scholarly journals Differentiation of Hedyotis diffusa and Common Adulterants Based on Chloroplast Genome Sequencing and DNA Barcoding Markers

Plants ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 161
Author(s):  
Mavis Hong-Yu Yik ◽  
Bobby Lim-Ho Kong ◽  
Tin-Yan Siu ◽  
David Tai-Wai Lau ◽  
Hui Cao ◽  
...  
Keyword(s):  
Dna Barcoding ◽  
Bacterial Infections ◽  
Morphological Characteristics ◽  
Intergenic Spacer ◽  
Chloroplast Genomes ◽  
Hedyotis Diffusa ◽  
Species Specific ◽  
Chinese Herbal Tea ◽  
Next Generation Sequencing Ngs ◽  
Chloroplast Genome Sequencing

Chinese herbal tea, also known as Liang Cha or cooling beverage, is popular in South China. It is regarded as a quick-fix remedy to relieve minor health problems. Hedyotis diffusa Willd. (colloquially Baihuasheshecao) is a common ingredient of cooling beverages. H. diffusa is also used to treat cancer and bacterial infections. Owing to the high demand for H. diffusa, two common adulterants, Hedyotis brachypoda (DC.) Sivar and Biju (colloquially Nidingjingcao) and Hedyotis corymbosa (L.) Lam. (colloquially Shuixiancao), are commonly encountered in the market. Owing to the close similarity of their morphological characteristics, it is difficult to differentiate them. Here, we sequenced the complete chloroplast genomes of the three species of Hedyotis using next-generation sequencing (NGS). By comparing the complete chloroplast genomes, we found that they are closely related in the subfamily Rubioideae. We also discovered that there are significant differences in the number and repeating motifs of microsatellites and complex repeats and revealed three divergent hotspots, rps16-trnQ intergenic spacer, ndhD and ycf1. By using these species-specific sequences, we propose new DNA barcoding markers for the authentication of H. diffusa and its two common adulterants.

scholarly journals A Forensic Detection Method for Hallucinogenic Mushrooms via High-Resolution Melting (HRM) Analysis

Genes ◽  
2021 ◽  
Vol 12 (2) ◽  
pp. 199
Author(s):  
Xiaochun Zhang ◽  
Huan Yu ◽  
Qi Yang ◽  
Ziwei Wang ◽  
Ruocheng Xia ◽  
...  
Keyword(s):  
High Resolution ◽  
Dna Barcoding ◽  
High Resolution Melting ◽  
Its Region ◽  
Its Sequencing ◽  
Melting Temperatures ◽  
Hrm Analysis ◽  
National Drug ◽  
Species Specific ◽  
Its1 And Its2

In recent years, trafficking and abuse of hallucinogenic mushrooms have become a serious social problem. It is therefore imperative to identify hallucinogenic mushrooms of the genus Psilocybe for national drug control legislation. An internal transcribed spacer (ITS) is a DNA barcoding tool utilized for species identification. Many methods have been used to discriminate the ITS region, but they are often limited by having a low resolution. In this study, we sought to analyze the ITS and its fragments, ITS1 and ITS2, by using high-resolution melting (HRM) analysis, which is a rapid and sensitive method for evaluating sequence variation within PCR amplicons. The ITS HRM assay was tested for specificity, reproducibility, sensitivity, and the capacity to analyze mixture samples. It was shown that the melting temperatures of the ITS, ITS1, and ITS2 of Psilocybe cubensis were 83.72 ± 0.01, 80.98 ± 0.06, and 83.46 ± 0.08 °C, and for other species, we also obtained species-specific results. Finally, we performed ITS sequencing to validate the presumptive taxonomic identity of our samples, and the sequencing output significantly supported our HRM data. Taken together, these results indicate that the HRM method can quickly distinguish the DNA barcoding of Psilocybe cubensis and other fungi, which can be utilized for drug trafficking cases and forensic science.

Temporal Detection Limits of Remnant Larval Bloodmeals in Nymphal Ixodes scapularis (Say, Ixodida: Ixodidae) Using Two Next-Generation Sequencing DNA Barcoding Assays

2020 ◽  
Author(s):  
Genevieve A M Lumsden ◽  
Evgeny V Zakharov ◽  
Sarah Dolynskyj ◽  
J Scott Weese ◽  
L Robbin Lindsay ◽  
...  
Keyword(s):  
Next Generation Sequencing ◽  
Dna Barcoding ◽  
Ixodes Scapularis ◽  
Life Stage ◽  
Next Generation ◽  
Host Detection ◽  
Host Identification ◽  
Species Specific ◽  
Generation Sequencing ◽  
Assay Detection

