scholarly journals Discrimination between Asian populations of the parasitoid wasp Ganaspis cf. brasiliensis using a simple MALDI-TOF MS-based method for use with insects

2019 ◽  
Vol 4 (1) ◽  
Author(s):  
Michael A Reeve ◽  
M Lukas Seehausen
Keyword(s):  
Reference Sample ◽  
Principal Component ◽  
Parasitoid Wasp ◽  
Test Sample ◽  
Invasive Pest ◽  
Drosophila Suzukii ◽  
Maldi Tof Ms ◽  
Chinese Populations ◽  
Maldi Tof ◽  
Tof Ms

Abstract The fruit fly Drosophila suzukii has recently become an invasive pest insect of significant economic impact in Europe and the USA. In contrast to other Drosophila species, D. suzukii is able to infest intact fruit by means of a saw-like ovipositor, which allows females to deposit eggs beneath the skin of the fruit. Classical biological control using the parasitoid wasp Ganaspis cf. brasiliensis is currently being researched as an environmentally sustainable option for the control of D. suzukii. In particular, the host specificity of this parasitoid has been assessed for populations from different regions in China and Japan. In order to study the relationship between the differences in specificity and molecular variations, we have adapted a matrix-assisted laser-desorption and ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based method, originally developed for use with plant material, to discriminate between example populations of G. cf. brasiliensis. We have employed a combination of principal component analysis and blind-tested comparison between reference sample MALDI-TOF MS spectra and test sample spectra to discriminate, on the basis of the acid-soluble insect protein spectra generated, between four populations of G. cf. brasiliensis (originally collected from Tokyo and Hasuike in Japan and Dali and Ximing in China). MALDI-TOF MS analysis is able to discriminate with 100% accuracy between populations G. cf. brasiliensis. The Chinese populations were observed to be similar, but the Tokyo population is slightly different and the Hasuike population is significantly different from the other populations. The Tokyo population appears more closely related to the Chinese populations than the Hasuike population, even though both originate from Japan.

scholarly journals Novel Method for Reliable Identification of Siccibacter and Franconibacter Strains: from “Pseudo-Cronobacter” to New Enterobacteriaceae Genera

2017 ◽  
Vol 83 (13) ◽  
Author(s):  
Barbora Svobodová ◽  
Jiří Vlach ◽  
Petra Junková ◽  
Ludmila Karamonová ◽  
Martina Blažková ◽  
...  
Keyword(s):  
Intact Cell ◽  
Foodborne Pathogen ◽  
Principal Component ◽  
Powdered Infant Formula ◽  
Content Type ◽  
Reliable Identification ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Maldi Biotyper ◽  
Tof Ms

ABSTRACT In the last decade, strains of the genera Franconibacter and Siccibacter have been misclassified as first Enterobacter and later Cronobacter. Because Cronobacter is a serious foodborne pathogen that affects premature neonates and elderly individuals, such misidentification may not only falsify epidemiological statistics but also lead to tests of powdered infant formula or other foods giving false results. Currently, the main ways of identifying Franconibacter and Siccibacter strains are by biochemical testing or by sequencing of the fusA gene as part of Cronobacter multilocus sequence typing (MLST), but in relation to these strains the former is generally highly difficult and unreliable while the latter remains expensive. To address this, we developed a fast, simple, and most importantly, reliable method for Franconibacter and Siccibacter identification based on intact-cell matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS). Our method integrates the following steps: data preprocessing using mMass software; principal-component analysis (PCA) for the selection of mass spectrum fingerprints of Franconibacter and Siccibacter strains; optimization of the Biotyper database settings for the creation of main spectrum projections (MSPs). This methodology enabled us to create an in-house MALDI MS database that extends the current MALDI Biotyper database by including Franconibacter and Siccibacter strains. Finally, we verified our approach using seven previously unclassified strains, all of which were correctly identified, thereby validating our method. IMPORTANCE We show that the majority of methods currently used for the identification of Franconibacter and Siccibacter bacteria are not able to properly distinguish these strains from those of Cronobacter. While sequencing of the fusA gene as part of Cronobacter MLST remains the most reliable such method, it is highly expensive and time-consuming. Here, we demonstrate a cost-effective and reliable alternative that correctly distinguishes between Franconibacter, Siccibacter, and Cronobacter bacteria and identifies Franconibacter and Siccibacter at the species level. Using intact-cell MALDI-TOF MS, we extend the current MALDI Biotyper database with 11 Franconibacter and Siccibacter MSPs. In addition, the use of our approach is likely to lead to a more reliable identification scheme for Franconibacter and Siccibacter strains and, consequently, a more trustworthy epidemiological picture of their involvement in disease.

