Microscopy in equine research involves the use of various microscopic techniques to study the cellular and subcellular structures of horses. This field encompasses the examination of tissues, cells, and microorganisms to gain insights into equine health, disease mechanisms, and physiological processes. Techniques such as light microscopy, electron microscopy, and fluorescence microscopy are employed to observe and analyze samples at high magnification, providing detailed information on morphology and pathology. Microscopy aids in the diagnosis of diseases, identification of pathogens, and evaluation of cellular responses to treatments. This page compiles peer-reviewed research studies and scholarly articles that explore the applications, methodologies, and findings of microscopy in the study of equine biology and medicine.
Buzzell GR, Tariq S, Traversa D, Schuster R.The infective larva of the spirurid nematode Habronema muscae, a parasite of houseflies, was measured and specimens fixed in Karnovsky's fluid were examined by scanning electron microscopy. The oral opening contains six teeth and is surrounded by large bilobed dorsal and ventral lips and smaller lateral lips. A pair of amphids lie behind the lateral lips. There are two rows of four cephalic papillae. The body is deeply ridged, both transversely and longitudinally. The caudal end of the worm is studded by small papillae. The position of the anal opening is somewhat ambiguous. These larval morph...
Rubio-Martínez LM, Cruz AM, Inglis D, Hurtig MB.To determine the anisotropic characteristics of the microarchitecture of the subchondral bone (SCB) plate and trabecular bone (TBB) of the distopalmar aspect of the metacarpal condyles in horses with different stages of SCB disease. Methods: 12 third metacarpal bone pairs from racing Thoroughbreds euthanized for diverse reasons. Methods: Both metacarpi were collected from horses with SCB changes that were mild (sclerosis and focal radiolucencies; n=6) or severe (multifocal radiolucencies and articular surface defects; 6). Sample blocks of SCB plate and TBB were collected from the distopalmar a...
Kulin RM, Jiang F, Vecchio KS.Although clinical bone fractures occur predominantly under impact loading (as occurs during sporting accidents, falls, high-speed impacts or other catastrophic events), experimentally validated studies on the dynamic fracture behavior of bone, at the loading rates associated with such events, remain limited. In this study, a series of tests were performed on femoral specimens obtained post-mortem from equine donors ranging in age from 6 months to 28 years. Fracture toughness and compressive tests were performed under both quasi-static and dynamic loading conditions in order to determine the ef...
Vukojević V, Bowen AM, Wilhelm K, Ming Y, Ce Z, Schleucher J, Hore PJ, Terenius L, Morozova-Roche LA.Recent evidence supports the idea that early aggregates, protein, and lipoprotein oligomers but not large aggregates like fibrils that are formed at late stages of the aggregation process are responsible for cytotoxicity. Oligomers can interact with the cellular plasma membrane affecting its structure and/or dynamics or may be taken up by the cells. In either case, disparate cascades of molecular interactions are activated in the attempt to counteract the disturbance induced by the oligomers. If unsuccessful, cell death follows. Here, we study the molecular and cellular mechanisms underlying P...
Fortier LA, Potter HG, Rickey EJ, Schnabel LV, Foo LF, Chong LR, Stokol T, Cheetham J, Nixon AJ.The purpose of this study was to compare the outcomes of treatment with bone marrow aspirate concentrate, a simple, one-step, autogenous, and arthroscopically applicable method, with the outcomes of microfracture with regard to the repair of full-thickness cartilage defects in an equine model. Methods: Extensive (15-mm-diameter) full-thickness cartilage defects were created on the lateral trochlear ridge of the femur in twelve horses. Bone marrow was aspirated from the sternum and centrifuged to generate the bone marrow concentrate. The defects were treated with bone marrow concentrate and mic...
Skedros JG, Kiser CJ, Mendenhall SD.Using circularly polarized light microscopy,we described a weighted-scoring method for quantifying regional distributions of six secondary osteon morphotypes(Skedros et al.: Bone 44 (2009) 392-403). This osteon morphotype score (MTS) strongly correlated with "tension" and "compression" cortices produced by habitual bending. In the present study, we hypothesized that the osteon MTS is superior to a relatively simpler method based on the percent prevalence (PP) of these osteon morphotypes. This was tested in proximal femoral diaphyses of adult chimpanzees and habitually bent bones: calcanei from...
