Molecular characterization of Brazilian FeLV strains in São Luis, Maranhão Brazil.

The feline leukemia virus (FeLV) belongs to the Retroviridae family and Gammaretrovirus genus, and causes a variety of neoplastic and non-neoplastic diseases in domestic cats (Felis catus), such as thymic and multicentric lymphomas, myelodysplastic syndromes, acute myeloid leukemia, aplastic anemia, and immunodeficiency. The aim of the present study was to carry out the molecular characterization of FeLV-positive samples and determine the circulating viral subtype in the city of São Luís, Maranhão, Brazil, as well as identify its phylogenetic relationship and genetic diversity. The FIV Ac/FeLV Ag Test Kit (Alere™) and the commercial immunoenzymatic assay kit (Alere™) were used to detect the positive samples, which were subsequently confirmed by ELISA (ELISA - SNAP® Combo FeLV/FIV). To confirm the presence of proviral DNA, a polymerase chain reaction (PCR) was performed to amplify the target fragments of 450, 235, and 166 bp of the FeLV gag gene. For the detection of FeLV subtypes, nested PCR was performed for FeLV-A, B, and C, with amplification of 2350-, 1072-, 866-, and 1755-bp fragments for the FeLV env gene. The results obtained by nested PCR showed that the four positive samples amplified the A and B subtypes. The C subtype was not amplified. There was an AB combination but no ABC combination. Phylogenetic analysis revealed similarities (78% bootstrap) between the subtype circulating in Brazil and FeLV-AB and with the subtypes of Eastern Asia (Japan) and Southeast Asia (Malaysia), demonstrating that this subtype possesses high genetic variability and a differentiated genotype.

Historical highlights regarding glanders, from 2007 to 2017, in the state of Maranhão, Brazil / Destaques históricos do mormo, de 2007 a 2017, no estado do Maranhão, Brasil

Glanders is an infectious disease of equids caused by Burkholderia mallei, a facultative intracellular non-mobile Gram-negative bacterium that can be transmitted to other animals and humans. This study described glanders cases reported in the state of Maranhão, located within an Amazon-Cerrado transition region in northeastern Brazil, from 2007 to 2017. A database in an electronic spreadsheet (Microsoft Excel 2010) was developed containing information on the number of positive animals according to age, sex, purpose, year and month, municipality and mesoregion of origin of the animal. The descriptive analyzis was performed on data provided by the State Agency for Agricultural and Livestock Protection of the state of Maranhão (AGED), and by the official private laboratory. As the database did not have information data about negative animals, possible risk factors could not be evaluated. Among the total of 62,555 equids were evaluated by means of the complement fixation test (CFT), 59,036 were horses, 2,981 mules and 538 donkeys. Thirty-five samples (0.06%) reacted in the CFT. Five additional samples were by the western blot technique and three of them were positive. All the reactive horses (Equuscaballus) were from rural areas. Results presented here indicate that glanders may be endemic in the state of Maranhão and is a public health concern.

O mormo é uma doença infecciosa de equídeos causada por Burkholderia mallei, uma bactéria Gram-negativa intracelular, não móvel e facultativa que pode ser transmitida a outros animais e humanos. O objetivo deste estudo foi descrever os casos de mormo relatados no estado do Maranhão, em uma região de transição Amazônia-Cerrado do Nordeste do Brasil, no período de 2007 a 2017. Um banco de dados em uma planilha eletrônica (Microsoft Excel 2010) foi desenvolvido contendo informações sobre o número de animais positivos de acordo com a idade, sexo, finalidade, ano, mês, municípios e mesorregiões de origem do animal. A análise descritiva foi realizada sobre os dados fornecidos pela Agência Estadual de Defesa Agropecuária do Estado do Maranhão (AGED), e pelo laboratório privado oficial. Como o banco de dados não possui informações sobre animais negativos, possíveis fatores de risco não puderam ser avaliados. Entre o total de 62.555 equídeos foram avaliados com o teste de fixação do complemento (CFT), 59.036 eram cavalos, 2.981 mulas e 538 jumentos. Trinta e cinco (0,06%) amostras reagiram ao CFT. Cinco amostras adicionais foram testadas pela técnica de western blot e três foram positivas. Todos os animais positivos foram cavalos (Equus caballus) da área rural. Os resultados indicam que o mormo pode ser endêmico no estado do Maranhão e uma preocupação de saúde pública.

Diagnosis and phylogenetic analysis of bovine leukemia virus in dairy cattle in northeastern Brazil.

