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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.
Development and characterization of an equine skin-equivalent model.
Veterinary dermatology    July 18, 2014   Volume 25, Issue 5 475-e77 doi: 10.1111/vde.12134
Cerrato S, Ramió-Lluch L, Brazís P, Rabanal RM, Fondevila D, Puigdemont A.There is increasing interest in the biological and pathological study of equine skin owing to the high prevalence of cutaneous diseases in horses. However, knowledge of equine skin cell biology and cultures is limited by the low number of in vitro studies in the literature. Objective: The objective of the study was to develop and characterize an in vitro equine skin equivalent. Methods: Cultures of pure equine keratinocytes and dermal fibroblasts were obtained by enzymatic digestion of skin biopsies. Fibroblasts were embedded into type I collagen matrices to obtain dermal scaffolds, the surfac...
The recent European isolate (08P178) of equine arteritis virus causes inflammation but not arteritis in experimentally infected ponies.
Journal of comparative pathology    June 27, 2014   Volume 151, Issue 2-3 238-243 doi: 10.1016/j.jcpa.2014.04.008
Vairo S, Saey V, Bombardi C, Ducatelle R, Nauwynck H.In the last two decades, outbreaks of equine viral arteritis (EVA) have been reported in Europe, but little is known about these European isolates of equine arteritis virus (EAV). EAV European strain (08P178, EU-1 clade) isolated from one of these recent outbreaks is able to cause clinical signs on experimental infection. The aim of the present study was to investigate the microscopical lesions induced by this isolate after experimental infection of ponies. Animals were killed at 3, 7, 14 and 28 days post infection (dpi). At 3 dpi, lesions were essentially restricted to the respiratory tract a...
Characterization of amyloid in equine recurrent uveitis as AA amyloid.
Journal of comparative pathology    June 26, 2014   Volume 151, Issue 2-3 228-233 doi: 10.1016/j.jcpa.2014.04.007
Ostevik L, de Souza GA, Wien TN, Gunnes G, Sørby R.Two horses with chronic uveitis and histological lesions consistent with equine recurrent uveitis (ERU) were examined. Microscopical findings in the ciliary body included deposits of amyloid lining the non-pigmented epithelium, intracytoplasmic, rod-shaped, eosinophilic inclusions and intraepithelial infiltration of T lymphocytes. Ultrastructural examination of the ciliary body of one horse confirmed the presence of abundant extracellular deposits of non-branching fibrils (9-11 nm in diameter) consistent with amyloid. Immunohistochemistry revealed strong positive labelling for AA amyloid and m...
Vitrification of equine oocytes with a polyvinyl alcohol after in vitro maturation with equine growth hormone and insulin-like growth factor-I.
Cryo letters    May 30, 2014   Volume 35, Issue 2 90-94 
Curcio Bda R, Pereira GR, Antunes LI, Boff AN, dos Santos FC, Lucas T, Nogueira CE, Corcini CD, Liu I, Deschamps JC.In vitro fertilization (IVF) procedures are limited by the inability to mature equine oocytes on in vitro methods. Objective: The aim of this study was to evaluate structural integrity of equine oocytes subjected to vitrification with a synthetic polymer (PVA). Methods: The effect of eGH and its relationship with IGF-I on in vitro maturation (IVM) were evaluated. Compact cumulus oocytes complexes (n=122) were cultured in TCM-199 with eGH, IGF-I or eGH+IGF-I for 30h at 38.5C in air with 5 % CO2. Oocytes were fixed after IVM or subjected to the vitrification protocol. Cryopreserved oocytes were ...
Regulation of ACVR1 and ID2 by cell-secreted exosomes during follicle maturation in the mare.
