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Chapter 4
SYSTEMATIC
BACTERIOLOGY (A)
Dr Sonal Saxena
Dr Arpita Saxena
GENERAL CONCEPTS
CLASSIFICATION, NOMENCLATURE, AND TAXONOMY OF BACTERIA
GENERAL CONCEPTS OF PATHOGENESIS AND DISEASE SPECTRUM
NOMENCLATURE AND TAXONOMY OF BACTERIA
Enable easy identification of microbes
Bergey’s manual — systematic classification
Carolus linnaeus introduced classification system consisting of domain, kingdom, phylum, class, order, family, genus and species
Subspecies and serotypes are based on antigens
Clone — derived from a single cell; almost all are identical
Colony – represents one clone
Strain – derived from a single source
Kingdom → Division → Class → Order → Family → Tribe → Genus → Species
CLASSIFICATION
1) Phylogenetic
Represented as a branching tree
One characteristic- for division at each branch or level
2) Adansonian classification
Numerical Taxonomy
All characteristics are taken into account
Several characteristics are compared
3) Molecular or genetic classification
Degree of genetic relatedness
Based on the genes, DNA relatedness
G:C ratio (guanine:cytosine): Constant within same species
Used for classifying viruses.
4) Intraspecies classification
Beyond species level
Diagnostic and epidemiological
Classified as:
Biotypes: Biochemical properties
Serotypes: Antigenic features
Phage types: Bacteriophage susceptibility
Colicin types: Production of bacteriocins
4.Systematicbacteriology(A)...............................................................................
MOLECULAR
EPIDEMIOLOGY
Intraspecies typing by newer techniques to the evolution of
molecular epidemiology
Phenotypic – (expressed characteristics) by:
1) Electrophoretic typing
2) Immunoblotting
• Genotypic (direct analysis of genes, chromosomal and
extra-chromosomal DNA) by:
1) Plasmid profile analysis
2) Restriction endonuclease analysis
3) Southern blotting
4) Polymerase chain reaction
5) Nucleotide sequence analysis
TYPE
CULTURES
Representatives of all species
Maintained by International Reference Labs
Made available for study and comparison
ATCC (American Type Culture Collection)
NCTC — National Collection of Type Cultures (Public Health
England)
GENERAL CONCEPTS
OF PATHOGENESIS
AND DISEASE
SPECTRUM
Bacterial
infections can
be
Acute, life-threatening : bacterial meningitis, anthrax
Chronic, indolent : tuberculosis
Mild infection: Streptococcal sore throat
Primary/ secondary infections
Pathogenesis Chain of events that lead to infection
Interaction between source, host and pathogen
PATHOGENESIS
FIG. 4.2 DIAGRAM SHOWING
THE ENTRY, INVASION AND
DAMAGE TO HOST TISSUE BY
PATHOGENIC BACTERIA
PATHOGENESIS
Exposure to the pathogen(bacteria) and route of entry
Bacteria enters through an appropriate route
Encounter host phagocytes  exhibit tissue tropism  multiplication  production of toxin and enzymes  cell destruction 
tissue damage  prevention of host defence  disease manifestation  clinical features
Routes of entry of some disease-causing bacteria
PATHOGENESIS
Incubation period: time between
exposure (entry of pathogen) to
the first manifestation of symptoms
(and clinical signs)
OVERVIEW OF IMPORTANT
BACTERIA ASSOCIATED WITH
HUMAN HEALTH AND DISEASE
GRAM-POSITIVE
COCCI
STAPHYLOCOCCUS,
STREPTOCOCCUS,
PNEUMOCOCCUS,
ENTEROCOCCUS
ANEROBIC GRAM-
POSITIVE COCCI
OVERVIEW OF
IMPORTANT BACTERIA
ASSOCIATED WITH
HUMAN HEALTH AND
DISEASE
Gram-positive cocci: Staphylococcus, Streptococcus, Pneumococcus,
Enterococcus
Gram-positive bacilli: Bacillus, Corynebacterium, Actinomycetes,
Clostridium
Gram-negative cocci: Neisseria, Moraxella
Gram-negative bacilli: Enterobacterales, Vibrios, Non- fermenters
Miscellaneous bacteria: Mycobacteria, Spirochetes, Mycoplasma,
Chlamydia, Rickettsia
GRAM-POSITIVE COCCI
1) Staphylococci
 Spherical, 1 µm diameter
 Arrangement: Grape-like clusters
 Grows in ordinary culture media
 Produce potent toxins, enzymes and
other metabolites
 Develops resistance to antibiotics
Gram-positive cocci in clusters: Staphylococcus aureus
(Source: Department of Microbiology, Pondicherry
Institute of Medical Sciences[PIMS], Puducherry)
Classification
Staphylococci are classified on basis of the coagulase test as coagulase-positive or -negative.