Enzootic bovine leukosis (EBL) is a chronic viral disease of wide distribution in cattle herds and may take several years for the first manifestation of clinical signs. Most animals do not present clinical signs. However, the economic losses are underestimated due to this disease. Thus, this work aimed to detect and characterize BLV in dairy cattle in the Maranhão state, northeastern Brazil. Blood samples were collected from 176 animals from 8 municipalities in the southeastern state of Maranhão. Bovine blood samples were subjected to DNA extraction and molecular diagnosis using nested PCR assays for BLV, targeting gp51 gene. Positive samples were then sequenced and then subjected to phylogenetic inferences. BLV DNA was detected in 16 cattle (16/176, 9.09%) in 4 municipalities. Phylogenetic analyzes showed that the sequence obtained clustered in a clade containing BLV sequences classified as genotype 6, with a high degree of support. Our data shows BLV occurrence in the Northeast of Brazil and the identification of genotype 6 in this region. These findings contribute to the molecular epidemiology of this agent in Brazil.

High genetic diversity of Anaplasma marginale infecting dairy cattle in northeastern Brazil.

Anaplasma marginale is an obligate intracellular Gram-negative bacterium found in ruminants' erythrocytes and is the etiological agent of bovine anaplasmosis. The bacterium's genetic diversity has been characterized based on sequences of major surface proteins (MSPs), such as MSP1α. The aim of the present study was to investigate the genetic diversity of A. marginale in cattle in the state of Maranhão, northeastern Brazil. To this end, 343 blood samples were harvested and subjected to iELISA assays using the recombinant surface protein MSP5. Out of 343 blood samples, 235 (68.5%) were randomly chosen and submitted to DNA extraction, qPCR and conventional PCR targeting the msp1α gene to determine amino acid sequences and classify the genotypes. The iELISA results showed 81.34% seropositivity (279/343), whereas qPCR revealed 224 positive samples (95.32%). Among these qPCR-positive samples, 67.4% (151/224) were also positive in the cPCR. Among the 50 obtained sequences, 21 strains had not been previously reported. Regarding the genotypes, H (26/50) and E (18/50) were identified most often, while genotypes F and C were only identified twice each and B and G once each. In conclusion, high prevalence and genetic diversity for A. marginale were observed in dairy cattle herds in the state of Maranhão.

Anti-Ehrlichia properties of the dichloromethane fraction of Ageratum conyzoides associated with doxycycline: In vitro study / Propriedades anti-Ehrlichia da fração diclorometânica de Ageratum conyzoides associada com doxiciclina: estudo in vitro

The increasing number of cases of canine ehrlichiosis caused by Ehrlichia canis in hospitals and veterinary clinics has demonstrated the need for a new drug protocol for this disease. Doxycycline is used to treat ehrlichiosis, but the resistance of the microorganism to this treatment protocol, as well as the various side effects to the animals, has become a concern. Several studies have shown a positive interaction with extracts of plants and drugs, which allow for the reduction of the concentration necessary to produce the desired effect, minimizing adverse effects. This study determined the efficiency of the combination of the dichloromethane (DCM) fraction of Ageratum conyzoides L. with anti-Ehrlichia activity and doxycycline by using the checkerboard assay. Plant material was collected in São Luís, northeastern Brazil, followed by extraction in MeOH: H2O (8:2) and partitioning of the DCM fraction. After determining the Minimum Inhibitory Concentration (MIC) of the fraction under study against DH82 cells infected with Ehrlichia canis, it was combined with doxycycline to derive the Fractional Inhibitory Concentration Index (CIF Index). A reduction of 5.83 times the doxycycline minimum inhibitory concentration was observed, showing that this fraction of A. conyzoides composed predominantly by the class of lignans, identified by mass spectrometry notably intensified the activity of doxycycline against E. canis, resulting in a synergistic effect.(AU)

O crescente número de casos de erliquiose canina por Ehrlichia canis em hospitais e clínicas veterinárias tem demonstrado a necessidade de um novo protocolo de medicamentos para essa doença. A doxiciclina é usada para tratar a erliquiose, mas a resistência do microrganismo a esse protocolo de tratamento, bem como os diversos efeitos colaterais para os animais, tornou-se uma preocupação. Vários estudos têm demonstrado interação positiva com extratos de plantas e fármacos, que permitem a redução da concentração necessária para produzir o efeito desejado, minimizando os efeitos adversos. Este estudo determinou a eficiência da combinação da fração diclorometânica (DCM) de Ageratum conyzoides L. com atividade anti-Ehrlichia canis associada com doxiciclina por meio do ensaio de Checkerboard. O material vegetal foi coletado em São Luís, Maranhão, nordeste do Brasil, seguido pela extração em MeOH:H2O (8:2) e partição da fração diclorometânica. Após a determinação da Concentração Inibitória Mínima (CIM) da fração em estudo frente às células DH82 infectadas com Ehrlichia canis, a mesma foi combinada com a doxiciclina para derivação do Índice de Concentração Inibitória da Fração (Índice CIF). Observou-se uma redução de 5,83 vezes a concentração inibitória mínima da doxiciclina mostrando que esta fração de A. conyzoides, composta predominantemente por lignanas identificadas por espectrometria de massas, notavelmente intensificou a atividade desse fármaco contra E. canis, resultando em um efeito sinérgico.(AU)

