination; n = 7), sham AI (sham uterine body insemination; n = 6), sham HysAI (sham hysteroscopic insemination; n = 7), standard AI (standard uterine body insemination, 300 × 106 progressively motile sperms (PMS); n = 7) and HysAI (hysteroscopic insemination, 100 × 106 PMS; n = 6). Sampling included uterine swabbing for microbiological examination, cytology for determination of polymorphonucle...
Godbout M, Vargas A, Hélie P, Bullone M, Lavoie JP.To use a biopolymer delivery system to investigate the ability of interleukin (IL)-4 to recruit neutrophils into subcutaneous tissues of equids. Methods: 16 horses and 2 ponies. Methods: Animals were assigned to 3 experiments (6/experiment). Effects of recombinant equine (Req) IL-4 (100, 250, or 500 ng/site) versus a positive control (ReqIL-8; 100 ng, 250 ng, or 1 μg/site) and a negative control (Dulbecco PBSS or culture medium) on neutrophil chemotaxis were assessed after SC injection into the neck with an injectable biopolymer used as the vehicle. Tissue samples including the biopolymer plu...
Michalska J, Nowicka B, Wessely-Szponder J.We hypothesized that in horses with naturally occurring joint disorders, the neutrophil response, acute phase response (APR), and oxidative stress parameters elevated significantly and are markers of increased inflammatory response in these conditions. Therefore, the first aim of the study was to evaluate neutrophil response, oxidative status, and APR. The neutrophil activity was assessed on the basis of elastase, myeloperoxidase (MPO), and alkaline phosphatase release, whereas free radical generation was assessed on the basis of nitric oxide and superoxide production. Acute phase response was...
Skarzynski DJ, Szóstek-Mioduchowska AZ, Rebordão MR, Jalali BM, Piotrowska-Tomala KK, Leciejewska N, Łazarczyk M, Ferreira-Dias GM.The innate and adaptive immune mechanisms are key components of regulation of reproductive physiological function and uterine disorders in equine uterus. The predominant immunological response in equine endometrium, characterized by an innate immune response, occurs under estrogens influence, in the follicular phase. Although, the increase in immune-related genes in equine endometrium during estrus has been suggested to play a role in uterine clearance after mating, immune cells and their product, i.e. cytokines play also mandatory role in the luteal development and maintenance, regression of ...
Fernandez NJ, Gilroy CV, Wagg CR, Kwong GPS, Roy MF.The detection of band neutrophils and toxic change via microscopic blood smear review is vitally important, as their presence indicates systemic inflammation. However, in-clinic evaluation of WBC morphology is often limited. Objective: We aimed to determine the agreement between expert raters in the detection of bands and toxic change. Methods: Three board-certified clinical pathologists each evaluated 109 blood smears from horses with acute disease, and 19 control smears from healthy horses. The pathologists determined if bands were present, and if so, the percentage of bands present. They a...
Fingerhut L, Ohnesorge B, von Borstel M, Schumski A, Strutzberg-Minder K, Mörgelin M, Deeg CA, Haagsman HP, Beineke A, von Köckritz-Blickwede M....Equine recurrent uveitis (ERU) is considered one of the most important eye diseases in horses and typically appears with relapsing inflammatory episodes without systemic effects. Various disorders have been described as an initial trigger, including infections. Independent of the initiating cause, there are numerous indications that ERU is an immune-mediated disease. We investigated whether neutrophil extracellular traps (NETs) are part of the ERU pathogenesis. Therefore, vitreous body fluids (VBF), sera, and histological sections of the eye from ERU-diseased horses were analyzed for the prese...
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...
Espinosa G, Plaza A, Schenffeldt A, Alarcón P, Gajardo G, Uberti B, Morán G, Henríquez C.Polymorphonuclear neutrophils (PMN) are the largest leukocyte population in the blood of most mammals including horses, and play an important defensive role in many infectious diseases. However, the mechanisms that increase PMN as one of the main cellular subsets in the defense against pathogens could also be involved in the pathophysiology of dysregulated inflammatory conditions. Mesenchymal stem/stromal cells (MSCs) are a heterogeneous population with a modulatory potential on the inflammatory response and are known to interact with nearly all cells of the immune system, including PMN. In th...
Allen KJ, Tennant KV, Franklin SH.Published studies vary as to whether epithelial cells are included in differential counts for tracheal wash (TW) and bronchoalveolar lavage (BAL) cytology in horses. The aim of this study was to determine whether inclusion or exclusion of epithelial cells affects interpretation of airway cytology. Using criteria of >20% TW neutrophils, >10% BAL neutrophils and/or >5% BAL mast cells to indicate airway inflammation, there was a change in categorisation from 'normal' to 'abnormal' in 21%, 4% and 8% horses, respectively, when epithelial cells were excluded from differential counts. It is recommend...
