Price: $21.95
Thursday, October 14, 2010
Microbiology (Flash Cards)
Lambda Phage as a Model Organism [VHS]
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Dr. Mary's Monkey: How the Unsolved Murder of a Doctor, a Secret Laboratory in New Orleans and Cancer-Causing Monkey Viruses are Linked to Lee Harvey Oswald, ... Assassination and Emerging Global Epidemics
Price: $19.95
Wednesday, October 13, 2010
Key to Successful "in Situ" Pcr [VHS]
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Immunology in Focus [VHS]
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Immunology in Focus [VHS]
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Tuesday, October 12, 2010
2000 and Beyond: Confronting the Microbe Menace (Four Volumes)
Tape 1: Microbe Hunters: Tracking Infectious Agents
Tape 2: The Microbes Strike Back
Tape 3: Outwitting Bacteria's Wily Ways
Tape 4: Emerging Infections: How Epidemics Arise
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PAL Video:HIV & AIDS Set [VHS]
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Monday, October 11, 2010
Microcards: Review Cards for Medical Students
These flashcards will help medical students organize and recall medical microbiology information for course exams and USMLE Step 1. The cards are grouped into sections on gram-positive, gram-negative, and other bacteria; RNA and DNA viruses; fungi; protozoa; and helminths. Each section begins with charts summarizing key information on the group of microorganisms, followed by cards for each individual microbe, which include clinical presentation, pathobiology, diagnosis/treatment, quick facts, and a case study. Schematic illustrations show the morphology and pathogenesis of different microorganisms.
A companion Website provides 70 USMLE-style questions and answers.
Price: $36.95
Clinical Microbiology Made Ridiculously Simple (Medmaster)
Price: $32.95
Friday, October 1, 2010
Gram Staining
The procedure adopted in our institution is as follows:
The smear is covered with few drops of crystal violet solution and allowed to act for one minute. The slide is then washed with gentle stream of running tap water. The smear is then covered with few drops of Gram’s Iodine and allowed to act for a minute and then washed in tap water. The smear is then decolourised by alcohol by until no more violet colour comes off the slide. This process is completed within 30 seconds to prevent overdecolourisation. The slide is washed in water and counterstained using dilute carbol fuchsin for 30 seconds. The slide is then washed in water and dried with blotting paper and observed under oil immersion objective.
For more information and commonly asked questions, visit www.microrao.com/staining.htm
Photo of Gram stained smear showing gram positive cocci and gram negative bacilli
For a detailed description, visit my recent post here or type http://medimicro.blogspot.com/2008/10/gram-stain.html.
For PDf version, click here or type www.microrao.com/micronotes/pg/Gram stain.pdf
Ziehl Neelsen staining
The procedure adopted in our institution is as follows:
The smear is flooded entirely with concentrated carbol fuchsin solution and heated using a spirit lamp from beneath. The heating should be intermittent and should not be intense to boil the solution or dry it completely. Typically, flaming must be stopped once fumes arise and allowed to cool. The solution is then poured off and washed in gentle stream of running tap water. The smear is then covered with few drops of 20% sulfuric acid and allowed to act for 1-2 minutes and then washed in tap water. The process of decolourisation may be repeated until the smear is faintly pink or almost colourless. The smear is then washed in water and counterstained with methylene blue solution and allowed to act for 30 seconds. The slide is then washed in water and dried with blotting paper and observed under oil immersion objective.
A positive sputum sample typically contain pink coloured, rod shaped bacteria that are slightly curved, sometimes branching, sometimes beaded in appearance, present singly or in small clumps against a blue background of pus cells and epithelial cells.
For more information and commonly asked questions, visit www.microrao.com/staining.htm
Photo of acid fast bacilli in sputum smear
Antibiotic susceptibility testing
Within fifteen minutes of standardization of inoculum, a sterile cotton swab is dipped inside the tube and squeezed against the sides of test tube to remove excess of fluid. The moist swab is rubbed on the entire surface of a dry Mueller Hinton agar. This procedure is repeated by streaking two more times, rotating the plate approximately 60o each time so as to obtain an uniform lawn growth. Finally the circular rim of the plate must be swabbed.
The surface of the medium is allowed to dry and filter paper disks containing known antibiotic is placed on the surface and pressed gently to ensure proper contact. Once placed, the discs should not me moved or relocated since diffusion of antibiotic starts instantly. The interdisc distence should be 24 mm to prevent zone overlapping. Ordinarily, no more than 12 discs should be placed on one 150 mm plate or more than 5 discs on a 100 mm plate. Within fifteen minutes of application of discs, the plate is inverted and kept in incubator. The plates are incubated in ambient air or CO2, as required.
After 18-24 hours of incubation, the plates are removed from the incubator and observed for good growth and zones of inhibition around the discs. The diameter of zone of inhibition is measured to the nearest mm and read from the Kirby-Bauer Chart and is reported accordingly as susceptible, moderately susceptible or resistant.
For more information, visit www.microrao.com/micronotes/susceptibility.htm
Bacterial spore
Mechanism of sporulation:
First the DNA replicates and the cell divides asymmetrically. A cytoplasmic membrane septum forms at one end of the cell. A second layer of cytoplasmic membrane then forms around one of the DNA molecules (the one that will become part of the endospore) to form a forespore. Both of these membrane layers then synthesize peptidoglycan in the space between them to form the cortex. Calcium dipocolinate is also incorporated into the forming endospore. A spore coat composed of a keratin-like protein then forms around the cortex. Sometimes an outer membrane composed of lipid and protein and called an exosporium is also formed. Finally, the remainder of the bacterium is degraded and the endospore is released. There is no metabolic activity until the spore is ready to germinate. Single vegetative cell gives rise to a single spore. Sporulation generally takes around 15 hours.
Germination:
Favorable growth conditions signal the process of endospore germination. Germination of a spore results in a break in the spore wall and the outgrowing of a new vegetative cell. The newly formed vegetative cell is capable of growth and reproduction. A single spore upon germination forms a single vegetative cell. Germination occurs in following steps:
Activation: Even in the presence of favorable conditions, the spore will not germinate until its protective spore coat is not damaged. Conditions such as heat, acidity, abrasion or compounds containing free sulphydryl groups activate the spore to germinate.
Initiation: once activated, the spore will germinate provided the environment is suitable. Different signaling effectors exist for different species. Binding of effector stimulates autolytic enzymes that degrade the peptidoglycan of cortex. Water is absorbed and calcium dipicolinate is released.
Outgrowth: once the cortex and outer layers is degraded, a new vegetative cell consisting of spore protoplast and its wall emerges. This is followed by active biosynthetic activity and process terminates with cell division.
The impermeability of the spore coat is thought to be responsible for the endospore's resistance to chemicals. The resistance of endospores is due to a variety of factors:
Calcium-dipicolinate, abundant within the endospore, may stabilize and protect the endospore's DNA. Specialized DNA-binding proteins saturate the endospore's DNA and protect it from heat, drying, chemicals, and radiation. The cortex may osmotically remove water from the interior of the endospore and the dehydration that results is thought to be very important in the endospore's resistance to heat and radiation. DNA repair enzymes contained within the endospore are able to repair damaged DNA during germination.
For more information on anatomy of bacterial cell, visit
www.microrao.com/micronotes/anatomy.pdf