Schöniger S, Valentine BA, Fernandez CJ, Summers BA.Schwannomas are uncommonly recognized in horses. This study describes cutaneous schwannomas in 22 horses aged 8 to 25 years: 12 male, 7 female, and 3 of unknown sex. The horses had solitary cutaneous masses: 9 on the head, 3 on the neck, and the others on the shoulder, hip, thorax, abdomen, rump, extremities, or tail. The location of 1 tumor was unknown. The dermal tumors were well demarcated and expansile. Twelve had a multinodular pattern, whereas 10 formed a single nodule. Antoni A areas were observed in all tumors, and 10 tumors contained Antoni B areas. In Antoni A areas, the densely pack...
Lee HY, Kopesky PW, Plaas A, Sandy J, Kisiday J, Frisbie D, Grodzinsky AJ, Ortiz C.To quantify the structural characteristics and nanomechanical properties of aggrecan produced by adult bone marrow stromal cells (BMSCs) in peptide hydrogel scaffolds and compare to aggrecan from adult articular cartilage. Methods: Adult equine BMSCs were encapsulated in 3D-peptide hydrogels and cultured for 21 days with TGF-β1 to induce chondrogenic differentiation. BMSC-aggrecan was extracted and compared with aggrecan from age-matched adult equine articular cartilage. Single molecules of aggrecan were visualized by atomic force microscopy-based imaging and aggrecan nanomechanical stiffness...
Kleinheinz J, Jung S, Wermker K, Fischer C, Joos U.Current approaches in bone regeneration combine osteoconductive scaffolds with bioactive cytokines like BMP or VEGF. The idea of our in-vitro trial was to apply VEGF165 in gradient concentrations to an equine collagen carrier and to study pharmacological and morphological characteristics of the complex in a circulation model. Methods: Release kinetics of VEGF165 complexed in different quantities in a collagen matrix were determined in a circulation model by quantifying protein concentration with ELISA over a period of 5 days. The structural changes of the collagen matrix were assessed with lig...
Anthenill LA, Gardner IA, Pool RR, Garcia TC, Stover SM.To compare macrostructural and microstructural features of proximal sesamoid bones (PSBs) from horses with and without PSB midbody fracture to gain insight into the pathogenesis of PSB fracture. Methods: PSBs from 16 Thoroughbred racehorses (8 with and 8 without a PSB midbody fracture). Methods: Parasagittal sections of fractured and contralateral intact PSBs from horses with a PSB fracture and an intact PSB from age- and sex-matched control horses without a PSB fracture were evaluated for visual, radiographic, microradiographic, histologic, and his-tomorphometric differences in bone porosity,...
Loesch A, Mayhew TM, Tang H, Ladd FV, Ladd AA, de Melo MP, da Silva AA, Coppi AA.The superior cervical ganglion (SCG) in mammals varies in structure according to developmental age, body size, gender, lateral asymmetry, the size and nuclear content of neurons and the complexity and synaptic coverage of their dendritic trees. In small and medium-sized mammals, neuron number and size increase from birth to adulthood and, in phylogenetic studies, vary with body size. However, recent studies on larger animals suggest that body weight does not, in general, accurately predict neuron number. We have applied design-based stereological tools at the light-microscopic level to assess ...
Doube M, Firth EC, Boyde A, Bushby AJ.Condylar fracture of the third metacarpal bone (Mc3) is the commonest cause of racetrack fatality in Thoroughbred horses. Linear defects involving hyaline articular cartilage, articular calcified cartilage (ACC) and subchondral bone (SCB) have been associated with the fracture initiation site, which lies in the sagittal grooves of the Mc3 condyle. We discovered areas of thickened and abnormally-mineralised ACC in the sagittal grooves of several normal 18-month-old horses, at the same site that linear defects and condylar fracture occur in older Thoroughbreds and questioned whether this tissue ...
Roelfstra L, Vlimant M, Betschart B, Pfister K, Diehl PA.A morphological study of the midgut and salivary glands of second and third instars of Gasterophilus intestinalis (De Geer) (Diptera: Oestridae) was conducted by light, scanning and transmission electron microscopy. The midgut is anteriorly delimited by a proventriculus, without caeca, and is composed of posterior foregut and anterior midgut tissue from which a double-layered peritrophic matrix is produced. The midgut can be divided into anterior, median and posterior regions on the basis of the structural and physiological variations of the columnar cells which occur along its length. Two oth...
Veronesi MC, Villani M, Wilsher S, Contri A, Carluccio A.The aim of the study was to compare horse and donkey placentae using stereological techniques. Term placentae were collected at spontaneous foaling from seven Thoroughbred mares, seven pony mares, and six jenny donkeys. Maternal and foal weights were recorded and the mass, volume, and gross area of each allantochorion was also recorded. Ten random biopsies were recovered and processed for light microscopy from which the surface density of the microcotyledons (S(v)) and the total microscopic area of fetomaternal contact were calculated stereologically. Gestation length was longer in the donkeys...
