Occurrence of Listeria monocytogenes in Artisanal Cheeses from Poland and Its Identification by MALDI-TOF MS.
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SUBMITTER: Pyz-Lukasik R
PROVIDER: S-EPMC8224308 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:Artisanal cheeses can be sources of Listeria monocytogenes and cause disease in humans. This bacterial pathogen is a species of diverse genotypic and phenotypic characteristics. The aim of the study was to characterize 32 isolates of L. monocytogenes isolated in 2014-2018 from artisanal cheeses. The isolates were characterized using whole genome sequencing and bioinformatics analysis. The artisanal cheese isolates resolved to four molecular groups: 46.9% of them to IIa (1/2a-3a), 31.2% to IVb (4ab-4b-4d-4e), 12.5% to IIc (1/2c-3c), and 9.4% to IIb (1/2b-3b-7). Two evolutionary lineages emerged: lineage II having 59.4% of the isolates and lineage I having 40.6%. The sequence types (ST) totaled 18: ST6 (15.6% of the isolates), ST2, ST20, ST26, and ST199 (each 9.4%), ST7 and ST9 (each 6.3%), and ST1, ST3, ST8, ST16, ST87, ST91, ST121, ST122, ST195, ST217, and ST580 (each 3.1%). There were 15 detected clonal complexes (CC): CC6 (15.6% of isolates), CC9 (12.5%), CC2, CC20, CC26, and CC199 (each 9.4%), CC7 and CC8 (each 6.3%), and CC1, CC3, CC14, CC87, CC121, CC195, and CC217 (each 3.1%). The isolates were varied in their virulence genes and the differences concerned: inl, actA, LIPI-3, ami, gtcA, aut, vip, and lntA.
| S-EPMC9497517 | biostudies-literature
Project description:Truffles are edible mushrooms with similar morphological characteristics, that make it difficult to distinguish between highly prized truffles (such as the Périgord black T. melanosporum) and inexpensive truffles (such as the Asian Black T. indicum). These biological and economic features have led to several misidentifications and/or fraudulent profit in the truffle markets. In this paper, we investigate Matrix-assisted Laser Desorption/Ionization Time-Of-Flight Mass Spectrometry (MALDI-TOF MS) biotyping to identify 34 commercial fresh truffles from Europe and Asia. The MALDI-TOF MS clustering rapidly distinguished seven Tuber species identified by ITS phylogenetic analysis. The tasty T. melanosporum was clearly differentiated from the Chinese and less expensive truffles. These cheaper mushrooms were marketed as T. indicum but corresponded to a mix of three species. In total, the method confirmed misidentifications in 26% of commercial specimens. Several unknown blind-coded truffles were rapidly identified, with scores >= 2, using the Bruker Biotyper algorithm against MS databases. This study demonstrates that MALDI-TOF MS is a reliable, rapid and cheaper new tool compared with molecular methods for the identification of truffle species and could be used to control frauds in the truffle markets. It could also be useful for the certification of truffle-inoculated seedlings and/or diversity in forest ecosystems.
| S-EPMC6881316 | biostudies-literature
Project description:The genetic lineages of Listeria monocytogenes and other species of the genus Listeria are correlated with pathogenesis in humans. Although matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has become a prevailing tool for rapid and reliable microbial identification, the precise discrimination of Listeria species and lineages remains a crucial issue in clinical settings and for food safety. In this study, we constructed an accurate and reliable MS database to discriminate the lineages of L. monocytogenes and the species of Listeria (L. monocytogenes, L. innocua, L. welshimeri, L. seeligeri, L. ivanovii, L. grayi, and L. rocourtiae) based on the S10-spc-alpha operon gene encoded ribosomal protein mass spectrum (S10-GERMS) proteotyping method, which relies on both genetic information (genomics) and observed MS peaks in MALDI-TOF MS (proteomics). The specific set of eight biomarkers (ribosomal proteins L24, L6, L18, L15, S11, S9, L31 type B, and S16) yielded characteristic MS patterns for the lineages of L. monocytogenes and the different species of Listeria, and led to the construction of a MS database that was successful in discriminating between these organisms in MALDI-TOF MS fingerprinting analysis followed by advanced proteotyping software Strain Solution analysis. We also confirmed the constructed database on the proteotyping software Strain Solution by using 23 Listeria strains collected from natural sources.
