1

SEMINAR ON

INTRODUCTION TO MICROBIOLOGY

Presented by : Naseef.PP Presented on : 17th May 2008 Submitted to : Mr.Dileep.C

2

INTRODUCTION TO MICROBIOLOGY

Microbiology is the study of microorganisms, which are unicellular or cellcluster microscopic organisms. This includes eukaryotes such as fungi and protests, and prokaryotes such as bacteria and certain algae. Viruses, though not strictly classed as living organisms, are also studied. Microbiology is a broad term which includes many branches like virology ( Study of virus), parasitology ( Study of parasites), mycology ( Study of fungi), and others. A person who specializes in the area of microbiology is a microbiologist. SOME IMPORTANT MICROBIOLOGY EVENTS IN THE DEVELOPMENT OF

1608- Jansen develops first useful compound microscope. 1857- Pasture describes fermentation 1882- Koch discovers Mycobacterium tuberculosis 1929- Fleming discovers pencillin 1953- Waston and Crick proposes DNA double helix 1984- HIV isolated and identified by Gallo and Montagnier 2000- Discovered that vibrio cholerea has two chromosomes 2001- Anthrax biotactrorism attack in New York 2002- Infectious poliovirus synthesized from basic chemicals 2003- SARS out break in China

3

Types of microbiology
The field of microbiology can be generally divided into several subdisciplines:

• •

• •

Microbial physiology: The study of how the microbial cell functions biochemically. Includes the study of microbial growth, microbial metabolism and microbial cell structure. Microbial genetics: The study of how genes are organised and regulated in microbes in relation to their cellular functions. Closely related to the field of molecular biology. Medical microbiology: The study of the role of microbes in human illness. Includes the study of microbial pathogenesis and epidemiology and is related to the study of disease pathology and immunology. Veterinary microbiology: The study of the role in microbes in veterinary medicine or animal taxonomy. Environmental microbiology: The study of the function and diversity of microbes in their natural environments. Includes the study of microbial ecology, microbially-mediated nutrient cycling, geomicrobiology, microbial diversity and bioremediation. Characterisation of key bacterial habitats such as the rhizosphere and phyllosphere, soil and groundwater ecosystems, open oceans or extreme environments (extremophiles). Evolutionary microbiology: The study of the evolution of microbes. Includes the study of bacterial systematics and taxonomy. Industrial microbiology: The exploitation of microbes for use in industrial processes. Examples include industrial fermentation and wastewater treatment. Closely linked to the biotechnology industry. This field also includes brewing, an important application of microbiology.

4
• • • •

Aeromicrobiology: The study of airborne microorganisms. Food microbiology: The study of microorganisms causing food spoilage. Pharmaceutical microbiology: the study of microorganisms causing pharmaceutical contamination and spoilage. Oral microbiology: the study of microorganisms of the mouth in particular those causing caries and periodontal disease.

Pharmaceutical Microbiology
Microbes are the part of our lives in more ways than most understand. Microbiology is one of the most applied of all the biological sciences. Microbiology can be divided into two major fields: theoretical and applied. The major fields of applied microbiology are: Medical microbiology, Aquatic microbiology, Aero-microbiology, Agricultural, Industrial microbiology, Exomicrobiology and Geochemical microbiology. Industrial and pharmaceutical microbiology are the most promising areas of applied microbiology. Health care products derived from microbial biosynthesis are antibiotics, hormones, vitamins, and vaccines. Pharmaceutical microbiologists have several tricks to increase the amount of the chosen end product. Use of industrial fermentation processes begin with microbial cells acting as living factories

Industrial microbiology encompasses a broad and complex area of study. It includes the many uses of microorganisms to produce products of economic value and to decompose the wastes of municipalities and industries. In addition it includes the prevention of unwanted growth of microorganisms and the resultant deterioration of natural and man made material.

