Occurrence of Cryptosporidium in Anglonubian goats in the municipality of Teresina, state of Piauí, Brazil / Ocorrência de Cryptosporidium em cabras da raça Anglonubiana no município de Teresina, estado do Piauí, Brasil

This research aimed to investigate the occurrence of Cryptosporidium and correlate it with types of housing, feces consistency, and physiological parameters related to the reproductive status of Anglo-Nubian goats reared in the State of Piauí, Brazil. A total of 180 fecal samples were collected from 60 non-pregnant and lactating goats with a mean weight of 35 kg, a body condition score of 3.5, and a mean age of three years from an experimental herd at the Federal University of Piauí (UFPI). Oocysts of protozoa of the genus Cryptosporidium could be found in the studied animals using the modified Ziehl-Neelsen technique in fecal smears and the image analysis system to perform morphometry. Each independent variable in the quantitative and qualitative analyses, that is, weight, body condition score (BCS), physiological status (non-pregnant or lactating), feces consistency (normal, pasty, or diarrheal), and floor types (concrete and slatted), was tested with the dependent variable (positive samples, i.e., the presence of Cryptosporidium oocysts). Twenty-four out of the total number of fecal samples were considered positive for the presence of the protozoan, which means that 13.3% of the animals were parasitized. Moreover, 100% of the positive feces samples had normal consistency (firm) and all parasitized animals were reared in pens with a concrete floor. A statistical variation was observed in the BCS of parasitized animals compared to non-parasitized ones (p > 0.0253). The results showed that the occurrence of Cryptosporidium in experimental goats located in the municipality of Teresina, State of Piauí, Brazil, was considered low, requiring sanitary management measures to prevent infection in animals and humans. This is the first report of Cryptosporidium infection in goats in the State of Piauí.

O objetivo desta pesquisa foi investigar a ocorrência de Crypstoporidium e correlacionar com tipos de alojamento, consistência das fezes e parâmetros fisiológicos ligados ao estado reprodutivo de cabras da raça Anglonubiana criadas no estado do Piauí, Brasil. Foram utilizadas 180 amostras de fezes de 60 cabra, com peso médio de 35kg, escore de condição corporal de 3,5, com idade em média de três anos, e cabras vazias e lactantes, de um rebanho experimental da Universidade Federal do Piauí (UFPI). Utilizando-se a técnica de Ziehl-Neelsen modificada em esfregaço fecal e sistema de análise de imagens para a realização da morfometria, foi possível encontrar oocistos de protozoários do gênero Crypstoporidium nos animais estudados. Nas análises quantitativa e qualitativa, cada variável independente: peso, escore de condição corporal (ECC), estado fisiológico (vazia ou lactante), consistência das fezes (normal, pastosa ou diarreica) e tipos de piso (concreto e ripado), foi testada com a variável dependente (amostras positivas, ou seja, presença de oocistos de Cryptosporidium). Do total de amostras fecais analisadas, 24 delas foram consideradas positivas à presença do protozoário, o que significa que 13,3% dos animais estavam parasitados na ocasião da pesquisa. Foi observado que 100% das amostras de fezes positivas apresentaram consistência normal (firme) e que todos os animais parasitados eram criados em aprisco com piso de concreto. Houve uma variação estatística no ECC dos animais parasitados comparados aos não parasitados (p > 0,0253). Os resultados evidenciaram que a ocorrência de Cryptosporidium em caprinos experimentais localizados no município de Teresina, no estado do Piauí, foi considerada baixa, sendo necessária medidas de manejo sanitário para prevenir a infecção nos animais e no homem. Este é o primeiro relato da infecção por Cryptosporidium em cabras no estado do Piauí.

Cryptosporidium infection in diarrheal bovine dairy calves: occurrence and risk factors in Santa Catarina, Brazil / Infecção por Cryptosporidium em bezerros com diarreia: ocorrência e fatores de risco em Santa Catarina, Brasil

Cryptosporidium protozoa genus are parasites that cause acute enteric disease in young and immunocompromised animals, resulting in anorexia, loss and decrease in weight gain, and, in severe cases, death. Therefore, this study aimed: i) to determine the occurrence of Cryptosporidium spp. in calves with clinical diarrhea in different regions of Santa Catarina, Brazil; ii) to evaluate the risk factors involved with the frequency of infection. iii) to determine the species most involved with the disease in the region. For this, 425 samples were collected in 141 dairy farms, from animals with ages ranging from 0 to 150 days. For this purpose, the samples were submitted to the modified Ziehl-Neelsen technique, with molecular analysis of the positive samples being performed. It was observed 62.1% occurrence of Cryptosporidium spp. in this sampling, especially between 8 to 15 days. Regarding the risk factors evaluated, such as age, management, facilities, water source and Koppen climate (CFA and CFB), none showed statistical significance. Samples positive by the Ziehl-Neelsen technique (32 samples) were randomly selected for molecular diagnosis. Of these, 10 were sequenced, allowing the identification of Crypstosporidium parvum in 6 samples. However, this study proves the existence and high occurrence of the protozoan in different regions of the state of Santa Catarina, Brazil.

Os protozoários do gênero Cryptosporidium são parasitas que causam doença entérica aguda em animais jovens e imunocomprometidos, resultando em anorexia, perda e diminuição do ganho de peso e, em casos graves, morte. Portanto, este estudo teve como objetivo determinar a ocorrência de Cryptosporidium spp. em bezerros com diarreia clínica em diferentes regiões de Santa Catarina, Brasil; bem como avaliar os fatores de risco envolvidos com a frequência de infecção. Além disso, com um número seleto de amostras, buscou-se determinar as espécies mais envolvidas com a doença na região por meio de técnicas moleculares. Para isso, foram coletadas 425 amostras em 141 fazendas leiteiras, de animais com idade variando de 0 a 150 dias. Observou-se 62,1% de ocorrência de Cryptosporidium spp. nesta amostragem, principalmente entre 8 a 15 dias. Em relação aos fatores de risco avaliados, como idade, manejo, instalações, fonte hídrica e clima de Koppen (CFA e CFB), nenhum apresentou significância estatística. No entanto, este estudo comprova a existência e alta ocorrência do protozoário em diferentes regiões do estado de Santa Catarina, Brasil.

Comparison of different diagnostic modalities for isolation of Mycobacterium Tuberculosis among suspected tuberculous lymphadenitis patients / Comparação de diferentes modalidades de diagnóstico para isolamento de Mycobacterium tuberculosis entre pacientes com suspeita de linfadenite tuberculosa

Tuberculosis is a communicable disease with high morbidity and mortality rates in developing countries. The study's primary objective is to compare conventional methods such as acid-fast bacillus (AFB) culture and microscopy with rapid diagnostic methods. The secondary objective is to compare histopathological and microbiological findings in suspected patients with tubercular lymphadenitis. A total of 111 samples (August 2018 to September 2019) of lymph nodes were processed for AFB microscopy, AFB cultures, drug-susceptibility testing (DST), histopathology, and Xpert Mycobacterium Tuberculosis (MTB)/resistance to Rifampin (RIF) assays. Out of 111 lymph node samples, 6 (5.4%) were positive for AFB smear microscopy, 84 (75.6%) were positive for AFB culture, 80 (70.7%) were positive on Gene Xpert, and 102 (91.8%) were indicative of tuberculosis for histopathology studies. Mycobacteria growth indicator tube (MGIT) culture positivity was 84 (75.6%) higher than solid Lowenstein-Jensen (LJ) culture 74 (66.6%). Positive cultures underwent phenotypic DST. Two cases were Multidrug-resistant (MDR) on DST, while three cases were Rifampicin resistant on Gene Xpert. The sensitivity of Genexpert was (62%) against the conventional AFB culture method. The poor performance of conventional lymphadenitis diagnostic methods requires early and accurate diagnostic methodology. Xpert MTB/RIF test can help in the treatment of multidrug-resistant TB cases. Nonetheless, rapid and conventional methods should be used for complete isolation of Mycobacterium tuberculosis.

