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.
Birckhead EM, Das S, Tidd N, Raidal SL, Raidal SR.Septic synovitis and peritonitis are routinely diagnosed in horses based on clinical examination findings and laboratory assessment of synoviocentesis and abdominocentesis samples, respectively. Diagnosis is difficult in some cases because of an overlap in laboratory results for septic and non-septic inflammation. Neutrophil extracellular trap (NET) formation is part of the innate immune response against pathogens. Identifying and quantifying NETs, which have not been explored in clinical samples from horses with septic synovitis and peritonitis, to our knowledge, may be helpful in detecting i...
Morini M, Gobbo F, Rinnovati R, Romagnoli N, Peli A, Massarenti C, Spadari A, Pietra M.Equine asthma is a common respiratory disease that may affect horses of any age. The diagnosis of severe equine asthma (SEA) (historically referred as recurrent airway obstruction or RAO) is based mainly on the history of the animal and clinical signs, which are further supported by the cytological examination of the bronchoalveolar lavage (BAL). This can also be helpful in monitoring the inflammation of the lower airways in response to environmental management and medication. The cytocentrifugated preparation is usually considered the method of choice for BAL cytological interpretation. The a...
Westermann CM, de Bie AG, Olave C, de Grauw JC, Teske E, Couetil LL.Equine bronchoalveolar lavage (BAL) is usually performed with 250-500 mL of isotonic saline at pH 5.5. The acidic pH of saline may cause an increase in airway neutrophil count 48 h after BAL. Other isotonic solutions such as Ringer's solution, phosphate-buffered saline (PBS) or Plasma-Lyte 148 have a neutral pH of 7.4 and might be a better choice for BAL by not provoking inflammation and the influx of neutrophils into airways. BAL was performed in four healthy horses in four different lung lobes using four different solutions in a randomized crossover design. In each lobe, BAL was performed tw...
The aim of our study was to analyze circadian rhythm of the hematological profile of horses housed in a loose box and paddock during the different seasons (spring, summer, autumn, and winter). Blood samples were performed every 4 h for 48 consecutive hours. Red blood cells (RBCs), hemoglobin (HGB), hematocrit (HCT), white blood cells (WBCs), platelets (PLTs), and leukocyte subpopulations (neutrophils, basophils, eosinophils, lymphocytes, and monocytes) were analyzed, and, at the same time, environmental conditions were recorded. A statistically significant effect of housing conditions ( < 0...
Laurberg M, Saegerman C, Jacobsen S, Berg LC, Laursen SH, Hoeberg E, Sånge EA, van Galen G.Equine neonatal sepsis can be challenging to diagnose and prognosticate. Neutrophil gelatinase-associated lipocalin (NGAL), a new marker of renal damage and inflammation, can potentially be helpful. To evaluate NGAL in neonatal foals with sepsis, and assess its relation to outcome. Foals ≤ 14 days, with admission blood analysis and stored serum. NGAL was measured on stored serum from 91 foals. Foals were scored for sepsis and survival and categorized according to sepsis status (septic, sick non-septic, healthy, and uncertain sepsis status) and outcome groups (survivors and non-survivors). Th...
Hansen S, Otten ND, Spang-Hanssen L, Bendorff C, Jacobsen S.Studies in people have found neutrophil gelatinase-associated lipocalin (NGAL) concentrations are increased in asthma and can be used to distinguish between asthma subtypes. NGAL has not yet been investigated in equine asthma (EA). Objective: To investigate the ability of NGAL concentrations in bronchoalveolar lavage (BAL) fluid and serum to distinguish between control horses, horses with mild-moderate EA (MEA) and horses with severe EA (SEA). Methods: Retrospective cross-sectional study. Methods: Details of endoscopic examination including tracheal mucus score (TMS, scale 0-5) and BAL cytolog...
Amaral A, Cebola N, Szóstek-Mioduchowska A, Rebordão MR, Kordowitzki P, Skarzynski D, Ferreira-Dias G.Myeloperoxidase is an enzyme released by neutrophils when neutrophil extracellular traps (NETs) are formed. Besides myeloperoxidase activity against pathogens, it was also linked to many diseases, including inflammatory and fibrotic ones. Endometrosis is a fibrotic disease of the mare endometrium, with a large impact on their fertility, where myeloperoxidase was shown to induce fibrosis. Noscapine is an alkaloid with a low toxicity, that has been studied as an anti-cancer drug, and more recently as an anti-fibrotic molecule. This work aims to evaluate noscapine inhibition of collagen type 1 (C...
