Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro Tecnológico, Programa de Pós-Graduação em Engenharia Ambiental, Florianópolis, 2014. / Made available in DSpace on 2015-02-05T20:12:45Z (GMT). No. of bitstreams: 1 327994.pdf: 1803321 bytes, checksum: 56a909b73b9a9f302a055a42abe66ab2 (MD5) Previous issue date: 2014 / Com a crescente preocupação a respeito da poluição hídrica e as possíveis doenças que possam ser causadas através da contaminação com protozoários, surge a filtração em margem como uma das possíveis soluções para o problema, já que pode remover através de filtração os organismos que são resistentes a desinfecção química e algumas vezes podem acabar atingindo as redes de distribuição. Sendo assim, este trabalho objetivou avaliar a eficiência da filtração em margem em colunas de sedimento para remoção de Cryptosporidium spp. através do uso microesferas de poliestireno para simulação deste protozoário. Para consecução desta pesquisa, foi realizado um estudo contendo 5 colunas filtrantes a fim de simular os primeiros metros da filtração em margem, onde foram aplicadas as microesferas simuladoras do protozoário Cryptosporidium spp. Previamente a aplicação das microesferas, procurou-se identificar a quantidade de tempo necessária para que o sistema piloto fosse considerado aclimatado. O controle da qualidade da água produzida pela filtração em margem foi realizado através do monitoramento dos seguintes os parâmetros: carbono orgânico total, coliformes totais, condutividade, cor aparente, cor verdadeira, E. Coli, oxigênio dissolvido, pH, sólidos dissolvidos totais, temperatura e turbidez. A análise das microesferas de poliestireno foi realizada em microscópio de fluorescência. Com base nos resultados obtidos, considera-se aclimatado o sistema piloto após 45 dias de operação, pois nessa altura já é possível alcançar resultados suficientemente bons para se enquadrarem na Portaria nº 2914/2011. Foi também possível concluir que a filtração em margem mostrou-se efetiva na remoção de microesferas de poliestireno a partir de 3 m de leito filtrante. No que diz respeito a turbidez como indicador da presença de Cryptosporidium spp., considera-se que este não corresponde a um indicador confiável, pois houve a presença das microesferas de poliestireno até mesmo quando a turbidez alcançava valores menores que 0,5 uT, como recomenda a legislação brasileira.<br> / Abstract : With the increasing concern about water pollution and possible diseases that may be caused by contamination with protozoa, rises the bank filtration as a possible solution to the problem, since it can remove by filtration, organisms that are resistant to chemical disinfection and can sometimes end up reaching the distribution networks. Thus, this study aimed to evaluate the efficiency of bank filtration by sediment columns for the removal of Cryptosporidium spp. through the use of microspheres for simulation of this protozoan. To achieve this research, was performed a study containing 5 filter columns to simulate the first few meters of bank filtration, where the microspheres simulating the protozoan Cryptosporidium spp. was applied. Previously the application of microspheres, sought to identify the amount of time required for the pilot system to be considered acclimated. The control of the water quality produced by bank filtration was performed by monitoring the following parameters: total organic carbon, total coliform, conductivity, apparent color, true color, E. Coli, dissolved oxygen, pH, total solids dissolved, temperature and turbidity. The analysis of polystyrene microspheres was performed with a fluorescence microscope. Based on the results obtained, it is considered acclimated the pilot system after 45 days of operation, because then it is possible to achieve results good enough to fit the Brazilian legislation. It was also concluded that the bank filtration proved to be effective in the removal of polystyrene microspheres from 3 m filter bed. Regarding the turbidity as an indication of the presence of Cryptosporidium spp., it is considered that this does not correspond to a reliable indicator because there was the presence of polystyrene microspheres even when the turbidity reached values lower than 0,5 NTU, as recommended in Brazilian legislation.