A tuberculose é uma doença transmissível com altas taxas de morbimortalidade nos países em desenvolvimento. O objetivo principal do estudo é comparar métodos convencionais, como cultura de bacilo álcool-ácido resistente (BAAR) e microscopia, com métodos de diagnóstico rápido. O objetivo secundário é comparar os achados histopatológicos e microbiológicos em pacientes com suspeita de linfadenite tubercular. Um total de 111 amostras (agosto de 2018 a setembro de 2019) de gânglios linfáticos foi processado para microscopia de AFB, culturas de AFB, teste de susceptibilidade a drogas (DST), histopatologia e Xpert Mycobacterium tuberculosis (MTB)/ensaios de resistência à rifampicina (RIF). Das 111 amostras de linfonodos, 6 (5,4%) foram positivas para baciloscopia de AFB, 84 (75,6%) foram positivas para cultura de AFB, 80 (70,7%) foram positivas para o GeneXpert e 102 (91,8%) foram indicativas de tuberculose para estudos histopatológicos. A positividade da cultura do tubo indicador de crescimento de micobactérias (MGIT) foi 84 (75,6%), maior que a cultura sólida de Lowenstein-Jensen (LJ), 74 (66,6%). As culturas positivas foram submetidas a DST fenotípico. Dois casos eram multirresistentes (MDR) ao DST, enquanto três casos eram resistentes à rifampicina no GeneXpert. A sensibilidade do GeneXpert foi 62% contra o método convencional de cultura AFB. O fraco desempenho dos métodos convencionais de diagnóstico de linfadenite requer metodologia de diagnóstico precoce e precisa. O teste Xpert MTB/RIF pode ajudar no tratamento de casos de tuberculose multirresistente. No entanto, métodos rápidos e convencionais devem ser usados para o isolamento completo do Mycobacterium tuberculosis.

Comparison of different diagnostic modalities for isolation of Mycobacterium Tuberculosis among suspected tuberculous lymphadenitis patients

Abstract Tuberculosis is a communicable disease with high morbidity and mortality rates in developing countries. The study's primary objective is to compare conventional methods such as acid-fast bacillus (AFB) culture and microscopy with rapid diagnostic methods. The secondary objective is to compare histopathological and microbiological findings in suspected patients with tubercular lymphadenitis. A total of 111 samples (August 2018 to September 2019) of lymph nodes were processed for AFB microscopy, AFB cultures, drug-susceptibility testing (DST), histopathology, and Xpert Mycobacterium Tuberculosis (MTB)/resistance to Rifampin (RIF) assays. Out of 111 lymph node samples, 6 (5.4%) were positive for AFB smear microscopy, 84 (75.6%) were positive for AFB culture, 80 (70.7%) were positive on Gene Xpert, and 102 (91.8%) were indicative of tuberculosis for histopathology studies. Mycobacteria growth indicator tube (MGIT) culture positivity was 84 (75.6%) higher than solid Lowenstein-Jensen (LJ) culture 74 (66.6%). Positive cultures underwent phenotypic DST. Two cases were Multidrug-resistant (MDR) on DST, while three cases were Rifampicin resistant on Gene Xpert. The sensitivity of Genexpert was (62%) against the conventional AFB culture method. The poor performance of conventional lymphadenitis diagnostic methods requires early and accurate diagnostic methodology. Xpert MTB/RIF test can help in the treatment of multidrug-resistant TB cases. Nonetheless, rapid and conventional methods should be used for complete isolation of Mycobacterium tuberculosis.

Resumo A tuberculose é uma doença transmissível com altas taxas de morbimortalidade nos países em desenvolvimento. O objetivo principal do estudo é comparar métodos convencionais, como cultura de bacilo álcool-ácido resistente (BAAR) e microscopia, com métodos de diagnóstico rápido. O objetivo secundário é comparar os achados histopatológicos e microbiológicos em pacientes com suspeita de linfadenite tubercular. Um total de 111 amostras (agosto de 2018 a setembro de 2019) de gânglios linfáticos foi processado para microscopia de AFB, culturas de AFB, teste de susceptibilidade a drogas (DST), histopatologia e Xpert Mycobacterium tuberculosis (MTB)/ensaios de resistência à rifampicina (RIF). Das 111 amostras de linfonodos, 6 (5,4%) foram positivas para baciloscopia de AFB, 84 (75,6%) foram positivas para cultura de AFB, 80 (70,7%) foram positivas para o GeneXpert e 102 (91,8%) foram indicativas de tuberculose para estudos histopatológicos. A positividade da cultura do tubo indicador de crescimento de micobactérias (MGIT) foi 84 (75,6%), maior que a cultura sólida de Lowenstein-Jensen (LJ), 74 (66,6%). As culturas positivas foram submetidas a DST fenotípico. Dois casos eram multirresistentes (MDR) ao DST, enquanto três casos eram resistentes à rifampicina no GeneXpert. A sensibilidade do GeneXpert foi 62% contra o método convencional de cultura AFB. O fraco desempenho dos métodos convencionais de diagnóstico de linfadenite requer metodologia de diagnóstico precoce e precisa. O teste Xpert MTB/RIF pode ajudar no tratamento de casos de tuberculose multirresistente. No entanto, métodos rápidos e convencionais devem ser usados para o isolamento completo do Mycobacterium tuberculosis.

Comparison of different diagnostic modalities for isolation of Mycobacterium Tuberculosis among suspected tuberculous lymphadenitis patients / Comparação de diferentes modalidades de diagnóstico para isolamento de Mycobacterium tuberculosis entre pacientes com suspeita de linfadenite tuberculosa

Abstract Tuberculosis is a communicable disease with high morbidity and mortality rates in developing countries. The study's primary objective is to compare conventional methods such as acid-fast bacillus (AFB) culture and microscopy with rapid diagnostic methods. The secondary objective is to compare histopathological and microbiological findings in suspected patients with tubercular lymphadenitis. A total of 111 samples (August 2018 to September 2019) of lymph nodes were processed for AFB microscopy, AFB cultures, drug-susceptibility testing (DST), histopathology, and Xpert Mycobacterium Tuberculosis (MTB)/resistance to Rifampin (RIF) assays. Out of 111 lymph node samples, 6 (5.4%) were positive for AFB smear microscopy, 84 (75.6%) were positive for AFB culture, 80 (70.7%) were positive on Gene Xpert, and 102 (91.8%) were indicative of tuberculosis for histopathology studies. Mycobacteria growth indicator tube (MGIT) culture positivity was 84 (75.6%) higher than solid Lowenstein-Jensen (LJ) culture 74 (66.6%). Positive cultures underwent phenotypic DST. Two cases were Multidrug-resistant (MDR) on DST, while three cases were Rifampicin resistant on Gene Xpert. The sensitivity of Genexpert was (62%) against the conventional AFB culture method. The poor performance of conventional lymphadenitis diagnostic methods requires early and accurate diagnostic methodology. Xpert MTB/RIF test can help in the treatment of multidrug-resistant TB cases. Nonetheless, rapid and conventional methods should be used for complete isolation of Mycobacterium tuberculosis.

Resumo A tuberculose é uma doença transmissível com altas taxas de morbimortalidade nos países em desenvolvimento. O objetivo principal do estudo é comparar métodos convencionais, como cultura de bacilo álcool-ácido resistente (BAAR) e microscopia, com métodos de diagnóstico rápido. O objetivo secundário é comparar os achados histopatológicos e microbiológicos em pacientes com suspeita de linfadenite tubercular. Um total de 111 amostras (agosto de 2018 a setembro de 2019) de gânglios linfáticos foi processado ​​para microscopia de AFB, culturas de AFB, teste de susceptibilidade a drogas (DST), histopatologia e Xpert Mycobacterium tuberculosis (MTB)/ensaios de resistência à rifampicina (RIF). Das 111 amostras de linfonodos, 6 (5,4%) foram positivas para baciloscopia de AFB, 84 (75,6%) foram positivas para cultura de AFB, 80 (70,7%) foram positivas para o GeneXpert e 102 (91,8%) foram indicativas de tuberculose para estudos histopatológicos. A positividade da cultura do tubo indicador de crescimento de micobactérias (MGIT) foi 84 (75,6%), maior que a cultura sólida de Lowenstein-Jensen (LJ), 74 (66,6%). As culturas positivas foram submetidas a DST fenotípico. Dois casos eram multirresistentes (MDR) ao DST, enquanto três casos eram resistentes à rifampicina no GeneXpert. A sensibilidade do GeneXpert foi 62% contra o método convencional de cultura AFB. O fraco desempenho dos métodos convencionais de diagnóstico de linfadenite requer metodologia de diagnóstico precoce e precisa. O teste Xpert MTB/RIF pode ajudar no tratamento de casos de tuberculose multirresistente. No entanto, métodos rápidos e convencionais devem ser usados ​​para o isolamento completo do Mycobacterium tuberculosis.

