ABSTRACT: The study aimed to describe the clinicopathological aspects of 37 cases of pythiosis, 34 in horses and three in mules, from properties located in the Amazon biome of Pará, Brazil. The clinical signs observed in the animals were weakness, poor-to-regular nutritional status, pale mucous membranes, itching at the lesion site, and lameness when the limbs were affected. The lesions were located on the lips, nostrils, rib region, thoracic and abdominal walls, scapular, distal limbs, foreskin, perineum and udder. Macroscopically, ulcerative and granulation-tissue-like masses were observed, with fistulous tracts filled with yellowish and foul-smelling serosanguinous discharges. In the biopsy, it was possible to visualize white and firm areas with foci of yellowish necrotic material and hardened masses, called kunkers. The histopathological examination showed a pyogranulomatous inflammatory reaction with the presence of Pythium insidiosum hyphae, which were impregnated with black, confirming the diagnosis of pythiosis in equids in the Amazon biome, being the first report of the disease in mules in the region.

RESUMO: O trabalho objetivou descrever os aspectos clínico-patológicos de 37 casos suspeitos de pitiose, 34 em equinos e três em muares, provenientes de propriedades localizadas no Pará, bioma amazônico brasileiro. Os sinais clínicos observados nos animais eram caracterizados por debilidade, estado nutricional de ruim a regular, mucosas pálidas, prurido no local da lesão, além de claudicação quando os membros foram acometidos. As lesões eram localizadas nos lábios, narinas, região das costelas, parede torácica e abdominal, escapular, distais dos membros, prepúcio, períneo e úbere. Macroscopicamente observavam-se extensas lesões ulceradas com intensa proliferação de tecido de granulação, de bordos irregulares, com tratos fistulosos, de consistência firme, denominados de “kunkers”, preenchidos com material amarelado e friável, possuindo exsudação serossanguinolenta de odor fétido. Ao exame histopatológico observou-se reação inflamatória piogranulomatosa com presença de hifas de Pythium insidiosum, as quais se impregnaram de negro, confirmando o diagnóstico de pitiose em equídeos no Bioma Amazônico, sendo o primeiro relato da doença em muares na região.

ABSTRACT: This study aimed to evaluate phosphorus (P) concentration in serum and bone, the percentage of ash and the specific bone density of buffaloes on Ilha de Marajó before and after mineral supplementation. For this study, 14 crossbred buffaloes of Murrah and Mediterranean descent aged between 18 and 36 months were used. The average values of P before supplementation in serum and bone, the percentage of bone ash and the specific bone density were 5.68mg/dL±1.18, 16.53%±0.53, 59.95%±1.96 and 1.52g/cm3±0.32, respectively, which demonstrated P deficiency in animals raised on Ilha de Marajó. After supplementation with P for a period of seven months, the values were 6.61mg/dL±0.87, 16.90%±0.56 and 60.30%±0.95 and 1.71g/cm3±0.21, respectively. These results showed a significant increase in P concentration in blood serum, specific bone density and percentage of P in ash (P<0.05), but there was no significant increase in the percentage of ash. The average increase in P in the serum and ash did not reach normal levels in all animals; however, 28.6% of the animals had normal values of P in serum and 50% in the ash, and 64.3% had normal specific bone density values. The nonre-establishment, in some of the animals, of the variables of P serum and bone after supplementation for seven months may have occurred as a result of the low intake of the mineral mixture and by the low concentration of P in the Brachiaria brizantha cv. Marandu used for feeding animals during the experiment.