Staphylococcus aureus is coagulase-positive.
Culture characteristics
Staphylococci grow readily on ordinary media
Temperature range of 10–42°C, the optimum being 37°C
pH-7.4–7.6
4.Systematicbacteriology(A)...............................................................................
4.Systematicbacteriology(A)...............................................................................
GRAM-
POSITIVE
COCCI
Common infections by Staphylococcus aureus
 Localised suppurative lesions: Skin and mucosa
 Disseminated: Bloodstream infections, osteomyelitis,
meningitis, pericarditis pneumonia
Coagulase negative Staphylococci (CoNS)
 S.epidermidis, S.lugdenunsis, S.haemolyticus, S.hominis
 Commensals in the body
 S.epidermidis: Most common CoNS infection in humans
- Biofilm formation: important in pathogenesis
- Healthcare-associated infections in
immunocompromised hosts
4.Systematicbacteriology(A)...............................................................................
Coagulase-negative staphylococci (CoNS)
S. epidermidis
S. lugdunensis
S. haemolyticus
S. hominis
Habitat: Commensals in the throat, anterior nares, the skin of the perineum, and the web- spaces
of the hands.
Virulence of CoNS
Biofilm formation is an important factor in the pathogenesis of infections by S. epidermidis.
A biofilm is an extracellular polysaccharide matrix, which protects bacteria from antibacterial
agents and aids colonisation and resistance of infections.
Common
pathogenic
species
S. saprophyticus may be present on healthy human skin and the
periurethral area and can cause urinary tract infection,
particularly in sexually active young women.
S. epidermidis
Most common cause of CoNS infections in humans
Cause disease when the host’s defences are breached or
healthcare-associated infections in immunocompromised hosts
GRAM-
POSITIVE
COCCI
2) Micrococci
Gram-positive cocci in tetrads, pairs or irregular clusters
Rarely cause infection in healthy hosts
3) Streptococci
Morphology: Streptococci are gram-positive
cocci arranged in chains or pairs.
Culture characteristics: Require blood or serum
for growth and are nutritionally more exacting.
Growth is better under 5–10% CO2, optimum
temperature is 37°C (range 22–42°C).
Habitat: Part of the normal flora of human and
animal bodies. Some of them are human
pathogens.
Classification of
Streptococci
1. Based on oxygen requirement, they are classified as :
Facultative anaerobes
Obligate anaerobes
2. Based on hemolysis produced on sheep or horse blood agar,
streptococci are classified as:
Alpha hemolytic :greenish discolouration around colonies (partial
hemolysis). E.g. viridans group of Streptococci, Pneumococci
Beta hemolytic : clear zone of complete hemolysis. E g. Streptococcus
hemolyticus
Non- hemolytic: Fecal Streptococci (Enterococcus)
Classification of
beta hemolyitic
streptococci
Lancefield’s grouping
Based on the carbohydrate antigen (C) on the cell wall,
Rebecca Lancefield (1933) serologically grouped beta (β)-
hemolytic streptococci.
