Medical Technology

Assessment Program I
BACTERIOLOGY

[TRANS]: BACTERIAL MORPHOLOGY AND CYTOLOGY

Coccus
OUTLINE
• Chain – Streptococcus
I. Bacterial Morphology & Cytology
• Cluster – Staphylococcus (grape-like cluster)
A. Shape of Bacteria
i. Coccus • One plane of division – diplococci/ cocci in pairs (coffee-
ii. Bacillus bean/ kidney-shape) → will produce Neisseria spp.
iii. Coccobacillus except Neisseria ilongata.
iv. Vibrio o Neisseria spp. – intracellular Gram negative
v. Spiral diplococci.
II. Different Parts of The Cell • One plane of division each coccus continually dividing
A. Bacterial Cell Membrane
forming chains will produce Streptococcus
B. Bacterial Cell Wall
III. Bacterial Structures • Two planes of division forming tetrads or pockets of four
A. Parts Internal to Cell Wall will produce Micrococcus
B. Parts External to Cell Wall • Three planes of division in a regular pattern forming cube-
IV. Pathogenic Determinants like arrangement will produce Sarcinae
• Three or more planes of division in an irregular pattern
BACTERIAL MORPHOLOGY AND CYTOLOGY forming cluster will produce Staphylococcus
Table No. 1 Prokaryotes vs. Eukaryotes Bacillus
PROKARYOTES EUKARYOTES • Elongated
Nuclear Body • Nucleoid – not • Nucleus – bound • Chain – Streptobacillus
bound by a by a membrane. • Short bacilli in a school of fish arrangement –
membrane. • Consists of one Haemophilus ducreyi (smallest pathogenic bacteria).
• Consists of or more paired
single circular chromosomes
• Large and sporulated in a bamboo fishing pole or bamboo
chromosome which consist of fishing rod – Bacillus anthracis (largest pathogenic
which consists DNA and bacteria).
of DNA and histone like • Club-shaped when granules are concentrated on one
histone like CHON. pole. Chinese character in arrangement – C. diphtheriae
CHON. • Comma-shaped bacilli – Vibrio cholerae
Cell division • Binary fission • Mitosis • S-haped, seagull’s wing – Campylobacter spp. and
Cell wall • Eubacteria – • Animals/ Helicobacter spp.
with cell wall Protozoans –
(consists of lack cell wall Coccobacillus
peptidoglycan • Plant/fungi/alga • Short bacillus
except e – with cell wall Vibrio
Mycoplasma • Consists of
and cellulose and • Curved
Ureaplasma) chitin. Spiral
Cytoplasmic • Fluid • Fluid • Spirilla
membrane phospholipid phospholipid
o Spirillum minor – causative agent of rat bite fever.
bilayer without bilayer with CHO
CHO and sterol. and sterol. • Spirochete
Cellular • Absent • Present o Borrelia – flexibly twisted organism resembling
organelles stretched spiral.
Site of energy • Cytoplasmic • Mitochondria
o Leptospira – tightly twisted organisms with one or
production membrane both ends bent into a hook.
Site of CHON • Free ribosomes • Rough ER
o Treponema – tightly twisted resembling corkscrew;
synthesis causative agent of syphilis.
• Note: Leptospira and Treponema are common in the
SHAPE OF BACTERIA Philippines.

DIFFERENT PARTS OF THE CELL

BACTERIAL CELL MEMBRANE
• Bilayer phospholipid
o Hydrophilic region – the layer that attracts water.
o Hydrophobic region – the layer that repels water.
• Water can penetrate
• Flexible
• Not strong, ruptures easily
o Osmotic pressure created by cytoplasm.
o The pressure produced by the water presents in the
Figure No. 1 Shape of Bateria cytoplasm destroys cell membrane.

