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BACKGROUND: Accurate diagnosis of paracoccidioidomycosis is crucial for improving patient outcomes. Paracoccidioides antibody detection by double immunodiffusion (DID) is a convenient diagnostic tool, but testing performance can vary based on certain factors. METHODS: We assessed DID performance using a commercially prepared Paracoccidioides reagents (IMMY, USA), involving 40 serum specimens, including 20 from patients with proven paracoccidioidomycosis and 20 from patients without the disease. The DID test demonstrated a sensitivity of 90% (95% CI=68%-99%) and a specificity of 100% (95% CI=83%-100%). CONCLUSIONS: Our findings suggest that DID using commercial reagents may provide a feasible tool with satisfactory testing performance for anti-Paracoccidioides antibody detection.

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Definitive diagnosis of histoplasmosis relies on culture and/or cytology/histopathology; however, these procedures have limited sensitivity and cultures are time-consuming. Antibodies detection by immunodiffusion has low sensitivity in immunocompromised individuals and uses histoplasmin (HMN), a crude antigenic extract, as reagent. Novel protein antigen candidates have been recently identified and produced by DNA-recombinant techniques to obtain standardized and specific reagents for diagnosing histoplasmosis. To compare the analytical performance of novel enzyme-linked immunosorbent assays (ELISAs) for antibodies testing for diagnosing histoplasmosis using different Histoplasma capsulatum antigens as reagents. The H. capsulatum 100 kDa protein (Hcp100), the M antigen and its immunoreactive fragment F1 were produced by DNA-recombinant techniques. Galactomannan was purified from both the yeast and mycelial cell walls (yGM and mGM, respectively). The analytical performance of the ELISA tests for the serological detection of antibodies against these antigens was evaluated and compared with those obtained using HMN as reagent. Antibodies detection by the Hcp100 ELISA demonstrated 90.0% sensitivity and 92.0% specificity, versus 43.3% sensitivity and 95.0% specificity of the M ELISA, 33.3% sensitivity and 84.0% specificity of the F1 ELISA, 96.7% sensitivity and 94.0% specificity of the yGM ELISA, 83.3% sensitivity and 88.0% specificity of the mGM ELISA, and 70.0% sensitivity and 86.0% specificity for the HMN ELISA. In summary, Hcp100 is proposed as the most promising candidate for the serodiagnosis of histoplasmosis. The primary immunoreactive element in HMN proved to be GM rather than the M antigen. Nevertheless, a higher incidence of cross-reactions was noted with GM compared to M.

Hcp100 is a promising serodiagnostic candidate for histoplasmosis, boasting high sensitivity and specificity. Notably, GM, rather than M antigen, emerged as the primary immunoreactive element in HMN, despite a higher incidence of cross-reactions with GM compared to M.

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BACKGROUND: Equine infectious anaemia (EIA) is controlled by the identification of seropositive animals. The official diagnostic method is the agar gel immunodiffusion (AGID) test, which detects antibodies against a viral core protein (p26). Although AGID is inexpensive and specific, the report of results takes considerable time and the test has low analytical sensitivity. OBJECTIVE: To validate our in-house indirect ELISAgp90/45 , following the World Organization of Animal Health (OIE) criteria. STUDY DESIGN: Test validation. METHODS: Synthetic peptides gp90 and gp45 were used as antigens in ELISAgp90/45 . Tests used for validation, calibration and linear working operating range, analytical and diagnostic sensitivity and specificity, repeatability and reproducibility were assessed by comparing them with the AGID test and using 1844 equine sera grouped into five different panels. RESULTS: We were able to replace the National References Sera with our Internal Reference Sera. ELISAgp90/45 had acceptable repeatability and reproducibility. Analytical sensitivity of the ELISAgp90/45 was 800 times greater than that of AGID test for positive sera and 400 times greater for weak positive sera. ELISAgp90/45 also showed optimal analytical specificity, since no cross-reactivity was detected with antibodies against other equine viruses. One sample was positive by AGID test and negative by ELISAgp90/45. ELISAgp90/45 was performed using 243 EIA positive and 878 negative equid sera, and showed a diagnostic sensitivity of 99.59% [CI 97.73%-99.99%] and a diagnostic specificity of 90.32% [CI 88.17%-92.19%], compared to AGID test; thus, it was demonstrated to be a robust test. MAIN LIMITATIONS: Samples were derived from naturally infected equid populations showing heterogeneous clinical states: therefore, their status was uncertain and some horses were sampled more than once. The AGID test may not be the most useful gold standard. CONCLUSION: ELISAgp90/45 is a useful tool for the diagnosis of EIAV infection and meets validation requirements established by the OIE.