Reproductive biology and endocrinology : RB&E    May 26, 2014   Volume 12 44 doi: 10.1186/1477-7827-12-44
da Silveira JC, Carnevale EM, Winger QA, Bouma GJ.Ovarian follicle growth and maturation requires extensive communication between follicular somatic cells and oocytes. Recently, intercellular cell communication was described involving cell-secreted vesicles called exosomes (50-150 nm), which contain miRNAs and protein, and have been identified in ovarian follicular fluid. The goal of this study was to identify a possible role of exosomes in follicle maturation. Methods: Follicle contents were collected from mares at mid-estrous (~35 mm, before induction of follicular maturation) and pre-ovulatory follicles (30-34 h after induction of folli...
Use of immunohistochemical staining and electron microscopy to aid in diagnosis of soft tissue sarcomas associated with the fetlock joint in two horses. Findley JA, Singer ER, Milner PI, Leeming GH.Soft tissue sarcomas of the equine distal limb associated with joints, sheaths, or bursae have rarely been reported. Accurate diagnosis of these tumors is challenging in both human beings and veterinary species. Immunohistochemical staining and transmission electron microscopy have been used in human beings to reduce misdiagnosis. The current report describes 2 mature horses presenting with lameness and swelling associated with the dorsal aspect of the metacarpo(tarso)phalangeal joint. In both cases, surgical excision was performed with subsequent histological analysis of the masses to determi...
Microstructural changes in cartilage and bone related to repetitive overloading in an equine athlete model.
Journal of anatomy    April 1, 2014   Volume 224, Issue 6 647-658 doi: 10.1111/joa.12177
Turley SM, Thambyah A, Riggs CM, Firth EC, Broom ND.The palmar aspect of the third metacarpal (MC3) condyle of equine athletes is known to be subjected to repetitive overloading that can lead to the accumulation of joint tissue damage, degeneration, and stress fractures, some of which result in catastrophic failure. However, there is still a need to understand at a detailed microstructural level how this damage progresses in the context of the wider joint tissue complex, i.e. the articular surface, the hyaline and calcified cartilage, and the subchondral bone. MC3 bones from non-fractured joints were obtained from the right forelimbs of 16 Thor...
Ultrastructure and growth factor content of equine platelet-rich fibrin gels.
American journal of veterinary research    March 29, 2014   Volume 75, Issue 4 392-401 doi: 10.2460/ajvr.75.4.392
Textor JA, Murphy KC, Leach JK, Tablin F.To compare fiber diameter, pore area, compressive stiffness, gelation properties, and selected growth factor content of platelet-rich fibrin gels (PRFGs) and conventional fibrin gels (FGs). Methods: PRFGs and conventional FGs prepared from the blood of 10 healthy horses. Methods: Autologous fibrinogen was used to form conventional FGs. The PRFGs were formed from autologous platelet-rich plasma of various platelet concentrations (100 × 10³ platelets/μL, 250 × 10³ platelets/μL, 500 × 10³ platelets/μL, and 1,000 × 10³ platelets/μL). All gels contained an identical fibrinogen concentra...
Ultrastructural analysis of healthy synovial fluids in three mammalian species. Matei CI, Boulocher C, Boulé C, Schramme M, Viguier E, Roger T, Berthier Y, Trunfio-Sfarghiu AM, Blanchin MG.A better knowledge of synovial fluid (SF) ultrastructure is required to further understand normal joint lubrication and metabolism. The aim of the present study was to elucidate SF structural features in healthy joints from three mammalian species of different size compared with features in biomimetic SF. High-resolution structural analysis was performed using transmission electron microscopy (TEM) and scanning electron microscopy (SEM) and environmental SEM/wet scanning transmission electron microscopy mode complemented by TEM and SEM cryogenic methods. Laser-scanning confocal microscopy (LCM...
Transferrin receptor expression in serum exosomes as a marker of regenerative anaemia in the horse.
