Phagocytosis in horses refers to the process by which certain cells, known as phagocytes, engulf and digest foreign particles, bacteria, and cellular debris. This cellular mechanism is a part of the innate immune response, helping to protect horses from infections and maintain tissue homeostasis. Key phagocytic cells in horses include neutrophils and macrophages, which are responsible for identifying and eliminating pathogens. Research on phagocytosis in equines explores the efficiency, regulation, and impact of this process on overall health and disease resistance. This page includes peer-reviewed research studies and scholarly articles that investigate the mechanisms, cellular interactions, and implications of phagocytosis in equine immunology.
Theoret CL, Barber SM, Moyana T, Townsend HG, Archer JF.The reparative ability of equine synovium was determined by gross, histological, and ultrastructural examination. The functional potential of the synovium was estimated by examination of synovial cell organelles with transmission electron microscopy. Results from rested and exercised horses were compared to determine the effect of exercise on synovial healing. The response of synovectomized joint to exercise was evaluated with a standardized lameness examination and by gross, histological, and histochemical observations of the articular cartilage. A 7-mm diameter motorized synovial resector wa...
Madigan JE, Valberg SJ, Ragle C, Moody JL.Severe muscle cramping not associated with exercise was observed in 5 horses. Focal muscle groups in various regions underwent intermittent visible contraction. Intermittent prolapse of the third eyelid, sweating, pawing, muscle tremors, and muscle fasciculations also were observed. Clinical signs often were misconstrued as signs of colic. Percussion of muscle induced contraction of muscle groups. Concentrations of serum electrolytes and the acid-base balance were within reference limits, but activities of creatine kinase and aspartate transaminase were moderately high. Muscle biopsy revealed ...
Causey RC, Paccamonti DL, Todd WJ.A total of 22 clinical streptococcal isolates, predominantly Streptococcus zooepidemicus, associated with endometritis in horses were tested for their ability to withstand the natural bactericidal properties of freshly obtained blood. During a 3-hour incubation in blood from a single horse, 8 of these isolates survived and grew, the remainder were killed. To determine whether this ability to grow extended to blood of other horses, 5 of these growing isolates were tested for their ability to grow in the blood of 5 additional horses. The same 5 horses were used for each isolate. The isolates gre...
López Alvarez ML, Bustos Obregón E.The occurrence of phagocytosis of spermatozoa in the epididymis is a controversial issue in male reproductive biology. In this paper we documented, by scanning and electron microscopy, the presence of spermatophages in the lumen of the proximal region of stallion epididymis. Sperm phagocytosis involving the principal cells that line the cauda epididymidis region was also observed. The possible meaning of this process in the removal of damaged, aged or surplus spermatozoa is discussed. These observations were made in samples obtained from random cases, during the course of research work carried...
Johannisson A, Gröndahl G, Demmers S, Jensen-Waern M.Methodological aspects of flow-cytometric evaluation of the phagocytic properties of equine neutrophils were elucidated. The kinetics of attachment and ingestion were studied, and the phagocytic process was more rapidly completed when serum-opsonized yeast cells were used than with use of IgG-opsonized yeast cells. Trypan blue was successfully used to quench fluorescence of non-ingested yeast cells. There were only minor differences in the kinetics of phagocytosis between quenched and unquenched samples, indicating that attachment is rapidly followed by ingestion. Trypan blue quenching caused ...
Mazet I, Pendland J, Boucias D.In this study, the phagocytic ability of Spodoptera exigua hemocytes was compared to horse neutrophils. In vitro assays showed that the insect granulocytes and horse neutrophils actively phagocytose FITC-labeled Paecilomyces farinosus blastospores opsonized with S. exigua hemolymph lectin or horse serum, respectively. Killing of fungal cells by the neutrophils and hemocytes was analyzed under in vitro conditions. Neutrophils reduced the growth of P. farinosus up to 65% whereas no fungicidal activity was observed with hemocyte monolayers. The production of oxygen metabolites by both phagocytic ...
Hondalus MK, Mosser DM.Rhodococcus equi is a facultative intracellular bacterium of macrophages that can cause serious pneumonia in both young horses and immunocompromised people. Essential to understanding rhodococcus pathogenesis is a quantitative documentation of the intracellular events that follow macrophage phagocytosis of the organism. By using a bacterial immunofluorescence staining assay, we verified the intracellular survival and replicative potential of R. equi in both murine peritoneal macrophages and equine alveolar macrophages in vitro. Following an initial lag period of 6 to 12 h, the intracellular nu...
