Seroprevalence, spatial analysis and risk factors of infection with Neospora caninum in cattle in Brazil's northern Pantanal wetland / Soroprevalência, análise espacial e fatores de risco da infecção por Neospora caninum em bovinos no Pantanal norte do Brasil

Abstract Neosporosis is caused by an obligate intracellular protozoan, Neospora caninum . It is considered one of the most widespread and frequent causes of abortion in cattle worldwide. To evaluate the prevalence of anti-N. caninum antibodies and associated risk factors, serum samples were collected from 2,452 bovines at 262 farms in the northern Pantanal, state of Mato Grosso, Brazil. Each farmer was asked to fill out a questionnaire for subsequent epidemiological data analysis. Anti-N. caninum antibodies were detected by means of the indirect immunofluorescent assay (IFA), using a cut-off dilution of 1:100. The overall anti-N. caninum antibodies prevalence was 25.44% (Confidence Interval - CI 95%; 20.10%; 30.78%), and the anti-N. caninum antibodies prevalence per herd was 76.72% (CI 95%; 71.60%; 81.84%). The presence of dogs, occurrence of abortion in cows, and sale of cattle for breeding were statistically associated with seropositivity in herds, while the risk of females being seropositive for N. caninum was higher in animals ≤ 6-years-old and in the presence of dogs. A spatial analysis indicated that the relative risk of the disease is spatially constant and that the farms with the highest prevalence of anti-N. caninum antibodies are located south of the region under study.

Resumo Neosporose é causada por um protozoário intracelular obrigatório, Neospora caninum. É considerada uma das causas mais comuns e frequentes de aborto em bovinos em todo o mundo. Para avaliar a prevalência de anticorpos anti- N. caninum e fatores de risco associados, amostras de soro foram coletadas de 2.452 bovinos em 262 fazendas no Pantanal norte, estado de Mato Grosso, Brasil. Cada fazendeiro preencheu um questionário para posterior análise dos dados epidemiológicos. Anticorpos anti-N. caninum foram detectados por meio da reação de imunofluorescência indireta (RIFI), utilizando um ponto de corte de 1:100. A prevalência total de anticorpos anti-N. caninum foi de 25,44% (Intervalo de Confiança - IC 95%; 20,10%; 30,78%) e a prevalência por rebanho foi de 76,72% (IC 95%; 71,60%; 81,84%). A presença de cães, a ocorrência de abortamento em vacas e a venda de bovinos para reprodução estiveram estatisticamente associadas à soropositividade em rebanhos, enquanto, o risco de fêmeas serem soropositivas para N. caninum foi maior em animais com idade ≤ 6 anos e na presença de cães. A análise espacial indicou que o risco relativo da doença é espacialmente constante e que as fazendas com maior prevalência de anticorpos anti-N. caninum estão localizados ao sul da região em estudo.

Seroprevalence, spatial analysis and risk factors of infection with Neospora caninum in cattle in Brazils northern Pantanal wetland

Abstract Neosporosis is caused by an obligate intracellular protozoan, Neospora caninum . It is considered one of the most widespread and frequent causes of abortion in cattle worldwide. To evaluate the prevalence of anti-N. caninum antibodies and associated risk factors, serum samples were collected from 2,452 bovines at 262 farms in the northern Pantanal, state of Mato Grosso, Brazil. Each farmer was asked to fill out a questionnaire for subsequent epidemiological data analysis. Anti-N. caninum antibodies were detected by means of the indirect immunofluorescent assay (IFA), using a cut-off dilution of 1:100. The overall anti-N. caninum antibodies prevalence was 25.44% (Confidence Interval - CI 95%; 20.10%; 30.78%), and the anti-N. caninum antibodies prevalence per herd was 76.72% (CI 95%; 71.60%; 81.84%). The presence of dogs, occurrence of abortion in cows, and sale of cattle for breeding were statistically associated with seropositivity in herds, while the risk of females being seropositive for N. caninum was higher in animals 6-years-old and in the presence of dogs. A spatial analysis indicated that the relative risk of the disease is spatially constant and that the farms with the highest prevalence of anti-N. caninum antibodies are located south of the region under study.