Comparison of different diagnostic modalities for isolation of Mycobacterium Tuberculosis among suspected tuberculous lymphadenitis patients / Comparação de diferentes modalidades de diagnóstico para isolamento de Mycobacterium tuberculosis entre pacientes com suspeita de linfadenite tuberculosa

Tuberculosis is a communicable disease with high morbidity and mortality rates in developing countries. The study's primary objective is to compare conventional methods such as acid-fast bacillus (AFB) culture and microscopy with rapid diagnostic methods. The secondary objective is to compare histopathological and microbiological findings in suspected patients with tubercular lymphadenitis. A total of 111 samples (August 2018 to September 2019) of lymph nodes were processed for AFB microscopy, AFB cultures, drug-susceptibility testing (DST), histopathology, and Xpert Mycobacterium Tuberculosis (MTB)/resistance to Rifampin (RIF) assays. Out of 111 lymph node samples, 6 (5.4%) were positive for AFB smear microscopy, 84 (75.6%) were positive for AFB culture, 80 (70.7%) were positive on Gene Xpert, and 102 (91.8%) were indicative of tuberculosis for histopathology studies. Mycobacteria growth indicator tube (MGIT) culture positivity was 84 (75.6%) higher than solid Lowenstein-Jensen (LJ) culture 74 (66.6%). Positive cultures underwent phenotypic DST. Two cases were Multidrug-resistant (MDR) on DST, while three cases were Rifampicin resistant on Gene Xpert. The sensitivity of Genexpert was (62%) against the conventional AFB culture method. The poor performance of conventional lymphadenitis diagnostic methods requires early and accurate diagnostic methodology. Xpert MTB/RIF test can help in the treatment of multidrug-resistant TB cases. Nonetheless, rapid and conventional methods should be used for complete isolation of Mycobacterium tuberculosis.(AU)

A tuberculose é uma doença transmissível com altas taxas de morbimortalidade nos países em desenvolvimento. O objetivo principal do estudo é comparar métodos convencionais, como cultura de bacilo álcool-ácido resistente (BAAR) e microscopia, com métodos de diagnóstico rápido. O objetivo secundário é comparar os achados histopatológicos e microbiológicos em pacientes com suspeita de linfadenite tubercular. Um total de 111 amostras (agosto de 2018 a setembro de 2019) de gânglios linfáticos foi processado para microscopia de AFB, culturas de AFB, teste de susceptibilidade a drogas (DST), histopatologia e Xpert Mycobacterium tuberculosis (MTB)/ensaios de resistência à rifampicina (RIF). Das 111 amostras de linfonodos, 6 (5,4%) foram positivas para baciloscopia de AFB, 84 (75,6%) foram positivas para cultura de AFB, 80 (70,7%) foram positivas para o GeneXpert e 102 (91,8%) foram indicativas de tuberculose para estudos histopatológicos. A positividade da cultura do tubo indicador de crescimento de micobactérias (MGIT) foi 84 (75,6%), maior que a cultura sólida de Lowenstein-Jensen (LJ), 74 (66,6%). As culturas positivas foram submetidas a DST fenotípico. Dois casos eram multirresistentes (MDR) ao DST, enquanto três casos eram resistentes à rifampicina no GeneXpert. A sensibilidade do GeneXpert foi 62% contra o método convencional de cultura AFB. O fraco desempenho dos métodos convencionais de diagnóstico de linfadenite requer metodologia de diagnóstico precoce e precisa. O teste Xpert MTB/RIF pode ajudar no tratamento de casos de tuberculose multirresistente. No entanto, métodos rápidos e convencionais devem ser usados para o isolamento completo do Mycobacterium tuberculosis.(AU)

Evaluation of fecal smear methods for research on Cryptosporidium spp. oocysts in the feces of dairy calves / Avaliação dos métodos de esfregaço fecal para a pesquisa de oocistos de Cryptosporidium spp. nas fezes de bezerros da bovinocultura de leite

The objective of this study is to compare the direct fecal smear (DFS) and centrifugal sedimentation (CS) methods in the detection of Cryptosporidium spp. oocysts in fecal samples of dairy calves. One hundred and fourteen fecal samples were collected from calves aged up to six months from 10 dairy farms located in Palotina and Francisco Alves, Paraná, Brazil. The microscopic analysis revealed the presence of Cryptosporidium spp. oocysts in 51.75% (59/114) of the samples in both methods. In CS, 48.25% (55/114) of the samples were positive, while in DFS slides, only 6.14% (7/114) were positive. Only 4 samples were positive exclusively in DFS. To ensure that there were no false-negative results in the microscopic analysis, the 55 samples that were negative in both DFS and CS were selected for molecular analysis using the nested PCR (nPCR). Of these 55 samples, 24% (13/55) were positive and forwarded for sequencing part of the genome, which made it possible to identify C. parvum, C. bovis and C. ryanae. Besides the characterization of the Cryptosporidium species, it was possible to identify bacteria of the genus Acinetobacter interfering directly in the analyzed samples. The microscopic analysis also revealed higher sensitivity when CS was used to make the fecal smears. However, some samples that were negative in this technique had positive PCR results. Thus, molecular analysis is indicated to confirm cases of Cryptosporidium spp. Further studies are necessary to prove the specificities of the used primers since the results obtained in nPCR were positive for the protozoan but, when genetic sequencing was performed, Acinetobacter spp. was identified.(AU)

O objetivo deste trabalho foi comparar os métodos de esfregaço fecal direto (DFS) e centrífugo sedimentação (CS) para a pesquisa de oocistos de Cryptosporidium spp. em amostras fecais de bezerros leiteiros. Foram coletadas 114 amostras fecais de bezerros com até seis meses de idade, provenientes de dez propriedades leiteiras localizadas nos municípios de Palotina e Francisco Alves, Paraná. Por meio da análise microscópica, foi possível observar oocistos de Cryptosporidium spp. em 51,75% (59/114) das amostras. Pelo método da CS foi identificada positividade em 48,25% (55/114) das amostras, enquanto nas lâminas pelo método do esfregaço fecal direto (DFS), apenas 6,14% (7/114) foram positivas. Somente 4 amostras foram positivas exclusivamente no método do DFS. Para assegurar que na análise microscópica não houvesse resultados falso-negativos, as 55 amostras negativas para os dois métodos de confecção de lâminas (DFS e CS) foram selecionadas para análise molecular por meio da técnica de Nested PCR. Das 55 amostras submetidas a nPCR, 24% (13/55) apresentaram-se positivas e foram encaminhadas para o sequenciamento genético de uma porção do genoma, o qual possibilitou identificar as espécies de C. parvum, C. bovis e C. ryanae. Além da caracterização das espécies de Cryptosporidium, foi possível identificar a presença de bactérias do gênero Acinetobacter interferindo diretamente nas amostras analisadas. A análise microscópica revelou maior sensibilidade quando o método de CS foi usado para a confecção dos esfregaços fecais, entretanto, amostras negativas por meio dessa metodologia apresentaram resultados positivos na nPCR. Desta forma, para a confirmação dos casos de Cryptosporidium spp., é indicado a realização da análise molecular. Novos estudos são necessários para se comprovar a especificidade dos primers utilizados, uma vez que na nPCR o resultado obtido foi positivo, porém ao realizar o sequenciamento genético houve a identificação de Acinetobacter spp.(AU)

