Lect3_Slides - Cultivating and Identifying Microbes...

Info iconThis preview shows page 1. Sign up to view the full content.

View Full Document Right Arrow Icon
This is the end of the preview. Sign up to access the rest of the document.

Unformatted text preview: Cultivating and Identifying Microbes Outline: HOW? 1. Choice of environmental sample 2. Analyzing the sample microscopically - Gram Stain 3. Enrichment Growth medium Liquid vs Solid 4. Isolating a pure culture – streak purification 5. Identifying the bacterium – DNA fingerprinting 6. Estimate recovery of microbial cells from a sample WHY? Gram Stain Neisseria meningiditis •  Figure legend: 
 Gram stain of liquor of a patient with meningococcal meningitis. Nuclei and cell membrane outlines of many polymorphonuclear cells are seen as well as intracellular gram-negative diplococci (Neisseria meningitidis). (http://www.genomik.uni-wuerzburg.de/pathogenomik-plus_2006__2009/research_groups/cluster_2/) Liquid Versus Solid Medium DNA Fingerprinting 1. Isolate chromosomal DNA 2. Digest with a restriction enzyme 3. Separate DNA fragments with PFGE cartoon animation PFGE N. meningitidis Isolates Lanes 1, 6 & 10 = MW markers 2 = Nm case 1 3 = Nm case 2 4 = Nm case 3 5, 7-9 = control Nm For your amusement Patterns on pulsed-field gel electrophoresis of Neisseria meningitidis restricted by SpeI. Lanes 1, 6, 10: lambda ladder reference. Lanes 2, 3, 4: N meningitidis isolates from Cases 1, 2, and 3, respectively. Lanes 5, 7, 8, 9: N meningitidis control isolates from 1999.) The three case patients had all been previously healthy and had not attended college. A search for a common exposure among the three revealed that all had attended a party on May 14, 1999, which was also attended by at least 80 other persons, most of whom were 18 to 24 years old. At the party, a large number of persons reportedly smoked tobacco, smoked marijuana, and/or drank alcohol. The only exposure common to all three patients was the party. Although the patients in case 1 and case 2 were household roommates, the patient in case 3 had no contact with the other two, either before or after the party. Antimicrobial chemoprophylaxis was recommended to a total of 83 party, work, and household contacts of the patients. www.medscape.com/ viewarticle/423567_3 Viable Cell Count Direct Microscopic Cell Count ...
View Full Document

This note was uploaded on 11/23/2009 for the course MIMG 101 taught by Professor Bradley during the Winter '08 term at UCLA.

Ask a homework question - tutors are online