Abstract Using next-generation sequencing DNA barcoding, we aimed to determine: 1) if the larval bloodmeal can be detected in Ixodes scapularis nymphs and 2) the post-moult temporal window for detection of the larval bloodmeal. Subsets of 30 nymphs fed on a domestic rabbit (Oryctolagus cuniculus Linnaeus, Lagomorphia: Leporidae) as larvae were reared and frozen at 11 time points post-moult, up to 150 d. Vertebrate DNA was amplified using novel universal (UP) and species-specific primers (SSP) and sequenced for comparison against cytochrome c oxidase subunit I barcodes to infer host identification. Detectable bloodmeals decreased as time since moult increased for both assays. For the SSP assay, detection of bloodmeals decreased from 96.7% (n = 29/30) in day 0 nymphs to 3.3% (n = 1/30) and 6.7% (n = 2/30) at 4- and 5-mo post-moult, respectively. A shorter temporal detection period was achieved with the UP assay, declining from 16.7% (n = 5/30) in day 0 nymphs to 0/30 in 3-d-old nymphs. Bloodmeal detection was nonexistent for the remaining cohorts, with the exception of 1/30 nymphs at 2-mo post-moult. Host detection was significantly more likely using the SSP assay compared to the UP assay in the first three time cohorts (day 0: χ 2 = 39.1, P < 0.005; day 2: χ 2 = 19.2, P < 0.005; day 3: χ 2 = 23.3, P < 0.005). Regardless of the primer set used, the next-generation sequencing DNA barcoding assay was able to detect host DNA from a larval bloodmeal in the nymphal life stage; however, a short window with a high proportion of detection post-moult was achieved.

scholarly journals Bioinformatics Approach to Mitigate Mislabeling in EU Seafood Market and Protect Consumer Health

2021 ◽  
Vol 18 (14) ◽  
pp. 7497
Author(s):  
Gabriella Vindigni ◽  
Alfredo Pulvirenti ◽  
Salvatore Alaimo ◽  
Clara Monaco ◽  
Daniela Spina ◽  
...  
Keyword(s):  
Dna Barcoding ◽  
Value Chain ◽  
Morphological Characteristics ◽  
Molecular Techniques ◽  
Consumer Information ◽  
Fish Market ◽  
Fishery Policy ◽  
Fisheries Policy ◽  
Consumer Interests ◽  
Zoonotic Parasites

Fisheries products are some of the most traded commodities world-wide and the potential for fraud is a serious concern. Fish fraud represents a threat to human health and poses serious concerns due to the consumption of toxins, highly allergenic species, contaminates or zoonotic parasites, which may be present in substituted fish. The substitution of more expensive fish by cheaper species, with similar morphological characteristics but different origins, reflects the need for greater transparency and traceability upon which which the security of the entire seafood value-chain depends. Even though EU regulations have made significant progress in consumer information by stringent labelling requirements, fraud is still widespread. Many molecular techniques such as DNA barcoding provide valuable support to enhance the Common Fisheries Policy (CFP) in the protection of consumer interests by unequivocally detecting any kind of fraud. This paper aims to highlight both the engagement of EU fishery policy and the opportunity offered by new biotechnology instruments to mitigate the growing fraud in the globalized fish market and to enforce the food security system to protect consumers’ health. In this paper, after a presentation of EU rules on fish labeling and a general overview on the current state of the global fish market, we discuss the public health implications and the opportunities offered by several techniques based on genetics, reporting a case study to show the efficacy of the DNA barcoding methodology in assessing fish traceability and identification, comparing different species of the Epinephelus genus, Mottled Grouper (Mycteroperca rubra) and Wreckfish (Polyprion americanus), often improperly sold with the commercial name of “grouper”.

Duplex DNA barcoding allows accurate species determination of morphologically similar limpets (Patella spp.) from non-destructive sampling

2016 ◽  
Vol 97 (7) ◽  
pp. 1479-1482 ◽  
Author(s):  
Thomas J. Ashton ◽  
Meriem Kayoueche-Reeve ◽  
Andrew J. Blight ◽  
Jon Moore ◽  
David M. Paterson
Keyword(s):  
Dna Barcoding ◽  
Species Abundance ◽  
Microscopic Analysis ◽  
Cost Effective ◽  
Field Studies ◽  
Similar Species ◽  
Species Determination ◽  
Destructive Sampling ◽  
Species Specific ◽  
Non Destructive

Accurate discrimination of two morphologically similar species of Patella limpets has been facilitated by using qPCR amplification of species-specific mitochondrial genomic regions. Cost-effective and non-destructive sampling is achieved using a mucus swab and simple sample lysis and dilution to create a PCR template. Results show 100% concurrence with dissection and microscopic analysis, and the technique has been employed successfully in field studies. The use of highly sensitive DNA barcoding techniques such as this hold great potential for improving previously challenging field assessments of species abundance.