Kluciński W, Winnicka A, Olszewski M, Sikora J, Sitarska E, Niemiałtowski M, Muzylak M, Bylinka G, Wyszyński M.The aim of this study was to compare phagocytic activity of polymorphonuclear cells (PMNs) from the bronchoalveolar lavage of clinically healthy horses and those with severe chronic bronchiolitis. Research was carried out on 28 horses. Chronic inflammation of the lower airways was diagnosed in nine horses. Cells from the respiratory tract were lavaged according to accepted methods. For comparison, PMNs were isolated from peripheral blood of all investigated horses. The phagocytic activity of PMNs was determined in relation to two standard strains of Staphylococcus aureus, Staph, aureus Smith w...
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...
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, ...
Messick JB, Rikihisa Y.The binding, internalization, and proliferation of Ehrlichia risticii in P388D1 cells and equine polymorphonuclear (PMN) leukocytes were studied by immunofluorescent staining and flow cytometric analysis. The binding of ehrlichiae to P388D1 cells at 4 degrees C was dose dependent, and the antigens of bound organisms were susceptible to pronase treatment. Additionally, the binding of ehrlichiae to P388D1 cells was diminished when either P388D1 cells or ehrlichiae were treated with 1% paraformaldehyde for 30 min or 0.25% trypsin for 15 min. These results indicate that the ehrlichial ligand and h...
Atwal OS, Singh B, Staempfli H, Minhas K.The pulmonary intravascular macrophages (PIMs) have been described in several species of animals. This study demonstrates for the first time that the equine lung has PIMs as resident phagocytes in its microvasculature. Their salient features such as globular surface coat, structures of the endocytic pathway, and related cell organelles closely resemble those of the calf, goat, and sheep. The exquisite organization of the coat globules in the form of a linear chain was structurally similar to the lipolytic lipase and the heparin-sensitive globular coat from PIMs of calf, goat, and sheep. Monast...
Crisman MV, Hodgson DR, Bayly WM, Liggitt HD.To determine whether road transport affected pulmonary phagocyte activity, 7 healthy Thoroughbred horses were shipped 1,160 kilometers over 36 hours. Fluid collected by bronchoalveolar lavage (BAL) 12 hours, and 7 and 14 days after transport was analyzed. Results were compared to those from the same horses pre-transport, and 7 non-transported control horses that had BAL performed at the same times as the transported horses. Of cells recovered with BAL the percentage of viable pulmonary alveolar macrophages (PAMs) declined from 90.0 +/- 0.9% pre-transport to 80.0 +/- 3.7% by 2 weeks post transp...
Ishino S, Kumagai K, Kuniyoshi S, Nakazawa M, Matsuda I, Oka M.An immunohistochemical analysis of Rhodococcus equi-induced pneumonia in 10 foals was performed by biotin-streptavidin system. The detection of R. equi was more sensitive in immuno-stain using anti-R. equi serum than in Gram's stain. This bacteria also reacted to anti-BCG serum. Lysozyme and alpha 1-antitrypsin were detectable in macrophages. A particularly intense staining was observed in association with intracellular bacteria. Though a degree of reaction for alpha 1-antichymotrypsin was very low in comparison with lysozyme and alpha 1-antitrypsin, it was also demonstrated in macrophages ing...
Daniel GB, Tucker RL, Buckman T, Daniel SL.Isolated equine granulocytes (WBC), radiolabeled with 99mTc-hexamethylpropyleneamine oxime (99mTc-HMPAO) or 111In-oxine, were evaluated in vitro for their labeling characteristics, viability, and phagocytic function over a 6-hour postlabeling period. Mean +/- SD labeling efficiency for 111In-oxine-WBC was 62.2 +/- 15.3%, which was significantly (P less than 0.001) higher than that for 99mTc-HMPAO-WBC (32.0 +/- 17.0%). In vitro elution of radiolabel from cells was significantly (P less than 0.02) greater for 99mTc-HMPAO-WBC at 0.5, 2, and 4 hours, but was not significantly different from elutio...
