The aim of the current study was to verify if cattle vaccinated against leptospirosis may react in diagnostic tests for brucellosis. Sixty cows were divided into 5 groups, each comprising 12 animals. Four groups were given different vaccines against leptospirosis, while the control group received only saline. Two doses of vaccine were given, as recommended by the manufacturers. Serum samples were collected on the first day of immunization (day 0) and on postvaccination days 7, 14, 21, 28, 35, 42, 49, 56, 96, and 126. All the serum samples were tested for brucellosis and leptospirosis. Twenty animals were reactive at least once to the Rose Bengal test, but by day 96, no further reactions were elicited by this test. Twenty-six samples were reactive to the Rose Bengal test, but only 7 remained positive in confirmatory tests: 1 to the 2-mercaptoethanol test, 2 to the fluorescence polarization assay, and 6 to indirect enzyme-linked immunosorbent assays. None of the samples was reactive in the complement fixation test. None of the animals in the control group was reactive. A significant difference was found between the control group and the groups vaccinated against leptospirosis, according to Fisher exact test. However, the groups were found to respond independently of the vaccine brand. The results indicate that cattle vaccinated against leptospirosis may show reactivity on screening tests for brucellosis.
ABSTRACT:Leptospirosis is a zoonotic disease that occurs worldwide, affecting pets and livestock that can be a source of infection for humans, considered therefore an important disease in public health. Thus, an investigation was made into the occurrence of anti-Leptospira agglutinins in several animal species on a farm located in the municipality of Uberlândia, state of Minas Gerais, Brazil. A total of 172 blood serum samples were collected from five species: cattle, dogs, goats, horses and sheep. The diagnosis was performed by means of the microscopic agglutination test (MAT), using a collection of 12 live antigens. Of the 172 animals evaluated, 62 (36%) were seroreactive, and the most frequent serovars in each species were: Hardjo (54.54%), Icterohaemorrhagiae (27.27%), and Autumnalis (18.18%) in cattle; Icterohaemorrhagiae (37.5%), Autumnalis (31%), Canicola (18.7%) and Hardjo (12.5%) in dogs; Grippotyphosa (75%) and Hardjo (25%) in goats; Icterohaemorrhagiae (50%), Grippotyphosa (37.5%) and Hardjo (12.5%) in horses, and Hardjo (34.78%); Grippotyphosa (26.08%); Bratislava (17.39%), Autumnalis (17.39%) and Icterohaemorrhagiae (4.4%) in sheep. It is believed that the occurrence of anti-Leptospira agglutinins can be traced back to a source of infection that is common to these species, or that the agent was probably transmitted between them because they live in close proximity to each other.
RESUMO: Objetivou-se com este estudo comparar diferentes métodos complementares de diagnóstico (macroscópico, histopatológico, sorológico e molecular) da tuberculose, em bovinos naturalmente infectados. O trabalho deu-se por meio de amostras colhidas em abate sanitário de 40 bovinos reagentes no teste cervical comparativo (TCC) para tuberculose. A inspeção macroscópica post mortem das carcaças foi acompanhada de colheita de amostras de muco nasal, sangue e tecido (fígado, pulmão e linfonodo mediastínico) para realização do exame de reação em cadeia pela polimerase (PCR), de Enzyme-Linked Immunosorbent Assay (ELISA) e histopatológico com colorações de hematoxilina-eosina (HE) e Ziehl-Neelsen (ZN), respectivamente. Dos 40 bovinos inspecionados no abate, 22 (55%) animais apresentaram algum tipo de lesão macroscópica sugestiva de tuberculose. Nos achados histopatológicos visualizados em HE, 13 (32,5%) carcaças apontaram alterações histológicas, sendo 6 (15%) nos linfonodos mediastínicos, 5 (12,5%) no fígado e 3 (7,5%) no pulmão. Não foi observada a presença de bacilos álcool-ácido resistentes em nenhuma das amostras avaliadas. O ensaio sorológico de ELISA/IDEXX(r) identificou um (2,5%) animal reagente, e o teste de PCR detectou DNA de Mycobacterium bovis em uma (2,5%) amostra. Concluiu-se que entre os exames complementares de diagnóstico avaliados nenhum foi capaz de detectar todos os animais que estavam positivos na tuberculinização, porém a associação de diferentes métodos pode garantir a confiabilidade do diagnóstico.
