Made available in DSpace on 2016-04-28T20:16:19Z (GMT). No. of bitstreams: 1 Bruno Pereira Berto1.pdf: 2100000 bytes, checksum: 7c975b30da67285bf2c8d87c9e105bc9 (MD5) Previous issue date: 2010-03-02 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Coccidiosis associated with the genera Eimeria Schneider, 1875 and Isospora Schneider, 1881 in the order Passeriformes are reported for more than two centuries. This study aimed to contribute to the morphology and systematic of coccidian parasites of the order Passeriformes, providing scientific basis for identification of parasite species of birds from North, South and Central America. The coccidia were organized and grouped according to the family of the host, following the concept widely recognized of family-specificity and the systematic of the class Aves updated. Isospora tiesangui, I. marambaiensis, I. sepetibensis, I. cadimi, I. navarroi, I. ramphoceli, I. mionectesi, I. feroxis, I. cagasebi, I. coerebae, I. piacobrai e Eimeria sicki were identified and characterized according to their respective hosts of the order Passeriformes, which inhabit the Atlantic forest of the Marambaia Island, Rio de Janeiro, Brazil. The main feature of differentiation and identification of these species was the Stieda and substieda bodies, since the morphometric parameters did not provide sufficient differentiation. The specificity of coccidia occurred at the family level, because Ramphocelus bresilius dorsalis and the new hosts Dacnis cayana and Thraupis palmarum, family Thraupidae, were described for the species I. tiesangui, I sepetibensis and I. navarroi, and, similarly, Myiarchus ferox and Leptopogon amaurocephalus, family Tyrannidae, were described for E. sicki. Finally, dichotomous keys for identification were effective for the families Thraupidae and Tyrannidae. / Coccidioses associadas aos g?neros Eimeria e Isospora na ordem Passeriformes s?o relatadas h? mais de dois s?culos. Este trabalho objetivou contribuir para a morfologia e sistem?tica de cocc?dios parasitos da ordem Passeriformes, fornecendo embasamento cient?fico para identifica??o de esp?cies parasitas de aves das Am?ricas do Norte, do Sul e Central. Os cocc?dios foram organizados e agrupados de acordo com a fam?lia do hospedeiro, seguindo o conceito fam?lia-espec?fico, amplamente reconhecido e a sistem?tica da classe Aves atualizada. Isospora tiesangui, I. marambaiensis, I. sepetibensis, I. cadimi, I. navarroi, I. ramphoceli, I. mionectesi, I. feroxis, I. cagasebi, I. coerebae, I. piacobrai e Eimeria sicki foram identificadas e caracterizadas de acordo com seus respectivos hospedeiros da ordem Passeriformes, os quais habitam o bi?topo de sub-bosque da Mata Atl?ntica, Ilha da Marambaia, Rio de Janeiro, Brasil. A principal caracter?stica de diferencia??o e identifica??o destas esp?cies foi o complexo corpo de Stieda e substieda, uma vez que o estudo morfom?trico n?o forneceu par?metros suficientes de diferencia??o. A especificidade ocorreu em n?vel de fam?lia, pelo fato de Ramphocelus bresilius dorsalis e os novos hospedeiros Dacnis cayana e Thraupis palmarum, da fam?lia Thraupidae, terem sido descritos para as esp?cies I. tiesangui, I. sepetibensis e I. navarroi, e, da mesma forma, Myiarchus ferox e Leptopogon amaurocephalus, da fam?lia Tyrannidae, foram descritos para E. sicki. Por fim, chaves dicot?micas de identifica??o de esp?cies de cocc?dios parasitas de aves Passeriformes foram efetivadas para as fam?lias Thraupidae e Tyrannidae.

Pooled faecal specimens of pigs were collected from the floor on two farms (A and B) and parasitologically examined. Investigations were divided into the five period: winter 2005 (A), spring 2005 (A), autumn 2005 (B), winter 2006 (B) and spring 2006 (B). In total, we collected 249 faecal specimens of pigs, of them 91 specimens were collected from farm A and 158 specimens from farm B. In farm A we collected 32 specimens from post-weaned piglets and in farm B 20 specimens from post-weaned piglets. Piglets were housed on litter stabling on both farms.

In den vorliegenden Studien wurden verschiedene diagnostische Verfahren zum Nachweis von Protozoen und Nematoden im Hinblick auf Sensitivität, Arbeitsaufwand und Kosten miteinander verglichen. Zudem wurde die Eignung der PCR zur molekularen Charakterisierung der Cryptosporidium spp. exemplarisch an Igelkotproben getestet. Bei der Untersuchung von 90 Ferkelkotproben auf I. suis war die Sensitivität eines Kotausstriches mit nachfolgender Autofluoreszenzmikroskopie (AM) signifikant höher als bei einem Flotationsverfahren (FV) mit NaCl-Zucker-Lösung und bei dem kombinierten Sedimentations-Flotations-Verfahren (KSFV) mit verschiedenen Flotationslösungen (NaCl, ZnSO4, NaCl-Zucker-Lösung) mit nachfolgender Lichtmikroskopie. Zudem ist der Arbeitsaufwand für die AM deutlich geringer als bei dem FV und KSFV. Die höheren apparativen Kosten für die AM sind bei hohem Probendurchsatz durch den geringeren Zeitaufwand und der höheren Sensitivität gerechtfertigt. Die Anzahl Kryptosporidien-positiver Proben war bei der Untersuchung von 103 Kälberkotproben auf Cryptosporidium sp. mittels Enzymimmunoassays (EIA; ProSpecT® Cryptosporidium Microplate Assay) im Vergleich zur Karbolfuchsin-Färbung (CF) nach HEINE (1981) und der modifizierten-Ziehl-Neelsen-Färbung (MZN) nach HENRIKSEN u. POHLENZ (1982) am höchsten und signifikant höher als bei der Anwendung der MZN, wenn 10 Blickfelder durchmustert wurden. Bei der Untersuchung von 74 Igelkotproben auf Cryptosporidium sp. mittels EIA (ProSpecT®), einem immunochromatographischen Verfahren (FASTest® CRYPTO Strip), der MZN nach HENRIKSEN u. POHLENZ (1981) und einem direkten Immunfluoreszenz-Test (IFA; MERIFLUOR Cryptosporidium/Giardia) wurden in 9 (EIA), 10 (FASTest®), 11 (MZN) und 12 (IFA) Proben Cryptosporidium sp. nachgewiesen. Der Arbeitsaufwand des FASTest® und der CF ist mit dem EIA vergleichbar, während der IFA und die MZN mehr Zeit benötigen. Die Anwendung des FASTest®, des IFA und des EIA ist mit höheren Kosten verbunden als bei den Färbemethoden, können aber gut in den Arbeitsablauf eines diagnostischen Labors eingefügt werden und sind einfach auszuwerten. Darüber hinaus wurden 45 Kotproben, welche bis zu 27 Tage bei verschiedenen Temperaturen (+6 °C, +16 °C, +30 °C, +40 °C) gelagert wurden, untersucht, um einen Einfluss der Temperatur auf das Untersuchungsergebnis von EIA, CF und MZN zu ermitteln. Während sich die Anzahl positiver Proben bei der Untersuchung mit den Färbemethoden temperatur- und zeitabhängig reduzierte, wurde das Untersuchungsergebnis mittels EIA von der Lagerungstemperatur nicht beeinflusst, so dass ungekühlt transportierte Proben vorzugsweise mit dem EIA untersucht werden sollten. Dagegen ist die CF aufgrund ihrer einfachen und preiswerten Durchführung zur Untersuchung einer hohen