Ocular dimensions, corneal thickness, and corneal curvature in quarter horses with hereditary equine regional dermal asthenia

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Lipid structural information from a single equine embryo by MALDI-MS

O objetivo deste trabalho foi relatar o potencial da técnica de MALDI-MS para caracterizar espécies de lipídios presentes em um único embrião equino e estudar algumas estruturas lipídicas detectadas por dissociação induzida por colisão (CID). No espectro de modo íon positivo, pudemos observar espécies, principalmente, protonadas e sodiadas de esfingomielinas (SM), fosfatidileolinas (PC) e triacilgliceróis (TAG). No modo negativo, observamos fosfatidiletanolaminas (PE) e fosfatidilinositos (PI). Espectros de íons de lípidos com maior intensidade foram utilizados para demonstrar o potencial da informação estrutural por MALDI-MS/MS. O espectro no modo positivo de m/z (massa sobre carga) 760,6 (atribuída como PC34:1) apresentou características de fragmentos PC de m/z 184,1 (denominada cabeça polar de colina), além de perda neutral (NL) de m/z 183 (fosforilcolina). Para o íon de m/z 766,6 (atribuída como PE38:5), observou-se a NL de 140, característica do PE. Para o íon de m/z 808,7 (38,5 atribuído como PC), além do fragmento m/z 184,1 na NL de 183, foi possível observar a perda de trimetilamina (íon de m/z 749,6) e o ciclofosfano (íon de m/z 147,0). Finalmente, para o modo de íon negativo, foram isolados e fragmentados o íon de m/z 863,6 que foi atribuído como PI36:1, devido à presença de m/z 153 (fosfato de glicerol – H2O-H ), 223 (inositol fosfo - 2H2O-H) , 241 (fosfoinositol – H2O-H), 281 (ácido oleico) e 581,3 (lisofosfoinositol – H2O+H). Concluímos que a MALDI - MS permite a detecção de uma ampla gama de espécies de PC, SM, PE, PI e TAG lipídicas, bem como a caracterização rápida e confiante de estruturas lipídicas a partir de um único embrião equino.

Use of power doppler for the assessment of vascularization in equine tendinitis treated with mesenchymal stem cells

Tendinous lesions are very common in athlete horses. The process of tendon healing is slow and the quality of the new tissue is often inferior to the original, leading in many cases to recurrence of the lesion. One of the main reasons for the limited healing capacity of tendons is its poor vascularization. At present, cell therapy is used in equine practice for the treatment of several disorders including tendinitis, desmitis and joint disease. However, there is little information regarding the mechanisms of action of these cells during tissue repair. It is known that Mesenchymal Stem Cells (MSCs) release several growth factors at the site of implantation, some of which promote angiogenesis. Comparison of blood flow using power Doppler ultrasonography was performed after the induction superficial digital flexor tendon tendinitis and implantation of adipose tissue-derived MSCs in order to analyze the effect of cell therapy on tendon neovascularization. For quantification of blood vessel histopathological examinations were conducted. Increased blood flow and number of vessels was observed in treated tendons up to 30 days after cell implantation, suggesting promotion of angiogenesis by the cell therapy.

Microbiologic characterization of equine mastitis

Mastitis occurrence in mares is low if compared to other livestock species. The microorganisms often isolated and detected in milk and mammary gland secretions of mares are Streptococcus beta-haemolytica, Staphylococcus spp., Pseudomonas aeruginosa, Actinobacillus spp., and enterobacter. The present experiment was designed to evaluate the main microorganisms present in the milk of healthy mares and having a mammary infection. One hundred and ten mammary glands from 55 lactating mares were analyzed, ranging from 15 to 150 d post-partum. The mastitis diagnostic was performed through analysis of the milk via the screened test of the mug with dark background (Tamis), mammary gland inflammation and/or systemic signs. The subclinical mammary gland infection was characterized via the California Mastitis Test (CMT). From the 55 lactating mares, 2 (3.64%) had clinical mastitis. Following the CMT, the mares presented: 13 (23.60%), 7 (12.72%), and 12 (21.88%) scores from 1+, 2+, and 3+, respectively. From the 110 mamary glands were analysed, in 47 (85.45%) of these samples strains of microorganisms were isolated. In summary, results from our experiment suggest a low occurrence of clinical mastitis in lactating mares.