Resumo Neosporose é causada por um protozoário intracelular obrigatório, Neospora caninum. É considerada uma das causas mais comuns e frequentes de aborto em bovinos em todo o mundo. Para avaliar a prevalência de anticorpos anti- N. caninum e fatores de risco associados, amostras de soro foram coletadas de 2.452 bovinos em 262 fazendas no Pantanal norte, estado de Mato Grosso, Brasil. Cada fazendeiro preencheu um questionário para posterior análise dos dados epidemiológicos. Anticorpos anti-N. caninum foram detectados por meio da reação de imunofluorescência indireta (RIFI), utilizando um ponto de corte de 1:100. A prevalência total de anticorpos anti-N. caninum foi de 25,44% (Intervalo de Confiança - IC 95%; 20,10%; 30,78%) e a prevalência por rebanho foi de 76,72% (IC 95%; 71,60%; 81,84%). A presença de cães, a ocorrência de abortamento em vacas e a venda de bovinos para reprodução estiveram estatisticamente associadas à soropositividade em rebanhos, enquanto, o risco de fêmeas serem soropositivas para N. caninum foi maior em animais com idade 6 anos e na presença de cães. A análise espacial indicou que o risco relativo da doença é espacialmente constante e que as fazendas com maior prevalência de anticorpos anti-N. caninum estão localizados ao sul da região em estudo.

IgG Avidity ELISA Test for Diagnosis of Acute Toxoplasmosis in Humans

Serum samples, 100 in the total number, were collected from different laboratories in Tehran, Iran and tested for anti-Toxoplasma specific IgG and IgM antibodies using indirect immunofluorescent antibody test (IFAT). Using the IgG (chronic) and IgM (acute) positive samples, the IgG avidity test was performed by ELISA in duplicate rows of 96-well microtiter plates. One row was washed with 6 M urea and the other with PBS (pH 7.2), then the avidity index (AI) was calculated. Sixteen out of 18 (88.9%) sera with acute toxoplasmosis showed low avidity levels (AI60). Six sera had borderline ranges of AI. The results showed that the IgG avidity test by ELISA could distinguish the acute and chronic stages of toxoplasmosis in humans.

IgG Avidity ELISA Test for Diagnosis of Acute Toxoplasmosis in Humans

Serum samples, 100 in the total number, were collected from different laboratories in Tehran, Iran and tested for anti-Toxoplasma specific IgG and IgM antibodies using indirect immunofluorescent antibody test (IFAT). Using the IgG (chronic) and IgM (acute) positive samples, the IgG avidity test was performed by ELISA in duplicate rows of 96-well microtiter plates. One row was washed with 6 M urea and the other with PBS (pH 7.2), then the avidity index (AI) was calculated. Sixteen out of 18 (88.9%) sera with acute toxoplasmosis showed low avidity levels (AI60). Six sera had borderline ranges of AI. The results showed that the IgG avidity test by ELISA could distinguish the acute and chronic stages of toxoplasmosis in humans.

Detecção de anticorpos anti-Leishmania infantum syn chagasi em cães de São José do Rio Preto, São Paulo / Anti-Leishmania infantum syn chagasi antibodies in dogs from São José do Rio Preto, São paulo

A leishmaniose visceral (LV) é uma zoonose que vem se expandido por todo o território paulista desde 1998, quando foi identificado o primeiro caso canino autóctone, no município de Araçatuba. O objetivo do presente estudo foi determinar a ocorrência de anticorpos anti-Leishmania infantum syn chagasi em amostras de soro de 584 cães de São José do Rio Preto, São Paulo, área não endêmica para a doença. Cinco cães (0,86%) foram soro reagentes pela técnica de ELISA e um (0,17%) por imunocromatografia. A reação de imunofluorescência indireta, realizada em 138 animais que possuíam densidades ópticas acima ou próximas ao ponto de corte do ELISA evidenciou dois cães (1,45%) com títulos acima de 1:40. Apenas um animal foi sororeagente nas três técnicas sorológicas. Apesar deste cão não apresentar histórico de deslocamento para áreas endêmicas, havia sido adquirido em região com casos caninos e humanos de LV. Os resultados obtidos no presente estudo fazem supor que não existiam casos autóctones de LV canina na população estudada.

