Antibacterial Coatings
The Fight Against Bacteria
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The Importance of Bacteria Identification in Clean Rooms
DOSCH 3D: Medical – Bacteria
Dosch 3D offers 1000s of completely textured 3D-models and scenes for all popular 3D-applications. Users can select from… More >>
Giant Microbes E. coli (Escherichia Coli) Ailmentaries Plush Toy
Dosch 3D offers 1000s of completely textured 3D-models and scenes for all popular 3D-applications. Users can select from… More >>A comprehensive environmental monitoring program of clean rooms should include routine monitoring of both viable and non-viable airborne particulates. Although there is no requirement for the microbial identification of all contaminants present in these controlled environments, an environmental control program shall include an appropriate level of bacteria identification obtained from sampling. There are several methods of bacterial identification available.
The first step for correct bacterial identification, especially concerning a clean room isolate, is the Gram staining, since it can provide elucidative clues about the source of the microbial contamination. If microbial identification of isolates reveals Gram-positive cocci, the source of contamination can be derived from humans. If bacteria identification of isolates reveals Gram-positive rods, the source of contamination can be derived from dust or strains resistant to disinfectants. If bacterial identification of isolates reveals Gram-negative rods, the source of contamination can be derived from water or any moistened surface.
Microbial identification in pharmaceutical clean rooms is required for several reasons associated with quality assurance: determination of organisms from the manufacturing environment; bacteria identification from final product testing; demonstrating absence of named organisms from non-sterile products and water; quality control of fermentation stocks in biotechnology; and confirmation of test organisms in validation processes. More and more, the Food and Drug Administration (FDA) is expecting bacterial identification to aid in determining the usual flora for a specific site, to evaluate the effectiveness of cleaning and to troubleshoot the source of contamination that can occur when action levels are exceeded or sterility tests become contaminated.
Fabio L. C. Pacheco invites you to visit Microbiology Online. If you liked this article, you will also enjoy Microbiology Online, a blog written with the objective to bring you the latest news about microbiology and help professionals from the pharmaceutical industry solve their issues.
Laboratory Guide for Identification of Plant Pathogenic Bacteria
Laboratory Guide for Identification of Plant Pathogenic Bacteria
Identification of Milk Bacteria by Means of the Little Plate
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This is an OCR edition without illustrations or index. It may have numerous typos or missing text. However, purchasers can download a free scanned copy of the original rare book from the publisher’s website (GeneralBooksClub.com). You can also preview excerpts of the book there. Purchasers are also entitled to a free trial membership in the General Books Club where they can select from more than a million books without charge. Original Published by: University of Wi… More >>Identification of Milk Bacteria by Means of the Little Plate
How to Perform Bacteria Identification Using Gas Chromatography of Fatty Acid Methyl Ester?
Ein-O Science BioSigns Bacteria
This hands-on interactive model provides magnified ans cross-sectioned detailing of a Virus. Includes Ein-O’s I Know Gui… More >>
Rising Plague: The Global Threat from Deadly Bacteria and Our Dwindling Arsenal to Fight Them
This hands-on interactive model provides magnified ans cross-sectioned detailing of a Virus. Includes Ein-O’s I Know Gui… More >>As stated in my previous article âThe Importance of Bacteria Identification in Clean Roomsâ, we are supposed to identify the flora obtained from environment samples, and it is not limited to clean room environments. In this article, I am going to focus on bacteria identification solely.
There are several methods of bacteria identification available, which can be classified into two large groups: the phenotypic group and the genotypic group. The phenotypic group includes:
Biochemical tests Biotyping Serotyping Phagetyping Antimicrobial susceptibility Multi-Locus Enzyme Electrophoresis (MLEE) Electrophoretic protein typing and immunoblotting Gas Chromatography of Fatty Acid Methyl Ester
The genotypic group includes:
Plasmid typing Restriction Enzyme Analysis (REA) Pulsed-Field Gel Electrophoresis (PFGE) Ribotyping RAPD Rep-PCR PCR-ribotyping
The only commercially available gas chromatography (GC) system dedicated to bacteria identification by fatty acid methyl ester (FAME) analysis is the Sherlock Microbial Identification System (MIS), developed by Microbial ID, Inc. (MIDI). The original database for aerobic bacteria identification was developed by M. Sasser, in 1990.
