Antimicrobial resistance of microorganisms from milk and butter and their identification using MALDI-TOF MS Biotyper

Bovine mastitis is a prevalent disease that causes serious economic problems globally in the dairy industry. Staphylococcus aureus is an important pathogen of bovine mastitis. This study was conducted to characterize S. aureus isolates from clinical bovine mastitis cases in large-scale dairy herds in China. S. aureus was isolated from 624 clinical mastitis cases and confirmed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). In total, 62 S. aureus isolates were obtained. Cluster analysis, genetic diversity, quantification of biofilm formation, antimicrobial resistance, and detection of virulence genes were performed on these isolates of S. aureus. Eight isolates harbored the mecA gene and were sensitive to oxacillin. MALDI-TOF MS cluster analysis revealed that the 62 isolates were divided into three major clusters (I, II, III) and eight main groups (A–H) at the distance level of 700. The agr II was the most prevalent (56.5%). The 62 S. aureus isolates were assigned to seven spa types. The most common spa type was t529(58.1%), followed by t2196 (14.5%), t518 (14.5%), t571(6.5%), t034 (3.2%), t2734 (1.6%), and t730 (1.6%). Five STs were identified from seven representative isolates as follows: ST630/CC8, ST97/CC97, ST50, ST398, and ST705. All isolates had the ability to form biofilm. Antimicrobial resistance was most frequently observed to ciprofloxacin (29%), followed by penicillin (24.2%), and streptomycin (9.6%). All isolates harbored the fnbA, clfB (100%), icaA, and icaD genes. This study provides the basis for the development of bovine mastitis prevention program on large-scale dairy farms.

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MALDI-TOF Mass Spectrometry and Specific Biomarkers: Potential New Key for Swift Identification of Antimicrobial Resistance in Foodborne Pathogens

Microorganisms ◽

10.3390/microorganisms7120593 ◽

2019 ◽

Vol 7 (12) ◽

pp. 593 ◽

Cited By ~ 2

Author(s):

Maureen Feucherolles ◽

Henry-Michel Cauchie ◽

Christian Penny

Keyword(s):

Mass Spectrometry ◽

Antimicrobial Resistance ◽

Foodborne Pathogens ◽

Reference Method ◽

Multi Drug Resistance ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Flight Mass Spectrometry ◽

The Future ◽

Tof Ms

Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) is today the reference method for direct identification of microorganisms in diagnostic laboratories, as it is notably time- and cost-efficient. In the context of increasing cases of enteric diseases with emerging multi-drug resistance patterns, there is an urgent need to adopt an efficient workflow to characterize antimicrobial resistance (AMR). Current approaches, such as antibiograms, are time-consuming and directly impact the “patient-physician” workflow. Through this mini-review, we summarize how the detection of specific patterns by MALDI-TOF MS, as well as bioinformatics, become more and more essential in research, and how these approaches will help diagnostics in the future. Along the same lines, the idea to export more precise biomarker identification steps by MALDI-TOF(/TOF) MS data towards AMR identification pipelines is discussed. The study also critically points out that there is currently still a lack of research data and knowledge on different foodborne pathogens as well as several antibiotics families such as macrolides and quinolones, and many questions are still remaining. Finally, the innovative combination of whole-genome sequencing and MALDI-TOF MS could be soon the future for diagnosis of antimicrobial resistance in foodborne pathogens.

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Rapid microbial identification and colistin resistance detection via MALDI-TOF MS using a novel on-target extraction of membrane lipids

Scientific Reports ◽

10.1038/s41598-020-78401-3 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Matthew Sorensen ◽

Courtney E. Chandler ◽

Francesca M. Gardner ◽

Salma Ramadan ◽

Prasanna D. Khot ◽

...

Keyword(s):

Antimicrobial Resistance ◽

Lipid Extraction ◽

Fungal Species ◽

Steel Matrix ◽

Maldi Tof Ms ◽

Microbial Identification ◽

Maldi Tof ◽

Lipid Biomarker ◽

Biomarker Analysis ◽

Tof Ms

AbstractRapid infection diagnosis is critical to improving patient treatment and outcome. Recent studies have shown microbial lipids to be sensitive and selective biomarkers for identifying bacterial and fungal species and antimicrobial resistance. Practical procedures for microbial lipid biomarker analysis will therefore improve patient outcomes and antimicrobial stewardship. However, current lipid extraction methods require significant hands-on time and are thus not suited for direct adoption as a clinical assay for microbial identification. Here, we have developed a method for lipid extraction directly on the surface of stainless-steel matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) plates, termed fast lipid analysis technique or FLAT, which facilitates the identification of bacterial and fungal species using a sub-60-minute workflow. Additionally, our method detects lipid A modifications in Gram-negative bacteria that are associated with antimicrobial resistance, including to colistin.

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Detection of antimicrobial resistance-associated proteins by titanium dioxide-facilitated intact bacteria mass spectrometry

Chemical Science ◽

10.1039/c7sc04089j ◽

2018 ◽

Vol 9 (8) ◽

pp. 2212-2221 ◽

Cited By ~ 20

Author(s):

Yingdi Zhu ◽

Natalia Gasilova ◽

Milica Jović ◽

Liang Qiao ◽

Baohong Liu ◽

...

Keyword(s):

Mass Spectrometry ◽

Titanium Dioxide ◽

Antimicrobial Resistance ◽

Protein Detection ◽

Maldi Tof Ms ◽

Bacteria Identification ◽

Maldi Tof ◽

Associated Proteins ◽

Tof Ms

TiO2-facilitated MALDI–TOF-MS was proposed to improve intact bacteria fingerprinting, allowing rapid and convenient antimicrobial resistance-associated protein detection during bacteria identification.

