Abstract:Among oysters, species of Crassostrea (Sacco, 1897) are the most attractive to aquaculture. In Brazil, the genus is represented by C. rhizophorae (Guilding, 1828) and C. brasiliana (Lamarck, 1819). Because the maturation and breeding technology is not well developed for these species, aquaculturists need a reliable method to decide the correct time to place spat collectors in the field, and to identify both species, which are morphologically similar. In this study a specific Multiplex PCR protocol was develope… Show more
“…Oysters used in this research were identified initially by morphological characteristics and by the position they occupy in the water column, as described by Castilho-Westphal (2012). This preliminary identification was confirmed by sampling (n = 263) and further application of molecular protocols developed by Ludwig et al (2011). Of these, 59% belonged to the C. brasiliana species, 38% belonged to C. rhizophorae, and 3% were classified as Crassostrea sp.…”
Section: Species Identificationmentioning
confidence: 64%
“…This preliminary identification was confirmed by sampling ( n = 263) and further application of molecular protocols developed by Ludwig et al . (). Of these, 59% belonged to the C. brasiliana s pecies, 38% belonged to C. rhizophorae , and 3% were classified as Crassostrea sp.…”
Section: Resultsmentioning
confidence: 97%
“…The adductor muscle fragments were preserved in a dimethyl sulphoxide (DMSO)-ethylenediaminetetraacetic acid (EDTA) buffer to identify the species via the polymerase chain reaction (PCR)-restriction fragment length polymorphisms (RFLP) protocol developed by Ludwig et al (2011). Of the animals collected for gonadal histology, 58% were used to confirm their species identity.…”
This study aimed to describe the gonadal histology and the reproductive cycle of Crassostrea brasiliana in the mangroves of Guaratuba Bay in southern Brazil. Adults were collected monthly from January 2010 to April 2011 from three sampling sites in intertidal oyster beds. The animals were evaluated using biometric and histological analyses of the gonads. The gonadal tissue samples were processed according to the standard histological procedures, and permanent slides were prepared using Harris' haematoxylin and eosin. The oysters were identified at the species level using a molecular protocol. Females (69%) predominated over males (26%), with 4% indeterminate and 1% hermaphroditic. Mature females were more prevalent in February, March and December 2010 and in March 2011. Mature males were more prevalent in February and April 2010 and in March 2011. The presence of hermaphroditic individuals was sporadic, and oysters in immature stages or sexual repose were observed in only a few collections between the months of May and October 2010. The reproduction of C. brasiliana in Guaratuba Bay occurs intermittently, but with greater intensity during the summer, with a larger number of females produced.
“…Oysters used in this research were identified initially by morphological characteristics and by the position they occupy in the water column, as described by Castilho-Westphal (2012). This preliminary identification was confirmed by sampling (n = 263) and further application of molecular protocols developed by Ludwig et al (2011). Of these, 59% belonged to the C. brasiliana species, 38% belonged to C. rhizophorae, and 3% were classified as Crassostrea sp.…”
Section: Species Identificationmentioning
confidence: 64%
“…This preliminary identification was confirmed by sampling ( n = 263) and further application of molecular protocols developed by Ludwig et al . (). Of these, 59% belonged to the C. brasiliana s pecies, 38% belonged to C. rhizophorae , and 3% were classified as Crassostrea sp.…”
Section: Resultsmentioning
confidence: 97%
“…The adductor muscle fragments were preserved in a dimethyl sulphoxide (DMSO)-ethylenediaminetetraacetic acid (EDTA) buffer to identify the species via the polymerase chain reaction (PCR)-restriction fragment length polymorphisms (RFLP) protocol developed by Ludwig et al (2011). Of the animals collected for gonadal histology, 58% were used to confirm their species identity.…”
This study aimed to describe the gonadal histology and the reproductive cycle of Crassostrea brasiliana in the mangroves of Guaratuba Bay in southern Brazil. Adults were collected monthly from January 2010 to April 2011 from three sampling sites in intertidal oyster beds. The animals were evaluated using biometric and histological analyses of the gonads. The gonadal tissue samples were processed according to the standard histological procedures, and permanent slides were prepared using Harris' haematoxylin and eosin. The oysters were identified at the species level using a molecular protocol. Females (69%) predominated over males (26%), with 4% indeterminate and 1% hermaphroditic. Mature females were more prevalent in February, March and December 2010 and in March 2011. Mature males were more prevalent in February and April 2010 and in March 2011. The presence of hermaphroditic individuals was sporadic, and oysters in immature stages or sexual repose were observed in only a few collections between the months of May and October 2010. The reproduction of C. brasiliana in Guaratuba Bay occurs intermittently, but with greater intensity during the summer, with a larger number of females produced.
“…Multiplex PCR techniques have been developed specifically for the identification of oysters of the genus Crassostrea (LUDWIG et al, 2011;MELO et al, 2013), with the objective of optimizing the taxonomic identification of these organisms, providing an important tool for aquaculture and fishery operations. In addition to this approach, a fragment of approximately 650 base pairs of the 5' extremity of the mitochondrial Cytochrome oxidase subunit I (COI) gene has been adopted as a universal identification system for animal species, including the oysters of the Atlantic Ocean.…”
Oysters are found along the whole coast of Brazil. The phenotypes of the species vary considerably according to the characteristics of the habitat. The present study investigated the existence of different oyster species of the genus Crassostrea on the coast of Maranhão, using the Multiplex PCR technique and DNA Barcoding. The results of the Multiplex PCR revealed two distinct bands characteristic of the species C. gasar and C. rhizophorae in a total of 135 samples analyzed. The sequencing of the COI gene of 98 samples produced a 695 bp fragment and 15 haplotypes for C. gasar and 640 bp and eight haplotypes for C. rhizophorae. The haplotype tree divided the two species clearly into different clades with 100% bootstrap support. Intraspecific genetic divergence was 0.2% in both species, while interspecific divergence was 23.6%. The similarity between the sequences generated and those available in BoldSystems ranged from 97.01% to 98.37% for C. rhizophorae and from 97.55% to 99.84% for both C. gasar and C. brasiliana, reinforcing the taxonomic problems in this group, which supports the synonymization of these species. The DNA barcoding permitted the reliable identification of the samples and confirmed the existence of two species of oyster in the study area.
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