Principal component analysis of MALDI-TOF MS of whole-cell foodborne pathogenic bacteria

2020 ◽  
Vol 592 ◽  
pp. 113582 ◽  
Author(s):  
Wenjing Yan ◽  
Jing Qian ◽  
Yongjie Ge ◽  
Keping Ye ◽  
Cunshan Zhou ◽  
...  
Keyword(s):  
Principal Component Analysis ◽  
Pathogenic Bacteria ◽  
Principal Component ◽  
Component Analysis ◽  
Whole Cell ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Foodborne Pathogenic Bacteria ◽  
Tof Ms

scholarly journals MALDI-TOF MS-based analysis of dried seed proteins immobilized on filter paper

2019 ◽  
Vol 4 (1) ◽  
Author(s):  
Michael A Reeve ◽  
Kathryn M Pollard
Keyword(s):  
Biological Control ◽  
Filter Paper ◽  
Time Course ◽  
Biological Control Agent ◽  
Seed Proteins ◽  
Principal Component ◽  
Impatiens Glandulifera ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Tof Ms

Abstract Matrix-assisted laser-desorption and ionization time-of-flight mass spectroscopy (MALDI-TOF MS) is commonly used for the characterization of protein-containing biological samples. For this, we have previously developed sample-preparation methods that can be used for discrimination between Impatiens species and also between regional biotypes of Himalayan balsam (Impatiens glandulifera), initially using leaf samples and, more recently, using seed material. In the current article, we have developed a further MALDI-TOF MS-based method that can be used with seeds that uses only simple equipment and minimally hazardous reagents prior to storing and/or shipping dried seed proteins immobilized on filter paper for MALDI-TOF MS analysis. We have investigated I. glandulifera regional-biotype seeds originating from four different sites within the UK for which the parent plants differ in their susceptibility to the biological control agent Puccinia komarovii var. glanduliferae. Using a combination of time-course comparisons and principal-component analysis, we have demonstrated good MALDI-TOF MS spectral conservation, even after storage for 1 month at 35°C, of dried seed-protein samples immobilized on filter paper. This method may provide a further useful tool for the matching of biological control agents optimally to susceptible (regional) target-plant biotypes, and for seed characterization and/or identification in general.

Collagen-based proteinaceous binder-pigment interaction study under UV ageing conditions by MALDI-TOF-MS and principal component analysis

2012 ◽  
Vol 47 (3) ◽  
pp. 322-330 ◽  
Author(s):  
Julia Romero-Pastor ◽  
Natalia Navas ◽  
Stepanka Kuckova ◽  
Alejandro Rodríguez-Navarro ◽  
Carolina Cardell
Keyword(s):  
Principal Component Analysis ◽  
Principal Component ◽  
Component Analysis ◽  
Interaction Study ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Uv Ageing ◽  
Proteinaceous Binder ◽  
Tof Ms

scholarly journals Discrimination of Bacillus cereus Group Members by MALDI-TOF Mass Spectrometry

2021 ◽  
Vol 9 (6) ◽  
pp. 1202
Author(s):  
Viviana Manzulli ◽  
Valeria Rondinone ◽  
Alessandro Buchicchio ◽  
Luigina Serrecchia ◽  
Dora Cipolletta ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Bacillus Cereus ◽  
Mass Spectra ◽  
Critical Time ◽  
Principal Component ◽  
Etiologic Agent ◽  
Bacillus Cereus Group ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Tof Ms