Anzahl an Proben geeignet, sofern eine ununterbrochene Kühlung der Proben gewährleistet ist und diese innerhalb von drei Tagen untersucht werden. Der FASTest® ist zur Anwendung in Tierarztpraxen und Ställen geeignet, da zur Untersuchung kein Mikroskop benötigt wird und die Resultate schnell vorliegen. Die Verwendung des IFA, der Kryptosporidien-Oozysten und Giardien-Zysten nachweist, bietet sich vor allem bei Proben an, die auf beide Protozoen untersucht werden sollen. Das Vorkommen der Kryptosporidiose bei unterentwickelten und geschwächten Igeln, welche zum Überwintern in Igelstationen aufgenommen werden, ist hoch. Von 188 untersuchten Igelkotproben konnten in 29,8 % der Proben Cryptosporidium spp. nachgewiesen werden. Durch die Genotypisierung der Kryptosporidien aus 15 positiven Igelkotproben mittels RFLP-PCR basierend auf dem 18S rRNA-Gen konnte in allen untersuchten Proben die Präsenz von C. parvum gezeigt werden. Mit Hilfe der Multilocus-Sequenz-Typisierung der Fragmente des 60kDa Glycoprotein-Gens, des 18S rRNA-Gens, des Actin-Gens und des 70 kDa Hitzeschockprotein-Gens konnten drei verschiedene Subtypen-Familien (IIa, IIc und eine neue als VIIa vorgeschlagene Subtypen-Familie) erkannt werden. Die von den Igeln ausgeschiedenen Kryptosporidien-Oozysten mit zum Teil nachgewiesenem zoonotischen Potential (IIa Subtypen-Familie) könnten eine Infektionsquelle für den Menschen sein, aber auch ein antropozoonotisches Potential (IIc Subtypen-Familie) sollte in Betracht gezogen werden, so dass die Hygiene in den Igelstationen einen hohen Stellenwert einnehmen sollte. Die Untersuchungsergebnisse zum Nachweis von Eimeria-Arten beim Kalb von 70 Sammelkotproben, hergestellt aus 10 Einzelkotproben (SKP10), bzw. von 30 Sammelkotproben, zusammengesetzt aus 5 Einzelkotproben (SKP5), wurden mit denen der zugehörigen Einzelkotproben (EKP) verglichen. Die Resultate der EKP (arithmetischer Mittelwert) und der zugehörigen SKP weisen mit den signifikant häufigeren Abweichungen im Bereich von bis zu 100 Oozysten pro Gramm Kot (OpG) eine geringe Differenz zwischen den beiden Verfahren auf. Durch den sicheren Nachweis von Eimeria-Oozysten bei einem erwarteten Oozystengehalt von nur 202 OpG (SKP10) und 122 OpG (SKP5) ist die Untersuchung von Kälbersammelkotproben, eine Methode mit geringem Arbeitsaufwand und geringen Untersuchungskosten, zum Nachweis einer klinischen oder subklinischen Kokzidiose geeignet. Bei 51 Pferdekotproben wurde jeweils dreimal das kombinierte Sedimentations-Flotations-Verfahren (KSFV), wobei die Entnahme von verschiedenen Lokalisationen der Kotprobe (aus der Randregion, dem Inneren oder aus beiden Lokalisationen) erfolgte, und jeweils dreimal das KSFV mit vorheriger Homogenisierung einer größeren Kotmenge zum Nachweis von Nematodeneier durchgeführt. Eine Anhäufung der Strongyliden- und Ascarideneier in einem bestimmten Bereich der Proben konnte durch die Untersuchungen der verschiedenen Lokalisationen (á 10 g Kot) nicht nachgewiesen werden, so dass eine weitgehend homogene Verteilung dieser Nematodeneier in einer Pferdekotprobe wahrscheinlich ist. Zudem konnten die Untersuchungsergebnisse des KSFV, bei welchem 10 g Kot untersucht werden, durch die vorherige Homogenisierung einer größeren Probenmenge nicht verbessert werden. Zum Nachweis von Nematoden beim Pferd sollte dem Labor eine ausreichende Probenmenge (ca. 50 g) zugesandt werden. Die Homogenisierung einer größeren Probenmenge vor der Durchführung einer diagnostischen Methode, bei der Aliquote von mindestens 10 g Kot Verwendung finden, ist unnötig. / The studies presented were carried out to compare different diagnostic methods for detection of protozoa and nematodes regarding sensitivity, expenditure of human labour and costs. Besides, the ability of the PCR for the molecular characterization of the Cryptosporidium spp. was tested exemplarily in faecal samples of hegdehogs. The examination of ninety faecal samples of suckling piglets showed a significantly higher sensitivity of faecal smears examined by autofluorescence microscopy (AM) compared to the flotation method (FV) using NaCl-sucrose solution and the combined sedimentation-flotation method (KSFV) using different flotation solutions (NaCl, ZnSO4, NaCl-sucrose) scanned by bright field microscopy. Moreover the expenditure of human labour by AM is considerably lower than FV and KSFV. The costs related to equipment for AM is justified in case of high sample throughput and by superior sensitivity. The enzyme immunoassay (EIA; ProSpecT® Cryptosporidium Microplate Assay) was the most sensitive method for diagnosis of cryptosporidiosis in calves (n = 103) compared to the carbol fuchsin (CF; HEINE 1981) and modified Ziehl-Neelsen (MZN; HENRIKSEN a. POHLENZ 1982) staining techniques. The sensitivity of the EIA was significantly higher than the MZN, if ten fields of view were scanned. 74 faecal samples of hedgehogs were examined with the EIA (ProSpecT®), an immunochromatographic method (FASTest® CRYPTO Strip), the MZN (HENRIKSEN u. POHLENZ (1981)) and a direct immunofluorescent assay (IFA; MERIFLUOR Cryptosporidium/Giardia). Cryptosporidium sp. were detected in 9 (EIA), 10 (FASTest®), 11 (MZN) und 12 (IFA) faecal samples. The hands on time of the FASTest® and CF is comparable to EIA while the IFA and MZN are more time-consuming. The examination of the FASTest®, IFA and EIA is combined with higher costs than the staining techniques, but they can be integrated in the work flow of a routine diagnostic laboratory easily and evaluation is simple. Moreover 45 faecal samples stored up to 27 days at different temperature (+6 °C, +16 °C, +30 °C, +40 °C) were examined to evaluate the influence of temperature on the results of EIA, CF and MZN. While the number of the positive samples of stained smears decreased in a temperature and time-dependent manner, the results of the EIA were not influenced by sample storage at any temperature, so that samples transported without cooling should be examined preferably by EIA. Nevertheless the CF due to its simplicity and low costs is suited for scanning of a high number of samples, if they were cooled continuously and examined within three days. The FASTest® is qualified for use in veterinary practice and stables, because the examination requires no microscope and the results are obtained immediately. The IFA, which can detect Crypotsporidium oocysts as well as Giardia cysts, is suited especially for faecal samples suspected to contain both protozoa. Cryptosporidial infections are very frequent in hedgehogs which are admitted for hibernation to hedgehog rehabilitation centres because of their insufficient body weight and weakness. Cryptosporidium spp. were detected in 29.8 % of 188 faecal samples of hedgehogs. The genotyping of Cryptosporidium spp. by PCR and RFLP-PCR based on the 