RESUMO
Background: The aim of the present study was to describe the presence of co-infection by Toxoplasma gondii and Neospora caninum in goats reared in extensive systems from Mexico. Materials and Methods: A cross-sectional study was conducted to determine the frequency of T. gondii and N. caninum, by detecting antibodies to each parasite by mean commercial ELISA kits. A total of 176 blood samples were randomly collected from mature females reared in extensive system herds from 20 municipalities of state of Guanajuato, Mexico. Results: The general seroprevalence was 23.9 and 21.0% for T. gondii and N. caninum, respectively, while co-infection rate was 3.6%. For geographic and environmental variables, no differences were observed among T. gondii and coinfection; however, it was observed that altitude, annual precipitation, annual average temperature, and rainy period showed significant differences with N. caninum seropositive goats. Conclusion: The seroprevalence of both parasites was appreciated in most of the studied herds. The present study is the first report of T. gondii and N. caninum co-infection in goats from extensive herds in Mexico.
Assuntos
Coccidiose , Coinfecção , Doenças das Cabras , Cabras , Neospora , Toxoplasma , Toxoplasmose Animal , Animais , Coccidiose/veterinária , Coccidiose/epidemiologia , Coccidiose/parasitologia , México/epidemiologia , Toxoplasma/imunologia , Doenças das Cabras/epidemiologia , Doenças das Cabras/parasitologia , Toxoplasmose Animal/epidemiologia , Toxoplasmose Animal/parasitologia , Coinfecção/epidemiologia , Coinfecção/veterinária , Coinfecção/parasitologia , Estudos Soroepidemiológicos , Feminino , Estudos Transversais , Anticorpos Antiprotozoários/sangueRESUMO
Despite global efforts to assess the early response and persistence of SARS-CoV-2 antibodies in patients infected with or recovered from COVID-19, our understanding of the factors affecting its dynamics remains limited. This work aimed to evaluate the early and convalescent immunity of outpatients infected with SARS-CoV-2 and to determine the factors that affect the dynamics and persistence of the IgM and IgG antibody response. Seropositivity of volunteers from Mexico City and the State of Mexico, Mexico, was evaluated by ELISA using the recombinant receptor-binding domain (RBD) of the SARS-CoV-2 Spike protein for 90 days, at different time points (1, 15, 45, 60, and 90 days) after molecular diagnosis (RT-qPCR). Gender, age range, body mass index (BMI), comorbidities, and clinical spectrum of disease were analyzed to determine associations with the dynamics of anti-SARS-CoV-2 antibodies. On 90 days post-infection, individuals with moderate and asymptomatic disease presented the lowest levels of IgM, while for IgG, at the same time, the highest levels occurred with mild and moderate disease. The IgM and IgG levels were related to the clinical spectrum of disease, BMI, and the presence/absence of comorbidities through regression trees. The results suggest that the dynamics of anti-SARS-CoV-2 IgM and IgG antibodies in outpatients could be influenced by the clinical spectrum of the disease. In addition, the persistence of antibodies against SARS-CoV-2 could be related to the clinical spectrum of the disease, BMI, and the presence/absence of comorbidities.
Assuntos
COVID-19 , Humanos , SARS-CoV-2 , Anticorpos Antivirais , Imunoglobulina G , Imunoglobulina M , ImunidadeRESUMO
Brucella canis, a Gram-negative coccobacilli belonging to the genus Brucellae, is a pathogenic bacterium that can produce infections in dogs and humans. Multiple studies have been carried out to develop diagnostic techniques to detect all zoonotic Brucellae. Diagnosis of Brucella canis infection is challenging due to the lack of highly specific and sensitive diagnostic assays. This work was divided in two phases: in the first one, were identified antigenic proteins in B. canis that could potentially be used for serological diagnosis of brucellosis. Human sera positive for canine brucellosis infection was used to recognize immunoreactive proteins that were then identified by performing 2D-GEL and immunoblot assays. These spots were analyzed using MALDI TOF MS and predicted proteins were identified. Of the 35 protein spots analyzed, 14 proteins were identified and subsequently characterized using bioinformatics, two of this were selected for the next phase. In the second phase, we developed and validated an indirect enzyme-linked immunosorbent assays using those recombinant proteins: inosine 5' phosphate dehydrogenase, pyruvate dehydrogenase E1 subunit beta (PdhB) and elongation factor Tu (Tuf). These genes were PCR-amplified from genomic DNA of B. canis strain Oliveri, cloned, and expressed in Escherichia coli. Recombinant proteins were purified by metal affinity chromatography, and used as antigens in indirect ELISA. Serum samples from healthy and B. canis-infected humans and dogs were used to evaluate the performance of indirect ELISAs. Our results suggest that PdhB and Tuf proteins could be used as antigens for serologic detection of B. canis infection in humans, but not in dogs. The use of recombinant antigens in iELISA assays to detect B. canis-specific antibodies in human serum could be a valuable tool to improve diagnosis of human brucellosis caused by B. canis.
RESUMO
OBJECTIVES: Hyperinfection or disseminated strongyloidiasis has been frequently reported after transplants and is related to high mortality. This study aimed to screen for strongyloidiasis using serological diagnoses in transplant candidates. METHODS: An ELISA test was performed with filariform larvae of Strongyloides venezuelensis as a source of antigen. RESULTS: In the serum from transplant candidates, anti-Strongyloides IgG antibodies were detected in 35/150 (23.3%) samples by soluble fractions in phosphate buffered saline (PBS), 31/150 (20.7%) samples by soluble fractions in Tris-HCl, 27/150 (18.0%) samples by membrane fractions in PBS and 22/150 (14.7%) samples by membrane fractions in Tris-HCl. CONCLUSIONS: The present results suggest the ELISA test, ideally using soluble fractions of filariform larvae S. venezuelensis in PBS, as an additional strategy for the diagnosis of strongyloidiasis in transplant candidates.
