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      Proteomic characterization and discrimination of Aeromonas species recovered from meat and water samples with a spotlight on the antimicrobial resistance of Aeromonas hydrophila

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          Abstract

          Aeromonas is recognized as a human pathogen following ingestion of contaminated food and water. One major problem in Aeromonas identification is that certain species are phenotypically very similar. The antimicrobial resistance is another significant challenge worldwide. We therefore aimed to use mass spectrometry technology for identification and discrimination of Aeromonas species and to screen the antimicrobial resistance of Aeromonas hydrophila (A. hydrophila). A total of 150 chicken meat and water samples were cultured, and then, the isolates were identified biochemically by the Vitek ® 2 Compact system. Proteomic identification was performed by MALDI‐TOF MS and confirmed by a microchannel fluidics electrophoresis assay. Principal component analysis (PCA) and single‐peak analysis created by MALDI were also used to discriminate the Aeromonas species. The antimicrobial resistance of the A. hydrophila isolates was determined by Vitek ® 2 AST cards. In total, 43 samples were positive for Aeromonas and comprised 22 A. hydrophila, 12 Aeromonas caviae ( A. caviae), and 9 Aeromonas sobria ( A. sobria) isolates. Thirty‐nine out of 43 (90.69%) Aeromonas isolates were identified by the Vitek ® 2 Compact system, whereas 100% of the Aeromonas isolates were correctly identified by MALDI‐TOF MS with a score value ≥2.00. PCA successfully separated A. hydrophila, A. caviae and A. sobria isolates into two groups. Single‐peak analysis revealed four discriminating peaks that separated A. hydrophila from A. caviae and A. sobria isolates. The resistance of A. hydrophila to antibiotics was 95.46% for ampicillin, 50% for cefotaxime, 45.45% for norfloxacin and pefloxacin, 36.36% for ceftazidime and ciprofloxacin, 31.81% for ofloxacin and 27.27% for nalidixic acid and tobramycin. In conclusion, chicken meat and water were tainted with Aeromonas spp., with a high occurrence of A. hydrophila. MALDI‐TOF MS is a powerful technique for characterizing aeromonads at the genus and species levels. Future studies should investigate the resistance of A. hydrophila to various antimicrobial agents.

          Abstract

          The sequencing results indicated that A. hydrophila is the most prevalent Aeromonas spp. isolated from food and water. MALDI‐TOF MS is a powerful technique used for identification of Aeromonas at the genus and species‐level. Principal component analysis (PCA) and single‐peak analysis are successful tools to discriminate the Aeromonas spp. VITEK ® 2 AST Cards can also use as a detective method of antimicrobial resistance.

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          Ongoing revolution in bacteriology: routine identification of bacteria by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

          Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry accurately identifies both selected bacteria and bacteria in select clinical situations. It has not been evaluated for routine use in the clinic. We prospectively analyzed routine MALDI-TOF mass spectrometry identification in parallel with conventional phenotypic identification of bacteria regardless of phylum or source of isolation. Discrepancies were resolved by 16S ribosomal RNA and rpoB gene sequence-based molecular identification. Colonies (4 spots per isolate directly deposited on the MALDI-TOF plate) were analyzed using an Autoflex II Bruker Daltonik mass spectrometer. Peptidic spectra were compared with the Bruker BioTyper database, version 2.0, and the identification score was noted. Delays and costs of identification were measured. Of 1660 bacterial isolates analyzed, 95.4% were correctly identified by MALDI-TOF mass spectrometry; 84.1% were identified at the species level, and 11.3% were identified at the genus level. In most cases, absence of identification (2.8% of isolates) and erroneous identification (1.7% of isolates) were due to improper database entries. Accurate MALDI-TOF mass spectrometry identification was significantly correlated with having 10 reference spectra in the database (P=.01). The mean time required for MALDI-TOF mass spectrometry identification of 1 isolate was 6 minutes for an estimated 22%-32% cost of current methods of identification. MALDI-TOF mass spectrometry is a cost-effective, accurate method for routine identification of bacterial isolates in or =10 reference spectra per bacterial species and a 1.9 identification score (Brucker system). It may replace Gram staining and biochemical identification in the near future.
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            Bacteria antibiotic resistance: New challenges and opportunities for implant-associated orthopedic infections

            There has been a dramatic increase in the emergence of antibiotic resistant bacterial strains, which has made antibiotic choices for infection control increasingly limited and more expensive. In the U.S. alone, antibiotic resistant bacteria cause at least 2 million infections and 23,000 deaths a year resulting in a $55–70 billion per year economic impact. Antibiotics are critical to the success of surgical procedures including orthopaedic prosthetic surgeries, and antibiotic resistance is occurring in nearly all bacteria that infect people, including the most common bacteria that cause orthopaedic infections, such as Staphylococcus aureus ( S. aureus ). Most clinical cases of orthopaedic surgeries have shown that patients infected with antibiotic resistant bacteria, such as methicillin resistant S. aureus (MRSA), are associated with increased morbidity and mortality. This paper reviews the severity of antibiotic resistance at the global scale, the consequences of antibiotic resistance, and the pathways bacteria used to develop antibiotic resistance. It highlights the opportunities and challenges in limiting antibiotic resistance through approaches like the development of novel, non-drug approaches to reduce bacteria functions related to orthopaedic implant-associated infections.
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              MALDI TOF MS profiling of bacteria at the strain level: a review.