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Background: Serological evaluation performed by double agar gel immunodiffusion test (DID) is used for diagnosis, evaluation of severity, management of paracoccidioidomycosis patients, and development of new clinical studies. For these reasons, the Botucatu Medical School of UNESP maintains a serum bank at the Experimental Research Unit with patient clinical data. This study aimed to evaluate the influence of the freeze-thaw cycle and different blood matrices on the titration of circulating antibodies. Methods: The study included 207 patients with confirmed (etiology-demonstrated) or probable (serology-demonstrated) paracoccidioidomycosis, and DID was performed with culture filtrate from Paracoccidioides brasiliensis B339 as antigen. First experiment: the antibody levels were determined in serum samples from 160 patients with the chronic form and 20 with the acute/subacute form, stored at ­80o C for more than six months. Second experiment: titers of 81 samples of serum and plasma with ethylenediaminetetraacetic acid (EDTA) or heparin, from 27 patients, were compared according to matrix and effect of storage at ­20o C for up to six months. Differences of titers higher than one dilution were considered discordant. Results: First experiment: test and retest presented concordant results in serum stored for up to three years, and discordant titers in low incidence in storage for four to six years but high incidence when stored for more than six years, including conversion from reagent test to non-reagent retest. Second experiment: serum, plasma-EDTA and plasma-heparin samples showed concordant titers, presenting direct correlation, with no interference of storage for up to six months. Conclusions: Storage at ­80o C for up to six years has no or little influence on the serum titers determined by DID, permitting its safe use in studies depending on this parameter. The concordant titrations in different blood matrices demonstrated that the plasma can be used for immunodiffusion test in paracoccidioidomycosis, with stability for at least six months after storage at ­20o C.(AU)

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Small ruminant lentiviruses (SRLV) are difficult to diagnose due to their escape mechanisms. Therefore, proteomics is an alternative in the search for biomarkers through extracellular matrix metalloproteinases (MMPs), enzymes related to the immune response. In this sense, this study aimed to analyze the profile of MMPs in healthy and infected Toggenburg goats with chronic SRLV infection in Southeast Brazil. Five positive and five negative goats for SRLV were selected using the agar gel immunodiffusion (AGID) microtechnique, western blot (WB), and nested polymerase chain reaction (nPCR). All animals were submitted to blood collection by puncture of the jugular vein, followed by centrifugation to obtain blood plasma, protein quantification by the Bradford method, one-dimensional electrophoretic separation (1D), and identification of protease activity by zymography and confirmation via reverse zymography in the presence of MMP-2 through the action of tissue inhibitors (TIMP-2). The analysis of protein bands was performed using descriptive statistics and densitometry values for zymography were subjected to the Shapiro-Wilk test to determine normality. Little difference was observed in the occurrence of protein bands between groups. Regarding MMPs, no differences were observed in the expression of proMMP-9, MMP-9, and MMP-2 in animals affected by SRLV. TIMP-2 inhibited proMMP-2 and MMP-2 in all animals. Thus, the profile of protein bands does not change in healthy goats with chronic SRLV infection. The TIMP-2 expression allowed proving the existence of MMP-2 in animals chronically infected by SRLV via reverse zymography

Lentivírus de pequenos ruminantes (LVPR) demonstram diagnóstico complexo devido seus mecanismos de escape. Desse modo, a proteômica apresenta-se como alternativa na busca por biomarcadores através das metaloproteinases da matriz extracelular (MMPs), enzimas ligadas a resposta imunológica. Assim, objetivou-se analisar o perfil das MMPs em cabras Toggenburg sadias e com infecção crônica por LVPR no Sudeste brasileiro. Selecionou-se cinco cabras positivas e cinco negativas para LVPR utilizando: microtécnica de imunodifusão em gel de agarose (MIDGA), Western Blot (WB) e reação em cadeia da polimerase nested (nPCR). Todas foram submetidas à coleta de sangue por punção da veia jugular, seguido de centrifugação para obtenção do plasma sanguíneo, quantificação proteica pelo método Bradford, separação via eletroforese unidimensional (1D), e identificação da atividade das proteases por zimografia e confirmação via zimografia reversa na presença da MMP-2 por meio da ação de inibidores teciduais (TIMP-2). A análise das bandas proteicas ocorreu através de estatística descritiva e para a zimografia os valores de densitometria foram submetidos ao teste de Shapiro-Wilk para determinar a normalidade. Observou-se pouca distinção na ocorrência das bandas proteicas entre os grupos. Em relação as MMPs, não houve diferenças na expressão da proMMP-9, MMP-9 e MMP-2 nos acometidos por LVPR. Observou-se que a TIMP-2 inibiu a proMMP-2 e MMP-2 em todos os animais. Dessa forma, o perfil de bandas proteicas não se altera em cabras sadias e com infecção crônica por LVPR. Através da expressão da TIMP-2 foi possível comprovar a existência da MMP-2 em animais cronicamente infectados por LVPR via zimografia reversa

