RESUMO Neste estudo, objetivou-se avaliar a qualidade ambiental de duas áreas do complexo estuarino de São Marcos, utilizando-se as alterações branquiais em Sciades herzbergii como biomarcadores de contaminação aquática. Foram realizadas coletas no período de estiagem (setembro/dezembro de 2015) e no período chuvoso (março/junho de 2016). Foram capturados 40 exemplares de S. herzbergii em dois locais distintos do complexo estuarino: A1) Porto do Itaqui Bacanga (área potencialmente impactada); A2) Igarapé do Puca, Alcântara (área controle). Foram registradas as variáveis abióticas: salinidade, temperatura, pH e oxigênio dissolvido. No laboratório, as brânquias de cada espécime foram fixadas em formol a 10% e mantidas em álcool a 70% até a execução da técnica histológica usual. As alterações branquiais identificadas nos peixes da área potencialmente contaminada foram: deslocamento do epitélio (DE), congestão (CO), aneurisma (AN), fusão lamelar (FL), hiperplasia (HI), dilatação capilar (DC), necrose (NE) e proliferação de células epiteliais (PCE). As alterações na região controle foram: deslocamento do epitélio (DE), fusão lamelar (FL), necrose (NE) e proliferação de células epiteliais (PCE). As alterações branquiais foram mais frequentes na região portuária (84%) do que na região controle (16%). Esses resultados indicam que os peixes da área potencialmente contaminada estão apresentando respostas biológicas ao estresse local.

ABSTRACT This study aimed to compare an environmental quality of two areas of the estuarine complex of São Marcos using gills of changes in Sciades herzbergii as biomarkers of aquatic contamination. Samples were made during the dry season (September/ December 2015) and rainy season (March/ June 2016). They were captured 40 specimens of S. herzbergii at two different points of the estuarine complex: A1) Itaqui port (potentially contaminated area); A2) Igarapé do Puca, Alcântara (control area). Abiotic variables were recorded: salinity, temperature, pH and dissolved oxygen. In the laboratory, the gills of each species were fixed in 10% formaldehyde and kept in 70% alcohol until the usual histological technique was performed. The gill alterations identified in the fish of the potentially contaminated area were: epithelial displacement (DE), congestion (CO), aneurysm (AN), lamellar fusion (FL), hyperplasia (HI), capillary dilation (DC), necrosis and epithelial cell proliferation (PCE). The changes in the control region were: epithelial displacement (DE), lamellar fusion (FL), necrosis (NE) and epithelial cell proliferation (PCE). Gill alterations were more frequent in the port region (84%) than in the control region (16%). These results indicate that fish from the potentially contaminated area are presenting biological responses to local stress.

RESUMO O objetivo deste estudo foi avaliar o efeito da suplementação mineral injetável extra de cobre (Cu) e zinco (Zn) sobre a resposta imunológica de vacas Nelore no período pré-parto. Foram avaliadas 60 vacas prenhes, as quais foram divididas em dois tratamentos, por meio da distribuição aleatória em delineamento inteiramente ao acaso. Aos 75 dias antes do parto, as vacas do tratamento testemunha (T) receberam soro fisiológico como placebo e os animais suplementados (S) receberam mineral injetável via subcutânea (75mg de cobre e 250mg de zinco, em dose única). Foram realizadas três coletas de sangue, duas antes da data prevista para o parto (75 e 10 dias) e uma 30 dias após o parto. Os teores de Cu, Zn, ceruloplasmina, imunoglobulinas G (IgG) e M (IgM) foram analisados durante as três coletas. A atividade fagocitária foi avaliada aos 30 dias pós-parto. Os dados foram examinados mediante análise de variância, com o uso do pacote estatístico do SAS, e os dados individuais da atividade fagocitária pelo PROC GLM. Os dados de Cu, Zn, IgG e IgM foram analisados como medidas repetidas no tempo de coleta por meio do PROC MIXED, com o nível de significância de 5%. Os teores de Cu, Zn, IgM, IgG, ceruloplasmina e a atividade fagocitária das vacas não sofreram influência dos tratamentos (P>0,05). O fornecimento de Cu e Zn injetável, nas doses utilizadas, aos 75 dias antes do parto para vacas Nelore, em dietas suficientes, não alterou os teores de Cu, Zn, ceruloplasmina e a resposta imunológica até 30 dias após o parto.