Industrial microbiology concern itself with the isolation and description of microorganisms from natural environments such as soil or water and with the cultural conditions required for obtaining rapid and massive growth of these organisms in the laboratory and in large scale cultural vessels commonly known as fermentors. MICROSCOPY

5

Microscope has critical importance in microbiology. The science dealing with all aspects of microscope is called as Microscopy. Microscope is an optical instrument used to magnify minute objects or microorganisms, which cannot be seen by naked eye. The objective of microscope are magnify the image, maximize resolution and achieve sufficient contract to differentiate the cellular component. The microscope utilizing as a source of illumination is called as light Microscope. The types of light microscopes are • Bright field Microscope • Dark field Microscope • Flurascence Microscope The Microscope utilizing electron beam as a source of illumination is called as Electron Microscope. This are of two types • Transmission Electron Microscope • Scanning Electron Microscope STERILIZATION Defenition: Sterilization is defined as process by which an article , object, surface, culture medium, specimen or material is freed from all types of living microorganisms including their spores. Methods of sterilization Physical methods 1. Heat

6

a) Dry Heat Red heat Flaming Incineration or burning Hot air oven Infra red conveyor sterilizer and Microwave oven 2. Moist Heat (Steaming) a) Steaming below 100oc or below atmospheric pressure Inspissation b) Steaming at 100oc or at atmospheric pressure Tyndelisation Single exposure for 90 minutes c) Steaming under pressure , above 100oc or above atmospheric pressure Autoclaving

3. Ionizing radiation(cold sterilisation) a) Gamma rays b) Particulate high energy electrons Chemical methods 1. Low Temperature Steam and Fornaldehyde(LTSF) 2. Ethylene oxide gas 3. Formaldehyde gas 4. Betapropiolactone 5. Gas plasma

7

6. Liquid Cheamosterilizers a) Formaline b)Glutraldehyde solution Mechanical methods A) Filttration 1 Diatomaceous earthenware candle filters, e.g.,Mandler filter,Berkfeld filter 2 Unglaced, porcelain filter, e.g., Jenkins filter 3 Asbestose disk filter, e.g., Seits filter 4 Sintered glass filters 5 Membrane filters(Ulta filters) 6 HEPA(High Efficiency Particulate Air) filters

NUTRITIONAL TYPE Phototrophs- Derive energy from sun light. Chemotrophs- Derive energy from oxidation of chemical compounds. Autotrophs (Lithotrophs ) – Can synthesise their own metabolites Hetrotrophs ( Organotrophs) - Cannot synthesise their own metabolites, hence depend upon preformed readymade organic compound.

Bacteria

8

Autotrophs (Lithotrophs )

Hetrotrophs ( Organotrophs)

Photoautotrophs Chemoautotrophs Phototrophs

Photohetrotrophs Chemohetrotrophs Chemotrophs

Bacteria

CULTURE MEDIA Culture media is an artificially prepared mixture of various nutrients in appropriate concentration, for the active growth of microorgsnisms, prepared by considering the biochemical requirements of microbes. There are various types of culture such as Liquid media , solid media, aerobic media, enriched media, differential media etc..
Culture techniques are designed to promote the growth and identify particular bacteria, while restricting the growth of the other bacteria in the sample. Often these techniques are designed for specific specimens; for example, a sputum sample will be treated to identify organisms that cause pneumonia, while stool specimens are cultured on selective media to identify organisms that cause diarrhoea, while preventing growth of non-pathogenic bacteria. Specimens that are normally sterile, such as blood, urine or spinal fluid, are cultured under conditions designed to grow

9

all possible organisms. Once a pathogenic organism has been isolated, it can be further characterised by its morphology, growth patterns such as (aerobic or anaerobic growth, patterns of hemolysis) and staining.

