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Topic:Microscopy
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.
Quantitative analysis of collagen fiber organization in injured tendons using Fourier transform-second harmonic generation imaging. Fourier transform-second harmonic generation (FT-SHG) imaging is used as a technique for evaluating collagenase-induced injury in horse tendons. The differences in collagen fiber organization between normal and injured tendon are quantified. Results indicate that the organization of collagen fibers is regularly oriented in normal tendons and randomly organized in injured tendons. This is further supported through the use of additional metrics, in particular, the number of dark (no/minimal signal) and isotropic (no preferred fiber orientation) regions in the images, and the ratio of forward-to-...
Morphometric and ultrastructural features of the mare oviduct epithelium during oestrus. Morphometric, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) investigations have displayed regional differences in the mare oviductal epithelium. The entire mucosa of the oviduct was lined with a pseudostratified epithelium, which consisted of two distinct cell types, ciliated and non-ciliated. Ciliated cells were predominant in the three different segments of the oviduct and their percentage increased from fimbriae to ampulla and significantly decreased in the isthmus. SEM revealed in the infundibulum finger-like mucosal folds, some of them interconnected, in th...
Evaluation of an arthroscopic approach for transection of the equine collateral sesamoidean ligament. To evaluate: (1) an arthroscopic technique for transection of the collateral sesamoidean ligament (CSL); and (2) the healing response using magnetic resonance (MR) and microscopic examination. Methods: Experimental study. Methods: Adult horses (n=6). Methods: Six sound horses with normal front foot radiographic and MR examinations were used. Lameness examination was performed before surgery and monthly for 12 months. Front foot radiography was performed at 180 and 360 days after surgery. Front foot MR was performed before, and at 7, 90, 180, and 360 days after surgery. Arthroscopic CSL desmoto...
A polarized light microscopy method for accurate and reliable grading of collagen organization in cartilage repair. Collagen organization, a feature that is critical for cartilage load bearing and durability, is not adequately assessed in cartilage repair tissue by present histological scoring systems. Our objectives were to develop a new polarized light microscopy (PLM) score for collagen organization and to test its reliability. Methods: This PLM score uses an ordinal scale of 0-5 to rate the extent that collagen network organization resembles that of young adult hyaline articular cartilage (score of 5) vs a totally disorganized tissue (score of 0). Inter-reader reliability was assessed using Intraclass C...
Morphology of the infective larval stage of the equid parasite Habronema muscae (Spirurida: Habronematidae), from houseflies (Musca domestica). 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...
Analysis of the subchondral microarchitecture of the distopalmar aspect of the third metacarpal bone in racing Thoroughbreds. 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...
Effects of age and loading rate on equine cortical bone failure. 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...
Lipoprotein complex of equine lysozyme with oleic acid (ELOA) interactions with the plasma membrane of live cells. 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...
Concentrated bone marrow aspirate improves full-thickness cartilage repair compared with microfracture in the equine model. 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...
A weighted osteon morphotype score outperforms regional osteon percent prevalence calculations for interpreting cortical bone adaptation. 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...
Cutaneous schwannomas in 22 horses. 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...
Adult bone marrow stromal cell-based tissue-engineered aggrecan exhibits ultrastructure and nanomechanical properties superior to native cartilage. 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...
Release kinetics of VEGF165 from a collagen matrix and structural matrix changes in a circulation model. 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...
Comparison of macrostructural and microstructural bone features in Thoroughbred racehorses with and without midbody fracture of the proximal sesamoid bone. 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,...
Stereological and allometric studies on neurons and axo-dendritic synapses in the superior cervical ganglia of rats, capybaras and horses. 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 ...
Combined nanoindentation testing and scanning electron microscopy of bone and articular calcified cartilage in an equine fracture predilection site. 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 ...
Light and electron microscopy studies of the midgut and salivary glands of second and third instars of the horse stomach bot, Gasterophilus intestinalis. 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...
A comparative stereological study of the term placenta in the donkey, pony and Thoroughbred. 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...
Metabolic evaluation of cooled equine spermatozoa. 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...
Opposite effects of two different strains of equine herpesvirus 1 infection on cytoskeleton composition in equine dermal ED and African green monkey kidney Vero cell lines: application of scanning cytometry and confocal-microscopy-based image analysis in a quantitative study. 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...
The interaction of equine lysozyme:oleic acid complexes with lipid membranes suggests a cargo off-loading mechanism. 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. ...
Ectopic ossification with haematopoietic bone marrow in the heart valves of a crossbred heavy horse. 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.
Lethal toxin of Clostridium sordellii is associated with fatal equine atypical myopathy. 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, ...
[Ehrlichiosis/Anaplasmosis]. 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...
Transmission electron microscopy (TEM) of equine conceptuses at 14 and 16 days of gestation. 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 ...
Automated counting of nucleated cells in equine synovial fluid without and with hyaluronidase pretreatment. 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 ...
Identification of equine cutaneous lymphangioma by application of a lymphatic endothelial cell marker. 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...
Scanning electron microscopy and fungal culture of hoof horn from horses suffering from onychomycosis. 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...
Equine laminitis: ultrastructural lesions detected in ponies following hyperinsulinaemia. 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...
In vivo confocal microscopy of the normal equine cornea and limbus. 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...