18S ribosomal RNA gene were performed on 15 faecal samples of hedgehogs positive for Cryptosporidium spp. and suggested the presence of C. parvum in all samples. Multilocus sequence typing on partial 60 kDa glycoprotein gene, 18S rRNA gene, actine gene, 70 kDa heat shock protein gene sequences revealed 3 different subtype families: IIa, IIc and a new proposed as VIIa subtype family. Some of the Cryptosporidium oocysts excreted from hedgehogs are zoonotical (IIa subtype family) or anthropozoonotic(IIc subtype family). Thus hygienic measurements to avoid transmission are essential in hedgehog rehabilitation centres. The results of examination of 70 pooled faecal samples originating from 10 calves (SKP10) and 30 pooled faecal samples originating from 5 calves (SKP5) for detection of Eimeria spp. were compared with the arithmetic means of opg (oocysts per gram of faeces) counts of the respective single 10 or 5 samples. A low difference between both methods of less than 100 opg was significantly more frequently observed than higher differences. Low values of 202 opg and 122 opg were reliably detected in SKP10 und SKP5, respectively, and thus examination of pooled faecal samples appears to be suitably sensitive and cost effective to detect clinical and subclinical coccidiosis in calves. 51 faecal samples of horses were examined three times by KSFV for nematode eggs by taking aliquots from different locations of the same faecal samples (from the margin, from inside and from both locations). Thereafter the KSFV with the homogenisation of a larger amount of faeces was also carried out three times. The examination of samples from the different locations (each 10 g of faeces) delivered no evidence for accumulation of nematode eggs (strongyles and Parascaris equorum) in the faeces and thus the distribution of the nematode eggs appears sufficiently homogeneous in faecal samples of horses. Homogenisation of a larger amount of faeces did not improve the results of coproscopy. For diagnostic purposes 50 g faeces per sample should be shipped to the laboratory. The homogenisation of a larger amount of faeces before using a diagnostic method is dispensable, if aliquots of 10 g faeces are examined.

Made available in DSpace on 2014-07-29T15:29:13Z (GMT). No. of bitstreams: 1 Dissertacao Solimar A de Oliveira.pdf: 1870870 bytes, checksum: 5c7849377c8610cb5cbc8b8d4905a909 (MD5) Previous issue date: 2012-02-10 / Introduction: The chronic renal insufficiency is in between the transition epidemiologist illness, being able to be affected by the enteric opportunistic parasites for representing a population of immunosuppressed. Catalogued as emergent agents of opportunistic character, protozoan disease is responsible for important morbi-mortality rates, but little recognized on the part of the professionals of health and for the shortage of specialized laboratories in its diagnostics. They are caused mainly by protozoan, as the Blastocystis hominis, Cryptosporidium sp, Isospora belli, Cyclospora cayetanensis, amongst others. Objectives: Mapping world-wide studies through a systematic revision of literature concerned to the detection of these protozoan in hemodialysis patients. And, besides, to identify enteric opportunistic agents in immunosuppressed children with chronic nephropathies who were submitted to hemodialysis and also children patients who don t have chronic nephropathies, in the Clinical Hospital /UFG. Methods: The theoretical part, represented by the systematic revision of literature, was elaborated from standardized forms on the selection of scientific articles available in the Virtual Library in Health. This work, concerning the experimental part, was built in the period of October of 2009 to May of 2011 with analysis of the epidemiologic profiles of the patients and laboratorial detection of opportunistic enteroparasitosis in 229 fecal samples of 26 hemodialysis children (test-group) and of 59 children who haven t chronic nephropathies (control group), from the Hospital of the Clinics of the Federal University of Goiás. For further detection of the oocysts of coccids (Cryptosporidium sp, Cyclospora cayetanensis and Isospora belli), microscopic examination was used direct (fresh).It was also used the methods of Hoffman, Pons and Janer, Ridley or of concentration in formalin 10% - Acetate of Ethyl, Coloration of Kinyoun (hot), and, Ziehl-Neelsen (modified). With regard to the diagnosis of Blastocystis hominis, it was used the microscopic examination direct (fresh) and the technique of Coloration of Nair - Blue of Methylene. Results: In the systematic revision of literature, nine articles had been selected, and from the interpretation on these studies, the presence of enteric opportunistic protozoan in fecal samples of hemodialysis patients was evidenced. In the experimental part, the detection of protozoan for patients, in the test group and in the control group was, respectively, of: Blastocystis hominis in 9 (34,6%) and 13 (22%); Giardia lamblia in 3 (11,5%) and 2 (3,4%); Endolimax nana in 9 (34,6%) and 9 (15,3%); Entamoeba coli in 3 (11,5%) and 2 (3,4%). And still it had been detected only in the test group: Cryptosporidium sp in 1 (3,8%) patient and Entamoeba histolytica/dispar in 3 (11,5%). Regarding the quantitative analysis of fecal samples, it was collected 115 samples of the group of hemodialysis children and 114 samples of the group of children who don t have chronic nephropathies. The results gotten in this comparison had designated the presence of oocysts of intestinal protozoan in the test group and in the control group. Respectively, we have: Blastocystis hominis in 24 (20,87%) and 16 (14,04%) samples; Giardia lamblia in 3 (2,61%) and 2 (1,75%) samples; Endolimax nana 15 (13,4%) and 9 (7,89%) samples. Besides, it had been detected only in the test group: Cryptosporidium sp in 1 (0,87%) sample and Entamoeba histolytica/dispar in 3 (2,61%). With regard to the diarrheal feces analysis, it was detected in test group 1 sample (0,87%), and in the control group, 8 (7.02%). Conclusion: These findings demonstrate that such agents are present in our environment, reinforcing isolated infections or associates, between them or ahead of other opportunistic enteric parasites, providing a risk for the population of hemodialysis patient. They still disclose the urgency of an implantation of specialized laboratories with specific detection techniques of these infectum-parasitic agents. / Introdução: A insuficiência renal crônica está entre as doenças de transição epidemiológica, podendo ser afetada pelas parasitoses entérico oportunistas por representar uma população de imunossuprimidos. Catalogadas como agentes emergentes de caráter oportunista, as protozooses são responsáveis por importantes índices de morbi-mortalidade, mas pouco