IDENTIFICATION OF MICROORGANISMS
Identification of bacteria in the laboratory is particularly relevant in medicine, where the correct treatment is determined by the bacterial species causing an infection. Consequently, the need to identify human pathogens was a major impetus for the development of techniques to identify bacteria

The microorganisms can be identified as follows

Staining
Stain is defined as an organic compound containing both chromophore and auxochrome groups linked to benzene ring. Chromophore group imparts colour to stain. The group that imparts ability to binding tissue is Auxochrome. The electrolytic dissociation of auxochrome groups helps to bind the stain with the cell. A. Simple Staining 1. Negative or Indirect (Relief) staining In this back ground is stained and the cell remains courless. The bacterial cell possesses slightly negative charge. The acidic stain is used in this staining Eg. Eosin stain. Such stain is not responsible for the staining of the cell because there is repulsion between two similar charges ie, negative charge of acidic stain and negatively charged bacterial cell. Some other stains having molecular size greater than bacterial pores are also useful in such staining. The stain particles remains outside the cell and just stain back ground.

10

2. Monochrome staining The use of single stain to colour bacteria is the monochrome staining. This utilizes the knowledge that bacteria possess slightly negative chrge. The negative charged group of bacterial cell surface produces attraction between basic stains. Eg. Methylene blue. B. Differential staining This differentiate two kinds of microbes. The stains used in this method react differently with various microbes. The differentiation is due to the chemical compositon of organisms. 1. Gram staining This categorizes bacteria into two groups. Gram positive and gram negative. Under oil emersion objective we can observe purple coloured gram positive and pink coloured gram negative bacteria 2. Acid fast staining Several bacteria do not get decolourised even after the application of strong decolouriser like acid alcohol mixture , this property is known as acid fastness. This categorize bacteria in to acid fast and non acid fast. The acid fast bacteria appear pink and non acid fast bacteria appear blue. E.g. mycobacteria or Nocardia, BACTERIAL MOTILITY

11

Some bacteria are motile and move from one place to another by means of flagellae. It is essential to differentiate between true motility and false motility. In true motility , there is a displacement of position ie, the organisms move from one place to another in different directions. The false motility also known as Brownian movement, is an oscillatory vibration , exhibited by both living and non living organisms, due to water convection current. Bacterial motility can be demonstrated by various methods such as wet mount method – hanging drop experiment, microscopic , macroscopic and demonstration of flagella. Motile organisms – Eschechia coli, Entero bacteria Non Motile organisms – Klebsialla, shigella GENARAL METHODS OF CLASSIFYING BACTERIA
Classification seeks to describe the diversity of bacterial species by naming and grouping organisms based on similarities. Bacteria can be classified on the basis of cell structure, cellular metabolism or on differences in cell components such as DNA, fatty acids, pigments, antigens and quinones. While these schemes allowed the identification and classification of bacterial strains, it was unclear whether these differences represented variation between distinct species or between strains of the same species. This uncertainty was due to the lack of distinctive structures in most bacteria, as well as lateral gene transfer between unrelated species. Due to lateral gene transfer, some closely related bacteria can have very different morphologies and metabolisms. To overcome this uncertainty, modern bacterial classification emphasizes molecular systematics, using genetic techniques such as guanine cytosine ratio determination, genome-genome hybridization, as well as sequencing genes that have not undergone extensive lateral gene transfer, such as the rRNA gene. The term "bacteria" was traditionally applied to all microscopic, single-celled prokaryotes. However, molecular systematics showed prokaryotic life to consist of two separate domains, originally called Eubacteria and Archaebacteria, but now called Bacteria and Archaea that evolved independently from an ancient common ancestor. The archaea and eukaryotes are more closely-related to each other than

12

either is to the bacteria. These two domains, along with Eukarya, are the basis of the three-domain system, which is currently the most widely used classification system in microbiolology. However, due to the relatively recent introduction of molecular systematics and a rapid increase in the number of genome sequences that are available, bacterial classification remains a changing and expanding field. For example, a few biologists argue that the Archaea and Eukaryotes evolved from Gram-positive bacteria.