These groups are known as Lancefield’s groups. So far, 20 of
these groups have been identified, named A–H and K–V
(without I and J).
Griffith typing:
It is used for the further classification of S. pyogenes.
Based on the M proteins on the cell surface, they are
subdivided into eighty M types (types M1, M2, M3, and so
on).
Hemolytic
streptococci
Alpha (α)-hemolytic streptococci
They produce a greenish discolouration with partial hemolysis
around the colonies.
The zone of lysis is small( 1or2mmwide).
Most alpha-hemolytic streptococci are commensals in the throat
and may cause opportunistic infections in rare cases.
Examples: Members of the viridans group of streptococci and
pneumococci.
Beta (β)-hemolytic streptococci
They produce a sharply defined, clear, colourless zone of complete
hemolysis, 2–4 mm wide, around their colonies.
The term ‘hemolytic streptococci’ applies strictly to beta-hemolytic
strains.
Most pathogenic streptococci belong to this group.
Infections
caused by
streptococci
STREPTOCOCCUS PYOGENES
& STREPTOCOCCUS
AGALACTIAE ARE IMPORTANT
ONES
4.Systematicbacteriology(A)...............................................................................
Infections
Streptococcus pyogenes or Group A Streptococcus
These are human pathogens, which produce a diverse
array of diseases, ranging from self-limited conditions
(pharyngitis, impetigo, and erysipelas) to severe, invasive
infections (bacteremia, necrotising asciitis, myositis, and
streptococcal toxic shock syndrome.
Streptococcal pharyngitis or skin infections may have
lasting sequelae such as post-streptococcal rheumatic
disease or glomerulonephritis respectively.
Group B streptococci (GBS)—
Streptococcus agalactiae
It is normal inhabitant of the vaginal tract and is associated with neonatal meningitis and
septicemia.
It is also an important pathogen of cattle, producing bovine mastitis.
Nine capsular serotypes have been identified, antibodies to which confer type-specific
protection.
Pneumococci (S. Pneumoniae)
Gram-positive diplococci; capsulated
Flame-shaped/lanceolate appearance
India ink stain: Capsule visualisation
Culture characteristics
 S. pneumoniae have complex growth requirements. They grow
only in enriched media such as sheep blood agar and chocolate
agar with 5–10% CO2.
 Growth appears as 0.5–1 mm, dome-shaped, glistening
colonies showing alpha-hemolysis, which subsequently
become flat with raised edges, central umbonation, and
concentric rings (Draughtsman or carom coin appearance).
Habitat
 S. pneumoniae is found in the throats of approximately half the
population sampled at any time.
Enterococci- electron micrograph showing oval cells
arranged in pairs (Source: Public Health Image Library,
ID266/ Janice Haney Carrr/ CDC)
Infections and
virulence factors
Infections
Meningitis
Lobar pneumonia
Bacteremia
Enterococci
Enterococcus group has E. faecalis, E. faecium, E. durans
Oval diplococci (cocci at an angle to each other), non-capsulated.
Usually non-hemolytic (can be alpha or beta hemolytic)
Habitat
E. faecalis is most often isolated from human sources; it is a part of the
microbiota of the human gut. Identification
They are identified by their ability to grow on bile and cause esculin
hydrolysis, do not grow in 6.5% NaCl, grow on can tellurite blood agar, on
which they produce black colonies.
Infections
Urinary tract infection
Wound infection
Endocarditis
Infections of biliary tract, septicemia and intra- abdominal abscesses
complicating diverticulitis and peritonitis
4.Systematicbacteriology(A)...............................................................................
ANAEROBIC
GRAM-
POSITIVE
COCCI
Peptostreptococci: Normal flora of vagina, intestines and
mouth
Morphology: Relatively small cocci (0.2– 2.5 μm).
Culture: Many of them are aerotolerant and grow well under
10% CO2 in an aerobic atmosphere.