Assuntos
Humanos , Animais , Masculino , Feminino , Criança , Adolescente , Adulto , Pessoa de Meia-Idade , Adulto Jovem , Estrongiloidíase/diagnóstico , Imunoglobulina G/sangue , Transplante de Órgãos , Strongyloides stercoralis/imunologia , Antígenos de Helmintos/imunologia , Estrongiloidíase/parasitologia , Ensaio de Imunoadsorção Enzimática , Anticorpos Anti-Helmínticos/sangue , Biomarcadores/sangue , Programas de Rastreamento , Sensibilidade e Especificidade , Hospedeiro Imunocomprometido , Antígenos de Helmintos/isolamento & purificaçãoRESUMO
Introdución: el Sindrome de Inmunodeficiencia Adquirida (VIH) sigue siendo un importante problema de salud pública a nivel mundial, ya que ha causado más de 34 millones de muertes hasta la fecha. Objetivo: conocer las barreras y facilitadores de un grupo personas que solicitan el test Elisa para el diagnóstico del Virus de Inmunodeficiencia Adquirida (VIH) en la atención primaria de salud, desde la perspectiva de los profesionales de la salud que trabajan en estos lugares. Métodos: estudio cualitativo descriptivo basado en el enfoque de análisis de contenido descrito por Krippendorff. Se realizaron entrevistas en profundidad a 30 profesionales de la salud que trabajan en dos Centros de Salud Familiar (CESFAM) de la comuna de La Pintana, Santiago de Chile. Para mantener el rigor metodológico se utilizaron los criterios de Guba & Lincoln. Resultados: se identificaron factores asociados al acceso oportuno al test de Elisa percibidas por los profesionales, entre ellos algunos que dificultan y otros que facilitan (Estigma social del VIH, burocracia en el proceso de toma del test de Elisa y falta de privacidad en la solicitud del test, relación de confianza entre usuario y profesional, y centralización del examen en los CESFAM). Conclusiones: el conocimiento de los factores tanto que facilitan como que dificultan mencionado por los profesionales que trabajan en centros donde se realiza la toma del test de Elisa debe ser considerada en el diseño e implementación de nuevas estrategias y en la modificación de las existentes, con la finalidad de aumentar el número de personas que accedan al test y con esto mejorar la calidad del cuidado y la satisfacción usuaria(AU)
Introduction: The Acquired Immunodeficiency Syndrome (HIV) continues to be a critic health concern for the worldwide public health, since it has caused up to date more than 34 million deaths. Objective: Know the obstacles and facilitators of a group of people who request the ELISA test for diagnosing the acquired immunodeficiency virus (VIH) in primary health care, from the perspective of the health professional who works in this sites. Methods: Qualitative descriptive study based on the content analysis approach described by Krippendorff. In-deep interviews were conducted on 30 health professionals who work in two Family Health Centers (CESFAM) of La Pintana Town, Santiago de Chile. The Guba & Lincoln criteria were used, in order to maintain the methodologic rigor. Results: We identified factors associated with the timely access to ELISA tests as perceived by the professionals, among then some that make difficult and others that facilitate (HIV social stigma, bureaucracy in taking the ELISA test and the lack of privacy in requesting the test, confidence relationship between the user and the professional, and centralization of the test in the CESFAM's). Conclusions: Knowing both the facilitating and hindering factors mentioned by the professionals who work in centers where the ELISA tests are made should be considered in the design and implementation of new strategies and in the modification of the existing ones, with the aim to increase the number of people who can access the tests and thus to improve the quality of care and the users' satisfaction(AU)
Assuntos
Humanos , Masculino , Feminino , Atenção Primária à Saúde , Síndrome da Imunodeficiência Adquirida/diagnóstico , Síndrome da Imunodeficiência Adquirida/epidemiologia , Barreiras de Comunicação , Promoção da Saúde/métodos , Ensaio de Imunoadsorção Enzimática/métodosRESUMO
Introdución: el Sindrome de Inmunodeficiencia Adquirida (VIH) sigue siendo un importante problema de salud pública a nivel mundial, ya que ha causado más de 34 millones de muertes hasta la fecha. Objetivo: conocer las barreras y facilitadores de un grupo personas que solicitan el test Elisa para el diagnóstico del Virus de Inmunodeficiencia Adquirida (VIH) en la atención primaria de salud, desde la perspectiva de los profesionales de la salud que trabajan en estos lugares. Métodos: estudio cualitativo descriptivo basado en el enfoque de análisis de contenido descrito por Krippendorff. Se realizaron entrevistas en profundidad a 30 profesionales de la salud que trabajan en dos Centros de Salud Familiar (CESFAM) de la comuna de La Pintana, Santiago de Chile. Para mantener el rigor metodológico se utilizaron los criterios de Guba & Lincoln. Resultados: se identificaron factores asociados al acceso oportuno al test de Elisa percibidas por los profesionales, entre ellos algunos que dificultan y otros que facilitan (Estigma social del VIH, burocracia en el proceso de toma del test de Elisa y falta de privacidad en la solicitud del test, relación de confianza entre usuario y profesional, y centralización del examen en los CESFAM). Conclusiones: el conocimiento de los factores tanto que facilitan como que dificultan mencionado por los profesionales que trabajan en centros donde se realiza la toma del test de Elisa debe ser considerada en el diseño e implementación de nuevas estrategias y en la modificación de las existentes, con la finalidad de aumentar el número de personas que accedan al test y con esto mejorar la calidad del cuidado y la satisfacción usuaria(AU)