RESUMO: Objetivou-se avaliar as concentrações de fósforo (P) no soro e no osso, o percentual de cinzas e a densidade óssea específica em búfalas da Ilha de Marajó antes e após suplementação mineral seletiva. Foram utilizadas 14 búfalas mestiças de Murrah com Mediterrânea, com idades entre 18 e 36 meses. Os valores médios de P, antes da suplementação, no soro, no osso, o percentual de cinzas e a densidade óssea específica foram de 5,68mg/dL±1,18, 16,53%±0,53, 59,95%±1,96 e 1,52g/cm3±0,32, respectivamente, o que demonstra deficiência de P nos animais criados na Ilha de Marajó. Após a suplementação com P por um período de sete meses os valores foram 6,61mg/dl±0,87, 16,90%±0,56 e 60,30%±0,95 e 1,71g/cm3±0,21 respectivamente. Esses resultados caracterizam um aumento significativo nas concentrações de P no soro sanguíneo, na densidade óssea específica e no percentual de P nas cinzas (P<0,05), porém não houve um aumento significativo no percentual de cinzas. O aumento médio nos valores de P no soro e nas cinzas não alcançou patamares de normalidade em todas as búfalas, entretanto 28,6% delas tinham valores normais de P no soro e 50% nas cinzas, 64,3% tinham valores normais da densidade óssea específica. O não restabelecimento, em parte dos animais, das variáveis ósseas e sanguíneas após suplementação com P durante sete meses pode ter ocorrido em virtude da baixa ingestão da mistura mineral e da baixa concentração de P em Brachiaria brizantha cv. Marandu utilizada para alimentação dos animais durante o experimento.

RESUMO: A dermatite digital bovina (DDB) foi estudada em vacas mestiças leiteiras nos municípios de Rondon do Pará no Pará, Açailândia e Cidelândia no Maranhão, bioma amazônico brasileiro. Para tanto, foram realizadas quatro visitas a 10 propriedades, nos meses de agosto e novembro em 2016 e abril e julho em 2017, e realizada a inspeção dos dígitos de 1.664 vacas em lactação durante a ordenha. Nestas propriedades as vacas eram criadas a pasto durante todo o ano e ordenhadas mecanicamente em oito propriedades. As quais, as salas de ordenha tinham piso concretado, com proteção contra as chuvas e era realizada a limpeza diariamente. No entanto, em três dessas propriedades, as salas de pré ou pós ordenha tinham piso não concretado e em duas, a ordenha era manual em piso de chão batido. Em todas elas não havia medidas profiláticas para afecções podais. A prevalência de DDB foi de 1,3% (22/1.664) e não se obteve diferença estatística entre os períodos chuvoso e não chuvoso (P = 0,72). As lesões observadas foram classificadas de acordo com o sistema M (M0-sem lesão; M1-lesão ulcerada <2cm; M2-lesão ulcerada >2cm; M3-lesão em cicatrização; M4-lesão crônica; M4.1- M4 com área ulcerada). De 22 lesões observadas, 22,7% (5/22) apresentavam-se em estágio M1, 36,4% (8/22) em M2, 22,7% (5/22) em M3, 13,6% (3/22) em M4 e 4,5% (1/22) em M4.1. Pelos hipertrofiados nos bordos das lesões, larvas de moscas do gênero Cochliomyia spp. e alterações no tecido córneo também foram observados. Nas lesões de 11 bovinos, em seis, foi realizado o tratamento tópico com extrato bruto de Copaifera reticulata (óleo de copaíba) e em cinco, com a pasta de ácido salicílico a 660mg/g e ambos os tratamentos foram protegidos com bandagem por sete dias. Após, as lesões foram acompanhadas semanalmente até a cura e obteve-se um índice de 83,4% (5/6) e 75% (3/4) de cura com esses tratamentos, respectivamente, com uma média de 7 semanas. Conclui-se que no bioma amazônico a DDB ocorre em baixa prevalência, não sazonal e com características macroscópicas semelhantes às lesões de vacas leiteiras criadas em sistema free stall. O tratamento com o óleo da copaíba apresentou resultados semelhante ao ácido salicílico e pode ser uma alternativa como medida de controle da DDB no bioma amazônico.