High genetic diversity of Anaplasma marginale infecting dairy cattle in northeastern Brazil / Alta diversidade genética de Anaplasma marginale em bovinos de leite no nordeste do Brasil

Abstract Anaplasma marginale is an obligate intracellular Gram-negative bacterium found in ruminants' erythrocytes and is the etiological agent of bovine anaplasmosis. The bacterium's genetic diversity has been characterized based on sequences of major surface proteins (MSPs), such as MSP1α. The aim of the present study was to investigate the genetic diversity of A. marginale in cattle in the state of Maranhão, northeastern Brazil. To this end, 343 blood samples were harvested and subjected to iELISA assays using the recombinant surface protein MSP5. Out of 343 blood samples, 235 (68.5%) were randomly chosen and submitted to DNA extraction, qPCR and conventional PCR targeting the msp1α gene to determine amino acid sequences and classify the genotypes. The iELISA results showed 81.34% seropositivity (279/343), whereas qPCR revealed 224 positive samples (95.32%). Among these qPCR-positive samples, 67.4% (151/224) were also positive in the cPCR. Among the 50 obtained sequences, 21 strains had not been previously reported. Regarding the genotypes, H (26/50) and E (18/50) were identified most often, while genotypes F and C were only identified twice each and B and G once each. In conclusion, high prevalence and genetic diversity for A. marginale were observed in dairy cattle herds in the state of Maranhão.

Resumo Anaplasma marginale é uma bactéria Gram-negativa intracelular obrigatória de eritrócitos de ruminantes e responsável pela anaplasmose bovina. A diversidade genética de A. marginale tem sido caracterizada com base nas sequências das principais proteínas de superfície (MSPs), como a MSP1α. O objetivo deste estudo foi investigar a diversidade genética de A. marginale em bovinos no estado do Maranhão, Nordeste do Brasil. Dessa forma, 343 amostras de sangue foram submetidas ao ensaio iELISA, utilizando-se a proteína recombinante MSP5. Das 343 amostras de sangue, 235 (68,5%) foram escolhidas aleatoriamente e submetidas à extração de DNA, qPCR e PCR convencional para gene msp1α, para determinação das sequências de aminoácidos e classificação dos genótipos. Os resultados do iELISA mostraram 81,34% de soropositividade (279/343), enquanto qPCR revelou 224 amostras positivas (95,32%). Dentre estas na qPCR, 67,4% (151/224) mostraram-se positivas no PCR convencional. Das 50 sequências obtidas, 21 cepas não haviam sido relatadas anteriormente. Em relação aos genótipos, H (26/50) e E (18/50) foram os mais frequentes, enquanto os genótipos F e C foram identificados apenas duas vezes cada, e B e G uma vez cada. Em conclusão, alta prevalência e marcante diversidade genética de A. marginale foram observadas em rebanhos leiteiros no estado do Maranhão.

Anti-Ehrlichia properties of the dichloromethane fraction of Ageratum conyzoides associated with doxycycline: In vitro study / Propriedades anti-Ehrlichia da fração diclorometânica de Ageratum conyzoides associada com doxiciclina: estudo in vitro

ABSTRACT: The increasing number of cases of canine ehrlichiosis caused by Ehrlichia canis in hospitals and veterinary clinics has demonstrated the need for a new drug protocol for this disease. Doxycycline is used to treat ehrlichiosis, but the resistance of the microorganism to this treatment protocol, as well as the various side effects to the animals, has become a concern. Several studies have shown a positive interaction with extracts of plants and drugs, which allow for the reduction of the concentration necessary to produce the desired effect, minimizing adverse effects. This study determined the efficiency of the combination of the dichloromethane (DCM) fraction of Ageratum conyzoides L. with anti-Ehrlichia activity and doxycycline by using the checkerboard assay. Plant material was collected in São Luís, northeastern Brazil, followed by extraction in MeOH: H2O (8:2) and partitioning of the DCM fraction. After determining the Minimum Inhibitory Concentration (MIC) of the fraction under study against DH82 cells infected with Ehrlichia canis, it was combined with doxycycline to derive the Fractional Inhibitory Concentration Index (CIF Index). A reduction of 5.83 times the doxycycline minimum inhibitory concentration was observed, showing that this fraction of A. conyzoides composed predominantly by the class of lignans, identified by mass spectrometry notably intensified the activity of doxycycline against E. canis, resulting in a synergistic effect.

