Avaliação e retificação da identificação específica de moluscos do gênero Biomphalaria PRESTON, 1910 do acervo da Coleção de Malacologia Médica (Fiocruz-CMM)

Submitted by Nuzia Santos ([email protected]) on 2012-08-17T13:05:38Z No. of bitstreams: 1 Dissertacao_Cryslaine Aguiar Silva[1].pdf: 4463723 bytes, checksum: 12a5258b0c98c7c52fdd0066ba8b4910 (MD5)Made available in DSpace on 2012-08-17T13:05:38Z (GMT). No. of bitstreams: 1 Dissertacao_Cryslaine Aguiar Silva[1].pdf: 4463723 bytes, checksum: 12a5258b0c98c7c52fdd0066ba8b4910 (MD5)Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Belo Horizonte, MG, Brasil.As coleções biológicas constituem importante fonte de informações, propiciando conhecimento e desenvolvimento científico. A Coleção de Malacologia Médica (Fiocruz-CMM) do Centro de Pesquisas René Rachou, localizada no Laboratório de Helmintologia e Malacologia Médica (LHMM) conta atualmente com cerca de onze mil exemplares de moluscos de importância médica e veterinária, principalmente os do gênero Biomphalaria. Desde a década de 1990, o LHMM recebe moluscos de diversas regiões do Brasil e do exterior e utiliza caracteres morfológicos e/ou moleculares na identificação das espécies. Parte do acervo não foi identificada por técnicas moleculares e em 332 pontos de coleta os exemplares encontravam-se sem identificação. Assim, o objetivo do presente trabalho foi avaliar e retificar a identificação dos moluscos do gênero Biomphalaria presentes na Fiocruz- CMM. Para isso o acervo foi separado em dois grupos: grupo I: exemplares inseridos entre 1993 e 2002, com 620 pontos de coleta e 5.137 exemplares; grupo II: inseridos entre 2003 e 2009, com 464 pontos de coleta e 1.924 exemplares. A identificação específica dos exemplares do acervo foi realizada pela morfologia e/ou pela PCR-RFLP. Os resultados foram agrupados em seis categorias: 1) dados corretos; 2) equívocos (classificados como outra espécie); 3) adequações (espécie em sinonímia); 4) inconclusivos (mesma espécie com perfis moleculares diferentes), 5) material degradado e 6) exemplares identificados pela primeira vez (depositados sem identificação). No grupo I, 41,8% dos pontos de coleta os exemplares estavam com dados taxonômicos corretos; em 2,1% tinham equívocos; em 0,8% houve adequação; em 25,5% a identificação foi inconclusiva; em 1,9% estavam degradados impossibilitando a identificação e em 27,9% foram identificados pela primeira vez. No grupo II, 70,7% dos pontos de coleta os exemplares estavam com dados taxonômicos corretos; em 5,6% tinham equívocos; em 10,1% a identificação foi inconclusiva; em 5,4% os exemplares estavam degradados impossibilitando a identificação e em 8,2% foram identificados pela primeira vez. Os equívocos ocorreram entre: B. peregrina e B. tenagophila (35,8%), B.tenagophila e B. glabrata (20,5%), B. tenagophila e B. occidentalis (7,7%), B. peregrina e B.intermedia (5,2%), B. intermedia e B. straminea (5,2%), B. amazonica e B. cousini (5,1%), B.straminea e B. occidentalis (2,6%), B. straminea e B. oligoza (2,6%), B. prona e B. kuhniana (2,6%), B. tenagophila e B. t. guaibensis (2,6%), B. straminea e B. glabrata (2,6%), B. tenagophila e B. straminea (2,6%), B. peregrina e B. schrammi (2,6%), e B. peregrina e B. straminea (2,6%). A adequação foi em virtude de B. obstructa e B. temascalensis serem sinonímias de B. havanensis. Na categoria inconclusiva ficaram B. aff. straminea, B. tenagophila oriundas da Argentina e B. peregrina. Estes dados demonstram a importância da utilização de mais de uma técnica na identificação específica e da boa preservação dos exemplares do acervo. Estes dados foram utilizados para atualizar os livros de registro de recebimento de moluscos, livro de tombo e a base de dados do CRIA. Estudos taxonômicos serão realizados com as espécies da categoria inconclusivos.The biological collections are an important source of information providing knowledge and scientific development. The Medical Malacology Collection (Fiocruz-CMM) of René Rachou Research Center located at the Laboratory of Medical Helminthology and Malacology (LHMM) has currently about eleven thousand specimens of molluscs of medical and veterinary importance, especially the genre Biomphalaria. Since 1990s, the LHMM receives molluscs from various regions of Brazil and abroad and utilizes morphological and/or molecular method for species’ identification. Part of the collection has not been identified by molecular techniques and 332 collection points were unidentified. Thus, o objective of this present study was assess and rectify the identification of the mollusks of genre Biomphalaria of the Fiocruz-CMM. For this the collection was separated into two groups: group I: specimens inserted between 1993 and 2002 with 620 collection points and 5.137 specimens; group II: inserted between 2003 and 2009, with 464 collection points and 1.924 specimens. The specific identification of specimens from the collection was performed by morphology and/or PCR-RFLP. The results were grouped into six categories: 1) correct data, 2) errors (classified as other species), 3) adjustments (species in synonymy), 4) inconclusive (same species with different molecular profiles) 5) degraded material and 6) sample identified to first time (deposited without identification). In group I, 41.8% of collection points were with correct taxonomic data, 2.1% had errors; 0.8% was adjustment; 25.5% the identification was inconclusive, 1.9% were degraded that was impossible to identify and 27.9% were first time identified. In group II, 70,7% of collection points the specimens were with correct taxonomic data; 5,6% had errors; 10.1% was inconclusive, 5.4% of specimens were degraded making it impossible to identification and 8.2% of specimens were first time identified. The errors has occurred between: B. peregrina and B. tenagophila (35,8%), B. tenagophila and B. glabrata (20.5%), B. tenagophila and B. occidentalis (7.7%), B. peregrina and B. intermedia (5.2%),B. intermedia and B. straminea (5.2%), B. amazonica and B. cousini (5.1%), B. straminea and B. occidentalis (2.6%), B. straminea and B. oligoza (2.6%), B. prona and B. kuhniana (2.6%), B. tenagophila and B. t. guaibensis (2.6%), B. straminea and B. glabrata (2.6%), B. tenagophila and B. straminea (2.6%), B. peregrina and B. schrammi (2.6%), and B. peregrina and B. straminea (2.6%). The adjustment was from B. obstructa and B. temascalensis are synonymy of B. havanensis. The inconclusive category was B. aff. straminea, B. tenagophila from Argentina and B. peregrina. These data demonstrate the importance of using more than one technique in taxonomic confirmation and the good preservation of specimens’ collection. These data was utilized to update the books receive mollusks and data of CRIA. The studies will continue in specimens of the inconclusive category