Vasconcelos AB, Santana MA, Santos AM, Santoro MM, Lagares MA.Microscopy has been used in the routine evaluation of sperm metabolism. Nevertheless, it has limited capacity to preview male fertility. As calorimetry may be used to evaluate directly the metabolic activity of a biological system, the aim of this study was to use microcalorimetry as an additive method for sperm metabolism evaluation of cooled equine semen. Two ejaculates of four stallions were collected and motility, viability (eosin 3%) and membrane functional integrity (hyposmotic swelling test) of spermatozoa were evaluated. Sperm samples were processed following different protocols and th...
Turowska A, Pajak B, Godlewski MM, Dzieciatkowski T, Chmielewska A, Tucholska A, Banbura M.Viruses can reorganize the cytoskeleton and restructure the host cell transport machinery. During infection viruses use different cellular cues and signals to enlist the cytoskeleton for their mission. However, each virus specifically affects the cytoskeleton structure. Thus, the aim of our study was to investigate the cytoskeletal changes in homologous equine dermal (ED) and heterologous Vero cell lines infected with either equine herpesvirus 1 (EHV-1) strain Rac-H or Jan-E. We found that Rac-H strain disrupted actin fibers and reduced F-actin level in ED cells, whereas the virus did not infl...
Nielsen SB, Wilhelm K, Vad B, Schleucher J, Morozova-Roche LA, Otzen D.The normal function of equine lysozyme (EL) is the hydrolysis of peptidoglycan residues of bacterial cell walls. EL is closely related to alpha-lactalbumins with respect to sequence and structure and further possesses the calcium binding site of alpha-lactalbumins. Recently, EL multimeric complexes with oleic acids (ELOAs) were shown to possess tinctorial and morphological properties, similar to amyloidal aggregates, and to be cytotoxic. ELOA's interactions with phospholipid membranes appear to be central to its biological action, similar to human alpha-lactalbumin made lethal to tumor cells. ...
Matsuda K, Tabata S, Kawamura Y, Kurosawa T, Yoshie N, Taniyama H.Ectopic bone formation in the left atrioventricular valves and cardiac fibroskeleton, with systemic circulatory disturbance, is reported in a 4-year-old crossbred heavy horse. Microscopically, there was fibrosis, chondral metaplasia and mature bone, with bone marrow within the left atrioventricular cusps and in the annuli of the aortic and right atrioventricular valves.
Unger-Torroledo L, Straub R, Lehmann AD, Graber F, Stahl C, Frey J, Gerber V, Hoppeler H, Baum O.The lethal toxin of Clostridium sordellii (TcsL) evokes severe, mostly fatal disease patterns like toxic shock syndrome in humans and animals. Since this large clostridial toxin-induced severe muscle damaging when injected intramuscularly into mice, we hypothesized that TcsL is also associated with equine atypical myopathy (EAM), a fatal myodystrophy of hitherto unknown etiology. Transmission electron microscopy revealed skeletal and heart muscles of EAM-affected horses to undergo degeneration ultrastructurally similar to the damage found in TcsL-treated mice. Performing immunohistochemistry, ...
Kalinová Z, Cisláková L, Halánová M.Ehrlichiosis and anaplasmosis are zoonoses caused by bacteria from the family Anaplasmataceae, including human and animal pathogens. The human pathogens are Ehrlichia chaffeensis, the causative agent of human monocytic ehrlichiosis (HME), Anaplasma phagocytophilum, the pathogen causing human granulocytic anaplasmosis (HGA), E. ewingii and Neorickettsia sennetsu, granulocytotropic and monocytotropic Ehrlichia species, respectively. Ehrlichia spp. are small, gram-negative, obligate intracellular bacteria. They replicate in the cytoplasmic vacuoles of host cells, especially granulocytes and monoc...
Walter I, Tschulenk W, Budik S, Aurich C.The present study gives a detailed ultrastructural description of equine conceptuses at Day 14 (n = 2) and Day 16 (n = 3) after ovulation. Whereas on Day 14 only primitive structures were seen, on Day 16 neurulation and formation of mesodermal somites had taken place. The ectoderm of the embryo itself and the surrounding trophoblast ectodermal cells were characterised by specific cell surface differentiations. At the embryonic ectodermal cell surface (14 and 16 days) remarkable protruded and fused cytoplasmic projections were seen, typically associated with macropinocytotic events involved in ...