Visceral leishmaniasis (VL) has been a widespread zoonosis in São Paulo since 1998, when the first autochthonous canine case was identified in Araçatuba. The aim of this study was to determine the occurrence of anti–Leishmania infantum syn chagasi antibodies in serum samples of 584 dogs from São José do Rio Preto, São Paulo, a non endemic area for the disease. Five dogs (0.86%) seroconverted by ELISA and one (0.17%) by immunochromatography. The indirect immunofluorescent reaction, carried out in 138 animals whose optical densities were above or close to ELISA’s cutt-off point, evidenced two dogs (1.45%) with titers above 1:40. Only one dog was serum-reactive on the three techniques. Although there was not a history of displacing this animal to endemic areas, the dog had been acquired in a region with canine and human cases of VL. These results suggests that there were no autochthonous cases of canine VL in this population.

Experiência da adoção do I e II Consensos Brasileiros de Fator Antinuclear por Imunofluorescência Indireta em Células HEp-2 em um hospital universitário / Experience of an university hospital on the implementation of I and II Brazilian Consensuses for Standardization of ANA in HEp-2 Cells

OBJETIVO: Analisar a prevalência, padrões e títulos do Fator Antinuclear (FAN) por imunofluorescência indireta (IFI) em células HEp-2 em um hospital universitário após a adoção do I e II Consensos Nacional para Padronização dos Laudos de FAN em Células HEp-2. MÉTODO: Foi realizado um estudo transversal, em que foram revisados os laudos de FAN por IFI originários de solicitações encaminhadas ao Serviço de Patologia Clínica do Hospital de Clínicas de Porto Alegre (SPC/HCPA) entre 2002 e 2005. RESULTADOS: Foram analisados 12.095 testes de FAN no período entre 2002 e 2005. As solicitações com resultado reagente foram de 2.577 (21,30 por cento), com média anual de 644±233). Houve um aumento significativo na proporção de resultados reagentes posterior à adoção dos Consensos (p < 0,001). A Reumatologia foi a especialidade que mais solicitou exames por paciente atendido, mas houve um declínio nesse número nos anos posteriores à adoção do Consenso, ocorrido em 2004 (p < 0,001). O padrão de imunofluorescência de FAN mais frequentemente encontrado foi o padrão nuclear pontilhado fino, presente em 52,3 por cento dos resultados reagentes (453/866), e os títulos mais encontrados foram 1/80 e 1/160 (27,8 por cento e 29,4 por cento, respectivamente). CONCLUSÃO: Após a adoção do Consenso Nacional de Padronização de Laudos de FAN, observou-se um aumento de exames com resultados reagentes, na sua maioria, com títulos baixos e padrão nuclear pontilhado fino. Na Reumatologia, observou-se uma diminuição no número de solicitações desse exame. As potenciais causas para essas observações são discutidas, mas seu real impacto sobre a situação clínica do paciente e seu tratamento merece ser mais bem estudado.

OBJECTIVE: To evaluate the prevalence of patterns and titers of antinuclear antibodies (ANA) detected by indirect immunofluorescence (IIF) technique on HEp-2 cells in a university hospital following the introduction of I and II Brazilian Consensuses for Standardization of ANA in HEp-2 Cells. METHODS:A transversal study was performed between 2002 and 2005 during which all ANA orders to Serviço de Hospital de Clínicas de Porto Alegre (SPC/HCPA) and cognate results were reviewed. RESULTS:12.095 tests of ANA were revised. The number of positive results during this period was 2.577 (21.30 percent), annual mean 644 (SD: 233). A marked increase in the number of positive results was observed following the introduction of the Consensuses (p < 0.001). Rheumatology was the medical specialty which requested the highest number of ANA testing per patient although a significant decrease of these numbers was observed after the introduction of the Consensus in 2004 (p < 0,001). Nuclear fine speckled immunofluorescence labeling was the most frequently ANA pattern observed, 52.3 percent (453/866), and low ANA titers (1/80 and 1/160) more commonly detected (27.8 percent and 29.4 percent, respectively). CONCLUSION: Following the introduction of the Brazilian Consensus for standardization of ANA in HEp-2 cells an increased number of positive results was observed, mostly in low titers and with nuclear fine speckled immunofluorescence pattern. Moreover, there were decreasing numbers of ANA orders by rheumatologists in the same period. Potential causes for these observations are discussed but the real impact in the clinical condition of the patient and therapy deserves to be better studied.