Matrix-Assisted Laser Desorption/Ionization Time Of Flight Mass Spectrometry (MALDI-TOF MS) technology is currently increasingly used in diagnostic laboratories as a cost effective, rapid and reliable routine technique for the identification and typing of microorganisms. In this study, we used MALDI-TOF MS to analyze a collection of 160 strains belonging to the Bacillus cereus group (57 B. anthracis, 49 B. cereus, 1 B. mycoides, 18 B. wiedmannii, 27 B. thuringiensis, 7 B. toyonensis and 1 B. weihenstephanensis) and to detect specific biomarkers which would allow an unequivocal identification. The Main Spectra Profiles (MSPs) were added to an in-house reference library, expanding the current commercial library which does not include B. toyonensis and B. wiedmannii mass spectra. The obtained mass spectra were statistically compared by Principal Component Analysis (PCA) that revealed seven different clusters. Moreover, for the identification purpose, were generated dedicate algorithms for a rapid and automatic detection of characteristic ion peaks after the mass spectra acquisition. The presence of specific biomarkers can be used to differentiate strains within the B. cereus group and to make a reliable identification of Bacillus anthracis, etiologic agent of anthrax, which is the most pathogenic and feared bacterium of the group. This could offer a critical time advantage for the diagnosis and for the clinical management of human anthrax even in case of bioterror attacks.

scholarly journals Principal component analysis of MALDI TOF MS mass spectra separates M. abscessus (sensu stricto) from M. massiliense isolates

2016 ◽  
Vol 16 (1) ◽  
Author(s):  
Jan Kehrmann ◽  
Sarah Wessel ◽  
Roshni Murali ◽  
Annegret Hampel ◽  
Franz-Christoph Bange ◽  
...  
Keyword(s):  
Principal Component Analysis ◽  
Mass Spectra ◽  
Principal Component ◽  
Component Analysis ◽  
Sensu Stricto ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Tof Ms

scholarly journals Hymenopteran Parasitoids of Hard Ticks in Western Africa and the Russian Far East

2020 ◽  
Vol 8 (12) ◽  
pp. 1992
Author(s):  
Mapenda Gaye ◽  
Nadia Amanzougaghene ◽  
Younes Laidoudi ◽  
El Hadji Amadou Niang ◽  
Zuzana Sekeyová ◽  
...  
Keyword(s):  
Russian Far East ◽  
Parasitoid Wasp ◽  
Morphological Identification ◽  
Ixodes Persulcatus ◽  
Wasp Species ◽  
Maldi Tof Ms ◽  
Hard Ticks ◽  
Maldi Tof ◽  
Western Africa ◽  
Tof Ms

Some parasitoids of the genus Ixodiphagus (Hymenoptera, Chalcidoidea: Encyrtidae) are well-known natural enemies of ticks. In this study, we investigate the occurrence of parasitoid wasps in adult hard ticks from Western Africa (Côte d’Ivoire and Senegal) and Far Eastern Europe (Russia) using molecular methods. The morphological identification allowed the classification of 785 collected specimens of six species of ticks: Rhipicephalus (Boophilus) microplus (41%), Ixodes persulcatus (33%), Dermacentor silvarum (11%), Haemaphysalis concinna (7%), Amblyomma variegatum (5%), and Haemaphysalis japonica (3%). The newly developed MALDI-TOF MS protocol identified tick species in spite of their different storage (dried or in 70% ethanol) conditions for a long period. Molecular screening of ticks by a new standard PCR system developed in this study revealed the presence of parasitoid wasp DNA in 3% (28/785) of analyzed ticks. Ixodiphagus hookeri was detected in 86% (24/28) of infested ticks, including 13 I. persulcatus, 9 R (B) microplus, and one H. concinna and D. silvarum. While an unidentified parasitoid wasp species from the subfamily Aphidiinae and Braconidae family was detected in the remaining 14% (4/28) infested ticks. These infested ticks were identified as I. persulcatus. Our findings highlight the need for further studies to clarify the species diversity of parasitoid infesting ticks by combining molecular and morphological features. The novel molecular and MALDI-TOF MS protocols could be effective tools for the surveillance and characterization of these potential bio-control agents of ticks.