Neural differentiation from equine inner cell mass cells in vitro

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Protein content of equine seminal plasma and sperm plasma membrane in subfertile stallions

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Development of a cryopreservation protocol for equine mesenchymal stem cells obtained from umbilical cord

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Dinâmica de corpos lúteos em protocolos com redução da dose de FSH na superovulação em ovelhas

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Seroprevalence of equine influenza virus in northeast and southern Mexico

EQUINE influenza A virus (EIV) is a highly infectious respiratory pathogen of horses (Hannant and Mumford 1996, Palese and Shaw 2007). The illness is characterized by an abrupt onset of fever, depression, coughing and nasal discharge, and is often complicated by secondary bacterial infections that can lead to pneumonia and death. Two subtypes of EIV, H3N8 and H7N7, have been isolated. The H7N7 subtype was first isolated from a horse in Czechoslovakia in 1956 (Prague/56), and the H3N8 subtype was first isolated from a horse in Miami in 1963 (Sovinova and others 1958, Waddell and others 1963). The last confirmed outbreak of H7N7 occurred in 1979, and this subtype is now considered to be either extinct or circulating at low levels in a few geographical areas (Ismail and others 1990, Webster 1993, Singh 1994, Madic and others 1996, van Maanen and Cullinane 2002). The H3N8 subtype is a common cause of disease in horses worldwide, particularly in areas where vaccination is not routinely performed (Paillot and others 2006).

Topical Treatment Using Amphotericin B and DMSO for an Atypically Located Equine Cutaneous Pythiosis

Background: Cutaneous lesions by Pythium insidiosum infection are commonly observed in horses, especially in those living at flooded environments. Equine pythiosis is characterized by the development of tumoral masses that are frequently located at distal limbs, ventral abdomen, thorax, breast and face. The lesions are usually granulomatous, serosanguineous and ulcerated, most often destroyed by self-mutilation due to the intense pruritus. The proposed treatment includes surgical excision followed by antifungal drugs administration, which can be done systemically or topically. Amphotericin B and dimethyl sulfoxide (DMSO) in association has been successfully used for cutaneous pythiosis topical treatment due to the DMSO property to carry any substance through plasmatic membranes. Case: The present report concerns a 12-year-old mixed breed gelding presenting with self-mutilation of a tumoral mass located at the left flank. The owners reported that the horse had initially presented a small wound that had evolved to a 20-cm in diameter mass in 4 weeks. Tissue samples were collected, processed and stained by the Gomori's methenamine silver (GMS) method. The histopathological analysis revealed Pythium insidiosum hyphae in a granulomatous tissue, especially located at peripheral region, where kunkers were present. Surgical excision of the mass followed by cauterization was indicated as initial treatment, and due to financial reasons, the owners elected only the topical antifungal therapy to control the fungus infection after surgery. Flunixin meglumine was also administrated for five days aiming the control of pain and inflammation. The wound was cleaned with povidone-iodine solution and rinsed with a solution containing, 50 mg, of amphotericin B in 10 mL of sterile water and 10 mL of DMSO. This procedure was carried Out twice a day. The wound healed fast due to an excellent centripetal epithelialization. and the horse was discharged after 64 days showing only 5% of the initial wound area. The owner reported by telephone the complete healing and hair growth 10 days after discharge. Discussion: Despite the atypical location of the tumoral lesion described at the present report, the history and clinical manifestations, especially the intense pruritus, showed similarity with other characteristic reports of equine cutaneous pythiosis. The diagnosis was confirmed by the histopathological examination showing hyphae structures, as described to be evidences of the presence of Pythium insidiosum in the tissue. The surgical procedure was the first step to provide remission of clinical signs, and one day after surgery the pruritus desapeared. After excision of the granulomatous tissue and cauterization, daily topical administration of amphotericin B associated with DMSO was effective in destroying the infectious agent, as observed by the excellent epithelization. A pink granulation tissue grew up providing an ideal surface for epithelial migration and the healing process progressed quickly. Centripetal epithelialization reduced the wound area until 3% of the initial area in 64 days of treatment, when the remaining wound was found almost completely healed and covered with hair. At the present report, the horse presenting pythiosis was only topically treated. The recommended therapy using amphotericin B and DMSO solution was effective, economically viable and low risk, considering that the systemic antifungal therapy usually suggested is expensive and extremely nephrotoxic. The atypical location of the lesion on the left flank shows that any anatomical region can be affected by the fungus, since the conditions for its development were present.