Introduction: The Acquired Immunodeficiency Syndrome (HIV) continues to be a critic health concern for the worldwide public health, since it has caused up to date more than 34 million deaths. Objective: Know the obstacles and facilitators of a group of people who request the ELISA test for diagnosing the acquired immunodeficiency virus (VIH) in primary health care, from the perspective of the health professional who works in this sites. Methods: Qualitative descriptive study based on the content analysis approach described by Krippendorff. In-deep interviews were conducted on 30 health professionals who work in two Family Health Centers (CESFAM) of La Pintana Town, Santiago de Chile. The Guba & Lincoln criteria were used, in order to maintain the methodologic rigor. Results: We identified factors associated with the timely access to ELISA tests as perceived by the professionals, among then some that make difficult and others that facilitate (HIV social stigma, bureaucracy in taking the ELISA test and the lack of privacy in requesting the test, confidence relationship between the user and the professional, and centralization of the test in the CESFAM's). Conclusions: Knowing both the facilitating and hindering factors mentioned by the professionals who work in centers where the ELISA tests are made should be considered in the design and implementation of new strategies and in the modification of the existing ones, with the aim to increase the number of people who can access the tests and thus to improve the quality of care and the users' satisfaction(AU)
Assuntos
Humanos , Masculino , Feminino , Atenção Primária à Saúde , Síndrome da Imunodeficiência Adquirida/diagnóstico , Síndrome da Imunodeficiência Adquirida/epidemiologia , Barreiras de Comunicação , Promoção da Saúde/métodos , Ensaio de Imunoadsorção Enzimática/métodosRESUMO
BACKGROUND: The ELISA format for measuring carcinoembryonic antigen (CEA) serves as a reference standard against which other assays are compared. Because the World Health Organization (WHO) increasingly recommends the use of serum CEA as a diagnostic tool for cancer, it is relevant to explore the reliability of the new decentralized CEA point-of-care-testing (POCT) technologies that are available to physicians and patients, in compliance with mandates of the clinical laboratories' regulatory agencies. METHODS: Electrochemical immunoassay (ECIA) based on trace lead (Pb) analysis by anodic stripping techniques using sandwich-type immunocomplex conjugates: (MB)Ab/AgCEA/Ab(PbS), and a commercial ELISA test system with optical transmission. RESULTS: The ECIA provides better analytical performance than does the ELISA. The within assay precision coefficient of variance (%CVw) of the ECIA is lower than the value recommended by the Hong Kong Association of Medical Laboratories (HKAML), and the recoveries of CEA at 1.0, 5.0, 10.0, 25.0 and 50.0 ng/ml are in the range of 99-110% for control serum samples. The ECIA showed a minimal positive bias of 0.0267 ± 0.3270 ng/ml (P=0.9389). CONCLUSIONS: The proposed CEA screening technology can be practically employed for decentralized clinical analysis of CEA in human serum. Therefore, it can be viewed as a control method for personalized therapy.
Assuntos
Biomarcadores Tumorais/sangue , Antígeno Carcinoembrionário/sangue , Técnicas Eletroquímicas , Ensaio de Imunoadsorção Enzimática , Bioestatística , Detecção Precoce de Câncer , Técnicas Eletroquímicas/instrumentação , Ensaio de Imunoadsorção Enzimática/instrumentação , Humanos , Tamanho da Partícula , Propriedades de SuperfícieRESUMO
OBJECTIVES: Infection caused by Helicobacter pylori (H. pylori) is one of the most common causes of chronic infection in the world. The presence of the infection is strongly associated with the neoplasia of the gastrointestinal tract, and its diagnosis is easily made by means of invasive or non-invasive methods. Among such methods, the H. pylori antigen detection in stool through ELISA technique is easily performed and it is an alternative to endoscopy in children, since this exam is not usually indicated in this age group. The aim of the current study is to establish the standardization of the ELISA method for the detection of H. pylori in stool specimens in Brazil. DESIGN AND METHODS: Patients between 18 and 70 years of age were randomly selected in the gastroenterology ambulatory center at Faculdade de Medicina do ABC between 2007 and 2009. They all answered a questionnaire to investigate possible dyspeptic symptoms and then underwent endoscopy and detection of H. pylori through no more than 4 methods. Besides the gastric biopsy, established as the gold standard test, the urease test, the stool ELISA test and serology were also methods applied. RESULTS: The sensitivity and specificity of the exams in this sample were respectively 87.2% and 44% for the stool ELISA test, 41.9% and 64% for serology, 65.6% and 58.8% for the urease test and 100% and 80.8% for the clinical analysis. CONCLUSIONS: The ROC curve showed a good correlation between the compared methods. In Brazil the standardization of the ELISA test for the detection of H. pylori in stool specimens constitutes a non-invasive diagnostic alternative.
Assuntos
Ensaio de Imunoadsorção Enzimática/normas , Fezes/microbiologia , Infecções por Helicobacter/diagnóstico , Helicobacter pylori/isolamento & purificação , Adolescente , Adulto , Antígenos de Bactérias/sangue , Proteínas de Bactérias/análise , Biópsia , Brasil , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Infecções por Helicobacter/microbiologia , Helicobacter pylori/imunologia , Humanos , Masculino , Pessoa de Meia-Idade , Sensibilidade e Especificidade , Urease/análiseRESUMO
Enzootic bovine leukosis (EBL) is a retroviral infection that causes persistent lymphocytosis and lymphosarcoma in cattle. The economic importance of infection by bovine leukemia virus (BLV) is due to several factors, including losses in exportation, treatment of secondary infection, and reduction in dairy production. To facilitate the development of a national test that is sensitive, simple, and applicable on a large scale, this work aimed to produce and characterize monoclonal antibodies (mAbs) against gp51 protein from BLV for use in an enzyme-linked immunosorbent assay (ELISA) test. Two hundred seventy-four hybridomas were generated, from which 37 were mAbs secretory clones screened by indirect ELISA. The specificity of the mAbs generated against gp51 was verified by Western blot analysis, and the isotypes were characterized for isotyping in IgG1 and IgM. To evaluate the test, 250 sera were tested by agar gel immunodiffusion and mAb-ELISA. The values obtained for the mAb-ELISA test were 95% sensitivity and 90% specificity.