reconhecidas por parte dos profissionais de saúde e pela escassez de laboratórios especializados em seus diagnósticos. São causadas, principalmente, por protozoários como o Blastocystis hominis, Cryptosporidium sp, Isospora belli, Cyclospora cayetanensis, dentre outros. Objetivos: Mapear estudos mundiais mediante revisão sistemática da literatura quanto à detecção destes protozoários em pacientes hemodialisados. Identificar agentes entéricos oportunistas em crianças imunossuprimidas com nefropatias crônicas e submetidas à hemodiálise e em crianças não portadoras de nefropatias crônicas, no Hospital das Clínicas/UFG. Métodos: A parte teórica, representada pela revisão sistemática da literatura, foi elaborada a partir de formulários padronizados para a seleção de artigos científicos existentes na Biblioteca Virtual em Saúde. Este trabalho, no que tange à parte experimental, foi realizado no período de outubro de 2009 a maio de 2011, com análise do perfil epidemiológico dos pacientes e detecção laboratorial de enteroparasitoses oportunistas em 229 amostras fecais de 26 crianças hemodialisadas (grupo teste) e de 59 crianças não portadoras de nefropatias crônicas (grupo controle), procedentes do Hospital das Clínicas da Universidade Federal de Goiás. Para a detecção dos oocistos de coccídeos (Cryptosporidium sp, Cyclospora cayetanensis e Isospora belli), foram utilizados exames microscópicos diretos a fresco, bem como os métodos de Hoffman, Pons e Janer, Ridley ou de concentração em formalina a 10% Acetato de Etila, Coloração de Kinyoun a quente e ainda, Ziehl-Neelsen modificado. Já para o diagnóstico de Blastocystis hominis, foram utilizados exames microscópicos diretos a fresco e a técnica de Coloração de Nair Azul de Metileno. Resultados: Na revisão sistemática da literatura foram selecionados nove artigos e a partir da interpretação desses estudos foi constatada a presença de protozoários entéricos oportunistas em amostras fecais de pacientes hemodialisados. Na parte experimental, a detecção de protozoários por pacientes, no grupo teste e no grupo controle, foi de: Blastocystis hominis em 9 (34,6%) e 13 (22%); Giardia lamblia em 3 (11,5%) e 2 (3,4%); Endolimax nana em 9 (34,6%) e 9 (15,3%); Entamoeba coli em 3 (11,5%) e 2 (3,4%), respectivamente. Ainda, foram detectados no grupo teste: Cryptosporidium sp em 1 (3,8%) paciente e Entamoeba histolytica/dispar em 3 (11,5%). Quanto à análise quantitativa de amostras fecais, foram coletadas 115 amostras fecais do grupo de crianças hemodialisadas e 114 amostras fecais do grupo de crianças não portadoras de nefropatias crônicas. Os resultados obtidos nessa comparação assinalaram a presença de cistos e oocistos de protozoários intestinais no grupo teste e no grupo controle. Nos referidos grupos teste e controle foram encontrados cistos de Blastocystis hominis em 24 (20,87%) e 16 (14,04%) amostras; Giardia lamblia em 3 (2,61%) e 2 (1,75%) amostras; Endolimax nana 15 (13,4%) e 9 (7,89%) amostras, respectivamente. Além disso, foram detectados no grupo-teste: Cryptosporidium sp em 1 (0,87%) amostra e Entamoeba histolytica/dispar em 3 (2,61%). Em relação à consistência das fezes, foi detectada fezes diarréicas em 1 (0,87%) amostra do grupo-teste e 8 (7,02%) do grupo controle. Conclusão: Estes achados demonstram que tais agentes estão presentes em nosso meio ambiente, potencializando infecções isoladas ou associadas, entre eles ou diante de outros parasitos entéricos oportunistas, proporcionando um risco para a população de hemodialisados. Revelam ainda, a premência de implantação de laboratórios especializados com técnicas específicas de detecção destes agentes infecto-parasitários.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2016-10-20T10:49:41Z No. of bitstreams: 1 2016 - Mariana Borges Rodrigues.pdf: 10445208 bytes, checksum: 3b8788e9719ae12da12f01e329a82878 (MD5) / Made available in DSpace on 2016-10-20T10:49:41Z (GMT). No. of bitstreams: 1 2016 - Mariana Borges Rodrigues.pdf: 10445208 bytes, checksum: 3b8788e9719ae12da12f01e329a82878 (MD5) Previous issue date: 2016-02-19 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Coccidia are obligate intracellular parasites, classified in Subphylum Apicomplexa and order Eucoccidiorida, which has different stages in their life cycles. In Passeriformes, the coccidian species of Isospora Schneider, 1881 are the most common, being the family Thraupidae one of the major families, with 12 host species described. The aim of this study was to identify, characterize and quantify the coccidian parasites of Thraupidae in the Itatiaia National Park. Isospora ramphoceli Berto, Flausino, Luz, Ferreira, Lopes, 2010 was identified in ruby-crowned tanagers Tachyphonus coronatus (Vieillot, 1822), which became a new host, and the Itatiaia National Park a new location for this coccidian species. The intensity of infection in different hosts were high, which can be justified by frugivorous feeding habits that favoring the feco-oral transmission of coccidia and by the positive hosts inhabit in disturbed areas susciptible to the effects of forest edge. The oocysts were characterized as uniform in T. coronatus and morphologically and morphometrically similar to the original description in Ramphocelus bresilius dorsalis Sclater, 1855 on the island of Marambaia, RJ. The specificity of I. ramphoceli occurred at the family level, because T. coronatus and R. b. dorsalis are included in the Thraupidae family. / Os cocc?dios s?o parasitas intracelulares obrigat?rios, classificados no Subfilo Apicomplexa e ordem Eucoccidiorida, que tem fases diferentes em seus ciclos de vida. Em Passeriformes as esp?cies de Isospora Schneider, 1881 s?o as mais comuns, sendo a fam?lia Thraupidae uma das principais fam?lias-hospedeiras com 12 esp?cies descritas. O objetivo deste estudo foi identificar, caracterizar e quantificar os cocc?dios parasitos de Thraupidae do Parque Nacional do Itatiaia. Isospora ramphoceli Berto, Flausino, Luz, Ferreira, Lopes, 2010 foi identificada em ti?s-pretos Tachyphonus coronatus (Vieillot, 1822), o qual se tornou novo hospedeiro e o Parque Nacional do Itatiaia nova localidade para este cocc?dio. As intensidades de infec??o em diferentes hospedeiros positivos foram altas, o que pode ser justificado pelo h?bito alimentar frug?voro que favorece a transmiss?o feco-oral dos cocc?dios e por parte dos hospedeiros positivos habitarem em ?reas antropizadas submetidas aos efeitos de borda de mata. Os oocistos foram caracterizados como uniformes em T. coronatus e morfologicamente e morfometricamente semelhantes a descri??o original em Ramphocelus bresilius dorsalis Sclater, 1855 na Ilha da Marambaia, RJ. A especificidade de I. ramphoceli ocorreu em n?vel de fam?lia, pelo fato de T. coronatus e R. b. dorsalis estarem classificados entre os traup?deos.

Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2017-05-05T14:21:03Z No. of bitstreams: 1 2012 - Cleide Domingues Coelho.pdf: 19229976 bytes, checksum: 94c069047adceba0bbb1974c338db08e (MD5) / Made available in DSpace on 2017-05-05T14:21:03Z (GMT). No. of bitstreams: 1 2012 - Cleide Domingues Coelho.pdf: 19229976 bytes, checksum: 94c069047adceba0bbb1974c338db08e (MD5) Previous issue date: 2012-02-28 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Parasites can affect the physical condition, survival and reproduction of birds may be an important factor in the life history of the host, exerting strong pressure ecological and evolutionary. Among the most important parasites affecting passerine birds, Isospora species were included, and the oocysts counts were used for estimating infection in wild birds as well as essential for studies of prevalence, diagnosis and control of coccidiosis in birds from seizures of wild animals, and keeping under quarantine at Wild Animal Sorting Center and latter for the release. This study aimed to determine the circadian rhythm or periodicity in the elimination of oocysts of Isospora species in Passeriformes, and identify the species of parasite found and verify the effectiveness and prophylaxis of anticoccidial drugs during the quarantine period. In a total of 1393 fecal samples were collected from birds of the order Passeriformes belonging to different families and species, from the apprehensions of wild animals and sent to CETAS (Wild Animal Sorting Center)/IBAMA at Municipality of Serop?dica in the State of Rio de Janeiro. After a period of sporulation, the samples were subjected to centrifugal flotation technique with sucrose, quantified and the results expressed in OoPD (oocysts per defecation). The results showed that, regardless of the continent where the birds live, photoperiod is an important factor in maintaining the schedule for the elimination of oocysts of the genus Isospora. Birds of several families had an OoPD means, in relationship of shedding oocysts in the feces, the highest eliminations is in the afternoon. For control of coccidiosis in these birds, throughout the use of anticoccidial drugs were observed that the effectiveness may vary with the species of the parasite and the birds, because they have different feeding habits and behavior, which may influence the response to treatment / Os parasitos podem afetar a condi??o f?sica, sobreviv?ncia e reprodu??o das aves, podendo ser um importante fator na hist?ria de vida do hospedeiro, exercendo forte press?o ecol?gica e evolucion?ria. Dentre os parasitos mais importantes que afetam os Passeriformes podemos citar os cocc?dios do g?nero Isospora, e a estimativa da infec??o em p?ssaros silvestres ? essencial para os estudos de preval?ncia, diagn?stico e controle deste cocc?dio nas aves oriundas de apreens?es do tr?fico de animais silvestres, encaminhadas e mantidas sob regime de quarentena nos centros de triagem de animais silvestres e destinadas ? soltura. Este trabalho teve por objetivo, determinar o ritmo circadiano ou periodicidade na elimina??o de oocistos de Isospora spp. em Passeriformes, assim como identificar as esp?cies do parasito encontradas e verificar a efic?cia dos anticocc?dios na profilaxia durante o per?odo de quarentena. Foram coletadas 1393 amostras fecais de aves da ordem Passeriformes pertencentes ? diversas fam?lias e esp?cies, oriundas da apreens?es do tr?fico de animais silvestres e encaminhadas ao CETAS/IBAMA, Serop?dica, RJ. Ap?s um per?odo de esporula??o, as amostras foram submetidas a t?cnica de centr?fugo-flutua??o com sacarose, quantificadas e os resultados expressos em OoPD (oocistos por defeca??o). Os resultados demonstraram que independentemente do continente onde as aves habitam, o fotoper?odo ? um fator importante na manuten??o da periodicidade da elimina??o dos oocistos de Isospora spp. e os Passeriformes de diversas fam?lias apresentaram um valor m?dio de OoPD mais elevado no per?odo da tarde. Foi verificado o controle da coccidiose nestes p?ssaros atrav?s do uso de anticocc?dios e observou-se que a efic?cia pode variar de acordo com a esp?cie do parasito e dos p?ssaros, os quais apresentam h?bitos comportamentais e alimentares diversificados que podem influenciar na resposta ao tratamento.