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The objective of this work was to describe the first record of antibodies to the Bluetongue Virus (BTV) in ewe, in the state of Amazonas. The ewe, which was in twin pregnancy, gave birth on May 9, 2015, but a lamb died hours after delivery. Veterinary service was then requested by the owner, where emaciation, loss of wool, pyrexia, apathy, dyspnea, mucoid nasal secretion, facial, lingual and submandibular edema were observed. There was a visit by the Agricultural Defense Agency of the State of Amazonas to the property and blood samples were collected from the animal. The whole blood and serum were sent to the National Agricultural Laboratory, where it was possible to detect the presence of specific antibodies to BTV, through the Agar Gel Double Immunodiffusion. The ewe was submitted to a new blood collection, following the same protocols and the samples were sent to the Biological Institute of São Paulo, confirmed diagnosis. The animal in a serious clinical condition, could not resist and died in July 2015. The occurrence of an allochthonous case, in an area where vector insects occur, can trigger an endemic process in the Amazon region. With this, the epidemiological control of these occurrences is necessary, in order to avoid the spread of the disease in the country.

O objetivo do trabalho foi descrever o primeiro registro de anticorpos para o Vírus da Língua Azul (VLA) em ovino, no estado do Amazonas. A ovelha, que se encontrava em gestação gemelar, pariu no dia 9 de maio de 2015, porém um cordeiro faleceu horas após o parto. Foi então solicitado serviço veterinário por parte do proprietário, onde foi observado emaciação, perda de lã, pirexia, apatia, dispneia, secreção nasal mucoide, edema facial, lingual e submandibular. Houve visita da Agência de Defesa Agropecuária do Estado do Amazonas na propriedade e coletadas amostras de sangue do animal. O sangue total e soro foram enviados ao Laboratório Nacional Agropecuário, no qual foi possível detectar a presença de anticorpos específicos para VLA, através do teste de Imunodifusão Dupla em Gel de Ágar. A ovelha foi submetida a uma nova coleta de sangue, seguindo os mesmos protocolos e as amostras foram enviadas ao Instituto Biológico de São Paulo, confirmando diagnóstico. O animal em estado clínico grave, não resistiu e veio a óbito em julho de 2015. A ocorrência de um caso alóctone, em uma área de ocorrência de insetos vetores, pode desencadear um processo de endemia na região amazônica. Com isso, o controle epidemiológico destas ocorrências, se fazem necessários, afim de se evitar a disseminação da doença no país.

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This study demonstrates the influence of pregnancy on serum diagnosis of enzootic bovine leukosis (EBL), emphasizing the importance of routine testing to maintain herd health. For this, 143 pregnant cows were sampled in duplicate (30 days before and 15 days after calving). For EBL diagnosis, samples were submitted to agar gel immunodiffusion testing (AGID). Different results were observed before and after delivery in seventy-six serum samples (53.15%), indicating variations in the levels of serum globulins in the blood during the peripartum period. Therefore, using a single sample for serological diagnosis during the birth season might not represent the correct infection status of animal health due to physiological variations in antibody concentrations.

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Background: Blue tongue (BT) is a noncontagious viral disease transmitted by hematophagous arthropods, especially of the genus Culicoides. The economic impact of the disease is related not only to deaths in sheep herds but also to the possible correlation of virus infection with the development of other diseases, such as pneumonia, abortion and movement problems. The economic losses caused by Blue Tongue are linked to restrictions on the import and export of animals and their genetic material and to the reproductive disorders associated with this disease. In addition, the fact that cattle take the role of reservoir, combined with the care by other countries with outbreaks of infection and biological contamination of their products, hinders trade in Mercosul, United States and Europe. Cattle are affected by Blue Tongue Virus in endemic areas and in some epidemic areas, but the development of clinical disease is rare. The clinical signs, when evident, range from reproductive losses, such as embryonic death, abortion, fetal malformation, temporary sterility, infertility in bulls, stillbirths and the birth of weak animals. The objective of this study was to determine the epidemiological aspects of Blue Tongue Virus (BTV) infection in dairy cattle in the Lavras region, state of Minas Gerais, Brazil. Materials, Methods & Results: A cross-sectional study was conducted to evaluate the frequency of cattle and herds seropositive for Blue Tongue in the southern region of Minas Gerais. In this study, 54 dairy farms were visited. A total of 586 serum samples were collected from cows of reproductive age. Sampling was random, and serum samples were collected from lactating cows over 24 months of age by puncture of the jugular vein and/or coccidian vein. The samples were transported and stored at the Setor de Patologia Veterinária, at the Universidade Federal de Lavras (SPV-UFLA), where they were centrifuged, and the serum aliquots were obtained, transferred to microtubes and kept at -20°C until the serological tests were performed. The samples were tested with the agarose gel immunodiffusion test (AGID) for anti-blue tongue virus antibodies. The AGID test is more practical and is the main method used to identify Blue Tongue Virus seroprevalence in different ruminant species. They are considered important tools for epidemiological surveillance of the disease. A prevalence of 83.28% was observed among animals that were seropositive for Blue Tongue Virus (488/586; IC 95% = 80.0 - 86.21). In addition, 100% (54/54; IC 95% = 93.4 - 100.0) of the farms had at least 1 positive animal, with rates ranging from 45.45% to 100% within the herds and where 22.22% of the farms had rates of 100% of the animals being positive. Discussion: Blue Tongue is a disease known to affect domestic and wild ruminants in Brazil. However, there is a lack of more precise information about its epidemiology and occurrence in the country and of joint efforts of researchers, producers and the government to understand in detail both the biology of vectors and the viral biology of Blue Tongue Virus in Brazil. This is the first record of detection of anti-blue tongue virus antibodies in cattle in the southern region of Minas Gerais. The results suggest that Blue Tongue Virus is present in cattle in the study area.