RESUMO: O crescente número de casos de erliquiose canina por Ehrlichia canis em hospitais e clínicas veterinárias tem demonstrado a necessidade de um novo protocolo de medicamentos para essa doença. A doxiciclina é usada para tratar a erliquiose, mas a resistência do microrganismo a esse protocolo de tratamento, bem como os diversos efeitos colaterais para os animais, tornou-se uma preocupação. Vários estudos têm demonstrado interação positiva com extratos de plantas e fármacos, que permitem a redução da concentração necessária para produzir o efeito desejado, minimizando os efeitos adversos. Este estudo determinou a eficiência da combinação da fração diclorometânica (DCM) de Ageratum conyzoides L. com atividade anti-Ehrlichia canis associada com doxiciclina por meio do ensaio de Checkerboard. O material vegetal foi coletado em São Luís, Maranhão, nordeste do Brasil, seguido pela extração em MeOH:H2O (8:2) e partição da fração diclorometânica. Após a determinação da Concentração Inibitória Mínima (CIM) da fração em estudo frente às células DH82 infectadas com Ehrlichia canis, a mesma foi combinada com a doxiciclina para derivação do Índice de Concentração Inibitória da Fração (Índice CIF). Observou-se uma redução de 5,83 vezes a concentração inibitória mínima da doxiciclina mostrando que esta fração de A. conyzoides, composta predominantemente por lignanas identificadas por espectrometria de massas, notavelmente intensificou a atividade desse fármaco contra E. canis, resultando em um efeito sinérgico.

Anti-Ehrlichia properties of the dichloromethane fraction of Ageratum conyzoides associated with doxycycline: In vitro study / Propriedades anti-Ehrlichia da fração diclorometânica de Ageratum conyzoides associada com doxiciclina: estudo in vitro

The increasing number of cases of canine ehrlichiosis caused by Ehrlichia canis in hospitals and veterinary clinics has demonstrated the need for a new drug protocol for this disease. Doxycycline is used to treat ehrlichiosis, but the resistance of the microorganism to this treatment protocol, as well as the various side effects to the animals, has become a concern. Several studies have shown a positive interaction with extracts of plants and drugs, which allow for the reduction of the concentration necessary to produce the desired effect, minimizing adverse effects. This study determined the efficiency of the combination of the dichloromethane (DCM) fraction of Ageratum conyzoides L. with anti-Ehrlichia activity and doxycycline by using the checkerboard assay. Plant material was collected in São Luís, northeastern Brazil, followed by extraction in MeOH: H2O (8:2) and partitioning of the DCM fraction. After determining the Minimum Inhibitory Concentration (MIC) of the fraction under study against DH82 cells infected with Ehrlichia canis, it was combined with doxycycline to derive the Fractional Inhibitory Concentration Index (CIF Index). A reduction of 5.83 times the doxycycline minimum inhibitory concentration was observed, showing that this fraction of A. conyzoides composed predominantly by the class of lignans, identified by mass spectrometry notably intensified the activity of doxycycline against E. canis, resulting in a synergistic effect.

O crescente número de casos de erliquiose canina por Ehrlichia canis em hospitais e clínicas veterinárias tem demonstrado a necessidade de um novo protocolo de medicamentos para essa doença. A doxiciclina é usada para tratar a erliquiose, mas a resistência do microrganismo a esse protocolo de tratamento, bem como os diversos efeitos colaterais para os animais, tornou-se uma preocupação. Vários estudos têm demonstrado interação positiva com extratos de plantas e fármacos, que permitem a redução da concentração necessária para produzir o efeito desejado, minimizando os efeitos adversos. Este estudo determinou a eficiência da combinação da fração diclorometânica (DCM) de Ageratum conyzoides L. com atividade anti-Ehrlichia canis associada com doxiciclina por meio do ensaio de Checkerboard. O material vegetal foi coletado em São Luís, Maranhão, nordeste do Brasil, seguido pela extração em MeOH:H2O (8:2) e partição da fração diclorometânica. Após a determinação da Concentração Inibitória Mínima (CIM) da fração em estudo frente às células DH82 infectadas com Ehrlichia canis, a mesma foi combinada com a doxiciclina para derivação do Índice de Concentração Inibitória da Fração (Índice CIF). Observou-se uma redução de 5,83 vezes a concentração inibitória mínima da doxiciclina mostrando que esta fração de A. conyzoides, composta predominantemente por lignanas identificadas por espectrometria de massas, notavelmente intensificou a atividade desse fármaco contra E. canis, resultando em um efeito sinérgico.

Occurrence of Bartonella genotypes in bats and associated Streblidae flies from Maranhão state, northeastern Brazil.

Bartonella is a genus of emerging zoonotic bacteria that are mainly associated with mammalian erythrocytes and endothelial cells. Bats are natural reservoirs for a variety of important pathogens that impact human and animal health. Recent reports have highlighted the role of bats and bat flies in the maintenance of Bartonella. Here, we showed that none of the 29 bat DNA blood samples obtained from five bat species in São Luís Island, state of Maranhão, northeastern Brazil, were positive for Bartonella in qPCR assays targeting nuoG. On the other hand, three out of 15 DNA samples (20%) from flies in the family Streblidae were positive for Bartonella. The BLASTn results showed that the gltA and rpoB sequences shared identities ranging from 97.2% to 100%, with Bartonella sequences amplified from bats or bat flies from Costa Rica and Brazil. These findings were supported by phylogenetic analyses based on Bayesian inferences. The present study showed that Bartonella genotypes are present in bat flies, thus shedding some light on the distribution of bat fly-related Bartonella genotypes in South America.

