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

Aguiar-Silva C, Mendonça CL, da Cunha Kellis Pinheiro PH, Mesquita SG, Carvalho Odos S, Caldeira RL - Springerplus (2014)

Bottom Line: 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.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.These studies demonstrate the importance of using more than one technique in taxonomic confirmation and the good preservation of specimens' collection.

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Helmintologia e Malacologia Médica do Centro de Pesquisas René Rachou-Fiocruz, Av. Augusto de Lima 1715, Belo Horizonte, MG 30190-002 Brasil.

ABSTRACT

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.

No MeSH data available.


Biomphalariarestriction profiles: 6% polyacrylamide gel showing the PCR-RFLP profiles obtained by digesting the rDNA ITS region ofBiomphalariamollusks withDdeI (lanes 1-9) andAluI (lanes 10-14). Lane 1: Biomphalaria straminea (Minas Gerais, Brazil); 2: Biomphalaria intermedia (Minas Gerais, Brazil); 3: Biomphalaria straminea (Corrientes, Argentina); 4: Biomphalaria straminea (Espinillar, Uruguay); 5: Biomphalaria tenagophila (Minas Gerais, Brazil); 6-7: Biomphalaria tenagophila (Corrientes, Argentina); 8: Biomphalaria tenagophila guaibensis (Rio Grande do Sul, Brazil); 9: Biomphalaria occidentalis (Minas Gerais, Brazil); 10: Biomphalaria tenagophila (Minas Gerais, Brazil); 11-12: Biomphalaria tenagophila (Corrientes, Argentina); 13: Biomphalaria tenagophila guaibensis (Rio Grande do Sul, Brazil); 14: Biomphalaria occidentalis (Minas Gerais, Brazil). Values to the left correspond to molecular weights in base pairs (bp).
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Fig1: Biomphalariarestriction profiles: 6% polyacrylamide gel showing the PCR-RFLP profiles obtained by digesting the rDNA ITS region ofBiomphalariamollusks withDdeI (lanes 1-9) andAluI (lanes 10-14). Lane 1: Biomphalaria straminea (Minas Gerais, Brazil); 2: Biomphalaria intermedia (Minas Gerais, Brazil); 3: Biomphalaria straminea (Corrientes, Argentina); 4: Biomphalaria straminea (Espinillar, Uruguay); 5: Biomphalaria tenagophila (Minas Gerais, Brazil); 6-7: Biomphalaria tenagophila (Corrientes, Argentina); 8: Biomphalaria tenagophila guaibensis (Rio Grande do Sul, Brazil); 9: Biomphalaria occidentalis (Minas Gerais, Brazil); 10: Biomphalaria tenagophila (Minas Gerais, Brazil); 11-12: Biomphalaria tenagophila (Corrientes, Argentina); 13: Biomphalaria tenagophila guaibensis (Rio Grande do Sul, Brazil); 14: Biomphalaria occidentalis (Minas Gerais, Brazil). Values to the left correspond to molecular weights in base pairs (bp).

Mentions: Three groups were prominent among the inconclusively identified specimens. 1) Specimens from the provinces of Corrientes, Argentina and Espinillar, Uruguay (four collection points) were morphologically similar to B. straminea, but their restriction profile for the enzyme DdeI differed from that of B. straminea (which had been previously established), here represented by sample from Minas Gerais, Brazil (Figure 1). 2) Specimens from the provinces of Corrientes, Argentina and Espinillar, Uruguay (six collection points) were morphologically similar to B. tenagophila from Brazil, but their restriction profiles for the enzymes DdeI and AluI were similar to those of B. t. guaibensis from Rio Grande do Sul, Brazil (Figure 1). 3) Specimens from 208 collection points in various Brazilian states were morphologically identified as B. peregrina, but although some of these specimens had molecular profiles characteristic of that species, others had the molecular profile of B. oligoza (which had been previously established), here represented by sample from Rio Grande do Sul, Brazil (Figure 2). In an attempt to clarify the identity of these specimens, a portion of the mitochondrial cytochrome c oxidase subunit I (COI) gene was amplified by PCR, and the restriction enzymes ClaI, RsaI, and AluI were used to generate PCR-RFLP profiles. There was no restriction site for the enzymes ClaI and RsaI. For AluI, the specimens morphologically identified as B. peregrina sometimes showed B. peregrina profiles and sometimes showed B. oligoza profiles (data not shown).Figure 1