Ekmann A, Rigdal ML, Gröndahl G.Microscopy is usually used to obtain manual total and differential cell counts in equine synovial fluid. A faster, more precise method is desirable. Objective: The objectives were to compare an automated impedance method with a manual method for obtaining total and differential cell counts in equine synovial fluid and to evaluate the effect of pretreatment with hyaluronidase on automated results. Methods: Synovial fluid samples (n=48) were collected into EDTA and analyzed within 48 hours. Automated total and differential cell counts were evaluated using a Medonic CA620-VET hematology analyzer ...
Junginger J, Rötting A, Staszyk C, Kramer K, Hewicker-Trautwein M.An adult horse was presented with a mass located within the dermis and subcutis of the right forelimb. At post-mortem examination there was a marked increase in thickness of the subcutis of the right forelimb extending from the fetlock to the pectoral muscles. Microscopically, the dermis and subcutis were diffusely infiltrated by endothelial-like cells forming irregular channels and cavernous spaces. Immunohistochemical examination revealed the endothelial-like cells to express vimentin, factor VIII-related antigen and PROX-1, confirming the identity of the tumour as a lymphangioma. Ultrastruc...
Apprich V, Spergser J, Rosengarten R, Hinterhofer C, Stanek C.Horn samples were taken from the hooves of eight horses with clinical signs of equine onychomycosis in at least one hoof capsule. None of the horses had a documented mycological history. The predominant alterations of the horn capsules were sand cracks, white line disease, brittleness (especially around the nail holes), parakeratosis and bruising. The horn samples were stored in sterile tubes for transportation and transferred onto Sabouraud Dextrose Agar and dermatophyte test agar for mycological examination within 6 h. Fungal cultures were incubated for 30 days at room temperature. Funga...
Nourian AR, Asplin KE, McGowan CM, Sillence MN, Pollitt CC.Anatomical changes in the hoof lamellar tissue induced by prolonged hyperinsulinaemia have not been described previously. Analysis of the induced lesions may promote understanding of hyperinsulinaemic laminitis pathogenesis and produce clinical benefit. Objective: To use light and transmission electron microscopy (TEM) to document hoof lamellar lesions in ponies clinically lame after prolonged hyperinsulinaemia. Methods: Nine clinically normal, mature ponies were allocated randomly to either a treatment group (n = 5) or control group (n = 4). The treatment group received insulin via a modified...
Ledbetter EC, Scarlett JM.To describe morphologic features, pachymetry and endothelial cell density of the normal equine cornea and limbus by in vivo confocal microscopy. Methods: Ten horses without ocular disease. Methods: The central and peripheral corneas were examined with a modified Heidelberg Retina Tomograph II and Rostock Cornea Module using a combination of automated and manual image acquisition modes. Thickness measurements of various corneal layers were performed and endothelial cell density determined. Results: Images of the constituent cellular and noncellular elements of the corneal epithelium, stroma, en...
Schwab UE, Fulcher ML, Randell SH, Flaminio MJ, Russell DG.We describe a method for creating differentiated equine bronchial epithelial cell cultures that can be used for in vitro studies including airway disease mechanisms and pathogen-host interactions. Our method is based on the culturing of human tracheobronchial epithelial cells at an air-liquid interface (ALI) in specific serum-free, hormone-supplemented medium. Bronchial epithelial cells are isolated and grown on T-Clear® insert membranes. Within 2 to 3 wk, cells differentiate into ciliated and mucus producing cells as demonstrated by confocal and electron microscopy. Furthermore, the demonstr...
Traversa D, Milillo P, Barnes H, von Samson-Himmelstjerna G, Schurmann S, Demeler J, Otranto D, Lia RP, Perrucci S, Frangipane di Regalbono A....A broad scale study was carried out in 2008 to evaluate the distribution and species-specific occurrence of cyathostomin populations in horse yards from Europe. In total 102 properties and 3123 horses were included in Italy (60 yards and 1646 animals), United Kingdom (22 yards and 737 animals) and Germany (20 yards and 740 animals). Individual faecal samples were examined with a McMaster technique while pooled samples were subjected to the microscopic examination of in vitro cultured larvae and to a Reverse Line Blot (RLB) assay able to molecularly identify the most diffused 13 species of cyat...
Gericota B, Aleman M, Kozikowski TA, Pesavento P, Bollen AW, Madigan JE, Higgins RJ.A 4-year-old Dutch warmblood mare was presented with a 10-month history of ataxia and proprioceptive deficits. Computed tomography defined a large, non-contrast enhancing mass in the left cerebral hemisphere. Necropsy examination revealed a tumour that effaced much of the piriform and temporal lobes. Microscopically the lesion was classified as a grade IV glioblastoma with an oligodendroglial component (GBM-O). The tumour was composed of highly pleomorphic cells organized in different patterns within a fibrillary stroma. There were multiple foci of necrosis. At the periphery of the tumour neop...