Occurrence of Bartonella genotypes in bats and associated Streblidae flies from Maranhão state, northeastern Brazil / Ocorrência de genótipos de Bartonella em morcegos e moscas Streblidae no Estado do Maranhão, Nordeste do Brasil

Bartonella is a genus of emerging zoonotic bacteria that are mainly associated with mammalian erythrocytes and endothelial cells. Bats are natural reservoirs for a variety of important pathogens that impact human and animal health. Recent reports have highlighted the role of bats and bat flies in the maintenance of Bartonella. Here, we showed that none of the 29 bat DNA blood samples obtained from five bat species in São Luís Island, state of Maranhão, northeastern Brazil, were positive for Bartonella in qPCR assays targeting nuoG. On the other hand, three out of 15 DNA samples (20%) from flies in the family Streblidae were positive for Bartonella. The BLASTn results showed that the gltA and rpoB sequences shared identities ranging from 97.2% to 100%, with Bartonella sequences amplified from bats or bat flies from Costa Rica and Brazil. These findings were supported by phylogenetic analyses based on Bayesian inferences. The present study showed that Bartonella genotypes are present in bat flies, thus shedding some light on the distribution of bat fly-related Bartonella genotypes in South America.(AU)

Bartonella é um gênero de bactérias zoonóticas emergentes associadas principalmente a eritrócitos e células endoteliais de mamíferos. Morcegos são reservatórios naturais para uma variedade de patógenos importantes que afetam a saúde humana e animal. Além disso, estudos recentes destacaram o papel dos morcegos e de moscas associadas a morcegos na manutenção de Bartonella. No presente estudo, nenhuma das 29 amostras de DNA obtidas a partir do sangue de cinco espécies de morcegos amostrados na ilha de São Luís, estado do Maranhão, Nordeste do Brasil, foi positiva para Bartonella nos ensaios de qPCR direcionados ao gene nuoG. Por outro lado, três das 15 (20%) amostras de DNA de moscas da família Streblidae foram positivas para Bartonella. Os resultados do BLASTn mostraram que as sequências dos genes gltA e rpoB compartilharam identidade, variando de 97,2% a 100%, com as sequências de Bartonella amplificadas em morcegos ou moscas amostrados na Costa Rica ou Brasil. Tais resultados corroboraram as análises filogenéticas realizadas por Inferência Bayesiana. O presente estudo mostrou a ocorrência de Bartonella em moscas de morcegos, auxiliando a esclarecer a distribuição dos genótipos de Bartonella relacionadas a moscas Streblidae na América do Sul.(AU)

Occurrence of Bartonella genotypes in bats and associated Streblidae flies from Maranhão state, northeastern Brazil / Ocorrência de genótipos de Bartonella em morcegos e moscas Streblidae no Estado do Maranhão, Nordeste do Brasil

Abstract Bartonella is a genus of emerging zoonotic bacteria that are mainly associated with mammalian erythrocytes and endothelial cells. Bats are natural reservoirs for a variety of important pathogens that impact human and animal health. Recent reports have highlighted the role of bats and bat flies in the maintenance of Bartonella. Here, we showed that none of the 29 bat DNA blood samples obtained from five bat species in São Luís Island, state of Maranhão, northeastern Brazil, were positive for Bartonella in qPCR assays targeting nuoG. On the other hand, three out of 15 DNA samples (20%) from flies in the family Streblidae were positive for Bartonella. The BLASTn results showed that the gltA and rpoB sequences shared identities ranging from 97.2% to 100%, with Bartonella sequences amplified from bats or bat flies from Costa Rica and Brazil. These findings were supported by phylogenetic analyses based on Bayesian inferences. The present study showed that Bartonella genotypes are present in bat flies, thus shedding some light on the distribution of bat fly-related Bartonella genotypes in South America.

Resumo Bartonella é um gênero de bactérias zoonóticas emergentes associadas principalmente a eritrócitos e células endoteliais de mamíferos. Morcegos são reservatórios naturais para uma variedade de patógenos importantes que afetam a saúde humana e animal. Além disso, estudos recentes destacaram o papel dos morcegos e de moscas associadas a morcegos na manutenção de Bartonella. No presente estudo, nenhuma das 29 amostras de DNA obtidas a partir do sangue de cinco espécies de morcegos amostrados na ilha de São Luís, estado do Maranhão, Nordeste do Brasil, foi positiva para Bartonella nos ensaios de qPCR direcionados ao gene nuoG. Por outro lado, três das 15 (20%) amostras de DNA de moscas da família Streblidae foram positivas para Bartonella. Os resultados do BLASTn mostraram que as sequências dos genes gltA e rpoB compartilharam identidade, variando de 97,2% a 100%, com as sequências de Bartonella amplificadas em morcegos ou moscas amostrados na Costa Rica ou Brasil. Tais resultados corroboraram as análises filogenéticas realizadas por Inferência Bayesiana. O presente estudo mostrou a ocorrência de Bartonella em moscas de morcegos, auxiliando a esclarecer a distribuição dos genótipos de Bartonella relacionadas a moscas Streblidae na América do Sul.