Equine veterinary journal    March 13, 2014   Volume 47, Issue 1 101-106 doi: 10.1111/evj.12235
Rout ED, Webb TL, Laurence HM, Long L, Olver CS.Evaluation of erythrocyte regeneration in horses is challenging, as they do not release reticulocytes into the peripheral blood. This study investigated transferrin receptor 1 (TfR1) expression in exosomes as a noninvasive method of characterising the regenerative response in anaemic horses. Objective: To quantify TfR1 in ultraprecipitate of serum in horses before and after phlebotomy-induced anaemia, and to identify exosomes as the source of TfR1. The hypothesis was that serum exosomal TfR1 expression would increase during a regenerative response. Methods: Experimental model of anaemia. Metho...
Stallion spermatozoa: putative target of estrogens; presence of the estrogen receptors ESR1, ESR2 and identification of the estrogen-membrane receptor GPER.
General and comparative endocrinology    March 4, 2014   Volume 200 35-43 doi: 10.1016/j.ygcen.2014.02.016
Arkoun B, Gautier C, Delalande C, Barrier-Battut I, Guénon I, Goux D, Bouraïma-Lelong H.Among mammals, the stallion produces the largest amount of testicular estrogens. These steroid hormones are produced mainly by Leydig and Sertoli cells in the testis and also in the epididymis. Their role in horse testicular physiology and their ability to act on spermatozoa are still unknown. In order to determine if spermatozoa are targets for estrogens, the presence of estrogen receptors in mature ejaculated spermatozoa has been investigated. The presence of a single isoform of ESR1 (66kDa) and ESR2 (61kDa) was found by Western-blot analysis in samples from seven stallions. Confocal analysi...
The influence of static magnetic fields on canine and equine mesenchymal stem cells derived from adipose tissue.
In vitro cellular & developmental biology. Animal    January 30, 2014   Volume 50, Issue 6 562-571 doi: 10.1007/s11626-013-9730-1
Marędziak M, Marycz K, Smieszek A, Lewandowski D, Toker NY.The aim of this study was to evaluate the proliferation rate and morphological changes of adipose-derived mesenchymal stem cells of canine and equine origin (Eq- and CaAdMSC). Investigated cells were exposed to a static magnetic field (MF) with the intensity of 0.5 T. Proliferation activity of cells was determined with the Alamar Blue assay. Obtained results, normalized in respect to the control culture, showed that EqAdMSC exposed to MF maintained a high proliferation status, whereas proliferation activity of CaAdMSC cultured in the presence of MF was decreased. Estimations of population doub...
Development of coronal cementum in hypsodont horse cheek teeth.
Anatomical record (Hoboken, N.J. : 2007)    January 28, 2014   Volume 297, Issue 4 716-730 doi: 10.1002/ar.22880
Sahara N.The horse is a grazing herbivore whose cheek teeth are hypsodon; that is, they possess long crowns that are completely covered by coronal cement at eruption. For elucidation of the sequential events in the formation of this coronal cementum in the mandibular horse cheek teeth, in the present study the lower 3rd permanent premolar teeth (PM4 ) from 3.5-, 4-, and 5-year-old horses were compared by using radiography, microcomputed tomography (Miro-CT), light microscopy (LM), and scanning electron microscopy (SEM). The present study clearly showed that prior to coronal cementogenesis tartrate-resi...
Acinar cell carcinoma of exocrine pancreas in two horses.
Journal of comparative pathology    January 16, 2014   Volume 150, Issue 4 388-392 doi: 10.1016/j.jcpa.2014.01.003
de Brot S, Junge H, Hilbe M.Two horses were presented with non-specific clinical signs of several weeks' duration and were humanely destroyed due to a poor prognosis. At necropsy examination, both horses had multiple small, white nodules replacing pancreatic tissue and involving the serosal surface of the abdominal cavity, the liver and the lung. Microscopically, neoplastic cells were organized in acini and contained abundant (case 1) or sparse (horse 2) intracytoplasmic zymogen granules. Immunohistochemically, both tumours expressed amylase and pan-cytokeratin, but not insulin or neuron-specific enolase. In case 2, a lo...