RESUMO
Objetivo. Determinar los agentes infecciosos asociados causalmente a la presentación de Diarrea Neonatal Bovina (DNB) en terneros menores de 5 semanas de vida, procedentes de fincas lecheras y de producción mixta de la Sabana de Bogotá. Materiales y métodos. Se seleccionaron por conveniencia 21 fincas, se realizó seguimiento de 620 terneros desde el nacimiento hasta las 5 semanas de edad, se tomaron muestras de materia fecal de los animales que presentaron cuadro clínico de diarrea y de terneros clínicamente sanos como controles pareados. Se realizaron pruebas de ELISA para diagnóstico de E.coli F5, Rotavirus, Coronavirus, Cryptosporidium sp., y Salmonella sp., prueba de Ritchie para diagnóstico de Giardia sp., y tinción de Ziehl Neelsen modificada para Cryptosporidium sp. Se evaluó la asociación epidemiológica entre los agentes y la presentación de diarrea usando prueba de c2, seguido de un modelo de regresión logística (p<0.05). Resultados. Se encontró en la prueba de ELISA que de la totalidad de las muestras, 51 (38.3%), 26 (19.7%), 10 (7.5%) y 1 (0.75%) fueron positivas a Cryptosporidium sp., rotavirus, E coli F5 y coronavirus, respectivamente. Los animales positivos a Rotavirus por la prueba de ELISA y a Cryptosporidium sp., por la técnica de Ziehl Neelsen modificada tuvieron 2.6 y 7.0 veces mayor probabilidad que los demás animales de presentar DNB, respectivamente. Conclusiones. Los resultados presentados son los primeros que muestran el papel y la importancia del Cryptosporidium sp., y del Rotavirus en la DNB en las explotaciones ganaderas de la Sabana de Bogotá y en Colombia.
Objective. To determine the infectious agents causally associated to Bovine Neonatal Diarrhea (BND) in calves younger than five weeks of age from mixed production and dairy herds of Sabana de Bogota. Materials and methods. Twenty one herds were conveniently selected, and 620 calves were followed from birth up to 5 weeks of age, fecal samples were collected from animals with clinical signs of diarrhea and from calves without diarrhea which were taken as matched controls. ELISA tests were performed to diagnose E. coli F5, Rotavirus, Coronavirus, Salmonella sp, and Cryptosporidium sp, Ritchie test to diagnose Giardia sp., and a modified Ziehl Neelsen staining for Cryptosporidium sp. The epidemiological association between agents and the appearance of diarrhea was evaluated using c2 test, followed by a logistic regression model (p<0.05). Results. The ELISA test showed that 51 (38.3%), 26 (19.7%), 10 (7.5%) and 1 (0.75%) samples were positive for Cryptosporidium sp., rotavirus, E coli F5 and coronavirus, respectively. Animals that were positive for Rotavirus through ELISA and Cryptosporidium sp., through a modified Ziehl Neelsen technique had 2.6 and 7.0 times more probability than other animals to present BND, respectively. Conclusions. The results presented are the first to show the importance of Rotavirus and Cryptosporidium sp., in BND in the Sabana de Bogotá herds and in Colombia.
Assuntos
Diarreia , Cryptosporidium , Ensaio de Imunoadsorção Enzimática , RotavirusRESUMO
Foi investigado o valor diagnóstico da resposta alérgica cutânea à tuberculina e do ELISA indireto, com antígeno recombinante MPB 70, em leitões experimentalmente sensibilizados, pela via intramuscular, com suspensões oleosas de M. bovis ou M. avium inativados. Foram utilizados 91 animais divididos em quatro grupos. Os grupos A e B, cada um com 25 indivíduos, grupos C e D com 21 e 20 indivíduos respectivamente, balanceando-se as características de raça, linhagem, faixa etária e sexo. [...] Nos exames histopatológicos, foi observado intenso infiltrado inflamatório linfocitário no local das reações intradérmicas dos animais testados com o PPD homologo ao tipo de micobactéria utilizada na suspensão oleosa sensibilizante. O ensaio de ELISA com antígeno, MPB 70 recombinante, foi capaz de revelar a presença de anticorpos contra o M. bovis, porém não revelou anticorpos para M. avium
The diagnostic value of the cutaneous allergic response to tuberculin and Indirect ELISA test was investigated using MPB 70 recombinant antigen, in piglets experimentally sensitized intramuscularly with the oily suspensions of inactivated M. bovis or M. avium. The ninety-one animals used were divided into four groups. The groups A and B were formed each with 25 individuals, and groups C and D, with 21 and 20 individuals, respectively, balancing the characteristics of race, ancestry, age and sex. [...] In histopathological examinations, intense lymphocytic inflammatory infiltrate were observed at the site of intradermal reactions of the animals tested with PPD homologous to the type of mycobacteria used in sensitizing oil suspension. The ELISA assay with MPB 70 recombinant antigen was able to reveal the presence of antibodies against M. bovis, but did not reveal antibodies to M. avium
Assuntos
Animais , Doenças dos Suínos/patologia , Infecções por Mycobacterium/diagnóstico , Infecções por Mycobacterium/veterinária , Tuberculose/diagnóstico , Tuberculose/veterinária , Proteínas Recombinantes/imunologia , Testes Cutâneos/veterináriaRESUMO
The present research investigated the presence of T. evansi antibodies in animals from the subregion of Nhecolandia, in the Pantanal Sul-mato-grossense, by means of an enzyme linked immunosorbent assay (ELISA) and indirect immunofluorescence antibody test (IFAT), and the pattern of polypeptide recognition by sera from experimentally and naturally infected hosts using Western blotting. Serum samples were obtained from bovines (n = 102), horses (n = 98), and dogs (n = 55), and from 32 free-ranging coatis (Nasua nasua). None of the bovines were found positive, while sera from 16 dogs (29 percent) and 23 horses (23.4 percent) were positive by ELISA. Sera from 8 coatis (25 percent) were found positive using IFAT. Western blotting revealed major polypeptides of T. evansi with molecular weight ranging from 74 to 38 kDa. The polypeptides of 66, 48-46, and 38 kDa were identified by sera from experimentally infected bovines, donkeys, dogs, and coatis. The 48-46 and 38 kDa bands were mainly recognized in chronic phase of infection. The antigen with apparent molecular weight of 66 kDa, revealed by antibodies from all experimental animals, was also recognized in sera of horses and dogs from the Pantanal. The 48-46 kDa polypeptide was identified by antibodies from all naturally infected animals and must be further evaluated for use in specific diagnosis of T. evansi infection.
