Cytotoxicity of isolated compounds from Picrasma crenata (Vell) Engl. in animal tumor cell (HTC) / Citotoxicidade de compostos isolados de Picrasma crenata (Vell) Engl. em células tumorais animais (HTC)

The study aim was to evaluate the cytotoxic activity, using the MTT test [3-(4,5-Dimethilthiazol-2-yl)-2,5-diphenil tetrazolium bromide], from the crude extract of Picrasma crenata (Pau Tenente) and its isolated compounds, quassin and parain, in culture of rat liver tumor cells (HTC). The test was carried out exposing the cells for 24, 48 and 72 hours to concentrations of 5, 10, 50, 100, 200, 300, 400, 500 and 1000 µg of crude extract of Pau Tenente/mL of culture medium and 1, 5, 10, 15, 20, 40, 60, 80 and 100 µg of quassin or parain compounds/mL of culture medium. The absorbances averages results obtained showed that the crude extract did not present cytotoxicity for the HTC cells in all the concentrations and evaluated times. For quassin, the concentrations of 80 and 100 µg/mL were cytotoxic, after 72 hours of treatment. For parain, the concentrations of 1, 5, 20, 40, 60, 80 and 100 µg/mL, in 72 hours, were cytotoxic, revealing a new activity for this compound. Thus, the results demonstrate a first indication of the cytotoxic activity of compounds quassin and parain, adding an important social and economic value to them, and may have application in future research and in pharmaceutical industry.

O objetivo do estudo foi avaliar a atividade citotóxica, por meio do teste MTT [brometo de 3-(4,5-dimetiltiazol-2-il)-2,5-difenil tetrazólio], do extrato bruto de Picrasma crenata (Pau Tenente) e seus compostos isolados, quassina e paraína, em cultura de células tumorais de fígado de rato (HTC). O teste foi realizado expondo as células por 24, 48 e 72 horas às concentrações de 5, 10, 50, 100, 200, 300, 400, 500 e 1000 µg de extrato bruto de Pau Tenente/mL de meio de cultura e 1, 5, 10, 15, 20, 40, 60, 80 e 100 µg de compostos de quassina ou paraína/mL de meio de cultura. Os resultados das médias de absorbâncias obtidos mostraram que o extrato bruto não apresentou citotoxicidade para as células HTC em todas as concentrações e tempos avaliados. Para quassina, as concentrações de 80 e 100 µg/mL foram citotóxicas, após 72 horas de tratamento. Para a paraína, as concentrações de 1, 5, 20, 40, 60, 80 e 100 µg/mL, em 72 horas, foram citotóxicas, revelando uma nova atividade para este composto. Assim, os resultados demonstram uma primeira indicação da atividade citotóxica dos compostos quassina e paraína, agregando-lhes importante valor social e econômico, podendo ter aplicação em pesquisas futuras e na indústria farmacêutica.

The effect of adding L-ergothioneine in culture medium on blastocyst development rates

Background: Oocytes and embryos produce energy through mitochondrial oxidative phosphorylation by using oxygen. The membrane structure of the embryo is mostly composed of unsaturated fatty acids, for this reason DNA fragmentation, apoptosis, and abnormal gene expression are shaped as a result of the lipid peroxidation during culture. Oxidative stress (OS) is one of the most important problems affecting the in vitro embryo development. Antioxidant supplementation to the culture medium has been an alternative way to reduce cell damage caused by oxidative stress in in vitro embryo production systems. In this study, it was aimed to determine the effect of L-ergothioneine on blastocyst development when added to the culture medium. Materials, Methods & Results: The material of the study consisted of oocytes aspirated from the ovaries of Holstein cows which were collected from the local slaughterhouse. The ovaries were delivered to the laboratory within 2-3 h in a thermos which provided a constant temperature of 25-30o C with physiological saline (0.9%) containing antibiotics. All follicles in the 3-8 mm range on the ovaries were aspirated using 20 G needle. The collected follicle fluid was filtered through filters with a pore diameter of 70 micrometers. Cells remaining in the filter were washed with OPU medium and transferred to the petri dishes. Fluids were examined under a stereomicroscope. The cumulus-oocyte complexes were classified, and A and B quality oocytes were included to the study (A, B, C, and D quality COC). Oocytes aspirated from the ovaries and collected later on were incubated in IVM medium for 22 h. After maturation, it was taken into IVF medium, semen was added and incubated for 20-22 h. Possible zygotes to be taken to the culture stage were transferred to culture (IVC) drops with (L-ergothioneine 100 µL/mL (n:121) added and without antioxidant (control (n:124)), and kept in the incubator for 6-7 days. Evaluated on the 7th day differences in in vitro embryo production stages were evaluated with the Chi-square test. The study was run in 5 replicates each time, with at least 20 possible zygotes for per group being cultured. It was determined that 262 (87.33%) of a total of 300 oocytes undergoing in vitro maturation were matured. It was determined that 245 of the mature oocytes were fertilized (93.51%). The cleavage rates of the groups were determined as 87.60% and 86.29%, respectively. Eighty-two (33.47%) blastocysts were obtained from 245 zygotes taken into the culture stage, and the blastocyst rates in the groups were found to be 40.50% and 26.61%, respectively. After the study, it was determined that the statistical difference between L-ergothioneine and control in cleavage rates was insignificant (P > 0.05) and blastocyst rates was significant (P < 0.05) Discussion: Oxygen content above normal ratios can increase the formation of reactive oxygen species (ROS), particularly hydrogen peroxide (H2 O2 ), hydroxyl radical (HO·), and peroxyl radicals (ROO·). The increased rate of ROS negatively affects the success of IVP in mammalian embryos. It was observed that L-ergothioneine, which has high antioxidant activity, improved blastocyst development rates, and higher blastocyst rates could be achieved compared to the control group. By investigating the use of L-ergothioneine in different doses, it was thought that the dose with the highest antioxidant activity could be added to the culture medium in in vitro embryo production and more blastocysts could be produced.