ABSTRACT The aim of this study was to evaluate the effect of extra injectable mineral supplementation of copper (Cu) and zinc (Zn) on the immune response of Nellore cows in pre-partum period. Sixty pregnant cows were randomly distributed in a completely randomized design in two treatments. In the control treatment (T), cows received saline as placebo, and supplemented treatment (S) received mineral injection (75mg copper and 250mg of zinc, single dose) subcutaneously, 75 days prior to parturition. Blood was sampled three times, two before the expected date of parturition (75 and 10 days) and another at 30 days postpartum. Analyses were performed for Cu, Zn, ceruloplasmin, immunoglobulin G (IgG) and M (IgM) in the three periods and the phagocytic activity in the last period (30 days postpartum). The experimental data were subjected to analysis of variance using the statistical package SAS, being that the individual data phagocytic activity were analyzed by PROC GLM, and the Cu, Zn, IgG and IgM were analyzed as repeated measures in the time, using the PROC MIXED, with the significance level of 5%. The Cu, Zn, IgM, IgG, ceruloplasmin and the phagocytic activity of the cows were not affected by treatments (P>0.05). The supply of injectable Cu and Zn, at the doses used, 75 days before parturition to Nellore cows in sufficient diets, did not alter the serum contents of Cu, Zn, ceruloplasmin and the immune response up to 30 days after parturition.

Neste estudo objetivou-se validar dois órgãos (brânquia e fígado) de Hoplias malabaricus para análise de biomarcadores de contaminação aquática na Área de Proteção Ambiental (APA) do Maracanã em São Luís-MA. Exemplares de traíra foram capturados em uma lagoa de policultivo e no Rio Ambude, localizados na APA do Maracanã. A biometria foi realizada em laboratório. Brânquias e o fígado de cada espécime foram fixados em formol a 10% e mantidos em álcool a 70% até a execução da técnica histológica usual. Cortes de aproximadamente 5µm de espessura foram corados em hematoxilina e eosina (HE). Os resultados referentes à biometria dos peixes capturados durante o período de estiagem indicaram que o comprimento total e padrão dos exemplares foram maiores quando comparados com os indivíduos do período chuvoso. As alterações hepáticas identificadas foram: lipidose, necrose, infiltração leucocitária, vacuolização do citoplasma, hemossiderina e centro de melanomacrófagos. Constataram-se as seguintes lesões branquiais: fusão lamelar, aneurisma lamelar, deslocamento e necrose do epitélio, proliferações de células do muco e dilatação capilar. As respostas biológicas de H. malabaricus, registradas em nível branquial e hepático, podem ser caracterizadas como biomarcadores de contaminação aquática. As brânquias mostraram-se como os melhores órgãos para esse tipo de análise na espécie. Esses resultados indicam que os peixes da região apresentam a saúde comprometida por poluentes presentes nos ambientes analisados.

The aim of the present work was to validate two organs (liver and gill) of Hoplias malabaricus for analysis of aquatic contamination biomarkers in the Maracanã Environmental Protected Area (APA) in São Luís-MA. Trahira animals were captured in a pond polyculture and Ambude River located in Maracanã APA. Biometric analysis was performed in the laboratory. Gills and liver of each specimen were fixed in 10% formalin and kept in 70% alcohol to running the usual histological technique. Cuts of approximately 5μm thick were stained with hematoxylin and eosin (HE). The results concerning the biometrics of the fish caught during the dry season indicated that the total and standard length were higher when compared to the rainy season. The liver changes identified were: lipidosis, necrosis, leukocyte infiltration, vacuolization of the cytoplasm, hemosiderin and melanomacrophages centres. We found the following gill lesions: melting lamellar, lamellar aneurysm displacement, necrosis of the epithelium, proliferation of mucous cells and capillary dilation. The biological responses of H. malabaricus, recorded in gill and liver level, can be characterized as biomarkers of aquatic contamination. The gills appeared as the best organs for this kind of analysis in the species. These results indicate that the fish have health problems caused by pollutants present in the analyzed environments.