ABSTRACT: Bovine Digital Dermatitis (BDD) was studied in crossbred dairy cows grazing in Rondon do Pará, in the state of Pará, as well as in Açailândia and Cidelândia, in the state of Maranhão, Brazilian Amazon biome. The digits inspection from the dairy cows during milking was performed in ten farms comprising four visits (August and November 2016; April and July 2017). The cows were kept all year in pastures, and were mechanically milked on concrete floors and the animals were protected against the rains in eight farms, maintaining a daily cleaning, however, it could not be found a concrete floor in pre- or post-milking to ensure milking parlor on three farms. Manual milking on no concrete floors was performed in two farms. No preventive measures against hoof lesions were adopted. The BDD prevalence was 1.3% (22/1664), and no statistical difference among rainy or no rainy season was obtained (p = 0.72). The BDD lesions were classified according to “M system” (M0 = no lesion, M1 = active ulceration <2cm, M2 = active ulceration >2cm, M3 = healing stage, M4 = chronic stage, M4.1 = M4 with active ulceration). Regarding the 22 BDD lesions observed, 22.7% (5/22) were M1 stage, 36.4% (8/22) M2, 22.7% (5/22) M3, 13.6% (3/22) M4 and 4.5 (1/22) M4.1. Hypertrophic hairs at the edges of the lesions caused by fly larvae of genus Cochliomyia spp. as well as alterations on the hoof were also observed. Topical treatment was performed in six BDD lesions with a raw extract from trees of the genus Copaifera reticulata (Copaiba oil) and compared with the treatment of salicylic acid paste in five BDD lesions. The lesions were protected with a bandage for seven days and followed weekly until recovery. The complete therapeutic responses were 83.4% (5/6) and 75% (3/4), respectively, with an average time of seven weeks. The BDD in the Amazon biome occurs in low prevalence, not seasonal, and presents macro and microscopic features similar to BDD lesions from dairy cattle kept in free-stall housing. The treatment with copaiba oil showed similar results to the treatment of salicylic acid paste and can be used in control measures to BDD in the Amazon biome.

RESUMO Estruturas oculares de cães braquicéfalos foram estudadas por elastografia acoustic radiation force impulse (ARFI) para estabelecer valores quantitativos e qualitativos de referência relacionados com a forma e a rigidez de algumas estruturas oculares. Foram avaliados 84 olhos de 42 cães braquicéfalos neste estudo. Os animais foram submetidos aos exames físico, laboratorial e aos exames oftalmológicos. Após a confirmação da sanidade dos pacientes, os cães foram submetidos à ultrassonografia e à elastografia ARFI, enfatizando-se o nervo óptico, o corpo ciliar e a lente. Para elastografia qualitativa, foram observadas características homogêneas no corpo ciliar e no nervo óptico. Os humores aquoso e vítreo foram visibilizados como imagens tipo mosaico. A elastografia quantitativa demonstrou velocidades de cisalhamento para nervo óptico de 1,01±0,27m/s, para corpo ciliar temporal de 0,91±0,24m/s e para corpo ciliar nasal de 0,91±0,3m/s. Valores dessas velocidades para lentes apresentaram-se fora do intervalo: X.XXm/s. A elastografia provou ser um procedimento não invasivo e viável para cães. Estabelecidos os parâmetros do grau de rigidez das estruturas oculares, futuramente tais dados servirão como padrão para avaliação de animais com os distúrbios oculares. Os resultados podem ser extrapolados para a pesquisa primária sobre a aplicabilidade da ARFI na avaliação do bulbo ocular em seres humanos.

ABSTRACT Eyes from brachycephalic dogs were studied by acoustic radiation force impulse (ARFI) elastography to establish quantitative and qualitative reference values related to the shape and rigidity of some ocular structures. Eighty-four eyes from 42 adult brachycephalic dogs were included in this study. Animals were subjected to physical, laboratory, and ophthalmic examinations. Following confirmation of good health, the dogs were subjected to ultrasonography and ARFI elastography, emphasizing the optic nerve, ciliary body, and lens. On qualitative elastography, homogeneous characteristics were observed on the ciliary body and optic nerve. Aqueous and vitreous humors were visualized as mosaic-like images. Quantitative elastography showed the mean optic nerve shear wave velocity (SWV) was 1.01±0.27m/s, temporal ciliary body was 0.91±0.24m/s, and nasal ciliary body was 0.91±0.3m/s. The SWV of the lens values were out of range, this value was not detectable by this software. Elastography has proved to be a non-invasive procedure and feasible in dogs. The establishmentof the parameters of degree of rigidity of ocular structures will serve as a baseline for animals with eye disorders. Results may be extrapolated to primary research on the applicability of ARFI in the evaluation of ocular bulb in humans.