Turmeric ethanol extract (Curcuma longa L. ) reduces apoptosis and promotes canine osteosarcoma cell proliferation / Extrato etanólico de açafrão (Curcuma longa L. ) reduz apoptose e promove proliferação de células de osteossarcoma canino

Curcuma longa L., also known as turmeric, has been widely studied for its various therapeutic properties, including antineoplastic action. The ethanolic extract of the plant contains several phenolic compounds, especially curcumin. Osteosarcoma is a predominant bone tumor in dogs and humans, characterized by high metastatic potential and an unfavorable prognosis. The aim of this study was to investigate the effects of turmeric ethanol extract on canine osteosarcoma cells from established culture. The cells were cultured and treated with different curcumin concentrations (0, 10 µM, 20 µM, 50 µM, 100 µM, and 1000 µM) and exposure times (24h, 48h, and 72h). We first performed tetrazolium reduction technique (MTT) assay and calculated IC50. An immunocytochemistry assay was performed after extract treatment to verify the expression of mutated p53 and therefore study the proliferative potential of malignant cells; Bcl-2 and Ki-67 were used to assess apoptosis and the degree of malignancy, respectively. The extract enhanced the proliferation of canine osteosarcoma cells, reaching 3,819.74% at 50 µM of curcumin. The extract also significantly altered the expression of mutated p53 and Ki-67 proteins but not that of Bcl-2, suggesting that it did not induce this antiapoptotic pathway. Overall, these results are prerequisite to better understanding how natural compounds such as turmeric ethanolic extract affect cell proliferation and could be used to treat various diseases.

A Curcuma longa L., planta conhecida popularmente como açafrão, tem sido amplamente estudada por suas diversas propriedades terapêuticas, incluindo a ação antineoplásica. O extrato etanólico da planta contém diversos compostos fenólicos, com destaque para a curcumina. O osteossarcoma é um tumor ósseo predominante em cães e humanos, caracterizado por apresentar alto potencial metastático e prognóstico desfavorável. Procurou-se investigar os efeitos de diferentes concentrações de curcumina do extrato etanólico de açafrão sobre células de osteossarcoma canino de cultura estabelecida. As células foram cultivadas e submetidas ao tratamento com extrato com diferentes concentrações de curcumina (0, 10 µM, 20 µM, 50 µM, 100 µM e 1000 µM) e tempos de exposição (24h, 48h e 72h) pelo EEA. Inicialmente, foram realizados: técnica de redução do tetrazólio (MTT) e cálculo da IC50. Posteriormente, após o tratamento com o extrato, realizou-se o ensaio de imunocitoquímica para verificar a expressão de p53 mutada e estudar o potencial proliferativo das células malignas; Bcl-2, com intuito de averiguar o estímulo de via antiapoptótica; e o marcador Ki-67, que sinaliza aumento no grau de malignidade. O extrato promoveu proliferação de células de osteossarcoma canino, com incremento de até 3819,74% na concentração de 50µM de curcumina. O composto também alterou a expressão das proteínas p53 mutante e Ki-67 significativamente, mas não alterou a expressão de Bcl-2, mostrando que não induziu a via antiapoptótica mediada por esta. Estes resultados demonstram que o extrato etanólico do açafrão apresenta potencial proliferativo sobre células de osteossarcoma canino, sugerindo a necessidade de conscientização e conhecimento dos reais efeitos de determinados compostos naturais, considerados seguros ao serem utilizados como tratamento de diversas enfermidades.

The role of Rho-associated kinase inhibitor, Y-27632 on primary culture of ovine spermatogonial stem cells

Abstract The access to sufficient numbers of spermatogonial stem cells (SSCs) is a prerequisite for the study of their regulation and further biomanipulation. Rho kinase (ROCK) belongs to a family of serine/threonine kinases and involves in a wide range of fundamental cellular functions. The aim of the present study was to study the effect of ROCK inhibitor, Y-27632 (0.1-40 µM), during the primary culture of ovine SSCs. SSCs were collected from 3-5-month-old's lamb testes. The viability of SSCs, the apoptosis assay of SSCs, the intracellular reactive oxygen species (ROS) analysis, and the SSCs markers and apoptosis-related gene expressions were detected by MTT reduction assay, Annexin V-FITC/ Propidium Iodide (PI) dual staining, flow cytometry and real-time-PCR studies, respectively. Morphological analyses indicated that the 5-10 µM Y-27632 had an optimal effect on the number of presumptive SSCs colonies and the area covered by them after a 10 days culture. The cell viability, apoptosis and necrosis of SSCs after 10 days' culture were not affected in comparison with the control group, and the 20 µM of Y-27632 resulted in significantly decreased cell viability (P<0.05) and an increased necrosis of cells. On day 10 after culture, the expression of P53 was decreased with an increase from 0 to 10 µM in the Y-27632 dose. In the 20 µM Y-27632 group, the expressions of P53 and Bax were higher and the Bcl-2 was lower than other groups and these values were significantly different from 5 and 10 µM Y-27632 groups (P<0.05). The level of intracellular ROS was decreased with an increase in the Y-27632 dose from 5 to 20 µM in comparison with the control group. In conclusion, the present study demonstrated that Y-27632 at a concentration of 5-10 µM provided optimal culture conditions for the primary culture of ovine SSCs.

The role of Rho-associated kinase inhibitor, Y-27632 on primary culture of ovine spermatogonial stem cells

The access to sufficient numbers of spermatogonial stem cells (SSCs) is a prerequisite for the study of their regulation and further biomanipulation. Rho kinase (ROCK) belongs to a family of serine/threonine kinases and involves in a wide range of fundamental cellular functions. The aim of the present study was to study the effect of ROCK inhibitor, Y-27632 (0.1-40 µM), during the primary culture of ovine SSCs. SSCs were collected from 3-5-month-olds lamb testes. The viability of SSCs, the apoptosis assay of SSCs, the intracellular reactive oxygen species (ROS) analysis, and the SSCs markers and apoptosis-related gene expressions were detected by MTT reduction assay, Annexin V–FITC/ Propidium Iodide (PI) dual staining, flow cytometry and real-time-PCR studies, respectively. Morphological analyses indicated that the 5-10 µM Y-27632 had an optimal effect on the number of presumptive SSCs colonies and the area covered by them after a 10 days culture. The cell viability, apoptosis and necrosis of SSCs after 10 days culture were not affected in comparison with the control group, and the 20 µM of Y-27632 resulted in significantly decreased cell viability (P<0.05) and an increased necrosis of cells. On day 10 after culture, the expression of P53 was decreased with an increase from 0 to 10 µM in the Y-27632 dose. In the 20 µM Y-27632 group, the expressions of P53 and Bax were higher and the Bcl-2 was lower than other groups and these values were significantly different from 5 and 10 µM Y-27632 groups (P<0.05). The level of intracellular ROS was decreased with an increase in the Y-27632 dose from 5 to 20 µM in comparison with the control group. In conclusion, the present study demonstrated that Y-27632 at a concentration of 5-10 µM provided optimal culture conditions for the primary culture of ovine SSCs.(AU)

Performance of potato clones for processing in subtropical and high-altitude tropical regions of Brazil / Seleção de clones de batata para processamento em regiões do Brasil de clima subtropical e tropical de altitude