Anti-Ehrlichia properties of the essential oil of Ageratum conyzoides L. and its interaction with doxycycline.

Canine Monocytic Ehrlichiosis (CME) is an infectious disease caused by the rickettsia organism Ehrlichia canis which is transmitted mainly the ixodid brown dog tick Rhipicephalus sanguineus. The prevalence of E. canis infection has been increasing in recent years. The World Health Organization has been warned about antibiotics resistance and one of the way to prevent this situation is found new compound with this property. Doxycycline is the treatment of choice for this tick-borne disease. Adverse effects are noted in dogs that are sensitive to this drug. Antibiotic resistance may also occur. The present study aimed to evaluate the anti-Ehrlichia properties of the essential oil of the aerial parts of Ageratum conyzoides L. in infected DH82 cells, as well as its anti-Ehrlichia activity associated with doxycycline using the checkerboard assay. A. conyzoides is a native plant from northeast Brazil with many reports of ethnopharmacological applications. The essential oil of A. conyzoides was extracted from the aerial parts of the plant using the hydrodistillation method. E. canis-infected DH82 cells were cultured in DMEM (Dulbecco's Modified Eagle Medium), maintained at 37 °C and 5% CO2, and standardized at a 70% infection rate for the initiation of treatment protocols. The tests were first carried out with the aim of defining the IC50. The combined effect of doxycycline and A. conyzoides essential oil was then determined using the checkerboard dilution technique (checkerboard method) in which the IC50 was 200 µg/mL. The doxycycline reduction index from the combined effect was 4.90 times resulting in a synergistic effect. To the authors' knowledge, this is the first alternative treatment (alternative therapy) based on bioactive molecules that have antibacterial activity against E. canis.

Molecular detection of Mycoplasma suis in extensive pig production systems in the State of Maranhão, northeast Brazil / Detecção molecular de Mycoplasma suis em suínos de criações extensivas no estado do Maranhão, nordeste do Brasil

Abstract Mycoplasma suis is a bacterium that causes hemoplasmosis in pigs. This agent is capable of adhering to the surface of porcine erythrocytes, inducing structural changes on these cells. In Brazil, there are few reports about the disease, its causal agent, and the economic impact of this pathogen on pig production systems and farm sanitation. The present study aimed to investigate the occurrence of M. suis in extensive swine farms located in the counties of Itapecuru Mirim, Santa Rita and Rosario, State of Maranhão, northeast Brazil. For such purpose, 64 blood samples of pigs from these facilities were tested for M. suis using a 16S rRNA gene-based quantitative real-time PCR (qPCR); 82.3%, 65.2% and 25% of blood samples of swine from farms in the cities of Itapecuru Mirim, Santa Rita and Rosario were positive for M. suis by qPCR, respectively. This study shows, for the first time, that M. suis circulates in pig populations from the state of Maranhão, Northeast Brazil.

Resumo Mycoplasma suis é uma bactéria que causa a hemoplasmose em suínos. Este agente é capaz de se aderir à superfície dos eritrócitos de suínos, ocasionando deformações estruturais nestas células. No Brasil, poucos são os relatos acerca do parasita, da infecção e de seus impactos econômicos nas esferas produtiva e sanitária. O objetivo deste estudo foi investigar, por meio da PCR em tempo real quantitativa (qPCR) baseada no gene 16S rRNA, a ocorrência de M. suis em 64 amostras de sangue de suínos de criações extensivas dos municípios de Itapecuru Mirim, Santa Rita e Rosário, localizados no estado do Maranhão. Foram obtidos um percentual de 82,3%, 65,2% e 25% de amostras positivas na qPCR para M. suis nos municípios de Itapecuru Mirim, Santa Rita e Rosário, respectivamente. Este estudo mostra que M. suis circula entre os suínos de criações extensivas no estado do Maranhão.

Molecular detection of Mycoplasma suis in extensive pig production systems in the State of Maranhão, northeast Brazil.

Mycoplasma suis is a bacterium that causes hemoplasmosis in pigs. This agent is capable of adhering to the surface of porcine erythrocytes, inducing structural changes on these cells. In Brazil, there are few reports about the disease, its causal agent, and the economic impact of this pathogen on pig production systems and farm sanitation. The present study aimed to investigate the occurrence of M. suis in extensive swine farms located in the counties of Itapecuru Mirim, Santa Rita and Rosario, State of Maranhão, northeast Brazil. For such purpose, 64 blood samples of pigs from these facilities were tested for M. suis using a 16S rRNA gene-based quantitative real-time PCR (qPCR); 82.3%, 65.2% and 25% of blood samples of swine from farms in the cities of Itapecuru Mirim, Santa Rita and Rosario were positive for M. suis by qPCR, respectively. This study shows, for the first time, that M. suis circulates in pig populations from the state of Maranhão, Northeast Brazil.