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Brucella ovis, a non-zoonotic species, is the etiological agent of ovine brucellosis, an infectious disease of clinical or subclinical occurrence in sheep flocks. Until then, there is no serological study of anti-Brucella ovis antibodies in purebred sheep herds. This study aimed to determine the presence of anti-Brucella ovis antibodies in purebred sheep flocks with breeding purposes from Parana State. Blood samples from 728 animals, of which 563 were females and 165 males, between 8 and 56 months of age from the six major sheep producing mesoregions of Parana, were submitted to detection of anti-Brucella ovis antibodies by the Agar Gel Immunodiffusion technique using an antigen from the bacteria Brucella ovis (Reo 198). The results indicate the presence of this disease in purebred sheep from Parana State in a low occurrence of 0.27% (2/728). The only two positive animals were rams, Santa Inês breed, from the same flock in the East Center region of Parana, without clinical disease. In conclusion, Brucella ovis is present in purebred sheep in Parana State, Brazil, and this low occurrence may have occurred due to rigorous breeding systems that may contribute to reduce the transmission of this disease.(AU)

Brucella ovis, espécie não zoonótica, é o agente etiológico da brucelose ovina, doença infecciosa de ocorrência clínica ou subclínica. Atualmente, não existe estudo sorológico de anticorpos anti-Brucella ovis em rebanhos de ovinos puros de origem. Este estudo teve como objetivo determinar a presença de anticorpos anti-Brucella ovis em rebanhos ovinos de raça pura de origem, com fins reprodutivos do estado do Paraná. Amostras de sangue de 728 animais, sendo 563 fêmeas e 165 machos, entre oito e 56 meses de idade, pertencentes a seis principais mesorregiões produtoras de ovinos no Paraná, foram submetidas à detecção de anticorpos anti-Brucella ovis pela técnica de imunodifusão em ágar gel usando-se um antígeno da bactéria Brucella ovis (Reo 198). Os resultados indicam a presença da doença em ovinos puros de origem do estado do Paraná em baixa ocorrência de 0,27% (2/728). Os dois únicos animais positivos foram reprodutores da raça Santa Inês, do mesmo rebanho da região Centro Leste do Paraná, sem manifestação clínica. Em conclusão, Brucella ovis está presente em ovinos puros de origem no estado do Paraná, e essa baixa ocorrência pode ter ocorrido devido a sistemas rigorosos de criação, que podem contribuir para a redução da transmissão dessa doença.(AU)

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PURPOSE: This article shows reports of the clinical-epidemiological characteristics and serological screening in patients assisted by a reference center for PCM care, University Hospital Cassiano Antonio Moraes, Federal University of Espirito Santo, Brazil. METHODS: The patient's sera with PCM were analyzed by DID test at the beginning and the end treatment. Clinical and demographic data were also collected to characterize the sample. RESULTS: One hundred patients with a suspected diagnosis of PCM were evaluated. Serology by DID test was used as a screen in all patients. The test was positive for 79 patients (72 for Paracoccidioides brasiliensis and 7 for Paracoccidioides lutzii). Serology was negative in 21 sera, although all of them were diagnosed PCM by histopathologic or direct exam. Serological follow-up was performed during the treatment of all patients. After treatment, 58(58%) had negative serology and 33(33%) low levels of antibodies (≤ 1:16). CONCLUSION: Our results indicate the importance of the DID test for the screening and monitoring of PCM and that the incidence of P. lutzii might be greater than expected in areas where it is not the predominant PCM species. Therefore, this article may contribute to improving the knowledge and clinical management about this disease.