Hago BE, Vaughan LC, Plummer JM.A transmission electron microscope study was undertaken to investigate the details of the synovial tendon sheath and bursal lining in horses. The lining cells appeared to be fibroblasts and were buried in a finely granular ground substance. Generally these cells had poor cytoplasmic organelles, sparse short profiles of rough endoplasmic reticulum (RER), few scattered ribosomes and, occasionally, a poorly developed Golgi complex. However, a few lining cells appeared more active, having pronounced RER with dilated cisternane. The surface of the lining cells, particularly those with dilated RER, ...
Wichtel MG, Anderson KL, Johnson TV, Nathan U, Smith L.Functional activities (phagocytosis and killing) of neutrophil leucocytes (NL) and immunoglobulin G concentrations were evaluated in six healthy foals from birth to 6 months of age. Peripheral blood NL were reacted with Streptococcus equisimilis in 20 per cent pooled equine serum for 30, 60 and 90 mins and functional activities of NL were determined using a fluorochrome microassay. Values for foal NL function were compared with those of healthy adult horses (n = 28). Foal neutrophil function was influenced by age. Killing capacity of NL decreased, whereas phagocytic capacity increased, until 1...
Buschmann H, Baumann M.During strenuous exercise of horses that are prepared for international Three-Day-Events a significant decrease in the in vitro killing rate of phagocytosed yeast cells by the blood granulocytes has been observed. Other immunological parameters, such as the phorbolmyristate dependent chemiluminescence in granulocytes and the mitogenic stimulation of blood lymphocytes, remained unchanged.
Bogdan I, Nechifor A, Băşea I, Hruban E.In the Rumanian traditional medicine a transcutaneous implantation of the root of Helleborus purpurascens is used to provoke leucocytosis and neutrophils with the aim to activate chronic diseases for better healing. The skin is perforated with a thick needle (in cattle and horses in the area of the thorax, in sheep and pigs in the ear flap), the transplant is introduced and after 24 hours removed. Though the method is more than 100 years old, it is not reported in the literature. The way of action is unknown. The investigation are performed to show the efficacy of this method of Helleborus-imp...
Foerster RJ, Wolf G.Equine blood polymorphonuclear leukocytes (PMN) were isolated by buffy coat and hypotonic lysis of residual erythrocytes. A highly reproducible method is described for measuring the uptake of opsonized latex microspheres by equine PMN using flowcytometry. The use of cytochalasin D allowed for differentiation of ingested from attached particles. The kinetics of phagocytosis in vitro is shown for different experimental conditions. We developed an assay for evaluation of phagocytic capacity of PMN which allows the assessment of drugs for their influence on phagocytosis in vivo as well as in vitro...
Jain NC, Vegad JL, Kono CS.Equine neutrophil antibody was raised in rabbits inoculated with equine neutrophils isolated to purity greater than 99.0%, using Percoll density-gradient sedimentation. Neutrophil antibody was detected by use of agar gel diffusion, leukoagglutination, indirect immunofluorescence, staphylococcal protein A and streptococcal protein G binding, and phagocytic inhibition techniques. Precipitin lines and leukoagglutination were seen in antiserum dilutions of 1:4 and 1:64, respectively. The specific nature of leukoagglutination was characterized by the formation of rosette-like clumps of neutrophils....
Troedsson M, Concha C, Einarsson S, Holmberg O.From 6 mares with chronic uterine infection, polymorphonuclear neutrophils (PMNs) were obtained from the uterus. In order to recover an adequate number of viable PMNs, 0.1% oyster glycogen was infused into the uterus as a mild irritant 12 h prior to the uterine flushing. Phagocytosis and chemotaxis of the uterine derived PMNs were determined. The supernatant from the uterine flushing was compared with autologous serum for its capacity as an opsonin and chemoattractant. There was a significant increase of both phagocytosis and chemotaxis when autologous serum was used compared with the supernat...
Wong CW, Thompson HL, Thong YH, Thornton JR.Bronchoalveolar lavage samples were collected using a fibreoptic endoscope from horses at specified times before and after single bouts of exercise. Lucigenin-dependent phagocytic chemiluminescence was used to assess the effect of exercise on the alveolar macrophage metabolic activity in response to stimulation by opsonised zymosan. A profound suppressive effect on the chemiluminescence production was present throughout the first three days after exercise. However, the cellular composition of lavage fluids was not altered by the exercise. It is suggested that strenuous exercise may jeopardize ...