Made available in DSpace on 2012-09-06T01:12:23Z (GMT). No. of bitstreams: 2 license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) 542.pdf: 1674420 bytes, checksum: fbaa2fa72596ec741deb9339aba58df9 (MD5) Previous issue date: 2003 / A qualidade e segurança da água são questões que estão chamando a atenção das autoridades em saúde pública. Várias espécies do gênero Cryptosporidium foram descritas, mas somente o Cryptosporidium parvum têm sido associado às doenças gastrintestinais (...). A criptosporidiose pode ser fatal em imunocomprometidos e pode debilitar severamente indivíduos imunocompetentes. Outro agravante dá-se pelo fato de oocistos de Cryptosporidium serem resistentes às pressões ambientais, podendo sobreviver por vários meses no ambiente aquático e são também resistentes à desinfecção por cloro utilizada no tratamento convencional de água (MULLER, 2000). Atualmente existem diferentes métodos propostos, mais ainda não padronizados. A ausência de uma metodologia padronizada para detecção de oocistos de Cryptosporidium dificulta a viabilização de um monitoramento ou pesquisa deste patógeno emergente. No Brasil, os dados acerca da ocorrência deste parasito em água de abastecimento são limitados. A portaria 1469 de 2000, que em seu padrão microbiológico de poilidade da água para consumo humano recomenda inclusão de pesquisa de organismos patogênicos, com o objetivo de atingir, como meta, um padrão de ausência, dentre outros, oocistos de Cryptosporidium spp. Recomenda-se, mundialmente, o monitoramento de Cryptosporidium em sistema de água potável que abastecem cidades com população entre 10000 e 100000 pessoas. Entretanto, os métodos normalmente usados na determinação de oocisto na água são ineficientes e extremamente variáveis, citando como exemplos a identificação de falsos-positivos por interferências com algas e outras espécies de protozoários (FRANCO et al., 2001). Conclui-se que a inexistência ou inadequação do tratamento de dejetos pode gerar a contaminação dos mananciais de água, podendo veicular e disseminar C. parvum. Em face disso, torna-se fundamental o desenvolvimento e aprimoramento de métodos diagnósticos de detecção.

Tesis para optar al Grado de Magíster en Ciencias Animales y Veterinarias / Cryptosporidium spp. (Familia Cryptosporidiidae), es un grupo de protozoos y parásitos obligados intracelulares que afectan a un amplio rango de vertebrados. En diversos estudios se han descrito a los roedores como hospederos de Cryptosporidium spp., señalando que estos mamíferos presentan la infección de manera natural. Esto los convierte en un riesgo potencial para la salud pública, sobretodo porque muchos roedores contribuyen a la contaminación de agua, suelo y alimentos, al dejar sus deposiciones en todo lugar en donde se desplazan y alimentan. En nuestro conocimiento no existe en Chile ningún registro sobre infección de Cryptosporidium en roedores silvestres. Dado que los cambios en el uso de suelo pueden modificar la estructura y composición de especies, y a su vez, modificar la transmisión de patógenos, en este trabajo se evaluó la presencia y la tasa de infección de Cryptosporidium spp. en heces de roedores silvestres en un área de la zona central de Chile donde existen extensas superficies de plantaciones forestales de pino Monterrey (Pinus radiata). Se analizó si la tasa de infección de Cryptosporidium spp. mostraba variación según distintas variables ambientales (tipo de hábitat, estación del año) y propias de los hospederos (e.g. diversidad de especies, especie de roedor, sexo). Se hipotetizó que las variaciones estructurales en poblaciones y comunidades de roedores silvestres modulan la tasa de infección de Cryptosporidium spp. Se muestrearon roedores silvestres en la Reserva Nacional Los Queules (Región del Maule) y zonas aledañas de plantaciones de pino Monterrey (Pinus radiata) durante el año 2016. Los sitios de muestreo correspondieron a bosque nativo, plantaciones de pino adulta y plantaciones de pino jóvenes. La detección 2 de ooquistes de Cryptosporidium spp. en las heces de los animales se realizó mediante la técnica modificada de tinción de Ziehl-Neelsen. De las 614 muestras analizadas, se detectaron un total de 29 positivas (4,7%) a Cryptosporidium spp., pertenecientes a heces de 4 especies nativas de roedores (5,5% Abrothrix longipilis; 4% Abrothrix olivaceous; 6,6% Oligoryzomys longicaudatus y 3,9% Phyllotis darwini). No se encontraron diferencias en la tasa de infección entre las especies de roedores, tampoco entre los distintos tipos de hábitats ni entre las distintas estaciones del año muestreadas. Se rechazó la hipótesis de esta tesis dado que no hubo evidencia clara en los análisis estadísticos de que la abundancia, riqueza y diversidad de especies puedan modular la tasa de infección de Cryptosporidium spp. Este hallazgo es el primer reporte de Cryptosporidium spp. en roedores silvestres en Chile. La tasa de infección fue comparativamente baja a otros estudios en roedores de otros países, los que pueden llegar a tasas de infección de hasta 68,5%. Las plantaciones de pino no estarían aumentando la tasa de infección de este parásito en roedores, ya que ésta se encontró en porcentajes parecidos en roedores del bosque nativo del área de estudio. Posiblemente otras variables no consideradas en este estudio podrían estar modulando la tasa de infección. Dada la presencia de Cryptosporidium spp. en roedores chilenos, futuros estudios sobre este protozoo en roedores deberían enfocarse en zonas más cercanas a asentamientos humanos para determinar posibles riesgos de infección en la interfaz humano-animales domésticos-animales silvestres / Cryptosporidium spp. (Family Cryptosporidiidae), is a group of protozoa and obligate intracellular parasite, which affect a wide range of vertebrates. Several studies sustain that Cryptosporidium spp. is the most prevalent gastrointestinal parasite in domestic animals and humans. In many studies, rodents have been described as hosts for Cryptosporidium spp., pointing that these mammals present the infection in a natural way. This turns them into a potential risk for public health, mainly because many rodents contribute to water, food and soil contamination as they leave their feces at every place they move around and feed. To our knowledge, there is no record of Cryptosporidium infection in wild rodents in Chile. Due to the fact that land use change can modify species structure and composition and also modify pathogen transmission, this work aimed to evaluate the presence and infection rate of Cryptosporidium spp. in wild rodents from an area of central Chile were vast plantations of Monterrey pine (Pinus radiata) exist. Cryptosporidium spp. infection rate was analyzed from faeces aiming to detect variation according to different environmental variables (habitat type and season) and host variables (e.g. species diversity, rodent species, gender). The hypothesis was that structural variations in rodent populations and communities modulate Cryptosporidium spp. infection rates. Wild rodents from Reserva Nacional Los Queules (Región del Maule) and surrounding areas of Monterrey pine plantations were sampled during 2016. Sampling sites were in native forest, adult pine plantations and young pine plantations. Cryptosporidium spp. oocyst detection in feces was performed by acid-fast method (Ziehl Neelsen). 4 Out of the 614 analyzed samples, 29 were positive to Cryptosporidium spp. (4.7%), belonging to 4 different native rodent species (5,5% Abrothrix longipilis, 4% Abrothrix olivaceous, 6,6% Oligoryzomys longicaudatus y 3,9% Phyllotis darwini). No differences were found on infection rates between rodent species, habitat type, or season. Thus, the hypothesis was rejected, because there was no clear evidence from statistic results that species’ abundance, richness, and diversity can modulate Cryptosporidium spp. infection rates. This finding is the first report of Cryptosporidium spp. in wild rodents in Chile. Infection rate was low compared to other studies in rodents from other different parts of the world, in which it can reach up to 68.5%. Pine plantations are not increasing the infection rates of this parasite in rodents as infection rate in these habitats were similar than the ones from native forest. Presumably other variables not considered in this work could be modulating the infection rate. Given the presence of Cryptosporidium spp. in Chilean wild rodents, future studies about this protozoa in wild rodents should focus in areas closer to human settlements, in order to determine possible risks of infection at the human-domestic animals-wildlife interface / Financiamiento: Proyecto FONDECYT Postdoctorado 3160037