Katavolos P, Ackerley CA, Clark ME, Bienzle D.Horses suffer from recurrent airway obstruction, an asthma-like condition induced by repeat inhalation of environmental substances present in barn air. Clara cell secretory protein (CCSP) is much reduced during active inflammation when neutrophils predominate in the airways, and in chronic asthmatics. We sought to investigate morphologic and functional interactions of CCSP with neutrophils. Bronchoalveolar and blood neutrophils from healthy control animals, and from animals with recurrent airway obstruction in remission and exacerbation, were evaluated by immuno-cytochemistry and immuno-electr...
Segabinazzi LGTM, Canisso IF, Podico G, Cunha LL, Novello G, Rosser MF, Loux SC, Lima FS, Alvarenga MA.Microorganisms, including pathogenic or opportunistic bacteria and fungi, may gain access to the uterus during breeding, and infectious endometritis plays a major role in equine subfertility. This study aimed to assess the post-breeding inflammatory response, endometrial culture, and embryo recovery of mares susceptible to persistent breeding-induced endometritis (PBIE) treated with plasma-rich (PRP) or -poor (PPP) plasma. Mares (n = 12) susceptible to PBIE had three cycles randomly assigned to receive intrauterine infusions of lactate ringer solution (LRS, control), or autologous PRP or PPP p...
Hurley DJ, Parks RJ, Reber AJ, Donovan DC, Okinaga T, Vandenplas ML, Peroni JF, Moore JN.Administration of black walnut heartwood extract (BWHE) via nasogastric tube induces acute laminitis in horses. However, the processes responsible for the development of laminitis, including laminitis induced with BWHE, remain unclear. The results of recent studies indicate that administration of BWHE initiates an inflammatory response in the laminar tissues and that this response may be due to extravasation of activated leukocytes from the circulation. This study examines the effects of BWHE administration on the dynamics of circulating neutrophils and monocytes, and the capacity of blood leu...
Tessier L, Côté O, Clark ME, Viel L, Diaz-Méndez A, Anders S, Bienzle D.Severe equine asthma is a chronic inflammatory condition of the lower airways similar to adult-onset asthma in humans. Exacerbations are characterized by bronchial and bronchiolar neutrophilic inflammation, mucus hypersecretion and airway constriction. In this study we analyzed the gene expression response of the bronchial epithelium within groups of asthmatic and non-asthmatic animals following exposure to a dusty hay challenge. After challenge we identified 2341 and 120 differentially expressed genes in asthmatic and non-asthmatic horses, respectively. Gene set enrichment analysis of changes...
Troedsson MH, Liu IK, Thurmond M.In vitro phagocytosis and chemotaxis of uterine and blood-derived polymorphonuclear neutrophils (PMNs) were compared in mares with different resistance to chronic uterine infection (CUI). Both the primary in vitro function of PMNs and the role of uterine environmental factors on PMN function were investigated. The uteri of mares susceptible to (n = 6) and resistant to CUI (n = 5) were inoculated with 5 x 10(6) Streptococcus zooepidemicus when the mares were in estrus. Uterine secretions in addition to uterine and blood-derived PMNs were sampled at 5 and 24 h later. During a subsequent estrus, ...
Baumber J, Vo A, Sabeur K, Ball BA.Contaminating leukocytes in the ejaculate are an important source of reactive oxygen species (ROS) in human semen. When present in sufficient numbers, they can have a detrimental influence on sperm function in humans. Unfortunately, there is little published information regarding the importance of leukocytes in stallion semen. The objectives of this study were to determine the production of hydrogen peroxide (H2O2) by activated equine neutrophils and to examine the effect of this ROS production on equine sperm motility in vitro. Motile equine spermatozoa (two ejaculates each from four stallion...
Millerick-May ML, Karmaus W, Derksen FJ, Berthold B, Robinson NE.Particle mapping within stalls has demonstrated a relationship between particulate concentrations and visible accumulations of tracheal mucus. However, measurement of breathing zone particulate concentrations, which is the most accurate way to measure exposure, has not been performed to determine the relationship between exposures and accumulations of tracheal mucus sufficient to affect performance. Objective: To compare breathing zone particulate exposures between horses with tracheal mucus scores (MS) previously demonstrated to affect performance (MS≥2) and those without (MS = 0) visible t...