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PURPOSE: In-house Paracoccidioides spp. antigens are commonly used in the serological diagnosis of paracoccidioidomycosis (PCM). The sensitivity and specificity of a commercial Paracoccidioides spp. antigen was assessed for PCM serological testing. METHOD: Counterimmunoelectrophoresis and double immunodiffusion were used to evaluate the Paracoccidioides ID Antigen® reagent in sera from PCM cases and patients with other diseases. RESULTS: All active PCM sera (n=24) were reactive using counterimmunoelectrophoresis (sensitivity = 100%), including 11 cases of infection by P. brasiliensis sensu stricto and one by P. americana. Fifteen (88%) out of 17 sera from patients on treatment or cured were reactive, including one case of P. lutzii infection. One to three bands of antigen-antibody precipitate were observed on the agarose gel, with a predominance of two to three bands in the test with untreated PCM sera or at the beginning of antifungal therapy. All sera from patients with histoplasmosis (n=7), aspergillosis (n=5), and other diseases (n=27) tested negative (specificity = 100%). The overall sensitivity and specificity using the commercial antigen and double diffusion test were 75% and 100%, respectively. CONCLUSION: The commercial antigen performed satisfactorily and may contribute to the dissemination of the use of serological tests for the PCM diagnosis.

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Mycobacterium avium subesp. paratuberculosis (MAP) e o vírus da leucemia bovina (BLV) são agentes que causam grandes perdas econômicas nos rebanhos. O objetivo deste estudo foi avaliar a situação epidemiológica da paratuberculose bovina (PTB) e leucose enzoótica bovina (EBL) em rebanhos leiteiros de Lagoa Formosa, Minas Gerais, Brasil. Foram coletadas 236 amostras de sangue de vacas, as quais foram submetidas aos testes ELISA e imunodifusão em gel de ágar para detecção de anticorpos contra MAP e BLV. A soroprevalência de anticorpos contra MAP e BVL foi de 20% para os rebanhos e 6% para os animais e de 85% para os rebanhos e 50,42% para os animais, respectivamente. A presença dessas enfermidades deve servir como um alerta para os produtores e veterinários, para que concentrem maior atenção na implementação de medidas higiênico-sanitárias, incorporando elementos de vigilância com base nos riscos identificados no estudo.(AU)

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The objective of this study was to verify the occurrence of ovine brucellosis using Agar Gel Immunodiffusion (AGID) and Polymerase Chain Reaction (PCR) techniques, as well as to identify the main risk factors associated with infection in sheep flocks belonging to municipalities in the microregion from Teresina, PI, Brazil. A total of 100 urine and blood samples were collected from sheep aged 6 months or older. The urine samples were submitted to conventional PCR and the blood samples were examined by the AGID technique. Of the 100 blood samples, 17 (17%) were reactive to the AGID test. In conventional PCR of 100 urine samples, six (6%) were positive. Risk factors associated to infection by B. ovis included the rearing system (OR=0.19), feed management (OR=0.05), presence of dystotic births (OR=4.50), miscarriages (OR=3.75) and source of water offered to the animals (OR=0.19). Thus, it was concluded that it is possible to detect the occurrence of animals with ovine brucellosis since PCR is a reliable method to confirm infection. Furthermore, there are risk factors associated to infection by B. ovis in the municipalities studied.

Objetivou-se verificar a ocorrência da brucelose ovina através das técnicas de Imunodifusão em Gel de Ágar (IDGA) e Reação em Cadeia da Polimerase (PCR), bem como identificar os principais fatores de risco associados à infecção nos rebanhos ovinos pertencentes a municípios da microrregião de Teresina, PI, Brasil. Foram colhidas 100 amostras de urina e de sangue de ovinos com idade superior ou igual a seis meses. As amostras de urina foram submetidas a PCR convencional e as amostras de sangue à técnica de IDGA. Das 100 amostras de sangue 17 (17%) foram reagentes ao teste de IDGA. Já na PCR convencional das 100 amostras de urina, seis (6%) foram positivas. Ressalta-se que três animais foram positivos em ambos os testes. Como fatores associados à infecção por B. ovis, observou-se o tipo de sistema de criação (OR=0,19), o manejo alimentar (OR=0,05), presença de partos distócicos (OR=4,50), abortamentos (OR=3,75) e a fonte de água fornecida aos animais (OR=0,19). Assim, conclui-se que foi possível detectar a ocorrência de animais com brucelose ovina, uma vez que a PCR é um método confirmatório. Além disso, há fatores de risco associados à infecção por B. ovis nos municípios estudados.