Species: Peptostreptococcus anaerobius P. magnus. P.
asaccharolyticus, P. tetradius, and P. prevotii.
Infections
Puerperal sepsis (Peptostreptococcus anaerobius)
other genital infections
Wound infections, UTI
Osteomyelitis
Abscesses of internal organs (P. magnus)

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4.Systematicbacteriology(A)...............................................................................

  • 1. Chapter 4 SYSTEMATIC BACTERIOLOGY (A) Dr Sonal Saxena Dr Arpita Saxena
  • 2. GENERAL CONCEPTS CLASSIFICATION, NOMENCLATURE, AND TAXONOMY OF BACTERIA GENERAL CONCEPTS OF PATHOGENESIS AND DISEASE SPECTRUM
  • 3. NOMENCLATURE AND TAXONOMY OF BACTERIA Enable easy identification of microbes Bergey’s manual — systematic classification Carolus linnaeus introduced classification system consisting of domain, kingdom, phylum, class, order, family, genus and species Subspecies and serotypes are based on antigens Clone — derived from a single cell; almost all are identical Colony – represents one clone Strain – derived from a single source Kingdom → Division → Class → Order → Family → Tribe → Genus → Species
  • 4. CLASSIFICATION 1) Phylogenetic Represented as a branching tree One characteristic- for division at each branch or level 2) Adansonian classification Numerical Taxonomy All characteristics are taken into account Several characteristics are compared 3) Molecular or genetic classification Degree of genetic relatedness Based on the genes, DNA relatedness G:C ratio (guanine:cytosine): Constant within same species Used for classifying viruses. 4) Intraspecies classification Beyond species level Diagnostic and epidemiological Classified as: Biotypes: Biochemical properties Serotypes: Antigenic features Phage types: Bacteriophage susceptibility Colicin types: Production of bacteriocins
  • 6. MOLECULAR EPIDEMIOLOGY Intraspecies typing by newer techniques to the evolution of molecular epidemiology Phenotypic – (expressed characteristics) by: 1) Electrophoretic typing 2) Immunoblotting • Genotypic (direct analysis of genes, chromosomal and extra-chromosomal DNA) by: 1) Plasmid profile analysis 2) Restriction endonuclease analysis 3) Southern blotting 4) Polymerase chain reaction 5) Nucleotide sequence analysis
  • 7. TYPE CULTURES Representatives of all species Maintained by International Reference Labs Made available for study and comparison ATCC (American Type Culture Collection) NCTC — National Collection of Type Cultures (Public Health England)
  • 8. GENERAL CONCEPTS OF PATHOGENESIS AND DISEASE SPECTRUM Bacterial infections can be Acute, life-threatening : bacterial meningitis, anthrax Chronic, indolent : tuberculosis Mild infection: Streptococcal sore throat Primary/ secondary infections Pathogenesis Chain of events that lead to infection Interaction between source, host and pathogen
  • 9. PATHOGENESIS FIG. 4.2 DIAGRAM SHOWING THE ENTRY, INVASION AND DAMAGE TO HOST TISSUE BY PATHOGENIC BACTERIA
  • 10. PATHOGENESIS Exposure to the pathogen(bacteria) and route of entry Bacteria enters through an appropriate route Encounter host phagocytes  exhibit tissue tropism  multiplication  production of toxin and enzymes  cell destruction  tissue damage  prevention of host defence  disease manifestation  clinical features Routes of entry of some disease-causing bacteria
  • 11. PATHOGENESIS Incubation period: time between exposure (entry of pathogen) to the first manifestation of symptoms (and clinical signs)
  • 12. OVERVIEW OF IMPORTANT BACTERIA ASSOCIATED WITH HUMAN HEALTH AND DISEASE GRAM-POSITIVE COCCI STAPHYLOCOCCUS, STREPTOCOCCUS, PNEUMOCOCCUS, ENTEROCOCCUS ANEROBIC GRAM- POSITIVE COCCI