Meyer W, Kacza J, Schnapper A, Verspohl J, Hornickel I, Seeger J.Based on cryo-SEM, standard and high resolution TEM, glycoconjugate histochemistry, and microbiological differentiation, the present study demonstrates the colonisation of the epithelium of the equine oesophagus with microorganisms. As particularly apparent using cryo-SEM to illustrate natural conditions, the present microbiota were clearly dominated by bacteria, forming a one-layer system, as attached to and embedded in concentrated mannose/mannan substances covering the outer stratum corneal cells. Bacterial numbers ranged from 5600 to 7200 per mm(2) in the central part of the oesophagus, th...
Mitchell SR, Kempson SA, Dixon PM.The structure of peripheral cementum of mandibular and maxillary cheek teeth was studied by gross observation, light microscopy, and scanning and transmission electron microscopy. Teeth from four horses aged 4, 8, 17, and 30-years-old were examined. The cementum of the occlusal surface was thicker on the buccal aspect when compared with the lingual aspect of mandibular teeth. The cementum of the occlusal surface was thicker on the palatal aspect when compared with the buccal aspect of maxillary teeth. There was no peripheral cementum on the interdental aspects of either mandibular or maxillary...
Staszyk C, Gasse H.The periodontal ligament (PDL) of the hypsodont cheek teeth of the horse meets specific requirements of tooth support: (1) to fix the tooth; (2) to withstand pressure and tension during the laterolateral mastication movement; (3) to restore the original tooth position. These requirements are assumed to be met by the periodontal collagen fiber apparatus and by the vascular system. Distinct fibro-vascular arrangements are described here in order to reveal the structural prerequisites for these distinct functions. Methods: Specimens from nine horses were processed for (1) collagen labeling with f...
Enders AC, Liu IK.The equine blastocyst becomes fixed in position in the uterus on approximately Day 16 of gestation, but allantochorionic villi are not formed until about Day 50. The purpose of this study was to examine evidence that the blastocyst is orientated during this time period, and to determine what morphological features might assist retention of the position of the blastocyst within the uterus. Implantation sites were collected on Days 10-42 of gestation, and the reproductive tracts perfused with fixative for light and electron microscopic examination. The conceptus is found at the bend of a uterine...
Sivaguru M, Eichorst JP, Durgam S, Fried GA, Stewart AA, Stewart MC.Injuries and damage to tendons plague both human and equine athletes. At the site of injuries, various cells congregate to repair and re-structure the collagen. Treatments for collagen injury range from simple procedures such as icing and pharmaceutical treatments to more complex surgeries and the implantation of stem cells. Regardless of the treatment, the level of mechanical stimulation incurred by the recovering tendon is crucial. However, for a given tendon injury, it is not known precisely how much of a load should be applied for an effective recovery. Both too much and too little loading...
Ibrahim S, Hedia M, Taqi MO, Derbala MK, Mahmoud KGM, Ahmed Y, Sosa AS, Saber YHA, Hasanain MH, Nawito MF, Seidel GE.Extracellular vesicles (EVs) are a promising biomarker and play a vital role in cell-cell communication. This study aimed (I) to identify and characterize EVs from low volume uterine lavage (LVL) and serum in mares with endometritis, compared to healthy controls and (II) to measure serum levels of interleukin 6 (IL-6), and prostaglandins (PGF and PGE). Mares were divided into 30 sub-fertile (endometritis) and 20 fertile (controls). Serum and LVL was collected for EV isolation, and determination of serum levels of inflammatory mediators. Characterization and visualization of EVs were done by el...
Slauson DO, Skrabalak DS, Neilsen NR, Zwahlen RD.Equine neutrophils (PMN) were isolated from citrated normal blood by density gradient separation on Ficoll-Hypaque to greater than 96% purity and 98% viability and an average of 3.78 x 10(7) PMN/ml. The agonist C5a des Arg was used in serial dilutions of whole zymosan-activated equine plasma (ZAP) or was partially purified from ZAP by column chromatography. Purified equine PMN exhibited rapid aggregation following incubation with C5a des Arg which was further dependent on the availability of divalent cations, especially Mg++. The microfilament disruptive agent cytochalasin B (5 micrograms/50 m...
Kilgenstein HJ, Schöniger S, Schoon D, Schoon HA.After their retirement from sports, performance mares often show a poor breeding success. The objective of this study was the microscopic evaluation of endometrial biopsies of retired sports mares (n = 189) to search for alterations that may explain subfertility. Mares of this study aged 3-23 years showed endometritis (30%) and endometrosis (77%); mild forms predominated. In regard to those mares biopsied during the breeding season (n = 99), 50% had glandular differentiation disorders, i.e. glandular inactivity (8%) or irregular glandular differentiation (42%). Compared to literature d...