RESUMO: Foram avaliados os fatores epidemiológicos e as afecções podais de bovinos de corte no estado do Pará em 12 propriedades. Em todas foi realizado o estudo epidemiológico e em nove o estudo epidemiológico e o exame clínico dos animais. Nos centros de manejo foram observados fatores favoráveis a traumatismos nos dígitos dos bovinos, como piso das seringas calçado com pedras, troncos com exposição de ferragens e rampas dos embarcadores com pisos de pedras pontiagudas, além da falta de manutenção e erros estruturais. Em todas as fazendas observou-se um manejo inadequado dos animais. Em 91,7% das fazendas (11/12) havia piquetes com presença de troncos de árvores e 66,7% (8/12) apresentavam áreas de relevo inclinado com presença de pedras. Em 16,7% (2/12) das fazendas havia áreas de brejo com áreas alagadas. A sodomia foi relatada em todas as propriedades. Nenhuma fazenda adotava medidas profiláticas relacionadas às afecções podais. Foram examinados 498 bovinos entre fêmeas e machos. Os membros pélvicos foram mais acometidos, tanto nas fêmeas quanto nos machos. Foram diagnosticadas 629 lesões nas fêmeas, sendo as mais frequentes pododermatite séptica, deformações ungulares, pododermatite da sobreunha e dermatite digital. Nos machos diagnosticou-se 285 lesões e as mais frequentes foram pododermatite séptica, deformações ungulares e abrasão de talão. Conclui-se que erros de manejo associados às condições inadequadas das instalações foram fatores que contribuíram para a ocorrência de enfermidades podais em bovinos de corte nas fazendas estudadas; as características ambientais favoreceram o desenvolvimento das lesões; as associações de lesões foram mais prevalentes do que as simples, sendo diagnosticadas em animais em todas as fazendas estudadas; o diagnóstico precoce a campo não era realizado, o que contribuiu para aumentar a gravidade e a diversificação das afecções podais diagnosticadas.

ABSTRACT: The epidemiological study was conducted in 12 farms with the history of foot disease occurrence. Clinical examination of the affected cattle for the diagnosis of foot disease was performed only on nine of these farms. Multiple risk factors were found for digit injuries, as syringes floors paved with rocks, working chutes with iron fittings exposure, loading ramps covered with sharp stones and the lack of maintenance and structural errors. On each farm there was an inappropriate handling of cattle. On 91.7% of them (11/12) existed pickets with trunks of trees and 66.7% (8/12) had areas of sloping relief with stones. Moreover, 16.7% (2/12) of the farms were in marsh areas and wetlands. Sodomy was reported for all farms. None of them adopted preventive measures related to foot problems. A total of 498 cattle, males and females, were examined. The hind limbs were the most affected in both, females and in males. In females 629 lesions were diagnosed. The septic pododermatitis was the most common, followed by claw deformities, pododermatitis in the paradigits and digital dermatitis. In males 285 injuries were diagnosed; the most common were septic pododermatitis, claw deformities and heel erosion. It was concluded that management errors associated with inadequate facilities were factors that contributed to the occurrence of foot diseases in beef cattle of the farms studied. The environmental characteristics favored the development of lesions; combined injuries were more prevalent than the simple ones diagnosed in cattle of all farms. Early diagnosis was not carried out in the field, what helped to increase the severity and the diversification of the foot problems.

ResumoRealizou-se um estudo das deficiências minerais em vacas em lactação de rebanhos leiteiros pertencentes a 13 propriedades da bacia leiteira do município de Rondon do Pará, estado do Pará. Foram determinados os níveis de fósforo (P) no osso, e os níveis de cobre (Cu), cobalto (Co), selênio (Se) e zinco (Zn) no fígado de 47 vacas leiteiras no 2º terço da lactação. Estas amostras foram coletadas por meio de biópsias realizadas no terço superior da 12a costela do lado direito e no bordo caudal do lobo caudado do fígado, respectivamente. Os rebanhos eram formados por animais mestiços (Holandes x Zebu), mantidos em sistema de produção extensivo em pastos de Brachiaria brizantha cv Marandu e recebiam suplementação mineral. A mistura mineral em 12 propriedades era do tipo comercial, dita "completa", acrescida de quantidades de NaCl acima do recomendado pelos fabricantes em dez propriedades. Em sete propriedades as misturas minerais eram fornecidas em cochos sem cobertura e em oito, o fornecimento da mistura mineral não era realizado diariamente. Em 11 propriedades, havia históricos clínicos condizentes com deficiências minerais nos rebanhos. Nessas fazendas a retenção de placenta e a osteofagia foram as alterações mais relatadas. Após as análises minerais observou-se deficiência de P em cinco propriedades, de Co em três propriedades, de Se em nove propriedades e de Zn em dez propriedades. Conclui-se que ocorre a deficiência de P, Co, Se e Zn; a suplementação mineral realizada na maioria das propriedades não atendeu as exigências diárias de P, Se e Co, baseadas no consumo estimado de 30 g de NaCl/animal/dia; os cochos pouco adequados ou inadequados para a suplementação, assim como o fornecimento inconstante das misturas minerais possivelmente contribuíram para a deficiência de um ou mais minerais.