This work evaluated the performance of potato clones for some agronomic and processing traits, in subtropical and high-altitude tropical regions, under conventional and organic production systems. Experiments were conducted in spring of 2017 and 2018 in Canoinhas-SC (subtropical), under the conventional system, and in winter of 2018 and 2019 in Brasília-DF (tropical), Brazil, under conventional and organic systems. Eleven advanced clones and cultivars Agata (fresh), Atlantic (chipping) and Asterix (French fry) were evaluated. RCBD with tree replications was used, and tuber yield and processing traits, plant vigor and cycle were evaluated. It was not possible to select high yielding and good processing quality clones with short cycle and less vigorous plants. It because higher yield and larger tuber size were attained by longer cycle genotypes, which had higher specific gravity, but darker frying color. The most outstanding clones differed with region and production system. For chipping, in the subtropical region, no clone had adequate quality, but in tropical F177-11-20 presented similar processing traits and higher or similar yield to 'Atlantic' in both systems, and also ORG7785 in organic. Comparing with ‘Asterix, for French fry, in the subtropical region, superior or similar to the standard this cultiva rwere F129-10-06 and F163-11-01; while in the tropical region and conventional system, were the clones ORG7785, F129-10-06, F163-11-01, F177-11-20, ORG6464, ORG 6377, F05-11-03, Odone 80-02 and F174-11-01; and in organic system, the best clones were F54-11-03, ORG7785, F129-10-06, F163-11-01, F177-11-20 and ORG6464.(AU)

Neste trabalho foi avaliado o desempenho de clones de batata para caracteres agronômicos e de qualidade de processamento, em regiões subtropical e tropical de altitude, sob sistema convencional e orgânico de produção. Os experimentos foram conduzidos na primavera de 2017 e 2018 em Canoinhas-SC (subtropical), sobre sistema convencional, e nos invernos de 2018 e 2019 em Brasília-DF (tropical), Brasil, sob sistemas convencional e orgânico. Foram avaliados onze clones avançados e as cultivares Agata (mercado fresco), Atlantic (chips) e Asterix (palitos fritos). Foi utilizado delineamento em blocos casualisados com três repetições. Foram avaliados caracteres relacionados ao rendimento de tubérculos, qualidade de fritura, vigor de planta e ciclo vegetativo. Foi verificado que não foi possível selecionar clones com elevado rendimento de tubérculos e boa qualidade de fritura e com ciclo vegetativo precoce e com plantas menos vigorosas. Isso porque maior rendimento e maiores tamanhos de tubérculos foram associados com ciclo vegetativo mais longo, maior peso específico de tubérculos, porém cor mais escura de fritura. Os melhores clones diferiram com a região e sistemas de produção. Para chips, n aregião subtropical, nenhum clone apresentou bom desempenho, porém no ecossistema tropical, F177-11-20 apresentou similares qualidades industriais e maior ou superior produtividade em comparação com 'Atlantic' em ambos os sistemas de cultivo, também o clone ORG7785 no sistema orgânico. Para o mercado de fritura de palitos, portanto comparando com ‘Asterix, na região subtropical, os clones F129-10-06 eF163-11-01foram superiores ou equivalentes a esta cultivar; enquanto que na região tropical em sistema convencional destacam-se os clonesORG7785, F129-10-06, F163-11-01, F177-11-20, ORG6464, ORG 6377, F05-11-03, Odone 80-02 eF174-11-01; e nos sistema orgânico os clonesF54-11-03, ORG7785, F129-10-06, F163-11-01, F177-11-20 eORG6464.(AU)

Performance of potato clones for processing in subtropical and high-altitude tropical regions of Brazil / Seleção de clones de batata para processamento em regiões do Brasil de clima subtropical e tropical de altitude

This work evaluated the performance of potato clones for some agronomic and processing traits, in subtropical and high-altitude tropical regions, under conventional and organic production systems. Experiments were conducted in spring of 2017 and 2018 in Canoinhas-SC (subtropical), under the conventional system, and in winter of 2018 and 2019 in Brasília-DF (tropical), Brazil, under conventional and organic systems. Eleven advanced clones and cultivars Agata (fresh), Atlantic (chipping) and Asterix (French fry) were evaluated. RCBD with tree replications was used, and tuber yield and processing traits, plant vigor and cycle were evaluated. It was not possible to select high yielding and good processing quality clones with short cycle and less vigorous plants. It because higher yield and larger tuber size were attained by longer cycle genotypes, which had higher specific gravity, but darker frying color. The most outstanding clones differed with region and production system. For chipping, in the subtropical region, no clone had adequate quality, but in tropical F177-11-20 presented similar processing traits and higher or similar yield to 'Atlantic' in both systems, and also ORG7785 in organic. Comparing with ‘Asterix’, for French fry, in the subtropical region, superior or similar to the standard this cultiva rwere F129-10-06 and F163-11-01; while in the tropical region and conventional system, were the clones ORG7785, F129-10-06, F163-11-01, F177-11-20, ORG6464, ORG 6377, F05-11-03, Odone 80-02 and F174-11-01; and in organic system, the best clones were F54-11-03, ORG7785, F129-10-06, F163-11-01, F177-11-20 and ORG6464.

Neste trabalho foi avaliado o desempenho de clones de batata para caracteres agronômicos e de qualidade de processamento, em regiões subtropical e tropical de altitude, sob sistema convencional e orgânico de produção. Os experimentos foram conduzidos na primavera de 2017 e 2018 em Canoinhas-SC (subtropical), sobre sistema convencional, e nos invernos de 2018 e 2019 em Brasília-DF (tropical), Brasil, sob sistemas convencional e orgânico. Foram avaliados onze clones avançados e as cultivares Agata (mercado fresco), Atlantic (chips) e Asterix (palitos fritos). Foi utilizado delineamento em blocos casualisados com três repetições. Foram avaliados caracteres relacionados ao rendimento de tubérculos, qualidade de fritura, vigor de planta e ciclo vegetativo. Foi verificado que não foi possível selecionar clones com elevado rendimento de tubérculos e boa qualidade de fritura e com ciclo vegetativo precoce e com plantas menos vigorosas. Isso porque maior rendimento e maiores tamanhos de tubérculos foram associados com ciclo vegetativo mais longo, maior peso específico de tubérculos, porém cor mais escura de fritura. Os melhores clones diferiram com a região e sistemas de produção. Para chips, n aregião subtropical, nenhum clone apresentou bom desempenho, porém no ecossistema tropical, F177-11-20 apresentou similares qualidades industriais e maior ou superior produtividade em comparação com 'Atlantic' em ambos os sistemas de cultivo, também o clone ORG7785 no sistema orgânico. Para o mercado de fritura de palitos, portanto comparando com ‘Asterix’, na região subtropical, os clones F129-10-06 eF163-11-01foram superiores ou equivalentes a esta cultivar; enquanto que na região tropical em sistema convencional destacam-se os clonesORG7785, F129-10-06, F163-11-01, F177-11-20, ORG6464, ORG 6377, F05-11-03, Odone 80-02 eF174-11-01; e nos sistema orgânico os clonesF54-11-03, ORG7785, F129-10-06, F163-11-01, F177-11-20 eORG6464.