Oliver A, Conrado FO, Nolen-Walston R.Equine granulocytic anaplasmosis is a clinically significant and common disease of equids that has a broader prevalence than was once thought. The most common clinical signs include high fever and edema, with mild to mderate thrombocytopenia and lymphopenia typically noted on complete blood count. Subclinical cases are reported and many are self-limiting. Rare clinical presentations include neurologic disease, vasculitis, dysphagia, rhabdomyolysis, or bicavitary effusion. Most cases resolve rapidly with appropriate antimicrobial intervention.
Townsend M, Fowler B, Aulakh GK, Singh B.Endotoxin-induced diseases cause significant mortality and morbidity in the horse, leading to enormous economic damage to the equine industry. Neutrophils play a critical role in initiating the immune response in the lung. Pattern recognition receptors (PRRs) are programmed to recognize microbial structures unique to pathogens and mount an immune response. Pentraxin 3 (PTX3) is a PRR that is produced at sites of inflammation by many cell types upon stimulation by pro-inflammatory cytokines and agonists, such as endotoxins [also known as lipopolysaccharides (LPS)]. Pentraxin 3 recognizes and bi...
Salinas C, Barriga K, Albornoz A, Alarcon P, Quiroga J, Uberti B, Sarmiento J, Henriquez C, Ehrenfeld P, Burgos RA, Moran G.Neutrophils display an array of biological functions including the formation of neutrophil extracellular traps (NETs), web-like structures specialized in trapping, neutralizing, killing and preventing microbial dissemination within the host. However, NETs contribute to a number of inflammatory pathologies, including severe equine asthma. Tamoxifen (TX) is a selective estrogen receptor modulator which belongs to the triphenylethyllenes group of molecules, and which is used as a treatment in all stages of estrogen-positive human breast cancer. Our previous results suggest that tamoxifen can modu...
Ayo JO, Ake AS, Abimbola AA.The study aimed to evaluate effects of packing (load-carrying) and trekking on hematological and biochemical parameters in donkeys during the hot-dry season. Ten pack donkeys, aged 2-3 years, weighing 93 ± 2.7 kg were enrolled in the study and randomly divided into two groups of five animals each. Group 1 donkeys were subjected to packing + trekking while group 2 trekked only. All the donkeys trekked 20 km thrice, 1-day apart. Blood was collected before and after packing or trekking for hematological evaluation; and serum analyzed for electrolyte concentrations, and aspartate aminotransfera...
Jacobsen S, Mortensen CD, Høj EA, Vinther AM, Berg LC, Adler DMT, Verwilghen D, van Galen G.Neutrophil gelatinase-associated lipocalin (NGAL) has been suggested to be a highly sensitive and specific marker of joint infection in humans. The aim of the study was to investigate NGAL concentrations in synovial fluid (SF) from horses with septic synovitis, horses without septic synovitis, and horses with uncertain status. NGAL was measured in 177 admission samples obtained from 152 horses. From a subset of horses ( = 35), additional samples obtained sequentially over the course of treatment were available. Concentrations of NGAL were significantly higher in septic synovitis ( = 47 samples...
Olave CJ, Ivester KM, Couetil LL, Burgess J, Park JH, Mukhopadhyay A.Racehorses commonly develop evidence of mild asthma in response to dust exposure. Diets deficient in omega-3 polyunsaturated fatty acids (Ω-3) might exacerbate this response. Objective: To compare dust exposure, bronchoalveolar lavage fluid (BALF) cytology, and plasma Ω-3 and specialized pro-resolving mediators (SPM) concentrations amongst racehorses fed dry hay, steamed hay, and haylage. Methods: Forty-three Thoroughbred racehorses. Methods: Prospective clinical trial. Horses were randomly assigned to be fed dry hay, steamed hay, or haylage for 6 weeks. Measures of exposure to dust in the...
Regnier A, Landrevie C, Robin MC, Einsweiler D, Douet JY, Raymond-Letron I.A 6-year-old miniature Shetland pony mare was referred for evaluation of a left corneal mass, which developed from the healing tissue of a corneal traumatic ulceration that had occurred 4 weeks previously. On gross examination, a spherical, smooth-surfaced, and pink-colored lesion of about 1 cm in diameter was protruding from the left palpebral fissure. Ophthalmic examination revealed that it was attached to the scar tissue of the cornea, and that one corpora nigra was adherent to the posterior face of corneal wounded area, without sign of uveitis. The remainder of the ophthalmic examinatio...