RESUMO:O objetivo deste trabalho foi analisar a frequência de lesões macroscópicas sugestivas de tuberculose em carcaças de bovinos reagentes ao teste intradérmico de tuberculose. Os animais do estudo foram provenientes de rebanhos de exploração leiteira, localizados nos municípios de Perdizes, Patos de Minas, Lagoa Formosa e Uberlândia, Minas Gerais. Foram avaliadas 140 carcaças de bovinos positivos no teste cervical comparativo. Durante o abate sanitário foi realizada inspeção post mortem das carcaças, e as lesões macroscópicas sugestivas de tuberculose foram registradas. Foram avaliados os linfonodos mediastínicos, fígado, pulmão e carcaça. Dos 140 bovinos examinados, 78 (55%) apresentaram algum tipo de lesão macroscópica sugestiva de tuberculose, 38 (49%) ocorreram exclusivamente nos linfonodos mediastínicos, 22 (28%) no fígado e 11 (14%) no pulmão; 5 (6%) carcaças apresentaram lesões no fígado, pulmão e linfonodo mediastínico, e 2 (4%) tiveram lesões no pulmão e linfonodo. Concluem-se que as lesões não visualizadas em bovinos reagentes ao teste tuberculínico podem ocorrer e estariam relacionadas principalmente ao estágio de evolução da doença, tempo insuficiente na inspeção post mortem para detecção, bem como reações inespecíficas para outras micobactérias. PALAVRAS-CHAVE: ABSTRACT:The aim of this study was to analyze the frequency of macroscopic lesions suggestive of tuberculosis in cattle reacting to the tuberculin test. Cattle of this study were from dairy herds located in the cities of Perdizes, Patos de Minas, Lagoa Formosa and Uberlândia, Minas Gerais, Brazil. We evaluated 140 cattle carcasses positive to the cervical comparative intradermal tuberculin test. During the slaughter a post-mortem inspection of carcasses was done, and the lesions suggestive of tuberculosis were recorded. We evaluated the mediastinal lymph nodes, liver, lung and carcass. Of the 140 cattle examined, 78 (55%) carcasses had some kind of macroscopic lesions suggestive of tuberculosis, 38 (49%) occurred only in the mediastinal lymph nodes, 22 (28%) in the liver and 11 (14%) in the lungs; 5 (6%) carcasses showed lesions in liver, lungs and lymph node, and 2 (4%) showed lesions in lung and lymph nodes. We concluded that lesions that were not visualized in bovine tuberculin skin test reagents may occur and are related mainly to the stage of the disease, insufficient time to detect the lesions, and nonspecific reactions to other mycobacteria.
Background: The immunoglobulin Y is a principal antibody current in the blood of hens, which are transferred from the maternal blood serum to the egg yolk. The extraction of IgY from the egg yolk apply animal welfare when compared to the extraction of IgG, reducing the number of animals and prevent a bleeding of hens through the extraction of the IgY from eggs, besides that IgY presenting high specificity for antigenic binding. The objective of this study was to produce specific polyclonal IgY antibodies anti-Brucella abortus by immunizing hens with B19 vaccine and evaluate their reactivity through Buffered Acidified Plate Antigen (BAPA), 2-Mercaptoethanol (2-ME) and indirect ELISA diagnostic tests.Materials, Methods & Results: Four 25-week-old White Leghorn hens were immunized, two of them comprising the control group (Group 1) with phosphate-buffered saline (PBS) with adjuvant, and the others two immunized with B19 vaccine (Brucella abortus vaccine strain B19), representing the Group 2. The immunizations occurred six times with a 15-day interval between each. Blood samples were taken biweekly (seven times); and daily, the eggs were collected for 13 weeks, the first collection of blood and eggs, performed one week before the first immunization of each group. The IgY was purified from egg yolk, using the method of dilution in acid water and precipitation with ammonium sulfate for delipidation. BAPA, 2-ME and ELISA tests performed to verify the specificity of IgY confirmed the reactivity of polyclonal antibodies specific to the antigen used both in blood serum samples and in the purified egg yolks. The hens from the control group did not present reactivity in the diagnostic tests used, which was already expected, since no antigen was used in any of their immunizations. Hens immunized with the Brucella abortus B19 vaccine produced detectable reactive antibodies in the three tests used on blood serum and IgY samples extracted from the egg yolk. In Group 2 (vaccine B19), blood serum samples started to react one week after the first inoculation, and the IgY samples extracted from the egg yolk were reagent two weeks after serum IgY appear reactivity, showing the transfer of specific antibodies to the egg yolk, was late.Discussion: Although the transfer of serum Igy to egg yolk was late when compared to others authors which found that the transovarian passage of immunoglobulin Y occurred in approximately three to six days after IgY being detected in blood serum, the results of this study showed the occurrence of the transfer of blood serum IgY anti-Brucella abortus to egg yolk of hens immunized with B19 vaccine, the same found by others researches found the same results with others antigens. Thus, it can be concluded that immunoglobulins Y produced in this study can be used as specific antibodies in diagnostic tests for the detection of the Brucella abortus antigen, in addition, this process guarantees the welfare of the animal, since it avoids bleeding and it is possible to obtain high concentrations of antibodies directly from the hen egg, which is a great advantage, because IgY can be easily isolated from the egg yolk by the precipitation technique discarding the need of invasive and painful procedures that involve bloody interventions to obtain the serum antibodies like occur in mammals for extraction of IgG.
