Babesiosis, anaplasmosis, and trypanosomosis are relevant diseases, potentially causing morbidity in cattle, leading to economic losses. Borreliosis is import as a potential zoonosis. The objective of this study was to determine, by indirect enzyme-linked immunosorbent assay (ELISA), the frequency of seropositive cattle to Babesia bigemina, B. bovis, Anaplasma marginale, Trypanosoma vivax and Borrelia burgdorferi in cattle from the Northeastern region of Pará, Brazil. Sera samples from 246 female adult cattle from municipalities of Castanhal and São Miguel do Guamá were used. Crude antigens ELISAs were used to detect antibodies to all agents, except to A. marginale, to which an indirect ELISA with recombinant major surface 1a protein (MSP1a) antigen was used. Overall frequencies of seropositive animals were: B. bigemina -99.2%; B. bovis -98.8%; A. marginale -68.3%; T. vivax -93.1% and B. burgdorferi -54.9%. The frequencies of seropositive cattle to B. bovis and B. bigemina suggest a high rate of transmission of these organisms by tick in the studied region, which can be classified as enzootically stable to these hemoprotozoans. The low frequency of seropositive cattle to A. marginale may be attributed to a lower sensitivity of the recombinant antigen ELISA utilized or a distinct rate of inoculation of this rickettsia by ticks, as compared with Babesia sp. transmission. The high frequency of seropositive cattle to T. vivax indicates that this hemoprotozoan is prevalent in herds from the Northeastern region of Pará. The rate of animal that showed homologues antibodies to B. burgdorferi indicates the presence of the tickborne spirochaetal agent in the cattle population in the studied region.

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RESUMO Os objetivos deste trabalho foram diagnosticar mediram, em média, 45,4 x 31,8μm, IM 0,7. Pela impressão de pulmão, os ovos (n=100) mediram, em média, 105,3 x 93,6μm, IM 0,9, e as mórulas (n=100) mediram, em média, 43,1 x 36,5μm, IM 0,8. A utilização

ResumoBorrelia burgdorferi é uma espiroqueta transmitida por carrapatos, de ampla distribuição geográfica, que acomete o homem e animais domésticos, tendo os animais silvestres como reservatórios. O presente estudo teve como objetivo relatar a ocorrência de eqüinos soropositivos para B. burgdorferi, por meio de inquérito soroepidemiológico utilizando o ensaio de imunoadsorção enzimática (ELISA) indireto. Procedeu-se a análise sorológica de 747 soros de eqüinos, onde 431 pertenciam ao município de Três Rios e 316 ao de Vassouras, ambos do estado do Rio de Janeiro. Um total de 212 (28,4%) animais foram positivos ao ensaio. O resultado observado foi similar aos descritos para animais no Brasil e Estados Unidos da América, tendo sido verificado que os animais apresentaram anticorpos homólogos anti-Borrelia burgdorferi stricto sensu, corroborando com a ocorrência de Borrelia sp. em eqüinos no estado do Rio de Janeiro.Palavras-chave: soroepidemiologia, borreliose de Lyme, ELISA indireto. AbstractLyme Borreliosis is a widely distributed spirochetosis hosted by wild animals, which transmitted to domestic animals as well as man by ticks. The present study aims to observe the occurrence level of positive tested equines to Borrelia burgdorferi, on two municipalities of Rio de Janeiro State, Brazil, by seroepidemiologic inquiry using the indirect Enzime-Linked Immunosorbent Assay. A serologic analysis was performed on 747 equines serum specimens, which 431 were from Três Rios and 316 were from Vassouras municipality. For this test, 212 (28.4%) animals were positive. The results observed were similar to those described for animals in Brazil and United States of America, confirming that the animals of the studied municipalities showed anti Borrelia burgdorferi homologs antibodies.

The need for alternatives to animal use in pyrogen testing has been driven by the Three Rs concept. This has resulted in the inclusion of the monocyte activation test (MAT) in the European Pharmacopoeia, 2010. However, some technical and regulatory obstacles must be overcome to ensure the effective implementation of the MAT by the industry, especially for the testing of biological products. The yellow fever (YF) vaccine (17DD-YFV) was chosen for evaluation in this study, in view of: a) the 2016–2018 outbreak of YF in Brazil; b) the increase in demand for 17DD-YFV doses; c) the complex production process with live attenuated virus; d) the presence of possible test interference factors, such as residual process components (e.g. ovalbumin); and e) the need for the investigation of other pyrogens that are not detectable by the methods prescribed in the YF vaccine monograph. The product-specific testing was carried out by using cryopreserved and fresh whole blood, and IL-6 and IL-1β levels were used as the marker readouts. After assessing the applicability of the MAT on a 1:10 dilution of 17DD-YFV, endotoxin and non-endotoxin pyrogens were quantified in spiked batches, by using the lipopolysaccharide and lipoteichoic acid standards, respectively. The quantitative analysis demonstrated the correlation between the MAT and the Limulus amoebocyte lysate (LAL) assays, with respect to the limits of endotoxin recovery in spiked batches and the detection of no pyrogenic contamination in commercial batches of 17DD-YFV. The data demonstrated the applicability of the MAT for 17DD-YFV pyrogen testing, and as an alternative method that can contribute to biological quality control studies.