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Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry for the Rapid Detection of Antimicrobial Resistance Mechanisms and Beyond

Clinical Microbiology Reviews ◽

10.1128/cmr.00037-18 ◽

2018 ◽

Vol 32 (1) ◽

Cited By ~ 24

Author(s):

Marina Oviaño ◽

Germán Bou

Keyword(s):

Antimicrobial Resistance ◽

Laser Desorption ◽

Laser Desorption Ionization ◽

Ionization Time ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Flight Mass Spectrometry ◽

Tof Ms ◽

Matrix Assisted Laser ◽

Matrix Assisted Laser Desorption

SUMMARYMatrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) has been successfully applied in recent years for first-line identification of pathogens in clinical microbiology because it is simple to use, rapid, and accurate and has economic benefits in hospital management. The range of clinical applications of MALDI-TOF MS for bacterial isolates is increasing constantly, from species identification to the two most promising applications in the near future: detection of antimicrobial resistance and strain typing for epidemiological studies. The aim of this review is to outline the contribution of previous MALDI-TOF MS studies in relation to detection of antimicrobial resistance and to discuss potential future challenges in this field. Three main approaches are ready (or almost ready) for clinical use, including the detection of antibiotic modifications due to the enzymatic activity of bacteria, the detection of antimicrobial resistance by analysis of the peak patterns of bacteria or mass peak profiles, and the detection of resistance by semiquantification of bacterial growth in the presence of a given antibiotic. This review provides an expert guide for MALDI-TOF MS users to new approaches in the field of antimicrobial resistance detection, especially possible applications as a routine diagnostic tool in microbiology laboratories.

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Differences in sheep and goats milk microbiological profile between conventional and organic farming systems in Greece

Journal of Dairy Research ◽

10.1017/s0022029917000103 ◽

2017 ◽

Vol 84 (2) ◽

pp. 206-213 ◽

Cited By ~ 3

Author(s):

Eleni Malissiova ◽

Theofilos Papadopoulos ◽

Aikaterini Kyriazi ◽

Maria Mparda ◽

Christina Sakorafa ◽

...

Keyword(s):

Antimicrobial Resistance ◽

Organic Farming ◽

Farming Practices ◽

Organic Farms ◽

Maldi Tof Ms ◽

E Coli ◽

Microbiological Profile ◽

Maldi Tof ◽

Conventional Farms ◽

Tof Ms

The aim of this study was to examine differences in the microbiological profile and antimicrobial resistance of bacteria isolated from milk from organic and conventional sheep and goat farms. Twenty-five organic and 25 conventional sheep and goat farms in the region of Thessaly, Greece participated in this study. A standardised detailed questionnaire was used to describe farming practices. A total of 50 samples were collected and analysed for total viable count (TVC), total coliform count (TCC) and somatic cell count (SCC), whileStaphylococcus aureusandEscherichia coliwere isolated using standard methods. Isolates were identified at species level by Api-test and Matrix-Assisted Laser Desorption/Ionisation-Time of Flight Mass Spectrometry (MALDI-TOF MS). Susceptibility to a panel of 20 forE. coliand 16 forS. aureusantimicrobials was determined by the agar dilution method. Pulsed Field Gel Electrophoresis (PFGE) was performed forS. aureusandE. coliisolates to determine predominant clones. Lower counts of TVC, TCC and SCC were identified in milk from the organic farms, possibly due to differences in the hygienic farming practices found on those farms. API-tests and MALDI-TOF MS showed no significant differences in theS. aureusandE. coliisolates. Overall, antimicrobial resistance rates were low, while a statistically higher percentage was estimated among strains originating from conventional farms in comparison with organic farms, possibly due to the restriction of antibiotic use in organic farming. PFGE revealed diversity amongS. aureusandE. colipopulations in both organic and conventional farms indicating circulation of 2–3 main clones changing slightly during their evolution. Consequently, there is evidence that milk from the organic farms presents a better microbiological profile when compared with milk from conventional farms.

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MALDI-TOF Mass Spectrometry Technology as a Tool for the Rapid Diagnosis of Antimicrobial Resistance in Bacteria

Antibiotics ◽

10.3390/antibiotics10080982 ◽

2021 ◽

Vol 10 (8) ◽

pp. 982

Author(s):

Eun-Jeong Yoon ◽

Seok Hoon Jeong

Keyword(s):

Mass Spectrometry ◽

Antimicrobial Resistance ◽

Low Cost ◽

Rapid Identification ◽

Diagnostic Process ◽

Antimicrobial Drugs ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Mass Spectrometry Technology ◽

Tof Ms

Species identification by using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a routine diagnostic process for infectious diseases in current clinical settings. The rapid, low-cost, and simple to conduct methodology is expanding its application in clinical microbiology laboratories to diagnose the antimicrobial resistance (AMR) in microorganisms. Primarily, antimicrobial susceptibility testing is able to be carried out either by comparing the area under curve of MALDI spectra of bacteria grown in media with antimicrobial drugs or by identifying the shift peaks of bacteria grown in media including 13C isotope with antimicrobial drugs. Secondly, the antimicrobial resistance is able to be determined through identifying (i) the antimicrobial-resistant clonal groups based on the fingerprints of the clone, (ii) the shift peak of the modified antimicrobial drug, which is inactivated by the resistance determinant, (iii) the shift peak of the modified antimicrobial target, (iv) the peak specific for the antimicrobial determinant, and (v) the biomarkers that are coproduced proteins with AMR determinants. This review aims to present the current usage of the MALDI-TOF MS technique for diagnosing antimicrobial resistance in bacteria, varied approaches for AMR diagnostics using the methodology, and the future applications of the methods for the accurate and rapid identification of AMR in infection-causing bacterial pathogens.

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