scholarly journals Comparison between single and multi-locus approaches for specimen identification in Mytilus mussels

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
María Angélica Larraín ◽  
Pía González ◽  
Claudio Pérez ◽  
Cristián Araneda
Keyword(s):  
Species Identification ◽  
Dna Marker ◽  
Morphological Characteristics ◽  
Rflp Markers ◽  
Identification Performance ◽  
As Species ◽  
Pcr Rflp ◽  
Food Quality And Safety ◽  
Genomic Regions ◽  
Specimen Identification

AbstractMytilus mussels have been the object of much research given their sentinel role in coastal ecosystems and significant value as an aquaculture resource appreciated for both, its flavour and nutritional content. Some of the most-studied Mytilus species are M. edulis, M. galloprovincialis, M. chilensis and M. trossulus. As species identification based on morphological characteristics of Mytilus specimens is difficult, molecular markers are often used. Single-locus markers can give conflicting results when used independently; not all markers differentiate among all species, and the markers target genomic regions with different evolutionary histories. We evaluated the concordance between the PCR-RFLP markers most commonly-used for species identification in mussels within the Mytilus genus (Me15-16, ITS, mac-1, 16S rRNA and COI) when used alone (mono-locus approach) or together (multi-locus approach). In this study, multi-locus strategy outperformed the mono-locus methods, clearly identifying all four species and also showed similar specimen identification performance than a 49 SNPs panel. We hope that these findings will contribute to a better understanding of DNA marker-based analysis of Mytilus taxa. These results support the use of a multi-locus approach when studying this important marine resource, including research on food quality and safety, sustainable production and conservation.

scholarly journals Development of SCAR Markers for Species Identification in the Genus Shorea (Dipterocarpaceae)

2010 ◽  
Vol 59 (1-6) ◽  
pp. 249-257 ◽  
Author(s):  
H. S. Nuroniah ◽  
O. Gailing ◽  
R. Finkeldey
Keyword(s):  
Species Identification ◽  
Rflp Markers ◽  
Scar Markers ◽  
Closely Related Species ◽  
Tropical Timber ◽  
Sequence Characterized Amplified Region ◽  
Cpdna Region ◽  
Pcr Rflp ◽  
Primer Sets ◽  
Good Agreement

AbstractThe development of molecular markers unambiguously distinguishing groups at different taxonomic levels has numerous forensic applications. The identification of tropical timber is of particular interest in this context. We describe the development of SCAR (Sequence Characterized Amplified Region) markers for forensic applications taking the example of two closely related species of the tropical tree family Shorea (Dipterocarpaceae). Two AFLP (Amplified Fragment Length Polymorphism) fragments have been described earlier showing strong differentiation between S. leprosula and S. parvifolia. The AFLP markers were isolated from a gel, re-amplified, cloned and sequenced. Primer sets were designed from these sequences and AFLP fragments were converted into SCAR markers. The SCAR markers and PCR-RFLP markers of the chloroplast region trnLF digested with HinfI were used to screen in total 557 samples of S. parvifolia and S. leprosula from nineteen widely separated populations in Indonesia. Complete genetic differentiation between species was observed based on the putatively nuclear SCAR marker and the PCR-RFLP of the cpDNA region. We found a good agreement between leaf morphological variation and species identification based on both marker types and no indication for interspecific hybridization.

Molecular typing of Staphylococcus aureus based on PCR-RFLP of coa gene and RAPD analysis

2011 ◽  
Vol 14 (2) ◽  
pp. 285-286 ◽  
Author(s):  
J. Karakulska ◽  
A. Pobucewicz ◽  
P. Nawrotek ◽  
M. Muszyńska ◽  
A. Furowicz ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Species Identification ◽  
Genetic Relationship ◽  
Molecular Typing ◽  
Rapd Analysis ◽  
Milk Samples ◽  
Rapd Pcr ◽  
Pcr Rflp ◽  
Mastitic Milk ◽  
Coa Gene