Mild-moderate equine asthma (MEA) is a common respiratory disorder among racehorses, characterized by lower airway inflammation. Although it is generally agreed that MEA causes poor performance, contrasting results have been reported about the effects of tracheal mucus and the leukocyte populations of the bronchoalveolar lavage (BAL) on performance. This study aims to investigate the associations between airway inflammation and fitness parameters measured during an incremental treadmill test on Standardbred racehorses. For this purpose, the clinical records of 116 Standardbreds subjected to a ...
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...
Mainguy-Seers S, Beaudry F, Fernandez-Prada C, Martin JG, Lavoie JP.Extracellular vesicles (EVs) contribute to intercellular communication through the transfer of their rich cargo to recipient cells. The EVs produced by LPS-stimulated neutrophils from healthy humans and horses increase airway smooth muscle (ASM) proliferation, but the roles of neutrophil EVs in asthma are largely unexplored. The aim of this study was to determine whether neutrophil-derived EVs isolated during the remission or exacerbation of asthma influence ASM proliferation differentially. Peripheral blood neutrophils were collected during remission and exacerbation in eight horses affected ...
Cascardo B, Bernardes C, de Souza GN, Silva KM, Pires NR, de Alencar NX, Lessa DAB.Lower airway cytology has been proposed as a complementary diagnostic method to confirm the presence and quantifying the severity of exercise-induced pulmonary hemorrhage (EIPH). Bronchoalveolar lavage is usually preferred over tracheal wash (TW), yet the need for sedation imposes as a limitation for active sport horses in addition to be a highly invasive technique. This study aims to evaluate the diagnostic value of TW with Total Hemosiderin Score (THS) for monitoring EIPH in active Thoroughbred racehorses. A sampling of 47 randomly selected Thoroughbreds were endoscopically examined for EIPH...
Sage SE, Nicholson P, Peters LM, Leeb T, Jagannathan V, Gerber V.The transcriptomic profile of a cell population can now be studied at the cellular level using single-cell mRNA sequencing (scRNA-seq). This novel technique provides the unprecedented opportunity to explore the cellular composition of the bronchoalveolar lavage fluid (BALF) of the horse, a species for which cell type markers are poorly described. Here, scRNA-seq technology was applied to cryopreserved equine BALF cells. Analysis of 4,631 cells isolated from three asthmatic horses in remission identified 16 cell clusters belonging to six major cell types: monocytes/macrophages, T cells, B/plasm...
Dias De Castro LL, Oliveira Júnior GA, Perez BC, Carvalho ME, De Souza Ramos EA, Ferraz JBS, Molento MB.Cyathostomins are considered one of the most important parasites of horses. A group of horses within a herd can be responsible for eliminating the majority of parasite eggs. This phenotype might be explained by genetic factors. This study aimed to identify genomic regions associated with fecal egg count (FEC) and hematological parameters by performing a genomic-wide association study (GWAS) in Thoroughbred horses naturally infected with cyathostomins. Packed cell volume (PCV), differential leukocyte, and FEC were determined from 90 horses. All animals were genotyped using the Illumina Equine 7...
Boger BL, Manfredi JM, Yob C, Weber PSD, Jacobs CC.Diagnosis of equine septic arthritis is not straightforward, and increasing time between onset, diagnosis, and treatment can have serious consequences for quality of life. Defensins are used in diagnosis of human joint infection. The presence of beta defensins (BDs) in equine synovial fluid and their utility as a biomarker of sepsis has not been investigated; therefore, our objectives were to (1) compare in vitro gene expression of lipopolysaccharide (LPS)-stimulated equine neutrophils to unstimulated neutrophils and (2) compare BD protein expression from normal, aseptically inflamed, and sept...
Janssen P, Tosi I, Hego A, Maréchal P, Marichal T, Radermecker C.Asthma encompasses a spectrum of heterogenous immune-mediated respiratory disorders sharing a similar clinical pattern characterized by cough, wheeze and exercise intolerance. In horses, equine asthma can be subdivided into severe or moderate asthma according to clinical symptoms and the extent of airway neutrophilic inflammation. While severe asthmatic horses are characterized by an elevated neutrophilic inflammation of the lower airways, cough, dyspnea at rest and high mucus secretion, horses with moderate asthma show a milder neutrophilic inflammation, exhibit intolerance to exercise but no...
Protection against infectious diseases can be obtained with vaccines generating immunogenic response through a combination of humoral and cellular immunity. In this study haematological and serum protein electrophoretic profiles of horses vaccinated against herpesvirus 1 (EHV-1) and 4 (EHV-4) were evaluated. Blood samples were collected from 16 horses before (T0), after 24h, 48h, 72h, 1st week, 2nd week and 3rd week (T1I, T2I, T3I, T7I, T14I and T21I) from the first EHV vaccine-dose administration as well as before (TPRE), and after 24h, 48h, 72h, 1st week, 2nd week, 3rd week and 4th week (T1I...
