ABSTRACT
Abstract The objective of this study was to standardize and validate the dot-blot test for the serological diagnosis of bovine brucellosis, compare the results with those found in the 2-mercaptoethanol (2-ME) and complement fixation test (CF), and estimate the relative sensitivity and specificity of the dot-blot compared to these tests. Fifty bovine blood serum samples were used for the test standardization, and 1315 samples were used for evaluation and comparison between the tests; the results were compared using the Kappa indicator. At the end of standardization, it was established as optimal for the antigen obtained from Brucella abortus B19 after passing through a microorganism rupture process, the blood serum samples diluted at 1:100, and the conjugate at 1:30,000. The comparison of the dot-blot results with 2-ME showed Kappa index of 0.9939, sensitivity of 99.48%, and specificity 99.91%, with CF, Kappa index of 0.8226, sensitivity 100% and specificity 95.32%. Using the combination of the test results 2-ME and CF to establish the true condition of the animal, the dot-blot showed relative sensitivity of 100%, and relative specificity of 99.91%. The evaluated test proved to be effective and reliable, besides being easy to handle and interpret the results.
Subject(s)
Animals , Cattle , Brucella abortus/isolation & purification , Brucellosis/veterinary , Serologic Tests/methods , Cattle Diseases/diagnosis , Antibodies, Bacterial/blood , Brucella abortus/immunology , Brucellosis/diagnosis , Brucellosis/microbiology , Brucellosis/blood , Serologic Tests/instrumentation , Cattle Diseases/microbiology , Cattle Diseases/blood , Sensitivity and SpecificityABSTRACT
Aims: The activation of cellular and humoral immunity depends upon nature of antigens. Complex proteins like bacterial outer membrane proteins (OMP) usually successfully activate both humoral and cellular immunity. Whereas antigens like bacterial lipopolysaccharides (LPS) usually elicit T-independent immunity i.e. humoral immunity without the activation of cellular immune wing. The present study was under taken to evaluate the comparative immunologic behavior of both the important molecules (bacterial lipopolysaccharide and outer membrane proteins) of Pasteurella multocida alone and in combination in bovine calves in field conditions. Methodology and results: Pasteurella multocida was isolated, purified and identified from an outbreak by mean of culture and biochemical methods. The pathogenicity of the confirmed isolates was done in rabbits (Oryctolagus cuniculus) on the principles of Koch’s postulates. Alum based vaccine against P. multocida was prepared and antibody titer against Outer membrane protein (OMP) and lipopolysaccharides (LPS) were determined by complement fixation test (CFT). The results showed that the antibody titer against OMP and LPS in whole culture vaccine is significantly higher than the respective tested vaccines. These results concluded that OMP no doubt is an active T-dependent immunogenic molecule but its immunogenicity increases many times when combined with LPS in whole culture vaccine. Conclusion, significance and impact of study: Lipopolysaccharides (LPS) in combination with outer membrane proteins (OMP) synergistically boost the humoral immune response in vaccinated animal.
ABSTRACT
FcRn (neonatal Fc receptor) plays an important role in IgG transportation, antigen presentation and signal transmission. In this study, the complement fixation test and flow cytometry test were performed to verify whether the heterologous antibody could be transmitted to the serum or leukocyte with FcγR (Fc gamma receptor) across the intestinal mucosa. The results showed that rabbit anti-bovine IgG could be detected in both the serum and the leukocytes, which indicated that the heterologous antibody could transport across the intestinal mucosa to enter the blood and be effectively delivered to the leukocytes with FcγR. In addition, the results also showed that the rabbit anti-bovine IgG still could be detected in the leukocyte group (P=0.044<0.05) after 21 days. It indicated that the rabbit IgG could exist in the body for a long term (up to 21 days) after being transported to the cells containing FcγR.