The principle of the FAME method rests upon the assumption that some microorganisms have typical cellular fatty acid compositions, which can be compared with the mean fatty acid composition of the strains used to create the library. After comparison, the identities of unknown microorganisms are determined.
For many years, analysis of short chain fatty acids (volatile fatty acids, VFAs) has been routinely used in identification of anaerobic bacteria. In numerous scientific papers, the fatty acids between 9 and 20 carbons in length have also been used for bacteria identification, especially nonfermentative Gram negative organisms. With the advent of fused silica capillary columns (which allows recovery of hydroxy acids and resolution of many isomers), it has become practical and easier to use GC of whole cell fatty acid methyl esters to identify isolated and pure microbial cultures, bacterial of medical importance, and in taxonomic studies.
The FAME method uses a specific sample preparation procedure and a sophisticated chromatographic system to yield qualitatively and quantitatively reproducible fatty acid composition profiles. This system was developed for microbiologists and it does not require extensive knowledge of gas chromatography.
      Sample Preparation Procedure
Bacteria selected for identification by FAME analysis are subcultured twice on Trypticase Soy Broth solidified with 1.5% agar and then incubated aerobically at 28 ºC for 24 h. Growth is examined for the presence of pure culture and submitted to the fatty acid extraction, in simple, five, basic steps.
Harvesting Saponification Methylation Extraction Wash
All glassware should be new and only Teflon and glass should come in contact with reagents.
      Chromatographic System
Recognition of fatty acid profiles is performed using the MIS system along with a standard library. The MIS consists of a gas chromatograph equipped with a fused silica capillary column, a flame ionization detector, an integrator and an automatic sampler coupled to a computer system. The Sherlock computer software automatically sets the operating parameters of the gas chromatograph each time a sample is processed. Fatty acids are separated because of different retention times, using synthetic air, hydrogen as the carrier and nitrogen as the makeup gas. Coupled to Sherlock is the ChemStation software used for operating sampling, analysis, and integration of the chromatographic samples. The fatty acid percentages are automatically calculated and after comparison with the MIDI Standard library, the bacterial identifications are expressed on the basis of genus, species and sub-species level.
Fabio L. C. Pacheco invites you to visit Microbiology Online. If you liked this article, you will also enjoy Microbiology Online. It’s all about great tutorials in industrial microbiology, focusing on pharmaceutical microbiology labs.
Toxigenic Fusarium Species: Identification and Mycotoxicology
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The taxonomy of fungi in the genus Fusarium is complex and confusing because of the different taxonomic systems in use in various parts of the world. Taxonomy of the genus is further complicated by its extreme variability in culture and by the fact that Fusarium species mutate and degenerate rapidly in culture. This situation has led to great confusion in the extensive literature on Fusarium mycotoxicoses because the same fungus is known under a variety of different… More >>Toxigenic Fusarium Species: Identification and Mycotoxicology
Method for the Isolation and Identification of Eccherichia Coli 0157 1996: H7 from Waters
Method for the Isolation and Identification of Eccherichia Coli 0157 1996: H7 from Waters
Kauffmann-White Scheme – 2007: Salmonella Identification; Serotypes and Antigen Formulae
Kauffmann-White Scheme – 2007: Salmonella Identification; Serotypes and Antigen Formulae
Fusarium: A Pictorial Guide to the Identification of Fusarium Species According to the Taxonomic System of Snyder and Hansen
Fusarium: A Pictorial Guide to the Identification of Fusarium Species According to the Taxonomic System of Snyder and Hansen
Pseudomonas syringae Pathovars and Related Pathogens – Identification, Epidemiology and Genomics
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This book reports on recent advances on: (1) new methods and approaches for specific and sensitive detection and identification of Pseudomonas syringae and Ralstonia solanacearum; (2) ecology and epidemiology bases of Pseudomonas syringae that enable the development of management strategies; (3) pathogenesis and determinant of pathogenicity, and in particular, mechanisms involved in virulence and virulence gene expression; (4) evolution and diversity of the pseudomo… More >>Pseudomonas syringae Pathovars and Related Pathogens – Identification, Epidemiology and Genomics