Submitted by Celso Magalhaes (celsomagalhaes@ufrrj.br) on 2017-05-29T13:53:42Z No. of bitstreams: 1 2016 - Lidiane Maria da Silva.pdf: 11158526 bytes, checksum: 92a272f4f09e476cee2262d286c61963 (MD5) / Made available in DSpace on 2017-05-29T13:53:43Z (GMT). No. of bitstreams: 1 2016 - Lidiane Maria da Silva.pdf: 11158526 bytes, checksum: 92a272f4f09e476cee2262d286c61963 (MD5) Previous issue date: 2016-02-18 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / The thaminophilid passerines, just as other families of Passeriformes, can be parasitized by different species of coccidia, especially the genera Isospora Schneider, 1881 and Eimeria Schneider, 1875. In this context, this study aimed to identify, characterize and quantify coccidian species from Thamnophilidae in the Itatiaia National Park. Seven expeditions were performed at Itatiaia National Park, of which five were in conserved areas and two in areas around the park. A total of 184 species of birds were captured with mist nets, being 26 thaminophilid passerines. After fecal sampling and processing were observed coccidia of the genera Isospora and Eimeria. The species Isospora parnaitatiaiensis Silva, Rodrigues, Lopes, Berto, Luz, Ferreira, Lopes, 2015 was identified in two different hosts, Pyriglena leucoptera (Vieillot, 1818) and Dysithamnus mentalis (Temminck, 1823), and their oocysts were characterized as polymorphic, since the oocysts from P. leucoptera were more ellipsoidal and the oocysts from D. mentalis were more sub-spherical, which may be the result of speciation process/adaptation to these hosts. The intensities of infection in different hosts were relatively low, since that P. leucoptera and D. mentalis shed together an OoPD of 316 oocysts of I. parnaitatiaiensis, which can be explained by the conserved environment in the Itatiaia National Park and the insectivore feeding habit. Finally, the specificity occurred at the family level, because P. leucoptera and D. mentalis, both of Thamnophilidae family, have been reported as hosts of I. parnaitatiaiensis / Os taminofil?deos, da mesma forma que outras fam?lias de Passeriformes, podem ser parasitados por diversas esp?cies de cocc?dios, principalmente dos g?neros Isospora Schneider, 1881 e Eimeria Schneider, 1875. Neste contexto, este trabalho teve por objetivo identificar, caracterizar e quantificar esp?cies de cocc?dios parasitos de Thamnophilidae do PNI. Foram realizadas sete expedi??es no Parque Nacional do Itatiaia, das quais cinco foram em ?reas mais preservadas e duas em ?reas no entorno do parque, as aves foram capturadas com o auxilio de redes de neblina, ao todos foram capturados 184 esp?cimes de aves sendo 26 taminofil?deos, ap?s o processamento das amostras observou ser cocc?dios do g?nero Isospora e Eimeria. A esp?cie Isospora parnaitatiaiensis Silva, Rodrigues, Lopes, Berto, Luz, Ferreira, Lopes, 2015 foi identificada em dois diferentes hospedeiros da fam?lia Thamnophilidae, Pyriglena leucoptera (Vieillot, 1818) e Dysithamnus mentalis (Temminck, 1823), sendo que seus oocistos foram caracterizados como polim?rficos, j? que os oocistos de P. leucoptera s?o mais elips?ides em rela??o aos oocistos de D. mentalis que tendem ao ser mais sub-esf?ricos, o que pode ser consequ?ncia do processo de especia??o/adapta??o ao hospedeiro. A intensidade de infec??o nos diferentes hospedeiros taminofil?deos positivos foram relativamente baixas, P. leucoptera e D. mentalis tiveram juntos um OoPD de 316 para os oocistos de I. parnaitatiaiensis, o que pode ser justificado pelo ambiente conservado do PNI e pelo h?bito alimentar inset?voro. Finalmente, a especificidade ocorreu em n?vel de fam?lia, pelo fato de P. leucoptera e D. mentalis, ambos da fam?lia Thamnophilidae, ter sido relatados como hospedeiros de I. parnaitatiaiensis

Feldstudie zum Einsatz gezielter Desinfektionsmaßnahmen gegen die Saugferkelkokzidiose Institut für Parasitologie der Veterinärmedizinischen Fakultät der Universität Leipzig Eingereicht im März 2004 Schlüsselworte: Saugferkelkokzidiose, Isospora suis, Bekämpfung, Desinfektion, Ne-opredisan 135-1® 109 Seiten, 13 Abbildungen, 12 Tabellen, 137 Literaturangaben, 11 Anhangstabellen Die neonatale Kokzidiose des Schweins ist ein weit verbreitetes Problem in Ferkelerzeugerbe-trieben, das üblicherweise durch medikamentelle Metaphylaxe bekämpft wird. Aufgabe der hier vorliegenden Arbeit war es, eine gezielte Desinfektion mit Neopredisan 135-1®, einem gegen Kokzidienoozysten wirksamen Desinfektionsmittel, als mögliche Alternative zur medi-kamentellen Bekämpfung zu testen. In zwei Ferkelerzeugerbetrieben im Landkreis Diepholz (Niedersachsen) mit vorberichtlicher Isosporoseproblematik wurden von Oktober 2001 bis Juli 2002 in sechs Versuchsdurchgän-gen (V1 bis V6) insgesamt 56 Würfe (Gruppe A) nach einer Eingangsdesinfektion und zu-sätzlicher Desinfektion in der belegten Abferkelbox mit Neopredisan 135-1® auf Isospora suis koproskopisch und klinisch untersucht und mit 50 Würfen, die als Kontrollgruppe (Grup-pe B) geführt wurden, verglichen. In der Versuchsgruppe (Gruppe A) wurde einmal (V1 und Gruppe A1 in V5) gegen Ende der ersten Lebenswoche, zweimal (V2, Gruppe A2 in V5, V6) gegen Ende der ersten und zweiten Lebenswoche oder dreimal (V3 und V4) gegen Ende der ersten Lebenswoche und erneut zwei und vier Tage später eine Desinfektion durchgeführt. Die Würfe wurden ab dem 5. Lebenstag siebenmal bis zum Absetzen nach der dritten Le-benswoche untersucht, wobei jeweils vier Ferkeln pro Wurf Einzelkotproben rektal entnom-men und die beprobten Ferkel klinisch beurteilt wurden. Durchfallkotproben wurden differen-tialdiagnostisch auf andere Enteropathogene (E. coli, Clostridien, Salmonellen, Lawsonia intracellularis, Brachyspira hyodysenteriae) untersucht. Am ersten und letzten Untersu-chungstermin wurden die Ferkel jeweils gewogen. Von den Muttersauen wurden am ersten und letzten Untersuchungstermin eines Versuchsdurchganges Kotproben parasitologisch un-tersucht. In V3 war aufgrund einer mangelhaft durchgeführten Reinigung erwartungsgemäß keine Des-infektionswirkung zu verzeichnen, und dieser Versuchsdurchgang wurde nicht weiter ausge-wertet. Mit Ausnahme von V6 waren die Befallsraten der Würfe mit Isospora suis in den zu-sätzlich desinfizierten Buchten geringer als bei den Kontrollen. Bezogen auf die Einzelproben waren in allen fünf ausgewerteten Versuchsdurchgängen in der Gruppe A insgesamt weniger Proben positiv als in der Kontrollgruppe B. Dabei schwankten die Befallsraten der Würfe in den einzelnen Versuchsdurchgängen zwischen 14 % und 90 % in der Gruppe A und