Foram utilizadas três vacas não lactantes fistuladas no rúmen com o objetivo de determinar os parâmetros de degradação ruminal in situ da matéria seca (MS) e do amido de grãos de sorgo sob diferentes formas de processamento: 1) Seco e moído (2mm); 2) Seco, moído (3mm), reidratado com aspersão de água sobre o material, e imediatamente ensilado por 30 dias; 3) Seco, inteiro, reidratado por três dias em tubo PVC e, em seguida, moído a 5mm; 4) Seco, inteiro, reidratado por três dias em tubo PVC com aplicação de ar comprimido e, em seguida, moído a 5mm; 5) Seco, inteiro, reidratado por três dias em tubo PVC e, em seguida, moído a 5mm, e ensilado por 30 dias; 6) Seco, inteiro, reidratado por três dias em tubo PVC com aplicação de ar comprimido, moído a 5mm e ensilado por 30 dias. Os teores de proteína bruta, amido, e fibra em detergente neutro (% da MS) variaram de 9,8 a 10,8%; 59,5 a 67,5%; e de 16,1 a 19,3%, respectivamente. Os teores de umidade nos grãos que foram reidratados variaram de 35,8 a 43,9% da MS. A taxa e a extensão de degradação do amido e a degradabilidade potencial da MS foram semelhantes (P>0,05) entre tratamentos. Os grãos de sorgo reidratados com aplicação de ar comprimido, moídos e ensilados (tratamento 6) apresentaram degradabilidade efetiva (DE) da MS e DE do amido sob taxa de passagem ruminal de 2%/h, semelhantes (P>0,05) às observadas para o tratamento 5 (sem aplicação de ar comprimido), e superiores (P<0,05) às dos demais processamentos. Sob taxas de passagem no rúmen de 5 e 8%/h, as DE do amido estimadas para os grãos processados no tratamento 6 foram superiores (P<0,05) às obtidas para os demais tratamentos. O processamento de reidratação por três dias com aplicação de ar comprimido e ensilagem por trinta dias aumentou a degradabilidade efetiva da matéria seca e do amido dos grãos de sorgo, e sua utilização pode ser recomendada para animais em níveis intermediário e elevado de desempenho.

Used three non-lactating cows fistulated in the rumen in order to determine the parameters of in situ ruminal degradation of dry matter (DM) and starch grain sorghum under different forms of processing: 1) Dried and ground (2mm); 2) Dry, crushed (3mm), rehydrated with water spray on the material and immediately ensiled for 30 days, 3) Dry, round, rehydrated for three days in PVC pipe and then crushed to 5mm, 4 ) Dry, full-rehydrated for three days in PVC tube with application of compressed air and then ground to 5mm; 5) Dry, full-rehydrated for three days in PVC pipe and then ground to 5mm, and ensiled for 30 days; 6) Dry, full-rehydrated for three days in PVC tube with application of compressed air, ground to 5mm and ensiled for 30 days. The crude protein, starch, and neutral detergent fiber (% of DM) ranged from 9.8 to 10.8%, 59.5 to 67.5%, and 16.1 to 19.3%, respectively. The moisture in the grains that were rehydrated ranged from 35.8 to 43.9% DM. The rate and extent of starch degradation and the degradability of MS were similar (P>0.05) between treatments. The grain sorghum rehydrated with application of compressed air, ground and ensiled (treatment 6), showed effective degradability (ED) of DM, and DE starch under ruminal passage rate of 2%/h, similar (P>0.05 ) those observed for treating 5 (without application of compressed air) and higher (P<0.05) to the other processing. Under rumen passage rates of 5 and 8%/h, the estimated NDs starch for processed grains in treatment 6 were higher (P<0.05) than those obtained for the other treatments. Processing rehydration for three days with compressed air application and silage for thirty days increased the effective degradability of dry matter and starch grain sorghum, and its use can be recommended for animals in intermediate and high performance.

Paracoccidioidomycosis (PCM) is a chronic systemic mycosis caused by the inhalation of the thermally dimorphic fungus Paracoccidioides brasiliensis as well as the recently described P. lutzii. Because the primary infection occurs in the lungs, we investigated the differential involvement of the right and left lungs in experimental P. brasiliensis infection. Lungs were collected from C57BL/6 mice at 70 days after intravenous infection with 1×106 yeast cells of a virulent strain of P. brasiliensis (Pb18). The left lung, which in mice is smaller and has fewer lobes than the right lung, yielded increased fungal recovery associated with a predominant interleukin-4 response and diminished synthesis of interferon-γ and nitric oxide compared with the right lung. Our data indicate differential involvement of the right and left lungs during experimental PCM. This knowledge emphasizes the need for an accurate, standardized protocol for tissue collection during studies of experimental P. brasiliensis infection, since experiments using the same lungs favor the collection of comparable data among different mice.