Rapid Identification of Commercial Frankincense Products by MALDI-TOF Mass Spectrometry

2021 ◽  
Vol 24 ◽  
Author(s):  
Shang-Chih Lai ◽  
Ren-In You ◽  
Tz-Ting Chen ◽  
Yu Chang ◽  
Chao-Zong Liu ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Throughput ◽  
High Throughput Screening ◽  
Large Scale ◽  
Reference Sample ◽  
Maldi Tof Ms ◽  
Boswellic Acids ◽  
Maldi Tof ◽  
Tof Ms

Background: Frankincense is a resin secreted by the Boswellia tree. It is used in perfumery, aromatherapy, skincare, and traditional Chinese medicine. However, all Boswellia species are under threat owing to habitat loss and overexploitation. As a result, the market is getting flooded with counterfeit frankincense products. Objective: This study aims to establish a high-throughput method to screen and identify the authenticity of commercial frankincense products. We report, for the first time, a matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS)-based method for rapid and high-throughput screening of frankincense samples. Methods: MALDI-TOF MS, HPLC, thin-layer chromatography (TLC), and in vitro anti-inflammatory activity assay were used to examine the frankincense samples. Results: Well-resolved peaks of frankincense triterpenoids in the spectra were observed in the crude extract of commercial samples, including α-boswellic acids (αBAs), β-boswellic acids (βBAs), 11-keto-β-boswellic acids (KBAs), acetyl-11-keto-β-boswellic acids (AKBAs), and their esters. These compounds can be used as indicators for determining the authenticity of frankincense. Conclusion: Unlike LC–MS, which is a time-consuming and expensive method, and TLC, which requires a reference sample, our inexpensive, rapid high-throughput identification method based on MALDI-TOF MS is ideal for large-scale screening of frankincense samples sold in the market.

scholarly journals Rapid and reproducible MALDI-TOF-based method for detection Vancomycin-resistant Enterococcus faecium using classifying algorithms

2021 ◽  
Author(s):  
Ana Candela ◽  
Manuel J Arroyo ◽  
Angela Sanchez-Molleda ◽  
Gema Méndez ◽  
David Rodriguez-Temporal ◽  
...  
Keyword(s):  
Enterococcus Faecium ◽  
Principal Component ◽  
Resistance Mechanisms ◽  
Support Vector ◽  
Accurate Identification ◽  
Maldi Tof Ms ◽  
Maldi Tof ◽  
Vancomycin Resistant ◽  
Lower Variability ◽  
Tof Ms

Vancomycin-resistant Enterococcus faecium has become a health threat over the last 20 years due to its ability to rapidly spread and cause outbreaks in hospital settings. Although MALDI-TOF MS has already demonstrated its usefulness for accurate identification of E. faecium, its implementation for antimicrobial resistance detection is still under evaluation. The reproducibility of MALDI-TOF MS for peak analysis and its performance for correct discrimination of vancomycin susceptible isolates (VSE) from those hosting the VanA and VanB resistance mechanisms was evaluated in this study. For the first goal, intra-spot, inter-spot -technical- and inter-day -biological- reproducibility was assayed. The capability of MALDI-TOF to discriminate VSE isolates from VanA VRE and VanB VRE strains was carried out on protein spectra from 178 E. faecium unique clinical isolates -92 VSE, 31 VanA VRE, 55 VanB VRE-, processed with Clover MS Data Analysis software. Unsupervised (Principal Component Analysis –PCA-) and supervised algorithms (Support Vector Machine -SVM-, Random Forest -RF- and Partial Least Squares-Discriminant Analysis -PLS-DA-) were applied. The reproducibility assay showed lower variability for normalized data (p<0.0001) and for the peaks within the 3000-9000 m/z range. Besides, 80.9%, 79.21% and 77.53% VSE vs VRE (VanA + VanB) discrimination was achieved by applying SVM, RF and PLS-DA, respectively. Correct differentiation of VanA from VanB VRE isolates was obtained by SVM in 86.65% cases. The implementation MALDI-TOF MS and peak analysis could represent a rapid and effective tool for VRE screening. However, further improvements are needed to increase the accuracy of this approach.

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