Different tray cell volumes for lettuce grown in conventional and hydroponic system / Diferentes volumes de célula da bandeja para alface cultivada em sistema convencional e hidropônico

Nowadays, an important economic movement is involved in the improvement of polyethylene trays for the production of seedlings, being developed with different volumes, numbers and cell formats in an each culture-specific manner. The objective of the present study was to evaluate the performance of curly lettuce originated from seedlings from trays with different volumes, through the comparison between conventional and hydroponic-NFT cultivation. Five volumes of trays (10, 20, 30, 40 and 50 cm³ of cell-1) were tested in conventional and hydroponic cultivation, from February to March 2016. For the purpose of comparison, the plants were evaluated through the characteristics of height, circumference, stem diameter, number of leaves, fresh shoot mass and productivity. The use of volumes greater than 10 cm³ showed higher precocity in both cultivation systems, with hydroponic cultivation being more precocious and productive when compared to conventional cultivation.(AU)

Atualmente, um importante movimento econômico está envolvido na aprimoração das bandejas de polietileno para a produção de mudas, sendo desenvolvido com diferentes volumes, números e formatos de células de forma específica para cada cultura. O objetivo do presente estudo foi avaliar o desempenho de alface crespa, oriunda de mudas de bandejas com diferentes volumes, através da comparação entre cultivo convencional e hidropônico-NFT. Cinco volumes de bandejas (10, 20, 30, 40 e 50 cm³ de célula-1) foram testados em cultivo convencional e hidropônico, no período de fevereiro a março de 2016. Para fins de comparação, as plantas foram avaliadas através das características de altura, circunferência, diâmetro do caule, número de folhas, massa fresca da parte aérea e produtividade. O uso de volumes superiores a 10 cm³ apresentou maior precocidade em ambos os sistemas de cultivo, sendo o cultivo hidropônico mais precoce e produtivo quando comparado ao cultivo convencional.(AU)

Senecavirus A (SVA) in finishing swine: diagnosis and viral isolation / Senecavirus A (SVA) em suínos de terminação: diagnóstico e isolamento viral

Senecavirus A (SVA) has been a problem in Brazil since the end of 2014. The infections caused by SVA have disrupted the productive chain in Brazil, as it can be confused with foot-and-mouth disease. Although, the virus has remained endemic in the country, an increase in the number of cases of the disease was observed in 2018. The aim of the present study was to conduct the differential diagnosis of foot-and-mouth disease in an outbreak of vesicular disease in finishing swine. Animals (160-170 days old) were kept on a farm with 6000 pigs in Minas Gerais State, Brazil. The morbidity and mortality rates were 20% and 2.2%, respectively. The diagnosis was performed by RT-PCR, using primers that determine the amplification of an internal region of the 3D gene. Furthermore, samples were inoculated into BHK-21 cell culture for viral isolation. In the first passage under cultivation, a cytopathogenic effect compatible with SVA replication (rounding and detachment of the cell monolayer) was observed. The viral identity was confirmed using two additional assays: indirect immunofluorescence assay (IFA) and nucleotide sequencing. Both tests confirmed that the infection was caused by SVA. In summary, we described a method for the diagnosis and viral isolation of SVA, a virus that arrived in Brazil in 2014 and has become endemic in the country.(AU)

Senecavírus A (SVA) é um problema no Brasil desde o final de 2014. As infecções causadas pelo SVA têm causado problemas para a cadeia produtiva no Brasil, pois podem ser confundidas com febre aftosa. Embora o vírus permaneça endêmico no país, foi observado um aumento no número de casos em 2018. O objetivo do presente estudo foi realizar o diagnostico diferencial de febre aftosa em um surto de doença vesicular em suínos de terminação. Os animais (160 a 170 dias de idade) eram mantidos em uma granja com 6.000 suínos no estado de Minas Gerais, Brasil. As taxas de morbidade e mortalidade foram de 20% e 2,2%, respectivamente. O diagnostico foi realizado por RT-PCR, utilizando primers que determinam a amplificação de uma região interna do gene 3D. Além disso, as amostras foram inoculadas na cultura de células BHK-21 para isolamento viral. Na primeira passagem em cultivo, foi observado efeito citopatogênico compatível com a replicação do SVA (arredondamento e descolamento da monocamada celular). A identidade viral foi confirmada usando duas técnicas adicionais: ensaio de imunofluorescência indireta (IFA) e sequenciamento de nucleotídeos. Ambos os testes confirmaram que a infecção foi causada por SVA. Em resumo, descrevemos um método para o diagnóstico e isolamento viral do SVA, um vírus que chegou ao Brasil em 2014 e se tornou endêmico no país.(AU)

Epigallocatechin Gallate (EGCG) Inhibited the Alv-J-Induced Apoptosis in Df-1 Cells by Inactivation of Nuclear Factor b Pathway

Avian leukosis virus subgroup J (ALV-J), a member of the retroviridae family, can infect both broilers and layers and induce a spectrum of different neoplasms, resulting in serious economic losses in poultry production. Epigallocatechin-3-gallate (EGCG), the major constituent of green tea, has demonstrated remarkable anti-inflammatory and cancer chemopreventive effects in many animal tumor bioassays, cell culture systems and epidemiological studies. To assess the antiviral effects of EGCG on ALV-J-induced cell apoptosis in vitro, DF-1 cells were treated with different EGCG concentrations (0, 5, 10, 20 and 40 µg/mL), and their antiviral effects were examined at different time points (0, 24, 48, 72 and 96 h) using a variety of assays. EGCG alleviated the ALV-J-induced apoptosis in a dose-dependent manner. Because high concentrations (20 and 40 µg/mL) inhibited DF-1 cell growth, and low concentration (5 µg/mL) did not suppress the ALV-J virus, 10 µg/mL was the most appropriate concentration. After 96 h of incubation, 10 µg/mL EGCG improved the ALV-J-triggered suppression of the nuclear transcription factor system by enhancing cytoplasmic NF-B p50/p65 expression and inhibiting nuclear NF-B p50/p65 expression, resulting in decreased cell apoptosis. These results demonstrated that EGCG inhibited ALV-J-induced apoptosis in DF-1 cells in a dose-dependent manner via the NF-B signaling pathway, and that 10 µg/mL EGCG is the optimal concentration, which may be useful for therapeutic drug design.(AU)