Disseminated tuberculosis in a dog: a putative zooanthroponosis

Background: Zooantroponoses that affect pets are rare. Microorganisms involved in human tuberculosis has great importance to public health and has potentiality to produce reverse zoonoses. Infection by Mycobacterium tuberculosis is in top of the most important infectious diseases related to public health. In dogs, the infection occurs through prolonged exposure to aerosols and respiratory secretions from humans. Canine tuberculosis is little reported and known in Brazil, this work aims to describe the anatomopathological findings of the first case of canine tuberculosis in a 5-year-old dog in the Federal District and alert to the presence of this disease in this region.Case: The dog had history of close contact with its owner, who has been treated for pulmonary tuberculosis six months before. At necropsy scattered multifocal, firm, white to yellow-gray nodules, varying in size (3 mm to 10 mm) involving lungs, kidneys, heart, liver and spleen was observed. In frontal and occipital cerebral cortex, pale-gray circular lesions were detected with the same characteristics. The histological findings were, multifocal to coalescent granulomas, well organized with a central core of necrosis with foci of mineralization, surrounded by histiocytes, epithelioid macrophage, rare Langhans giant cells, some intact and degenerate plasma cells and neutrophils, and a rim of reactive fibroblasts producing fibrous connective tissue. Histological staining with ZN revealed free acid-fast bacilli and in the cytoplasm of macrophages and granulomas in all affected organs. The PCR allowed identification of DNA was extracted from formalin fixed paraffin embedded (FFPE) tissues using the ReliaPrep™ FFPE gDNA Miniprep System®, according to the manufacturer’s instructions (Promega, EUA). The Speed-Oligo Mycobacteria assay was used to identify the presence of M. tuberculosis complex DNA in tissue samples.[...]

Disseminated tuberculosis in a dog: a putative zooanthroponosis

Background: Zooantroponoses that affect pets are rare. Microorganisms involved in human tuberculosis has great importance to public health and has potentiality to produce reverse zoonoses. Infection by Mycobacterium tuberculosis is in top of the most important infectious diseases related to public health. In dogs, the infection occurs through prolonged exposure to aerosols and respiratory secretions from humans. Canine tuberculosis is little reported and known in Brazil, this work aims to describe the anatomopathological findings of the first case of canine tuberculosis in a 5-year-old dog in the Federal District and alert to the presence of this disease in this region.Case: The dog had history of close contact with its owner, who has been treated for pulmonary tuberculosis six months before. At necropsy scattered multifocal, firm, white to yellow-gray nodules, varying in size (3 mm to 10 mm) involving lungs, kidneys, heart, liver and spleen was observed. In frontal and occipital cerebral cortex, pale-gray circular lesions were detected with the same characteristics. The histological findings were, multifocal to coalescent granulomas, well organized with a central core of necrosis with foci of mineralization, surrounded by histiocytes, epithelioid macrophage, rare Langhans giant cells, some intact and degenerate plasma cells and neutrophils, and a rim of reactive fibroblasts producing fibrous connective tissue. Histological staining with ZN revealed free acid-fast bacilli and in the cytoplasm of macrophages and granulomas in all affected organs. The PCR allowed identification of DNA was extracted from formalin fixed paraffin embedded (FFPE) tissues using the ReliaPrep™ FFPE gDNA Miniprep System®, according to the manufacturers instructions (Promega, EUA). The Speed-Oligo Mycobacteria assay was used to identify the presence of M. tuberculosis complex DNA in tissue samples.[...](AU)

Tuberculosis of the central nervous system in cattle in Paraíba, Brazil / Tuberculose no sistema nervoso central de bovinos na Paraíba

This paper describes six cases of tuberculosis in the central nervous system (CNS) of cattle in the state of Paraíba in northeastern Brazil. We reviewed the autopsy reports of 851 bovine necropsies performed from 2003 to 2016. Seventy-three (8.6%) cattle were diagnosed with tuberculosis and six showed lesions in the CNS. Three cases affected cattle up to two-year-old and other three affected adults. Three cattle presented exclusively nervous signs, two had respiratory signs and weight loss and one did not present any clinical signs. At necropsy, five cattle had thickening of the leptomeninges of the cerebellum, pons, obex, spinal cord and cortex, mainly, in the region near the brain basilar Willis´ circle. Another animal, presented a single focal lesion in the cerebellum. Microscopically we observed moderate to severe granulomatous meningitis and encephalitis. Five cattle presented lesions in the lungs and mediastinal lymph nodes and three of them had disseminated lesions in other organs. In all cattle acid-fast bacilli were observed in the lesions and marked positive for immunohistochemistry with polyclonal antibody anti-Mycobacterium tuberculosis. It is concluded that bovine tuberculosis of central nervous system occurs sporadically in Paraíba, in cattle of different ages, most of them with disseminate lesions in other organs. The location of the lesions suggests that the agent invaded the brain by hematogenous route through the circle of Willis.(AU)

Descrevem-se seis casos de tuberculose no sistema nervoso central (SNC) em bovinos, no semiárido da Paraíba. Foram revisados os laudos de um total de 851 necropsias de bovinos realizadas no período de 2003 a 2016. Destes, 73 (8,6%) foram diagnosticados com tuberculose e seis apresentavam lesões no SNC. Três casos ocorreram em bovinos de até dois anos de idade e três em bovinos adultos. Três bovinos apresentaram exclusivamente sinais nervosos, dois tinham sinais respiratórios e perda de peso e um não apresentava nenhum sinal clínico. Macroscopicamente, em cinco bovinos, havia espessamento das leptomeninges do cerebelo, medula espinhal, ponte, obex, colículos e córtex, principalmente, na região basilar encefálica próxima ao polígono de Willis. Em apenas um bovino houve a presença de tubérculo único no cerebelo. Microscopicamente observou-se moderada a acentuada meningite e encefalite granulomatosa. Cinco bovinos apresentaram lesões pulmonares e nos gânglios mediastínicos e três deles tinham lesões disseminadas em outros órgãos. Em todos os bovinos foram encontrados bacilos ácido-álcool resistentes intralesionais e todos tiveram marcação positiva na técnica de imuno-histoquímica com anticorpo policlonal anti-Mycobacterium tuberculosis. Conclui-se que a tuberculose do sistema nervoso central de bovinos ocorre de forma esporádica na Paraíba, principalmente em bovinos com lesões disseminadas em outros órgãos. Sugere-se que a disseminação do agente ocorre pela via hematógena, possivelmente através do polígono de Willis.(AU)

Tuberculosis of the central nervous system in cattle in Paraíba, Brazil / Tuberculose no sistema nervoso central de bovinos na Paraíba

This paper describes six cases of tuberculosis in the central nervous system (CNS) of cattle in the state of Paraíba in northeastern Brazil. We reviewed the autopsy reports of 851 bovine necropsies performed from 2003 to 2016. Seventy-three (8.6%) cattle were diagnosed with tuberculosis and six showed lesions in the CNS. Three cases affected cattle up to two-year-old and other three affected adults. Three cattle presented exclusively nervous signs, two had respiratory signs and weight loss and one did not present any clinical signs. At necropsy, five cattle had thickening of the leptomeninges of the cerebellum, pons, obex, spinal cord and cortex, mainly, in the region near the brain basilar Willis´ circle. Another animal, presented a single focal lesion in the cerebellum. Microscopically we observed moderate to severe granulomatous meningitis and encephalitis. Five cattle presented lesions in the lungs and mediastinal lymph nodes and three of them had disseminated lesions in other organs. In all cattle acid-fast bacilli were observed in the lesions and marked positive for immunohistochemistry with polyclonal antibody anti-Mycobacterium tuberculosis. It is concluded that bovine tuberculosis of central nervous system occurs sporadically in Paraíba, in cattle of different ages, most of them with disseminate lesions in other organs. The location of the lesions suggests that the agent invaded the brain by hematogenous route through the circle of Willis.(AU)