O trabalho de pesquisa investigou a presença de anticorpos anti- T. evansi em animais da sub-região da Nhecolândia, no Pantanal sul-mato-grossense, pelo ensaio imunoenzimático (ELISA) e a reação de imunofluorescência indireta (RIFI). O reconhecimento de polipeptídeos de T. evansi foi realizado pela técnica de "Western blotting", utilizando soros de animais experimentalmente e naturalmente infectados. As amostras de soro foram obtidas de bovinos (n = 102), cavalos (n = 98) e cães (n = 55) e de 32 quatis de vida livre (Nasua nasua) do pantanal mato-grossense. Todos os soros dos bovinos foram negativos, enquanto soros de 16 cães (29 por cento) e 23 cavalos (23,4 por cento) foram positivos pelo ELISA. Soros de oito quatis (25 por cento) foram positivos pela RIFI. Pelo "Western-blotting" foi possível revelar polipeptídeos de T. evansi, com peso molecular variando de 74 a 38 kDa. Os polipeptídeos de 66, 48-46 e 38 kDa foram identificados por soros de bovinos, cavalos, cães e quatis experimentalmente infectados. As bandas de 48-46 e 38 kDa foram reconhecidas principalmente na fase crônica da infecção. O antígeno com peso molecular aparente de 66 kDa, revelado por anticorpos de todos os animais experimentais, também foi reconhecido por soros de cavalos e cães do Pantanal. O polipeptídeo de 48-46 kDa foi identificado por anticorpos de todos os animais naturalmente infectados, devendo ser avaliado para o diagnóstico específico da infecção pelo T. evansi.
Assuntos
Animais , Anticorpos Antiprotozoários/sangue , Antígenos de Protozoários/sangue , Bovinos/sangue , Cães/sangue , Cavalos/sangue , Guaxinins/sangue , Trypanosoma/classificação , Trypanosoma/imunologiaRESUMO
The present research investigated the presence of T. evansi antibodies in animals from the subregion of Nhecolandia, in the Pantanal Sul-mato-grossense, by means of an enzyme linked immunosorbent assay (ELISA) and indirect immunofluorescence antibody test (IFAT), and the pattern of polypeptide recognition by sera from experimentally and naturally infected hosts using Western blotting. Serum samples were obtained from bovines (n = 102), horses (n = 98), and dogs (n = 55), and from 32 free-ranging coatis (Nasua nasua). None of the bovines were found positive, while sera from 16 dogs (29%) and 23 horses (23.4%) were positive by ELISA. Sera from 8 coatis (25%) were found positive using IFAT. Western blotting revealed major polypeptides of T. evansi with molecular weight ranging from 74 to 38 kDa. The polypeptides of 66, 48-46, and 38 kDa were identified by sera from experimentally infected bovines, donkeys, dogs, and coatis. The 48-46 and 38 kDa bands were mainly recognized in chronic phase of infection. The antigen with apparent molecular weight of 66 kDa, revealed by antibodies from all experimental animals, was also recognized in sera of horses and dogs from the Pantanal. The 48-46 kDa polypeptide was identified by antibodies from all naturally infected animals and must be further evaluated for use in specific diagnosis of T. evansi infection.
O trabalho de pesquisa investigou a presença de anticorpos anti- T. evansi em animais da sub-região da Nhecolândia, no Pantanal sul-mato-grossense, pelo ensaio imunoenzimático (ELISA) e a reação de imunofluorescência indireta (RIFI). O reconhecimento de polipeptídeos de T. evansi foi realizado pela técnica de "Western blotting", utilizando soros de animais experimentalmente e naturalmente infectados. As amostras de soro foram obtidas de bovinos (n = 102), cavalos (n = 98) e cães (n = 55) e de 32 quatis de vida livre (Nasua nasua) do pantanal mato-grossense. Todos os soros dos bovinos foram negativos, enquanto soros de 16 cães (29%) e 23 cavalos (23,4%) foram positivos pelo ELISA. Soros de oito quatis (25%) foram positivos pela RIFI. Pelo "Western-blotting" foi possível revelar polipeptídeos de T. evansi, com peso molecular variando de 74 a 38 kDa. Os polipeptídeos de 66, 48-46 e 38 kDa foram identificados por soros de bovinos, cavalos, cães e quatis experimentalmente infectados. As bandas de 48-46 e 38 kDa foram reconhecidas principalmente na fase crônica da infecção. O antígeno com peso molecular aparente de 66 kDa, revelado por anticorpos de todos os animais experimentais, também foi reconhecido por soros de cavalos e cães do Pantanal. O polipeptídeo de 48-46 kDa foi identificado por anticorpos de todos os animais naturalmente infectados, devendo ser avaliado para o diagnóstico específico da infecção pelo T. evansi.
RESUMO
El virus de bronquitis infecciosa (IBV) causa una enfermedad altamente contagiosa, distribuida mundialmente, que conlleva graves pérdidas económicas. En algunas oportunidades se asocia con otras entidades como los virus de las enfermedades de Gumboro y de Newcastle, Mycoplasma gallisepticum y Escherichia coli. La alta variabilidad genética del virus ha generado una gran cantidad de cepas virales con diferentes cuadros clínicos. El objetivo del trabajo fue evaluar la dinámica de anticuerpos del IBV en aves vacunadas y no vacunadas contra IBV, alojadas en una explotación de pollo de engorde donde se detectó el agente por RT-PCR, en Fusagasugá, Colombia, y aves vacunadas en semiaislamiento en Bogotá. Para esto se organizaron 3 grupos de aves (Ross 308) de 1 día de edad (44 aves/grupo), las cuales fueron vacunadas con un virus vivo atenuado, cepa Massachusetts H120, y se evaluó la respuesta inmune a través de la prueba de Elisa. Desde el primer día hasta el día 24 de edad se observó una disminución progresiva de los títulos de anticuerpos en los tres grupos,aunque en las aves vacunadas y no vacunadas mantenidas en granja se observaron niveles de anticuerpos superiores al grupo en condiciones de semiaislamiento. A partir del día 28en las aves alojadas en campo se incrementaron levemente los títulos hasta final de ciclo. El leve aumento en el nivel de anticuerpos puede ser consecuencia de exposición al virusvacunal que generó reversión de patogenicidad, persistencia viral o una exposición tardía al virus de campo.