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The objective of this study was to verify the occurrence of ovine brucellosis using Agar Gel Immunodiffusion (AGID) and Polymerase Chain Reaction (PCR) techniques, as well as to identify the main risk factors associated with infection in sheep flocks belonging to municipalities in the microregion from Teresina, PI, Brazil. A total of 100 urine and blood samples were collected from sheep aged 6 months or older. The urine samples were submitted to conventional PCR and the blood samples were examined by the AGID technique. Of the 100 blood samples, 17 (17%) were reactive to the AGID test. In conventional PCR of 100 urine samples, six (6%) were positive. Risk factors associated to infection by B. ovis included the rearing system (OR=0.19), feed management (OR=0.05), presence of dystotic births (OR=4.50), miscarriages (OR=3.75) and source of water offered to the animals (OR=0.19). Thus, it was concluded that it is possible to detect the occurrence of animals with ovine brucellosis since PCR is a reliable method to confirm infection. Furthermore, there are risk factors associated to infection by B. ovis in the municipalities studied.

Objetivou-se verificar a ocorrência da brucelose ovina através das técnicas de Imunodifusão em Gel de Ágar (IDGA) e Reação em Cadeia da Polimerase (PCR), bem como identificar os principais fatores de risco associados à infecção nos rebanhos ovinos pertencentes a municípios da microrregião de Teresina, PI, Brasil. Foram colhidas 100 amostras de urina e de sangue de ovinos com idade superior ou igual a seis meses. As amostras de urina foram submetidas a PCR convencional e as amostras de sangue à técnica de IDGA. Das 100 amostras de sangue 17 (17%) foram reagentes ao teste de IDGA. Já na PCR convencional das 100 amostras de urina, seis (6%) foram positivas. Ressalta-se que três animais foram positivos em ambos os testes. Como fatores associados à infecção por B. ovis, observou-se o tipo de sistema de criação (OR=0,19), o manejo alimentar (OR=0,05), presença de partos distócicos (OR=4,50), abortamentos (OR=3,75) e a fonte de água fornecida aos animais (OR=0,19). Assim, conclui-se que foi possível detectar a ocorrência de animais com brucelose ovina, uma vez que a PCR é um método confirmatório. Além disso, há fatores de risco associados à infecção por B. ovis nos municípios estudados.

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ABSTRACT Purpose: In-house Paracoccidioides spp. antigens are commonly used in the serological diagnosis of paracoccidioidomycosis (PCM). The sensitivity and specificity of a commercial Paracoccidioides spp. antigen was assessed for PCM serological testing. Method: Counterimmunoelectrophoresis and double immunodiffusion were used to evaluate the Paracoccidioides ID Antigen reagent in sera from PCM cases and patients with other diseases. Results: All active PCM sera (n=24) were reactive using counterimmunoelectrophoresis (sensitivity = 100%), including 11 cases of infection by P. brasiliensis sensu stricto and one by P. americana. Fifteen (88%) out of 17 sera from patients on treatment or cured were reactive, including one case of P. lutzii infection. One to three bands of antigen-antibody precipitate were observed on the agarose gel, with a predominance of two to three bands in the test with untreated PCM sera or at the beginning of antifungal therapy. All sera from patients with histoplasmosis (n=7), aspergillosis (n=5), and other diseases (n=27) tested negative (specificity = 100%). The overall sensitivity and specificity using the commercial antigen and double diffusion test were 75% and 100%, respectively. Conclusion: The commercial antigen performed satisfactorily and may contribute to the dissemination of the use of serological tests for the PCM diagnosis.

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The objective of this study was to verify the occurrence of ovine brucellosis using Agar Gel Immunodiffusion (AGID) and Polymerase Chain Reaction (PCR) techniques, as well as to identify the main risk factors associated with infection in sheep flocks belonging to municipalities in the microregion from Teresina, PI, Brazil. A total of 100 urine and blood samples were collected from sheep aged 6 months or older. The urine samples were submitted to conventional PCR and the blood samples were examined by the AGID technique. Of the 100 blood samples, 17 (17%) were reactive to the AGID test. In conventional PCR of 100 urine samples, six (6%) were positive. Risk factors associated to infection by B. ovis included the rearing system (OR=0.19), feed management (OR=0.05), presence of dystotic births (OR=4.50), miscarriages (OR=3.75) and source of water offered to the animals (OR=0.19). Thus, it was concluded that it is possible to detect the occurrence of animals with ovine brucellosis since PCR is a reliable method to confirm infection. Furthermore, there are risk factors associated to infection by B. ovis in the municipalities studied.(AU)

Objetivou-se verificar a ocorrência da brucelose ovina através das técnicas de Imunodifusão em Gel de Ágar (IDGA) e Reação em Cadeia da Polimerase (PCR), bem como identificar os principais fatores de risco associados à infecção nos rebanhos ovinos pertencentes a municípios da microrregião de Teresina, PI, Brasil. Foram colhidas 100 amostras de urina e de sangue de ovinos com idade superior ou igual a seis meses. As amostras de urina foram submetidas a PCR convencional e as amostras de sangue à técnica de IDGA. Das 100 amostras de sangue 17 (17%) foram reagentes ao teste de IDGA. Já na PCR convencional das 100 amostras de urina, seis (6%) foram positivas. Ressalta-se que três animais foram positivos em ambos os testes. Como fatores associados à infecção por B. ovis, observou-se o tipo de sistema de criação (OR=0,19), o manejo alimentar (OR=0,05), presença de partos distócicos (OR=4,50), abortamentos (OR=3,75) e a fonte de água fornecida aos animais (OR=0,19). Assim, conclui-se que foi possível detectar a ocorrência de animais com brucelose ovina, uma vez que a PCR é um método confirmatório. Além disso, há fatores de risco associados à infecção por B. ovis nos municípios estudados.(AU)

