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Thesis--University of California at Berkeley. / eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 256-282).

Thesis--University of California at Berkeley. / Includes bibliographical references (leaves 256-282). Also issued in print.

Made available in DSpace on 2014-06-12T17:35:11Z (GMT). No. of bitstreams: 2 arquivo4409_1.pdf: 676822 bytes, checksum: 4a1153b3912f6483c79f20c422b118f1 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2002 / Oligonucleotídeos foram construídos com base na sequência primária do gene codificando o rRNA de Plasmodium para amplificar DNA de P. falciparum, P. vivax, P. malariae e P. ovale, de maneira gênero-específica. Três sistemas de PCR foram utilizados: PCR simples, hemi-nested PCR convencional e hemi-nested PCR em um único tubo, desenvolvidos em nosso laboratório. Na PCR simples, composta de 30 ciclos, foram utilizados os oligonucleotídeos GJ1 e HR842 (20 pmol/50μl), já testados por nosso grupo. Na hemi-nested PCR convencional utilizou-se três oligonucleotídeos (GJ1, PGFO3 e HR842), em duas reações sequenciais, sendo o PGFO3 construído durante o desenvolvimento do presente trabalho, visando a detecção do gênero Plasmodium. O par GJ1 e HR842 foram utilizados como oligonucleotídeos externos na primeira reação, e o PGFO3 como interno, ancorado ao HR842 na segunda reação. A hemi-nested PCR em um único tubo consistiu em 60 ciclos (92ºC, 30s; 58ºC, 30s e 72ºC, 45s), e concentrações limitantes de oligonucleotídeos externos (4 pmols/50μl) participavam da PCR sem competição com os oligonucleotídeos internos durante os primeiros 15 ciclos da reação e 40 pmol/50μl de primers internos (imobilizados na face interna da tampa do microtubo) foram introduzidos na PCR no 16º ciclo. As concentrações dos outros componentes da reação foram as mesmas utilizadas nas reações convencionais de PCR. Observou-se que a quantidade mínima de DNA genômico detectada pela PCR simples, hemi-nested PCR e hemi-nested PCR em um único tubo foi de 10 pg; 0,01 pg e 0,1 pg, respectivamente. Apesar da hemi-nested PCR em um único tubo ter sido menos sensível que a heminested PCR convencional, é muito mais simples e conveniente, já que o risco de contaminação cruzada é bem menor. Esses sistemas moleculares de diagnóstico podem ser usados em situações quando se requer uma alta sensibilidade e especificidade, tais como na avaliação da eficiência de quimioterapia, detecção precoce de infecção e prevenção de transmissão a partir de pacientes hipoparasitêmicos

Retortamonads (Retortamonadida; genera Chilomastix and Retortamonas) are a small group of protists comprising intestinal commensals of both vertebrates and invertebrates and one free-living species of the genus Chilomastix. Molecular phylogenetic studies showed that retortamonads are closely related to diplomonads, Carpediemonas, Dysnectes, Hicanonectes, Kipferlia and two undescribed lineages of free-living Carpediemonas-like organisms, together forming the monophyletic excavate group Fornicata. For a long time Retortamonadida have been assumed to be a monophyletic group. However, first molecular phylogenetic study including sequence data from both Retortamonas and Chilomastix suggested that Retortamonadida are paraphyletic and that diplomonads branch within Retortamonadida. Retortamonads still remain poorly studied protist group and their phylogeny is unclear. We sequenced and analysed SSU rDNA of ten Retortamonas and four Chilomastix SSU rDNA sequences. In addition, we sequenced SSU rDNA of an undescribed enteromonad lineage. The phylogenetic tree of Fornicata was largely unresolved and the phylogenetic position of the genus Chilomastix remained unclear. On the other hand, the genus Retortamonas and diplomonads formed a robust clade. Retortamonas sequences split into three host-specific lineages. The...

Thesis (M.A.)--University of Hong Kong, 1991. / Added title in Chinese. Also available in print.

Population structure and phylogenetic relationships were studied in coccidia parasitizing the rodent families Muridae and Arvicolidae, in 40 localities in 14 European countries. Sequences of mitochondrial gene for cytochrome c oxidase subunit I (COI) and nuclear 18S rRNA gene (SSU) were used for phylogenetic analyses and for reconstruction of evolutionary relationships among coccidian species.

The prevalence and molecular characterization of Cryptosporidium spp. in slaughtered cattle 6 months and older was performed. Three species of Cryptosporidium were identified. A subtype of C. parvum was obtained.

Archamoebae is a small group of anaerobic protists belonging to the eukaryotic supergroup Amoebozoa. Historically, they were regarded as primitively amitochondriate. However, a mitochondrial remnant has been found in some archamoebae. Phylogenetic analyses showed that Archamoebae are closely related to the aerobic slime moulds (Mycetozoa). Trophozoites of archamoebae are amoeboflagellates or aflagellated amoebae. The group includes both parasitic (Entamoeba, Endolimax and, possibly, Endamoeba and Iodamoeba) and free-living (Mastigamoeba, Mastigella, Pelomyxa) genera. The genus Mastigina comprises both endozoic and free-living representatives. Flagellated genera Mastigina, Mastigamoeba, Mastigella and Pelomyxa possess a single basal body associated with a microtubular cone which may or may not be associated with nucleus. The cone is a common feature for Archamoebae and mycetozoan slime moulds. The phylogeny of Archamoebae has not been fully elucidated yet and the taxonomy of free-living representatives is confusing. In the present study, we obtained 42 stable isolates of free-living Archamoebae. We sequenced and analyzed SSU rDNA of 15 of them. The Archamoebae split into five lineages. Based on TEM, we were able to recognize genera Mastigamoeba and Mastigella. The isolate IND8 probably represents a new...

University of Macau / Institute of Chinese Medical Sciences