Friday, November 2, 2012

Standardizing bacterial population in a culture

There are several methods of determining bacterial cells in a culture 1) Direct counts are reliable ways to distinguish biomass of bacteria in natural waters. The usual method is to concentrate the bacteria onto membrane filters, stain them with acridine orange and examined under epifluorescence microscope. 2) Indirect viable count or plate counts involve plating out spreading a sample of culture onto nutrient agar surface. Each colony that can be counted is called colony forming units and the number of cfu is related to the viable number of bacteria in the sample 3) Most probable number (MPN) technique is an important technique in estimating microbial populations in soils, waters, and agricultural products. Many soils are heterogeneous, therefore exact cell numbers of an individual organism can be impossible to determine. The MPN technique is used to estimate microbial population sizes in situations like this. The technique does not rely on quantitative assessment of individual cells, instead it relies on specific qualitative attributes of the microorganism being counted. The important aspect of MPN methodology is the ability to estimate a microbial population size based on a process-related attribute. 4) Turbidity measurements employ a variety of instruments to determine the amount of light scattered by a suspension of cells. Particulate objects such as bacteria scatter light in proportion to their numbers. The turbidity or optical density of a suspension of cells is directly related to cell mass or cell number, after construction and calibration of a standard curve. The method is simple and nondestructive, but the sensitivity is limited to about 107 cells per ml for most bacteria.
For standardizing bacterial population in a culture, make at least three daily transfers in nutrient broth (10µl). Spread a loopful of the sample onto an agar plate with 0.5mm diameter loop and incubate it for 18h. From the agar culture transfer a 1.0 cm x 0.5cm of the culture onto 10mL nutrient broth.

Saturday, July 21, 2012

Monday, February 6, 2012

Friday, August 26, 2011

Saturday, August 13, 2011

Saturday, July 30, 2011

Lectures in Basic Microbiology

Lecture 1 Introduction to Microbiology



Lecture 2 Taxonomy, Phylogeny and Evolution



Lecture 3 Cell morphology and ultrastructures, Classification and Identification



Lecture 4 Biochemical Characteristics and other ID



Lecture 5 Physiological Requirements



Lecture 6 Bacterial Growth and Metabolism



Lecture 7. Microbial Genetics



Lecture 8 Regulatory Mechanisms


Lecture 9 Modified forms of Bacteria


Lecture 10 Mycota


Lecture 11 Protozoa


Lecture 12 Viruses


Lecture 13 Infection and Contagion


Lecture 14 Chemotherapy


Lecture 15 Bioassay