zwischen 43 % und 83 % in der Gruppe B. Mit einer einmaligen Zwischendesinfektion war eine Reduk-tion der Prävalenz von Isospora suis um 67 % bis 77 % erreichbar. Die zweimalige Desinfek-tion reduzierte das Auftreten von I. suis um 54 %, 71 % und 83 % und die dreimalige Zwi-schendesinfektion um 76 %. Somit hat die Zwischendesinfektion zwar den Infektionsdruck senken können, die Wirkung konnte aber nicht durch eine erhöhte Frequenz der Desinfektion verbessert werden. Insgesamt hatten 7 % der Proben Durchfallcharakter. In V1, V2 und V5 waren dabei in der Gruppe A weniger Würfe betroffen als in der Kontrollgruppe B, in V4 und V6 konnte ein entsprechender Effekt nicht festgestellt werden. Die Durchfallraten der Würfe schwankten in den einzelnen Versuchsdurchgängen zwischen 43 % und 90 % in der Gruppe A und zwischen 60 % und 90 % in der Gruppe B. Bezogen auf die Ferkel war eine Reduktion des Auftretens von Durchfall außer in V2 regelmäßig erreichbar. Mit einer einmaligen Zwischendesinfektion konnte die Durchfallhäufigkeit um 43 % und 65 % reduziert werden, mit zweimaliger Desin-fektion um 18 % und 48 % und mit dreimaliger Desinfektion um 39 %. Die Durchfallhäufig-keit konnte insgesamt durch die Zwischendesinfektion reduziert werden, aber auch hier brach-te eine erhöhte Frequenz der Desinfektion keinen zusätzlichen Nutzen. Auf weitere klinische Parameter sowie die Gewichtsentwicklung der Ferkel hatte die Zwi-schendesinfektion wenig bis gar keinen Einfluss. Bei den Sauen waren in wenigen Fällen Eier von Magen-Darm-Strongyliden sowie Oozysten von Eimerien nachweisbar. Oozysten von I. suis konnten hier nicht gefunden werden. Die Ergebnisse zeigen unter Feldbedingungen einen hemmenden Einfluss von Neopredisan 135-1® auf die Ausbreitung der Isosporose, so dass in einer gezielten Desinfektion ein zur Metaphylaxe alternativer Ansatz für eine Bekämpfung liegen könnte. Allerdings ist in Be-ständen mit bestehender klinischer Problematik eine alleinige Desinfektion nicht ausreichend, so dass hier eine Kombination von medikamenteller Metaphylaxe und Desinfektion vorge-schlagen wird. Möglichkeit und Nutzen von integrierten Bekämpfungsmaßnahmen, die ein parasitologisch-klinisches Monitoring einschließen müssen, sollten in weiteren Studien unter-sucht werden. / Field study concerning the suitability of disinfection in fighting piglet coccidiosis for Institute of Parasitology, Faculty of Veterinary Medicine, University of Leipzig Submitted in March 2004 Keywords: Piglet coccidiosis, Isospora suis, Control, Disinfection, Neopredisan 135-1® 109 pages, 13 figures, 12 tables, 137 references, 11 appendices Piglet coccidiosis is a well-known and frequent problem in piglet production. It is usually fought by metaphylaxis. The objective of this investigation was to try specific disinfection with Neopredisan 135-1® for the control of coccidiosis as an alternative approach to medica-tion. Between October 2001 and July 2002, 56 litters (group A) kept in pens that were additionally disinfected after farrowing with Neopredisan 135-1® were compared to 50 litters (group B) without disinfection. The study was performed in six sequences (V1 to V6) on two piglet breeding farms with a history of Isosporosis (Diepholz, Lower Saxony). In group A, addi-tional disinfection was conducted at the end of the first week of life (V1 and group A1 in V5), a second, additional disinfection at the end of the first and second week of life (V2, group A2 in V5, V6) and three additional disinfections were made at the end of the first week of life and two and four days later (V3 and V4). Litters were examined seven times from the fifth day after farrowing until weaning. Individual faecal samples from four piglets per litter were col-lected. Each piglet was examined clinically. Subjects with diarrhea were assayed for Entero-bacteriaceae for differential diagnosis. On the first and the last examination day piglets were weighed. Samples from the sows were examined for parasites on the first and the last exami-nation day. Sequence V3 was discarded because of insufficient stall cleanliness, which, as expected, re-sulted in failure of disinfection. In general, the prevalence of Isospora suis was lower in the litters where repeated disinfections were conducted than in the litters without additional disin-fection with the exception of V6. I. suis was discovered more often in the individual samples from group B than from group A in all five trial sequences. The prevalence in the litters ranged from 14 % to 90 % in group A and from 43 % to 83 % in group B. A single additional disinfection reduced the prevalence of I. suis by 67 % to 77 %, two additional disinfections reduced the prevalence of I. suis by 54 %, 71 % and 83 % and three additional disinfections reduced the prevalence of I. suis by 76 %. Thus disinfection after farrowing is suited to reduce the infection pressure, however increased frequency of additional measures did obviously not improve the hygienic status. Diarrhea was diagnosed in 7 % of all subjects. In the sequences V1, V2 and V5, more litters in group B showed diarrhea than in group A. In the sequences V4 and V6 there were equal numbers of litters with diarrhea in both groups. The prevalence of diarrhea in the litters was ranging from 43 % to 90 % in group A and from 60 % to 90 % in group B. Related to the in-dividual piglets diarrhea was less frequently seen in group A. A single additional disinfection reduced the prevalence of diarrhea by 43 % and 65 %, two additional disinfections reduced the prevalence of diarrhea by 18 % and 48 % and three additional disinfections reduced the prevalence of diarrhea by 39 %. However, in trial sequence V2 (repeated disinfection) no re-duction of the prevalence of diarrhea was observed. Disinfection after farrowing is suited to reduce the prevalence of diarrhea, but no improvement can be seen by increased frequency of additional measures. Other clinical aspects, such as weight gain, were not influenced by additional disinfection. In the samples from the sows eggs of Strongyloides and oocysts of Eimeria were occasionally seen, oocysts of I. suis were not found. The present data show that Neopredisan 135-1® can inhibit the spread of piglet coccidiosis under field conditions. Specific disinfection may be a suitable control measure against piglet coccidiosis. In case of clinical coccidiosis, disinfection alone will not suffice but may support medical metaphylaxis. More investigations are required to explore the suitability of integrated control measures that should include parasitological monitoring.