Molecular typing ofStaphylococcus aureusbased on PCR-RFLP ofcoagene and RAPD analysisThe aim of this study was molecular identification ofS. aureusstrains isolated from mastitic milk samples and establishing the genetic relationship between strains isolated from cows belonging to the same herd. In all 43 isolated strains thegapgene (930 bp) was amplified, which enabled their affiliation to theStaphylococcusgenus to be established. PCR-RFLP withAluI endonuclease of thegapgene as well asnuc(450 bp) andcoa(1130 bp) gene amplification allowed preciseS. aureusspecies identification. One hundred percent of the genetic relationship between strains was establishedviaRAPD-PCR and coa-typing.

Evidence of the three main clonalToxoplasma gondiilineages from wild mammalian carnivores in the UK

Parasitology ◽  
2013 ◽  
Vol 140 (14) ◽  
pp. 1768-1776 ◽  
Author(s):  
A. BURRELLS ◽  
P. M. BARTLEY ◽  
I. A. ZIMMER ◽  
S. ROY ◽  
A. C. KITCHENER ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Rflp Markers ◽  
Type I ◽  
Type Ii ◽  
Red Foxes ◽  
Mammal Species ◽  
Tissue Samples ◽  
Pcr Rflp ◽  
Few Data ◽  
The Uk

SUMMARYToxoplasma gondiiis a zoonotic pathogen defined by three main clonal lineages (types I, II, III), of which type II is most common in Europe. Very few data exist on the prevalence and genotypes ofT. gondiiin the UK. Wildlife can act as sentinel species forT. gondiigenotypes present in the environment, which may subsequently be transmitted to livestock and humans. DNA was extracted from tissue samples of wild British carnivores, including 99 ferrets, 83 red foxes, 70 polecats, 65 mink, 64 badgers and 9 stoats. Parasite DNA was detected using a nested ITS1 PCR specific forT. gondii, PCR positive samples were subsequently genotyped using five PCR–RFLP markers.Toxoplasma gondiiDNA was detected within all these mammal species and prevalence varied from 6·0 to 44·4% depending on the host. PCR–RFLP genotyping identified type II as the predominant lineage, but type III and type I alleles were also identified. No atypical or mixed genotypes were identified within these animals. This study demonstrates the presence of alleles for all three clonal lineages with potential for transmission to cats and livestock. This is the first DNA-based study ofT. gondiiprevalence and genotypes across a broad range of wild British carnivores.

scholarly journals Phylogenetic Relationships of Local Durian Species based on Morphological Characteristics and PCR-RFLP Analysis of the Ribosomal Internal Transcribed Spacer (ITS) DNA

2016 ◽  
Vol 8 (3) ◽  
pp. 362
Author(s):  
Fidia Fibriana ◽  
Lutfia Nur Hadiyanti
Keyword(s):  
Phylogenetic Relationships ◽  
Internal Transcribed Spacer ◽  
Morphological Characteristics ◽  
Biology Education ◽  
Rflp Analysis ◽  
Restriction Pattern ◽  
Internal Transcribed Spacers ◽  
Central Java ◽  
Pcr Rflp ◽  
Pcr Products

<p>In this study, twenty local durian accessions obtained from Central Java in situ collection were characterized using the morphological characteristics and the restriction patterns which generated from the region spanning the internal transcribed spacers ITS LEU and ITS 4. Morphological characteristics of durian leaf, stem, tree, and fruit showed variations for the different accessions, whereas polymerase chain reaction (PCR) products of ribosomal DNA region showed a low length of variation. The size of the PCR products and the restriction analyses with the restriction endonucleases Bsp1431yielded a restriction pattern for each accessions. The results of this study can be utilized by local durian farmers as a preliminary reference for durian propagation. The data obtained need to be supported by further research using the other molecular markers to obtain more accurate data. The clear identity of durian species can help the management of propagation systems by farmers to get superior local durian.</p><p><strong>How to Cite</strong></p><p>Fibriana, F., &amp; Hadiyanti, L. N. (2016). Phylogenetic Relationships of Local Durian Species based on Morphological Characteristics and PCR-RFLP Analysis of the Ribosomal Internal Transcribed Spacer (ITS) DNA. <em>Biosaintifika: Journal of Biology &amp; Biology Education</em>, 8(3), 362-370. </p>

scholarly journals Species Identification of Five Penaeid Shrimps Using PCR-RFLP and SSCP Analyses of 16S Ribosomal DNA