Molecular detection of Mycoplasma suis in extensive pig production systems in the State of Maranhão, northeast Brazil / Detecção molecular de Mycoplasma suis em suínos de criações extensivas no estado do Maranhão, nordeste do Brasil

Mycoplasma suis is a bacterium that causes hemoplasmosis in pigs. This agent is capable of adhering to the surface of porcine erythrocytes, inducing structural changes on these cells. In Brazil, there are few reports about the disease, its causal agent, and the economic impact of this pathogen on pig production systems and farm sanitation. The present study aimed to investigate the occurrence of M. suis in extensive swine farms located in the counties of Itapecuru Mirim, Santa Rita and Rosario, State of Maranhão, northeast Brazil. For such purpose, 64 blood samples of pigs from these facilities were tested for M. suis using a 16S rRNA gene-based quantitative real-time PCR (qPCR); 82.3%, 65.2% and 25% of blood samples of swine from farms in the cities of Itapecuru Mirim, Santa Rita and Rosario were positive for M. suis by qPCR, respectively. This study shows, for the first time, that M. suis circulates in pig populations from the state of Maranhão, Northeast Brazil.(AU)

Mycoplasma suis é uma bactéria que causa a hemoplasmose em suínos. Este agente é capaz de se aderir à superfície dos eritrócitos de suínos, ocasionando deformações estruturais nestas células. No Brasil, poucos são os relatos acerca do parasita, da infecção e de seus impactos econômicos nas esferas produtiva e sanitária. O objetivo deste estudo foi investigar, por meio da PCR em tempo real quantitativa (qPCR) baseada no gene 16S rRNA, a ocorrência de M. suis em 64 amostras de sangue de suínos de criações extensivas dos municípios de Itapecuru Mirim, Santa Rita e Rosário, localizados no estado do Maranhão. Foram obtidos um percentual de 82,3%, 65,2% e 25% de amostras positivas na qPCR para M. suis nos municípios de Itapecuru Mirim, Santa Rita e Rosário, respectivamente. Este estudo mostra que M. suis circula entre os suínos de criações extensivas no estado do Maranhão.(AU)

Molecular detection of Mycoplasma suis in extensive pig production systems in the State of Maranhão, northeast Brazil / Detecção molecular de Mycoplasma suis em suínos de criações extensivas no estado do Maranhão, nordeste do Brasil

Mycoplasma suis is a bacterium that causes hemoplasmosis in pigs. This agent is capable of adhering to the surface of porcine erythrocytes, inducing structural changes on these cells. In Brazil, there are few reports about the disease, its causal agent, and the economic impact of this pathogen on pig production systems and farm sanitation. The present study aimed to investigate the occurrence of M. suis in extensive swine farms located in the counties of Itapecuru Mirim, Santa Rita and Rosario, State of Maranhão, northeast Brazil. For such purpose, 64 blood samples of pigs from these facilities were tested for M. suis using a 16S rRNA gene-based quantitative real-time PCR (qPCR); 82.3%, 65.2% and 25% of blood samples of swine from farms in the cities of Itapecuru Mirim, Santa Rita and Rosario were positive for M. suis by qPCR, respectively. This study shows, for the first time, that M. suis circulates in pig populations from the state of Maranhão, Northeast Brazil.(AU)

Mycoplasma suis é uma bactéria que causa a hemoplasmose em suínos. Este agente é capaz de se aderir à superfície dos eritrócitos de suínos, ocasionando deformações estruturais nestas células. No Brasil, poucos são os relatos acerca do parasita, da infecção e de seus impactos econômicos nas esferas produtiva e sanitária. O objetivo deste estudo foi investigar, por meio da PCR em tempo real quantitativa (qPCR) baseada no gene 16S rRNA, a ocorrência de M. suis em 64 amostras de sangue de suínos de criações extensivas dos municípios de Itapecuru Mirim, Santa Rita e Rosário, localizados no estado do Maranhão. Foram obtidos um percentual de 82,3%, 65,2% e 25% de amostras positivas na qPCR para M. suis nos municípios de Itapecuru Mirim, Santa Rita e Rosário, respectivamente. Este estudo mostra que M. suis circula entre os suínos de criações extensivas no estado do Maranhão.(AU)

Molecular detection of Mycoplasma suis in extensive pig production systems in the State of Maranhão, northeast Brazil / Detecção molecular de Mycoplasma suis em suínos de criações extensivas no estado do Maranhão, nordeste do Brasil

Mycoplasma suis is a bacterium that causes hemoplasmosis in pigs. This agent is capable of adhering to the surface of porcine erythrocytes, inducing structural changes on these cells. In Brazil, there are few reports about the disease, its causal agent, and the economic impact of this pathogen on pig production systems and farm sanitation. The present study aimed to investigate the occurrence of M. suis in extensive swine farms located in the counties of Itapecuru Mirim, Santa Rita and Rosario, State of Maranhão, northeast Brazil. For such purpose, 64 blood samples of pigs from these facilities were tested for M. suis using a 16S rRNA gene-based quantitative real-time PCR (qPCR); 82.3%, 65.2% and 25% of blood samples of swine from farms in the cities of Itapecuru Mirim, Santa Rita and Rosario were positive for M. suis by qPCR, respectively. This study shows, for the first time, that M. suis circulates in pig populations from the state of Maranhão, Northeast Brazil.