Descrevem-se seis casos de tuberculose no sistema nervoso central (SNC) em bovinos, no semiárido da Paraíba. Foram revisados os laudos de um total de 851 necropsias de bovinos realizadas no período de 2003 a 2016. Destes, 73 (8,6%) foram diagnosticados com tuberculose e seis apresentavam lesões no SNC. Três casos ocorreram em bovinos de até dois anos de idade e três em bovinos adultos. Três bovinos apresentaram exclusivamente sinais nervosos, dois tinham sinais respiratórios e perda de peso e um não apresentava nenhum sinal clínico. Macroscopicamente, em cinco bovinos, havia espessamento das leptomeninges do cerebelo, medula espinhal, ponte, obex, colículos e córtex, principalmente, na região basilar encefálica próxima ao polígono de Willis. Em apenas um bovino houve a presença de tubérculo único no cerebelo. Microscopicamente observou-se moderada a acentuada meningite e encefalite granulomatosa. Cinco bovinos apresentaram lesões pulmonares e nos gânglios mediastínicos e três deles tinham lesões disseminadas em outros órgãos. Em todos os bovinos foram encontrados bacilos ácido-álcool resistentes intralesionais e todos tiveram marcação positiva na técnica de imuno-histoquímica com anticorpo policlonal anti-Mycobacterium tuberculosis. Conclui-se que a tuberculose do sistema nervoso central de bovinos ocorre de forma esporádica na Paraíba, principalmente em bovinos com lesões disseminadas em outros órgãos. Sugere-se que a disseminação do agente ocorre pela via hematógena, possivelmente através do polígono de Willis.(AU)

Occurrence of Cryptosporidium spp. oocytes in artificial beaches of Itaipu Lake, Paraná, Brazil / Ocorrência de oocistos de Cryptosporidium spp. em praias artificiais do Lago Itaipu, Paraná, Brasil

Cryptosporidiosis is an emerging zoonosis that causes many outbreaks worldwide. Its main route of transmission is the consumption of contaminated water or food. In Brazil, data on to the occurrence of cryptosporidiosis in animals and in humans are scarce; thus, it is important to determine the occurrence of Cryptosporidium sp. and to identify possible sources of water and food contamination in order to prevent the disease and, consequently, preserve of human health. From July/2011 to March/2012, five liters of water were collected in 10 different locations from eight artificial beaches of Itaipu Lake, Paraná, in a total of 40 samples. In two other small beaches of the lake, water was also collected from two different sites of each beach, given their size and because they were made up of water from different sources. Samples were analyzed by the modified Ziehl-Neelsen technique and nestedPCR (nPCR). All samples were negative in the modified Ziehl-Neelsen; only one (2.25%) sample was positive in nPCR. These results indicate that PCR is a more sensitive technique for the identification of Cryptosporidium in water samples. In this study, it was not possible to identify the species involved and, thus, the source of infection. Water contamination by Cryptosporidium oocytes can be caused by livestock kept near waterways, and by the people who use the place for recreational purposes. These results demonstrate risk for people who use the lake. Care should be determined to prevent the transmission of this important zoonosis.

A criptosporidiose é uma zoonose emergente responsável por vários surtos no mundo todo. Sua principal via de transmissão ocorre pelo consumo de água ou alimentos contaminados. Os dados de ocorrência de criptosporidiose tanto em animais como em seres humanos no Brasil são bastante escassos, dessa forma, torna-se importante a pesquisa da ocorrência de oocistos de Cryptosporidium sp. e a identificação de possíveis fontes de contaminação, visando a prevenção da contaminação da água e consequente preservação da saúde humana. Durante o período de jul/2011 a mar/2012 foram coletados cinco litros de água em 10 pontos de oito praias artificiais do Lago da Itaipu, Paraná, resultando em 40 amostras. Em duas prainhas foram coletadas água em dois locais devido ao tamanho das mesmas e por apresentarem fontes de água diferentes. As amostras foram analisadas pelo método de Ziehl-Neelsen modificado e nested-PCR. Das 40 amostras analisadas, todas foram negativas no método de ZiehlNeelsen modificado e uma (1) apresentou fragmento de 819-825 bp (2,25%) na técnica de nPCR, o que sugere que a PCR é uma técnica mais sensível para a pesquisa de Cryptosporidium em água. No presente trabalho não foi possível identificar a espécie envolvida, e dessa forma a fonte de infecção, ou espécie responsável pela contaminação da água não foi conhecida. A contaminação da água por oocistos de Cryptosporidium pode ser decorrente da criação animal nas proximidades dos cursos d’água e também pelas próprias pessoas que frequentam e utilizam o local. Esses resultados demonstram um risco para as pessoas que frequentam o local, devendo haver uma orientação quanto aos cuidados para evitar a transmissão dessa importante zoonose.

Occurrence of Cryptosporidium spp. oocytes in artificial beaches of Itaipu Lake, Paraná, Brazil / Ocorrência de oocistos de Cryptosporidium spp. em praias artificiais do Lago Itaipu, Paraná, Brasil

Cryptosporidiosis is an emerging zoonosis that causes many outbreaks worldwide. Its main route of transmission is the consumption of contaminated water or food. In Brazil, data on to the occurrence of cryptosporidiosis in animals and in humans are scarce; thus, it is important to determine the occurrence of Cryptosporidium sp. and to identify possible sources of water and food contamination in order to prevent the disease and, consequently, preserve of human health. From July/2011 to March/2012, five liters of water were collected in 10 different locations from eight artificial beaches of Itaipu Lake, Paraná, in a total of 40 samples. In two other small beaches of the lake, water was also collected from two different sites of each beach, given their size and because they were made up of water from different sources. Samples were analyzed by the modified Ziehl-Neelsen technique and nestedPCR (nPCR). All samples were negative in the modified Ziehl-Neelsen; only one (2.25%) sample was positive in nPCR. These results indicate that PCR is a more sensitive technique for the identification of Cryptosporidium in water samples. In this study, it was not possible to identify the species involved and, thus, the source of infection. Water contamination by Cryptosporidium oocytes can be caused by livestock kept near waterways, and by the people who use the place for recreational purposes. These results demonstrate risk for people who use the lake. Care should be determined to prevent the transmission of this important zoonosis.(AU)

A criptosporidiose é uma zoonose emergente responsável por vários surtos no mundo todo. Sua principal via de transmissão ocorre pelo consumo de água ou alimentos contaminados. Os dados de ocorrência de criptosporidiose tanto em animais como em seres humanos no Brasil são bastante escassos, dessa forma, torna-se importante a pesquisa da ocorrência de oocistos de Cryptosporidium sp. e a identificação de possíveis fontes de contaminação, visando a prevenção da contaminação da água e consequente preservação da saúde humana. Durante o período de jul/2011 a mar/2012 foram coletados cinco litros de água em 10 pontos de oito praias artificiais do Lago da Itaipu, Paraná, resultando em 40 amostras. Em duas prainhas foram coletadas água em dois locais devido ao tamanho das mesmas e por apresentarem fontes de água diferentes. As amostras foram analisadas pelo método de Ziehl-Neelsen modificado e nested-PCR. Das 40 amostras analisadas, todas foram negativas no método de ZiehlNeelsen modificado e uma (1) apresentou fragmento de 819-825 bp (2,25%) na técnica de nPCR, o que sugere que a PCR é uma técnica mais sensível para a pesquisa de Cryptosporidium em água. No presente trabalho não foi possível identificar a espécie envolvida, e dessa forma a fonte de infecção, ou espécie responsável pela contaminação da água não foi conhecida. A contaminação da água por oocistos de Cryptosporidium pode ser decorrente da criação animal nas proximidades dos cursos dágua e também pelas próprias pessoas que frequentam e utilizam o local. Esses resultados demonstram um risco para as pessoas que frequentam o local, devendo haver uma orientação quanto aos cuidados para evitar a transmissão dessa importante zoonose.(AU)