Abstract In order to examine the prevalence of Cryptosporidium in wild Arvicolinae in the Czech Republic and understand the role that wild rodents play in the transmission of this parasite to humans and livestock, 152 faecal samples from 129 common voles (Microtus arvalis) and 23 bank voles (Clethrionomys glareolus) were collected on 9 localities in 2012. All samples were examined for presence of Cryptosporidium sp. using both the aniline-carbol-methyl violet staining method and molecular tools. The age, sex and faecal consistency were noted at the time of sampling. Microscopical examination revealed the presence Cryptosporidium sp. oocysts in 2 samples originated from common voles and 2 samples from bank voles. Genotyping was done through PCR amplification and characterization of the SSU rRNA and actin loci. Cryptosporidium specific DNA was detected in 10 samples (4 from common voles and 6 from bank voles) including those microscopically positive. Cryptosporidium infection was not linked to diarrhoea. Sequence and following phylogeny analyses revealed two new Cryptosporidium genotypes originated from bank voles and two new genotype from common vole, phylogeneticaly distinct from known species and genotypes. The host specificity needs to be verified by experimental infection in the future.

In total 399 samples from three species of squirrels (Sciurus vulgaris, Sciurus carolinensis and Callosciurus finlaysonii) were collected from 2010 to 2013. All samples were examined for the presence of Cryptosporidium using by molecular methods, which demonstrated by presence of Cryptosporidium specific DNA in 18 samples (4.5%). Out of them 12 samples were positive onCryptosporidium ferret genotype, 3 on Cryptosporidium skunk genotype, 1 on Cryptosporidium chipmunk genotype I and 2 samples onCryptosporidium ubiquitum. Statistical analyses did not reveal any influence of gender on the occurrence of Cryptosporidium. Juvenile individuals of squirrels from the family Sciuridae are more often infected by Cryptosporidium than the adult animals.