ABSTRACT
Objetivou-se com este estudo realizar o primeiro inquérito soro-epidemiológico para o vírus da maedi-visna e Chlamydophila spp. em 12 rebanhos de ovinos do Município de Uberlândia, MG. Foram utilizadas 334 amostras de soro sanguíneo de ovinos e aplicou-se um inquérito epidemiológico a cada propriedade. Os testes realizados para a pesquisa de anticorpos contra o vírus da maedi-visna e Chlamydophila spp. foram imunodifusão em gel de ágar (IDGA) e reação de fixação do complemento (RFC), respectivamente. Não foram detectados ovinos reagentes para maedi-visna. Verificou-se uma prevalência de 3,3% para Chlamydophila spp., com títulos variando de 32 a 64. Não houve diferença estatística significativa (p > 0,05) para os fatores de risco analisados. Ressalta-se a importância dos sistemas de vigilância epidemiológica para atuar no controle dessas infecções, evitando a introdução do vírus da maedi-visna e uma maior propagação da Chlamydophila spp. neste município.
The aim of this study was to carry out the first investigation into the serological prevalence of maedi-visna virus and Chlamydophila spp. on 12 sheep breeding farms in Uberlândia County, MG, Brazil. A total of 334 blood serum samples were used and an epidemiological survey was completed by each farm. The tests to detect maedi-visna and Chlamydophila spp. antibodies were an agar gel immunodiffusion (AGID) and a complement fixation test (CFT), respectively. None of the sheep were reactive to maedi-visna. The detection of antibodies against Chlamydophila spp. was 3.3%, with titers varying from 32 to 64. There was no statistically significant difference (p > 0.05) in regard to the risk factors analyzed. The importance of epidemiological surveillance systems to aid in the control of these infections is emphasized, in order to avoid the introduction of maedi-visna virus and a wider spread of Chlamydophila spp. in this county.
Subject(s)
Animals , Sheep/virology , Chlamydia Infections/epidemiology , Visna-maedi virus/isolation & purification , Pneumonia, Progressive Interstitial, of Sheep/epidemiology , Brazil , Immunodiffusion/veterinary , Abortion, Veterinary/etiologyABSTRACT
Objetivou-se com este estudo identificar os fatores de risco associados à infecção por Chlamydophila abortus em caprinos e ovinos nas regiões do Litoral/ Zonada Mata e Agreste do Estado de Pernambuco. Foram colhidas 290 amostras de soros para pesquisa de anticorpos contra clamídia em 12 propriedades. Para identificar os fatores de risco foram aplicados questionários junto aos proprietários. A freqüência de animais soro-reagentes nos rebanhos estudados foi de 10,3%, sendo 12,0% para caprinos e 8,1% para ovinos, identificando-se 11/12 (91,6%) focos da infecção. Registra-se a primeira ocorrência de anticorpos anti-C. abortus em caprinos no estado de Pernambuco e em ovinos no Brasil. Os fatores derisco associados à infecção em caprinos foram a raça (OR=9,10) e o manejo intensivo (OR=6,41) e para ovinosnão foram encontradas associações significativas para nenhum fator analisado. Concluiu-se que a infecção porChlamydophila sp. encontra-se disseminada em criações de caprinos e ovinos da Zona da Mata e Agreste do Estado de Pernambuco. Medidas de controle devem ser implantadas nos criatórios estudados, enfocando principalmente os fatores de risco identificados neste estudo para reduzir a possibilidade de infecção por este agente.
The study aimed to identify risk factors associated with Chlamydophila abortus infection in sheep and goats from the Litoral/Zona da Mata and Agreste region of Pernambuco state, Brazil. Serum samples (n=290) were analyzed to detect Chlamydophila spp. antibodies in 12 farms. Questionnaires were applied to identify risk factors. Frequency ofserum-reactive animals were 10.3% (12.0% in ewes and 8.1% in goats) and 1/12 (91.6%) infection focuses were identified. This is the first report on anti-Chlamydophhila abortus antibodies in goats and sheep in Pernambuco and Brazil, respectively. Risk factorsassociated with goat infection were breed (OR=9.10) and management (OR=6.41). No significant associations in any of the analyzed factors were found for sheep. In summary, Chlamydophila sp. infection is disseminated in sheep and goat herds in the region. Control measures should be established, focusing primarily risk factor identified in this study, to reduce the possibility of infection by the agent.