AbstractA study of mineral deficiencies was performed in lactating cows from dairy herds belonging to 13 farms of Rondon do Pará, state of Pará, Brazil. We determined the levels of phosphorus (P) in the bone and levels of copper (Cu), cobalt (Co), selenium (Se) and zinc (Zn) in the liver of 47 dairy cows in lactation 2. The samples were collected by means of biopsies taken on the top third of the 12th rib of the right side and the caudal edge of the caudate liver lobe, respectively. The herds were composed of crossbred (Holstein x Zebu) cattle, kept in extensive production system on Brachiaria brizantha cv. Marandu pasture and received mineral supplementation. The mineral mixture on 12 farms were commercial type, called "full" and the owners added a quantity of salt above the manufacturer's recommendations on ten farms. The mineral mixtures were provided in troughs without covering on seven farms and on other eight, providing of the mineral mix was not performed daily. On 11 farms there were clinical histories consistent with mineral deficiencies in livestock, and placenta retention and osteophagia were the most frequently reported ones. The results of chemical analysis showed deficiency of P on five farms, of Co on three farms, of Se on nine farms and of Zn on ten farms. It is concluded that the mineral supplementation performed on the farms do not supply the daily demands of P, Se and Co, based on the estimated daily consumption of 30g of NaCl per animal; the little adequate or inadequate troughs for proper supplementation, as well as inconstant supply of mineral mixtures contribute to the deficiency of one or more minerals.

A padronização de técnicas e o teste da associação do Plasma Rico em Plaquetas (PRP) e das Células-Tronco Mononucleares (CTMs) na consolidação de falhas ósseas corticais, por meio de avaliação clínica, biomecânica, radiológica e histológica, é avaliada em um estudo piloto. Foram utilizados seis cães adultos, fêmeas, sem raça definida, pesando entre 5 e 10kg, separados por sorteio aleatório em seis tratamentos. Foi confeccionada uma falha elíptica de 1,0x0,4cm na cortical medial diafisária da tíbia direita de cada animal, sendo preenchida de acordo com o tratamento proposto. No cão I, a falha foi preenchida com solução fisiológica (SF); no II, com o PRP; no III, com a fração total das células mononucleares (FTCM); no IV, com a fração vascular estromal (FVE); no V, com o PRP associado à FTCM; no VI, com a associação PRP e FVE. Foram realizadas avaliações: clínicas, diariamente; dos graus de claudicação, semanalmente; radiológica e perimetria da coxa, antes, no pós-operatório imediato, aos 7, 14, 21 e 30 dias; biomecânica, antes do procedimento, aos 10, 20 e 30 dias; e biópsias, aos 15 e 30 dias. A FTCM obteve uma contagem e viabilidade média de 2,0x108cél. e 90%, respectivamente, enquanto a FVE obteve 3x106cél. e 50%. O PRP concentrou, em média, sete vezes o número inicial de plaquetas do sangue total, de 250.000 µl-1 plaquetas no sangue total para 1.750.000 µl-1 plaquetas no PRP. Obteve-se padronização adequada de técnicas, possibilitando o teste da associação entre as células-tronco mononucleares (CTMs) e o plasma rico em plaquetas (PRP), assim como seu uso isolado, no reparo de falhas ósseas corticais, indicando a possibilidade de a associação FTCM e PRP ser o melhor tratamento.