Winther MF, Haugaard SL, Pihl TH, Jacobsen S.Neutrophil gelatinase-associated lipocalin (NGAL) is produced in response to inflammation in horses, but it has not yet been investigated as a biomarker in horses with the abdominal disease. Objective: To investigate NGAL in serum and peritoneal fluid in horses with the abdominal disease. Methods: Retrospective clinical study. Methods: Data from medical records of 270 horses admitted with acute abdominal disease (simple obstructions [n = 43], strangulating obstructions [n = 104], inflammatory abdominal disease [n = 99], non-strangulating intestinal infarction [NSII, n = 24]) and 9 heal...
Bayless RL, Sheats MK, Jones SL.Neutrophils play a major role in many equine conditions, including equine asthma, laminitis, and intestinal ischemia and reperfusion injury, and therefore represent an attractive target for innovative therapeutic approaches. Novel strategies for reducing neutrophilic inflammation include modulation of neutrophil functions and lifespan. Withaferin A (WFA) is a phytochemical with well-established and anti-inflammatory properties, but its direct effects on neutrophils are largely unknown. We hypothesized that WFA would inhibit adhesion, migration, and respiratory burst by equine neutrophils and...
Zandoná Meleiro MC, de Carvalho HJC, Ribeiro RR, da Silva MD, Salles Gomes CM, Miglino MA, de Santis Prada IL.Racehorses are constantly exposed to stress. Aiming to verify the state of blood components and cortisol alterations during their routine and after races, phagocytosis and oxidative neutrophil burst assays, serum cortisol determination, erythrocytes apoptosis evaluation, lymphoproliferation assays, and blood count tests were performed in thirty Thoroughbred racehorses, which were divided in two groups. The samples were taken right after races (moment 0 d), during rest periods (-11 d, +1 d, +3 d), and after training (-8, +2, +5). In both groups, the phagocytosis showed a decrease in percentage ...
Journal of proteomicsMarch 26, 2022
Volume 261 104571 doi: 10.1016/j.jprot.2022.104571
Koziy RV, Bracamonte JL, Yoshimura S, Chumala P, Simko E, Katselis GS.Septic arthritis (SA) is a life-threatening condition in horses, and identifying eradication of infection in equine SA is challenging. This study explored the discovery of putative biomarkers for the eradication of joint infection in horses. We performed proteomics analysis of synovial fluid (SF) and plasma from horses with experimental SA, non-septic lipopolysaccharide-induced arthritis, and controls. The point of eradication of infection in horses with SA was determined previously. We compared spectral intensities between groups as well as before and after the eradication of infection. Twent...
Lee GKC, Kang H, Beeler-Marfisi J, Sears W, Lillie BN, Bienzle D.Salivary scavenger and agglutinin (SALSA) is a secreted protein with various immunomodulatory roles. In humans, the protein agglutinates and inactivates microorganisms, and inhibits the release of pro-inflammatory cytokines. Saliva, which is rich in SALSA, accelerates bacterial phagocytosis, but SALSA's contribution is unclear. In horses, the functions of SALSA in inflammation remain undetermined, so they were investigated through phagocytosis and cytokine assays. Equine SALSA was purified from duodenal tissue, which contains abundant SALSA. To assess phagocytosis, fluorescently-labelled bacte...
Storms N, Medina Torres C, Franck T, Sole Guitart A, de la Rebière G, Serteyn D.Laminitis is a pathology of the equine digit leading to a failure of the dermo-epidermal interface. Neutrophil activation is recognized as a major factor in SIRS-associated laminitis. Less is known about the role of neutrophil activation in laminitis associated with metabolic disorders. The aim of this descriptive study was to observe whether myeloperoxidase is increased in the laminae during early stage laminitis in three horses subjected to a prolonged euglycemic hyperinsulinemic clamp (pEHC). After 48 h of pEHC-treatment, horses were subjected to euthanasia. Two healthy horses are used as c...
Krumrych W, Danek J, Markiewicz H, Gołyński M.The aim of this study was to analyze the chemiluminescence (CL) of peripheral blood in clinically healthy horses of different sexes and ages. The tests were carried out on 119 half- -breed horses, representing various forms of use (66 recreational horses and 53 sport horses). The test material was peripheral blood, which was collected under resting conditions, i.e. before physical activity related to the use of these animals. In the blood samples, spontaneous and stimulated CL with zymosan and phorbol myristate acetate were determined. It has been found that regular training effort increases t...