Blancquaert AB, Colgan SP, Bruyninckx WJ.To identify the influence of technical factors on the in vitro motility of equine neutrophils towards streptococcus culture supernatant in an under-agarose assay, we studied the changes in eight cell migration parameters. The distances the phagocytes travelled by directed, random and spontaneous migration increased with incubation time, cell concentration and the gelatin and serum contents of the migration plates. The contribution of chemotaxis to the phagocyte migrations, however, decreased simultaneously. The directed and random, though not the spontaneous, migrations of the phagocytes incre...
Morris DD, Harmon BG, Moore JN.A study was performed to determine whether equine antiserum to core lipopolysaccharide (LPS) would enhance phagocytosis of smooth gram-negative (GN) organisms by equine macrophages. Five healthy adult horses (group A) were immunized with a bacterin prepared from the J-5 mutant of Escherichia coli 0111:B4 and Salmonella minnesota R595 to produce antibodies to core LPS. Five horses (group B) served as nonimmunized controls and were given physiologic saline solution instead of the rough mutant bacterin. Serum antibody titers to core LPS and to smooth E coli 0111:B4 were determined by indirect ELI...
Dennis VA, Klei TR, Chapman MR, Jeffers GW.Eosinophils and neutrophils from ponies with Strongylus vulgaris-induced eosinophilia (eosinophilic ponies; activated eosinophils and neutrophils) were assayed in vitro for chemotactic and chemokinetic responses to zymosan-activated serum (ZAS) using the filter system in Boyden chambers, for Fc and complement (C) receptors using the EA and EAC-rosette assays, respectively, and for phagocytic and bactericidal activities using opsonized Escherichia coli and the acridine orange method. The responses of activated eosinophils and neutrophils in the above assays were compared with those of eosinophi...
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.
Blancquaert AM, Colgan SP, Bruyninckx WJ.Although equine neutrophils did not respond towards formylated methionyl peptides, Streptococcus faecalis culture supernatant caused an in vitro stimulation of equine neutrophil motility when measured by an under-agarose assay. The migration of neutrophils towards the culture supernatant increased sigmoidally with the logarithmic concentration of the culture supernatant in the chemoattractant wells. The streptococcal culture supernatant was chemokinetic because it stimulated the random motility of the phagocytes. Because granulocytes migrated further towards the supernatant than could be expla...
Traub-Dargatz JL, McKinnon AO, Bruyninckx WJ, Thrall MA, Jones RL, Blancquaert AM.Four bronchoalveolar lavages were performed sequentially on 9 control and 8 transport-stressed female horses. Alterations in results of fluid cytologic analyses, microbial content, and phagocyte function of recovered pulmonary macrophages in all horses were determined. Seemingly, absolute and relative increase in the number of inflammatory cells detected in the second bronchoalveolar lavage fluid of control horses was the result of irritation of the first lavage. This increased response was not observed in transport-stressed horses until 5 days after transport (third lavage; 10 days after init...
Morris DD, Strzemienski PJ, Gaulin G, Spencer P.Neutrophil function was evaluated in six clinically normal adult horses, immediately before and 3-6 hours after they were given one dose of hydrocortisone sodium succinate (1 mg/kg body weight). Random migration, stimulated migration to zymosan-activated serum, bacterial phagocytosis and bactericidal capacity of neutrophils were determined in vitro. The mean indices of stimulated migration (net migration and migration ratio) were significantly greater after CS administration (net migration = 62 +/- 23 micron; migration ratio = 11.5 +/- 6.7) than before CS administration (net migration = 44 +/-...
Ganjam VK, McLeod C, Klesius PH, Washburn SM, Kwapien R, Brown B, Fazeli MH.The reaction between ovarian hormones and experimental uterine infection (Streptococcus zooepidemicus) was investigated in 3 groups, each containing 6 ovariectomized mares. Group 1 served as controls ('anoestrus'), Group 2 mares were injected with oestrogen ('oestrus') and Group 3 with progesterone ('dioestrus') over a period of 5 weeks. All mares received an intrauterine inoculation of the bacteria 1 week after the start of hormonal treatment, and the results of the challenge were examined by endometrial biopsy and swabs once weekly. At the end of Week 1 no bacteria were recovered from the ma...
Watson ED, Stokes CR, Bourne FJ.Both random and directional migration of blood neutrophils from 9 mares susceptible to persistent endometritis were significantly less (p less than 0.05) than neutrophils from 8 resistant mares. Serum from susceptible mares had significantly more (p less than 0.01) chemotactic activity than serum from resistant mares. Although phagocytosis of yeast blastospores by blood neutrophils from 4 resistant and 3 susceptible mares was similar, uterine neutrophils from susceptible mares were significantly worse (p less than 0.01) at phagocytosis than uterine neutrophils from resistant mares. Uterine was...