Lombardi F, Palumbo P, Augello FR, Giusti I, Dolo V, Guerrini L, Cifone MG, Giuliani M, Cinque B.The ability of a collagen-based matrix to support cell proliferation, migration, and infiltration has been reported; however, the direct effect of an aqueous collagen suspension on cell cultures has not been studied yet. In this work, the effects of a high-concentration aqueous suspension of a micronized type I equine collagen (EC-I) have been evaluated on a normal mouse fibroblast cell line. Immunofluorescence analysis showed the ability of EC-I to induce a significant increase of type I and III collagen levels, parallel with overexpression of crucial proteins in collagen biosynthesis, matura...
Marshall VL, Secombe C, Nicholls PK.A 6-week-old Warmblood colt foal was presented for investigation of multiple haematoma formation in various locations, poor wound healing and abnormal scar formation. Based on the history and clinical presentation of hyperextensible skin with prolonged skin tenting, the foal was diagnosed with cutaneous asthenia and euthanased because of the poor prognosis. Histopathological and electron microscopic findings were inconclusive. This is the first case report of cutaneous asthenia in a Warmblood horse in Australia. Cutaneous asthenia is reviewed with particular reference to hereditary equine regi...
Wang X, Thomas CD, Clement JG, Das R, Davies H, Fernandez JW.In this study, the development of a mechanostatistical model of three-dimensional cortical bone remodelling informed with in vivo equine data is presented. The equine model was chosen as it is highly translational to the human condition due to similar Haversian systems, availability of in vivo bone strain and biomarker data, and furthermore, equine models are recommended by the US Federal Drugs Administration for comparative joint research. The model was derived from micro-computed tomography imaged specimens taken from the equine third metacarpal bone, and the Frost-based 'mechanostat' was in...
Perrett J, Harris IT, Maddock C, Farnworth M, Pyatt AZ, Sumner RN.Over the past five decades, there has been increasing evidence to indicate global declines in human semen quality. Parallel adverse trends measured in male animals indicate a potential environmental aetiology. This study evaluated the progressive motility (PM) of stallion ejaculate through a systematic review and meta-analysis. A total of 696 estimates of equine PM from 280 studies, which collected semen samples between the years 1990 and 2018, were collated for meta-analysis. The method of motility analysis, breed, season of collection, and geographical location were extracted. Simple linear ...
Leemans B, Bromfield EG, Stout TAE, Vos M, Van Der Ham H, Van Beek R, Van Soom A, Gadella BM, Henning H.We describe the development of two methods for obtaining confluent monolayers of polarized, differentiated equine oviduct epithelial cells (EOEC) in Transwell inserts and microfluidic chips. EOECs from the ampulla were isolated post-mortem and seeded either (1) directly onto a microporous membrane as differentiated EOECs (direct seeding protocol) or (2) first cultured to a confluent de-differentiated monolayer in conventional wells, then trypsinized and seeded onto a microporous membrane (re-differentiation protocol). Maintenance or induction of EOEC differentiation in these systems was achiev...
Dietze K, Slosarek I, Fuhrmann-Selter T, Hopperdietzel C, Plendl J, Kaessmeyer S.Arterial or venous thromboses are frequent clinical complications with the risk of fatal progression. Recent studies suggest the disruption of angiogenesis in the course of thrombus resolution as the underlying pathomechanism. Very similar to the situation in human patients, equine vessels have been described to be particularly susceptible to thrombosis. In contrast to humans, equine donors are readily available to obtain organs and tissues for isolation of endothelial cells. Objective of this study was to isolate equine endothelial cells and develop an angiogenesis assay from primary cultures...
Eagle RC, Font RL, Swerczek TW.An 18-month-old Standardbred filly had a large intraocular tumor involving the optic nerve. The tumor was a malignant medulloepithelioma, a rare intraocular neoplasm derived from the primitive medullary epithelium. By light microscopy the tumor had cords and lobules of primitive neuroepithelial cells that formed clefts and true rosettes. Electron microscopy of the rosettes showed a girdle of zonulae adherentes joining the apices of the cells as well as several basal bodies. This is the sixth report of equine intraocular medulloepithelioma, and, to the best of our knowledge, the first intraocul...
