Dr.

Taghreed Khudhur Mohammed

Bacterial Classification, Structure and Function

Introduction
The purpose of this lecture is to introduce you to terminology used in
microbiology. The lecture will:
1. Cover different classification schemes for grouping bacteria, especially the use of
the Gram stain.
2. Describe the different types of bacteria.
3. Discuss bacterial structure and the function of the different bacterial components.
4. Discuss the distinguishing characteristics of Gram positive and Gram negative
bacteria.

Classification Systems
The classification of bacteria serves a variety of different functions. Because of
this variety, bacteria may be grouped using many different typing schemes. The
critical feature for all these classification systems is an organism identified by one
individual (scientist, clinician, epidemiologist), is recognized as the same organism
by another individual. At present the typing schemes used by clinicians and clinical
microbiologists rely on phenotypic typing schemes including:
 Bacterial morphology and staining properties of the organism.
 O2 growth requirements of the species combined with a variety of biochemical
tests.
 For clinicians, the environmental reservoir of the organism, the vectors and
means of transmission of the pathogen are also of great importance.
 Scientists interested in the evolution of microorganisms are more interested in
taxonomic techniques that allow for the comparison of highly conserved genes
among different species. As a result of these comparisons a phylogenetic tree
Dr. Taghreed Khudhur Mohammed

can be developed that displays the degree of relatedness of different organisms.

A relatively new application of this technology has been the recognition and
characterization of non- cultivatable pathogens and the diseases that they
cause.

A. Phenotypic classification systems:

Gram stain and bacterial morphology: Discovered by H.C. Gram in 1884 it
remains an important and useful technique to this day. It allows a large
proportion of clinically important bacteria to be classified as either Gram
positive or negative based on their morphology and differential staining
properties.
Slides are sequentially stained with:
 Crystal violet.
 Iodine.
 Alcohol.
 Counter-stained with safranin.

 Gram positive bacteria stain blue-purple.

 Gram negative bacteria stain red.

The difference between the two groups is believed to be due to a much larger
peptidoglycan (cell wall) in Gram positives. As a result the iodine and crystal
violet precipitate in the thickened cell wall and are not eluted by alcohol in
contrast with the Gram negatives where the crystal violet is readily eluted from
the bacteria. As a result bacteria can be distinguished based on their morphology
and staining properties. Some bacteria such as Mycobacteria (the cause of
tuberculosis) are not reliably stained due to the large lipid content of the
Dr. Taghreed Khudhur Mohammed

peptidoglycan. Alternative staining techniques (Kinyoun or acid fast stain) are

therefore used that take advantage of the resistance to destaining after lengthier
initial staining.

B. Growth Requirements: Microorganisms can be grouped on the basis of their

need for.
C. Biochemical reactions: series of biochemical studies use to identify the
bacterial species.
D. Serologic systems: Selected antisera can be used to classify different bacterial
species. This may be based on either carbohydrate or protein antigens from the
bacterial cell wall or the capsular polysaccharide. (Group A streptococcal M
proteins or O and H polysaccharide antigens of Salmonella).
E. Environmental Reservoirs: When considering likely pathogens it is also
important to know which of the different species are found in different
locations. Environmental reservoirs are generally divided into:
 Endogenous (i.e., on or within the human body).
 Exogenous (somewhere in the environment). When considering the
likely cause of an infection the likely source of the infection is important
in your differential diagnosis. For example, an anaerobic organism
resident in the large bowel is the likely cause of an abdominal abscess
that develops after large bowel surgery. A skin rash developing in a hiker
with a history of multiple tick bites is more likely to be Borrelia, the
agent of Lyme disease. An outbreak of food poisoning traced to imported
unpasteurized cheese might be due to Listeria. Endogenous reservoirs
account for a large proportion of human infections. Many parts of the
body have their own normal flora. S. epidermidis is found on the skin.
Dr. Taghreed Khudhur Mohammed

Viridans streptococci are a part of the normal oropharyngeal flora and S.

aureus is a commensal of the anterior nares.
F. Genotypic systems: Universal Phylogenetic Tree: Woese has developed a
“universal phylogenetic tree” for all living organisms that establishes a tripartite
division of all living organisms– bacteria, archaea and eucarya. His work is
based on a comparison of 16s ribosomal RNA sequences. These sequences are
highly conserved and undergo change at a slow, gradual and consistent rate.
They are therefore useful for making comparisons among the different living
organisms. Ribosomal RNA (rRNA) sequence analysis: This has emerged as
a major method for classification. It has been used (as described above) to
establish a phylogenetic tree. In addition, it is now also used to rapidly diagnose
the pathogen responsible for an infection, to help select appropriate therapy and
to identify noncultivatable microorganisms.
G. Molecular subtyping: Sometimes it is necessary to determine whether strains
from the same species are the same or different. For example, if there is an
outbreak of infections that appear due to the same bacterial species, the hospital
epidemiologist will want to know if all of the infections are due to the same
strain. Clues can be obtained by examining the biochemical studies or the
antibiotic susceptibility profile, but a more reliable method is by molecular
analysis. Pulsed Field Gel Electrophoresis (PFGE) is the most frequently used
molecular technique. Chromosomal DNA is digested with a restriction enzyme
that makes relatively infrequent cuts in the DNA and as a result creates large
DNA fragments. The DNA fragments from the different strains are then run on
a gel and compared.
Dr. Taghreed Khudhur Mohammed