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 ...
Raidal SL, Bailey GD, Love DN.Flow cytometry was used to assess the phagocytosis of fluorescent-labelled bacteria by equine peripheral blood neutrophils and pulmonary alveolar macrophages. Cell populations were prepared from venous blood following ammonium chloride lysis and from washed bronchoalveolar lavage derived samples. Discrete clusters of cells, corresponding to different leucocyte groups, were readily identified on the basis of differing light scattering properties and could thus be discriminated, negating the need for prior cell separation. Cells able to associate with fluorescent-labelled bacteria (by attachment...
Traub-Dargatz JL, McKinnon AO, Bruyninckx WJ, Thrall MA, Jones RL, Blancquaert AM.Four bronchoalveolar lavages were performed sequentially on 9 control and 8 transport-stressed female horses. Alterations in results of fluid cytologic analyses, microbial content, and phagocyte function of recovered pulmonary macrophages in all horses were determined. Seemingly, absolute and relative increase in the number of inflammatory cells detected in the second bronchoalveolar lavage fluid of control horses was the result of irritation of the first lavage. This increased response was not observed in transport-stressed horses until 5 days after transport (third lavage; 10 days after init...
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.
Caston SS, Cooper EE, Chandramani-Shivalingappa P, Sponseller BA, Hostetter JM, Sun Y.We investigated CD47 expression in cryopreserved sections of equine cutaneous masses and normal skin. CD47 is a cell surface protein expressed on many cell types and overexpressed in some tumors. Interaction of CD47 and signal regulatory protein-alpha (SIRPα) inhibits phagocytosis by macrophages. Formalin-fixed tissues from horses prospectively enrolled in the study were used to establish a histologic diagnosis. Immunohistochemical assays were performed on cryopreserved tissues using anti-CD47 antibodies or IgG control antibodies. CD47 was not expressed on equine normal skin but positivity to...
López Alvarez ML, Bustos Obregón E.The occurrence of phagocytosis of spermatozoa in the epididymis is a controversial issue in male reproductive biology. In this paper we documented, by scanning and electron microscopy, the presence of spermatophages in the lumen of the proximal region of stallion epididymis. Sperm phagocytosis involving the principal cells that line the cauda epididymidis region was also observed. The possible meaning of this process in the removal of damaged, aged or surplus spermatozoa is discussed. These observations were made in samples obtained from random cases, during the course of research work carried...
Wadsworth AB, Sickles GM.IT HAS BEEN SHOWN THAT THE PNEUMOCOCCUS MULTIPLYING IN THE TISSUES OF THE IMMUNIZED ANIMAL (HORSE) BECOMES ATTENUATED: loses, in varying degrees, its virulence, capacity of capsule formation, susceptibility to phagocytosis, and type specificity. The antigenic activity as an immunizing agent and the production of "soluble specific substance" are also altered. In some instances, the typical pneumococcus characteristics may be quickly restored by one or two passages through a susceptible animal (mouse). In others, virulence is not recovered and the organism remains atypical. Whether these changes...
Buschmann H, Baumann M.During strenuous exercise of horses that are prepared for international Three-Day-Events a significant decrease in the in vitro killing rate of phagocytosed yeast cells by the blood granulocytes has been observed. Other immunological parameters, such as the phorbolmyristate dependent chemiluminescence in granulocytes and the mitogenic stimulation of blood lymphocytes, remained unchanged.
Alghamdi AS, Fedorka CE, Scoggin KE, Esteller-Vico A, Beatty K, Davolli G, Ball BA, Troedsson MHT.Sperm-neutrophil binding is an important facet of breeding and significantly impacts fertility. While a specific seminal plasma protein has been found to reduce this binding and improve fertility (CRISP-3), additional molecule(s) appear to promote binding between defective sperm and neutrophils. Recent work has suggested one of these proteins is lactoferrin (LF), an 80 kDa iron-binding protein found throughout the body, but the purity of the protein was not confirmed. It is unknown if LF binds to sperm selectively based on viability, and if receptors for LF are located on equine sperm. To eval...
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-...
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...
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...