Dykgraaf S, Firth EC, Rogers CW, Kawcak CE.The objective was to determine the effects of early exercise on the articular cartilage and subchondral bone at specific sites of the distal third metacarpal and metatarsal bones of 12 young Thoroughbred horses allowed free choice exercise at pasture. Six of the horses had additional controlled exercise 5 days per week from mean age of 21+/-20 days of age (range: 3-83 days) until 17.1 months of age. Confocal laser scanning microscopy was used to quantify viable and non-viable chondrocytes. Proteoglycan scoring and modified Mankin scoring was performed and subchondral bone mineral density measu...
Wobeser BK, Hill JE, Jackson ML, Kidney BA, Mayer MN, Townsend HG, Allen AL.Equine sarcoids are the most common tumor of horses. Bovine papillomavirus (BPV) has been suggested as the cause of sarcoids. Studies have shown that BPV is present in swabs or biopsies from nonsarcoid-bearing equine skin. Skin biopsies from a variety of different conditions and normal skin from horses with no reported history of sarcoids were examined by polymerase chain reaction (PCR) for the presence of BPV, which was found in all different types of skin conditions as well as normal skin. Forty-one out of 86 skin biopsies from horses without sarcoids were found to contain BPV DNA. Laser mic...
Gader AG, Ghumlas AK, Hussain MF, Haidari AA, White JG.Previous studies indicated that the camel has a very active haemostatic mechanism with a short bleeding time and thrombocytosis. However, platelet function, when tested by agonist-induced aggregation and PFA 100 closure time, showed marked inhibition compared to humans. Since camels are also far more resistant to long exposure to excessive heat and high body temperature than humans, it seemed worthwhile to explore fundamental morphological differences between human and camel platelets and those from other species. The present study has examined the ultrastructure of camel platelets and compare...
Sharma R, Barakzai SZ, Taylor SE, Donadeu FX.Despite the high prevalence of skin conditions in the horse, there is a dearth of literature on the culture and biology of equine skin cells, and this is partially attributable to the lack of suitable in vitro skin models. The objective of this study was to develop a three-dimensional (3D) culture system that would support the proliferation and differentiation of equine keratinocytes, similar to that observed in natural epidermis. Cell monolayers were obtained from explants of equine skin and serially passaged as highly pure keratinocyte populations (> 95% of cells), based on their express...
Minelli A, Moroni M, Martínez E, Mezzasoma I, Ronquist G.Equine seminal plasma was shown to contain membrane vesicles that are similar to the well characterized prostasomes in human seminal plasma. Determination of nucleoside and nucleotide concentrations of these particles have shown that ATP, ADP and adenosine are the main components of the nucleotidic pool. 5' nucleotidase, endopeptidase and dipeptidyl peptidase i.v. activities have been found on the surface of the particles. The interaction between these prostasome-like vesicles and spermatozoa was demonstrated by electron micrograph scans which revealed the steps of a fusion-like process leadin...
Dieckmann SM, Hoelzle K, Dieckmann MP, Straube I, Hofmann-Lehmann R, Hoelzle LE.Hemotrophic mycoplasmas (HM) are small, cell wall-less bacteria and infections are known for a wide range of animals. One possible indication of equine HM infection was given in 1978, when a 'haemobartonellosis' outbreak was diagnosed in Nigerian horses by microscopy. However the first molecular proof of HM in horses was not reported until 2010, when a fragment of about 900 bp of the 16S rRNA of the equine HM was obtained. This sequence was used for the development of a SYBR green I real-time PCR assay specific for equine HM. The lower detection limit of the PCR was ten 16S rDNA copy numbers p...
Cluzel C, Blond L, Fontaine P, Olive J, Laverty S.Adult articular cartilage (AC) has a well described multizonal collagen structure. Knowledge of foetal AC organisation and development may provide a prototype for cartilage repair strategies, and improve understanding of structural changes in developmental diseases such as osteochondrosis (OC). The objective of this study was to describe normal development of the spatial architecture of the collagen network of equine AC using 1.5 T magnetic resonance imaging (MRI) and polarised light microscopy (PLM), at sites employed for cartilage repair studies or susceptible to OC. T2-weighted fast-spin e...
Equine piroplasmosis is a disease of equids, caused by tick-borne apicomplexan protozoan pathogens Babesia caballi and Theileria equi, which, according to the World Organisation for Animal Health (OIE), can be diagnosed by enzyme-linked immunosorbent assay (ELISA), immunofluorescent antibody test (IFAT) and polymerase chain reaction (PCR). The present study was conducted to evaluate and compare the assays available for the diagnosis of equine piroplasmosis. Data employed were obtained from 1300 blood samples collected between 2012-2014 from asymptomatic and symptomatic equines (horses and donk...