Epigallocatechin Gallate (EGCG) Inhibited the Alv-J-Induced Apoptosis in Df-1 Cells by Inactivation of Nuclear Factor b Pathway

Avian leukosis virus subgroup J (ALV-J), a member of the retroviridae family, can infect both broilers and layers and induce a spectrum of different neoplasms, resulting in serious economic losses in poultry production. Epigallocatechin-3-gallate (EGCG), the major constituent of green tea, has demonstrated remarkable anti-inflammatory and cancer chemopreventive effects in many animal tumor bioassays, cell culture systems and epidemiological studies. To assess the antiviral effects of EGCG on ALV-J-induced cell apoptosis in vitro, DF-1 cells were treated with different EGCG concentrations (0, 5, 10, 20 and 40 µg/mL), and their antiviral effects were examined at different time points (0, 24, 48, 72 and 96 h) using a variety of assays. EGCG alleviated the ALV-J-induced apoptosis in a dose-dependent manner. Because high concentrations (20 and 40 µg/mL) inhibited DF-1 cell growth, and low concentration (5 µg/mL) did not suppress the ALV-J virus, 10 µg/mL was the most appropriate concentration. After 96 h of incubation, 10 µg/mL EGCG improved the ALV-J-triggered suppression of the nuclear transcription factor system by enhancing cytoplasmic NF-B p50/p65 expression and inhibiting nuclear NF-B p50/p65 expression, resulting in decreased cell apoptosis. These results demonstrated that EGCG inhibited ALV-J-induced apoptosis in DF-1 cells in a dose-dependent manner via the NF-B signaling pathway, and that 10 µg/mL EGCG is the optimal concentration, which may be useful for therapeutic drug design.

Clinical and pathological findings of Rhodococcus equi infection in foals / Achados clínicos e patológicos da infecção por Rhodococcus equi em potros

Infection by Rhodococcus equi is considered one of the major health concerns for foals worldwide. In order to better understand the disease's clinical and pathological features, we studied twenty cases of natural infection by R. equi in foals. These cases are characterized according to their clinical and pathological findings and immunohistochemical aspects. Necropsy, histologic examination, bacterial culture, R. equi and Pneumocystis spp. immunohistochemistry were performed. The foals had a mean age of 60 days and presented respiratory signs (11/20), hyperthermia (10/20), articular swelling (6/20), prostration (4/20), locomotor impairment (3/20) and diarrhea (3/20), among others. The main lesions were of pyogranulomatous pneumonia, seen in 19 foals, accompanied or not by pyogranulomatous lymphadenitis (10/20) and pyogranulomatous and ulcerative enterocolitis (5/20). Pyogranulomatous osteomyelitis was seen in 3 foals, one of which did not have pulmonary involvement. There was lymphoplasmacytic (4/20), lymphoplasmacytic and neutrophilic (1/20) or pyogranulomatous arthritis (1/20), affecting multiple or singular joints. Immunohistochemistry revealed to be a valuable tool for the detection of R. equi, confirming the diagnosis in all cases. Furthermore, pulmonary immunostaining for Pneumocystis spp. demonstrates that a coinfection with R. equi and this fungal agent is a common event in foals, seen in 13 cases.(AU)

Infecção por Rhodococcus equi é considerado um dos maiores problemas sanitários para potros em todo o mundo. Para melhor compreender a apresentação clínica e patológica da enfermidade, foram avaliados vinte casos de infecção natural por R. equi em potros. Os casos são caracterizados de acordo com seus achados clínicos e patológicos e aspectos imuno-histoquímicos. Foram realizados exames de necropsia, histologia, bacteriologia e imuno-histoquímica para R. equi e Pneumocystis spp. Os potros tinham idade media de 60 dias e apresentaram sinais respiratórios (11/20), hipertermia (10/20), aumento de volume articular (6/20), prostração (4/20), distúrbios locomotores (3/20) e diarreia (3/20), entre outros. As lesões mais importantes eram pneumonia piogranulomatosa, vista em 19 potros, acompanhada ou não por linfadenite piogranulomatosa (10/20) e enterocolite ulcerativa (5/20). Osteomielite piogranulomatosa foi constatada em três potros, um dos quais não apresentava envolvimento pulmonar. Artrites afetando uma ou múltiplas articulações eram caracterizadas por infiltrado linfoplasmocítico (4/20), linfoplasmocítico e neutrofílico (1/20) e piogranulomatoso (1/20). A imuno-histoquímica demonstrou ser uma ferramenta valiosa na detecção de R. equi, permitindo confirmar o diagnóstico em todos os casos avaliados. Além disso, a imuno-histoquímica para Pneumocystis spp. demonstra que a coinfecção por R. equi e o agente fúngico é um evento frequente em potros, constatado em 13 casos.(AU)

Clinical and pathological findings of Rhodococcus equi infection in foals / Achados clínicos e patológicos da infecção por Rhodococcus equi em potros

Infection by Rhodococcus equi is considered one of the major health concerns for foals worldwide. In order to better understand the disease's clinical and pathological features, we studied twenty cases of natural infection by R. equi in foals. These cases are characterized according to their clinical and pathological findings and immunohistochemical aspects. Necropsy, histologic examination, bacterial culture, R. equi and Pneumocystis spp. immunohistochemistry were performed. The foals had a mean age of 60 days and presented respiratory signs (11/20), hyperthermia (10/20), articular swelling (6/20), prostration (4/20), locomotor impairment (3/20) and diarrhea (3/20), among others. The main lesions were of pyogranulomatous pneumonia, seen in 19 foals, accompanied or not by pyogranulomatous lymphadenitis (10/20) and pyogranulomatous and ulcerative enterocolitis (5/20). Pyogranulomatous osteomyelitis was seen in 3 foals, one of which did not have pulmonary involvement. There was lymphoplasmacytic (4/20), lymphoplasmacytic and neutrophilic (1/20) or pyogranulomatous arthritis (1/20), affecting multiple or singular joints. Immunohistochemistry revealed to be a valuable tool for the detection of R. equi, confirming the diagnosis in all cases. Furthermore, pulmonary immunostaining for Pneumocystis spp. demonstrates that a coinfection with R. equi and this fungal agent is a common event in foals, seen in 13 cases.(AU)