Submitted by Pós graduação Farmácia (ppgfar@ufba.br) on 2017-05-30T20:39:16Z No. of bitstreams: 1 Flavia-Thamires -farmacia-final.pdf: 2671133 bytes, checksum: d04fe328ab695f2547b14e8d937e5959 (MD5) / Approved for entry into archive by Patricia Barroso (pbarroso@ufba.br) on 2017-06-01T17:33:52Z (GMT) No. of bitstreams: 1 Flavia-Thamires -farmacia-final.pdf: 2671133 bytes, checksum: d04fe328ab695f2547b14e8d937e5959 (MD5) / Made available in DSpace on 2017-06-01T17:33:52Z (GMT). No. of bitstreams: 1 Flavia-Thamires -farmacia-final.pdf: 2671133 bytes, checksum: d04fe328ab695f2547b14e8d937e5959 (MD5) / FAPESB / O diagnóstico dos coccídeos Cryptosporidium e Isospora belli é realizado principalmente pela pesquisa de oocistos em esfregaços fecais corados. Entretanto, não existe uma padronização na rotina laboratorial para a identificação microscópica desses coccídeos. O diagnóstico da Cryptosporidium pode também ser realizado pela detecção de coproantígenos por ensaio imunoenzimático (ELISA) ou pela amplificação do DNA através da reação em cadeia da polimerase (PCR), dispensando a identificação morfológica dos oocistos. Além disso, a análise do polimorfismo genético de fragmentos de restrição (PCR-RFLP) pode ser utilizada na caracterização das espécies de Cryptosporidium, permitindo um melhor entendimento da dinâmica da transmissão deste parasito. Os objetivos deste trabalho foram: (1) comparar as técnicas de concentração de formol-acetato de etila (FE) e sedimentação por centrifugação (SC), bem como as técnicas de coloração de Ziehl-Neelsen modificado (ZN), auramina (AR) e safranina (SF) na detecção de oocistos de Cryptosporidium e Isospora belli em amostras fecais; (2) comparar a microscopia com a pesquisa de coproantígeno para o diagnóstico de Cryptosporidium e avaliar os resultados discordantes utilizando a PCR e (3) caracterizar as espécies de Cryptosporidium de amostras fecais humanas, através da PCR-RFLP. Para comparação entre os métodos de concentração de oocistos, FE e SC, e as técnicas de coloração, ZN, AR e SF, foram utilizadas amostras fecais positivas para Cryptosporidium (n=27) e I. belli (n=15), conservadas em formalina a 10%. Os métodos foram avaliados quanto ao número de oocistos detectados e a qualidade microscópica dos esfregaços. Para comparação entre o método de ZN e ELISA para o diagnóstico de Cryptosporidium foram examinadas amostras fecais de 626 crianças de diferentes grupos. Posteriormente, todas as amostras positivas para Cryptosporidium obtidas no estudo, juntamente com outros isolados disponíveis no laboratório, foram submetidos à extração de DNA e análise por Nested-PCR/RFLP dos genes COWP e 18S rRNA, para determinação das espécies de Crptosporidium. Os métodos SC e ZN identificaram mais oocistos de ambos parasitos do que os demais métodos avaliados (p<0,05). Houve perda de oocistos no anel de detritos gordurosos no FE em praticamente todas as amostras de Cryptosporidium e I. belli. Por outro lado, os métodos FE e AR apresentaram menos artefatos nos esfregaços comparados aos demais, sendo classificados com qualidade microscópica superior. A frequência de Cryptosporidium nas crianças foi de 2,6% (16/626), sendo maior no grupo com doença diarreica, enfatizando a importância deste coccídeo como agente etiológico da diarreia infantil. A sensibilidade e especificidade do ELISA foram de 85,7% e 99,7%, respectivamente. A eficiência da amplificação do DNA de Cryptosporidium foi de 92% (23/25), considerando os resultados dos dois genes analisados. As espécies do parasito identificadas foram C. hominis (78,3%), C. felis (8,7%), C. parvum (4,3%) e mistura C. felis + C. hominis (8,7%). Esses dados são os primeiros de genotipagem de Cryptosporidium de indivíduos de Salvador, demonstrando a predominância de C. hominis nas infecções, e a elevada frequência de C. felis, sugerindo um provável papel de gatos na transmissão do parasito aos humanos em nosso meio. / The diagnosis of coccidia Isospora belli and Cryptosporidium is usually accomplished by identification of oocysts in stained fecal smears. However, there is a lack of standardization in the routine laboratory for microscopic identification of these coccidia. The diagnosis of Cryptosporidium can also be done by detecting coproantigens using the enzyme linked immunosorbent assay (ELISA) or by parasite DNA amplification by polymerase chain reaction (PCR), eliminating the need of morphological identification of oocysts. Furthermore, the analysis of restriction fragment length polymorphism of amplified DNA (RFLP-PCR) can be used to characterize the species of Cryptosporidium, allowing a better understanding of the transmission dynamics of the parasite. The objectives of this study were: (1) to compare the techniques of concentration of formalin-ethyl acetate (FE) and sedimentation by centrifugation (SC), as well as the techniques of modified Ziehl-Neelsen (ZN), auramin (AR) and safranin (SF) in the detection of Cryptosporidium and Isospora belli in stool samples, (2) to compare the microscopy with coproantigen detection for the diagnosis of Cryptosporidium and evaluate discordant results using PCR and (3) to characterize the species Cryptosporidium in human fecal samples by PCR-RFLP. To compare the methods for concentration, FE and SC, and staining of oocysts, ZN, AR and SF, there were used positive fecal samples for Cryptosporidium (n = 27) and I. belli (n = 15), preserved in 10% formalin. The methods were evaluated according the numbers of oocysts detected and the microscopic quality of smears. For comparison between ELISA and ZN for the diagnosis of Cryptosporidium in faecal samples, there were examined 626 stool samples from children of different groups. Subsequently, all positive samples for Cryptosporidium obtained in the study, along with other isolates available in the laboratory, were subjected to DNA extraction and analysis by Nested-PCR/RFLP of the COWP and 18S rRNA genes to determine the species of Crptosporidium. The SC and ZN methods identified more oocysts of both parasites than other methods tested (p <0.05). The loss of oocysts in the fatty debris layer of FE method was observed in all Cryptosporidium and I. belli samples. Moreover, the methods of FE and AR had fewer artifacts in smears compared to the others, being ranked with higher microscopic quality. The frequency of Cryptosporidium in children was 2.6% (16/626), being higher in patients with diarrheic disease, emphasizing the importance of this protozoan as etiologic agent of childhood diarrhea. The sensitivity and specificity of ELISA were 85.7% and 99.7%, respectively. The efficiency of amplification of Cryptosporidium DNA was 92% (23/25), considering the results of the two genes. The species of the parasite identified were C. hominis (78.3%), C. felis (8.7%), C. parvum (4.3%) and a mixture of C. felis + C. hominis (8.7%). To our knowledgment, these data represent the first genotyping study of Cryptosporidium from individuals of Salvador, demonstrating the dominance of C. hominis infection and the high frequency of C. felis, suggesting a potential role of cats in the transmission of the parasite to humans in our area.

In two years of observation, (spring 2006, autumn 2006, spring 2007 and autumn 2007) were being screened for parasites in total 495 faecal samples coming out of three farms from Ceske Budejovice (285 samples of sucking pigs not older then 28 days and 174 samples of piglets not older then 8 weeks). The method used to examine those samples was a flotation-concentrating method (Sheather{\crq}s carbohydrate fusion) and in the year of 2007 was also used a specific aniline-carbol-methyl violet staining method to detect the Cryptosporidium spp. followed by positive molecular characterized (direct sequencing of partial SSU rRNA partial genes and PCR-RFLP at the SSU rRNA). In screened samples were mainly detected parasites named Cryptosporidium spp., found in 4,1% of faecal samples in 2006 and in 2007 in 32,8% faecal samples, out of which 14,4% was found in pre-weaned piglets samples and 26,4% in post-weaned piglets. Based on genotyping provided on positive samples out of the year 2007, using method of sequensing analysis SSU rRNA, was the occurence of C. suis, Cryptosporidium pig genotype II aC. Muris described. High prevalence of Isosporou suis 13,9 % (64/459) was also detected with its appearance, in particular, in pre-weaned piglets 21,4 % (61/285). Further on, some of other identified group was Eimerie spp. 5,7 % (26/459) infecting, in the main, post-weaned piglets 10,9% (19/174) and Giardia intestinalis 2,4 % (10/459). Most of the samples mentioned occured in conditioned faeces and there is no seasonal relationship to the parasital occurance.