Robinson NE, Berney C, Eberhart S, deFeijter-Rupp HL, Jefcoat AM, Cornelisse CJ, Gerber VM, Derksen FJ.To investigate relationships between cough frequency and mucus accumulation, airway obstruction, and airway inflammation and to determine effects of dexamethasone on coughing and mucus score. Methods: 13 horses with recurrent airway obstruction (RAO) and 6 control horses. Methods: 6 RAO-affected and 6 control horses were stabled for 3 days. Coughing was counted for 4 hours before and on each day horses were stabled. Before and on day 3 of stabling, tracheal mucus accumulation was scored, airway obstruction was assessed via maximal change in pleural pressure (deltaPpl(max)), and airway inflamma...
Scocchi M, Bontempo D, Boscolo S, Tomasinsig L, Giulotto E, Zanetti M.Cathelicidins are precursors of defense peptides of the innate immunity and are widespread in mammals. Their structure comprises a conserved prepropiece and an antimicrobial domain that is structurally varied both intra- and inter-species. We investigated the complexity of the cathelicidin family in horse by a reverse transcription-PCR-based cloning strategy of myeloid mRNA and by Southern and Western analyses. Three novel cathelicidin sequences were deduced from bone marrow mRNA and designated equine cathelicidins eCATH-1, eCATH-2 and eCATH-3. Putative antimicrobial domains of 26, 27 and 40 r...
Bullone M, Joubert P, Gagné A, Lavoie JP, Hélie P.The severe form of equine asthma is associated with pathological changes of the peripheral airways and pulmonary parenchyma that are only partly described. Also, the relationship between these structural alterations and the percentage of neutrophils found within the airway lumen, assessed by bronchoalveolar lavage fluid (BALF) cytology, remains ill-defined. Objective: To examine the histological lesions associated with equine asthma during disease exacerbation and remission, and their relationship with lung function and BALF neutrophilia. Methods: Observational retrospective study. Methods: Pe...
Murcia RY, Vargas A, Lavoie JP.Glucocorticoids (GCs) are the most effective drugs for the treatment of human asthma. However, a subgroup of asthmatic patients with neutrophilic airway inflammation is insensitive to GCs. Interleukin-17 (IL-17), a cytokine upregulated in the airways of a subset of human asthmatic patients, contributes to the recruitment of neutrophils and induces a glucocorticoid resistance in human airway epithelial cells. We hypothesized that IL-17 similarly activates neutrophils and contributes to their persistence in the asthmatic airways in spite of glucocorticoid therapy. To determine whether IL-17 dire...
Rebordão MR, Amaral A, Lukasik K, Szóstek-Mioduchowska A, Pinto-Bravo P, Galvão A, Skarzynski DJ, Ferreira-Dias G.Prostaglandin E (PGE) has contradictory effects in many organs. It may have proinflammatory, anti-inflammatory, or anti-fibrotic roles, depending on the type of receptors to which it binds. By signaling through its receptors EP2 and EP4, PGE mediates anti-inflammatory and anti-fibrotic actions. In spite of chronic endometrial fibrosis (endometrosis) being a major cause of mare infertility, its pathogenesis is not fully understood. We have shown that contact of mare endometrium in vitro with neutrophil extracellular traps (NETs) proteases favors endometrial collagen type I production. Therefor...
Troedsson MH, Desvousges A, Alghamdi AS, Dahms B, Dow CA, Hayna J, Valesco R, Collahan PT, Macpherson ML, Pozor M, Buhi WC.Seminal plasma has been suggested to be involved in sperm transport, and as a modulator of sperm-induced inflammation, which is thought to be an important part of sperm elimination from the female reproductive tract. This article reports on recent experiments on the importance of seminal plasma components in sperm transport and elimination. In Experiment 1, hysteroscopic insemination in the presence (n = 3) or absence (n = 3) of 2 ng/mL PGE showed an increased portion of spermatozoa crossing the utero-tubal junction in the presence of PGE in two mares, while no difference was observed between ...
Derochette S, Franck T, Mouithys-Mickalad A, Ceusters J, Deby-Dupont G, Lejeune JP, Neven P, Serteyn D.In neutrophils (PMNs), superoxide anion (O2*-), the first reactive oxygen species (ROS) produced to kill pathogenic agents, is generated by NADPH oxidase, an enzymatic complex formed by the translocation of cytosolic subunits to the membrane flavocytochrome b558. In horses, excessive activation of PMNs is often associated with deadly pathologies and the modulation of their ROS production by acting on NADPH oxidase is a prime target to manage inflammation. We developed a cell-free assay to measure the activity of equine NADPH oxidase assembled in vitro, in order to test the effects of natural o...