ALISON, ARCHIVIDO, MASCARINAS, REYES, VICENTE 1

 TRANS: BACTERIAL MORPHOLOGY AND CYTOLOGY

c. Surface O antigen/ Somatic antigen

o V – cystal Violet (primary stain)
o I – Gram’s iodine (mordant)
▪ Gram’s Iodine will form complex with crystal
violet.
o A – cetone-alcohol/ 95% ETOH (decolorizer)
o S – afranin (counter stain)
• Decolorization – most important and critical step in
Gram’s staining.
o Gram Positive Bacteria – has MgRNA + Crystal
Violet + Iodine.
▪ In Gram Positive, crystal violet and Gram’s
Figure No. 2 Bacterial Cell Membrane
iodine form complex with MgRNA. This complex
is insoluble to alcohol, reason why after
BACTERIAL CELL WALL decolorization it retained the violet color.
o Gram Negative Bacteria – it has lipid which is
soluble in alcohol, thus, after decolorization, the
primary stain will be removed.
• Gram staining is performed in order to group bacteria into
Gram Positive and Gram Negative according to their cell
wall and will be further group into cocci or bacilli.

General Rules
Figure No. 3 Gram Positive vs Gram Negative Cell Wall 1. All cocci are Gram Positive EXCEPT:
o Neisseria
• It is composed of 70% water. o Branhamella
• It is composed of 30% carbohydrates, proteins, lipids, and o Veillonella
enzymes. 2. All bacilli are Gram Negative EXCEPT:
• It gives shape to the bacteria. o Mycobacterium
• Some components present in the bacterial cell wall are ▪ Mycobacterium is stained with Acid Fat Staining
responsible of for the pathogenicity of the organism. and not with Gram stain. The Gram reaction of
o Pathogenic determinants – structures that will give Mycobacterium is Gram Positive.
the microorganisms additional protection, thus, o Bacillus
having high pathogenicity. o Corynebacterium
o Examples: o Erysipelothrix
▪ M Protein – responsible for the resistance of the o Listeria
organism to phagocytosis (Streptococcus o Lactobacillus
pyogenes). o Rothia
o The cell wall of Streptococcus pyogenes is o Kurthia
embedded with M protein which help the 3. Spiral organisms are very hard to stain. Stainable spiral
organism to evade phagocytosis. are Gram Negative.
▪ Mycolic Acid – responsible for the resistance of o Spirochetes such as Treponema and Leptospira are
organism to digestion during phagocytosis stained using fluorescent dyes.
(Mycobacterium); present in the cell wall of o Some spirochetes are stainable and their Gram
Mycobacterium spp. reaction is Gram Negative.
o Mycobacterium may be phagocytosed but it 4. Mycoplasma and Ureaplasma are Gram Negative,
will resist the digestion process because of although they are cell wall less.
the mycolic acid present in its cell wall.
For Gram Stain Reactions BACTERIAL STRUCTURES
1. Gram-positive (+)
o Have thick peptidoglycan layer (murein,
mucopeptide layer).
o Consists of N–acetylglucosamine and N-acetyl
muramic acid
o Teichoic acid – binds Mg ions.
o The color of the Gram positive bacteria is the color of
primary stain (violet).
2. Gram-negative (–)
o Periplasmic space – between cytoplasmic
membrane and cell wall.
o Gel-like protein
o Thin peptidoglycan layer Figure No. 4 Bacterial Structures
o Thick outer membrane
o The color of the Gram negative bacteria is the color • Flagella
of secondary/ counter stain (red). • Pili
o Produces endotoxin because of the Lipid A present • Capsule
in its outer membrane. • Plasma Membrane
1. Phospholipid layer – innermost • Cytoplasm
2. LPP
• Cell wall
3. Lipopolysaccharide (LPS)
• Lipopolysaccharides
a. Lipid/ Endotoxin – innermost
b. Core polysaccharide layer • Teichoic Acids
• Inclusions