scholarly journals Characterization of the Bull's Eye Rot of Apple in Chile

Plant Disease ◽  
2013 ◽  
Vol 97 (4) ◽  
pp. 485-490 ◽  
Author(s):  
Sylvana Soto-Alvear ◽  
Mauricio Lolas ◽  
Inés M. Rosales ◽  
Eduardo R. Chávez ◽  
Bernardo A. Latorre
Keyword(s):  
Morphological Characteristics ◽  
The United States ◽  
Apple Fruit ◽  
Maritime Transportation ◽  
Tubulin Genes ◽  
Internal Transcribed Spacer Sequences ◽  
Apple Production ◽  
Species Specific ◽  
Bull's Eye

Apple fruit in Chile are primarily produced for export to Asia, Europe, and the United States, which typically requires 15 to 40 days of maritime transportation. Therefore, Chilean apple production must fulfill the sanitization requirements imposed by the receiving countries. Under these circumstances, it was important to clarify the etiology of bull's eye rot that can severely affect ‘Cripps Pink’ apple and other late-harvest cultivars in Chile. Based on morphological characteristics and the partial sequence analysis of the internal transcribed spacer sequences and β-tubulin genes, Neofabraea alba was identified as the causal agent of the bull's eye rot of Chilean apple. These results were further corroborated using species-specific primers. The incidence of bull's eye rot varied considerably; for instance, in 2009, 0.0 to 58.7% in 38 Cripps Pink orchards surveyed in the relatively arid and humid apple-growing areas of Chile, respectively. There was no evidence for the presence of N. malicorticis or N. perennans, which are commonly identified as causal agents of bull's eye rot in other apple-producing countries. Altogether, these data suggest that N. alba might represent the predominant and possibly the only cause of bull's-eye rot of Chilean apple.

scholarly journals The Odilorhabdin Antibiotic Biosynthetic Cluster and Acetyltransferase Self-Resistance Locus Are Niche and Species Specific

mBio ◽  
2022 ◽  
Author(s):  
Anne Lanois-Nouri ◽  
Lucile Pantel ◽  
Jun Fu ◽  
Jessica Houard ◽  
Jean-Claude Ogier ◽  
...  
Keyword(s):  
Bacterial Infections ◽  
Multidrug Resistant ◽  
Small Subunit ◽  
Cationic Peptides ◽  
Resistance Locus ◽  
Gram Negative ◽  
Species Specific ◽  
Bacterial Translation

Odilorhabdins (ODLs) constitute a novel antibiotic family with promising properties for treating problematic multidrug-resistant Gram-negative bacterial infections. ODLs are 10-mer linear cationic peptides inhibiting bacterial translation by binding to the small subunit of the ribosome.

scholarly journals First record of the semi-slug Omalonyx unguis (d’Orbigny, 1837) (Gastropoda, Succineidae) in the Misiones Province, Argentina

Check List ◽  
2018 ◽  
Vol 14 (4) ◽  
pp. 705-712
Author(s):  
Leila B. Guzmán ◽  
Enzo N. Serniotti ◽  
Roberto E. Vogler ◽  
Ariel A. Beltramino ◽  
Alejandra Rumi ◽  
...  
Keyword(s):  
Dna Sequences ◽  
Morphological Characteristics ◽  
First Record ◽  
Coi Gene ◽  
Cytochrome Oxidase Subunit ◽  
Specific Identification ◽  
Molluscan Fauna ◽  
Misiones Province ◽  
Species Specific ◽  
Northeastern Argentina

Omalonyx unguis (d’Orbigny, 1837) is a semi-slug inhabiting the Paraná river basin. This species belongs to Succineidae, a family comprising a few representatives in South America. In this work, we provide the first record for the species from Misiones Province, Argentina. Previous records available for Omalonyx in Misiones were identified to the genus level. We examined morphological characteristics of the reproductive system and used DNA sequences from cytochrome oxidase subunit I (COI) gene for species-specific identification. These new distributional data contribute to consolidate the knowledge of the molluscan fauna in northeastern Argentina.

Development of an efficient PCR-based diagnosis protocol for the identification of the pinewood nematode, Bursaphelenchus xylophilus (Nematoda: Aphelenchoididae)

Nematology ◽  
2004 ◽  
Vol 6 (2) ◽  
pp. 279-285 ◽  
Author(s):  
Jae Soon Kang ◽  
Kwang Sik Choi ◽  
Sang Chul Shin ◽  
Il Sung Moon ◽  
Sang Gil Lee ◽  
...  
Keyword(s):  
Intergenic Spacer ◽  
Nematode Species ◽  
Bursaphelenchus Xylophilus ◽  
5S Rrna ◽  
Rrna Gene ◽  
Pinewood Nematode ◽  
Pine Wood ◽  
5S Rrna Gene ◽  
Primer Sets ◽  
Species Specific

