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FUNDAMENTAL FEATURES

OF MICROBIOLOGY
Chapter 2
Introduction

Classification of microorganisms
Bacteria, Fungi
Protozoa , Viruses
Chlamydia, Rickettsia
Mycoplasmas
Ureaplasmas
Viruses

DNA or RNA Lipid


surrounded envelopes
by proteins

Glycoproteins
Viruses

Intracellular parasites

Helical, linear, or spherical

20-400 nm in size

Single or double stranded DNA or RNA


Viroids or Virusoids

Single-stranded RNA

Plant pathogens

Maybe origin of Hepatitis D


virus
Prions

No nucleic acid

Atypical mammalian protein

Creutzfeldt-Jacob syndrome

Bovine spongiform encephalopathy


Classification of Microorganisms

Fungi
Prokar Bacteria Eukar- Protozoa
yotes Archeae yotes Algae
Differences of Prokaryotes and
Eukaryotes
Prokaryotes Eukaryotes
No true nucleus True nucleus
Single stranded Chromosome
chromosome not separated from
separated from cytoplasm
cytoplasm
Haploid Diploid
Archaea

Extreme
thermophiles

Extreme halophiles

Derive energy from sulfur


or iron and production of
methane
Eubacteria
Cocci
Shape Bacilli
Spirilla

Gram Gram positive


reaction Gram negative

Pathogens
Action Saprophytes
Bacillus
megaterium
Micrococcus
luteus
Obligate Intracellular Parasites

Rikettsia

Chlamydia
Bacteria of Pharmaceutical Importance

Possess cell
wall

Facultative Mesophiles
anaerobes

Strict Strict
anaerobes aerobes
Fungi

Non-photosynthesizing
plants

Yeasts

Moulds or molds
Yeasts

5-10 m

Unicellular organisms

Divide by binary fission or budding


Saccharomyces
cerevisiae
Mould or Mold

Mycelium (mass of filaments)

Hyphae (mass of threads)


Toadstool and mushroom
Mucor
plumbeus
Streptomyces
rimosus
Aspergillus
niger
Protozoa

Free-living motile organisms


Parasites of plants and Soil and
animals including humans water

Malaria Amoebic Pathogenic


dysentery
Naming of Organisms

Binomial Nomenclature

Genus species

Upper case Lower case


Microbial Metabolism

Chemoheterotrophs
Obtain C, N, and energy
by breaking down
organic compounds
Microbial Metabolism

Oxidation Reduction
accept loss of
electrons electron
Microbial Metabolism

Glycolysis Fermentation
Aerobic respiration Sugars
Anaerobic Ethanol, CO2, acid,
respiration alcohol
Microbial Metabolism

Mineral Sugar Glycerol


salts

Lactic Pyruvic Carbohy


acid acid drates

Proteins
Action of Antibiotic

Exploit on difference of metabolism

Toxicity

Interfere with reaction unique to


bacteria
Microbial Metabolic Products

Ethanol
Primary Organic acids
metabolites

Antibiotics
Secondary Enzymes
metabolite Toxins
Carbohydrates
Secondary Metabolites

Antibiotics
Amylase
Enzymes Biological washing powders

Botox
Toxins Clostridium botulinum

Carbohy- Dextran
drates Plasma expander
Microbial Cultivation

Culture

Simple Cell In living


technique culture animal
Culture Media

anything that possess the


nutritional and environmental
needs necessary for the
growth and multiplication of
microorganisms
Classification of Culture Media