BMB Reports ◽  
2005 ◽  
Vol 38 (4) ◽  
pp. 491-499 ◽  
Author(s):  
Bavornlak Khamnamtong ◽  
Sirawut Klinbunga ◽  
Piamsak Menasveta
Keyword(s):  
Species Identification ◽  
Ribosomal Dna ◽  
16S Ribosomal Dna ◽  
Pcr Rflp ◽  
Penaeid Shrimps

scholarly journals A genomic perspective on the taxonomy of the subtribe Carcharodina (Lepidoptera: Hesperiidae: Carcharodini)

Zootaxa ◽  
2020 ◽  
Vol 4748 (1) ◽  
pp. 182-194 ◽  
Author(s):  
JING ZHANG ◽  
ERNST BROCKMANN ◽  
QIAN CONG ◽  
JINHUI SHEN ◽  
NICK V. GRISHIN
Keyword(s):  
Dna Sequences ◽  
Type Species ◽  
Phylogenetic Trees ◽  
Type Specimens ◽  
African Species ◽  
Protein Coding ◽  
Coding Regions ◽  
As Species ◽  
Close Relatives ◽  
Genomic Regions

We obtained whole genome shotgun sequences and phylogenetically analyzed protein-coding regions of representative skipper butterflies from the genus Carcharodus Hübner, [1819] and its close relatives. Type species of all available genus-group names were sequenced. We find that species attributed to four exclusively Old World genera (Spialia Swinhoe, 1912, Gomalia Moore, 1879, Carcharodus Hübner, [1819] and Muschampia Tutt, 1906) form a monophyletic group that we call a subtribe Carcharodina Verity, 1940. In the phylogenetic trees built from various genomic regions, these species form 7 (not 4) groups that we treat as genera. We find that Muschampia Tutt, 1906 is not monophyletic, and the 5th group is formed by currently monotypic genus Favria Tutt, 1906 new status (type species Hesperia cribrellum Eversmann, 1841), which is sister to Gomalia. The 6th and 7th groups are composed of mostly African species presently placed in Spialia. These groups do not have names and are described here as Ernsta Grishin, gen. n. (type species Pyrgus colotes Druce, 1875) and Agyllia Grishin, gen. n. (type species Pyrgus agylla Trimen, 1889). Two subgroups are recognized in Ernsta: the nominal subgenus and a new one: Delaga Grishin, subgen. n. (type species Pyrgus delagoae Trimen, 1898). Next, we observe that Carcharodus is not monophyletic, and species formerly placed in subgenera Reverdinus Ragusa, 1919 and Lavatheria Verity, 1940 are here transferred to Muschampia. Furthermore, due to differences in male genitalia or DNA sequences, we reinstate Gomalia albofasciata Moore, 1879 and Gomalia jeanneli (Picard, 1949) as species, not subspecies or synonyms of Gomalia elma (Trimen, 1862), and Spialia bifida (Higgins, 1924) as a species, not subspecies of Spialia zebra (Butler, 1888). Sequencing of the type specimens reveals 2.2-3.2% difference in COI barcodes, the evidence that combined with wing pattern differences suggests a new status of a species for Spialia lugens (Staudinger, 1886) and Spialia carnea (Reverdin, 1927), formerly subspecies of Spialia orbifer (Hübner, [1823]). 

scholarly journals A PCR Method That Can Be Further Developed into PCR-RFLP Assay for Eight Animal Species Identification