Prevelance and Clinical Characteristics of Human Metapneumovirus in Hospitalized Children with Acute Respiratory Tract Infection / 实用儿科临床杂志

0.05).5.The clinical symptoms of hMPV infection could not be discriminated from the infection of other common respiratory viruses.Conclusion The acute respiratory-tract infections among children of Xi'an city are associated with cough and fever are major clinical symptoms.

Seroepidemiological Characteristics of Haemorrhagic Fever with Renal Syndrome from 1996 to 2005 in Korea

Haemorrhagic fever with renal syndrome (HFRS) caused by Hantaan viruses has been one of the principal acute febrile disease in Korea. To analysis the sero-epidemiological patterns of HFRS, 4,177 patient sera of acute febrile illness submitted for serological assay to National Institute of Health from Community Health Centers, Institutes of Health and Environment and hospitals from 1996 to 2005 were examined for antibodies against Hantaan virus by indirect immunofluorescent assay (IFA). Serum samples with greater than 1:32 antibody titer were considered positive. The results were analyzed seroepidemiologically by annual, sexual, seasonal, age and regional distribution of HFRS patients. Out of 4,177 serum samples tested, 1,415 samples (33.9%) were positive to Hantaan virus. The ratio of males (48.2%, 682/1,415) to females (38.2%, 541/1,415) was 1.3:1. Seasonal incidence showed that 69.5% (985/1,415) of cases occurred from October to December, resulting with higher prevalence in November (41.3%, 584/1,415). Regionally, seropositive rates of samples collected in Gyenggi, Gangwon and Chungbuk were 39.9% (564/1,415), 19.3% (274/1,415) and 8.5% (120/1,1415), respectively. Age distributions of seropositive of HFRS were detected from 20 to 79 years (78%).

Leishmaniasis tegumentaria: sensibilidad de la inmunofluorescencia indirecta en pacientes con diagnótico parasitológico / Tegumentary leishmaniasis: sensitivity of the indirect immunofluorescent assay in patients with parasitológical diagnosis

La leishmaniasis tegumentaria es una zoonosis endémica de zonas tropicales y subtropicales del mundo, de difícil diagnóstico laboratorial. En áreas endémicas se realiza el diagnóstico de presunción basado en la presentación clínica y la reactividad a la prueba cutánea de la leishmanina. Sin embargo, un diagnóstico definitivo de la enfermedad requiere la combinación de diferentes métodos, entre los cuales la inmunofluorescencia indirecta (IFI) es uno de los más utilizados. Para determinar la sensibilidad de la IFI, se sometió un total de 95 pacientes con lesiones comprobadas de leishmaniasis a las pruebas de leishmanina, biopsia histopatológica, cultivo e IFI. En 80 pacientes se logró el diagnóstico parasitológico a través de la histopatología y/o el cultivo. De ellos, 58 muestras (72,5%) dieron reacción positiva en la IFI para Leishmania, aunque 12 de ellas (21%) también mostraron serología positiva para enfermedad de Chagas. La sensibilidad de la IFI para los casos de leishmaniasis mucosa solamente fue 81,3% (n=16). Se concluye que la IFI es un método de moderada sensibilidad para el diagnóstico de leishmaniasis tegumentaria.

Tegumentary leishmaniasis is an endemic zoonosis of tropical and subtropical areas of the world. Diagnosis of this illness is difficult even using laboratorial methods. In endemics areas a presumptive diagnosis is carried out based on the clinical presentation and reactivity to the skin test. However, a definitive diagnosis of the illness requires the combination of different methods, such as the indirect immunofluorescence assay (IIF), one of the most used methods. In order to determine the sensitivity of the IIF, a total of 95 patients with proven lesions of leishmaniasis were subjected to skin test, histopathology, culture and IIF. Parasitological diagnosis was achieved in 80 patients through histopathology and/or culture. Fifty­eight samples of them (72.5%) gave positive reaction by the IIF although 12 of them (21%) also showed positive serology for Chagas disease. The sensitivity of the IIF for mucosal leishmaniasis only was 81.3% (n=16). We concluded that IIF has moderate sensitivity for the diagnosis of tegumentary leishmaniasis.