The infectious bronchitis virus (IBV) causes a highly contagious disease, spread worldwide,leading to serious economic losses. Sometimes the disease is associated with other entities such as infectious bursal disease virus, Newcastle disease virus, Mycoplasma gallisepticumand Escherichia coli. The highly genetic variability of the virus has generated a large number of viral strains with different clinical presentations. The objective was to assess the dynamics of the virus antibodies in birds vaccinated and not vaccinated against IBV, hosted en ona broiler farm where the agent was detected by RT-PCR in Fusagasuga, Colombia and vaccinated birds in semi-isolation conditions in Bogotá. To order this, 3 groups of birds(Ross 308) from 1 day of age (44 birds/group), which were vaccinated with a live attenuated virus strain Massachusetts H120, and the immune response was evaluated through the Elisa test. Since day 24 of age the birds showed a progressive decrease in antibody titers in all three groups, although in the vaccinated and unvaccinated birds kept at the farm were found higher levels of antibodies in the group of semi-isolation. Starting at day 28 in the birds housed in field, the antibodies titles rose slightly until the end of cycle. The slight increase in the level of antibodies may result from exposure to the virus vaccine generated a reversal of pathogenic viral persistence or a late exposure to field virus.
Assuntos
Animais , Colômbia , Galinhas , Ensaio de Imunoadsorção Enzimática , Sorologia , Vírus da Bronquite InfecciosaRESUMO
Setenta e seis bovinos, sem raça definida, jovens, de ambos os sexos, apresentaram lesões cutâneas multifocais nodulares, ulceradas e crostosas nas faces medial e lateral dos membros anteriores e posteriores, região ventral do pescoço, esterno e cauda. A doença ocorreu no verão e as lesões foram observadas nas regiões do corpo que ficavam grande tempo em contato com a água em canais de irrigação. Histologicamente observaram-se múltiplos granulomas e piogranulomas contendo escassas imagens negativas de hifas na área central, as quais foram melhor evidenciadas através da técnica de metenamina nitrato de prata de Grocott. O diagnóstico etiológico definitivo foi baseado na técnica de imuno-histoquímica com anticorpo policlonal anti-Pythium insidiosum. Adicionalmente, foi realizado o teste de ELISA indireto. Surtos de pitiose cutânea bovina são incomuns e, particularmente neste relato, todos os animais afetados tiveram cura espontânea das lesões dentro de duas a três semanas.(AU)
Seventy-six young mixed breed cattle of both sexes, presented multifocal ulcerated nodular cutaneous lesions localized in the medial and lateral aspects of fore and hindlimbs, ventral neck, sternum, and tail. The disease occurred during summer and lesions were observed on areas of the body which were in contact with water of irrigation channels for long periods. Histologically, there were multiple granulomas and pyogranulomas with few negative profiles of hyphae, which were better visualized throughout Grocott methenamine silver stain. Definitive etiologic diagnosis was based on immuno-histochemistry with anti-Pythium insidiosum polyclonal antibody. Additionally, an indirect ELISA test was performed. Bovine cutaneous pythiosis outbreaks are uncommon and, particularly as occurred in the cattle of this report, all affected animals had spontaneous healing within two to three weeks.(AU)
Assuntos
Animais , Bovinos , Pythium/isolamento & purificação , Pitiose/epidemiologia , Granuloma/epidemiologia , Dermatopatias , Ensaio de Imunoadsorção Enzimática/veterinária , Imuno-Histoquímica/veterináriaRESUMO
Los herpesvirus equinos tipos 1 y 4 (HVE-1 y HVE-4) son agentes de distribución mundial y causa de graves pérdidas económicas. La infección primaria por ambos virus se produce en el tracto respiratorio, progresa a través de la mucosa y puede alcanzar otros sistemas orgánicos ocasionando abortos en el último tercio de gestación, muerte perinatal y síndromes neurológicos poco específicos. El HVE-1 está asociado principalmente con abortos, mientras que el HVE-4 se asocia con enfermedad respiratoria. Los animales afectados se recuperan sin tratamiento, pero permanecen infectados toda la vida. En 2001 se reportó en Colombia un aislamiento de HVE, pero hasta la fecha no se conoce ningún estudio que confirme y determine el tipo de herpesvirus aislado. El objetivo del presente trabajo fue realizar un estudio serológico para determinar la presencia de los HVE-1 y HVE-4 en equinos clínicamente sanos, no vacunados contra HVE. Para lograr nuestro objetivo se tomaron 139 muestras de suero de equinos de dos regiones de Colombia (Antioquia y Meta), a partir de las cuales se realizó una prueba de ELISA indirecta, para detectar la presencia de anticuerpos dirigidos contra la glicoproteína G del HVE-1 y HVE-4. Se encontró una seropositividad para HVE-4 del 98.7 y 96.6% en Antioquia y Meta respectivamente; para HVE-1, la seropositividad fue del 18.8% en el departamento de Antioquia y 33.3% en departamento del Meta. Este es el primer estudio que reporta seropositividad en equinos clínicamente sanos no vacunados en Colombia, lo cual sugiere la presencia del virus y su establecimiento en la población equina de las regiones evaluadas, y posiblemente de otras zonas de Colombia.
