Bone marrow derived mesenchymal stem cells (MSCs) can be used to repair articular cartilage defects, these cells should be properly stimulated so that they could differentiate morphologically and hold cellular synthetic features closer to maturely differentiated chondrocytes. It is well known that tissue specific environment plays an important role in cell fate determination. Once improved isolation, proliferation and differentiation protocols have been developed, the likelihood of spontaneous differentiation of MSCs into divergent lineages will be reduced, thus increasing their value for cartilage repair. The purpose of this study was to improve chondrogenic differentiation of equine MSCs using coculture with mature equine articular chondrocytes (ACs), along with the determination of the effect of adding transforming growth factor (TGF) beta1 in the pellet culture system. Following confirmation of multilineage (adipogenic, osteogenic and chondrogenic) differentiation, isolated MSCs, ACs and coculture of both cell types were transferred into pellet culture system in a DMEM-based medium supplemented with or without TGFbetal. Chondrogenic differentiation was evaluated histologically and the relative mRNA expressions of collagen type 1 alpha1 (COL1A1), collagen type 2 alpha1 (COL2A1), aggrecan (ACAN) and SRY-box 9 (SOX9) were estimated by quantitative RT-PCR. Cocultured cells showed diffuse distribution of extracellular matrix (ECM), whereas in chondrocyte pellets it was more localized to central regions. Expression of COL2A1, ACAN and SOX9 genes were higher in cocultured pellets when compared to MSCs and ACs-composed pellets. Addition of TGFbeta1 in chondrogenic differentiating medium did not consistently amplify expression of the above mentioned genes. Differentiation of equine MSCs was enhanced by coculturing in association with mature ACs, improving expression of cartilage-specific genes and producing a more homogeneous production of ECM within the newly formed cocultured cartilage. The use of the coculture system could possibly enhance the capacity of MSC-derived chondrocytes to build up stable articular cartilage-like constructs, which could play an important role in articular cartilage repair and regeneration.

peer reviewed

peer reviewed | Abstract Title: Anoxia / Reoxygenation of equine endothelial cells produce reactive oxygen species: an oxymetric and electron paramagnetic resonance investigation. G. de la Rebière de Pouyade 1,2, A. Mouithys-Mickalad 2, A. Salciccia 1,2, J. Ceusters 2, G. Deby-Dupont 2, D. Serteyn 1,2 1-Department of Clinical Sciences, Large Animal Surgery, Faculty of Veterinary Medicine B41, University of Liège, Sart Tilman, 4000 Liège, Belgium 2-Center for Oxygen Research and Development, Institute of Chemistry B6a, University of Liège, Sart Tilman, 4000 Liège, Belgium Take Home Message: Equine endothelial cells (EC) submitted to anoxia/reoxygenation (A/R) produce reactive oxygen species (ROS). Introduction: Endothelium participates to the regulation of the inflammatory response and ischemia/reperfusion-induced injuries are suspected in laminitis. Herein, we investigated the effect of anoxia/reoxygenation on ROS production by EC. Materials and Methods: EC cultures were obtained from equine carotid. Detached cells (107) were submitted either to 3 cycles of 20min anoxia followed by reoxygenation in presence of POBN (50mM)/ethanol (2%) mixture as spin trap or to 1h of anoxia followed by reoxygenation and immediate addition of 100mM DMPO. Normoxic cells were used as control (n=3). Anoxia and reoxygenation were monitored by high-resolution respirometry and ROS production was evidenced by electron paramagnetic resonance (EPR). Results: After A/R cycles, high intensity EPR spectra, assigned to the POBN/•CH(OH)CH3 adducts were observed, but not in normoxic cells (control). After 1h of anoxia followed by reoxygenation, an enhanced production of DMPO-OH adducts was observed compared to control. Discussion: The EPR spectra were consistent with the trapping of superoxide anion produced by EC. Such a production could derive from either xanthine oxidase activity or mitochondria dysfunction. Production of ROS by EC could be involved in the disruption of the endothelial barrier, the recruitment of neutrophils and participate to the development of laminitis. Clinical relevance: Drugs (fractionated heparin or pentoxifylline) able to protect endothelium may have therapeutic relevance in preventing laminitis. Conclusion: ROS from EC submitted to A/R might play a crucial role in the oxidative aspect of laminitis.