  • 13. OVERVIEW OF IMPORTANT BACTERIA ASSOCIATED WITH HUMAN HEALTH AND DISEASE Gram-positive cocci: Staphylococcus, Streptococcus, Pneumococcus, Enterococcus Gram-positive bacilli: Bacillus, Corynebacterium, Actinomycetes, Clostridium Gram-negative cocci: Neisseria, Moraxella Gram-negative bacilli: Enterobacterales, Vibrios, Non- fermenters Miscellaneous bacteria: Mycobacteria, Spirochetes, Mycoplasma, Chlamydia, Rickettsia
  • 14. GRAM-POSITIVE COCCI 1) Staphylococci  Spherical, 1 µm diameter  Arrangement: Grape-like clusters  Grows in ordinary culture media  Produce potent toxins, enzymes and other metabolites  Develops resistance to antibiotics Gram-positive cocci in clusters: Staphylococcus aureus (Source: Department of Microbiology, Pondicherry Institute of Medical Sciences[PIMS], Puducherry)
  • 15. Classification Staphylococci are classified on basis of the coagulase test as coagulase-positive or -negative. Staphylococcus aureus is coagulase-positive. Culture characteristics Staphylococci grow readily on ordinary media Temperature range of 10–42°C, the optimum being 37°C pH-7.4–7.6
  • 18. GRAM- POSITIVE COCCI Common infections by Staphylococcus aureus  Localised suppurative lesions: Skin and mucosa  Disseminated: Bloodstream infections, osteomyelitis, meningitis, pericarditis pneumonia Coagulase negative Staphylococci (CoNS)  S.epidermidis, S.lugdenunsis, S.haemolyticus, S.hominis  Commensals in the body  S.epidermidis: Most common CoNS infection in humans - Biofilm formation: important in pathogenesis - Healthcare-associated infections in immunocompromised hosts
  • 20. Coagulase-negative staphylococci (CoNS) S. epidermidis S. lugdunensis S. haemolyticus S. hominis Habitat: Commensals in the throat, anterior nares, the skin of the perineum, and the web- spaces of the hands. Virulence of CoNS Biofilm formation is an important factor in the pathogenesis of infections by S. epidermidis. A biofilm is an extracellular polysaccharide matrix, which protects bacteria from antibacterial agents and aids colonisation and resistance of infections.
  • 21. Common pathogenic species S. saprophyticus may be present on healthy human skin and the periurethral area and can cause urinary tract infection, particularly in sexually active young women. S. epidermidis Most common cause of CoNS infections in humans Cause disease when the host’s defences are breached or healthcare-associated infections in immunocompromised hosts
  • 22. GRAM- POSITIVE COCCI 2) Micrococci Gram-positive cocci in tetrads, pairs or irregular clusters Rarely cause infection in healthy hosts
  • 23. 3) Streptococci Morphology: Streptococci are gram-positive cocci arranged in chains or pairs. Culture characteristics: Require blood or serum for growth and are nutritionally more exacting. Growth is better under 5–10% CO2, optimum temperature is 37°C (range 22–42°C). Habitat: Part of the normal flora of human and animal bodies. Some of them are human pathogens.
  • 24. Classification of Streptococci 1. Based on oxygen requirement, they are classified as : Facultative anaerobes Obligate anaerobes 2. Based on hemolysis produced on sheep or horse blood agar, streptococci are classified as: Alpha hemolytic :greenish discolouration around colonies (partial hemolysis). E.g. viridans group of Streptococci, Pneumococci Beta hemolytic : clear zone of complete hemolysis. E g. Streptococcus hemolyticus Non- hemolytic: Fecal Streptococci (Enterococcus)
  • 25. Classification of beta hemolyitic streptococci Lancefield’s grouping Based on the carbohydrate antigen (C) on the cell wall, Rebecca Lancefield (1933) serologically grouped beta (β)- hemolytic streptococci. These groups are known as Lancefield’s groups. So far, 20 of these groups have been identified, named A–H and K–V (without I and J). Griffith typing: It is used for the further classification of S. pyogenes. Based on the M proteins on the cell surface, they are subdivided into eighty M types (types M1, M2, M3, and so on).