Mycoplasma suis é uma bactéria que causa a hemoplasmose em suínos. Este agente é capaz de se aderir à superfície dos eritrócitos de suínos, ocasionando deformações estruturais nestas células. No Brasil, poucos são os relatos acerca do parasita, da infecção e de seus impactos econômicos nas esferas produtiva e sanitária. O objetivo deste estudo foi investigar, por meio da PCR em tempo real quantitativa (qPCR) baseada no gene 16S rRNA, a ocorrência de M. suis em 64 amostras de sangue de suínos de criações extensivas dos municípios de Itapecuru Mirim, Santa Rita e Rosário, localizados no estado do Maranhão. Foram obtidos um percentual de 82,3%, 65,2% e 25% de amostras positivas na qPCR para M. suis nos municípios de Itapecuru Mirim, Santa Rita e Rosário, respectivamente. Este estudo mostra que M. suis circula entre os suínos de criações extensivas no estado do Maranhão.

Correlação entre o perímetro torácico, peso corporal e a circunferência escrotal de caprinos da raça anglo-nubiano observados na 59ª EXPOEMA / Correlation between scrotal circumference and thorax circumference and between scrotal circumference and body weight of Anglo-Nubian bucks from a livestock exhibition in Maranhão, Brazil

This study aimed to evaluate the correlation between the scrotal circumference and thoraxcircumference and between the scrotal circumference and body weight of Anglo-Nubian bucks from a livestockexhibition in Maranhão, Brazil. 15 bucks were equally divided into three groups according their ages: G1 with9 to 12 months old animals –, G2 with 14 to 19 months old animals –, and G3 with 22 to 29 months oldanimals. The measurements were taken while the event was happening. The data was subjected to thecorrelation coefficient of Pearson. The measures of the scrotal circumference had a positive correlation with thethorax circumference in the groups G1 and G2. G1 group also presented a positive correlation for the bodyweight. However, in this study, it was observed a negative correlation between the scrotal circumference andthorax circumference measures for the animals from the G2 and G3 groups.(AU)

Condição metabólica e integridade das membranas, plasmática e acrossomal, da célula espermática bovina após criopreservação em diferentes meios diluidores acrescidos com diferentes tipos de insulina / Metabolic condition and integrity of membranes, and plasma acrosomal, cell bovine sperm after cryopreservation in different extenders greater with different kinds of insulin

Seminal plasma is a fluid with essential role in sperm functions in vivo, from ejaculation to fertilization.Among the hormones present in this medium, insulin stands out, due to its key role in the structure and motility,favoring fertilization. Insulin acts as preservation factor not able sperm and may be the crux of spermpreservation when using cryogenic processes. This study aimed to evaluate the metabolic and structuralcondition of bovine sperm cells after freezing using extenders plus different types of insulin and different types ofegg yolk. Our results showed that sperm motility was not affected by treatments used (insulin and yolk). Inconclusion, the type of egg yolk, the paw (Anas platyrhynchos), increased the viability of cryopreserved spermcells, regardless of the type of insulin used.(AU)

Efeito da somatotropina recombinante bovina (rbST) no perfil plasmático da progesterona, estradiol, hormônio do crescimento e IGF-1 de fêmeas bovinas / Effect of recombinant bovine somatotropin (rbST) on the plasma levels of progesterone, estradiol, growth hormone and IGF-1 of females bovines

This study aimed to evaluate the effect of recombinant bovine somatotropin on the plasma levels ofprogesterone, estradiol, growth hormone and IGF-1. 8 cows Girolando were divided into three groups: control,250 mg rbST and 500 mg rbST. Blood samples were collected 60 days prior to aspiration, in 12-day intervals,with the samples taken on the same days of rbST applications. For hormone measurements, ELISA commercialkits were used. The hormone concentrations were subjected to the normal pattern Cramer-von Mises and foraverage of comparison, the test Student-Newman-Keuls (SNK) was used, with 5% probability. It was notpossible to measure the plasma concentration of GH hormone. The rbST administration had no effect on plasmaconcentrations of progesterone, estradiol and IGF-1 among the groups treated with 250 mg and 500 mg and theuntreated group, there was also no significant variance between the days of application and OPUs.(AU)