Este estudo teve como objetivo a padronização de protocolos e a seleção de novos primers para a identificação espécie-específica de Taenia saginata e Taenia solium através da reação em cadeia da polimerase (PCR) e duplex-PCR. Inicialmente, foram recuperadas seqüências depositadas no GenBank (acesso n° AB020399 para T. saginata e n° AB020395 para T. solium) referentes ao gene da subunidade maior do ribossomo (LSU RNAr) de tenídeos. A partir do alinhamento das seqüências, um primer genérico denominado TBR-3 (5'-ggcttgtttgaatggtttgacg- 3') foi selecionado de região conservada e, de diferentes regiões semi-conservadas, os primers específicos TBR-4 para T. saginata (5'-cgactcatgaagataaacaaggt-3') e TBR-5 (5'-cggtcgaacagaccataaatct-3') e TBR-6 (5'-gctactacacctaaattctaacc- 3') para T. solium. Os primers foram avaliados quanto à especificidade através da PCR empregando-se DNA total (DNAt) de amostras de cisticercos e proglotes dos parasitos, previamente identificadas por critérios morfológicos. O par de primers TBR-3/TBR-4 permitiu a amplificação específica do fragmento esperado de 328 pb a partir do DNAt de T. saginata. Os pares TBR-3/TBR-5 e TBR-3/TBR-6 permitiram a amplificação, respectivamente, dos fragmentos específicos de 310pb e 286pb a partir do DNAt de T. solium. A identidade dos produtos de PCR foi comprovada comparando-se a seqüência dos amplicons obtidos às seqüências de referência do gene LSU RNAr registrado no GenBank (n° AB020399 e n° AB020395). As reações apresentaram sensibilidade para detecção de até 1fg do DNAt de T. solium e 0,2fg do DNAt de T. saginata. A combinação dos primers TBR-3/TBR-4 e TBR3/TBR-6 e o tamanho dos fragmentos gênicos obtidos permitiram o estabelecimento de ensaios de duplex-PCR, eficaz na detecção simultânea do DNA de T. saginata e T. solium em sistema único de reação. Os primers utilizados não geraram qualquer produto de amplificação cruzada quando testados com DNAt de Taenia hydatigena, Taenia taeniaeformis, Hymenolepis diminuta, Anoplocephala magna, Paranoplocephala mamillana e Moniezia expansa, nem frente ao DNAt dos hospedeiros Homo sapiens, Bos taurus e Sus scrofa.

This study was conducted to evaluate a protocol and to select novel primers for the species-specific identification of Taenia saginata and Taenia solium by PCR and duplex-PCR assays. Sequences of the LSU rRNA gene of taenids were obtained from the GenBank (T. saginata access n° AB020399 and T. solium access n° AB020395). The sequences were aligned and then used for primer design. The generic primer TBR3 (5'-ggcttgtttgaatggtttgacg- 3') was selected from a conserved region. The T. saginata specific primer TBR-4 (5'-cgactcatgaagataaacaaggt-3') as well as T. solium specific primers TBR-5 (5'-cggtcgaacagaccataaatct-3') and TBR-6 (5'-gctactacacctaaattctaacc- 3') were selected from different semi-conserved regions. The selected sequences were examined in for similarities with other organisms through the GenBank Blast procedure and experimentally by PCR using total DNA (tDNA) extracted from cysticerci and proglottids from both parasites. The primer pair TBR-3/TBR-4 amplified specific fragments of 328 bp from T. saginata tDNA. The pairs TBR-3/TBR5 and TBR-3/TBR-6 amplified, respectively, the expected and specific fragments of 310bp and 286bp from the T. solium tDNA. Sequencing of the amplicons followed by comparison to GenBank reference sequences confirmed the identities of the PCR products. The detection sensitivity was equivalent to 1fg of T. solium tDNA and 0,2fg of T. saginata tDNA. The combination of primers TBR-3/TBR-4 and TBR3/TBR-6 and the size of amplicons allowed the establishment of a duplex-PCR assay to detect T. saginata and T. solium DNA. No cross reaction was observed with any combination of primers in reactions with tDNA of the parasites Taenia hydatigena, Taenia taeniaeformis, Hymenolepis diminuta, Anoplocephala magna, Paranoplocephala mamillana and Moniezia expansa, nether from the hosts tDNA Homo sapiens, Bos taurus nor Sus scrofa.

Relata-se o encontro de um novo caso de bertiellose humana em paciente do sexo feminino, 2 anos, natural de Goiânia, Estado de Goiás com histórico de permanência em área freqüentada por símios no Estado de Mato Grosso. A paciente apresentava inapetência, dores abdominais, emagrecimento e eliminava nas fezes ovos e proglotes que foram identificados como Bertiella sp. O objetivo do presente trabalho é registrar o encontro do terceiro caso de parasitismo humano por Bertiella sp. (Cestoda-Anoplocephalidae) no Brasil, caracterizando mais um caso de zoonose helmíntica.

The authors report on a new case of human Bertiellosis in a 2-year old female patient who was born in Goiânia-Goiás (Brazil) and has had history of permanent dwelling in an area frequently visited by simians in Mato Grosso (Brazil). At the time of diagnosis the patient showed inappetence, abdominal pain, and loss of weight. Eggs and proglottids were found in her stool and were identified as Bertiella sp. The objective of this report is to register the third case of human Bertiellosis in Brazil, characterizing one more case of helminthic zoonosis.

With the purpose of investigating the biological aspects of Angiostrongylus vasorum (Baillet, 1866) Kamensky, 1905 in Subulina octona Bruguiere, 1789 experimental infections were carried out with the exposure of 90 adult molluscs to 155 L1 of the parasite/mollusc during 24 hours. The intramollus-can larval development was checked out daily through the chemical digestion of four infected molluscs and one molluscs from the control group during 21 days after infection. Until the 6th day post-infection, only Li of the parasite has been observed. On the 7th day; the presence of L2 was detected and from the 13th day on, L3 of A. vasorum also appeared. By means of larval morphometry, it has been observed that, Li had na average size of 335 x 14,5 um whereas L2 had an average size of 440,7 x 33,9 um on the 7th day ofter infection, reaching a, aximal diameter of 41,8 um on the 12th day after infections. The L3 had an average size of 472,4 x 38,7 um on the 13th day day and on the 21st' day after infection the measure was 551,3 x 27,6 um. In order to verify the infectivity of L3, from 5. octona to the definitive host, infection of a dog was undertaken by means of ingestion of tritured molluscs. On the 49th day post-infection, Li was found in the feaces of the dog and at necropsy on the 84th day post-infection adults worms were found in the lungs.

Self-fertilization in Subulina octona (Bruguiére, 1789) was observed among 32 snails. Individually isolated for 154 days. Eggs were dctected in 38 to 50 days old snails; total number of ovipositions per snail varied from six-nine; number of eggs per cluster, one-nine and total number of eggs per snail, 16-56 during the period of study. A period of incubation of one to 15 days and a viability of 94.8% were observed for eggs layed by these isolated snails.