  • 26. Hemolytic streptococci Alpha (α)-hemolytic streptococci They produce a greenish discolouration with partial hemolysis around the colonies. The zone of lysis is small( 1or2mmwide). Most alpha-hemolytic streptococci are commensals in the throat and may cause opportunistic infections in rare cases. Examples: Members of the viridans group of streptococci and pneumococci. Beta (β)-hemolytic streptococci They produce a sharply defined, clear, colourless zone of complete hemolysis, 2–4 mm wide, around their colonies. The term ‘hemolytic streptococci’ applies strictly to beta-hemolytic strains. Most pathogenic streptococci belong to this group.
  • 27. Infections caused by streptococci STREPTOCOCCUS PYOGENES & STREPTOCOCCUS AGALACTIAE ARE IMPORTANT ONES
  • 29. Infections Streptococcus pyogenes or Group A Streptococcus These are human pathogens, which produce a diverse array of diseases, ranging from self-limited conditions (pharyngitis, impetigo, and erysipelas) to severe, invasive infections (bacteremia, necrotising asciitis, myositis, and streptococcal toxic shock syndrome. Streptococcal pharyngitis or skin infections may have lasting sequelae such as post-streptococcal rheumatic disease or glomerulonephritis respectively.
  • 30. Group B streptococci (GBS)— Streptococcus agalactiae It is normal inhabitant of the vaginal tract and is associated with neonatal meningitis and septicemia. It is also an important pathogen of cattle, producing bovine mastitis. Nine capsular serotypes have been identified, antibodies to which confer type-specific protection.
  • 31. Pneumococci (S. Pneumoniae) Gram-positive diplococci; capsulated Flame-shaped/lanceolate appearance India ink stain: Capsule visualisation Culture characteristics  S. pneumoniae have complex growth requirements. They grow only in enriched media such as sheep blood agar and chocolate agar with 5–10% CO2.  Growth appears as 0.5–1 mm, dome-shaped, glistening colonies showing alpha-hemolysis, which subsequently become flat with raised edges, central umbonation, and concentric rings (Draughtsman or carom coin appearance). Habitat  S. pneumoniae is found in the throats of approximately half the population sampled at any time. Enterococci- electron micrograph showing oval cells arranged in pairs (Source: Public Health Image Library, ID266/ Janice Haney Carrr/ CDC)
  • 33. Enterococci Enterococcus group has E. faecalis, E. faecium, E. durans Oval diplococci (cocci at an angle to each other), non-capsulated. Usually non-hemolytic (can be alpha or beta hemolytic) Habitat E. faecalis is most often isolated from human sources; it is a part of the microbiota of the human gut. Identification They are identified by their ability to grow on bile and cause esculin hydrolysis, do not grow in 6.5% NaCl, grow on can tellurite blood agar, on which they produce black colonies. Infections Urinary tract infection Wound infection Endocarditis Infections of biliary tract, septicemia and intra- abdominal abscesses complicating diverticulitis and peritonitis
  • 35. ANAEROBIC GRAM- POSITIVE COCCI Peptostreptococci: Normal flora of vagina, intestines and mouth Morphology: Relatively small cocci (0.2– 2.5 μm). Culture: Many of them are aerotolerant and grow well under 10% CO2 in an aerobic atmosphere. Species: Peptostreptococcus anaerobius P. magnus. P. asaccharolyticus, P. tetradius, and P. prevotii. Infections Puerperal sepsis (Peptostreptococcus anaerobius) other genital infections Wound infections, UTI Osteomyelitis Abscesses of internal organs (P. magnus)