Prevalence and distribution of Angiostrongylus cantonensis (Nematoda, Angiostrongylidae) in Achatina fulica (Mollusca, Gastropoda) in Baixada Santista, São Paulo, Brazil

ABSTRACT INTRODUCTION Angiostrongylus cantonensis is causes eosinophilic meningoencephalitis in humans. Worldwide expansion of this nematode is linked to the dispersion of their hosts. This study aimed to determine the prevalence of A. cantonensis infection in Achatina fulica in the nine municipalities that make up Baixada Santista, São Paulo, Brazil. METHODS Angiostrongylus cantonensis larvae were analyzed using optical microscopy. We performed polymerase chain reaction and restriction fragment length polymorphism using restriction endonuclease ClaI, directed to the internal transcribed spacer region 2 of A. cantonensis larval DNA. RESULTS Of the 540 snails analyzed, 117 (21.7%) were infected by A. cantonensis. For morphological and morphometric analyses, 60 larvae were used. Second-stage larvae were, on average, 358.2µm long and 26.4µm wide, while third-stage larvae were, on average, 450µm long and 21.12µm wide. The tails of the larvae ended in a fine tip. CONCLUSIONS All municipalities comprising Baixada Santista had A. fulica that were naturally infected with A. cantonensis. All of the observed characteristics were typical of the species

Prevalence and distribution of Angiostrongylus cantonensis (Nematoda, Angiostrongylidae) in Achatina fulica (Mollusca, Gastropoda) in Baixada Santista, São Paulo, Brazil