Occurrence and molecular characterization of Cryptosporidium sp. in sheep / Ocorrência e caracterização molecular de Cryptosporidium sp. em ovinos

Considered a zoonosis of utmost importance, cryptosporidiosis has a worldwide distribution and can infect mammals, birds, reptiles, and amphibians. It is caused by a highly resistant protozoan present in the environment and can cause death in immunosuppressed individuals and pups, as well as in farm animals such as cattle and sheep, generating losses. The aim of this study was to investigate the presence of Cryptosporidium spp. in sheep feces from the farms of Western Paraná, which have different management styles, and compare the results with their respective management methods. One hundred and forty-four stool samples were collected (69 from Property 1 and 75 from Property 2) and analyzed using a fecal smear on slides after staining by the modified Ziehl-Neelsen method. Samples tested positive by this method were subjected to nested PCR and the products obtained were sent for sequencing to determine the species. While 82.60% of the samples from Property 1 were tested positive, only 36% of the samples from Property 2 were tested positive. On analyzing the sequencing data, it was observed that the Cryptosporidium species of samples from Property 1 showed high similarity to Cryptosporidium xiaoi and those from Property 2, to Cryptosporidium ubiquitum. The reason for divergence in results can be attributed to differences in management systems adopted by each property, thus showing the importance of detecting carrier animals, as they can contaminate the environment,especially the water sources, and spread the disease to humans and other animals.(AU)

Considerada uma zoonose de extrema importância, a cryptosporidiose possui distribuição mundial e pode infectar mamíferos, aves, répteis e anfíbios. É causada por um protozoário muito resistente no ambiente, podendo gerar mortes em indivíduos imunossuprimidos e filhotes, principalmente em animais de produção como bovinos e ovinos, gerando prejuízos. O objetivo desse trabalho foi pesquisar a presença de Cryptosporidium spp. em fezes de ovinos de propriedades rurais do Oeste do Paraná, que possuem manejos diferenciados e comparar os resultados encontrados com o respectivo manejo. Foram coletadas 144 amostras de fezes, sendo 69 da Propriedade 1 e 75 da Propriedade 2, analisadas pelo esfregaço de fezes em lâminas após a coloração pelo método de Ziehl-Neelsen modificado. As amostras positivas por esse método foram submetidas à Nested-PCR sendo os produtos obtidos enviados ao sequenciamento para a determinação da espécie. A Propriedade 1 apresentou positividade em 82,60% das amostras, enquanto a Propriedade 2 obteve 36% de amostras positivas. Foi encontrada alta similaridade para Cryptosporidium xiaoi na primeira propriedade e Cryptosporidium ubiquitum na segunda propriedade. A divergência nos resultados pode ser comprovada pela diferente forma de manejo adotada em cada propriedade, mostrando a importância de detectar os animais portadores, uma vez que estes podem contaminar o meio ambiente, especialmente os cursos dágua, e promover a disseminaçãoda doença para outros animais e para o homem.(AU)

Occurrence of Cryptosporidium spp. in wild animals living in the Cascavel city park, Paraná, Brazil / Ocorrência de Cryptosporidium spp. em animais silvestres do Parque municipal de Cascavel, Paraná, Brasil

Digestive and respiratory tracts parasites protozoan, Cryptosporidium spp. now-a-days is a major zoonotic agent, it causes self-limiting diarrhea, remaining in the body passively until the moment immune system decreases, leading to an increase in its multiplication in the mucosa and appearance of clinical signs. As there are few studies on cryptosporidiosis in wild free-living and captive animals, especially in Brazil, this study aimed to identify the presence of Cryptosporidium spp. in feces of captive animals in Cascavel, PR Municipal Zoo. Between 2011 and 2012 there have been four collections of bird feces and five mammalian feces totaling 65 and 118 samples respectively. Samples were sent to the laboratory of Veterinary Parasitology of Federal University of Parana. The feces were diluted, centrifuged and the pellets were used to make blades which were stained by the Ziehl-Neelsen modified method and observed under a microscope with 1000X magnification. Then the blades containing the oocysts were observed under a capture microscope, where they were measured. Mammals showed 49.15% and birds 44.61% of positivity. Oocysts sizes ranged from 3,54?m to 5,81?m with an average of 4,32?m for birds and 3,11?m to 5,60?m averaging 4,63?m to mammals. As of yet, there isnt effective treatment against this parasite and considering that its a zoonotic disease, preventive measures should be taken to prevent transmission to humans.(AU)

Protozoário que parasita o trato digestório e respiratório dos animais, o Cryptosporidium spp. é um importante agente zoonótico da atualidade, responsável por causar diarreia auto limitante, permanecendo no organismo de forma passiva até o momento em que há uma queda da imunidade, levando a um aumento na sua multiplicação na mucosa e aparecimento de sinais clínicos. Como há poucos estudos sobre a criptosporidiose em animais silvestres de vida livre e de cativeiro, especialmente no Brasil, esse trabalho teve como objetivo realizar uma pesquisa para identificar a presença de oocistos de Cryptosporidium spp. em fezes dos animais cativos no Zoológico Municipal de Cascavel, Paraná. No período de 2011 e 2012 foram realizadas quatro coletas de fezes de aves e cinco de fezes de mamíferos totalizando 65 e 118 amostras respectivamente. As amostras foram enviadas ao laboratório de parasitologia veterinária da Universidade Federal do Paraná, Setor Palotina. As fezes foram diluídas, centrifugadas e os sedimentos foram utilizados para a confecção de lâminas, que foram coradas pelo método de Ziehl-Neelsen modificado e observadas em microscópio com aumento 1000X. Em seguida as lâminas contendo os oocistos foram observadas em microscópio de captura, onde os mesmos foram mensurados. Os mamíferos apresentaram 49,15% de positividade e as aves 44,61% das amostras positivas. O tamanho dos oocistos variou de 3,54μm a 5,81μm com uma média de 4,32μm para aves e de 3,11μm a 5,60μm com média de 4,63μm para mamíferos. Como ainda não há tratamento eficaz para esse parasito e considerando que se trata de uma zoonose, medidas preventivas devem ser tomadas para evitar a transmissão para humanos.(AU)

Occurrence of Cryptosporidium spp. in wild animals living in the Cascavel city park, Paraná, Brazil / Ocorrência de Cryptosporidium spp. em animais silvestres do Parque municipal de Cascavel, Paraná, Brasil

Digestive and respiratory tracts parasites protozoan, Cryptosporidium spp. now-a-days is a major zoonotic agent, it causes self-limiting diarrhea, remaining in the body passively until the moment immune system decreases, leading to an increase in its multiplication in the mucosa and appearance of clinical signs. As there are few studies on cryptosporidiosis in wild free-living and captive animals, especially in Brazil, this study aimed to identify the presence of Cryptosporidium spp. in feces of captive animals in Cascavel, PR Municipal Zoo. Between 2011 and 2012 there have been four collections of bird feces and five mammalian feces totaling 65 and 118 samples respectively. Samples were sent to the laboratory of Veterinary Parasitology of Federal University of Parana. The feces were diluted, centrifuged and the pellets were used to make blades which were stained by the Ziehl-Neelsen modified method and observed under a microscope with 1000X magnification. Then the blades containing the oocysts were observed under a capture microscope, where they were measured. Mammals showed 49.15% and birds 44.61% of positivity. Oocysts sizes ranged from 3,54?m to 5,81?m with an average of 4,32?m for birds and 3,11?m to 5,60?m averaging 4,63?m to mammals. As of yet, there isnt effective treatment against this parasite and considering that its a zoonotic disease, preventive measures should be taken to prevent transmission to humans.