Abstract Pine wood wilt disease caused by the pine wood nematode, Bursaphelenchus xylophilus , has been a serious problem in the southern regions of Korea. Efficient diagnosis of B. xylophilus from infected pine wood specimens is critical for the management of this pest. Traditional microscopic examination often results in an erroneous identification because a closely related non-pathogenic species, B. mucronatus, has a great degree of morphological similarity to B. xylophilus. In an attempt to search for reliable molecular markers for the discrimination of these species, we have cloned the 5S rRNA genomic DNA fragments containing both coding and intergenic spacer (IGS) regions from B. xylophilus and B. mucronatus through a homology-probing PCR strategy. Sequence analyses revealed that coding sequences of the 5S rRNA gene from the two species are almost identical (98.3% homology) but that the IGS sequences differ substantially between the species. Based on the IGS sequence differences (69.7% homology), we designed species-specific primer sets and developed a PCR-based diagnosis protocol for the identification and discrimination of the two nematode species on a molecular basis.

DNA Barcoding Revealed Mislabeling and Potential Health Concerns with Roasted Fish Products Sold across China

2019 ◽  
Vol 82 (7) ◽  
pp. 1200-1209 ◽  
Author(s):  
XIONG XIONG ◽  
FANGYING YUAN ◽  
MANHONG HUANG ◽  
LIXIA LU ◽  
XIAOHUI XIONG ◽  
...  
Keyword(s):  
Dna Barcoding ◽  
Fish Species ◽  
Morphological Characteristics ◽  
Economic Loss ◽  
High Price ◽  
Snack Food ◽  
Fish Fillet ◽  
Potential Health ◽  
Fish Products ◽  
Republic Of China

ABSTRACTRoasted fish fillet is a popular snack food in the People's Republic of China, and the market appeal can be reflected by the high price, which exceeds 330 RMB/kg in some cases. However, the labeled fish common name generally cannot be connected with a particular species, and the processing treatments make it a challenging task to identify fish species based on morphological characteristics. This study used full DNA barcoding and mini-DNA barcoding for species identification of 202 specimens sorted from 64 roasted fish fillet products sold across China, which belonged to 32 distinct brands and claimed 16 fish common names on the label. Given the absence of a harmonization in defining these common names in China, scientific literature and the Latin-Chinese dictionary of fish names (http://fishdb.sinica.edu.tw/) were consulted to define at least the family for each common name, and an identified species outside the expected family was determined to be mislabeled. Results highlighted a high mislabeling rate of 75.5%. Even worse, fish species were identified as belonging to multiple families, not only from several products of the same brand under the same common name but also from several specimens sorted from the same product. Moreover, a health issue was highlighted by the identification of toxic Tetraodontidae species from one product. To protect the consumers from economic loss and even health risks, we recommended amendments to existing legislation and the compilation of a list of acceptable market names for fish species in China. Meanwhile, DNA barcoding is a powerful tool in fish forensics, and we recommended the use of this technique to assure the accurate species labeling of fish products.HIGHLIGHTS

scholarly journals Diversity of Aerobic Bacteria Isolated from Oral and Cloacal Cavities from Free-Living Snakes Species in Costa Rica Rainforest

2017 ◽  
Vol 2017 ◽  
pp. 1-9 ◽  
Author(s):  
Allan Artavia-León ◽  
Ariel Romero-Guerrero ◽  
Carolina Sancho-Blanco ◽  
Norman Rojas ◽  
Rodolfo Umaña-Castro
Keyword(s):  
Costa Rica ◽  
Bacterial Infections ◽  
Bacterial Species ◽  
Ribosomal Database Project ◽  
Opportunistic Pathogens ◽  
Bacterial Taxonomy ◽  
Free Living ◽  
16S Rdna Pcr ◽  
Animal Bites ◽  
Species Specific

Costa Rica has a significant number of snakebites per year and bacterial infections are often complications in these animal bites. Hereby, this study aims to identify, characterize, and report the diversity of the bacterial community in the oral and cloacal cavities of venomous and nonvenomous snakes found in wildlife in Costa Rica. The snakes where captured by casual encounter search between August and November of 2014 in the Quebrada González sector, in Braulio Carrillo National Park. A total of 120 swabs, oral and cloacal, were taken from 16 individuals of the Viperidae and Colubridae families. Samples were cultured on four different media at room temperature. Once isolated in pure culture, colonies were identified with the VITEK® 2C platform (bioMérieux). In order to test the identification provided on environmental isolates, molecular analyses were conducted on 27 isolates of different bacterial species. Specific 16S rDNA PCR-mediated amplification for bacterial taxonomy was performed, then sequenced, and compared with sequences of Ribosomal Database Project (RDP). From 90 bacterial isolates, 40 different bacterial species were identified from both oral and cloacal swabs. These results indicate the diversity of opportunistic pathogens present and their potential to generate infections and zoonosis in humans.

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