Mastitis is a common illness of dairy cattle and is very costly economically to the dairy farmer. Thus there is a need to develop broad-spectrum therapies that are effective while not leading to unacceptably long antibiotic withdrawal times. The effects of the CH4-inhibitors nitroethane (2 mg/ml), 2-nitro-1-propanol (2 mg/ml), lauric acid (5 mg/ml), the commercial product Lauricidin ® (5 mg/ml), and a finely-ground product of the Hawaiian marine algae, Chaetoceros (10 mg/ml), were compared in pure cultures of S. aureus. Lauricidin® exhibited the most bactericidal acidity against S. aureus. These results suggest potential for treatments with a non antibiotic compound could be effective against mastitis.

Para avaliar o efeito da utilização parenteral das vitaminas C e/ou E associadas ao quelante de cobre, tetratiomolibdato de amônio (TTM), no tratamento de ovinos com intoxicação cumulativa por cobre (ICC), foram analisados o metabolismo oxidativo, através das concentrações sanguíneas de glutationa reduzida (GSH), séricas de malondialdeído (MDA) e ácido úrico, e a habilidade de redução férrica plasmática (HRFP) desses animais. Foram utilizados 24 ovinos machos, da raça Santa Inês, com peso médio de 25 kg e distribuídos em quatro tratamentos: TTM, TTM e vitamina C (TTM+VC), TTM e vitamina E (TTM+VE) e TTM e vitaminas C e E (TTM+VCE). O quadro de intoxicação cúprica provocou intensa formação de radicais livres, com redução da concentração de GSH e aumento do MDA, apesar do aumento na capacidade antioxidante plasmática, decorrente do aumento da concentração de ácido úrico e da HRFP. Não foi observado benefício algum da utilização das vitaminas antioxidantes, isoladamente ou associadas, com o tratamento clássico baseado na administração de TTM, na redução do estresse oxidativo. | The effects of vitamins C and/or E associated to amonium tetrathiomolybdate (TTM) in the treatment of cumulative copper intoxication (ICC) in sheep were analyzed throughout oxidative metabolism. Reduced glutathione (GSH), malondialdehyde (MDA), uric acid and the ferric reducing ability of plasma (FRAP) concentrations were evaluated. Twenty four male sheep, Santa Ines breed, with 25 kg BW was distributed in four treatments: TTM, TTM and vitamin C (TTM+VC), TTM with vitamin E (TTM+VE) and TTM with vitamins C and E (TTM+VCE). The intoxication status caused an intensive formation of free radicals with reduction of the GSH concentration and increase of MDA. Despite the increase in the capacity of plasmatic antioxidant observed with the increase of uric acid and FRAP concentrations, it was not observed any benefit with treatments using oxidant vitamins, isolatedly or together with classic treatment only with TTM.

O objetivo do presente estudo foi avaliar a evolução e as complicações pós-operatórias do tratamento de hérnias perineais bilaterais, por acesso dorsal ao ânus, em 14 cães com idade entre seis e 12 anos. Realizou-se incisão em forma de 'U' invertido, a qual se estendeu desde a porção lateral da tuberosidade isquiática do lado esquerdo, curvando-se em direção à linha mediana, dois centímetros dorsalmente ao ânus e dirigindo-se simetricamente para a extremidade da tuberosidade isquiática contralateral. Após redução da hérnia, os músculos do diafragma pélvico direito e esquerdo foram suturados pela técnica de elevação do músculo obturador interno. As complicações observadas no período pós-operatório foram incontinência fecal, contaminação da ferida, deiscência de pontos da sutura cutânea, incontinência urinária e recidiva em um cão 19 dias após procedimento cirúrgico. O acesso utilizado foi exequível e todos os cães submetidos à técnica operatória proposta não apresentaram recidivas em um período entre cinco e dez meses de avaliação pós-operatório. | Observing the evolution and complications in the treatment of bilateral perineal hernias by dorsal access to the anus was the objective of this study. Fourteen dogs of various breeds were operated with an inverted "U" shaped incision, which extended from the lateral portion of the tuberosity Isquiatica on the left, bending into the direction of the midline, dorsally to anus and heading out symmetrically to the tip of the tuberosity Isquiatica contralateral. After reduction of the hernia, the right and left muscles of the pelvic diaphragm were sutured by the technique of raising the internal obturator muscle. The complications observed in the postoperative period were fecal incontinence, contamination of the wound, dehiscence of suture points of the skin, urinary incontinence and recurrence in a dog with 19 days after surgery. The access used was feasible and all dogs undergoing surgical technical proposal showed no recurrence in a period of five to ten months of postoperative assessment.