Other Classification of bacteria

A. on the basis of mode of nutrition.

B. on the basis of optimum temperature requirement for growth.

C. on the basis of optimal pH for growth.

D. on the basis of salt concentration.

E. on the basis of gaseous requirement.

F. on the basis of morphology.

H. on the basis of flagella.

I. on the basis of spore.

A] Classification of bacteria on the basis of mode of nutrition

1. Phototrophs: Those bacteria which gain energy from light.

 Phototrophs are further divided into two groups on the basis of source of
electron.
 Photolithotrophs: these bacteria gain energy from light and uses reduced
inorganic compounds such as H2S as electron source. Eg. Chromatium okenii
 Photoorganotrophs: these bacteria gain energy from light and uses organic
compounds such as succinate as electron source.

2. Chemotrophs: Those bacteria gain energy from chemical compounds

 They cannot carry out photosynthesis

 Chemotrophs are further divided into two groups on the basis of source of
electron.
Dr. Taghreed Khudhur Mohammed

 Chemolithotrophs: they gain energy from oxidation of chemical compound

and reduces inorganic compounds such as NH3 as electron source.
Eg. Nitrosomonas
 Chemoorganotrophs: they gain energy from chemical compounds and uses
organic compound such as glucose and amino acids as source of electron.
eg. Pseudomonas pseudoflava

3. Autotrophs: Those bacteria which uses carbon dioxide as sole source of carbon to
prepare its own food.

 Autotrophs are divide into two types on the basis of energy utilized to assimilate
carbon dioxide. Ie. Photoautotrophs and chemoautotrophs.
 Photoautotrops: they utilized light to assimilate CO2. They are further
divided into two group on the basis of electron sources. Ie. Photolithotropic
autotrops and Photoorganotropic autotrops
 Chemoautotrophs: they utilize chemical energy for assimilation of CO2

4. Heterotrophs:

 Those bacteria which uses organic compound as carbon source.

 They lack the ability to fix CO2.
 Most of the human pathogenic bacteria are heterotropic in nature.
 Some heterotrophs are simple, because they have simple nutritional requirement.
However there are some bacteria that require special nutrients for their growth;
known as fastidious heterotrophs.

B] Classification of bacteria on the basis of optimum temperature of growth

1. Psychrophiles:
 Bacteria that can grow at 0°C or below but the optimum temperature of growth
is 15 °C or below and maximum temperature is 20°C are called psychrophilic
bacteria.
 Psychrophiles have polyunsaturated fatty acids in their cell membrane which
gives fluid nature to the cell membrane even at lower temperature.
Dr. Taghreed Khudhur Mohammed

 Examples: Vibrio psychroerythrus, vibrio marinus, Polaromonas vaculata,

Psychroflexus

2. Psychrotrophs (facultative psychrophiles):

 Those bacteria that can grow even at 0°C but optimum temperature for growth
is (20-30)°C

3. Mesophiles:
 Those bacteria that can grow best between (25-40) °C but optimum temperature
for growth is 37°C
 Most of the human pathogens are mesophilic in nature.
 Examples: E. coli, Salmonella, Klebsiella, Staphylococci.

4. Thermophiles:

 Those bacteria that can best grow above 45°C.

 Thermophiles capable of growing in mesophilic range are called facultative
thermophiles.
 True thermophiles are called as Stenothermophiles, they are obligate
thermophiles,
 Thermophiles contains saturated fatty acids in their cell membrane so their cell
membrane does not become too fluid even at higher temperature.
 Examples: Streptococcus thermophiles, Bacillus stearothermophilus

 5. Hyperthermophiles:

 Those bacteria that have optimum temperature of growth above 80°C.

 Mostly Archeobacteria are hyperthermophiles.
 Monolayer cell membrane of Archeobacteria is more resistant to heat and they
adopt to grow in higher temperature.
 Examples: Thermodesulfobacterium
Dr. Taghreed Khudhur Mohammed

C] Classification of bacteria on the basis of optimum pH of growth

1. Acidophiles:

 Those bacteria that grow best at acidic pH

 The cytoplasm of these bacteria are acidic in nature.
 Some acidophiles are thermophilic in nature, such bacteria are called
Thermoacidophiles.
 Examples: Thiobacillus

2. Alkaliphiles:

 Those bacteria that grow best at alkaline pH

 Example: Vibrio cholerae: oprimum pH of growth is 8.2
3. Neutrophils:

 Those bacteria that grow best at neutral pH (6.5-7.5)

 Most of the bacteria grow at neutral pH
 Example: E. coli

D] Classification of bacteria on the basis of salt requirement

1. Halophiles:

 Those bacteria that require high concentration of NaCl for growth.

