Simple Guide Identification Bacteria: Dr. Khalil Alkuwaity
Simple Guide Identification Bacteria: Dr. Khalil Alkuwaity
Simple Guide Identification Bacteria: Dr. Khalil Alkuwaity
A simple guide for the
identification of bacteria
Dr. Khalil Alkuwaity
CONTENTS
Introduction 2
Abbreviations & shapes 3
The Family Stone 4
The Big Family 5
Gram Positive Cocci 6
Gram Negative Bacilli 8
Gram Positive Bacilli 9
Gram Negative Cocci 10
Gram Negative Coccobacilli 11
Anaerobic Bacteria 12
Acid Fast Bacilli 13
The Big Gathering 14
Member Profile 15
The Normal Flora 16
The Usual Suspects 17
Prepared by
DR. KHALIL ALKUWAITY
Department of Medical Laboratory Technology
Faculty of Applied Medical Sciences ‐ KAU
(December, 2012).
DR. ALKUWAITY 1 ONE BIG FAMILY
INTRODUCTION
The purpose of this booklet is to provide a quick and easy
guide to the classification of clinically important bacteria.
The classification is principally based on the GRAM STAIN
result of the organism. Further diagnostic tests include:
BIOCHEMICAL reactions AND sensitivity to ANTIBIOTICS.
***
REMEMBER that good IDENTIFICATION always begins with
SEPARATED COLONIES and a clear GRAM STAIN result.
1 CULTURE
Gram‐negative
Gram‐negative coccobacilli
cocci in pairs
Gram‐positive
cocci in clusters
Gram‐positive
cocci in pairs
2 STAIN
Gram‐positive
cocci in chains
Gram‐negative
bacilli
Gram‐positive Yeast cell
bacilli (budding)
DR. ALKUWAITY 2 ONE BIG FAMILY
ABBREVIATIONS & SHAPES
The following ABBREVIATIONS and SHAPES are used throughout this booklet:
GPB Gram Positive Bacilli
GPC Gram Positive Cocci
GNB Gram Negative Bacilli
GNC Gram Negative Cocci
GNCB Gram Negative Coccobacilli
AFB Acid Fast Bacilli
DNase positive
Bile Esculin positive
Lactose fermenter
Non‐lactose fermenter
Low virulence organism
S Sensitive
R Resistant
O2 Strict aerobic (O2 present)
CO2 Capnophilic (5% CO2)
MO2 Microaerophilic (Low O2)
Ø2 Strict anaerobic (O2 absent)
Spore‐forming
Motile
Diplococci
Likely to produce β‐lactamases
Curved bacilli
DR. ALKUWAITY 3 ONE BIG FAMILY
THE FAMILY STONE
The following diagrams compare the CELL WALL structure of
Gram‐positive, Gram‐negative and Acid‐fast bacteria:
GRAM‐POSITIVE CELL WALL
Peptidoglycan (>60%)
Cytoplasmic membrane
GRAM‐NEGATIVE CELL WALL
Outer membrane → Lipopolysaccharide
Periplasmic space
Peptidoglycan (5‐10%)
Periplasmic space
Cytoplasmic membrane
ACID‐FAST CELL WALL
Glycolipids → Mycolic acid (60%)
Arabinogalactan
Peptidoglycan
Cytoplasmic membrane
Cell wall
Cell membrane
Cytoplasm
DR. ALKUWAITY 4 ONE BIG FAMILY
THE BIG FAMILY
AFB
GPB GPC
Bacteria
GNB GNC
GNCB
NOTES
When stained using the Gram stain: GRAM‐POSITIVE bacteria appear PURPLE, while
GRAM‐NEGATIVE bacteria appear PINK.
ACID FAST BACILLI (AFB) are stained using special dyes such as the Ziehl‐Neelson
stain and they appear as RED bacilli against a blue background.
In addition, there are other types of bacteria that belong to different groups such as
the Spirochetes, Chlamydia and Mycoplasmas. These are not included in this guide,
as their diagnosis is based on different detection or culture techniques.
DR. ALKUWAITY 5 ONE BIG FAMILY
Gram Positive Cocci
GPC
_
Catalase
+
Micrococcaceae
O2
Micrococcus
Staphylococci
Coagulase
_
+
S. aureus CoNS*
Methicillin Novobiocin
S R
R
S. epidermidis
MRSA
S. saprophyticus
Methicillin Resistant
Staphylococcus Aureus This distinction is usually made in
urinary tract infections.
*
Coagulase Negative Staphylococci
DR. ALKUWAITY 6 ONE BIG FAMILY
Gram Positive Cocci
...continued
Catalase ̶
Streptococcaceae
6.5% NaCl +
Enterococci
Vancomycin
R
VRE
Streptococci
Colony on Blood Agar
Beta hemolysis Alpha hemolysis
OR
Bacitracin: S
Group A
Lancefield
Optochin
Grouping
CAMP +
Group B
S
S. pneumoniae
6.5% NaCl ̶
Group D
R
Viridans Streptococci
Group A = Streptococcus pyogenes Enterococci include E. faecalis, E. faecium
Group B = Streptococcus agalactiae VRE: Vancomycin Resistant Enterococci
Group D = Non‐Enterococci Viridans Strept include S. mitis, S. mutans
DR. ALKUWAITY 7 ONE BIG FAMILY
Gram Negative Bacilli
Swarming, Urease +
Proteus
spp.
Indole + XLD → Black center
Escherichia coli Salmonella spp.