Foram realizadas 16 coletas, oito no período chuvoso e oito no período de estio, em quatro fazendas de carcinicultura do Estado do Ceará, nos estuários dos rios Jaguaribe (fazendas A e B) e Acaraú (fazendas C e D), totalizando 32 amostras. O objetivo da pesquisa foi quantificar e identificar Vibrio spp. nas amostras de água e sedimento. Os valores máximos da Contagem Padrão em Placas (CPP) de Vibrio spp. encontrados para as amostras de água, no período chuvoso, foram de 5.10³ UFC/mL est. e, para o sedimento, de 7,5.10³ UFC/g est. No período de estio, a CPP máxima para água foi de 4,35.10² UFC/mL est. e de 3,55.10³ UFC/g est. para as amostras de sedimento. Foram obtidos 36 isolados de Vibrio: Vibrio spp. (17), V. vulnificus B1(3); V. calviensis (2), V. cholerae (2), V. litoralis (2), V. metschnikovii (2), V. agarivorans (1), V. alginolyticus (1), V. campbellii (1), V. coralliilyticus (1), V. diazotrophicus (1), V. logei (1), V. mediterranei (1), V. vulnificus B2 (1). O conhecimento da presença de espécies, nunca anteriormente isoladas em viveiros de fazendas de carcinicultura, tais como o V. coralliilyticus, V. agarivorans, V. litoralis e V. calviensis são importantes para o monitoramento microbiológico contínuo desses ambientes. | Sixteen collections were taken, eight during rainy season and eight on the draught season in four shrimp farms in Ceará State, from Jaguaribe River (farms A and B) and Acaraú River (farms C and D) estuaries, totalizing 32 samples. The goal of the research was to identify and quantify Vibrio spp. in water and in sediment samples. The maximum Standard Plate Count (SPC) values of Vibrio spp. calculated for the rainy season, from water, was 5.10³ CFU/mL est., and for the sediment 7.5.10³ CFU/g est.. For the draught season, maximum counting for water was 4.35.10² CFU/mL est. and for sediment 3.55.10³ CFU/g est.. Thirty six strains of Vibrio were isolated: Vibrio spp. (17), V. vulnificus B1(3), V. calviensis (2), V. cholerae (2), V. litoralis (2), V. metschnikovii (2), V. agarivorans (1), V. alginolyticus (1), V. campbellii (1), V. coralliilyticus (1), V. diazotrophicus (1), V. logei (1), V. mediterranei (1), V. vulnificus B2 (1). The isolation of species never previously isolated of shrimp farms, such as V. coralliilyticus, V. agarivorans, V. litoralis and V. calviensis are important for the continuous microbiological monitoring these environments.

O objetivo desta pesquisa foi determinar a influência do uso da plasmaferese sobre o tempo de recuperação clínica e hematológica de caprinos doadores de sangue total ou plasma. Para tanto, foram utilizados 20 caprinos adultos e clinicamente sadios, distribuídos por dois grupos de dez animais cada, a saber: grupo controle (de animais doadores de sangue total não tratados) e grupo experimental (de animais doadores que foram tratados com a plasmaferese). Os caprinos foram selecionados e monitorados por meio de exames físicos (funções vitais) e complementares (hemograma, proteínas totais, albumina, globulinas, relação A:G, ureia, creatinina e hemoglobina livre no plasma) realizados nos seguintes momentos: imediatamente antes e após a doação de sangue: 12, 24, 72, 120, 240, 360, 480 e 720 horas após os procedimentos. Os resultados foram analisados com comparações dentro e entre os dois grupos nos diferentes momentos do estudo. As observações clínicas efetuadas durante o período de até 30 dias após a doação de 20% do volume sanguíneo total, com ou sem a realização da plasmaferese nos animais dos grupos estudados, não sofreram variações influenciadas por esses procedimentos. Observou-se significativa variação dos componentes do eritrograma, tendo o grupo experimental apresentado as melhores taxas de recuperação em função do tempo. Com base nos resultados obtidos, a aplicação da técnica da plasmaferese em caprinos mostrou-se eficiente como recurso para a otimização do tempo de recuperação dos valores do hemograma de animais doadores de plasma, não determinando hemólise durante o seu procedimento. | The objective of this study was to determine the influence of plasmapheresis on clinical and haematological recovery time of whole blood or plasma donor goats. For this, 20 clinically healthy adult goats were divided into two groups of ten animals each: control group (not-treated whole blood donor animals), and experimental group (donor animals which were treated with plasmapheresis). Goats were selected and evaluated through physical examination (vital functions) and complementary tests (complete blood counts, total proteins, albumin, globulin, albumin:globulin ratio, urea nitrogen, creatinine, and plasma free haemoglobin) were carried out at the following moments: immediately before and after blood donation, 12, 24, 72, 120, 240, 360, 480, and 720 hours after the procedures. Results were analysed comparing animals in and between both groups (at differents moments of the study). The clinical observations made during the period of 30 days after donation of 20% of total blood volume, with or without plasmapheresis in the animals of studied groups, were not influenced by these procedures. The results revealed significant variation of eritrogram components, showing the experimental group to have better recovery rates according to time. Based on the results obtained in the present study, plasmapheresis technique application in goats showed to be efficient as a resource to optimize recovery time of blood cell values of plasma donor animals, and did not cause hemolisis during its procedure.