Expression of tlr4, md2 and cd14 in equine blood leukocytes during endotoxin infusion and in intestinal tissues from healthy horses

The expression of tlr4, md2 and cd14 was studied in equine blood leukocytes and in intestinal samples using real time PCR. The stability of three commonly used reference genes, glyceraldehyde-3P-dehydrogenase (GAPDH), hypoxantine ribosyltransferase (HPRT) and succinate dehydrogenase complex subunit A (SDHA), was evaluated using qbase(PLUS). The equine peripheral blood mononuclear cells (eqPBMC) examined were either stimulated in vitro with Phorbol 12-myristate 13-acetate (PMA) and ionomycin or with the CpG oligodeoxynuclotide 2216 (CpG-ODN 2216) or obtained from horses before, during and after infusion of endotoxin. Intestinal tissue from healthy horses was sampled at ileum, right dorsal colon and rectum. Ranking of the three reference genes used for normalisation identified the combination HPRT/SDHA as most suitable both when determined ex vivo in leukocytes obtained from experimentally induced endotoxaemia and in eqPBMC activated in vitro while HPRT/GAPDH were most appropriate for the intestinal samples. The relative amounts of mRNA for TLR4 and MD-2 increased threefold during in vitro activation of the cells with CpG-ODN 2216 but was decreased in cultures stimulated with PMA/ionomycin. A transient elevation in the transcription of tlr4 and md2 was also evident for equine blood leukocytes following endotoxaemia. The levels of mRNA for CD14 on the other hard remained unaffected both during the induction of endotoxaemia and in the in vitro stimulated PBMCs. A low steady expression of TLR4, MD-2 and CD14 mRNA was demonstrated for the intestinal samples with no variation between the intestinal segments analysed. Thus, the foundation for real time PCR based levels of analysis of mRNA for all three components in the equine LPS receptor complex in different intestinal segments was set, making it possible to carry out future expression studies on clinical material. (C) 2012 Elsevier B.V. All rights reserved.

Treatment of Pythiosis in Equine Limbs Using Intravenous Regional Perfusion of Amphotericin B

Objective To evaluate the effects of intravenous regional limb perfusion (IRLP) administration of amphotericin B in horses to treat pythiosis after surgical excision and thermocautery. Study Design Case series. Animals Horses (n = 12) with Pythium insidiosum infection of the distal aspect of the thoracic or pelvic limbs. Methods After surgical excision of granulation tissue and thermocautery, 50 mg amphotericin B was administered by IRLP through a catheter placed in a superficial vein of the affected limb next to the lesion after placing a tourniquet above the injection site. The lesions and locomotor system were evaluated before treatment and at 7, 14, 21, 28, 35, and 60 days. Results Ninety-two percent of horses treated with amphotericin B had complete lesion resolution 35 or 60 days after 1 or 2 IRLP treatments, respectively. IRLP induced limb edema and pain during regional palpation in 42%, and inflammation of the injection site in 33% of horses; however these signs resolved after 14 days. Conclusions IRLP administration of amphotericin B was effective for treating pythiosis in equine limbs, resolving infection with manageable side effects.

Do blood components affect the production of reactive oxygen species (ROS) by equine synovial cells in vitro?

Brossi P.M., Baccarin R.Y.A. & Massoco C.O. 2012 Do blood components affect the production of reactive oxygen species (ROS) by equine synovial cells in vitro? Pesquisa Veterinaria Brasileira 32(12):1355-1360. Departamento de Clinica Medica, Faculdade de Medicina Veterinaria e Zootecnia, Universidade de Sao Paulo, Av. Prof. Dr. Orlando Marques de Paiva 87, Butanta, Sao Paulo, SP 5508-210, Brazil. E-mail: baccarin@ usp.br Blood-derived products are commonly administered to horses and humans to treat many musculoskeletal diseases, due to their potential antioxidant and anti-inflammatory effects. Nevertheless, antioxidant effects have never been shown upon horse synovial fluid cells in vitro. If proved, this could give a new perspective to justify the clinical application of blood-derived products. The aim of the present study was to investigate the antioxidant effects of two blood-derived products - plasma (unconditioned blood product - UBP) and a commercial blood preparation (conditioned blood product - CBP)(4) - upon stimulated equine synovial fluid cells. Healthy tarsocrural joints (60) were tapped to obtain synovial fluid cells; these cells were pooled, processed, stimulated with lipopolysaccharide (LPS) or phorbol 12-myristate 13-acetate (PMA), and evaluated by flow cytometry for the production of reactive oxygen species (ROS). Upon addition of any blood-derived product here used - UBP and CBP - there was a significant decrease in the oxidative burst of synovial fluid cells (P<0.05). There was no difference between UBP and CBP effects. In conclusion, treatment of stimulated equine synovial cells with either UBP or CBP efficiently restored their redox equilibrium.