Jain NC, Vegad JL, Kono CS.Equine neutrophil antibody was raised in rabbits inoculated with equine neutrophils isolated to purity greater than 99.0%, using Percoll density-gradient sedimentation. Neutrophil antibody was detected by use of agar gel diffusion, leukoagglutination, indirect immunofluorescence, staphylococcal protein A and streptococcal protein G binding, and phagocytic inhibition techniques. Precipitin lines and leukoagglutination were seen in antiserum dilutions of 1:4 and 1:64, respectively. The specific nature of leukoagglutination was characterized by the formation of rosette-like clumps of neutrophils....
Imai S, Ozeki K, Fujita J.The surface structure of the ciliary zone in 13 species of ciliates found in the large intestine of the horse was observed by scanning electron microscopy. In Holophryoides ovalis many fine depressions considered to be a result of phagocytosis or pinocytosis in the naked cytostome were noticed. In Blepharocorys spp. a distinct section was present between the portion with cilia and that without cilia. It was not present, however, in some species of the family Buetschliidae, such as Bundleia postciliata and Didesmis spp. The species of Entodiniomorphida had a lip around the ciliary zone with cil...
Vanderstock JM, Lecours MP, Lavoie-Lamoureux A, Gottschalk M, Segura M, Lavoie JP, Jean D.OBJECTIVE To evaluate in vitro phagocytosis and bactericidal activity of circulating blood neutrophils in horses with severe equine asthma and control horses and to determine whether circulating blood neutrophils in horses with severe equine asthma have an increase in expression of the proinflammatory cytokine tumor necrosis factor (TNF)-α and the chemokine interleukin (IL)-8 and a decrease in expression of the anti-inflammatory cytokine IL-10 in response to bacteria. ANIMALS 6 horses with severe equine asthma and 6 control horses. PROCEDURES Circulating blood neutrophils were isolated from h...
Kluciński W, Winnicka A, Olszewski M, Sikora J, Sitarska E, Niemiałtowski M, Muzylak M, Bylinka G, Wyszyński M.The aim of this study was to compare phagocytic activity of polymorphonuclear cells (PMNs) from the bronchoalveolar lavage of clinically healthy horses and those with severe chronic bronchiolitis. Research was carried out on 28 horses. Chronic inflammation of the lower airways was diagnosed in nine horses. Cells from the respiratory tract were lavaged according to accepted methods. For comparison, PMNs were isolated from peripheral blood of all investigated horses. The phagocytic activity of PMNs was determined in relation to two standard strains of Staphylococcus aureus, Staph, aureus Smith w...
Ishino S, Kumagai K, Kuniyoshi S, Nakazawa M, Matsuda I, Oka M.An immunohistochemical analysis of Rhodococcus equi-induced pneumonia in 10 foals was performed by biotin-streptavidin system. The detection of R. equi was more sensitive in immuno-stain using anti-R. equi serum than in Gram's stain. This bacteria also reacted to anti-BCG serum. Lysozyme and alpha 1-antitrypsin were detectable in macrophages. A particularly intense staining was observed in association with intracellular bacteria. Though a degree of reaction for alpha 1-antichymotrypsin was very low in comparison with lysozyme and alpha 1-antitrypsin, it was also demonstrated in macrophages ing...
Bertram TA, Coignoul FL, Jensen AE.Equine neutrophils were combined with contagious equine metritis organism (CEMO) or Escherichia coli in vitro in the presence of seminal plasma, uterine flushings, or Hank's balanced salt solution (HBSS). Phagocytosis and intracellular killing were estimated by bacterial culture and light and electron microscopy. With lysed neutrophils, the numbers of colony-forming units of CEMO and E coli increased in seminal plasma and uterine flushings. Numbers of CEMO decreased in HBSS. The numbers of CEMO increased more in the presence of seminal plasma than the other media. When neutrophils were in the ...
Dennis VA, Klei TR, Chapman MR, Jeffers GW.Eosinophils and neutrophils from ponies with Strongylus vulgaris-induced eosinophilia (eosinophilic ponies; activated eosinophils and neutrophils) were assayed in vitro for chemotactic and chemokinetic responses to zymosan-activated serum (ZAS) using the filter system in Boyden chambers, for Fc and complement (C) receptors using the EA and EAC-rosette assays, respectively, and for phagocytic and bactericidal activities using opsonized Escherichia coli and the acridine orange method. The responses of activated eosinophils and neutrophils in the above assays were compared with those of eosinophi...