ABSTRACT INTRODUCTION Angiostrongylus cantonensis is causes eosinophilic meningoencephalitis in humans. Worldwide expansion of this nematode is linked to the dispersion of their hosts. This study aimed to determine the prevalence of A. cantonensis infection in Achatina fulica in the nine municipalities that make up Baixada Santista, São Paulo, Brazil. METHODS Angiostrongylus cantonensis larvae were analyzed using optical microscopy. We performed polymerase chain reaction and restriction fragment length polymorphism using restriction endonuclease ClaI, directed to the internal transcribed spacer region 2 of A. cantonensis larval DNA. RESULTS Of the 540 snails analyzed, 117 (21.7%) were infected by A. cantonensis. For morphological and morphometric analyses, 60 larvae were used. Second-stage larvae were, on average, 358.2µm long and 26.4µm wide, while third-stage larvae were, on average, 450µm long and 21.12µm wide. The tails of the larvae ended in a fine tip. CONCLUSIONS All municipalities comprising Baixada Santista had A. fulica that were naturally infected with A. cantonensis. All of the observed characteristics were typical of the species

Evaluation and updating of the Medical Malacology Collection (Fiocruz-CMM) using molecular taxonomy

Submitted by Nuzia Santos ([email protected]) on 2015-03-12T18:07:43Z No. of bitstreams: 1 2014_148.pdf: 386123 bytes, checksum: 4c373e4fe15787125ee41dcdfafac304 (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-03-12T18:07:52Z (GMT) No. of bitstreams: 1 2014_148.pdf: 386123 bytes, checksum: 4c373e4fe15787125ee41dcdfafac304 (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2015-03-12T18:11:36Z (GMT) No. of bitstreams: 1 2014_148.pdf: 386123 bytes, checksum: 4c373e4fe15787125ee41dcdfafac304 (MD5)Made available in DSpace on 2015-03-12T18:11:36Z (GMT). No. of bitstreams: 1 2014_148.pdf: 386123 bytes, checksum: 4c373e4fe15787125ee41dcdfafac304 (MD5) Previous issue date: 2014Fundação Oswald Cruz. Centro de Pesquisas René Rachou. Laboratório de Helmintologia e Malacologia Médica. Belo Horizonte, MG, Brasil.Fundação Oswald Cruz. Centro de Pesquisas René Rachou. Laboratório de Helmintologia e Malacologia Médica. Belo Horizonte, MG, Brasil/Pontifícia Universidade Católica de Minas Gerais. Belo Horizonte, MG, Brasil.Fundação Oswald Cruz. Centro de Pesquisas René Rachou. Laboratório de Helmintologia e Malacologia Médica. Belo Horizonte, MG, Brasil.Fundação Oswald Cruz. Centro de Pesquisas René Rachou. Laboratório de Helmintologia e Malacologia Médica. Belo Horizonte, MG, Brasil.Fundação Oswald Cruz. Centro de Pesquisas René Rachou. Laboratório de Helmintologia e Malacologia Médica. Belo Horizonte, MG, Brasil.Fundação Oswald Cruz. Centro de Pesquisas René Rachou. Laboratório de Helmintologia e Malacologia Médica. Belo Horizonte, MG, Brasil.Background: The Medical Malacology Collection (Coleção de Malacologia Médica, Fiocruz-CMM) is a depository of medically relevant mollusks, especially from the genus Biomphalaria, which includes the hosts of Schistosoma mansoni. Taxonomic studies of these snails have traditionally focused on the morphology of the reproductive system. However, determination of some species is complicated by the similarity shown by these characters. Molecular techniques have been used to try to overcome this problem. Description: The Fiocruz-CMM utilizes morphological and/or molecular method for species’ identification. However, part of the collection has not been identified by molecular techniques and some points were unidentified. The present study employs polymerase chain reaction-based analysis of restriction fragment length polymorphisms (PCR-RFLP) to evaluate the identification of Biomphalaria in the Fiocruz-CMM, correct existing errors, assess the suitability of taxonomic synonyms, and identify unknown specimens. The results indicated that 56.7% of the mollusk specimens were correctly identified, 4.0% were wrongly identified, and 0.4% was identified under taxonomic synonyms. Additionally, the PCR-RFLP analysis identified for the first time 17.6% of the specimens in the Collection. However, 3.1% of the specimens could not be identified because the mollusk tissues were degraded, and 18.2% of the specimens were inconclusively identified, demonstrating the need for new taxonomic studies in this group. Conclusion: The data was utilized to update data of Environmental Information Reference Center (CRIA). These studies demonstrate the importance of using more than one technique in taxonomic confirmation and the good preservation of specimens’ collection