| S-EPMC4956195 | biostudies-literature
Project description:BackgroundIn the last decade, an innovative approach has emerged for arthropod identification based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Increasing interest in applying the original technique for arthropod identification has led to the development of a variety of procedures for sample preparation and selection of body parts, among others. However, the absence of a consensual strategy hampers direct inter-study comparisons. Moreover, these different procedures are confusing to new users. Establishing optimized procedures and standardized protocols for mosquito identification by MALDI-TOF MS is therefore a necessity, and would notably enable the sharing of reference MS databases. Here, we assess the optimal conditions for mosquito identification using MALDI-TOF MS profiling.MethodsThree homogenization methods, two of which were manual and one automatic, were used on three distinct body parts (legs, thorax, head) of two mosquito laboratory strains, Anopheles coluzzii and Aedes aegypti, and the results evaluated. The reproducibility of MS profiles, identification rate with relevant scores and the suitability of procedures for high-throughput analyses were the main criteria for establishing optimized guidelines. Additionally, the consequences of blood-feeding and geographical origin were evaluated using both laboratory strains and field-collected mosquitoes.ResultsRelevant score values for mosquito identification were obtained for all the three body parts assayed using MALDI-TOF MS profiling; however, the thorax and legs were the most suitable specimens, independently of homogenization method or species. Although the manual homogenization methods were associated with a high rate of identification on the three body parts, this homogenization mode is not adaptable to the processing of a large number of samples. Therefore, the automatic homogenization procedure was selected as the reference homogenization method. Blood-feeding status did not hamper the identification of mosquito species, despite the presence of MS peaks from original blood in the MS profiles of the three body parts tested from both species. Finally, a significant improvement in identification scores was obtained for field-collected specimens when MS spectra of species from the same geographical area were added to the database.ConclusionThe results of the current study establish guidelines for the selection of mosquito anatomic parts and modality of sample preparation (e.g. homogenization) for future specimen identification by MALDI-TOF MS profiling. These standardized operational protocols could be used as references for creating an international MS database.
| S-EPMC9248115 | biostudies-literature
Project description:Identification of different Trichuris spp. using MALDI-TOF MS
| PRJEB50468 | ENA
Project description:BackgroundAnopheles species identification is essential for an effective malaria vector control programme. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) has been developed to identify adult Anopheles species, using the legs or the cephalothorax. The protein repertoire from arthropods can vary according to compartment, but there is no general consensus regarding the anatomic part to be used.MethodsTo determine the body part of the Anopheles mosquitoes best suited for the identification of field specimens, a mass spectral library was generated with head, thorax with wings and legs of Anopheles gambiae, Anopheles arabiensis and Anopheles funestus obtained from reference centres. The MSL was evaluated using two independent panels of 52 and 40 An. gambiae field-collected in Mali and Guinea, respectively. Geographic variability was also tested using the panel from Mali and several databases containing added specimens from Mali and Senegal.ResultsUsing the head and a database without specimens from the same field collection, the proportion of interpretable and correct identifications was significantly higher than using the other body parts at a threshold value of 1.7 (p?<?0.0001). The thorax of engorged specimens was negatively impacted by the blood meal after frozen storage. The addition of specimens from Mali into the database significantly improved the results of Mali panel (p?<?0.0001), which became comparable between head and legs. With higher identification scores, the using of the head will allow to decrease the number of technical replicates of protein extract per specimen, which represents a significant improvement for routine use of MALDI-TOF MS.ConclusionsThe using of the head of Anopheles may improve the performance of MALDI-TOF MS. Region-specific mass spectrum databases will have to be produced. Further research is needed to improve the standardization in order to share online spectral databases.
| S-EPMC7796594 | biostudies-literature
Project description:BACKGROUND:Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been rapidly developed and widely used as an analytical technique in clinical laboratories with high accuracy in microorganism identification. OBJECTIVE:To validate the efficacy of MALDI-TOF MS in identification of clinical pathogenic anaerobes. METHODS:Twenty-eight studies covering 6685 strains of anaerobic bacteria were included in this meta-analysis. Fixed-effects models based on the P-value and the I-squared were used for meta-analysis to consider the possibility of heterogeneity between studies. Statistical analyses were performed by using STATA 12.0. RESULTS:The identification accuracy of MALDI-TOF MS was 84% for species (I2?=?98.0%, P?<?0.1), and 92% for genus (I2?=?96.6%, P?<?0.1). Thereinto, the identification accuracy of Bacteroides was the highest at 96% with a 95% CI of 95-97%, followed by Lactobacillus spp., Parabacteroides spp., Clostridium spp., Propionibacterium spp., Prevotella spp., Veillonella spp. and Peptostreptococcus spp., and their correct identification rates were all above 90%, while the accuracy of rare anaerobic bacteria was relatively low. Meanwhile, the overall capabilities of two MALDI-TOF MS systems were different. The identification accuracy rate was 90% for VITEK MS vs. 86% for MALDI biotyper system. CONCLUSIONS:Our research showed that MALDI-TOF-MS was satisfactory in genus identification of clinical pathogenic anaerobic bacteria. However, this method still suffers from different drawbacks in precise identification of rare anaerobe and species levels of common anaerobic bacteria.
