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Introduction To General Microbiology

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Title: Introduction To General Microbiology

1
Introduction To General Microbiology

 Prof. Dr. Asem Shehabi

 Faculty of Medicine
 University of Jordan

The Microbial World

The microbial world is composed of commensally

and pathogenic Microbes/ Microorganisms..

Bacteria, Fungi (Yeast/ Moulds), Algae, Protozoa/

Parasites and viruses.

Microbiology is concerned with the study of these

microbes.. Mostly are beneficial.. Few species

cause harmful effects ..disease in human

animals.

Microorganisms are unicellular cell.. too small

to be seen with the naked eye.. recognized by

light microscope .. Bacteria, fungi Parasites

..their sizes above gt 0.1 um

Most microbes capable of grow existence as

single organism or together with others .. Widely

distributed in Human, Animal, Nature.

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Microbiology

 Viruses sizes lt 0.01um Composed of only DNA or

RNA.. grow only in living cells/tissue culture..
are non independent cellular entities..cant be
considered true microorganisms..Their presence
structures can be seen only with electron
microscope.
 Microbiology has many areas of specialization
including Bacteriology, Mycology (fungi),
Virology, Medical microbiology, Immunology, Food
microbiology, Biotechnology, Microbial genetics
..Industry.. Agriculture Veterinary.

4
Classification of Microorganisms


 Two fundamentally different types of cells are
classified in the microbial world, Prokaryotic ..
Eukaryotic cells.
 Eukaryotic cells have a "true" nucleus..
Prokaryotic cells have a naked nucleus.. composed
of a single DNA Chromosome.. not enclosed within
a nuclear membrane.
 The shape of Prokaryotic cells.. Bacterial, Fungi
,Parasites cells are of fundamental importance
in the classification and identification of these
microbes in Labs.

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Bacteria

 Bacteria are unicellular microorganisms.. Size

 (0.2umDiameter, 0.2-10um Length) having a
variety of shapes ..Growth patterns metabolic
characteristics allowing their classification.
 Major bacteria cell shapes are arranged
Coccus/cocci, Bacillus/bacilli or Rods,
Coccobacilli, Spiral forms- spirochetes, Vibrios
 Individual cells may be arranged in pairs or
clusters or chains.. Their morphologies are
useful for the identification classification of
bacterial Genera and Species.. colored by
Gram-stain or other stains (Fig-1)

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Fig-1 Gram-Negative/positive
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Figure -2 Bacteria Cell structure
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Bacterial Cell structures-1

 Cell wall structures A rigid cell wall, composed

of many peptidoglycan layers .. outer membrane, A
periplasmic space, a cytoplasmic membrane lacking
sterols, Cytoplasma ..70S ribosomes, mesomes,
storage granules -Lipids, glycogen,
 polysaccharides, sulfar, phosphate .. Others
storage compounds.
 Bacterial genome.. One single supper coiled DNA
chromosome, plasmids(gt1).
 Flagella Organs of motility, composed of
flagellins (polymer proteins) long filament..
length up to 20 um (Figure 2).. Attachment..
Nutrition..Single polar flagellum
(monotrichous).. Several polar flagella at one,
each end of the cell or covering the entire cell
surface (peritrichious).. antigenic determinants
(H-antigen)..observed during bacterial infection.

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Bacterial Cell structures-2

 Fibmriae.. Pili Small surface appendages

(protein).. Few numbers Pili.. Sex function
/Large Numbers fimbriae..specific functions ..
Attachment/Adhesion to host epithelial
cells/colonization antigenic determinants.
 Capsules surface layer of cell wall.. a slime
layer composed mostly of high molecular weight
polysaccharides.. provide resistance to
phagocytosis.. avoid the killing effects of
lysosomal enzymes, and serve as antigenic
determinants.. (K-antigen).. Major virulence
factor in certain bacteria
 Virulence factor.. Any bacterial part/product
Associated with pathogenic potential.. causing
human/animal infection/disease.

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Bacterial Cell wall Structures-1

 Bacterial cell wall contains a special polymer

called Peptidoglycan. Its basic structure is a
carbohydrate backbone of alternating units of
N-acetyl glucosamine and N-acetyl muramic acid.
 These are cross-linked with oligopeptides..
contain both D- and L-amino acids.
 Teichoic acid-Lipoteichoic acids found only in
Gram-positive bacteria.
 Lipopolysaccharides Lipopolysaccharides (LPS)
found only in Gram-negative bacteria.

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Cell wall Gram-positive bacteria-3
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Cell Wall Gram-negative bacteria-4
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Bacterial Cell wall Structures-2

 LPS structures are composed of lipid A, which

binds to the outer membrane.. Endotoxic portion
 of the molecule.. Causing Toxic Shock.. High
Fever, Sepsis
 The polysaccharide moiety appears on the cell
surface, serving as an antigenic determinant O
antigen- Host cells develop during bacterial
Infection..Anti-O AB
 Cell wall is the basis for classification of
bacteria into Gram-positive Gram-negative by
Gram-stain
 Cell membrane A phosolipid bilayer responsible
for transport of ions, nutrients and waste across
the membrane.. Control the cell plasma contents