In vivo confocal microscopy of corneal microscopic foreign bodies in horses.
Veterinary ophthalmology    January 13, 2014   Volume 17 Suppl 1 69-75 doi: 10.1111/vop.12139
Ledbetter EC, Irby NL, Schaefer DM.To describe in vivo corneal confocal microscopy of horses with microscopic corneal foreign bodies and to correlate findings with clinical, cytological, and histopathologic evaluations of clinical cases and foreign body morphologies observed in vitro with the confocal microscope. Methods: Five horses with microscopic corneal foreign bodies. Methods: Sedated and anesthetized horses were examined with a modified Heidelberg Retina Tomograph II and Rostock Cornea Module. Confocal microscopy images were compared with images from cytologic and histopathologic corneal samples. To establish microscopic...
Dark-field microscopy in the assessment of large colon microperfusion and mucosal injury in naturally occurring surgical disease of the equine large colon.
Equine veterinary journal    January 3, 2014   Volume 46, Issue 6 674-680 doi: 10.1111/evj.12202
Hurcombe SD, Welch BR, Williams JM, Cooper ES, Russell D, Mudge MC.Intraoperative assessment of colonic viability can be challenging and largely subjective. Objective methods are often impractical. Viability is related to re-establishment of colonic perfusion; particularly microvascular perfusion. This study evaluated the utility of dark-field microscopy (DFM) of the colonic serosa as an objective method of assessing microperfusion. Objective: To measure microvascular perfusion indices (MPI) of the pelvic flexure serosa in horses with surgical colonic lesions and correlate these with macroperfusion indices (MaPI) and histomorphometry. Methods: Prospective, cl...
Mosaic arthroplasty of the medial femoral condyle in horses – An experimental study.
Acta veterinaria Hungarica    December 18, 2013   Volume 62, Issue 2 155-168 doi: 10.1556/AVet.2013.059
Bodó G, Vásárhelyi G, Hangody L, Módis L.One Arabian and 5 Hungarian half-bred horses were used to study the macroscopic and microscopic survival of autologous osteochondral grafts in the weight-bearing surface of the medial femoral condyle (MFC). Grafts were harvested from the cranial surface of the medial femoral trochlea (MFT) under arthroscopic control. Three of them were transplanted into the weight-bearing surface of the contralateral MFC using an arthrotomy approach. Three months later this transplantation procedure was repeated on the opposite stifle joints in the same animals, but at that time transplantation was performed a...
Identification of cartilage injury using quantitative multiphoton microscopy.
Osteoarthritis and cartilage    November 1, 2013   Volume 22, Issue 2 355-362 doi: 10.1016/j.joca.2013.10.008
Novakofski KD, Williams RM, Fortier LA, Mohammed HO, Zipfel WR, Bonassar LJ.Cartilage injury can lead to post-traumatic osteoarthritis (PTOA). Immediate post-trauma cellular and structural changes are not widely understood. Furthermore, current cellular-resolution cartilage imaging techniques require sectioning of cartilage and/or use of dyes not suitable for patient imaging. In this study, we used multiphoton microscopy (MPM) data with FDA-approved sodium fluorescein to identify and evaluate the pattern of chondrocyte death after traumatic injury. Methods: Mature equine distal metacarpal or metatarsal osteochondral blocks (OCBs) were injured by 30 MPa compressive loa...
Morphometric evaluation of the spermatogonial stem cell distribution and niche in vertebrates.
Methods in molecular biology (Clifton, N.J.)    August 21, 2013   Volume 1035 35-42 doi: 10.1007/978-1-62703-508-8_4
Campos-Junior PH, Costa GM, de Avelar GF, Segatelli TM, Lacerda SM, Aponte PM, de França LR.Morphometry is a classical quantitative method often used in biology to provide a data basis for functional interpretations/interactions of a particular organ or system. Herein we took advantage of this valuable approach to evaluate the spermatogonial stem cell niche using the horse testis and immunocytochemical localization of GFRA1 [glial cell line-derived neurotrophic factor receptor produced by Sertoli cells)] as an example. Using the NIH ImageJ free software, we describe in detail all the necessary steps to investigate this specific and crucial microenvironment. Based on several recently ...