Black SJ, Lunn DP, Yin C, Hwang M, Lenz SD, Belknap JK.The mechanisms that initiate the pathophysiologic changes in the digital laminae in equine laminitis are poorly understood. Due to the fact that (1) the horse at risk of laminitis has many similarities clinically to the human sepsis patient and (2) our recent finding of marked laminar proinflammatory cytokine expression at the developmental time point of the black walnut extract (BWE) model of laminitis, we tested the possibility that, similar to organ damage in human sepsis, leukocyte emigration is an early event in laminitis. Using immunoperoxidase methods with an anti-equine CD13 monoclonal...
Yoshikawa H, Yasu T, Ueki H, Oyamada T, Oishi H, Anzai T, Oikawa M, Yoshikawa T.To evaluate the possibility that Streptococcus equi subsp. zooepidemicus (S.z) the causative bacterial agent of equine shipping fever pneumonia (ESFP), as well as to investigate its pathogenesis, 10 horses (seven Thoroughbreds and three Anglo-Arab species, ranging from 2-4 years in age) were experimentally inoculated, via an endoscope, into bronchus of the lung lobe with a dose of 30 ml of 1-7 x 10(8) CFU/ml of S.z. After inoculation, autopsy and pathological examinations were sequentially conducted 30 min, 1, 2, 3, 4, 17, 20 hr and 2 weeks later. Pneumonia induced by the intrapulmonary inocul...
Kang H, Bienzle D, Lee GKC, Piché É, Viel L, Odemuyiwa SO, Beeler-Marfisi J.Severe equine asthma (SEA) is a common, debilitating lower airway inflammatory disorder of older horses. Alveolar macrophages (AMs) survey inhaled particulates from barn sources causing them to switch from an anti-inflammatory to a proinflammatory phenotype, resulting in neutrophil recruitment to the lung. This proinflammatory switch may contribute to the development and prolongation of SEA. Validated antibodies to identify the cells involved in the pathogenesis of SEA are lacking. In this study, monoclonal antibodies against CD90, CD163, and CD206 were tested for reactivity with equine leukoc...
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 ...
Borlone C, Morales N, Henriquez C, Folch H, Olave C, Sarmiento J, Uberti B, Moran G.Neutrophils participate in innate immunity as the first line of host defense against microorganisms. However, exacerbated neutrophil activity can be harmful to surrounding tissues; this is important in a range of diseases, including allergic asthma and chronic obstructive pulmonary disease in humans, and equine asthma (also known as recurrent airway obstruction (RAO). Tamoxifen (TX) is a non-steroidal estrogen receptor modulator with effects on cell growth and survival. Previous preliminary studies showed that TX treatment in horses with induced acute pulmonary inflammation promoted early apop...
Barussi FC, Bastos FZ, Leite LM, Fragoso FY, Senegaglia AC, Brofman PR, Nishiyama A, Pimpão CT, Michelotto PV.This research evaluated the effects of bone marrow-derived mononuclear cells (BMMCs) on the inflammatory process in the equine recurrent airway obstruction (RAO). Eight horses in RAO clinical score were divided into cell therapy group (Gcel) treated with a single intratracheal dose of BMMCs, and dexamethasone group (Gdex) treated with 21days of oral dexamethasone. The horses were clinically revaluated on days 7 and 21, together with cytological evaluation of the BALF, and detection of inflammatory markers (interleukins [IL]-10, -4, and -17, and interferon γ and α). There were decreases in re...
Aktan I, Dunkel B, Cunningham FM.Activated platelets can contribute to host defense through release of products with bactericidal actions such as antimicrobial peptides and reactive oxygen species (ROS), as well as by forming heterotypic aggregates with neutrophils and enhancing their antimicrobial properties. Whilst release of vasoactive mediators from equine platelets in response to stimuli including bacterial lipopolysaccharide (LPS) has been documented, neither ROS production, nor the effects of activated platelets on equine neutrophil ROS production, have been reported. This study first sought evidence that activated equ...
Aharonson-Raz K, Lohmann KL, Townsend HG, Marques F, Singh B.Heaves, an obstructive neutrophilic airway inflammation of horses, is triggered by dust components such as endotoxin and has similarities to human asthma. Pulmonary intravascular macrophages (PIMs) increase horses' sensitivity to endotoxin-induced lung inflammation; however, their role in an airborne pathology remains unknown. Therefore, we investigated the role of PIMs in the development of heaves in horses. Clinical and inflammatory responses were evaluated following induction of heaves by moldy hay exposure and PIM depletion with gadolinium chloride (GC). Mares (N = 9) were exposed to four ...