Protozoário que parasita o trato digestório e respiratório dos animais, o Cryptosporidium spp. é um importante agente zoonótico da atualidade, responsável por causar diarreia auto limitante, permanecendo no organismo de forma passiva até o momento em que há uma queda da imunidade, levando a um aumento na sua multiplicação na mucosa e aparecimento de sinais clínicos. Como há poucos estudos sobre a criptosporidiose em animais silvestres de vida livre e de cativeiro, especialmente no Brasil, esse trabalho teve como objetivo realizar uma pesquisa para identificar a presença de oocistos de Cryptosporidium spp. em fezes dos animais cativos no Zoológico Municipal de Cascavel, Paraná. No período de 2011 e 2012 foram realizadas quatro coletas de fezes de aves e cinco de fezes de mamíferos totalizando 65 e 118 amostras respectivamente. As amostras foram enviadas ao laboratório de parasitologia veterinária da Universidade Federal do Paraná, Setor Palotina. As fezes foram diluídas, centrifugadas e os sedimentos foram utilizados para a confecção de lâminas, que foram coradas pelo método de Ziehl-Neelsen modificado e observadas em microscópio com aumento 1000X. Em seguida as lâminas contendo os oocistos foram observadas em microscópio de captura, onde os mesmos foram mensurados. Os mamíferos apresentaram 49,15% de positividade e as aves 44,61% das amostras positivas. O tamanho dos oocistos variou de 3,54μm a 5,81μm com uma média de 4,32μm para aves e de 3,11μm a 5,60μm com média de 4,63μm para mamíferos. Como ainda não há tratamento eficaz para esse parasito e considerando que se trata de uma zoonose, medidas preventivas devem ser tomadas para evitar a transmissão para humanos.

OCORRÊNCIA DE Cryptosporidium spp. E FATORES DE RISCO ASSOCIADOS À INFECÇÃO EM BEZERROS COM DIARREIA NO ESTADO DE SANTA CATARINA.

O Cryptosporidium spp. é um protozoário que tem se destacado por ser a causa mais comum de diarreia e atraso no crescimento em bezerros. Por ser considerado um patógeno oportunista, pode causar mortalidade, em geral, quando associado a outros agentes que causam patologias entéricas. A criptosporidiose acontece pela ingestão dos oocistos de Cryptosporidium spp. e a ocorrência está associada às condições higiênicas precárias, elevada densidade de bezerros em piquetes e a idade dos animais. O objetivo principal deste estudo foi determinar a prevalência de infecção por Cryptosporidium spp. em bezerros com o sinal clínico de diarreia, assim como avaliar os aspectos epidemiológicos associados à frequência da infecção. A amostragem por região foi baseada na população total, na margem de erro e no número de desvios padrões, selecionando bezerras com idade que variou do nascimento até 150 dias de idade, totalizando 425 amostras oriundas de 141 propriedades de diferentes regiões do estado de Santa Catarina. Para o diagnóstico, as amostras foram submetidas à técnica de Sheather seguida da coloração de Ziehl-Neelsen modificado. Observou-se uma prevalência de 62,1% de amostras positivas para o protozoário Cryptosporidium spp. nesses animais. Quando analisados alguns fatores associados à infecção pelo Cryptosporidium spp. em bezerras, pode-se observar que animais mantidos em instalações individuais apresentaram 69.5% dos casos positivos, e animais que consumiam água de vertente foram os mais acometidos, com 64.6% dos casos. A faixa etária de maior ocorrência da infecção foi de 8 a 15 dias de idade, com 59,6 % dos casos. A partir do resultado desse estudo, pode-se constatar que o protozoário Cryptosporidium spp. encontra-se amplamente distribuído em propriedades do estado de Santa Catarina.

Cryptosporidium spp. is a protozoan that has stood out for being the most common cause of diarrhea and growth retardation in calves. Because it is considered an opportunistic pathogen, it can cause mortality, in general, when associated with other agents that cause enteric pathologies. Cryptosporidiosis occurs by ingesting Cryptosporidium spp oocysts. and the occurrence is associated with poor hygienic conditions, high density of calves in paddocks and the age of the animals. The main objective of this study was to determine the prevalence of infection by Cryptosporidium spp. in calves with the clinical sign of diarrhea, as well as assessing the epidemiological aspects associated with the frequency of infection. The sampling by region was based on the total population, the margin of error and the number of standard deviations, selecting heifers with age ranging from birth to 150 days of age, totaling 425 samples from 141 properties in different regions of the state of Santa Catarina. For diagnosis, the samples were submitted to the Sheather technique followed by modified Ziehl-Neelsen staining. There was a prevalence of 62.1% of positive samples for the protozoan Cryptosporidium spp. in these animals. When analyzing some factors associated with infection by Cryptosporidium spp. in heifers, it can be observed that animals kept in individual facilities presented 69.5% of positive cases, and animals that consumed water from streams were the most affected, with 64.6% of cases. The age group with the highest occurrence of the infection was 8 to 15 days old, with 59.6% of cases. From the result of this study, it can be seen that the protozoan Cryptosporidium spp. it is widely distributed in properties in the state of Santa Catarina.

Detection of mycobacteria in ornamental fish in Iran by culture and Ziehl-Neelsen staining methods

Background: Mycobacterial infections in fish is largely chronic to subacute in nature and affects fishes in fresh water, brackish water and salt water. In addition to their known infectivity to fishes, aquatic mycobacteria pose significant zoonotic concerns. Due to the zoonotic character of the disease, increasing importance of aquariology, and lack of any clinical signs in early steps of mycobacteriosis, present study was undertaken to analyze the distribution of mycobacteria in diseased and apparently healthy ornamental fish from some local aquarium fish shops in four different cities in Iran by culture and Ziehl-Neelsen staining. Materials, Methods & Results: One hundred and one fresh water aquarium fish of 22 species from some local shops in four cities in Iran were examined. Before decontamination, smears of homogenized samples stained with Ziehl-Neelsen. For bacterial culture, samples were inoculated on Lowenstein-Jensen medium. Culture plates were examined daily for four weeks. The rate of growth at different temperature, colony morphology and pigmentation were evaluated for species identification. Among 79 moribund fish examined, 16 individuals were positive for acid fast rods at microscopic examination. Seven fish out of 22 apparently healthy individuals also gave positive microscopic results. Using the culture method, 29 and 10 Mycobacterium isolates were obtained from moribund and healthy fish. The following Mycobacterium species were isolated from unhealthy fish: Mycobacterium fortuitum, M. marinum, and M. smegmatis. The number of different species of Mycobacterium from apparently healthy fish was: M. fortuitum (one fish), M. marinum (one fish), M. terrae (one fish) and M. flavescens (one fish). Discussion: Based on the results in both moribund and apparently healthy fish examined, culture examination showed more mycobacteria than Ziehl-Neelsen staining detection. Lower proportion of Ziehl-Neelsen positive results compared with culture method was reported in aquarium fish in Slovania. Using positive microscopic results, 13 isolates were obtained where as 29 samples gave positive culture results for mycobacteria. Similar result was also observed in clinically healthy ornamental fish and their aquarium environment. The identification of mycobacteria by Ziehl-Neelsen staining is a traditional method. However, acid-fast bacilli may not always be found through direct microscopy because the destruction of mycobacteria or their low number may sometimes happen. Culture examination is a more sensitive method than direct microscopy. However, killing of mycobacteria caused by host defense mechanisms, a low number of viable mycobacteria in the tissue, or by destruction of the mycobacteria during the preparation of the sample could result in negative cultivation results. Species of Mycobacterium identified in unhealthy fish were M. fortuitum, M. marinum and M. smegmatis. High frequency of identifying M. fortuitum and M. marinum (6 out of 7) in the samples provide more evidence that these species are common Mycobacterium species to be found in diseased aquarium fish. Numerous studies showed the common isolation of these two species from aquarium fish. M. marinum infection may be an occupational hazard for certain professionals such as pet shop workers. Many infections may also occur in fish fanciers who keep an aquarium at home. Less common than M. marinum, M. fortuitum is also capable of infecting human. In this study, the occurrence of M. marinum and M. fortuitum in both unhealthy and apparently healthy aquarium fish shows the importance of recognizing fish mycobacteriosis in order to prevent their transmission to human.