O exame ultrassonográfico apresenta divergência na interpretação dos seus achados, devido à análise subjetiva e individual. Avaliou-se ultrassonograficamente a quantificação dos níveis de cinza de fígado de 18 gatos hígidos pela técnica de histograma, obtendo-se medidas referentes à ecogenicidade (LMEAN), ecotextura (NMOST/NALL) e desvio padrão da amplitude do eco (SD). As médias e desvio padrão dos valores encontrados foram para LMEAN 5,6 ±1,08, NMOST/NALL 28,31 ±3,84 e SD 11,48 ±1,77. Ressalta-se a importância da técnica do histograma na padronização de valores de normalidade referentes à ecogenicidade e ecotextura do fígado, favorecendo o diagnóstico e o monitoramento de hepatopatias difusas. | The ultrasonographic exam presents divergence in the interpretation of its findings, due to subjective and individual analysis. The livers of 18 healthy cats were evaluated by the ultrasonographic exam, making use of the gray-level histograms method. There were taken measures referring to the echogenicity (LMEAN), echotexture (NMOST/NALL) and echo amplitude standard deviation (SD). The mean values and standard deviations observed in this clinical assay were: LMEAN 5.6 ±1.08, NMOST/NALL 28.31 ±3.84 e SD 11.48 ±1.77. The gray-level histograms method is very important for the standardizing of hepatic echogenicity and echotexture values, helping in the diagnosis and monitoring of diffuse hepatopathies.

As pesquisas em nutrição de frangos de corte estão em busca de ajustes que forneçam às aves os nutrientes necessários para um ótimo desempenho produtivo. Neste contexto, a utilização de microminerais orgânicos vem ganhando força e se mostrando uma alternativa para aumentar a produção. O presente estudo teve como objetivo avaliar os efeitos de três fontes e dois níveis de selênio sobre o desempenho de frangos de corte. Foram utilizados 1440 pintos de um dia, machos, criados até os 42 dias. O delineamento experimental foi inteiramente casualizado com seis dietas experimentais (A: 0,15 mg/kg Se inorgânico; B: 0,15 mg/kg Se orgânico; C: 0,15 mg/kg Se inorg. + orgânico; D: 0,45 mg/kg Se inorgânico; E: 0,45 mg/kg Se orgânico; F: 0,45 mg/kg Se inorg.+ orgânico) e seis repetições com 40 aves cada. Foi utilizado um arranjo fatorial 3 x 2 e os dados obtidos foram analisados pelo Statistical Analysis System. Quanto ao desempenho, considerando-se o período total de criação, houve efeito do nível de Se sobre o ganho de peso (GP) e ganho médio diário (GMD); a conversão alimentar foi a única variável que apresentou interação entre fonte e nível de Se, sendo que as variáveis de peso somente apresentaram efeito do nível de inclusão, mostrando que fonte e nível de Se são fatores que juntos podem exercer influência sobre o desempenho dos animais. | The research in nutrition of broiler chickens is searching for adjustments that supply to the birds the necessary nutrients for better productive performance. In this context the use of organic minerals is gaining force and becoming an alternative to increase the production. The objective of the present study was to evaluate the effect of three sources and two levels of selenium on the performance of broiler chickens. This work analysed 1440 young chickens of one day, males, that were created until the 42nd day. The assignment was completely randomized, with six experimental diets (A: 0,15 mg/kg inorganic; B: 0,15 mg/kg organic; C: 0,15 mg/kg inorg. + organic; D: 0,45 mg/kg inorganic; E: 0,45 mg/kg organic; F: 0,45 mg/kg inorg. + organic) and six repetitions with 40 birds each. It was used a 3 x 2 factorial arrangement and the obtained data were analyzed by PROC GLM of SAS. In regard to the performance, considering the total period (42 days), there was an effect of the level of Se on the weight gain (GP) and average daily gain (GMD); the alimentary conversion was the only variable that presented interaction between source and level of Se and the weight variables only presented the effect of the inclusion level, showing that source and level of Se are factors that together can exercise influence on the performance of the animals.