The standardization of techniques and tests of the association of Platelet Rich Plasma (PRP) and Mononuclear Stem Cells (MSCs) in the consolidation of cortical bone defects by clinical, biomechanical, radiological, and histological analysis is evaluated in a pilot study. Six adult female dogs of mixed breed, weighing between 5 and 10kg, separated by random draw in six treatments were used. An elliptical failure of 1.0 x0.4cm was done in the medial diaphyseal cortical of the right tibia of each animal, that was filled according with the proposed treatment. In dog I, the failure was filled with saline (S), in dog II with PRP, in dog III with total mononuclear cell fraction (TMCF), in dog IV with stromal vascular fraction (SVF), in dog V with association of PRP and TMCF, and in dog VI with an association of PRP and SVF. Daily clinical evaluation, weekly degrees of lameness, radiological and girth before, immediate postoperative, 7, 14, 21 and 30 days, biomechanics before the procedure, at 10, 20 and 30 days, and biopsies at 15 and 30 days were performed. The TMCF got a count and viability of 2,0x108cells and 90% respectively, while for SVF it was 3x106cells and 50%, respectively. The PRP concentrated on average seven times the original number of platelets from whole blood, platelets from whole blood 250.000 μl-1to 1.750.000 μl-1platelets in PRP. This afforded adequate standardization of techniques, enabling the test of association between mononuclear stem cells (MSCs) and platelet-rich plasma(PRP), as well as their separate use to repair cortical bone defects, indicating the possibility of the association between FTCM and PRP to be the best treatment.

We encapsulated cisplatin into stealth pH-sensitive liposomes and studied their stability, cytotoxicity and accumulation in a human small-cell lung carcinoma cell line (GLC4) and its resistant subline (GLC4/CDDP). Since reduced cellular drug accumulation has been shown to be the main mechanism responsible for resistance in the GLC4/CDDP subline, we evaluated the ability of this new delivery system to improve cellular uptake. The liposomes were composed of dioleoylphosphatidylethanolamine (DOPE), cholesteryl hemisuccinate (CHEMS), and distearoylphosphatidylethanolamine-polyethyleneglycol 2000 (DSPE-PEG2000) and were characterized by determining the encapsulation percentage as a function of lipid concentration. Among the different formulations, DOPE/CHEMS/DSPE-PEG liposomes (lipid concentration equal to 40 mM) encapsulated cisplatin more efficiently than other concentrations of liposomes (about 20.0%, mean diameter of 174 nm). These liposomes presented good stability in mouse plasma which was obtained using a 0.24-M EDTA solution (70% cisplatin was retained inside the liposomes after 30 min of incubation). Concerning cytotoxic effects, they are more effective (1.34-fold) than free cisplatin for growth inhibition of the human lung cancer cell line A549. The study of cytotoxicity to GLC4 and GLC4/CDDP cell lines showed similar IC50 values (approximately 1.4 µM), i.e., cisplatin-resistant cells were sensitive to this cisplatin formulation. Platinum accumulation in both sensitive and resistant cell lines followed the same pattern, i.e., approximately the same intracellular platinum concentration (4.0 x 10-17 mol/cell) yielded the same cytotoxic effect. These results indicate that long-circulating pH-sensitive liposomes, also termed as stealth pH-sensitive liposomes, may present a promising delivery system for cisplatin-based cancer treatment. This liposome system proved to be able to circumvent the cisplatin resistance, whereas it was not observed when using non-long-circulating liposomes composed of phosphatidylcholine, phosphatidylserine, and cholesterol.

Toxoplasma gondii is an obligatory intracellular parasite whose life cycle may include man as an intermediate host. More than 500 million people are infected with this parasite worldwide. It has been previously reported that T. gondii contains a superantigen activity. The purpose of the present study was to determine if the putative superantigen activity of T. gondii would manifest towards human T cells. Peripheral blood mononuclear cells (PBMC) from individuals with no previous contact with the parasite were evaluated for proliferation as well as specific Vß expansion after exposure to Toxoplasma antigens. Likewise, PBMC from individuals with the congenital infection were evaluated for putative Vß family deletions in their T cell repertoire. We also evaluated, over a period of one year, the PBMC proliferation pattern in response to Toxoplasma antigens in patients with recently acquired infection. Some degree of proliferation in response to T. gondii was observed in the PBMC from individuals never exposed to the parasite, accompanied by specific Vß expansion, suggesting a superantigen effect. However, we found no specific deletion of Vß (or Valpha) families in the blood of congenitally infected individuals. Furthermore, PBMC from recently infected individuals followed up over a period of one year did not present a reduction of the Vß families that were originally expanded in response to the parasite antigens. Taken together, our data suggest that T. gondii does not have a strong superantigen activity on human T cells.