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Microbiology discussion… Virus Vs Bacteria ID / Inspection Techniques Bacteria A bacterium is a microscopic single cell organism Found anywhere: – Air – Soil – Water – Plants and Animals. Function: Bacteria There are good & bad bacteria. They have useful functions: – making vitamins – break down garbage The human mouth is home to more than 500 species of Bacteria. 4 Culturing Bacteria Culturing Bacteria The Six “I’s” – Inoculation: Introduce bacteria to culture Incubation: Provide optimal growing conditions. Isolation: Separating a “type” of bacteria from the culture Inspection: Stain & study bacteria for shape, size & color *Info Gathering: at this point we will skip this step Identification: ID the bacterial Gram + or Gram –; Coccus, bacillus, or sprilla Gram + or Gram - / Shape The Gram Stain / revisited Copyright © McGraw-Hill Education. Permission required for reproduction or display. Microscopic Appearance of Cell Step 1 Crystal Violet (primary dye) 2 Gram’ siodine (mordant) Gram (+) Gram (–) Chemical Reaction in Cell (very magnified view) Gram (+) Gram (–) Both cell walls stain with the dye. Dye crystals trapped in cell No effect of iodine 3 Alcohol (decolorizer) 4 Safranin (red dye counterstain) Crystals remain in cell. Outer wall is weakened; cell loses dye. Red dye has no effect. Red dye stains the colorless cell. 8 Virus Viruses are the simplest of microbes. They can be as much as 10,000 times smaller than Bacteria Viruses are made up of DNA or RNA Viral Shapes Function Virus When viruses come into contact with the host cell they hijack the cell & release their DNA into the cell. It can take over immediately or lie dormant for years The host cell does all the work, the viruses just gives the orders Culturing Viruses Culturing Viruses – Animal viruses may be cultured using a variety of means Whole animals Eggs Tissue culture – Tissue Culture used today for most viruses & vaccines – This involves growing animal cells in flasks using infecting these cells with virus FYI - Vaccine Selection The viruses used in making flu vaccine are chosen each year on information from the year before Info is gathered by 94 countries and analyzed by: – World Health Organization (WHO) – The Centers for Disease Control (CDC): Atlanta, Georgia London, UK Melbourne, Australia Tokyo, Japan Experts forecast which viruses are likely to circulate the following season 2011-2012 Flu Vaccines The flu vaccine protects against the three main flu strains that research indicates will cause the most illness during the flu season. They are: – A/California/7/2009 (H1N1)-like virus – A/Perth/16/2009 (H3N2)-like virus – B/Brisbane/60/2008-like virus 2012-2013 / 2013-2014 A/California/7/2009 (H1N1)pdm09-like virus A/Victoria/361/2011 (H3N2)-like virus B/Wisconsin/1/2010-like virus (from the B/Yamagata lineage of viruses = sub-type of Type B flu – the most common type) an A/California/7/2009 (H1N1)pdm09-like virus; an A(H3N2) virus antigenically like the cellpropagated prototype virus A/Victoria/361/2011; a B/Massachusetts/2/2012-like (B/Yamagata lineage) virus. 2015 - 2016 an A/California/7/2009 (H1N1)pdm09-like virus an A/Switzerland/9715293/2013 (H3N2)-like virus a B/Phuket/3073/2013-like virus. (This is a B/Yamagata lineage virus) – Some of the 2015-2016 flu vaccine is quadrivalent vaccine and also protects against an additional B virus (B/Brisbane/60/2008-like virus). This is a B/Victoria lineage virus. Parts of the Microscope 22 Magnification power Eye piece: 10X – Low: – Medium: – High: 4X = 10X = 40X = 10 x 4 = 40 10 x 10 = 100 10 x 40 = 400 Resolution The capacity to distinguish or separate two objects 24 The Purpose of Oil 25 2 Types of Electron Microscopes Transmission electron microscopes – (TEM) transparent. 26 2 Types of Electron Microscopes Scanning electron microscopes – (SEM) –3D view. SEM bombards surface of metalcoated specimen with electrons while scanning back and forth 27 FEW EXAMPLES OF MEDIA The next few slides have some culture media examples You have already seen some of these; but now that you have worked with it, it may mean more to see it again. Inspection If a single species is growing in the container, you have a pure culture but if there are multiple species than you have a mixed culture. Check for contaminants (unknown or unwanted microbes) in the culture. 29 Physical States of Media Liquid – broth; does not solidify Semisolid – contains solidifying agent Solid – firm surface for colony formation 30 Gram Staining http://www.youtube.com/watch?v=O Q6C-gj_UHM