2018 ◽  
Vol 2018 ◽  
pp. 1-6 ◽  
Author(s):  
Feng Guan ◽  
Yu-Ting Jin ◽  
Jin Zhao ◽  
Ai-Chun Xu ◽  
Yuan-Yuan Luo
Keyword(s):  
Species Identification ◽  
Animal Species ◽  
Limit Of Detection ◽  
Cost Effective ◽  
Universal Primers ◽  
Universal Primer ◽  
Routine Identification ◽  
Pcr Rflp ◽  
Fast Processing ◽  
Rflp Assay

There are many PCR-based methods for animal species identification; however, their detection numbers are limited or could not identify unknown species. We set out to solve this problem by developing a universal primer PCR assay for simultaneous identification of eight animal species, including goat, sheep, deer, buffalo, cattle, yak, pig, and camel. In this assay, the variable lengths of mitochondrial DNA were amplified using a pair of universal primers. PCR amplifications yielded 760 bp, 737 bp, 537 bp, 486 bp, 481 bp, 464 bp, 429 bp, and 359 bp length fragments for goat, sheep, deer, buffalo, cattle, yak, pig, and camel, respectively. This primer pair had no cross-reaction with other common domestic animals and fish. The limit of detection varied from 0.01 to 0.05 ng of genomic DNA for eight animal species in a 20 µl PCR mixture. Each PCR product could be further digested into fragments with variable sizes and qualitative analysis by SspI restriction enzyme. This developed PCR-RFLP assay was sufficient to distinguish all targeted species. Compared with the previous published related methods, this approach is simple, with high throughput, fast processing rates, and more cost-effective for routine identification of meat in foodstuffs.

Molecular and biological characterization ofToxoplasma gondiiisolates from free-range chickens from Guyana, South America, identified several unique and common parasite genotypes

Parasitology ◽  
2007 ◽  
Vol 134 (11) ◽  
pp. 1559-1565 ◽  
Author(s):  
J. P. DUBEY ◽  
L. APPLEWHAITE ◽  
N. SUNDAR ◽  
G. V. VELMURUGAN ◽  
L. A. BANDINI ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
South America ◽  
Rflp Markers ◽  
Free Range ◽  
Clonal Type ◽  
Modified Agglutination Test ◽  
Pcr Rflp ◽  
Free Ranging ◽  
Common Parasite

SUMMARYThe prevalence ofToxoplasma gondiiin free-ranging chickens (Gallus domesticus) is a good indicator of the prevalence ofT. gondiioocysts in the soil because chickens feed from the ground. The prevalence ofT. gondiiin 76 free-range chickens from Guyana, South America was determined. Antibodies toT. gondiiwere assayed by the modified agglutination test (MAT), and found in 50 (65·8%) of 76 chickens with titres of 1:5 in four, 1:10 in one, 1:20 in five, 1:40 in seven, 1:80 in six, 1:160 in eight, 1:320 in four, 1:640 or higher in 15. Hearts and brains of 26 chickens with titres of <1:5 were pooled in 5 batches and bioassayed in mice. Hearts and brains of 50 chickens with titres of 1:5 or higher were bioassayed in mice.Toxoplasma gondiiwas isolated by bioassay in mice from 35 chickens with MAT titres of 1:20 or higher. All mice inoculated with tissues of 30 infected chickens remained asymptomatic.Toxoplasma gondiiisolates from 35 chickens were genotyped using 11 PCR-RFLP markers including SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, a new SAG2, and Apico. A total of 9 genotypes were identified, with 5 genotypes (nos 1, 4, 5, 6 and 7) unique to Guyana, 2 genotypes (nos 2 and 3) previously identified in chickens from Brazil, 1 genotype (no. 8) previously identified in chickens from Brazil, Costa Rica and Nicaragua, and 1 genotype (no. 9) belonging to the clonal type III lineage that exists globally. Infection with 2 genotypes was found from 1 chicken. This is the first report of genetic characterization ofT. gondiiisolates from any host from Guyana.

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