              Since the advent of the use of matrix-assisted laser desorption/ionization (MALDI) time-of-flight mass spectrometry (TOF MS) as a tool for microbial characterization, efforts to increase the taxonomic resolution of the approach have been made. The rapidity and efficacy of the approach have suggested applications in counter-bioterrorism, prevention of food contamination, and monitoring the spread of antibiotic-resistant bacteria. Strain-level resolution has been reported with diverse bacteria, using library-based and bioinformatics-enabled approaches. Three types of characterization at the strain level have been reported: strain categorization, strain differentiation, and strain identification. Efforts to enhance the library-based approach have involved sample pre-treatment and data reduction strategies. Bioinformatics approaches have leveraged the ever-increasing amount of publicly available genomic and proteomic data to attain strain-level characterization. Bioinformatics-enabled strategies have facilitated strain characterization via intact biomarker identification, bottom-up, and top-down approaches. Rigorous quantitative and advanced statistical analyses have fostered success at the strain level with both approaches. Library-based approaches can be limited by effects of sample preparation and culture conditions on reproducibility, whereas bioinformatics-enabled approaches are typically limited to bacteria, for which genetic and/or proteomic data are available. Biological molecules other than proteins produced in strain-specific manners, including lipids and lipopeptides, might represent other avenues by which strain-level resolution might be attained. Immunological and lectin-based chemistries have shown promise to enhance sensitivity and specificity. Whereas the limits of the taxonomic resolution of MALDI TOF MS profiling of bacteria appears bacterium-specific, recent data suggest that these limits might not yet have been reached. Copyright © 2013 Wiley Periodicals, Inc.
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                Author and article information

                Contributors
                aymanella2007@yahoo.com
                Journal
                Microbiologyopen
                Microbiologyopen
                10.1002/(ISSN)2045-8827
                MBO3
                MicrobiologyOpen
                John Wiley and Sons Inc. (Hoboken )
                2045-8827
                06 January 2019
                November 2019
                : 8
                : 11 , ANTIMICROBIAL RESISTANCE ( doiID: 10.1002/mbo3.v8.11 )
                : e782
                Affiliations
                [ 1 ] Department of Bacteriology, Mycology and Immunology, Faculty of Veterinary Medicine University of Sadat City Sadat City Egypt
                [ 2 ] Department of Public Health, College of Public Health and Health Informatics Qassim University Buraidah Saudi Arabia
                [ 3 ] Department of Medical laboratories, College of Applied Medical Science Qassim University Buraidah Saudi Arabia
                [ 4 ] Department of Veterinary Medicine, College of Agriculture and Veterinary Medicine Qassim University Buraidah Saudi Arabia
                [ 5 ] Department of Public Health, College of Applied Medical Science King Khalid University Abha Saudi Arabia
                [ 6 ] Department of Fish Diseases and Management, Faculty of Veterinary Medicine Cairo University Cairo Egypt
                [ 7 ] Department of Food Hygiene & Control, Faculty of Veterinary Medicine University of Sadat City Sadat City Egypt
                [ 8 ] Department of Botany and Microbiology, College of Science King Saud University Riyadh Saudi Arabia
                [ 9 ] Department of Microbiology, Faculty of Veterinary Medicine Cairo University Cairo Egypt
                [ 10 ] Department of Medical Technology/Microbiology, College of Applied Medical Sciences Taibah University Madinah Saudi Arabia
                Author notes
                [*] [* ] Correspondence

                Ayman Elbehiry, Department of Bacteriology, Mycology and Immunology, Faculty of Veterinary Medicine, University of Sadat City, Sadat City, Egypt.

                Email: aymanella2007@ 123456yahoo.com

                Author information
                https://orcid.org/0000-0001-7726-9532
                Article
                MBO3782
                10.1002/mbo3.782
                6854848
                30614207
                9a6b46ec-86ad-4635-8215-19bdf1e622c7
                © 2019 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 13 August 2018
                : 17 November 2018
                : 20 November 2018
                Page count
                Figures: 5, Tables: 5, Pages: 14, Words: 9747
                Funding
                Funded by: Qassim University , open-funder-registry 10.13039/501100007414;
                Categories
                Original Article
                Original Articles
                Custom metadata
                2.0
                November 2019
                Converter:WILEY_ML3GV2_TO_JATSPMC version:5.7.2 mode:remove_FC converted:05.12.2019

                Microbiology & Virology
                aeromonas spp.,antimicrobial resistance,differentiation,microchannel electrophoresis,protein fingerprinting

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