Equine herpesviruses 1 and 4 (EHV-1 and EHV-4) are world wide spread viruses that cause significant economic losses. The primary infection of both viruses occurs in the upper respiratory tract, it can progress trough the mucose and can cause abortions in the last third of pregnancy, neonatal foal death, and non specific neurological syndromes. The EHV-1 is associated mainly whit abortion whereas EHV-4 is more related to respiratory disease. Infected animals � rapidly overcome clinical symptoms but then remaininfected all the life. In 2001 the first isolation of an EHV was reported in Colombia, but up-to-date, there are no reports that confirm and determinethe type of isolated virus. The aim of the present work was to perform a serologic study in order to determine the presence of HVE-1 and HVE-4, in clinically healthy non-vaccinated horses. Serum samples n = 139 were drawn from healthy animals attwo regions of Colombia (Antioquia and Meta department); an indirect ELISA test was performed to evaluate the presence of antibodies recognizihg to glycoprotein G of EHV-1 and EHV-4. We found that 98.7 and 96.6% of sera in Antioquia and Meta respectively were positive for EHV-4; the positivity for EHV-1 in Antioquia and Meta were18.8 and 33.3% respectively. This isthe first study thatreports presence of antibodies to EHV-1 and EHV-4 in clinically healthy non-vaccinated horses in Colombia. These results suggest the establishment of the virus in the equine population of the studied regions and possibly in other areas of the country.
O herpesvirus eqüino tipo-1 e -4 (HVE-1 e HVE-4) são os agentes de distribuição mundial e causam graves prejuízos econômicos. A infecção primária por ambos os vírus ocorre no trato respiratório, progredindo através da mucosa e podem atingir outros sistemas orgânicos causando abortos no último terço da gestação, morte perinatal e não-específica síndromes neurológicas. O HVE-1 é principalmente associado a abortos, enquanto o HVE-4 está associado a doenças respiratórias. Os animais afectados recuperar sem tratamento, mas permanecem infectados vida. Em 2001, foi relatada na Colômbia isolamento de HVE, mas até agora não teve conhecimento de qualquer estudo para confirmar e identificar o tipo de herpesvírus isolados. O objetivo deste estudo foi o de realizar um levantamento sorológico para determinar a presença de HVE-1 e HVE-4 em cavalos clinicamente saudável, não vacinadas contra o HVE. Para alcançar nosso objetivo teve 139 amostras de soro de cavalos em duas regiões da Colômbia (Antioquia e Meta), de onde foi feito um teste Elisa indireta, para detectar a presença de anticorpos dirigidos contra a glicoproteína G da HVE -1 e da HVE-4. Para HVE-4 houve uma soropositividade de 98.7 e de 96.6% em Antioquia e Meta, respectivamente, para HVE-1, soropositividade foi 18.8% no departamento de Antioquia e de 33.3% no departamento de Meta. Este é o primeiro estudo que relatou soropositividade em cavalos clinicamente saudáveis não vacinados, na Colômbia, o que sugere a presença de vírus e seu lugar na eqüina população das regiões avaliadas e, possivelmente, de outras partes da Colômbia.
Assuntos
Animais , Ensaio de Imunoadsorção Enzimática , Herpesvirus Equídeo 1RESUMO
The aim of this study was to estimate the frequency of human toxocariosis in a child population from Morrope district, Lambayeque, Peru. From October to December 2005, 182 school children (96 male and 86 female) were studied. Blood samples were collected for Toxocara ELISA-IgG test and hematological examination. Additionally, stool samples were collected for coproparasitological examination to check cross reactions. We found frequency of positives in 32.4 percent (59/182) with a significant higher proportion of positivity in male children (p < 0.00001). 71.2 percent of the children with positive serology (52 male and seven female), were between five and 10 years old, 77.96 percent had respiratory symptoms, 61.02 percent had ocular manifestations, 38.98 percent had hepatic symptoms, 38.98 percent had mild or moderate eosinophilia, signs statistically associated with seropositivity. 83.5 percent of studied population had some intestinal parasite, such as: Blastocystis hominis (53.3 percent), Giardia lamblia (31.3 percent), Entamoeba coli (29.1 percent), Entamoeba histolytica/E. dispar (1.1 percent), Hymenolepis nana (5.49 percent), and Ascaris lumbricoides (3.3 percent), but they had not any association with serology results. The ownership of dogs or/and cats were significantly associated with seropositivity to anti-Toxocara antibodies although the presence of such pets within the house was not. In conclusion, clinical and serological evidence of Toxocara infection exists in the studied population.
O propósito do presente trabalho foi estimar a freqüência da toxocaríase humana em crianças do distrito de Morrope, Lamabayeque, Perú. Nos meses de outubro a dezembro de 2005, 182 escolares (96 meninos e 86 meninas) foram estudados. Coletaram-se amostras de sangue para a análise hematológica e imunológica pelo teste de ELISA-IgG-Toxocara e amostras fecais para o exame parasitológico. A freqüência geral da população foi de 32,4 por cento (59/182) com proporção significativamente maior de positividade nos meninos. Das crianças com resultado positivo ao teste sorológico (52 meninos e sete meninas), 71,2 por cento tinham entre cinco e 10 anos, 77,96 por cento apresentavam sintomas respiratórios, 61,02 por cento apresentavam manifestações oculares, 38,98 por cento apresentavam moléstias hepáticas, 38,98 por cento apresentavam suave ou moderada eosinofília e todos estes sinais foram estatisticamente associados ao resultado da sorologia. A presença de algum parasito intestinal foi observada em 83,5 por cento da população estudada: Blastocystis hominis (53,3 por cento), Giardia lamblia (31,3 por cento), Entamoeba coli (29,1 por cento), Entamoeba histolytica/E. dispar (1,1 por cento), Hymenolepis nana (5,49 por cento), e Ascaris lumbricoides (3,3 por cento), mas estes não tiveram nenhuma associação com a sorologia. A posse de cães e gatos revelou associação estatisticamente significante com a positividade para toxocaríase embora a presença destes animais no domicílio não tenha revelado tal associação. Conclui-se que existem evidências clínicas e sorológicas de infecção por Toxocara na população estudada.