Majewska AC, Werner A, Sulima P, Luty T.The present study was undertaken to investigate the prevalence of Cryptosporidium infection in horses used for recreational riding as well as in humans. A total of 106 faecal specimens from horses raised in 4 localities of western Poland and 6 stool samples from 3 persons who had constant or sporadic contact with horses were screened microscopically for oocysts using modified Ziehl-Neelsen staining. Enzyme immunoassay (EIA) was additionally used for the detection of coproantigen in human stool samples as well as in 43 randomly selected horse faecal samples. The overall infection rate of horses...
Zweygarth E, Just MC, De Waal DT.By means of an in vitro culture technique, 75 samples of horse blood were examined for Babesia equi, a causative agent of equine piroplasmosis. At the time of culture initiation, 15 samples were microscopically positive for B. equi, and this was subsequently confirmed by culture diagnosis. Sixty samples showed no parasites in Giemsa-stained thin blood smears. However, after the culturing process, parasites were found in blood smears of 36 of these samples. The sensitivity of the in vitro culture method was such that 2.5 microliters (1/40 of the usual volume used for the above-mentioned samples...
Benbarek H, Grülke S, Deby-Dupont G, Deby C, Mathy-Hartert M, Caudron I, Dessy-Doize C, Lamy M, Serteyn D.We studied the interactions of isolated equine neutrophils with endothelial cells in culture, mimicking a situation of acute inflammation. Our main purpose was to demonstrate that the supernatant of activated neutrophils was sufficient to damage endothelial cells. Equine endothelial cells (from carotid arteries) were covered either with increased numbers of equine neutrophils stimulated by phorbol myristate acetate, or with the supernatant collected after an in vitro stimulation of the neutrophils. Cytotoxicity was estimated by the release of preincorporated 51Cr, and by light microscopy obser...
Hitchcock PJ, Brown TM, Corwin D, Hayes SF, Olszewski A, Todd WJ.Examination of recently isolated cultures of three strains of Contagious Equine Metritis Organism grown on specially formulated, serum-free, clear typing medium revealed the presence of numerous colonial opacity variants. These colonies were prepared by a number of fixation and staining techniques and examined by scanning and transmission electron microscopy. Opaque and transparent phenotypes produced copious amounts of extracellular material compared with intermediate-opacity phenotypes which produced little or none. Also unique to intermediate colonies were numerous thin intercellular strand...
Bouillant AM, Becker SA.The successive steps of maturation of seven retroviruses from five species of farm animals and one retrovirus from a mouse were compared in cell cultures. The viruses included three type C oncoviruses, one spumavirus, and three lentiviruses. Although members of the 3 subfamilies shared some gross morphologic features such as budding on plasma membranes, core, and surface projections, differences were noted in the ultrastructural detail of these features. Type C oncoviruses did not show any structural differentiation in identifiable form in the cytoplasm as opposed to characteristic features ob...
Otgonsuren D, Amgalanbaatar T, Narantsatsral S, Enkhtaivan B, Munkhgerel D, Zoljargal M, Davkharbayar B, Myagmarsuren P, Battur B, Battsetseg B....Equine piroplasmosis is a tick-borne disease caused by Theileria equi and Babesia caballi in horses. Because of its impact on horse industry, control of this disease is crucial for endemic countries. The control of equine piroplasmosis may be influenced by the genotypic diversity of T. equi and B. caballi. Mongolia, a country with a thriving livestock industry, is endemic for T. equi and B. caballi. However, nationwide epidemiological surveys have not been conducted to determine the current status of infections and genetic diversity of these two parasite species. Therefore, the objective of th...
Gajadhar AA, Caron JP, Allen JR.Cryptosporidium oocysts were identified by phase contrast microscopy on smears from flotations of greenish-yellow pasty feces obtained from two foals. One foal, a one week old Percheron was recumbent, anorectic and lethargic, believed to be the result of a septicemia of undetermined etiology. Despite therapy and nursing care the animal died. Using light and electron microscopy, numerous stages of Cryptosporidium sp. were seen protruding from the surface of epithelial cells of intestinal villi. The other foal, a six week old Arabian had a mild diarrhea. The diarrhea and passage of oocysts event...
Kayar SR, Hoppeler H, Armstrong RB, Laughlin MH, Lindstedt SL, Jones JH, Conley KR, Taylor CR.The mean minimal capillary transit time was estimated in muscles of various animals using a combination of physiological and morphometric methods. Radioactive microspheres were injected intravascularly in various animals running on a treadmill at maximum oxygen consumption rate (VO2,max) to label blood flow to individual muscles. The muscles were then removed and preserved by standard methods for electron microscopy. The volume density of mitochondria was measured to assess muscle oxidative capacity. Capillary densities in muscle cross-sections, capillary diameters and tortuosities were incorp...