Foetal and postnatal equine articular cartilage development: magnetic resonance imaging and polarised light microscopy.
European cells & materials    August 9, 2013   Volume 26 33-48 doi: 10.22203/ecm.v026a03
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 oviduct explant culture: a basic model to decipher embryo-maternal communication.
Reproduction, fertility, and development    August 2, 2013   Volume 26, Issue 7 954-966 doi: 10.1071/RD13089
Nelis H, D'Herde K, Goossens K, Vandenberghe L, Leemans B, Forier K, Smits K, Braeckmans K, Peelman L, Van Soom A.Equine embryos remain for 6 days in the oviduct and thus there is a need for an in vitro model to study embryo-oviductal interactions in the horse, since this subtle way of communication is very difficult to analyse in vivo. Until now, no equine oviduct explant culture model has been characterised both morphologically and functionally. Therefore, we established a culture system for equine oviduct explants that maintained epithelial morphology during 6 days of culture, as revealed by light microscopy and transmission electron microscopy. We demonstrated the presence of highly differentiated, ta...
Epidermal-like architecture obtained from equine keratinocytes in three-dimensional cultures.
Journal of tissue engineering and regenerative medicine    July 30, 2013   Volume 10, Issue 8 627-636 doi: 10.1002/term.1788
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...
The involvement of growth hormone in equine oocyte maturation, receptor localization and steroid production by cumulus-oocyte complexes in vitro.
Research in veterinary science    July 26, 2013   Volume 95, Issue 2 667-674 doi: 10.1016/j.rvsc.2013.06.024
Pereira GR, Lorenzo PL, Carneiro GF, Ball BA, Bilodeau-Goeseels S, Kastelic J, Pegoraro LM, Pimentel CA, Esteller-Vico A, Illera JC, Granado GS....The objectives of this study were to evaluate the effects of equine growth hormone (eGH) on nuclear and cytoplasmic maturation of equine oocytes in vitro, steroid production by cumulus cells, and expression and subcellular localization of eGH-receptors (eGH-R) on equine ovarian follicles. Cumulus-oocyte complexes (COCs) were recovered by aspirating follicles <30 mm in diameter from abattoir-derived ovaries. The COCs were morphologically evaluated and randomly allocated to be cultured in either a control maturation medium or supplemented with 400 ng/mL eGH, for 30 h at 38.5°C in air with 5%...
Imaging horse tendons using multimodal 2-photon microscopy.
Methods (San Diego, Calif.)    July 19, 2013   Volume 66, Issue 2 256-267 doi: 10.1016/j.ymeth.2013.07.016
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...
Influence of equine growth hormone, insulin-like growth factor-I and its interaction with gonadotropins on in vitro maturation and cytoskeleton morphology in equine oocytes.
Animal : an international journal of animal bioscience    June 21, 2013   Volume 7, Issue 9 1493-1499 doi: 10.1017/S175173111300116X
Pereira GR, Lorenzo PL, Carneiro GF, Ball BA, Pegoraro LM, Pimentel CA, Liu IK.In horses, successful in vitro fertilization procedures are limited by our inability to consistently mature equine oocytes by in vitro methods. Growth hormone (GH) is an important regulator of female reproduction in mammals, playing an important role in ovarian function, follicular growth and steroidogenesis. The objectives of this research were to investigate: the effects of equine growth hormone (eGH) and insulin-like growth factor-I (IGF-I) on the in vitro maturation (IVM) of equine oocytes, and the effects of eGH in addition to estradiol (E2), gonadotropins (FSH and LH) and fetal calf seru...
Readability and histological biocompatibility of microchip transponders in horses.