Jacobsen S, Berg LC, Tvermose E, Laurberg MB, van Galen G.Neutrophil gelatinase-associated lipocalin (NGAL) has been shown to be a useful marker of kidney injury in people and dogs, but has not been described in horses. Objective: The aim of the study was to validate a commercially available porcine-specific ELISA to measure serum concentrations of equine NGAL. Methods: Intra- and interassay imprecisions were evaluated by multiple measurements on equine serum pools. Assay inaccuracy was determined by the linearity under dilution. Overlapping performance was assessed by measuring NGAL concentrations in horses with normal and elevated serum creatinine ...
Wilmink JM, Veenman JN, van den Boom R, Rutten VP, Niewold TA, Broekhuisen-Davies JM, Lees R, Armstrong S, van Weeren PR, Barneveld A.Wound healing proceeds faster in ponies than in horses and complications during healing, such as wound infection, occur less frequently in ponies. Earlier studies suggested that this difference might be related to differences in the initial post traumatic inflammatory response. Objective: That polymorphonuclear leucocyte (PMN) function and profiles of humoral factors in local inflammatory processes are different in horses and ponies. Methods: PMNs were isolated from venous blood of horses and ponies. Chemotaxis and reactive oxygen species (ROS) production was determined. Tissue cages were impl...
Boschwitz JS, Timoney JF.The antiphagocytic property of equine fibrinogen for Streptococcus equi subsp. equi strain CF32 was examined in vitro. The results of bactericidal assays demonstrated that the presence of fibrinogen enhanced the ability of overnight and early log-phase cultures of strain CF32 to resist killing by equine neutrophils by 12-fold and seven-fold, respectively (p > 0.01). In addition, fibrinogen-coated bacteria treated with fibrinogen specific F(ab')2 fragments were 32% more susceptible to killing by equine neutrophils after opsonization in serum (p > 0.05), indicating that specific epitopes o...
Tiwari R, Qin A, Artiushin S, Timoney JF.Evasion of phagocytosis is an important virulence determinant of Streptococcus equi (S. equi subsp. equi), the cause of equine strangles and distinguishes it from the closely related but much less virulent S. zooepidemicus (S. equi subsp. zooepidemicus). We describe Se18.9, a novel H factor binding protein secreted by S. equi but not by S. zooepidemicus that reduces deposition of C3 on the bacterial surface and significantly reduces the bactericidal activity of equine neutrophils suspended in normal serum for both S. equi and S. zooepidemicus. Se18.9 is secreted abundantly by actively dividing...
Miskovic M, Couëtil LL, Thompson CA.The effects of long-term environmental management on airway obstruction and inflammation in horses with recurrent airway obstruction (RAO) are unknown. Objective: Horses with RAO maintained in low-dust environments have persistent airway obstruction and neutrophilic inflammation. Methods: Study horses were treated for RAO and then maintained in low-dust environments with no medical management. Horses were classified into 3 groups by years after diagnosis: 1 year (time 1, n = 9), 2-3 years (time 2, n = 7), and 5-6 years (time 3, n = 8). The comparison groups were age-matched healthy horses. Met...
Dawson DR, Nydam DV, Price CT, Graham JE, Cynamon MH, Divers TJ, Felippe MJ.To investigate the effect of opsonization of Rhodococcus equi with R. equi-specific antibodies in plasma on bacterial viability and phagocyte activation in a cell culture model of infection. Methods: Neutrophils and monocyte-derived macrophages from 6 healthy 1-week-old foals and 1 adult horse. Methods: Foal and adult horse phagocytes were incubated with either opsonized or nonopsonized bacteria. Opsonization was achieved by use of plasma containing high or low concentrations of R. equi-specific antibodies. Phagocyte oxidative burst activity was measured by use of flow cytometry, and macrophag...
Mönki J, Saastamoinen M, Karikoski N, Rajamäki M, Raekallio M, Junnila J, Särkijärvi S, Norring M, Valros A, Oranen Ben Fatma S, Mykkänen A.Bedding materials affect stable air hygiene, and thus the development and exacerbation of equine asthma. There is limited knowledge concerning the effects of different types of bedding material on equine lower airway inflammation. The objective of our study was to investigate the effects of bedding materials on respiratory signs, tracheal mucus score, and lower airway cytology in healthy adult horses. The study design was a prospective controlled study, and the subjects were healthy adult riding school horses (n = 32) from a single stable. Wood shavings were compared to peat, which was used as...
Liu M, Liu T, Bordin A, Nerren J, Cohen N.Toll-like receptor 9 (TLR9) activation stimulates protective immune responses against intracellular pathogens by phagocytes, including neutrophils. This study examined TLR9-mediated neutrophil activation in neonatal foals. Unmethylated CpGs, ligands for TLR9, were used to stimulate equine neutrophils, either purified or in contact with other peripheral blood leukocytes. Rhodococcus equi was used as another stimulus in parallel. TLR9 mRNA was constitutively expressed at a similar level in purified equine neutrophils across different ages from birth to adulthood, and expression was not affected ...