Objetivou-se verificar se galinhas imunizadas com uma solução de Leptospira interrogans inativadas e proteínas de membrana externa do sorovar Hardjo, poderiam produzir anticorpos policlonais específicos anti-leptospiras, detectáveis em testes ELISA. Foram imunizados oito galinhas com 25 semanas de idade, da raça White Leghorn, sendo três imunizadas com uma suspensão de leptospiras inativadas, três com uma solução de proteínas de membrana externa extraída do sorovar Hardjo e duas controle. Coletas de sangue foram realizadas quinzenalmente e de ovos diariamente. A IgY foi purificada a partir da gema dos ovos utilizando para a delipidação o método de diluição em água ácida e a precipitação com sulfato de amônio. Nos testes ELISA realizados para verificar a especificidade da IgY, foi demonstrada a produção de anticorpos anti-Leptospira, tanto no soro quanto nas gemas purificadas. O pico de produção de anticorpos específicos ocorreu na 5º semana após a primeira imunização. Ficou demonstrada a possibilidade da indução da produção de anticorpos específicos em galinhas imunizadas com leptospiras do sorovar Hardjo inativadas, bem como, com proteínas de membrana externa (PME) extraidas desse sorovar. As galinhas imunizadas com uma suspensão de leptospiras inativadas ou com PME de Leptospira interrogans do sorovar Hardjo produziram anticorpos reativos a PME Hardjo detectáves por teste ELISA.
RESUMOA Tuberculose (TB) e a Leucose Enzoótica dos Bovinos (LEB) são doenças infectocontagiosas, caracterizadas pela evolução crônica e pelos prejuízos gerados à pecuária bovina. Essas doenças comprometem o desempenho produtivo dos rebanhos, causando condenações de carcaças em frigoríficos e restringindo o comércio de animais, além do aumento dos custos com serviços veterinários. A TB, além da importância em saúde pública, causa reduções de até 25% na produtividade animal. O vírus da LEB está associado ao desencadeamento de bacterioses oportunistas. Admite-se, que o comprometimento da integridade do sistema imunitário orgânico pela ação imunodepressora do vírus, que penetra e incorporase no genoma linfócitário por tempo indeterminado aumenta a susceptibilidade do hospedeiro a outras infecções. Objetivou-se com este trabalho, avaliar a ocorrência da Tuberculose Bovina e Leucose Enzoótica dos Bovinos (LEB), em um rebanho bovino leiteiro. Foram examinados 316 bovinos de ambos os sexos, com idade entre 6 meses e 16 anos, pelo teste alérgico-cutâneo, o exame utilizado foi o Teste Cervical Comparativo (TCC). Para o diagnóstico da LEB, foram avaliados 85 animais, escolhidos aleatoriamente, empregou-se a técnica de Imunodifusão em Gel de Ágar (IDGA) segundo o protocolo do fabricante do antígeno TECPAR, por meio de um substrato de difusão gelatinoso, utilizando o antígeno glicoprotéico (gp 51), extraído do envelope do Vírus da Leucose Bovina. Foram reagentes no TCC 3,16% (10/316), e 5,06% (16/316) considerados inconclusivos. Já no IDGA foram diagnosticados 56,32% (48/85) bovinos positivos. Conclui-se que ambas as doenças estão presentes no rebanho, e que a alta porcentagem de positivos para Leucose, podem tornar os animais susceptíveis à Tuberculose, uma vez que a LEB leva o animal a um quadro de imunosupressão.
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