Rapid and zero-cost DNA extraction from soft-bodied insects for routine PCR-based applications

PLoS ONE, Journal Year: 2022, Volume and Issue: 17(7), P. e0271312 - e0271312

Published: July 15, 2022

Nucleic acid extraction is the first and foremost step in molecular biology studies. Extraction of DNA from small, soft-bodied insects often time-consuming costly. A fast, easy, cost-effective method with greater yield purity would aid rapid diagnostics, screening large populations, other routine PCR-based applications. The present study evaluated standardized a zero-cost small for Five methods viz. sterile distilled water (SDW), 1X phosphate-buffered saline (PBS, pH 7.4), 1.4 M sodium chloride (NaCl), 20 mM ethylenediaminetetraacetic (EDTA, 8.0), elution blotted nitrocellulose membrane (NCM) were compared standard CTAB buffer DNeasy® Blood Tissue Kit methods. average yield, purity, storage stability, time, cost assessed all compared. by simply crushing SDW was ideal terms storability, performing applications including detection pathogens vector species. could be accomplished 2.5 min only zero-reagent cost. useful diagnostics populations insects.

Language: Английский

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Taxonomic identification accuracy from BOLD and GenBank databases using over a thousand insect DNA barcodes from Colombia

PLoS ONE, Journal Year: 2023, Volume and Issue: 18(4), P. e0277379 - e0277379

Published: April 24, 2023

Recent declines of insect populations at high rates have resulted in the need to develop a quick method determine their diversity and process massive data for identification species highly diverse groups. A short sequence DNA from COI is widely used by comparing it against sequences known species. Repositories are available online with tools that facilitate matching interest individual. However, performance these can differ. Here we aim assess accuracy taxonomic categories two repositories, BOLD Systems GenBank. This was done matches between taxonomist suggested platforms. We 1,160 representing eight orders insects Colombia. After comparison, reanalyzed results representative subset subfamily Scarabaeinae (Coleoptera). Overall, systems outperformed GenBank, both engines differed other (species, genus family). Higher accurate were obtained family levels. The higher order Coleoptera level, Lepidoptera level. Other performed similarly repositories. Moreover, showed correctly identified only when match percentage above 93.4% total 85% samples assigned category. These accentuate great potential place accurately into respective based on barcodes highlight reliability absence large reference database country.

Language: Английский

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A Real-Time PCR Assay for Detecting Codling Moth Cydia pomonella on Material Intercepted at U.S. Ports of Entry—A Valuable Tool for Specimen Identification

International Journal of Molecular Sciences, Journal Year: 2025, Volume and Issue: 26(2), P. 707 - 707

Published: Jan. 15, 2025

Codling moth Cydia pomonella is well established nearly everywhere apples are grown. Due to this almost global distribution, larvae often intercepted at U.S. ports of entry where immature specimens cannot be identified accurately species leading unnecessary quarantine actions. To assist with identifying C. from port inspections, we developed a probe-based real-time PCR assay amplify the internal transcribed spacer (ITS) region 2 pomonella. The was tested for inclusivity using 110 six continents. Analytical specificity examined by testing related entry, as non-targets same geographic distribution and host here all individuals correctly produced appropriately negative results non-target species. These ensure that provides rapid accurate tool unambiguously among material entry. Since not actionable, ability identify life stages conclusively will save time, effort, money while also directing identification efforts towards current concern.

Language: Английский

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Evaluation of Environmental DNA as a Surveillance Tool for Invasive House Mice (Mus musculus)

Environmental DNA, Journal Year: 2025, Volume and Issue: 7(1)

Published: Jan. 1, 2025

ABSTRACT Increasing the success of invasive species management depends on development, testing, and deployment new tools. Environmental DNA (eDNA) is an effective tool for monitoring that can help identify presence/absence, geographical boundaries invasion, risk pathways, population connectivity. In particular, understanding sensitivity eDNA detection rates to target density allows calibration sampling rates. this study, we take a lab‐validated assay Mus musculus (house mouse) test its at different populations densities wild‐caught, free‐ranging M. in controlled laboratory outdoor mesocosm. The goal was understand both accumulation after introduced persistence accumulated signal environment animals were removed. We found detectable within 1 h single mouse being months but largely undetectable 4 days suggest strategies post‐eradication highlight other uses assay, which are important management.