RESUMO O objetivo do presente estudo foi determinar a susceptibilidade dos folículos ovarianos, espermatozoides e embriões caprinos ao Vírus da Artrite Encefalite Caprina (CAEV). Para isto, foram analisados espermatozoides e folículos ovarianos pelas técnicas de imunohistoquímica e microscopia eletrônica de transmissão, antes e após protocolos de infecção in vitro com o CAEV. Foram submetidos à análise ultraestrutural, embriões caprinos produzidos in vivo, oriundos de cabras negativas e positivas para o CAEV. Nas amostras seminais, provenientes de animais tanto com infecção natural quanto dos artificialmente infectados, foi observada imunomarcação positiva dos espermatozoides, assim como alterações degenerativas na sua análise ultraestrutural. Já nas amostras de tecido ovariano, a imunomarcação foi mais discreta e identificada na região do estroma. No tocante à análise ultraestrutural, folículos e embriões se apresentaram íntegros. De acordo com esses resultados, pode-se concluir que os espermatozoides caprinos apresentaramse infectados, assinalando a susceptibilidade dessas células ao vírus, bem como a potencialidade do CAEV ser carreado ao cerne do oócito, originando embriões infectados.

ABSTRACT The aim of this study was to determine the susceptibility of goat ovarian follicles, spermatozoa and embryos to caprine arthritis-encephalitis virus (CAEV). Spermatozoa and ovarian follicles were analyzed, before and after in vitro infection with CAEV, through immunohistochemistry and transmission electron microscopy techniques. Goat embryos, produced in vivo from infected and non-infected goats, were submitted to ultrastructural analysis. Immunohistochemical examination of seminal samples from goats naturally and artificially infected with CAEV revealed viral antigens in spermatozoa, while the ultrastructural analysis showed degenerative changes in these cells. Ovarian tissue samples presented a more discreet immunohistochemical positive reaction situated in the stroma region. Ultrastructural analysis revealed that the embryos and ovarian follicles were intact. These results indicate that the spermatozoa were infected, confirming the susceptibility of these cells to the virus, as well as the potential of CAEV entering the oocyte, giving rise to infected embryos.

Chagas' disease, caused by the protozoan Trypanosoma cruzi, is a major cause of cardiovascular disability in countries where it is endemic. Damage to the heart microvasculature has been proposed to be an important factor in the pathogenesis of heart dysfunction. Endothelin-1 (ET-1) is a potent vasoconstrictor and exerts its effects via specific ET A and ET B receptors. A few studies have suggested a role for ET-1 and its receptors in the pathogenesis of Chagas' disease. We investigated the effects of treatment with bosentan, an ET A/ET B receptor antagonist, on the course of T. cruzi infection (Y strain) in C57Bl/6 mice. Treatment with bosentan (100 mg kg-1 day-1) was given per os starting day 0 after infection until sacrifice. Bosentan significantly increased myocardial inflammation, with no effects on parasitemia. Although the total number of nests was similar, a lower number of intact amastigote nests was found in the heart of bosentan-treated animals. Bosentan failed to affect the infection-associated increase in the cardiac levels of the cytokines IFN-g and TNF-a and the chemokines CCL2/MCP-1, CCL3/MIP-1a and CCL5/RANTES. In vitro, pre-incubation with ET-1 (0.1 µM) 4 h before infection enhanced the uptake of the parasites by peritoneal macrophages, and this effect was abrogated when macrophages were pre-treated with bosentan (1 µM) 15 min before incubation with ET-1. However, ET-1 did not alter killing of intracellular parasites after 48 h of in vitro infection. Our data suggest that bosentan-treated mice have a delay in controlling parasitism which is compensated for exacerbated inflammation. Infection is eventually controlled in these animals and lethality is unchanged, demonstrating that ET-1 plays a minor role in the protection against acute murine T. cruzi infection.

Estudaram-se a ocorrência e os fatores predisponentes à artrite encefalite caprina (CAEV) no Rio Grande do Norte. O questionário epidemiológico aplicado a 42 proprietários de 11 municípios produtores de leite avaliou o grau de instrução escolar do proprietário, a origem dos animais, o tipo de criação, as enfermidades mais freqüentes e o manejo sanitário, em geral, bem como as taxas de morbidade e mortalidade. Prevalência de 11% de infecção pelo CAEV foi observada nos rebanhos, com animais soropositivos em todos os municípios pesquisados. As condições de criação mostraram que há fatores que predispõem à introdução ou disseminação da doença na região estudada.