 Cell membrane of halophilic bacteria is made up of glycoprotein with high
content of negatively (-Ve) charged glutamic acid and aspartic acids. So high
concentration of Na+ ion concentration is required to shield the –ve charge.
 Example: Archeobacteria, Halobacterium, Halococcus

2. Halotolerant: Most of the bacteria do not require NaCl but can tolerate low
concentration of NaCl in growth media are called halotolerant
Dr. Taghreed Khudhur Mohammed

E] Classification of bacteria on the basis of gaseous requirement

1. Obligate aerobes:

 Those bacteria that require oxygen and cannot grow in the absence of O2.
 These bacteria carryout only oxidative type of metabolism.
 Examples; Mycobacterium, Bacillus

2. Facultative anaerobes:

 Those bacteria that do not require O2 but can use it if available.

 Growth of these bacteria become batter in presence of O2
 These bacteria carryout both oxidative and fermentative type of metabolism
 Examples: E. coli, Klebsiella, Salmonella

3. Aerotolerant anaerobes:

 Those bacteria do not require O2 for growth but can tolerate the presence of O2.
 Growth of these bacteria is not affected by the presence of O2.
 These bacteria have only fermentative type of metabolism.
 Example: Lactobacillus

4. Microaerophiles:

 Those bacteria that do not require O2 for growth but can tolerate low
concentration of O2.
 At atmospheric level of Oxygen growth of these bacteria is inhibited.
 These bacteria only have oxidative type of metabolism
 Example: Campylobacter

5. Obligate anaerobes:
 Those bacteria that can grow only in absence of Oxygen.
 Oxygen is harmful to obligate anaerobes
 These bacteria have only fermentative type of metabolism
 Examples: Peptococcus
Dr. Taghreed Khudhur Mohammed

6. Capnophiles:
 Those bacteria that require carbon dioxide for growth.
 They are CO2 loving organism
 Most of the microaerophilic are capnophilic in nature.
 Example: Campylobacter, Helicobacter pylori, Brucella abortus

F] Classification of bacteria on the basis of Morphology

1. Coccus:

 These bacteria are spherical or oval in shape

 On the basis of arrangement, cocci are further classified as-
 i) Diplococcus: coccus in pair. Ex. Neissseria gonorrhoae, Pneumococcus
 ii) Streptococcus: coccus in chain. Ex.Streptococcus salivarius
 iii) Staphylococcus: coccus in bunch. Ex. Staphylococcus aureus
 iv) Tetrad: coccus in group of four.
 v) Sarcina: coccus in cubical arrangement of cell. Ex. Sporosarcina

2. Bacilli:

 These are rod shaped bacteria

 On the basis of arrangement, bacilli are further classified as-
 i) Coccobacilli: Ex. Brucella
 ii) Streptobacilli: chain of rod shape bacteria: Ex. Bacillus subtilis,
 iii) Comma shaped: Ex. Vibrio cholarae
Dr. Taghreed Khudhur Mohammed

 iv) Chinese letter shaped: Corynebacterium dephtherae

3. Mycoplasma
 They are cell wall lacking bacteria
 Also known as PPLO (Pleuropneumonia like organism)
 Mycoplasma pneumoniae

4. Spirochaetes: They are spiral shaped bacteria

5. Rickettsiae and Chlamydiae; They are obligate intracellular parasites resemble

more closely to viruses than bacteria

6. Actinomycetes
 They have filamentous or branching structure
 They resemble more closely to Fungi than bacteria
 Example: Streptomyces
Dr. Taghreed Khudhur Mohammed

H] Classification of bacteria on the basis of Flagella

1. Monotrichous bacteria:
 Bacteria having single flagella in one end of cell.
 eg. Vibrio cholera, Pseudomonas aerogenosa

2. Lophotrichous bacteria:
 Bacteria having bundle of flagella in one end of cell.
 eg. Pseudomanas fluroscence

3. Amphitrichous bacteria: Bacteria having single or cluster of flagella at both end

of cell. eg. Aquaspirillium

4. Peritrichous bacteria: Bacteria having flagella all over the cell surface. eg. E.coli,
Salmonella, Klebsiella

5. Atrichous bacteria: bacteria without flagella. Ex. Shigella

I] Classification of bacteria on the basis of Spore

1. Spore forming bacteria: Those bacteria that produce spore during unfavorable
condition. These are further divided into two group

i) Endospore forming bacteria: Spore produced within the bacterial cell.

 Bacillus, Clostridium, Sporosarcina etc

ii) Exospore forming bacteria: Spore produced outside the cell

 Methylosinus
Dr. Taghreed Khudhur Mohammed

2. Non sporing bacteria: those bacteria which do not produce spore. Ex. E. coli,
Salmonella