ESBL**
Klebsiella spp. Shigella spp.
Enterobacteriaceae*
_
API 20E OR
Oxidase GNB Analytical VITEK
Profile Index
+
O2 Pyocyanin TCBS media
Pseudomonas aeruginosa Vibrio cholerae
MO2 MO2
Helicobacter pylori Campylobacter jejuni
Urease +
*
Other important
**
May produce ESBL enzymes Enterobacteriaceae include:
(Extended Spectrum Beta‐Lactamases) Citrobacter freundii
=> Resistance to cephalosporins. Enterobacter cloacae
Morganella morganii
Serratia marcescens
Yersinia enterocolitica
Identify using API or VITEK
DR. ALKUWAITY 8 ONE BIG FAMILY
Gram Positive Bacilli
B. cereus
Psychrophile
L. monocytogenes B. anthracis
Listeria Bacillus
All
Lactobacilli GPB Catalse +
Catalase
Corynebacterium Diphtheroids
C. diphtheriae
Chinese letters
Tellurite‐blood agar
(Toxigenic) Elek test +
DR. ALKUWAITY 9 ONE BIG FAMILY
Gram Negative Cocci
GNC
CO2 All
Neisseria spp Neisseriaceae Oxidase +
Neisseria gonorrhoeae Neisseria meningitidis
Glucose fermenter Glucose/maltose fermenter
Thayer Martin agar
Moraxella catarrhalis
Non‐fermenter
DR. ALKUWAITY 10 ONE BIG FAMILY
Gram Negative Coccobacilli
XV
H. influenzae
V X
H. parainfluenzae H. ducreyi
V&X
Factors
CO2
Haemophilus
All
GNCB Catalase +
Oxidase +
Brucella Bordetella
Serology O2
B. pertussis
B. melitensis
Charcoal‐blood agar
B. abortus co2
B. suis
DR. ALKUWAITY 11 ONE BIG FAMILY
Anaerobic Bacteria
C. tetani C. botulinum
C. perfringens C. difficile
Naglar reaction Toxin-A test
Clostridium
GPB
Ø
2
GNB
Bacteroides
B. fragilis
Metronidazole: S
DR. ALKUWAITY 12 ONE BIG FAMILY
Acid Fast Bacilli
AFB
AFB
Mycobacterium*
O2
M. tuberculosis
TB rods stained using Ziehl‐Neelsen (ZN) stain and
Auramine‐Rhodamine (AR) fluorescent stain
TB colonies grown on
Lowenstein Jensen (LJ) medium
*
Mycobacterium species include:
Mycobacterium tuberculosis (MTB) => TB infection
Mycobacteria Other Than Tuberculosis (MOTT) => TB‐like infection
DR. ALKUWAITY 13 ONE BIG FAMILY
THE BIG GATHERING
(Complete with the correct names of organisms)
+
B C
DR. ALKUWAITY 14 ONE BIG FAMILY
MEMBER PROFILE
(Growth and colony appearance)
* Shows characteristic morphology, but the organism may also grow on other media.
ABBREVIATIONS:
BAP = Blood Agar Plate, CAP = Chocolate Agar Plate,
MSA = Mannitol Salt Agar, DCA = Deoxycholate Citrate Agar,
MAC = MacConkey agar, XLD = Xylose Lysine Desoxycholate,
LJ = Lowenstein Jenson, TCBS = Thiosulfate Citrate Bile salts Sucrose.
DR. ALKUWAITY 15 ONE BIG FAMILY
THE NORMAL FLORA
(Normally present in healthy individuals)
BODY SITE NORMAL FLORA
Skin, Eye CoNS*
Viridans Streptococci
& Urethra
Diphtheroids
CoNS*
Viridans Streptococci
Oropharynx
Diphtheroids
Neisseria spp
GBS
CoNS*
Vagina
Lactobacilli
Candida spp
Bacteroides
Clostridium
Large intestine Coliforms**
Proteus
Pseudomonas
CSF
BODY FLUIDS
Blood
Urine (in bladder) Normally Sterile
Synovial Fluid
Ascitic Fluid
*
CoNS: Coagulase Negative Staphylococci (e.g. S. epidermidis).
**
Coliforms: are lactose‐fermenting Enterobacteriaceae, such
as E. coli, Klebsiella, Citrobacter and Enterobacter.
DR. ALKUWAITY 16 ONE BIG FAMILY
THE USUAL SUSPECTS
(Common causative agents)
Meningitis
N. meningitidis
Eye infection S. pneumoniae
H. influenzae Pharyngitis
H. influenzae
E. coli Strept Group A,C,G
S. pneumoniae
P. aeruginosa
GBS H. influenzae (<6yrs)
Pneumonia
Septicemia S. pneumoniae
Staphylococci H. influenzae
Streptococci M. catarrhalis
GNB GNB
C. albicans
Anaerobes Skin & Wound
S. aureus
Gastroenteritis S. pyogenes
Salmonella GNB
Shigella Anaerobes
Campylobacter
Helicobacter
Urinary Tract
Vibrio
Infection
E. coli
Genital infection Other Body Fluids Klebsiella
N. gonorrhoeae Staphylococci Proteus
T. pallidum Streptococci Enterococcus
H. ducreyi GNB P. aeruginosa
G. vaginalis Anaerobes S. saprophyticus
C. albicans C. albicans
DR. ALKUWAITY 17 ONE BIG FAMILY