Morris DD, Harmon BG, Moore JN.A study was performed to determine whether equine antiserum to core lipopolysaccharide (LPS) would enhance phagocytosis of smooth gram-negative (GN) organisms by equine macrophages. Five healthy adult horses (group A) were immunized with a bacterin prepared from the J-5 mutant of Escherichia coli 0111:B4 and Salmonella minnesota R595 to produce antibodies to core LPS. Five horses (group B) served as nonimmunized controls and were given physiologic saline solution instead of the rough mutant bacterin. Serum antibody titers to core LPS and to smooth E coli 0111:B4 were determined by indirect ELI...
Blue MG, Brady AA, Davidson JN, Kenney RM.Forty uterine fluid samples were obtained during oestrus and dioestrus of successive cycles from 4 mares classified as resistant to bacterial infection of the uterus. The flushings were assayed for peroxidase activity and ability to influence phagocytosis by polymorphonuclear neutrophils. Uterine fluid volumes and protein concentrations and serum oestrogen values were similar during oestrus and dioestrus. Serum progesterone concentrations were depressed during oestrus. Equine neutrophils adherent to glass slides were exposed to a genital strain of Streptococcus zooepidemicus suspended in salin...
Salinas C, Espinosa G, Morales N, Henríquez C, Morán G, Gajardo G, Burgos RA, Uberti B.Obesity and insulin dysregulation (ID) are increasingly prevalent conditions in equid populations worldwide. Immune impairment is well described in humans with metabolic dysfunction and is reported but still incompletely understood in horses. This study evaluated the effect of acute induced transient hyperglycemia on apoptosis, phagocytosis and oxidative burst activity of peripheral blood polymorphonuclear cells (PMN) of lean and obese adult horses with or without insulin dysregulation. Seventeen adult horses were allocated into three groups based on their body condition score (BCS) and metabo...
Sternberg S, Johannisson A, Magnusson U, Jensen-Waern M.After exposure of equine granulocytes from both foals and adult horses to culture supernatants from clinical isolates of Actinobacillus equuli, phagocytic capacity and respiratory burst was examined by flow-cytometry and a chemiluminescence assay, respectively. One haemolytic isolate of an equine Actinobacillus was also included in the study. An average decrease of 22% in total number of granulocytes, in the flow cytometric assay (P < 0.01), and an average decrease of 26% in light emission, in the chemiluminescence assay (P < 0.001), was seen after exposure to bacterial culture supernata...
Allegro AR, Barhoumi R, Bordin AI, Bray JM, Cohen ND.Rhodococcus equi is a soil saprophytic bacterium and intracellular pathogen that causes pneumonia in foals. Strains of R. equi that are virulent in foals contain a plasmid that encodes a virulence-associated protein A (VapA) necessary for replication in macrophages. Because other intracellular pathogens survive and replicate inside amoebae, we postulated that the VapA-bearing plasmid (pVAPA) confers a survival advantage for R. equi against environmental predators like amoebae. To test this hypothesis, we compared phagocytosis by and survival in Acanthamoeba castellanii of isogenic strains of p...
Grondin TM, DeWitt SF, Keeton KS.A 9-year-old Arabian mare was evaluated for a 7-day history of malaise. Results of a CBC included a leukocyte concentration within the reference interval (8.4 x 10(3)/microL, reference interval 6.0-14.0 x 10(3)/microL) with an apparent degenerative left shift (segmented neutrophils 1.2 x 10(3)/microL, reference interval 2.5-7.5 x 10(3)/microL; hyposegmented neutrophils 1.8 x 10(3)/microL, reference interval 0.0-0.2 x 10(3)/microL). Serum clinical chemistry results included increased aspartate transaminase, alkaline phosphatase, and gamma-glutamyltransferase activities. A presumptive diagnosis ...
Hansen PJ, Hoggard MP, Rathwell AC.The ability of stallion seminal plasma to modify phagocytosis of spermatozoa and Streptococcus zooepidemicus was examined. Phagocytosis was monitored indirectly as the H2O2 produced by peripheral blood leukocytes after addition of spermatozoa or bacteria. Hydrogen peroxide production after addition of ejaculated spermatozoa was greater (P less than 0.01) than after addition of epididymal sperm. Furthermore, pre-incubation of epididymal sperm with 6.25-50% seminal plasma caused a dose-dependent increase in subsequent H2O2 production by leukocytes (P less than 0.05). In addition, equine serum wa...