Moore RM, Bertone AL, Bailey MQ, Muir WW, Beard WL.Histomorphologic/morphometric evaluation, leukocyte scintigraphy, and myeloperoxidase activity were used to determine whether neutrophils accumulate in the large colon of horses during low-flow ischemia and reperfusion. Twenty-four adult horses were assigned to 1 of 3 groups: group 1, sham-operated (n = 6); group 2, 6 hours of ischemia (n = 9); and group 3, 3 hours of ischemia and 3 hours of reperfusion (n = 9). Low-flow ischemia of the large colon was induced in horses of groups 2 and 3 by reducing colonic arterial blood flow to 20% of baseline. Radiolabeled (99mTc) autogenous neutrophils wer...
Yánez-Ortiz I, Catalán J, Mateo-Otero Y, Dordas-Perpinyà M, Gacem S, Yeste N, Bassols A, Yeste M, Miró J.Jenny shows a large endometrial reaction after semen influx to the uterus with a large amount of polymorphonuclear neutrophils (PMN) migrating into the uterine lumen. PMN act as a sperm selection mechanism through phagocytosis and NETosis (DNA extrudes and, together with proteins, trap spermatozoa). While a reduced percentage of spermatozoa are phagocytosed by PMN, most are found to be attached to neutrophil extracellular traps (NETs). This selection process together with sperm metabolism produces a large amount of reactive oxygen species (ROS) that influence the reproductive success. The pres...
Henry MM, Moore JN.Increasing evidence has demonstrated the importance of monocyte procoagulant activity (PCA) in the pathogenesis of coagulopathies in a variety of diseases. Because endotoxin precipitated coagulopathies are common sequelae to intestinal ischemia/endotoxemia in the equine species, we investigated the ability of equine peripheral blood monocytes to express PCA. Monocytes isolated from five healthy adult horses were incubated in vitro with Escherichia coli endotoxin (10 micrograms), and the PCA was measured by the ability of cellular lysates to accelerate the clotting times of equine plasma in a m...
Franck T, Kohnen S, Grulke S, Neven P, Goutman Y, Peters F, Pirotte B, Deby-Dupont G, Serteyn D.In the horse, the inflammation response to various pathologies (intestinal strangulations, laminitis, etc.) involves an excessive stimulation of the polymorphonuclear neutrophils releasing reactive oxygen species (ROS) and myeloperoxidase (MPO). The aim of the present work was to study the effect of natural polyphenols, curcuminoids and tetrahydrocurcuminoids (THC) on isolated stimulated equine neutrophils and on the activity of purified MPO. The ROS production and the release of MPO by activated neutrophils were measured by chemiluminescence and ELISA techniques, respectively. The activity of...
Dewachi O, Joubert P, Hamid Q, Lavoie JP.Heaves, a condition associated with airway neutrophilia, is believed to result from an allergic response to environmental dust particles. However, the contribution of neutrophils to the allergic response is poorly understood. It has been hypothesized that Th2-type cytokines can directly activate neutrophils to produce pro-inflammatory mediators. The present study focused on the presence of receptors for the Th2-type cytokines interleukin (IL)-5 and IL-9 on peripheral blood neutrophils of horses with heaves. Neutrophils were isolated from peripheral blood of horses with heaves (n=7), and normal...
Cardwell JM, Wood JL, Smith KC, Newton JR.Rates of airway inflammation in young racehorses decrease with time but it is not clear whether this is associated with increasing age or time exposed to the training environment. The structure of the British National Hunt (NH) population allowed closer examination of this relationship. Objective: To compare rates of inflammatory airway disease diagnosed by tracheal sampling (trIAD), and its components, in NH racehorses by age and training history and with published rates in young racehorses. Methods: A prospective, longitudinal study was conducted on 5 NH yards over 2 years. Period sample pre...
Martin EM, Till RL, Sheats MK, Jones SL.In many equine inflammatory disease states, neutrophil activities, such as adhesion, migration, and reactive oxygen species (ROS) production become dysregulated. Dysregulated neutrophil activation causes tissue damage in horses with asthma, colitis, laminitis, and gastric glandular disease. Non-steroidal anti-inflammatory drugs do not adequately inhibit neutrophil inflammatory functions and can lead to dangerous adverse effects. Therefore, novel therapies that target mechanisms of neutrophil-mediated tissue damage are needed. One potential neutrophil-targeting therapeutic is the PGE1 analog, m...