Subject(s)
Animals , Chlamydophila/isolation & purification , Goats , Chlamydophila Infections/epidemiology , Risk Factors , SheepABSTRACT
Coccidioidomycosis is a fungal infection that results from inhaling the airborne arthroconidia of the Coccidioides species. It is an endemic disease in the southwest part of North America and rarely diagnosed in Korea. As tourism to endemic areas and the number of immunocompromised patients have been increasing, the incidence of this infection has increased in non-endemic areas. Treatment is usually successful with antifungal agents; however, recurrence is common. It is difficult to decide when to discontinue the antifungal treatment especially in non-endemic areas where doctors are not familiar with the disease. We report a case of recurrent coccidioidomycosis manifesting as osteomyelitis after the treatment of the patient for disseminated coccidioidal infection. The complement fixation test was a useful tool for the assessment of patient response and to evaluate suspected recurrence.
Subject(s)
Humans , Coccidioides , Coccidioidomycosis , Complement Fixation Tests , Endemic Diseases , Immunocompromised Host , Incidence , Inhalation , Korea , North America , Osteomyelitis , RecurrenceABSTRACT
Coccidioidomycosis is a fungal infection that results from inhaling the airborne arthroconidia of the Coccidioides species. It is an endemic disease in the southwest part of North America and rarely diagnosed in Korea. As tourism to endemic areas and the number of immunocompromised patients have been increasing, the incidence of this infection has increased in non-endemic areas. Treatment is usually successful with antifungal agents; however, recurrence is common. It is difficult to decide when to discontinue the antifungal treatment especially in non-endemic areas where doctors are not familiar with the disease. We report a case of recurrent coccidioidomycosis manifesting as osteomyelitis after the treatment of the patient for disseminated coccidioidal infection. The complement fixation test was a useful tool for the assessment of patient response and to evaluate suspected recurrence.
Subject(s)
Humans , Coccidioides , Coccidioidomycosis , Complement Fixation Tests , Endemic Diseases , Immunocompromised Host , Incidence , Inhalation , Korea , North America , Osteomyelitis , RecurrenceABSTRACT
La brucelosis es considerada como una de las enfermedades infecciosas más importantes enColombia y el mundo por sus implicaciones en la salud pública, y sus repercusiones productivas en el ámbito pecuario, dejando grandes pérdidas a la economía nacional. Considerando que existe poca información sobre su prevalencia en el ámbito regional, en el transcurso de los años 2001-2002 se realizó un estudio sobre la prevalencia de la brucelosis bovina, equina y humana (personal con alto riesgo de infección) en el departamento de Caldas. Para el desarrollo del trabajo se utilizaron como pruebas diagnósticas, las técnicas rosa de Bengala y ELISA Indirecta; además se realizaron pruebas confirmatorias como ELISA competitiva y fijación de complemento para los equinos. Los resultados obtenidos en 2.434 muestras de bovinos mostraron una baja incidencia para brucelosis (0,6%), que correspondió a 15 animales hembras, distribuida en las cuatro zonas del departamento de la siguiente manera: oriente 0,24% (6 bovinos: 3 de La Dorada y 3 de la Victoria); centro-sur 0,16% (4 bovinos: 2 de Manizales, 1 de Belacázar y 1 de Villamaría); norte 0,12% (3 bovinos: 1 de Neira, 1 de Aranzazu y 1 de Salamina) y occidente 0,08% (2 bovinos, ambos en Risaralda), encontrándose animales positivos en rangos de edad de 18 a 90 meses. No se hallaron equinos reactores positivos utilizando las pruebas confirmatorias. Los resultados obtenidos en 676 muestras de humanos, arrojaron una baja prevalencia (0,14%), correspondiente a un matarife, con síntomas compatibles para brucelosis. La técnica rosa de Bengala aumentó la sensibilidad al agregar el doble de la cantidad de suero con respecto al antígeno Rosa de Bengala.