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Equine infectious anemia (EIA) is one of the most important diseases from the health and economic point of view for equidae breeding, as it does not have treatment and vaccines. The Brazilian Ministry of Agriculture, Livestock and Food Supply (MAPA) instituted mandatory sanitary measures that include the official diagnosis by the agar gel immunodiffusion (AGID) test and sacrifice of seropositive animals to control this disease. Seventy-two seronegative equines, challenged with different vaccines, were used to verify the occurrence of non-specific reactions in the AGID techniques. Five serological controls were performed one week after vaccination, at seven-day intervals. The results indicated that the use of vaccines in equines in a period that precedes the performance of laboratory tests for the diagnosis of EIA does not induce seroconversion. However, 11.11% of the equines vaccinated against influenza, encephalomyelitis, equine rhinopneumonitis, and tetanus, and 15.38% of those vaccinated against leptospirosis had non-specific negative reactions to AGID. In this study, there was a non-specific line in the AGID for EIA described by Ordinance No. 84/1992 by MAPA but already mentioned in the Normative Instruction 55 of 26 November 2018.

Anemia Infecciosa Equina é uma das enfermidades mais importantes sob o ponto de vista sanitário e econômico para a equideocultura, por não possuir tratamento e vacinas. Para controle desta doença, o Ministério da Agricultura, Pecuária e Abastecimento (MAPA) instituiu medidas sanitárias obrigatórias em todo território nacional que incluem o diagnóstico e sacrifício dos animais soropositivos. Para verificar a ocorrência de reações inespecíficas na técnica de IDGA utilizou-se 72 equinos soronegativos, desafiados com diferentes vacinas. Uma semana após a vacinação, realizou-se cinco controles sorológicos, em intervalos de sete dias. Os resultados indicaram que o uso de vacinas em equinos em período que antecede a realização de exames laboratoriais para diagnóstico de AIE, não induz a soroconversão. Entretanto, 11,11% dos equinos vacinados contra influenza, encefalomielite, rinopneumonite equina e tétano, e 15,38% dos que foram vacinados contra leptospirose apresentaram reações negativas inespecíficas ao IDGA. Neste estudo, verificou-se uma linha inespecífica no IDGA para AIE.

Assuntos

RESUMO

Equine infectious anemia (EIA) is one of the most important diseases from the health and economic point of view for equidae breeding, as it does not have treatment and vaccines. The Brazilian Ministry of Agriculture, Livestock and Food Supply (MAPA) instituted mandatory sanitary measures that include the official diagnosis by the agar gel immunodiffusion (AGID) test and sacrifice of seropositive animals to control this disease. Seventy-two seronegative equines, challenged with different vaccines, were used to verify the occurrence of non-specific reactions in the AGID techniques. Five serological controls were performed one week after vaccination, at seven-day intervals. The results indicated that the use of vaccines in equines in a period that precedes the performance of laboratory tests for the diagnosis of EIA does not induce seroconversion. However, 11.11% of the equines vaccinated against influenza, encephalomyelitis, equine rhinopneumonitis, and tetanus, and 15.38% of those vaccinated against leptospirosis had non-specific negative reactions to AGID. In this study, there was a non-specific line in the AGID for EIA described by Ordinance No. 84/1992 by MAPA but already mentioned in the Normative Instruction 55 of 26 November 2018.(AU)

Anemia Infecciosa Equina é uma das enfermidades mais importantes sob o ponto de vista sanitário e econômico para a equideocultura, por não possuir tratamento e vacinas. Para controle desta doença, o Ministério da Agricultura, Pecuária e Abastecimento (MAPA) instituiu medidas sanitárias obrigatórias em todo território nacional que incluem o diagnóstico e sacrifício dos animais soropositivos. Para verificar a ocorrência de reações inespecíficas na técnica de IDGA utilizou-se 72 equinos soronegativos, desafiados com diferentes vacinas. Uma semana após a vacinação, realizou-se cinco controles sorológicos, em intervalos de sete dias. Os resultados indicaram que o uso de vacinas em equinos em período que antecede a realização de exames laboratoriais para diagnóstico de AIE, não induz a soroconversão. Entretanto, 11,11% dos equinos vacinados contra influenza, encefalomielite, rinopneumonite equina e tétano, e 15,38% dos que foram vacinados contra leptospirose apresentaram reações negativas inespecíficas ao IDGA. Neste estudo, verificou-se uma linha inespecífica no IDGA para AIE.(AU)