Sheats MK, Pescosolido KC, Hefner EM, Sung EJ, Adler KB, Jones SL.Neutrophil infiltration is a prominent feature in a number of pathologic conditions affecting horses including recurrent airway obstruction, ischemia-reperfusion injury, and laminitis. Cell signaling components involved in neutrophil migration represent targets for novel anti-inflammatory therapies. In order to migrate into tissue, neutrophils must respond to chemoattractant signals in their external environment through activation of adhesion receptors (i.e. integrins) and reorganization of the actin cytoskeleton. Myristoylated Alanine-Rich C-Kinase Substrate (MARCKS), a highly conserved actin...
Massoco C, Palermo-Neto J.Benzodiazepines (BDZ) are among the most frequently used class of psychotropic drugs employed in veterinary medicine in Brazil and worldwide due to their anxiolytic, muscle relaxant and anticonvulsant effects [J. Clin. Pharmacol. 33 (1993) 124]. Peripheral benzodiazepine receptor (PBR) sites were described in peripheral organs, endocrine steroidogenic tissues and immune organs and cells. Midazolam is a mixed-type agonist of PBRs. The present study is focused on the effects of midazolam on equine peripheral blood neutrophils, peritoneal macrophages and cortisol levels in plasma. Adult horses we...
Perez B, Henriquez C, Sarmiento J, Morales N, Folch H, Galesio JS, Uberti B, Morán G.Neutrophilic asthma is an important disease subgroup, including patients with severe phenotypes and erratic responses to standard treatments. Tamoxifen (TX), a selective estrogen receptor modulator (SERM) used as treatment of human breast cancer, has been shown to induce early apoptosis of equine blood and bronchoalveolar lavage fluid (BALF) neutrophils in vitro. Equine recurrent airway obstruction (RAO) is a naturally occurring neutrophilic condition, closely related with human asthma. Our purpose was to investigate the therapeutic potential of tamoxifen in horses with neutrophilic lung infla...
Couto MA, Harwig SS, Cullor JS, Hughes JP, Lehrer RI.Endogenous, cysteine-rich antimicrobial peptides known as defensins are prominent components of human, rabbit, and rat neutrophils, yet little is known about their occurrence in other mammalian species. Although we did not detect mature (i.e., processed) defensins in equine neutrophil granules, we found that these granules contained small amounts of other cysteine-rich peptides with antimicrobial activity. One of these, eNAP-1, was purified by a combination of gel permeation and reversed-phase high-performance liquid chromatography from acid extracts prepared from the cytoplasmic granules of e...
Tremblay GM, Ferland C, Lapointe JM, Vrins A, Lavoie JP, Cormier Y.Bronchoalveolar lavages (BAL) were performed before and after 3 weeks of housing in 5 horses suffering from COPD and 5 normal horses. In the two groups, the total number of cells recovered remained unchanged after stabling. The most common cell populations in BAL fluid of control animals were alveolar macrophages (46.4%) and lymphocytes (44.9%). The percentage of neutrophils increased after stabling from 8.7% to 27.6%. In COPD horses, lymphocytes predominated (40.7%) in animals at pasture with neutrophils increasing from 29.4% to 71.6% after stabling. After fractionation by Percoll density gra...
Nolen-Walston RD, Harris M, Agnew ME, Martin BB, Reef VB, Boston RC, Davidson EJ.To determine whether there are important differences relating to seasonality of signs or clinical features between subtypes of inflammatory airway disease (IAD) in horses caused by neutrophilic and eosinophilic-mastocytic inflammation having dissimilar etiopathologic pathways. Methods: Retrospective case series. Methods: 98 horses. Methods: Data were compiled from medical records of horses examined because of poor performance from 2004 through 2010. Horses underwent a standardized high-speed treadmill test, lameness evaluation, cardiac evaluation, and postexercise bronchoalveolar lavage (BAL)....
Uberti B, Morán G.Neutrophilic bronchiolitis is the primary lesion in asthma-affected horses. Neutrophils are key actors in host defense, migrating toward sites of inflammation and infection, where they act as early responder cells toward external insults. However, neutrophils can also mediate tissue damage in various non-infectious inflammatory processes. Within the airways, these cells likely contribute to bronchoconstriction, mucus hypersecretion, and pulmonary remodeling by releasing pro-inflammatory mediators, including the cytokines interleukin (IL)-8 and IL-17, neutrophil elastase, reactive oxygen specie...