Clinical observation of patients with tsutsugamushi disease at Pyoungtaek area

BACKGROUND: The purpose of this stady is to define the clinical characteristics of Tsutsugamushi disease. METHODS: We analyzed 12 cases of tsutsugamushi disease admitted to BAG AE general hospital from 1999. Oct. 1st to Dec. 10. Tsutsugamushi disease was diagnosed by indirect immunofluorescent antibody test. RESULTS: 1) Among them, 8 cases were female and 4 cases were male. The prevalence was predominantly high in female. 2) The age distribution ranged from 40 to 80 years old. The high incidence of age was 70's and 80's. 3) The peak incidene was showed between October and November.(91.6%). 4) The most frequent symptoms were fever and chills(91.6%). 5) Rash and eschar were observed in 83.3%(10 of 12) both. 6) Hepatosplenomegaly and lymphadenopathy were also observed in 25%(3 of 12), 16.6%(2 of 12) respectively. 7) The Laboratory finding showed the elevation of AST 66.6% (8 of 12), ALT 58.3%(7 of 12), alkaline phosphatase 25%(3 of 12) and LDH 25%(3 of 12). Albuminuria and hypoalbuminuria were also observed in 25%(3 of 12), 58.3%(7 of 12) respectively. 8) The therapeutic efficacy of doxycycline was excellent. CONCLUSION: Tsutsugamushi disease which appeared in Pyoungtack area showed typical clinical pattern and was effective to doxycycline treatment.

Efficacy of Indirect Immunofluorescent Antibody Test in Herpes Simplex Keratitis

Herpes simplex virus keratits(HSK) is one of the most common external eye diseases that cause corneal blindness, Therefore early diagnosis and proper treatment of HSK are essential. However it is frequently misdiagnosed because it shows non-specific corneal lesion than other infectious corneal disease. And also diagnosis of HSK mostly rely on clinical examination and patient history. We evaluated suspicious HSK patients by indirect immunofluofluorescent(IF) antibody test and analyzed its efficacy in the early diagnosis of HSK. Among 47 patients(47 eyes), 37 patients were suspicious heretic keratitis and others not. Dendritic keratitis patients existed in 17 out of 37 patients and they were evaluated with virus culture and indirect IF test. The result of indirect IF test was confirmed under the immunofluorescent microscope and for virus culture the specimens were inoculated on Vero cells(monkey kidney cells). The positive results of indirect IF test was 28 out of 37 suspicious HSK patients and 1 out of 10 non-suspicious HSK patients. Dendritic HSK patients showed IF positive in 15 out of 17 patients(82.3%). Sensitivity of indirect IF test in suspicious HSK patients was 75.7%(2837) and 88.2%(15/17) in dendritic HSK patients. Indirect IF test was all positive(14/14) in dendritic HSK patients that showed culture positive. From these results, indirect IF test has a high sensitivity in early diagnosis of HSK and might be ussful as a rapid diagnostic tool in HSK patients.

Efficacy of Indirect Immunofluorescent Antibody Test in Herpes Simplex Keratitis

Herpes simplex virus keratits(HSK) is one of the most common external eye diseases that cause corneal blindness, Therefore early diagnosis and proper treatment of HSK are essential. However it is frequently misdiagnosed because it shows non-specific corneal lesion than other infectious corneal disease. And also diagnosis of HSK mostly rely on clinical examination and patient history. We evaluated suspicious HSK patients by indirect immunofluofluorescent(IF) antibody test and analyzed its efficacy in the early diagnosis of HSK. Among 47 patients(47 eyes), 37 patients were suspicious heretic keratitis and others not. Dendritic keratitis patients existed in 17 out of 37 patients and they were evaluated with virus culture and indirect IF test. The result of indirect IF test was confirmed under the immunofluorescent microscope and for virus culture the specimens were inoculated on Vero cells(monkey kidney cells). The positive results of indirect IF test was 28 out of 37 suspicious HSK patients and 1 out of 10 non-suspicious HSK patients. Dendritic HSK patients showed IF positive in 15 out of 17 patients(82.3%). Sensitivity of indirect IF test in suspicious HSK patients was 75.7%(2837) and 88.2%(15/17) in dendritic HSK patients. Indirect IF test was all positive(14/14) in dendritic HSK patients that showed culture positive. From these results, indirect IF test has a high sensitivity in early diagnosis of HSK and might be ussful as a rapid diagnostic tool in HSK patients.