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

Aguiar-Silva C, Mendonça CL, da Cunha Kellis Pinheiro PH, Mesquita SG, Carvalho Odos S, Caldeira RL - Springerplus (2014)

Biomphalariarestriction profiles: 6% polyacrylamide gel showing the PCR-RFLP profiles obtained by digesting the rDNA ITS region ofBiomphalariamollusks withDdeI (lanes 1-9) andAluI (lanes 10-14). Lane 1: Biomphalaria straminea (Minas Gerais, Brazil); 2: Biomphalaria intermedia (Minas Gerais, Brazil); 3: Biomphalaria straminea (Corrientes, Argentina); 4: Biomphalaria straminea (Espinillar, Uruguay); 5: Biomphalaria tenagophila (Minas Gerais, Brazil); 6-7: Biomphalaria tenagophila (Corrientes, Argentina); 8: Biomphalaria tenagophila guaibensis (Rio Grande do Sul, Brazil); 9: Biomphalaria occidentalis (Minas Gerais, Brazil); 10: Biomphalaria tenagophila (Minas Gerais, Brazil); 11-12: Biomphalaria tenagophila (Corrientes, Argentina); 13: Biomphalaria tenagophila guaibensis (Rio Grande do Sul, Brazil); 14: Biomphalaria occidentalis (Minas Gerais, Brazil). Values to the left correspond to molecular weights in base pairs (bp).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150920&req=5

Fig1: Biomphalariarestriction profiles: 6% polyacrylamide gel showing the PCR-RFLP profiles obtained by digesting the rDNA ITS region ofBiomphalariamollusks withDdeI (lanes 1-9) andAluI (lanes 10-14). Lane 1: Biomphalaria straminea (Minas Gerais, Brazil); 2: Biomphalaria intermedia (Minas Gerais, Brazil); 3: Biomphalaria straminea (Corrientes, Argentina); 4: Biomphalaria straminea (Espinillar, Uruguay); 5: Biomphalaria tenagophila (Minas Gerais, Brazil); 6-7: Biomphalaria tenagophila (Corrientes, Argentina); 8: Biomphalaria tenagophila guaibensis (Rio Grande do Sul, Brazil); 9: Biomphalaria occidentalis (Minas Gerais, Brazil); 10: Biomphalaria tenagophila (Minas Gerais, Brazil); 11-12: Biomphalaria tenagophila (Corrientes, Argentina); 13: Biomphalaria tenagophila guaibensis (Rio Grande do Sul, Brazil); 14: Biomphalaria occidentalis (Minas Gerais, Brazil). Values to the left correspond to molecular weights in base pairs (bp).
Mentions: Three groups were prominent among the inconclusively identified specimens. 1) Specimens from the provinces of Corrientes, Argentina and Espinillar, Uruguay (four collection points) were morphologically similar to B. straminea, but their restriction profile for the enzyme DdeI differed from that of B. straminea (which had been previously established), here represented by sample from Minas Gerais, Brazil (Figure 1). 2) Specimens from the provinces of Corrientes, Argentina and Espinillar, Uruguay (six collection points) were morphologically similar to B. tenagophila from Brazil, but their restriction profiles for the enzymes DdeI and AluI were similar to those of B. t. guaibensis from Rio Grande do Sul, Brazil (Figure 1). 3) Specimens from 208 collection points in various Brazilian states were morphologically identified as B. peregrina, but although some of these specimens had molecular profiles characteristic of that species, others had the molecular profile of B. oligoza (which had been previously established), here represented by sample from Rio Grande do Sul, Brazil (Figure 2). In an attempt to clarify the identity of these specimens, a portion of the mitochondrial cytochrome c oxidase subunit I (COI) gene was amplified by PCR, and the restriction enzymes ClaI, RsaI, and AluI were used to generate PCR-RFLP profiles. There was no restriction site for the enzymes ClaI and RsaI. For AluI, the specimens morphologically identified as B. peregrina sometimes showed B. peregrina profiles and sometimes showed B. oligoza profiles (data not shown).Figure 1

Bottom Line: 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.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.These studies demonstrate the importance of using more than one technique in taxonomic confirmation and the good preservation of specimens' collection.

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Helmintologia e Malacologia Médica do Centro de Pesquisas René Rachou-Fiocruz, Av. Augusto de Lima 1715, Belo Horizonte, MG 30190-002 Brasil.

ABSTRACT

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.

No MeSH data available.