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Programa de Pós-Graduação em Biotecnologia / Made available in DSpace on 2012-10-20T22:25:47Z (GMT). No. of bitstreams: 0 / O esgoto não tratado e lançado no meio ambiente é um problema que

abstract: The purpose of this study was to determine the applicability of fluorescent microspheres as a surrogate to measure the removal of Cryptosporidium oocysts through the coagulation, flocculation, sedimentation, and filtration steps of conventional water treatment. In order to maintain accuracy and applicability, a local water treatment facility was chosen as the system to model. The city of Chandler Arizona utilizes conventional treatment methodologies to remove pathogens from municipal drinking water and thus the water, coagulant, polymer, and doses concentrations were sourced directly from the plant. Jar testing was performed on four combinations of coagulant, polymer, and fluorescent microsphere to determine if the log removal was similar to that of Cryptosporidium oocysts. Complications with the material properties of the microspheres arose during testing that ultimately yielded unfavorable but conclusive results. Log removal of microspheres did not increase with added coagulant in the predicted manner, though the beads were seen aggregating, the low density of the particles made the sedimentation step inefficient. This result can be explained by the low density of the microspheres as well as the potential presence of residual coagulant present in the system. Given the unfavorable properties of the beads, they do not appear to be a suitable candidate for the surrogacy of Cryptosporidium oocysts in conventional drinking water treatment. The beads in their current state are not an adequate surrogate; however, future testing has been outlined to modify the experiment in such a way that the microspheres should behave like oocysts in terms of physical transportation. / Dissertation/Thesis / Masters Thesis Civil Engineering 2015

Resumo:O objetivo desta pesquisa foi determinar a ocorrência de parasitoses gastrintestinais e caracterizar molecularmente as espécies de Cryptosporidium spp. em amostras fecais de cordeiros de até um ano de idade do município de Alambari, São Paulo. Um total de 193 amostras foram colhidas e em 126 das mesmas foi efetuada a contagem de ovos por gramas de fezes (OPG) Gordon e Whitlock (1939), para determinar a presença de helmintos e coccídeos do gênero Eimeria. Todas as amostras, foram avaliadas por meio da nested-PCR para amplificação de fragmentos de DNA da subunidade 18S do gene do RNAr para Cryptosporidium. Por meio do teste Qui-Quadrado e/ou Fisher foi verificada a associação da presença das parasitoses gastrintestinais em relação ao sexo, raça e idade dos animais. Das 126 amostras analisadas, 71,4% foram positivas pelo método de OPG, com identificação de 58,9% de ovos de Estrongilídeos; 5,6% de oocistos de Eimeria spp.; coinfecções em, 14,4% de Estrongilídeos e Moniezia spp.; e 15,5% Estrongilídeos e Eimeria spp., e 5,6% infecção pelos três parasitos. Coproculturas foram positivas em 16 amostras, com observação dos gêneros: 31,3% Haemonchus spp.; 12,5% Trichostrongylus spp.; Oesophagsotumum spp. em apenas uma; e infecção mista em 50% por Haemonchus e Trichostrongylus. Para Eimeria spp., a ocorrência foi de 19% dos animais, identificando as espécies: E. crandallis; E. faurei; E. marsica; E. ovinoidalis; E. pallida; E. parva e E. weybridgensis. A nPCR revelou positividade de 15% e o sequenciamento foi possível em 19 amostras, com detecção das espécies: C. xiaoi em 15 amostras, C. ubiquituim em três, e C. meleagridis em uma amostra, sendo estas duas últimas consideradas espécies zoonóticas / Abstract:The objective of this research was to determine the occurrence of gastrointestinal parasites and molecularly characterize the species of Cryptosporidium spp. in stool samples from lambs under one year old in the city of Alambari, São Paulo. A total of 193 samples were collected in 126 and the same was done counting eggs per gram of feces (EPG) Gordon and Whitlock (1939), to determine the presence of helminths and coccidia of the genus Eimeria. All samples were evaluated by nested-PCR amplification of DNA fragments subunit 18S rRNA gene for Cryptosporidium. By using the Chi- square and/or Fisher was significant association of the presence of intestinal parasites in relation to gender, race and age of the animals. Of the 126 samples analyzed, 71.4% were positive by OPG, identifying 58.9% of eggs of Strongyles, 5.6% of oocysts of Eimeria spp.; Coinfection in 14.4% of Strongyles and Moniezia spp., and 15.5% Strongyles and Eimeria spp., and 5.6% infection by the three parasites. Stool cultures were positive in 16 samples, with observation of genres: 31.3% Haemonchus spp., 12.5% Trichostrongylus spp.; Oesophagsotumum spp. in one, and 50% mixed infection with Haemonchus and Trichostrongylus. To Eimeria spp., the incidence was 19% of the animals, identifying the species: E. crandallis; E. faurei; E. marsica; E. ovinoidalis; E. pallida; E. parva and E. weybridgensis. The nPCR was positive in 15% and sequencing was performed in 19 samples with detection of species: C. xiaoi in 15 samples, C. ubiquituim three, and C. meleagridis in a sample, the latter being considered zoonotic species / Orientador:Katia denise Saraiva Bresciani / Banca:Jancarlo Ferreira Gomes / Banca:Suely Regina Mogami Bomfim / Mestre