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Gram-Stain

 A- Gram-positive bacteria have a thick layer of

peptidoglycan, Many sheets.. external to the
cytoplasmic membrane.. Lipoteichoic acids..
stained Blue.. Staphyloccocus, Streptocooci,
Bacillus..Protoplasts..L-form..Lysozyme
effect..Loss Most Cell wall, Burst Lysis
 B- Gram-negative bacteria contain
lipopolysaccharide (LPS) attached to the outer
membrane... source of the O-antigen and endotoxin
reaction.. Stained Purpel/Red.. Enteric bacteria
group.. Esch. coli, Klebsiella, Salmmonella
Pseudomonas, Spheroplasts



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Spore-Forming Bacteria

 ENDOSPORE FORMATION The process of sporulation

begins when vegetative (actively growing cells)
exhaust their source of nutrients .. begin of
forming endospores.. Common in nature (Figure 4).
 Spore forming Bacteria are very resistant to
lysozyme, heat, radiation, drying and can remain
dormant for hundreds of years in nature.. Once
conditions are again favorable for growth, the
spores can germinate and return to the vegetative
state.
 Aerobic Bacillus group Anaerobic Clostridium..
develop Endospore formation.. Both are widely
distributed in nature ..intestinal -human and
animals.

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Bacterial Spore -Fig-5
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Growth Nutrition-1

 Requirements for bacterial growth oxygen, water,

pH, temperature, source of carbon, nitrogen (
organic compounds), inorganic salts.. Na, K, S,
P, Ca, Mg, Cl, Fe, vitamins, etc.
 Obligate Aerobic bacteria ..M. tuberculosis,
P.aeruginosa grow using respiration.. oxidation..
 recipient Oxygen.. Aerobic bacteria encounter the
oxygen damage during their growth by producing
oxidizing enzymes
 Peroxidase Oxidize H2O2 into 2H2ONAD.
 Superoxidase dismutase Reduce O2- into H2O2 O2
..
 Catalase Reduce H2O2 into 2H2OO2.

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Growth Nutrition-2

 Certain Pathogens grow with reduced level of

oxygen.. Microaerophilic bacteria..Neisseria
spp.
 facultative anaerobes.. prefer growing in the
presence of oxygen, but can continue to grow
without it.. Most human pathogens normal
flora.. Staphylococci, streptococci, E.coli
 Obligate Anaerobic bacteria grow by absence of
oxygen.. use recipient inorganic molecule..
Fermentation.. Mostly found in intestinal tract
(95-99), Mouth Vagina(90)
 Examples Anaerobes Gram-ve Bacteriodes
fragillis, Gve Clostridia, Gramve Cocci

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Growth Nutrition-3

 Bacteria classified by the source of their energy

oxidation-reduction process into two groups
 Heterotrophs derive energy from breaking down
complex organic compounds.. protein, sugar,
fats.. human tissues.. All commensals-pathogens
 Autotrophs fix carbon dioxide to make their own
food source.. using light energy
photoautotrophic, or oxidation of nitrogen,
sulfur, other elements chemoautotrophic.. sulfur
nitrogen fixing bacteria.. Environment.
 Saprophytic bacteria/ Nonpathogenic.. take energy
by fermentation/respiration.. found in nature..
in decaying material.. soil, water..vegetations..c
irculation of minerals.

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 4/
 Culture Media Nutrients (carbohydrates
proteins, blood, minerals) Source.. Water..Broth
medium, Solid medium/ Blood agar, Petri
dishes/Plate, Growth/Culture (Fig 5)
 Neutrophilic bacteria.. Grow best pH (7-7.2) Most
human-animal commensales pathogens
 Acidophilic Bacteria lt 5 pH.. Lactobacilli
 Mesophilic Bacteria (20-40C)..Most human
commensal pathogens.
 Psychrophilic bacteria(lt10C), Thermophiles
bacteria (gt 60C)..Common in hot spring water
 Counting bacteria growth Plate counts,
Turbidity, Dry weight using solid culture agar
22
Bacterial growth-1

 Bacterial growth is the division of one bacterial

cell into 2 identical daughter cells..4,8.16..
binary fission..Generation time ( 15-25 min),
most human commensal pathogens.. Each produce
one colony 103 -109 cells ( Fig-4).
 Bacterial Strain originated from a single cell.
 Baterial Growth Curve 4 phases of visible
growth...Lag, Log, Stationary, death/ decline.
 Measurement of bacterial growth followed by
 A) Growth/enumeration of cells by direct cell
counting in nutrient broth.. microscopic or
counting viable cells/ colony forming unit..
Plate counts/ Electronic counting..using solid
culture media..nutrient agar

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Binary fission Bacteria (Fig-6)
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Bacterial Growth MacConkey agar Tube Broth
(Fig-7)
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2/Measurement of Growth

 B) Indirect counting of growth in fluid medium..

most probable number by measuring turbidity, wet
or dry weight.. G/ml.. Important in study
research to detect antibiotics treatment of
infection.
 Types of culture media
 General culture media Nutrient agar, blood agar,
chocolate agar..growth of most human pathogens..
Gram-ve Gram-ve bacteria.
 Selective differential media..MacConkey agar
 Bile salts Lactoseneutral red dye ..Gram-ve
bacteria, E.coli, other enteric bacteria
 Selective media S-S agar .. For Isolation of
Salmonella, Shigella , V.colerae from stool
specimens.

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Bacteria Growth Curve (Fig-8)