Doty A, Buhi WC, Benson S, Scoggin KE, Pozor M, Macpherson M, Mutz M, Troedsson MH.Equine spermatozoa induce a uterine inflammatory response characterized by a rapid, transient influx of polymorphonuclear neutrophils (PMNs). Seminal plasma proteins have been shown to modulate the interaction between spermatozoa and PMNs, but a specific protein responsible for this function has not been identified. The objective of this study was to isolate and identify a protein in equine seminal plasma that suppresses binding between spermatozoa and PMNs. Seminal plasma was pooled from five stallions, and proteins were precipitated in 60% (w/v) ammonium sulfate and dialyzed (3500 MW cutoff)...
Gy C, Leclere M, Vargas A, Grimes C, Lavoie JP.Asthma in horses is associated with nonspecific respiratory clinical signs and may be manifested only as exercise intolerance. Its diagnosis relies on bronchoalveolar lavage fluid (BALF) cytology in the presence of compatible clinical signs. The identification of blood biomarkers for this condition would facilitate diagnosis in the field, because there are regional areas where BAL is not routinely performed in clinical practice. Objective: Identification of blood biomarkers for the diagnosis of asthma in horses. Methods: Fourteen horses with asthma with increased neutrophil numbers in BALF (ne...
Aragona F, Tabbì M, Gugliandolo E, Giannetto C, D'Angelo F, Fazio F, Interlandi C.Cannabidiol (CBD) is a non-psychotropic cannabinoid obtained from hemp (Cannabis sativa L.) used for pain management in companion animals including horses. The present study aimed to evaluate the efficacy of cannabidiolic acid (CBDA) and cannabigerol/cannabidiol oil (CBG/CBD) oral administration in alleviating pain in adult horses affected by chronic osteoarthritis (OA). Twenty-four horses (10 geldings and 14 mares), aged between 11 and 18 years old, were equally divided into two groups. One group received CBDA 15% oil and the other group received CBG/CBD oil (CBG20%-CBD10%) for 14 consecuti...
Fogarty U, Buckley T.Significant differences were detected by bronchoalveolar lavage (BAL) between horses racing successfully and those showing exercise intolerance. Neutrophil percentage, haemosiderophage percentage and total bacterial numbers were significantly elevated in horses with exercise intolerance. BAL provided a more accurate indication of the incidence and extent of exercise induced pulmonary haemorrhage (EIPH) than visual inspection.
Troedsson MH, Woodward EM.Equine endometritis is characterized by the expression of pro- and anti- inflammatory cytokines and an influx of polymorphonuclear neutrophils (PMNs) into the uterus. Mares resistant to persistent endometritis clear the inflammation within 24-36 h after exposure to microorganisms or semen. These mares have a rapid increase of pro-inflammatory cytokines and an upregulation of inflammatory modulating cytokines within 6h after exposure to inflammatory challenge. In conjunction with effective uterine contractions, these events are believed to be responsible for the transient nature of the inflamma...
Franck T, Grulke S, Deby-Dupont G, Deby C, Duvivier H, Peters F, Serteyn D.Equine inflammatory disease is accompanied by a neutrophil activation resulting in the release of granulocytic enzyme myeloperoxidase (MPO). To measure MPO in horse plasma as marker of neutrophil activation, the authors purified equine neutrophil MPO and developed a specific enzyme immunoassay using 2 specific polyclonal antibodies obtained from rabbit (primary antibody) and guinea pig (secondary antibody). The sandwich complex "primary antibody-MPO-secondary antibody" was detected using a goat anti-guinea pig immunoglobulin antibody conjugated to alkaline phosphatase. The enzyme-linked immuno...
Janssen P, Tosi I, Hego A, Maréchal P, Marichal T, Radermecker C.Asthma encompasses a spectrum of heterogenous immune-mediated respiratory disorders sharing a similar clinical pattern characterized by cough, wheeze and exercise intolerance. In horses, equine asthma can be subdivided into severe or moderate asthma according to clinical symptoms and the extent of airway neutrophilic inflammation. While severe asthmatic horses are characterized by an elevated neutrophilic inflammation of the lower airways, cough, dyspnea at rest and high mucus secretion, horses with moderate asthma show a milder neutrophilic inflammation, exhibit intolerance to exercise but no...
Pirie RS, Collie DD, Dixon PM, McGorum BC.To evaluate inhaled hay dust suspensions (HDS) as a tool for the diagnosis and investigation of heaves, the pulmonary inflammatory and functional consequences of inhalation challenge with 3 different HDS were determined in 6 control and 7 asymptomatic heaves horses. Heaves horses given HDS challenge developed the characteristic features of heaves, including airway neutrophilia, obstructive airway dysfunction and mucus hypersecretion. While HDS challenge induced a mild airway neutrophilia in controls, the no-response threshold for controls was greater than that of heaves horses, and there was n...