Made available in DSpace on 2014-06-11T19:27:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-04-26Bitstream added on 2014-06-13T20:56:05Z : No. of bitstreams: 1 000737488.pdf: 765065 bytes, checksum: 2838b9b15f0909a0ee3d2a23a324a1dd (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / The objective of this research was to determine the occurrence of gastrointestinal parasites and molecularly characterize the species of Cryptosporidium spp. in stool samples from lambs under one year old in the city of Alambari, São Paulo. A total of 193 samples were collected in 126 and the same was done counting eggs per gram of feces (EPG) Gordon and Whitlock (1939), to determine the presence of helminths and coccidia of the genus Eimeria. All samples were evaluated by nested-PCR amplification of DNA fragments subunit 18S rRNA gene for Cryptosporidium. By using the Chi- square and/or Fisher was significant association of the presence of intestinal parasites in relation to gender, race and age of the animals. Of the 126 samples analyzed, 71.4% were positive by OPG, identifying 58.9% of eggs of Strongyles, 5.6% of oocysts of Eimeria spp.; Coinfection in 14.4% of Strongyles and Moniezia spp., and 15.5% Strongyles and Eimeria spp., and 5.6% infection by the three parasites. Stool cultures were positive in 16 samples, with observation of genres: 31.3% Haemonchus spp., 12.5% Trichostrongylus spp.; Oesophagsotumum spp. in one, and 50% mixed infection with Haemonchus and Trichostrongylus. To Eimeria spp., the incidence was 19% of the animals, identifying the species: E. crandallis; E. faurei; E. marsica; E. ovinoidalis; E. pallida; E. parva and E. weybridgensis. The nPCR was positive in 15% and sequencing was performed in 19 samples with detection of species: C. xiaoi in 15 samples, C. ubiquituim three, and C. meleagridis in a sample, the latter being considered zoonotic species / FAPESP: 10/52542-3

Orientador: Marcelo Vasconcelos Meireles / Banca: Katia Denise Saraiva Bresciani / Banca: Raphael Lucio Andreatti Filho / Resumo: Devido à escassez de informações referentes à ocorrência de Cryptosporidium em aves selvagens no Brasil, este projeto foi realizado visando à detecção de Cryptosporidium em amostras de fezes de várias espécies de aves selvagens da fauna brasileira, por meio da reação em cadeia de polimerase - "nested" (n-PCR). Um total de 488 amostras de 146 espécies de aves selvagens foi coletado em zoológicos, criatórios, Hospitais Veterinários e residências, para extração do DNA genômico de oocistos e realização de n-PCR, visando à amplificação de fragmentos dos genes da subunidade 18S do RNA ribossômico e da actina. Foi observada amplificação para Cryptosporidium em 20 (4,7%) das amostras examinadas. Por meio de sequenciamento dos fragmentos de DNA amplificados foram identificadas duas espécies de Cryptosporidium: C. galli em curiós (Oryzoborus angolensis) e C. baileyi em urubu-de-cabeça-preta (Coragyps atratus). / Abstract: Due to the paucity of data related to the occurrence of Cryptosporidium in wild birds from Brazilian fauna, this research was accomplished in order to screen fecal samples from various species of wild birds in Brazil for the presence of Cryptosporidium by the use of nested polymerase chain reaction (n-PCR). A total of 488 fecal samples from 146 species of wild birds were collected in Zoos, Veterinary Hospitals, breeder's facilities and residences. After extraction of genomic DNA from oocysts n-PCR was accomplished for amplification of fragments from 18S subunit of the ribosomal RNA gene and from actin gene. Positive amplification for Cryptosporidium was obtained in 20 (4.09%) samples. Sequencing of amplified fragments allowed the identification of C. galli in lesser-seed finches (Oryzoborus angolensis) and C. baileyi in black vulture (Coragyps atratus). / Mestre