Detection of mycobacteria in ornamental fish in Iran by culture and Ziehl-Neelsen staining methods / Detection of mycobacteria in ornamental fish in Iran by culture and Ziehl-Neelsen staining methods

Background: Mycobacterial infections in fish is largely chronic to subacute in nature and affects fishes in fresh water, brackish water and salt water. In addition to their known infectivity to fishes, aquatic mycobacteria pose significant zoonotic concerns. Due to the zoonotic character of the disease, increasing importance of aquariology, and lack of any clinical signs in early steps of mycobacteriosis, present study was undertaken to analyze the distribution of mycobacteria in diseased and apparently healthy ornamental fish from some local aquarium fish shops in four different cities in Iran by culture and Ziehl-Neelsen staining. Materials, Methods & Results: One hundred and one fresh water aquarium fish of 22 species from some local shops in four cities in Iran were examined. Before decontamination, smears of homogenized samples stained with Ziehl-Neelsen. For bacterial culture, samples were inoculated on Lowenstein-Jensen medium. Culture plates were examined daily for four weeks. The rate of growth at different temperature, colony morphology and pigmentation were evaluated for species identification. Among 79 moribund fish examined, 16 individuals were positive for acid fast rods at microscopic examination. Seven fish out of 22 apparently healthy individuals also gave positive microscopic results. Using the culture method, 29 and 10 Mycobacterium isolates were obtained

Background: Mycobacterial infections in fish is largely chronic to subacute in nature and affects fishes in fresh water, brackish water and salt water. In addition to their known infectivity to fishes, aquatic mycobacteria pose significant zoonotic concerns. Due to the zoonotic character of the disease, increasing importance of aquariology, and lack of any clinical signs in early steps of mycobacteriosis, present study was undertaken to analyze the distribution of mycobacteria in diseased and apparently healthy ornamental fish from some local aquarium fish shops in four different cities in Iran by culture and Ziehl-Neelsen staining. Materials, Methods & Results: One hundred and one fresh water aquarium fish of 22 species from some local shops in four cities in Iran were examined. Before decontamination, smears of homogenized samples stained with Ziehl-Neelsen. For bacterial culture, samples were inoculated on Lowenstein-Jensen medium. Culture plates were examined daily for four weeks. The rate of growth at different temperature, colony morphology and pigmentation were evaluated for species identification. Among 79 moribund fish examined, 16 individuals were positive for acid fast rods at microscopic examination. Seven fish out of 22 apparently healthy individuals also gave positive microscopic results. Using the culture method, 29 and 10 Mycobacterium isolates were obtained

Frequência, análise de fatores de risco e comparação de técnicas para detecção de oocistos de Cryptosporidium spp. em fezes de animais de companhia na região Sul do Rio Grande do Sul

Cryptosporidium spp. é um parasito oportunista de caráter zoonótico que infecta o trato gastro intestinal de humanos e outros vários animais tendo a diarreia crônica ou intermitente como principal sintoma da doença. O presente estudo objetivou determinar a frequência de oocistos de Cryptosporidium spp. em fezes de cães e gatos na região sul do Rio Grande do Sul, analisar os fatores de risco que envolvem a criptosporidiose, bem como comparar as técnicas de coloração para o diagnóstico de Cryptosporidium spp. em animais de companhia. Foi disponibilizado um questionário de caráter epidemiológico, abordando perguntas inerentes a possíveis fatores de risco. As variáveis analisadas foram: idade do animal, tipo de alimentação consumida, tipo de água ingerida, moradia, tipo de chão, acesso à rua, interação com outros animais e presença de diarreia. Também foi realizada uma comparação e análise de concordância entre as técnicas de concentração por sedimentação acetato de etila e formalina-éter, e entre os métodos de coloração Kinyoun, Ziehl- Neelsen e safranina para visualização de oocistos de Cryptosporidium spp. em fezes desses animais. Os resultados mostraram que, das 128 amostras (110 cães, 18 gatos), uma frequência verdadeira de 24,6% (25 cães, 2 gatos) foi observada. Quando separadas as espécies de animais, a frequência verdadeira para cães e gatos foi de 26,7% e 12,1%, respectivamente. Nenhuma das quatro amostras de fezes diarreicas obtidas no presente estudo apresentou oocistos. As variáveis que representaram fator de risco foram: comida de preparo, água não tratada e circular exclusivamente em terreno gramíneo. A interação com outros animais, principalmente bovinos, representou 38,5% dos positivos, indicando fator de risco. Na comparação entre as técnicas de coloração, foi possível observar um nível de concordância suficiente a boa entre os dois métodos de concentração por sedimentação como técnica preliminar para coloração de auramina e uma concordância excelente desses mesmos métodos como preliminares para Kinyoun, Ziehl-Neelsen e safranina. A comparação entre os métodos de coloração de Kinyoun e Ziehl-Neelsen para microscopia de luz apresentou fraca concordância, independente do uso de acetato de etila ou formalina-éter. Os métodos de Kinyoun e Ziehl-Neelsen não tiveram qualquer concordância com o método de safranina. Concluíu-se que a frequência de animais de companhia infectados por Cryptosporidium spp. é relevante, destacando que todos os animais positivos do presente estudo não apresentaram diarreia, contribuindo para a contaminação do ambiente e manutenção da infecção. Ademais, pode-se concluir que as técnicas de coloração para microscopia de luz apresentam baixa concordância, indicando diferença de resultados entre um método e outro. O emprego de acetato de etila ou formalina-éter é indiferente para a microscopia ótica.

Cryptosporidium spp. is a facultative parasite that infects the gastrointestinal tract of humans and other animals in which chronic or intermittent diarrhea is the main symptom of the disease. The aim of this study was to investigate the frequency of dogs and cats infected by Cryptosporidium spp. in southern of Rio Grande do Sul using a technique for detecting oocysts in the feces of these animals. In addition, an epidemiological questionnaire with questions considering possible risk factors was made available. We evaluated the following variables: animals age, type of food, water type, home, type of floor, access to the street, interaction with other animals, and presence of diarrhea. We also performed a comparison and concordance analysis between sedimentation concentration techniques (ethyl acetate and formalin ether) and among methods of staining of Cryptosporidium oocysts in the feces. The fenified auramine test was applied to screen the samples because it had excellent specificity and sensitivity. The results showed that of the 128 samples analyzed (110 dogs, 18 cats), a real frequency was 24.6% (25 dogs, cats 2). The true frequency for dogs and cats, separately, was 26.7% and 12.1%, respectively. The four samples of diarrheal feces evaluated in this study did not present oocysts. The variables that represented a risk factor were: Homemade food (p=0.004; OR=3.533, IC.95%=1.442-8.652), non-piped water (0.007; OR=0.291, CI.95%=0.115-0.738) and to transit exclusively on grassy floor (p=0,021, OR=2,778, CI.95%=1.147-6.728). Access to the street showed a protection factor (p = 0.027, OR = 0.242, CI.95% = 0.067-0.868), given that the highest number of parasitized animals consisted of those that did not have access (29.3%). The interaction with other animals, mainly cattle, represented 38.5% of positivity, point to a risk factor (p = 0.010, OR = 3.333, CI.95% = 1.366-8.136). Regarding sedimentation techniques, we observed a satisfactory level of agreement between the two concentration methods, both preliminary to auramine staining (P = 0.000; K = 0.629, CI 95% = 0.457-0.801). The agreement was excellent for the same methods when preliminary to Kinyoun, Ziehl-Neelsen and safranine staining. Comparison between the Kinyoun and Ziehl-Neelsen staining methods to light microscopy showed low agreement regardless of the use of ethyl acetate or formalin ether (p = 0.026, K = 0.391, 95% CI = 0.013-0.769). The methods of Kinyoun and Ziehl-Neelsen did not show agreement with the safranin method. We conclude that the frequency of Cryptosporidium-infected animals is relevant, emphasizing that all the positive animals of this research did not present diarrhea, contributing, consequently, to environmental contamination and maintenance of infection. Moreover, we can conclude that the staining techniques for light microscopy have low correlation, point to difference in results between one and another method. The use of ethyl acetate or formalin-ether is imperceptible to optical microscopy.