Two methods are used for arranging bacteria 1. The intuitive method A microbiologist who is thoroughly familiar with the properties of the organisms he or she has been studying for several years may decide that the organisms represents one or more species or genera. 2. Numerical taxonomy A scientist may determine many characteristics ( usually 100 – 200) for each strain studied , giving each characteristics equal weight. Then using a computer he or she calculate the percentage similarities of each strain to every other strain. NS %S = NS+ND NS – Number of characteristics that are same ND - Number of characteristics that are different. Those streams having a high % S to each other are placed into groups

13

Gram-negative bacteria The proteobacteria are a major group of Gram-negative bacteria. Other notable groups of Gram-negative bacteria include the cyanobacteria, spirochaetes, green sulfur and green non-sulfur bacteria. Gram negative bacteria are those that are decolourised completely and take up saffranin counter stain, appearing red or pink. These also include many medically relevant Gram-negative cocci, bacilli and many bacteria associated with nosocomial infections. Gram-positive bacteria In the original bacterial phyla, the Gram-positive forms made up the phylum Firmicutes, a name now used for the largest group. It includes many wellknown genera such as Bacillus, Listeria, Staphylococcus, Streptococcus, Enterococcus, and Clostridium. It has also been expanded to include the Mollicutes, bacteria like Mycoplasma that lack cell walls and so cannot be stained by Gram, but are derived from such forms. Gram positive bacteria are those that take up and retain violet dye and resist decolourisation, appearing violet or purple.

14

The scope of microbiology Microbiology is not a single subject. It has many areas of specialization. However it is a science based on the use of pure cultures - looking at one sort of organism at a time. Many of the techniques developed by microbiologists are now used in molecular and cell biology to provide the basis for studying higher organisms. Microorganisms also play a central role in recombinant DNA technology and act as the agents for genetic modification. Applying knowledge gained from these techniques can lead to many improvements in the quality of our lives. In this way microbiology makes an important contribution to biotechnology, an area of science that applies industrial techniques to biological processes. PRODUCTS OF MICROBIAL ACTIVITY HAVING COMMERCIAL IMPORTANCE 1. Antibiotics – streptomycin, pencillin, tetracycline, erythromycin 2. Organic solvents – acetone, butanol, ethanol, 3. Gases – CO2 , hydrogen 4. Beverages – wine, beer, and distilled 5. Foods – cheese, picks, yeast 6. Organic acids –lactic acid, citric acid 7. Amino acids – L-glutaric acid, L-lysine 8. Vitamins – B12 , vitamin A 9. Enzymes – protease, pectinases The scope of microbiology may extent in Applied microbiology and in Basic microbiology. The applied microbiology includes disease related such as chemotherapy, infection control by preventing the spread of disease , environmental and industrial microbiology. Industrial microbiology includes Genetic engineering, pharmaceutical production of drugs and in food & Beverage technology. The basic microbiology includes the microbe taxonomy and various field of microorganisms such as virology, mycology etc.

15 Industrial microbiology

a. Microbes have played important roles in manufacturing products for as long as there has been history. b. Microorganisms are used to: i. ferment useful chemicals (ethanol, acetone, etc.) ii. produce certain food stuffs (wine, cheese, yogurt, bread, half sour pickles, etc.) iii. produce of recombinant products (recombinant insulin, human growth hormone, etc.) iv. destroy wastes (sewage, oil spills, bioremediation)
Medical microbiology

A Microbes both cause and prevent disease. B Microbes produce antibiotics used to treat disease. C The single most important achievement of modern medicine is the ability to treat or prevent microbial disease. D Most of this course will consider the physiology of microbes and their role in disease.
Pathogen

A A microorganism is considered to be a pathogen or pathogenic if it is capable of producing disease. B Though only a minority of microorganisms are pathogenic, practical knowledge of microbes is necessary for their treatment so is highly relevant to medicine and related health sciences.

REFERENCES 1 www.wikipedia.com

16

2 www.ask.com 3 www.microbiologyprocedure.com 4 Ruiz-Bravo, A. and Jiménez-Valera, M. Ars Pharm.37;(2);171-181, (1996) 5 Supplemental Lecture (98/03/28 update) by Stephen T. Abedon 7 Foundation in microbiology, Dr. A.B Chaudhari 6 Microbiology- Michael J. Pelczer,JR 8 Prescortt, Harley,and Klein’s Microbiology 9 Text book of microbiology, Rajeshware Reddy