Moore BR, Krakowka S, Robertson JT, Cummins JM.Cytologic examination of bronchoalveolar lavage fluid (BALF), including phenotypic analysis of lymphocytes, was performed on 32 Standardbreds with poor race performance and endoscopic examination findings characteristic of inflammatory airway disease (IAD). Nucleated cell counts in BALF from IAD-affected horses were higher than those in control horses; the cytologic profile of BALF in affected horses included mixed inflammation, characterized by mild neutrophilia, lymphocytosis, and monocytosis. Eosinophil and mast cell counts were not higher in the IAD-affected group, compared with those in t...
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...
Leise B.Equine laminitis is a devastating disease in which failure of the adhesion between the digital dermal and epidermal laminae at the basement membrane results in crippling lameness and structural damage to the foot of the horse. Laminitis occurring secondary to sepsis is known to result from a significant inflammatory response that includes leukocyte emigration into the lamellar tissue. These leukocytes, in particular the neutrophil, have been extensively evaluated in experimental models of sepsis-related laminitis in the horse. This review will discuss the relevant findings elucidated from thes...
Sweeney CR, Rossier Y, Ziemer EL, Lindborg S.Bronchoalveolar lavage (BAL) fluid was analyzed in healthy horses, using different lavage fluid volumes and lung sites. The only significant difference in the cellular composition of BAL fluid between the right and left lungs was the mast cell numbers, which were significantly higher in the left lung. Total cell count ranged from 34 to 330 cells/microliter for the right lung and 43 to 330 cells/microliter for the left lung. Percentage of neutrophils ranged from 1 to 7% in the right lung and 1 to 5% in the left lung. The small-volume (50 ml) lavage had a greater percentage of neutrophils and a ...
Derksen FJ, Scott JS, Miller DC, Slocombe RF, Robinson NE.We performed bronchoalveolar lavage (BAL) in 2 groups of ponies. Principal ponies had a history of heaves, a disease characterized by recurrent airway obstruction and airway hyperreactivity when ponies are housed in a barn and fed hay; control ponies had no history of airway obstruction. Ponies were paired (principal and control), and BAL was performed after 2 months of being pastured when principal ponies were in clinical remission (Period A), after barn housing when principal ponies had acute airway obstruction (Period B), and after a 1- and a 2-wk recovery phase of pasture grazing (Periods ...
Sanchez-Teran AF, Bracamonte JL, Hendrick S, Riddell L, Musil K, Hoff B, Rubio-Martínez LM.The objective of this study was to evaluate the effect of through-and-through joint lavage on systemic and synovial serum amyloid A (SAA), total protein, nucleated cell count and percentage of neutrophils in the synovial fluid of six healthy horses. A prospective experimental study was performed where one healthy tarsocrural joint of each horse was randomly assigned to receive repeated through-and-through joint lavage at 0, 48 and 96 h. Synovial fluid and blood samples were collected at 0 (baseline), 24, 48, 72, 96 and 120 h. Systemic and synovial SAA, total protein, nucleated cell count a...
Mateo-Otero Y, Zambrano F, Catalán J, Sánchez R, Yeste M, Miro J, Fernandez-Fuertes B.In several mammalian species, acute endometritis driven by the recruitment of polymorphonuclear cells (PMN) occurs in response to semen. These PMNs release DNA to form neutrophil extracellular traps (NETs) in cattle, horse and human, leading to sperm entrapment. While there is no evidence of this phenomenon occurring in donkeys, artificial insemination (AI) with frozen-thawed semen, which results in very poor pregnancy rates, leads to a large PMN recruitment to the uterus. Objective: To investigate whether donkey semen can trigger NET release (NETosis) and if excessive NETosis occurs in respon...
Niedzwiedz A, Jaworski Z, Tykalowski B, Smialek M.Dysregulation of apoptosis has been implicated in a range of diseases including tumors, neurodegenerative and autoimmune diseases, as well as allergic asthma and chronic obstructive pulmonary disease (COPD) in humans. Although it has a different pathophysiology, delayed apoptosis of various inflammatory cells may play a pivotal role in the development of recurrent airway obstruction (RAO) in horses. Reduction of inflammatory cell apoptosis or a dysregulation of this process could lead to chronic inflammation and tissue injury. Therefore, the aim of this study was to investigate the rate of apo...