A Study of Indirect Immunofluorescence Staining in the Diagnosis of Cutaneous Herpes Simplex Virus Infection

No abstract available.

A Clinical and Serologic Study of 21 Cases of Tsutsugamushi Disease Confirmed by Serologic test / 영남의대학술지

Tsutsugamushi disease is an acute febrile disease caused by Rickettsia tsutsugamushi, and which has been reported with increasing frequency thorough the nation since 1986. We experienced 21 cases of Tsutsugamushi Disease diagnosed with serologic test occurring in Taegu city and Kyungpook province during October-November, 1989. The results of survey are as follow. 1) Of 21 cases, 12 (57%) were males and 9 (43%) were females, and the peak incidence was the 4th decade. 2) The outbreak was in October to November and the peak incidence was in October. 3) The most frequent symptoms were fever and chill (100%), myalgia (95%), headache (90%), Eschar and rash were observed in 18 patients (86%) and the eschar was detected in all over the body, especially thorax (33%) and lower extremity (22%). 4) Laboratory features were SGOT elevation (83%), SGPT elevation (61%), LDH elevation (67%), leukocytosis (38%). 5) Indirect immunofluorescent antibody test was done in 18 patients and the antibody titer was above 1:320 in all patients. 6) The chloramphenicol, tetracycline or doxycycline regimens were very effective and mean duration of defervescence from initiation of therapy was 1.3 days. 7) The complication such as meningitis or shock, was not seen.

Immunofluorescent Studies of Various Chronic Bullous Dermatoses / 대한피부과학회지

The accuracy and sensitivity of both direct and indirect immunofluorescence microseopy in diagnosing chronic bullous dermatoses were evaluated and compared in 11 cases of six different disease entities(pemphigus vulgaris, pemphigus vegetans, bullous pemphigoid, linear IgA bullous dermatosis, familial benign chronic pemphigus), which had been diagnosed clinically and by routine histopa thological studies. And the results obtained were as follows: 1) In 4 cases of bullous pemphigod, the direct IF of perilesional skin showed linear deposition of IgG and C2 along basement membrane zone(BMZ), whereas the indirect IF revealed negetive findings. 2) In 2 cases of pemphigus vulgaris, the direct IF showed deposition of IgG and C2 in intercellular substance(ICS) of perilesional epidermal tissue, and the indirect IF revealed auto-antibody to ICS(1:320) in one case. 3) In 2 cases of linear IgA bullous dermatosis, which were diagnosed as dermatitis herpetiformis by clinical and routine hisopathological findings, the direct IF of perilesional and uninvolved skin manifested linear deposition of IgA, IgM, C3 and F along BMZ, However, no immunofluorescence was detected by the indirect IF. 4) In a case of chronic bullous dermatosis of childhood, the direct IF of perilesional skin showed linear deplosition of IgA and IgM along BMZ. 5) In a case of pemphigus vegetans, the direct IF revealed no specific findings, while the inderect IF disclosed auto antibody positive to ICS(1:40). 6) In a case of familial benign chronic permphigus, no immunofluorescence was found by direct IF. These results indicate that both direct and indirect immunofluorescence micros-copy are valuable in diagnosing chronic bullous dermatoses and in understanding their immune pathogenesis.

Construction of co-expression vector of pVAX1-PIG and expression of granzyme B and perforin in human Hep-2 cell line / 中国免疫学杂志

Objective:To amplify full-length expressing sequence of human granzyme B(GrB) and perforin(PFP) firstly,then to construct a co-expression vector of pVAX1-PIG and to analyze the expression of human GrB and PFP in Hep-2 cells.Methods:The human PFP and GrB expressing sequences were amplified by RT-PCR.The expression vector pVAX1-PIG was constructed.The recombinant vector was transfected into human Hep-2 cells and their expressions were detected by RT-PCR and indirect immunofluorescent antibody assay.Results:The GrB and PFP cDNA fragment were cloned into the vector of pVAX1 in the right direction.The target proteins were detected in the transfected Hep-2 cells.Conclusion:The vector of pVAX1-PIG was constructed successfully and expressed in Hep-2 cells line.The perforin/granzyme B protein can induce cell apoptosis.These results provide some foundation in gene therapy for tumor by making use of PFP and GrB gene.