Infecção por Rhodococcus equi é considerado um dos maiores problemas sanitários para potros em todo o mundo. Para melhor compreender a apresentação clínica e patológica da enfermidade, foram avaliados vinte casos de infecção natural por R. equi em potros. Os casos são caracterizados de acordo com seus achados clínicos e patológicos e aspectos imuno-histoquímicos. Foram realizados exames de necropsia, histologia, bacteriologia e imuno-histoquímica para R. equi e Pneumocystis spp. Os potros tinham idade media de 60 dias e apresentaram sinais respiratórios (11/20), hipertermia (10/20), aumento de volume articular (6/20), prostração (4/20), distúrbios locomotores (3/20) e diarreia (3/20), entre outros. As lesões mais importantes eram pneumonia piogranulomatosa, vista em 19 potros, acompanhada ou não por linfadenite piogranulomatosa (10/20) e enterocolite ulcerativa (5/20). Osteomielite piogranulomatosa foi constatada em três potros, um dos quais não apresentava envolvimento pulmonar. Artrites afetando uma ou múltiplas articulações eram caracterizadas por infiltrado linfoplasmocítico (4/20), linfoplasmocítico e neutrofílico (1/20) e piogranulomatoso (1/20). A imuno-histoquímica demonstrou ser uma ferramenta valiosa na detecção de R. equi, permitindo confirmar o diagnóstico em todos os casos avaliados. Além disso, a imuno-histoquímica para Pneumocystis spp. demonstra que a coinfecção por R. equi e o agente fúngico é um evento frequente em potros, constatado em 13 casos.(AU)

Enhanced single-cell oil production by cold shock in cyanobacterial cultures / Aumento da produção de óleos unicelulares através de choques de frio em culturas de cianobactérias

The aim of this research was to evaluate the enhanced single-cell oil production by cold shock in Aphanothece microscopica Nägeli using dairy processing wastewater as culture medium. The study focused on (i) temperature optimization for biomass production, (ii) cold shock application to induce lipids biosynthesis and (iii) determination of fatty acids profile under different conditions. Results indicated that temperature of 20°C was the best condition in terms of kinetics parameter, reaching biomass productivities of 160.25mg/L.h. Under these conditions, a lipid content of 12.65% was also observed, resulting in a lipid productivity of 20.27mg/L.h. Additionally, the 0°C cold shock was the most efficient in increasing intracellular lipid content, reaching 28.4% in dry weight. Cold shocks also showed influence on the saturation of fatty acid composition, where the saturated fatty acids decreased, and the monounsaturated and polyunsaturated fatty acids increased by increasing the cold application. Thus, the use of cold shocks indicates to be a key condition for improving the prospects of efficient single-cell oils production.(AU)

O objetivo deste trabalho foi avaliar o aumento da produção de óleos unicelulares por Aphanothece microscopica Nägeli utilizando água residuária de processamento de laticínios como meio de cultura. O estudo concentrou-se na (i) otimização da temperatura para produção de biomassa, (ii) aplicação de choque frio para induzir a biossíntese de lipídios e (iii) determinação do perfil de ácidos graxos sob diferentes condições. Os resultados indicam que a temperatura de 20°C foi a melhor condição em termos de parâmetros cinéticos, atingindo produtividades de biomassa de 160,25mg/L.h. Nesta condição também foi observado um teor lipídico de 12,65%, o que resultou em uma produtividade lipídica de 20,27mg/L.h. Além disso, o choque frio de 0°C foi o mais eficiente para aumentar o conteúdo lipídico intracelular, que atingiu 28,4% em peso seco. Os choques frios também mostraram influência na saturação da composição de ácidos graxos, em que os ácidos graxos saturados diminuíram, e os ácidos graxos monoinsaturados e poli-insaturados ampliaram com o aumento da aplicação de frio. Assim, o uso de choques frios indica ser uma condição chave para melhorar as perspectivas de produção eficiente de óleos unicelulares.(AU)

EFFECT OF SOMATOTROPIN AND THYROXINE ON THE IN VITRO DEVELOPMENT OF BOVINE PREANTRAL FOLLICLES

Abstract The aim of the study was to evaluate the effect of recombinant bovine somatotropin (rbST) and thyroxine (T4) on survival and growth of bovine preantral ovarian follicles (PAOF) cultured in vitro. Ovarian fragments were collected in local abattoirs and immediately fixed for classical histology and transmission electron microscopy (non-cultured control). The other fragments were then cultured in situ for seven days in minimum essential medium alone (MEM+ - cultured control) or in the presence of 1,000 ng/mL rbST and 20 ng/mL T4, isolated or associated. After seven days, there was a reduction (P 0.05) in the percentage of normal follicles in MEM+ alone or with T4. In oocyte diameter, there was a reduction in MEM+ alone. There was no influence (P>0.01) of the medium used on the follicular diameter of the PAOF cultured for seven days. Ultrastructural analysis showed cell damage. In conclusion, the presence of rbST maintains the rate of morphologically normal follicles during the culture for seven days (observed by optical microscopy), but it does not exert beneficial effects on its ultrastructural integrity and oocyte and follicular growth.

Resumo O objetivo do trabalho foi avaliar o efeito da somatotropina recombinante bovina (rbST) e da tiroxina (T4) sobre a sobrevivência e o crescimento de folículos ovarianos pré-antrais (FOPA) bovinos cultivados in vitro. Fragmentos ovarianos foram coletados em abatedouros locais e imediatamente fixados para histologia clássica e microscopia eletrônica de transmissão (controle não cultivado). Os demais fragmentos foram cultivados in situ por sete dias em meio essencial mínimo sozinho (MEM+ - controle cultivado), ou na presença de 1.000 ng/mL de rbST e 20 ng/mL de T4, isoladamente ou em associação. Após sete dias, houve redução (P 0,05) na percentagem de folículos normais no MEM+ sozinho ou quando adicionado T4. No diâmetro oocitário, houve redução no MEM+ sozinho. Não houve influência (P>0,01) do meio utilizado, no diâmetro folicular, dos FOPA cultivados por sete dias. A análise ultraestrutural demonstrou danos celulares. Conclui-se que, apesar da presença da rbST manter a taxa de folículos morfologicamente normais durante o cultivo por sete dias (observado por microscopia óptica), o uso de rbST e T4 em meio de cultivo in vitro de FOPA, nas concentrações utilizadas no presente experimento, não exerce efeitos benéficos quanto à integridade ultraestrutural e aos crescimentos oocitário e folicular.