| S-EPMC6836477 | biostudies-literature
Project description:Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS) has proved effective for the identification of many arthropods. A total of 432 termite specimens were collected in Mali, Cote d'Ivoire, Togo, Senegal, Switzerland and France. Morphologically, 22 species were identified, including Ancistrotermes cavithorax, Amitermes evuncifer, Cryptotermes brevis, Cubitermes orthognathus, Kalotermes flavicollis, Macrotermes bellicosus, Macrotermes herus, Macrotermes ivorensis, Macrotermes subhyalinus, Microcerotermes parvus, Microtermes sp., Odontotermes latericius, Procubitermes sjostedti, Promirotermes holmgreni, Reticulitermes grassei, Reticulitermes lucifugus, Reticulitermes santonensis, Trinervitermes geminatus, Trinervitermes occidentalis, Trinervitermes togoensis, Trinervitermes sp., Trinervitermes trinervoides and Trinervitermes trinervius. Analysis of MALDI-TOF MS spectra profiles from termites revealed that all were of high quality, with intra-species reproducibility and inter-species specificity. Blind testing of the spectra of 389 termites against our updated database with the spectra of 43 specimens of different termite species revealed that all were correctly identified with log score values (LSVs) ranging from 1.65 to 2.851, mean 2.290 ± 0.225, median 2.299, and 98.4% (383) had LSVs > 1.8. This study is the first on the use of MALDI-TOF for termite identification and shows its importance as a tool for arthropod taxonomy and reinforces the idea that MALDI-TOF MS is a promising tool in the field of entomology.
| S-EPMC8760289 | biostudies-literature
Project description:Microorganism identification by MALDI TOF mass-spectrometry is based on the comparison of the mass spectrum of the studied organism with those of reference strains. It is a rapid and reliable method. However, commercial databases and programs are mostly designed for identification of clinically important strains and can be used only for particular mass spectrometer models. The need for open platforms and reference databases is obvious. In this study we describe a geometric approach for microorganism identification by mass spectra and demonstrate its capabilities by analyzing 24 strains belonging to the Bacillus pumilus group. This method is based on representing mass spectra as points on a multidimensional space, which allows us to use geometric distances to compare the spectra. Delimitation of microorganisms performed by geometric approach correlates well with the results of molecular phylogenetic analysis and clustering using Biotyper 3.1. All three methods used allowed us to reliably divide the strains into two groups corresponding to closely related species, Bacillus pumilus and Bacillus altitudinis. The method developed by us will be implemented in a Web interface designed for using open reference databases for microorganism identification. The data is available at http://www.bionet.nsc.ru/mbl/database/database.html.
| S-EPMC4655313 | biostudies-literature
Project description:Prenatal diagnosis aims either to provide the reassurance to the couples at risk of having an affected child by timely appropriate therapy or to give the parents a chance to decide the fate of the unborn babies with health problems. Invasive prenatal diagnosis (IPD) is accurate, however, carrying a risk of miscarriage. Non-invasive prenatal diagnosis (NIPD) has been developed based on the existing of fetal genetic materials in maternal circulation; however, a minority fetal DNA in majority maternal background DNA hinders the detections of fetal traits. Different protocols and assays, such as homogenous MassEXTEND (hME), single allele base extension reaction (SABER), precise measuring copy number variation of each allele, and quantitative methylation and expression analysis using the high-throughput sensitive matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS), allow NIPD for single gene disorders, fetal blood group genotyping and fetal aneuploidies as well as the development of fetal gender-independent biomarkers in maternal circulation for management of pathological pregnancies. In this review, we summarise the use of MALDI-TOF MS in prenatal genomics.
| S-EPMC2941831 | biostudies-literature