Language: Английский

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Tracking Biosecurity Through the Diversity and Network Structure of International Trade

Diversity, Journal Year: 2025, Volume and Issue: 17(3), P. 213 - 213

Published: March 14, 2025

Effective and evidence-based biosecurity measures are essential to prevent trade disruption, protect industries contain the chains of biological invasions. There increasing demands for analysts use quantitative data monitor this system, with goals early detection forecasting. However, standard approaches often struggle incomplete complex nature data, which tends include non-normality, temporal spatial autocorrelation, limited observations. In study, a time series open access import spanning three years was used generate diversity indices network topology, alongside detailed analyses pathways interception records harmful organisms, revealing their dynamic patterns across different routes. Patterns annual seasonality were evident board. A combination Inverse Simpson’s Linkage density optimised monitoring power interceptions taxa. Traditional correlations total number remained intractable, but machine learning tools demonstrated predictive forecast these patterns. Combined, methods provide novel approach in plant animal international borders. These indicators complement more conventional economic metrics, giving actionable insights into complexity status.

Language: Английский

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Turfgrass Disease Diagnosis: Past, Present, and Future

Plants, Journal Year: 2020, Volume and Issue: 9(11), P. 1544 - 1544

Published: Nov. 11, 2020

Turfgrass is a multibillion-dollar industry severely affected by plant pathogens including fungi, bacteria, viruses, and nematodes. Many of the diseases in turfgrass have similar signs symptoms, making it difficult to diagnose specific problem pathogen. Incorrect diagnosis leads delay treatment excessive use chemicals. To effectively control these diseases, important rapid accurate detection systems early stages infection that harbor relatively low pathogen populations. There are many methods for diagnosing on turfgrass. Traditional include morphology, microscopy identification. These been followed nucleic acid onsite techniques. allow diagnosis, some even within field without much expertise. several great potential, such as high-throughput sequencing remote sensing. Utilization techniques disease allows faster reduction damage cost control. Understanding each can researchers select which method best suited their interest. The objective this article provide an overview diagnostics efforts used highlight prospects detection.

Language: Английский

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DNA barcoding of insects from India: Current status and future perspectives

Molecular Biology Reports, Journal Year: 2022, Volume and Issue: 49(11), P. 10617 - 10626

Published: June 18, 2022

Language: Английский

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Monitoring and Surveillance of Forest Insects

Springer eBooks, Journal Year: 2023, Volume and Issue: unknown, P. 669 - 705

Published: Jan. 1, 2023

Language: Английский

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Phylogeographic Reconstruction to Trace the Source Population of Asian Giant Hornet Caught in Nanaimo in Canada and Blaine in the USA

Life, Journal Year: 2024, Volume and Issue: 14(3), P. 283 - 283

Published: Feb. 20, 2024

The Asian giant hornet, Vespa mandarinia, is an invasive species that could potentially destroy the local honeybee industry in North America. It has been observed to nest coastal regions of British Columbia Canada and Washington State USA. What source population immigrant hornets? identification can shed light not only on route immigration but also similarity between native habitat potential new Pacific Northwest. We analyzed mitochondrial COX1 sequences specimens sampled from multiple populations China, Republic Korea, Japan, Russian Far East. V. mandarinia exhibits phylogeographic patterns, forming monophyletic clades for 16 six two Japan. Nanaimo, Columbia, are identical sequence Blaine, State, clustered with those Korea one Korea. Our geophylogeny, which allows visualization genetic variation over time space, provides evolutionary insights evolution speciation three closely related vespine (V. tropica, soror, mandarinia), events associated expansion distribution north.

Language: Английский

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Capacity of United States federal government and its partners to rapidly and accurately report the identity (taxonomy) of non-native organisms intercepted in early detection programs

Biological Invasions, Journal Year: 2019, Volume and Issue: 22(1), P. 101 - 127

Published: Dec. 31, 2019

Abstract The early detection of and rapid response to invasive species (EDRR) depends on accurate identification non-native species. 2016–2018 National Invasive Species Council Management Plan called for an assessment US government (federal) capacity report the identity organisms intercepted through programs. This paper serves as that action item. Here we summarize survey-based findings make recommendations improving federal government’s identify authoritatively in a timely manner. We conclude with improve within context EDRR by increasing coordination, maintaining taxonomic expertise, creating tools clearinghouse, developing using standards naming protocols, expanding content DNA Barcode libraries, ensuring long-term sustainability biological collections, engaging empowering citizens citizen science groups.

Language: Английский

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