Epidemiological, clinical and pathological features of canine parvovirus 2c infection in dogs from southern Brazil

ABSTRACT: Canine parvovirus type 2c (CPV-2c) emerged in Europe in the early 2000s and rapidly spread out worldwide. Clinical and molecular data have demonstrated its circulation in Brazilian dogs, yet detailed descriptions of cases are still lacking. This article describes the epidemiological, clinical and pathological features of 24 cases of CPV-2c-associated disease in dogs submitted to veterinary clinics and laboratory diagnosis in southern Brazil (2014-2016). Most affected dogs presented signs/lesions suggestive of parvovirus enteritis: diarrhea, vomiting, hyperemia and hemorrhage of the serous membrane of the small intestine, diffuse segmental granulation, atrophy of the villi, necrosis and fusion of crypts, squamous metaplasia and epithelial syncytia. A number of cases presented features divergent from the classical presentations, including a wide variation in the color of feces (reddish and/or yellowish, light-brownish, orange-brown and brownish), involvement of adults (4/24) and vaccinated dogs (12/24), extensive involvement of the small intestine (8/20) and the presence of pulmonary edema (7/24) and convulsions (3/24). Feces and intestinal fragments submitted to PCR for the CPV-2 VP2 gene and to virus isolation in cell culture yielded positive results in 100% and 58.3% (14/24) of the cases, respectively. Nucleotide sequencing revealed a high nucleotide identity in VP2 (99.4 to 100%) and a consistent mutation at amino acid 426 (asparagine to glutamic acid), considered a signature of CPV-2c. These results confirm the involvement of CPV-2c in the described cases and demonstrate the importance of CPV-2c infection among Brazilian dogs, calling attention of veterinarians to correctly diagnose the disease, mainly considering the frequent atypical presentations.

RESUMO: O parvovírus canino tipo 2c (CPV-2c) surgiu na Europa no início do ano 2000 e rapidamente se espalhou pelas populações de cães ao redor do mundo. Dados clínicos e moleculares demonstraram a sua circulação em cães brasileiros, porém descrições detalhadas desses casos ainda são escassas. Este artigo descreve os aspectos epidemiológicos, clínicos e patológicos de 24 casos de doença gastroentérica associada com a infecção pelo CPV-2c em cães atendidos em clínicas veterinárias e submetidos ao diagnóstico laboratorial no Sul do Brasil (2014-2016). A maioria dos cães afetados apresentaram sinais e/ou lesões sugestivas de enterite por parvovírus: diarreia, vômitos, hiperemia e hemorragia na membrana serosa do intestino delgado, granulação segmentar difusa, atrofia das vilosidades, necrose e fusão de criptas, metaplasia escamosa e sincícios epiteliais. Alguns casos apresentaram características divergentes das apresentações clássicas, incluindo uma grande variação na cor das fezes (avermelhada e/ou amarelada, marrom-claro, marrom-alaranjada ou amarronzada), a participação dos adultos (4/24) e cães vacinados (12/24), um amplo envolvimento do intestino delgado (8/20), a presença de edema pulmonar (7/24) e convulsões (3/24). As fezes e fragmentos intestinais foram submetidos ao teste de PCR para o gene VP2 do CPV-2, e ao isolamento do vírus em cultura de células produziram resultados positivos em 100% e 58,3% (14/24) dos casos, respectivamente. O sequenciamento dos nucleótidos revelou uma alta identidade de nucleótidos na VP2 (99,4-100%) e uma mutação no aminoácido 426 (asparagina para ácido glutâmico), considerada uma assinatura de CPV-2c. Estes resultados confirmam o envolvimento do CPV-2c nos casos descritos e demonstra a importância da infecção pelo CPV-2c entre os cães do Brasil, chamando a atenção de veterinários para diagnosticar corretamente a doença, principalmente considerando-se as apresentações atípicas frequentes.

Bacteriocin-like inhibitory substance in aquaculture: a classic method of protein precipitation for a new aplicability / Substância inibidora tipo bacteriocina na aquicultura: método clássico de precipitação de proteínas para novas aplicações

Techniques to decrease losses from bacterial diseases are always important to improve the fish production. The use of antagonistic substances (bacteriocins) has been proven to be a viable option. The aim of this study was to evaluate different methods of purification for bacteriocin like inhibitory substances (BLIS). For the purification process, we isolated and used two Gram-positive bacilli that produce antagonistic substances for pathogens in aquaculture. Tests for detection of interfering factors were also performed. After the confirmation that the antagonistic action was due the BLIS activity, we carried out the purification methods. The methods tested were: cell free supernatant, acid extraction and ammonium sulfate precipitation at two concentrations (20 and 50%). Salmonella Tiphy CFP/IAL1472 and Aeromonas hydrophila (isolated in a tilapia production environment) were used as indicators of the efficiency of extracts in controlling pathogenic potentials. Ammonium sulfate precipitation at 50% was the most appropriate for purifying the antagonistic substance for both indicators. The extracts of the two isolates remained active for 22 days at 25ºC. These are promising results regarding the water and fish health without the use of antibiotics, in this manner being a safer environmental practice.

Técnicas para diminuir as perdas causadas por doenças bacterianas são importantes para melhorar continuamente a produção de pescado. O uso de substâncias antagônicas (bacteriocinas) tem-se mostrado uma opção viável. O objetivo do trabalho foi avaliar diferentes métodos de purificação de bacteriocinas como substâncias inibidoras (BLIS). Dois bacilos Gram-positivos, produtores de substâncias antagonistas para agentes patogênicos da aquicultura, foram utilizados em processos de purificação. Depois de confirmada a ação antagônica pela atividade de BLIS, os métodos de purificação foram realizados. Os métodos testados foram: células livres de sobrenadante, extração ácida e precipitação por sulfato de amônia em duas concentrações (20 e 50%). Salmonella Typhi PCP/IAL1472 e Aeromonas hydrophila (isolada de um ambiente de tilapicultura) foram utilizadas como indicadores de eficiência dos extratos. O precipitado por sulfato de amônio a 50% foi o mais adequado para purificar a substância antagonista para ambos os isolados indicadores. Os extratos dos dois isolados permaneceram ativos por 22 dias em 25ºC. Estes resultados são promissores do ponto de vista da manutenção da sanidade da água e dos peixes, sem uso de antibióticos, constituindo uma prática ambientalmente mais segura.