The prevalence of caprine arthritis-encephalitis (CAE) infection in dairy goat herds from Rio Grande do Norte State, Brazil, was estimated, and the predisposing factors to the infection were identified. An epidemiologic survey was held to collect information on educational level of the owner and origin of the animals, and on sanitation and vaccination practices, diseases, morbidity and mortality rates in the herds. The prevalence of CAE virus infection was 11%, and CAE was always observed in some goats in all studied municipalities.

Host resistance to Trypanosoma cruzi infection is dependent on both natural and acquired immune responses. During the early acute phase of infection in mice, natural killer (NK) cell-derived IFN-<FONT FACE="Symbol">g</font> is involved in controlling intracellular parasite replication, mainly through the induction of nitric oxide biosynthesis by activated macrophages. We have shown that IL-12, a powerful inducer of IFN-<FONT FACE="Symbol">g</font> production by NK cells, is synthesized soon after trypomastigote-macrophage interaction. The role of IL-12 in the control of T. cruzi infection in vivo was determined by treating infected mice with anti-IL-12 monoclonal antibody (mAb) and analyzing both parasitemia and mortality during the acute phase of infection. The anti-IL-12 mAb-treated mice had higher levels of parasitemia and mortality compared to control mice. Also, treatment of infected mice with mAb specific for IFN-<FONT FACE="Symbol">g</font> or TNF-<FONT FACE="Symbol">a</font> inhibited the protective effect of exogenous IL-12. On the other hand, TGF-ß and IL-10 produced by infected macrophages inhibited the induction and effects of IL-12. Therefore, while IL-12, TNF-<FONT FACE="Symbol">a</font> and IFN-<FONT FACE="Symbol">g</font> correlate with resistance to T. cruzi infection, TGF-ß and IL-10 promote susceptibility. These results provide support for a role of innate immunity in the control of T. cruzi infection. In addition to its protective role, IL-12 may also be involved in the modulation of T. cruzi-induced myocarditis, since treatment of infected mice with IL-12 or anti-IL-12 mAb leads to an enhanced or decreased inflammatory infiltrate in the heart, respectively. Understanding the role of the cytokines produced during the acute phase of T. cruzi infection and their involvement in protection and pathogenesis would be essential to devise new vaccines or therapies.

Toxoplasma gondii and Trypanosoma cruzi are intracellular parasites which, as part of their life cycle, induce a potent cell-mediated immunity (CMI) maintained by Th1 lymphocytes and IFN-<FONT FACE="Symbol">g</font>. In both cases, induction of a strong CMI is thought to protect the host against rapid parasite multiplication and consequent pathology and lethality during the acute phase of infection. However, the parasitic infection is not eliminated by the immune system and the vertebrate host serves as a parasite reservoir. In contrast, Leishmania sp, which is a slow growing parasite, appears to evade induction of CMI during early stages of infection as a strategy for surviving in a hostile environment (i.e., inside the macrophages which are their obligatory niche in the vertebrate host). Recent reports show that the initiation of IL-12 synthesis by macrophages during these parasitic infections is a key event in regulating CMI and disease outcome. The studies reviewed here indicate that activation/inhibition of distinct signaling pathways and certain macrophage functions by intracellular protozoa are important events in inducing/modulating the immune response of their vertebrate hosts, allowing parasite and host survival and therefore maintaining parasite life cycles.

Em busca da solução para o serio problema da transmissão transfusional da doença de Chagas em nosso meio, realizamos a avaliação de produtos naturais corn atividade tripanosomicida. Os testes "in vitro" foram realizados incubando-se a 4°C, sangue de camundongos contendo ± 10(6)/ ml formas tripomastigotas de Trypanosoma cruzi corn os extratos vegetais. Após diferentes períodos de incubação, frações sangüíneas foram examinadas em microscopia de fase. Os extratos que apresentaram atividade tripanosomicida ate 48 horas de incubação, foram examinados em microscopia eletrônica corn intuito de verificarmos sua toxicidade contra os elementos normais do sangue. Foram testados 242 extratos obtidos de 81 vegetais a 56 demonstraram atividade.

Considering the serius problem of blood transfusion transmission of Chagas disease in our environment, we realized the evalution of natural products against Chagas diseases. The tests were realized "in vitro" by in incubation at 4ºC mouse blood having ± 10(6)/ml of Trypanosoma cruzi, sample "y" and "Colombina" with the plant extracts. After different times of incubation, blood fractions were examined by phase microscopy. The drugs that showed activity until 48 hours of incubation, were examined by electron microscopy to virify the toxicity to blood elements. The tests were realized with 242 extracts from 81 plants and 56 extracts showed activity against T. cruzi.