Function Form

Consistency Composition
Types of CM Based on Function

Basal Enriched

Differential Selective
Basal/Basic/General/Simple/Ordinary

Allows growth of a lot of


microorganisms

Beef extract, yeast extract,


peptone

Nutrient agar, nutrient broth


Nutrient agar Nutrient broth
Enriched Media

Basal medium + enrichment


substance
Blood, serum, ascitic fluid,
milk

BAP, CAP
BAP
CAP
Differential Media

Indicator or dye

Differentiates groups of
microorganisms

EMB, McConkey, HEA,


MSA, TCBS
McConkey Agar
EMB
HEA
TCBS
MSA
Selective Media

Inhibitory substances

Allow growth of some


organisms but suppress others

SSA, BSA, Thayer-Martin


SSA
BSA
Types of CM Based on Form

Butt or deep
Slant
Tubed Butt-slant
Broth

Plated
Butt or Deep
Butt-Slant
Broth
Types of CM Based on Consistency

Solid Semi- Liquid


solid
Types of CM Based on Composition

Synthetic

Non-synthetic
Cultivation Methods

Binary fission

Generation time 25-30 minutes

Nutrients are exhausted

Toxic metabolites accumulate


Cultivation Methods

Liquid Turbidity
media
Solid Colony
media
Anaerobic Cultivation

Catalyst Palladium-coated pellet or wire

H2O + sodium Hydrogen Oxygen


borohydride
Anaerobic Cultivation

Ascorbic Redox
acid indicator

Resazurin
Anaerobic Cultivation

Gas pack Candle Jar


Yeast Colony

Larger than bacteria


Colored
Pigmented central area
Non-pigmented periphery
Planktonic and Sessile Growth

Planktonic
Freely suspended cells in liquid

Sessile
Biofilm or microcolony mode of
growth
Enumeration of Microorganisms

Measuring the Evaluating the


levels of effects of an
microbial antimicrobial
contamination in chemical or
a raw material decontamination
or manufactured process
medicine
Enumeration of Microorganisms

Using
microorganisms Assessing the
in the nutrient
manufacture capability of a
of therapeutic growth medium
agents
Microbial Counting

Total count

Viable count

Total viable count


Methods of Viable Counts

Pour plate

Surface spread or surface drop(Miles Mitra)

Membrane filter

Most Probable Number


Serial Dilution
Pour Plate and Spread Plate
Methods
Membrane Filter Method
Membrane Filter Method
Most Probable Number
Methods of Total Counts

Direct microscopic counting

Turbidity method

Dry weight determination


Nitrogen, protein, or nucleic acid
determination
Direct Microscopic Counting
Turbidity Method
Limitations of Traditional Methods

Labor intensive and hard to


automate

Time consuming

Require large volumes of CM, petri


dishes, and incubator space
Rapid Methods

Epifluorescence
ATP luciferin
Impedance
Manometric gas
consumption or production
Epifluorescence Microscope
Epifluorescence
Manometric Method
Microbial Genetics of Bacteria

Genome
Chromosomes (double-
stranded DNA)
Plasmids
Bacteriophage
Bacterial Chromosome

1 mm or 1000-3000
more genes

Multiple
copies
Plasmids

0.1- Linear Survival


1.0%

Multiple Two or
copies more
Plasmids

Replicate
independently Pili

Pass from 1
cell to another Conjugative

Host Cross between


specificity species or
genera
Microbial Genetics of Eukaryotes
I or more Asexual
copies of division -
linear mitosis
chromosomes

Sexual
division -
meiosis
Genetic Variation and Gene
Expression
Genotype
Phenotypic adaptation

Genotypic adaptation

Mutation or conjugation
Pharmaceutical Importance of the
Major Categories of Microorganisms

Viruses Prions Bacteria

Fungi Protozoa
Viruses

Pathogenic Incapable of growing


potential on medicines

Not susceptible to Do not cause


antibiotics spoilage

Hazard category Easy to destroy


Prions

Cause incurable, fatal diseases

Resistance to lethal agents

Withstand sterilizing conditions


Bacteria

Pathogens

Resistant to antibiotic and


biocides
MRSA, VSE, multiply
resistant M. tuberculosis
Bacteria

Antibiotics
Sources (streptomycetes)
of Steroids, enzymes, CHO

Survive dry conditions


Agent of Adverse environment
spoilage (heat, radiation, toxic
chemicals)
Fungi

Contaminant

Opportunistic
pathogens
Protozoa

Potential pathogens

Veterinary medicine

Tropical areas
Preservation of Microorganisms

-80oC or lower in refrigerator


Freezing -196oC in liquid nitrogen
Freeze-drying (lyophilization)

Cryopro- Glycerol/dimethylsulphoxide
tectant 10% (v/v) in liquid medium