Veterinary journal (London, England : 1997)    June 12, 2013   Volume 198, Issue 1 103-108 doi: 10.1016/j.tvjl.2013.04.028
Wulf M, Wohlsein P, Aurich JE, Nees M, Baumgärtner W, Aurich C.Identification of horses by microchip transponder is mandatory within the European Union with only a few exceptions. In this study, the readability of such microchips in 428 horses with three different scanners (A, B and C) and the histological changes at the implantation site in 16 animals were assessed. Identification of microchips differed between scanners (P<0.001), and with 'side of neck' (P<0.001). Scanners A, B and C identified 93.5%, 89.7% and 100% of microchips, respectively, on the 'chip-bearing' side of the neck. From the contralateral side, scanners A, B a...
Functional characterization of detergent-decellularized equine tendon extracellular matrix for tissue engineering applications.
PloS one    May 27, 2013   Volume 8, Issue 5 e64151 doi: 10.1371/journal.pone.0064151
Youngstrom DW, Barrett JG, Jose RR, Kaplan DL.Natural extracellular matrix provides a number of distinct advantages for engineering replacement orthopedic tissue due to its intrinsic functional properties. The goal of this study was to optimize a biologically derived scaffold for tendon tissue engineering using equine flexor digitorum superficialis tendons. We investigated changes in scaffold composition and ultrastructure in response to several mechanical, detergent and enzymatic decellularization protocols using microscopic techniques and a panel of biochemical assays to evaluate total protein, collagen, glycosaminoglycan, and deoxyribo...
Cytotoxic activity of extracts from Hypochaeris radicata.
Toxicon : official journal of the International Society on Toxinology    May 9, 2013   Volume 70 194-203 doi: 10.1016/j.toxicon.2013.04.023
MacKay RJ, Wyer S, Gilmour A, Kongara K, Harding DR, Clark S, Mayhew IG, Thomson CE.Pasture-associated stringhalt is an acquired equine disease characterized by peripheral neuropathy and hyperflexion of the pelvic limbs. The disease occurs most commonly during periods of drought in horses grazing pastures heavily contaminated by Hypochaeris radicata. We hypothesized that stringhalt is caused by neurotoxins elaborated by H. radicata in response to the stress of drought conditions. Supernates were collected from H. radicata that were stressed (or not) by immersion in copper chloride solution, then extracted with ethyl acetate and dried. Dilutions of extracts from stressed (SE) ...
A 3-D airway epithelial cell and macrophage co-culture system to study Rhodococcus equi infection.
Veterinary immunology and immunopathology    May 7, 2013   Volume 154, Issue 1-2 54-61 doi: 10.1016/j.vetimm.2013.05.003
Schwab U, Caldwell S, Matychak MB, Felippe J.We developed a 3-D equine bronchial epithelial cell (BEC) culture that fully differentiates into ciliary beating and mucus producing cells. Using this system, we evaluated how mucus affects the phagocytic activity of macrophages. Adult horse monocyte-derived macrophages were incubated with Rhodococcus equi for 4h either in the mucus layer of in vitro generated airway epithelium or on collagen coated membranes. Using light and electron microscopy, we noted that the number of macrophages with intracellular bacteria, and the number of intracellular bacteria per macrophage were lower in the presen...
A histomorphological analysis of human and non-human femora.
International journal of legal medicine    April 21, 2013   Volume 128, Issue 2 369-377 doi: 10.1007/s00414-013-0854-3
Brits D, Steyn M, L'Abbé EN.Histology is used to describe post-mortem bone alterations, trauma, pathology and age estimation and to separate human and non-human bones. Many scholars are however not familiar with the intricate and variable microstructure of bone, and due to the complex nature of some classification systems, bone histomorphology is often incorrectly described or identified. Little information is available on the histomorphology of non-human bones found in southern Africa, and therefore, the aim of this study was to describe the histomorphology of non-human species commonly found in southern Africa, namely,...
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