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RESUMO

Lentivirosis of small ruminants (LVPR) are chronic and degenerative infectious diseases, caused by Lentivirus, associated with numerous losses such as: drop in meat and milk production, predisposition to secondary infections, expenses with veterinary assistance and, even, early disposal of animals. In the northern region of Brazil, the epidemiological situation is poorly understood. Thus, this study aimed to determine the seropositivity of sheep for Lentivirus in Porto Acre city, Western Amazon, Brazil. 122 blood samples from sheep were collected and as a diagnostic method, agarose gel immunodiffusion was used, using the p28 protein of the capsid as antigen. The seropositivity of the sheep to the test was 8.2% (10/122). In 80% (4/5) of the investigated properties, the presence of seropositive animals was detected. It is worth noting that the acquisition of small ruminants from other states likely represented a risk to sheep health in the municipality of Porto Acre, Western Amazon, Brazil. It is concluded that there is a need for more systematic investigations on the prevalence of LVPR in the state of Acre.

As lentiviroses de pequenos ruminantes (LVPR) são enfermidades infecciosas crônicas e degenerativas, causadas por Lentivírus, associadas a inúmeros prejuízos como: queda na produção de carne e leite, predisposição a infecções secundárias, gastos com assistência veterinária e, até mesmo, descarte precoce dos animais. Na região norte do Brasil, a situação epidemiológica é pouco elucidada. Objetivou-se, assim, por meio deste estudo, determinar a soropositividade de ovinos para Lentivírus no município de Porto Acre, Amazônia Ocidental, Brasil. Foram coletadas 122 amostras de sangue de ovinos e como método diagnóstico foi empregada a imunodifusão em gel de agarose, utilizando a proteína p28 do capsídeo como antígeno. A soropositividade dos ovinos ao teste foi de 8,2% (10/122). Em 80% (4/5) das propriedades investigadas, detectou-se a presença de animais soropositivos. É válido ressaltar ainda que a aquisição de pequenos ruminantes advindos de outros estados provavelmente representou um risco à sanidade ovina no município de Porto Acre, Amazônia Ocidental, Brasil. Conclui-se que existe a necessidade de mais investigações sistemáticas sobre a prevalência de LVPR no estado do Acre.

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RESUMO

En los procesos neuroinflamatorios se produce a nivel de líquido cefalorraquídeo una activación policlonal y poliespecífica. Esta activación se produce desde los primeros días y puede permanecer por períodos prolongados. Luego por mecanismos de apoptosis los clones que no responden directamente contra los agentes biológicos involucrados no proliferan. El Reibergrama permite saber si las inmunoglobulinas presentes en el líquido cefalorraquídeo se sintetizaron o no en el sistema nervioso central (SNC) y el Índice de Anticuerpo (IA) determina la especificidad de las mismas en caso de que exista síntesis intratecal. Con estas herramientas nos propusimos identificar la respuesta neuroinmunológica frente a agentes de la familia herpesvirus en pacientes pediátricos con proceso inflamatorio del SNC a partir de sus respectivos IA. Para lograr esto se cuantificaron los niveles de IgG y albúmina en suero y líquido cefalorraquídeo (LCR) mediante inmunodifusión radial simple y por ensayo inmunoenzimático, con lo cual se construyó el Reibergrama que permitió la selección de 85 pacientes pediátricos con síntesis intratecal de inmunoglobulinas, que se diferenciaron en cuatro grupos según sus edades. Mediante ensayo inmunoenzimático se cuantificaron los niveles de IgG específica contra citomegalovirus, virus varicela zoster y virus herpes simple, tanto en suero como en LCR y se determinó el IA específico. La respuesta contra los virus estudiados fue similar para los distintos grupos de edades, lo cual nos permite afirmar la exposición temprana a los mismos(AU)

In a neuroinflammatory process a polyclonal and poly-specific activation is produced in cerebrospinal fluid. This activation starts from the first days and may persist for a long time. The clones not related directly against the biological agent do not proliferate by apoptosis. Reibergram determine if part of the immunoglobulins content in cerebrospinal fluid belongs from the blood or it is synthesized in the central nervous system. Antibody index determines if the specific antibodies was synthesized intrathecally. By these tools it can be possible to identify the humoral immune response against some herpes virus in pediatric patients suffering from a central nervous system inflammatory process. Quantification of specific IgG against citomegalovirus, varicella zoster and herpes simplex virus in serum and cerebrospinal fluid was done by ELISA. Specific Antibody index against these viruses were similar for the different age groups, which confirm the early exposure of the population(AU)

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