Assuntos
Animais , Gatos , Criança , Pré-Escolar , Cães , Feminino , Humanos , Lactente , Masculino , Enteropatias Parasitárias/epidemiologia , Toxocara/isolamento & purificação , Toxocaríase/epidemiologia , Anticorpos Anti-Helmínticos/sangue , Ensaio de Imunoadsorção Enzimática , Fezes/parasitologia , Imunoglobulina G/sangue , Enteropatias Parasitárias/diagnóstico , Enteropatias Parasitárias/parasitologia , Prevalência , Peru/epidemiologia , Fatores de Risco , Índice de Gravidade de Doença , Toxocara/imunologia , Toxocaríase/diagnósticoRESUMO
O complexo teníase/cisticercose encontra-se universalmente distribuído, particularmente nos países em desenvolvimento e nos subdesenvolvidos. Neste, a precariedade das condições sanitárias e o baixo nível sócio-econômico cultural aliam-se na persistência de sua disseminação. Pontua-se ainda que, mesmo se fosse possível o exame de todas as massas musculares exploráveis nas condições normais do procedimento de inspeção de carnes, não seria possível assegurar-se, em caso de resultado negativo, que a carcaça estivesse livre de cisticercos. A presente pesquisa teve como objetivo identificar possíveis áreas endêmicas, através do teste sorológico em soro sanguíneo de suínos provenientes de criação doméstica no Estado do Mato Grosso. O material utilizado constituiu-se de 855 amostras de soro de suínos, onde se obteve uma freqüência de 34,4 por cento de reatividade para o teste de ELISA (Enzyme Linked Immunoabsorbent Assay).(AU)
The Taeniasis/Cysticercosis complex is disperse universally, privately in underdeveloped countries. In these, the uncertainty sanitaries conditions and the less level cultural socioeconomic to make an alliance in the persitence of your dissemination. To mention that, even if it is possible examinated all of muscles mass that con be explored in normal conditions of meat inspection procedement, wouldn't be possible to affirm, in negative result, that the carcass would be free of cysticerci. The present search had an objective to identified possible endemic areas, through the sorologic test in swine blood sera proceeding from domestic breeding of Mato Grosso State. The material used to as 855 swine sera sample, where obtained a reactivity frequency 34,4% for ELISA (Enzyme Linked Immuno absorbent Assay) Test. (AU)
Assuntos
Cisticercose/epidemiologia , Suínos/parasitologia , CarneRESUMO
An indirect ELISA (ENZYME-LINKED IMMUNOSORBENT ASSAY) was developed for searching of antibodies against-Cysticercus bovis in bovine. Three antigens were studied: partial antigen of C. cellulosae, total antigen of C. bovis, and total antigen of C. longicollis. In the standardization of the ELISA the following combinations were analyzed: antigen 250 and 500 ng of protein/well, dilution of the sera 50, 100, 200, and 400 times, dilution of the conjugated (anti bovine -IgG conjugated IgG of goat with peroxidase) 400 and 800 times. The crossing of the conditions above resulted in the following standardization: antigen 250 ng/well, sera and conjugated diluted 100 and 400 times respectively. The reaction cut-off between reagents and non-reagents animals was determined by the average of the optic densities of 54 negative sera plus three standard deviation resulting in the value of 0,303. The reactivity of the three antigens used in the ELISA test was compared using sera from experimentally infected calves, using sera dilutions and conjugated standardized previously. Using the antigen of C. bovis was verified high correlation with the test standardized with C. cellulosae. However, the absorbance values were significantly smaller. With C. longicollis was observed low reactivity, but increasing the amount of antigen, up to 3000ng/well, there was a proportional increase of the response. The kinetics of antibodies anti-C. bovis production was studied in ten calves experimentally infected with 2 x 104 T. saginata eggs. Six non-infected calves were used as control. After 90 days from the infection date, the animals were killed. Thirteen samples of sera of each animal were analyzed. The first was picked in the day of the infection and the remaining at each 15 days. The maximum production of antibody was observed between 30 and 60 days post infection. With the standardized test it was detected antibodies against-C. bovis, in 2 from 20 cattle considerated as non-holder of cyst by the inspection service. These animals could be considered possible cyst holders.
Um teste de ELISA indireto (ENZYME-LINKED IMMUNOSORBENT ASSAY) foi desenvolvido para pesquisa de anticorpos contra-C. bovis em bovinos experimental e naturalmente infectados. Foram estudados três antígenos: antígeno parcial de C. cellulosae, antígeno total de C. bovis e antígeno total de C. longicollis. Na padronização do ELISA foram analisadas as seguintes combinações: antígeno 250 e 500ng de proteína/cavidade, diluição dos soros 50, 100, 200 e 400 vezes, diluição do conjugado (IgG de cabra anti-IgG bovina conjugada com peroxidase) 400 e 800 vezes. Do cruzamento das condições acima resultou a seguinte padronização: antígeno 250ng/cavidade, soro e conjugado diluídos 100 e 400 vezes, respectivamente. O nível de corte (cut-off) da reação entre animais reagentes e não reagentes foi determinado pela média das densidades óticas de 54 soros negativos acrescidas de três desvio-padrão, resultando no valor de 0,303. Através da prova ELISA foram comparadas as reatividades dos antígenos parcial de C. cellulosae, total de C. bovis e total de C. longicollis com soros de bovinos portadores de cisticercose, empregando as diluições de soros e de conjugados padronizados anteriormente. O antígeno de C. bovis mostrou alta correlação com o teste padronizado com C. cellulosae. Entretanto, os valores de absorbância foram sensivelmente menores. Com C. longicollis observou-se reatividade bastante baixa, porém aumentando-se a quantidade de antígeno, até 3000ng/cavidade, houve um aumento proporcional da resposta. Após a padronização do teste foi analisado o comportamento imunológico de bezerros infectados experimentalmente com ovos de Taenia saginata. Dez bezerros foram infetados oralmente com 2 x 104 ovos de T. saginata. Seis bezerros não infetados foram usados como controle. Treze amostras de soro de cada animal foram analisadas. A primeira foi colhida no dia da infecção e o restante, quinzenalmente até o abate. A produção máxima de anticorpos foi observada entre 30 e 60 dias pós-infecção. Depois de 90 dias da infecção os animais foram sacrificados e o número de cistos contados e comparados com a resposta imunológica dos animais. Com o teste padronizado pesquisou-se anticorpos contra-C. bovis, em soros de bovinos considerados não portadores de cisticercos pelo serviço de inspeção e, de 20 amostras de soros analisadas, duas apresentaram valores de absorbância acima do "cut-off" indicando serem portadores de cisticercos .