ALISON, ARCHIVIDO, MASCARINAS, REYES, VICENTE 2

 TRANS: BACTERIAL MORPHOLOGY AND CYTOLOGY

• Spores Arrangement Basis for Classification

• Pathogenic determinants include spores, capsule, pili, 1. Atrichous – no flagella
flagella, teichoic acids. 2. Monotrichous – flagellum on one pole
3. Ampitrichous – presence of flagella on both pole
PARTS INTERNAL TO CELL WALL 4. Lopotrichous – with tough or group of flagella on one or
1. Cytoplasmic membrane – selective permeable both poles
membrane, site of energy production 5. Peritrichous – bacterial cell is completely surrounded by
2. Mesosomes – extension of the cytoplasmic membrane, flagella
point of attachment of chromosome 6. Periplasmic flagella/ Axial filament – flagella enclosed
3. Ribosome – tiny particle consist of protein and RNA, site on the periplasmic space (ex. Spirochetes)
of protein synthesis Test for Motility
4. Inclusion bodies – act as storage for nutrients
o Microorganism may survive with or without this 1. Hanging drop
a. Much granules – for Mycobacterium, consist of 2. Semi-solid motility medium (SIM)
lipids 3. Special stains for flagella
b. Metachromatic granules/ volutin granules/ a. Leifson
babes-ernst granules – for Corynebacterium, b. Gray
Diptheriae (consist of polyphosphate) c. Fisher and Conn
c. Bipolar bodies – for Yersinia pestis ▪ All contain TANNIC ACID that will coat, swell
5. Endospores/ bacterial spores and forms a precipitate with flagella.
o Resistant structure
▪ Heat, irradiation, cold PATHOGENIC DETERMINANTS
▪ Boiling > 1 hr still available • Presence of capsule
o Takes time and energy to make spores • Some components of bacterial Cell Wall (Ex. M protein)
o Location important in classification • Enzymes
▪ Central, subterminal, terminal o Coagulase – converts fibrinogen to fibrin clot (coats
o Bacillus stearothermophilus – spores the WBC particularly neutrophil and macrophages
▪ Used for quality control of heat sterilization thus inhibit phagocytosis)
equipment o Kinase/Fibrinolysin – dissolve fibrin clot and once it
▪ Spores can be found on the lines of an autoclave is dissolved, it is responsible for the spread of
tape. Blackening indicates maximum infection
sterilization process (121’C, 15 psi) o Hyaluronidase – hydrolysis of hyaluronic acid in
o Bacillus anthracis spores connective tissue
▪ Used in biological warfare • Bacterial toxins
▪ Can be intestinal, respiratory, or skin
o Consist of calcium dipicolinate/ dipicolinic acid which Table No. 2 Exotoxin vs. Endotoxin
is responsible for the heat resistance of the spore EXOTOXIN ENDOTOXIN
o Spore formers Bacterial source G (+) and G (-) Only G (-) except
▪ Bacillus – sporulate aerobically Mycoplasma and
▪ Clostridium – sporulate anaerobically Ureaplasma
Mechanism Metabolic product Release only
release in the upon lysis of
PARTS EXTERNAL TO CELL WALL surrounding bacteria
1. Glycocalyx environment
o Capsule – organized material firmly attached to cell Composition CHONs, Peptides Lipid A
wall Heat stability Heat labile, destroyed Heat stable,
▪ Function: inhibit phagocytosis by heating at 60-80ºC withstand the
▪ Antigen: K (but for Salmonella typhi: Vi antigen) autoclave temp.
▪ Composition: polysaccharide Immunologic Could be converted to Could not be
o Slime layer – unorganized, not attached to cell wall property toxoid, easily converted to
neutralize with toxoid
2. Pili/ Fimbriae
antitoxin
o Hair-like projections Pharmacologic Specific: Non-specific,
o Composition: protein known as PILIN property generalized
▪ Sex/ fertility pili/ F pili – for sexual conjugation • Cytotoxin – if it
damages host (fever and
▪ Somatic/ common/ ordinary pili – for adhesion of cells shock)
bacterial cell wall • Enterotoxin –
o Note: Bacteria multiplies asexually through binary damages the cell
fission but still capable of sexual conjugation lining the GIT
(exchange of genetic information) through Pili • Neurotoxin –
3. Flagella interfere with
o Slender, whip-like structure nerve impulse
o Function: locomotion transmission
▪ Flagellated microorganisms are more
pathogenic because it can travel from its habitat REFERENCES
to other parts of the body such as Escherichia
coli. Notes from the discussion by Mr. Edison D. Ramos, RMT, MSMT,
MPh
o Composition: protein known as FLAGELLIN
o Antigen: H National University powerpoint presentation: Bacterial Morphology
▪ Somatic antigen – O and Cytology
▪ Capsular antigen – K
o Swarming occurs with some bacteria
▪ Spread across Petri Dish
▪ Proteus species most evident

ALISON, ARCHIVIDO, MASCARINAS, REYES, VICENTE 3