Bacteriocin-like inhibitory substance in aquaculture: a classic method of protein precipitation for a new aplicability / Substância inibidora tipo bacteriocina na aquicultura: método clássico de precipitação de proteínas para novas aplicações

Techniques to decrease losses from bacterial diseases are always important to improve the fish production. The use of antagonistic substances (bacteriocins) has been proven to be a viable option. The aim of this study was to evaluate different methods of purification for bacteriocin like inhibitory substances (BLIS). For the purification process, we isolated and used two Gram-positive bacilli that produce antagonistic substances for pathogens in aquaculture. Tests for detection of interfering factors were also performed. After the confirmation that the antagonistic action was due the BLIS activity, we carried out the purification methods. The methods tested were: cell free supernatant, acid extraction and ammonium sulfate precipitation at two concentrations (20 and 50%). Salmonella Tiphy CFP/IAL1472 and Aeromonas hydrophila (isolated in a tilapia production environment) were used as indicators of the efficiency of extracts in controlling pathogenic potentials. Ammonium sulfate precipitation at 50% was the most appropriate for purifying the antagonistic substance for both indicators. The extracts of the two isolates remained active for 22 days at 25ºC. These are promising results regarding the water and fish health without the use of antibiotics, in this manner being a safer environmental practice.(AU)

Técnicas para diminuir as perdas causadas por doenças bacterianas são importantes para melhorar continuamente a produção de pescado. O uso de substâncias antagônicas (bacteriocinas) tem-se mostrado uma opção viável. O objetivo do trabalho foi avaliar diferentes métodos de purificação de bacteriocinas como substâncias inibidoras (BLIS). Dois bacilos Gram-positivos, produtores de substâncias antagonistas para agentes patogênicos da aquicultura, foram utilizados em processos de purificação. Depois de confirmada a ação antagônica pela atividade de BLIS, os métodos de purificação foram realizados. Os métodos testados foram: células livres de sobrenadante, extração ácida e precipitação por sulfato de amônia em duas concentrações (20 e 50%). Salmonella Typhi PCP/IAL1472 e Aeromonas hydrophila (isolada de um ambiente de tilapicultura) foram utilizadas como indicadores de eficiência dos extratos. O precipitado por sulfato de amônio a 50% foi o mais adequado para purificar a substância antagonista para ambos os isolados indicadores. Os extratos dos dois isolados permaneceram ativos por 22 dias em 25ºC. Estes resultados são promissores do ponto de vista da manutenção da sanidade da água e dos peixes, sem uso de antibióticos, constituindo uma prática ambientalmente mais segura.(AU)

Effect of somatotropin and thyroxine on the in vitro development of bovine preantral follicles / Efeito da somatotropina e tiroxina no desenvolvimento in vitro de folículos pré-antrais bovinos

The aim of the study was to evaluate the effect of recombinant bovine somatotropin (rbST) and thyroxine (T4) on survival and growth of bovine preantral ovarian follicles (PAOF) cultured in vitro. Ovarian fragments were collected in local abattoirs and immediately fixed for classical histology and transmission electron microscopy (non-cultured control). The other fragments were then cultured in situ for seven days in minimum essential medium alone (MEM+ - cultured control) or in the presence of 1,000 ng/mL rbST and 20 ng/mL T4, isolated or associated. After seven days, there was a reduction (P0.01) of the medium used on the follicular diameter of the PAOF cultured for seven days. Ultrastructural analysis showed cell damage. In conclusion, the presence of rbST maintains the rate of morphologically normal follicles during the culture for seven days (observed by optical microscopy), but it does not exert beneficial effects on its ultrastructural integrity and oocyte and follicular growth.

O objetivo do trabalho foi avaliar o efeito da somatotropina recombinante bovina (rbST) e da tiroxina (T4) sobre a sobrevivência e o crescimento de folículos ovarianos pré-antrais (FOPA) bovinos cultivados in vitro. Fragmentos ovarianos foram coletados em abatedouros locais e imediatamente fixados para histologia clássica e microscopia eletrônica de transmissão (controle não cultivado). Os demais fragmentos foram cultivados in situ por sete dias em meio essencial mínimo sozinho (MEM+ - controle cultivado), ou na presença de 1.000 ng/mL de rbST e 20 ng/mL de T4, isoladamente ou em associação. Após sete dias, houve redução (P0,01) do meio utilizado, no diâmetro folicular, dos FOPA cultivados por sete dias. A análise ultraestrutural demonstrou danos celulares. Conclui-se que, apesar da presença da rbST manter a taxa de folículos morfologicamente normais durante o cultivo por sete dias (observado por microscopia óptica), o uso de rbST e T4 em meio de cultivo in vitro de FOPA, nas concentrações utilizadas no presente experimento, não exerce efeitos benéficos quanto à integridade ultraestrutural e aos crescimentos oocitário e folicular.

Effect of somatotropin and thyroxine on the in vitro development of bovine preantral follicles / Efeito da somatotropina e tiroxina no desenvolvimento in vitro de folículos pré-antrais bovinos

The aim of the study was to evaluate the effect of recombinant bovine somatotropin (rbST) and thyroxine (T4) on survival and growth of bovine preantral ovarian follicles (PAOF) cultured in vitro. Ovarian fragments were collected in local abattoirs and immediately fixed for classical histology and transmission electron microscopy (non-cultured control). The other fragments were then cultured in situ for seven days in minimum essential medium alone (MEM+ - cultured control) or in the presence of 1,000 ng/mL rbST and 20 ng/mL T4, isolated or associated. After seven days, there was a reduction (P<0.05) in the percentage of normal follicles in MEM+ alone or with T4. In oocyte diameter, there was a reduction in MEM+ alone. There was no influence (P>0.01) of the medium used on the follicular diameter of the PAOF cultured for seven days. Ultrastructural analysis showed cell damage. In conclusion, the presence of rbST maintains the rate of morphologically normal follicles during the culture for seven days (observed by optical microscopy), but it does not exert beneficial effects on its ultrastructural integrity and oocyte and follicular growth.(AU)

O objetivo do trabalho foi avaliar o efeito da somatotropina recombinante bovina (rbST) e da tiroxina (T4) sobre a sobrevivência e o crescimento de folículos ovarianos pré-antrais (FOPA) bovinos cultivados in vitro. Fragmentos ovarianos foram coletados em abatedouros locais e imediatamente fixados para histologia clássica e microscopia eletrônica de transmissão (controle não cultivado). Os demais fragmentos foram cultivados in situ por sete dias em meio essencial mínimo sozinho (MEM+ - controle cultivado), ou na presença de 1.000 ng/mL de rbST e 20 ng/mL de T4, isoladamente ou em associação. Após sete dias, houve redução (P<0,05) na percentagem de folículos normais no MEM+ sozinho ou quando adicionado T4. No diâmetro oocitário, houve redução no MEM+ sozinho. Não houve influência (P>0,01) do meio utilizado, no diâmetro folicular, dos FOPA cultivados por sete dias. A análise ultraestrutural demonstrou danos celulares. Conclui-se que, apesar da presença da rbST manter a taxa de folículos morfologicamente normais durante o cultivo por sete dias (observado por microscopia óptica), o uso de rbST e T4 em meio de cultivo in vitro de FOPA, nas concentrações utilizadas no presente experimento, não exerce efeitos benéficos quanto à integridade ultraestrutural e aos crescimentos oocitário e folicular.(AU)