Brucellosis is considered one of the most important infectious diseases in Colombia and the world due to its implications in public health, and its productive consequences in the livestock field that is producing big losses in the economy of the country. Since there is limited information on Brucellosis in Caldas, in 2001-2002 a prevalence study on bovine, equine and human (high risk personnel) Brucellosis was carried out in the department of Caldas. For the development of the research the following diagnostic tests were used: Bengal Rose Test, indirect ELISA, as well as confirmatory tests, such as competitive ELISA and Complement Fixation, for the horses. The results obtained in 2,434 bovine samples shown a low incidence of Brucellosis (0.6%), corresponding to 15 females, distributed in the four zones of the department as follows: east zone with 0.24% (3 in La Dorada and 3 in La Victoria); central-south zone with 0.16% (2 in Manizales, 1 in Belalcazar and 1 in Villamaría); north zone with 0.12% (1 in Neira, 1 in Aranzazu and 1 in Salamina); and west zone with 0.08% (2 in Risaralda). The animals that tested positive were between 18 to 90 months of age. No equine reactors tested positive with the confirmatory test. The results obtained in 676 human samples showed a low prevalence (0.14%), corresponding to a slaugtherman with symptoms associated with Brucellosis. The Bengal Rose Test increased the sensibility when adding twice as much serum, regarding the Bengal Rose antigen.
Subject(s)
Cattle , Brucellosis , Cattle , Horses , Humans , PrevalenceABSTRACT
Para el diagnóstico de la brucelosis bovina en muestras de sangre y/o leche, se comparó la reacción en cadena de la polimerasa (PCR) con el aislamiento in vitro de Brucella abortus, las pruebas serológicas defijación del complemento (FC) e inmunoenzimáticas de competición (ELISA-C) en suero e indirecto (ELISA-I) en leche. Se analizaron muestras de vacas lecheras de un rebaño infectado “A”, vacunadas con B. abortus cepa 19 antes de los 8 meses de edad y revacunadas con B. abortus cepa RB51 como adultas (n= 99) y de otro “B”, libre de brucelosis (n=100), como control. En A, la PCR identificó 14 vacas infectadas con B. abortus: nueve con cepa silvestre y cinco con cepa silvestre y RB51. No se identificó B. abortus cepa 19. El biotipo 1 se aisló en un caso. Las 14 vacas infectadas con la cepa silvestre resultaron positivas en las tres pruebas serológicas. En B, por PCR no se identificó Brucella. Las pruebas serológicas mostraron una sensibilidad del 100% respecto de PCR. La especificidad para FC, ELISA-C y ELISA-I fue del 100%, 99% y 95%, respectivamente. Se concluye que la PCR sería útil como complemento de las pruebas serológicas o cuando no hay un resultado concluyente.
The diagnosis of bovine brucellosis using PCR in blood and milk samples from two dairy herds were compared to in vitro isolation, complement fixation test (CF), competitive ELISA (C-ELISA) in serum, and indirect ELISA (I-ELISA) in milk. Samples were obtained from 99 cows vaccinated with Brucella abortus strain 19, from a naturally infected herd (A), whose cows were also vaccinated with B. abortus strain RB51 as adults, and 100 from brucellosis free herd (B). In herd A, PCR identified 14 B. abortus infected cows: nine infected with wild type, and five with wild type and RB51, B. abortus S 19 was not identified. B. abortus biotype 1 was isolated from one cow. All cows infected with a wild strain of B. abortus were positive in serologic tests. Brucella was not found in herd B using PCR. Serological test showed 100% sensitivity related to PCR. The specificity for CF, C-ELISA and I-ELISA was 100%, 99% and 95% respectively. PCR could be useful to identify Brucella biotypes and to complement serologic tests.