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Microbiology

•• Completely resistant to antibiotics acting on Culture

cell wall (β-lactams)
•• In solid medium (containing PPLO agar) very
•• Pleomorphic: exist in coccoid, bacillary or small (200–500 μm size) Fried egg appearance
Q
filamentous helical forms colonies
•• Poorly gram-negative, better stained by Giemsa
stain Pulmonary Manifestations
•• Reproduce by binary fission and budding Atypical community acquired interstitial pneumonia
Also known as 
•• Eaton agent pneumonia
•• Primary atypical pneumonia
•• Walking pneumonia

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Microbiology
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Microbiology
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peptidase domain of angiotensin converting ARTHROPOD- AND RODENT BORNE

enzyme 2
VIRAL
•• The cellular Protease TMPRSS2 also appears
Chikungunya
important for SARS-COV-2 cell entry.
Dengue
Mosquito-borne Yellow fever
JE
Zika virus
KFD
Tick-borne Crimean Congo hemorrhagic fever
Colorado tick fever
Hanta virus
Lassa fever
Rodent-borne
Hemorrhagic fever with renal
syndrome

PARASITOLOGY - MASTER CHARTS

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Microbiology
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Microbiology

1. Direct Microscopy
•• KOH (10%) mount or histopathological staining
characteristic narrow septate hyaline hyphae
Q
with acute angle branching

Aspergillus
Macroscopic appearance of colony Microscopic appearance of colony (LPCB mount)
species
•• Vesicle is conical-shaped
Colonies—smoky green, velvety to powdery, reverse •• Phialides are arranged in single row
A. fumigatus
is white •• Conidia arise from upper third of vesicle
•• Conidia are hyaline

•• Vesicle is globular-shaped
•• Phialides in one or two rows
A. flavus Colonies—yellow green, velvety, reverse is white
•• Conidia arise from upper two-third to entire vesicle
•• Conidia are hyaline

•• Vesicle is globular-shaped
•• Phialides in two rows
A. niger Colonies—black, cottony type, reverse is white
•• Conidia arise from entire vesicle
•• Conidia are black in color

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Microbiology
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STERILISATION AND
DISINFECTION

Sterilization
•• An article is made free of all microorganisms
either in the vegetative or spore form
•• Complete absence of microorganisms including
spores.

Disinfection 2. Flaming
•• Destruction of all pathogens or organisms •• Objects are passed through bunsen flame
capable of producing infections but not without allowing them to become red hot
necessarily spores
Eg. Glass slides, scalpels and mouths of culture
•• All organisms may not be killed but the number tubes
is reduced to a level that is no longer harmful
to health.
•• Unlike sterilization, disinfection is not sporicidal
(does not kill spores)

Methods of Sterilisation
Physical methods Chemical methods
1. Sunlight 1. Alcohols
2. Heat 2. Aldehydes
a. Dry heat 3. Phenols
b. Moist heat 4. Halogens
3. Ozone 5. Oxidizing agents 3.Incineration
4. Filtration 6. Salts •• Infective material is reduced to ashes by
5. Radiation 7. Surface active agents burning.
8. Dyes •• Eg. Soiled dressings, animal carcasses, bedding
9 Vapor phase Disinfectants and pathological materials are dealt with this
method.

1.Red heat 4. Hot Air Oven

•• Objects are held directly in the flame of a •• Most widely used method of sterilization by dry
Bunsen burner till they become red hot. heat
•• Eg. Inoculating wires, loops, tips of forceps and •• Electrically heated → hot air is created
needles
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Microbiology

•• Fitted with a fan to ensure adequate and even •• Eg. serum or egg media, such as Lowenstein-
Q
distribution of hot air in the chamber Jensen’s and Loeffler’s serum
•• Fitted with a thermostat that maintains the
chamber air at a chosen temperature Tyndallization / Intermittent
sterilization
•• Steam at 100°C for 20 minutes on three
successive days
•• Principle → first exposure kills all the vegetative
forms
•• → In the intervals between the heatings the
remaining spores germinate into vegetative
forms which are killed on subsequent heating.
•• Eg . sterilization of egg, serum or sugar
containing media which are damaged at higher
•• 160°C for two hours (holding time)
Q temperatures of autoclave.

•• 170°C for one hour Steam sterilizer

•• 180°C for 30 minutes •• Steam sterilizer at 100°C for 90 minutes
Uses: •• Articles are kept on a perforated tray through
•• Glasswares like glass syringes, petri dishes, which steam can pass.
Q
flasks, pipettes and test tubes. •• Most of the vegetative forms are killed by this
•• Surgical instruments like scalpels, scissors, method except thermophiles
forceps etc. •• Eg. Media which are decomposed at high
•• Chemicals → oily fluids such as liquid paraffin, temperature of autoclave
fats, grease, dusting powder etc.

Sterilization control
1. Biological control → Spores of “Bacillus subtilis
Q
subsp. Niger ”
2. Chemical indicator → Browne’s tubes No. 3
containing red solution is inserted in each load
and a color change from red to green is observed,
Q
which indicates proper sterilization

Pasteurisation
•• Method for sterilization of milk
•• Kills nearly 90% of the bacteria in milk
•• But it will not kill coxiella burnetii and
bacterial spores.

Two methods are commonly used:

A. Holder method: 63°c for 30 minutes.
Autoclave
Sterilization by steam under pressure (modified
B. Flash method: 72°c for 15-20 seconds Pressure cooker)
Q

•• Temperature — 121 °C
Inspissation
•• Chamber pressure — 15 lb per square inch
•• Heating at 80-85°C temperature for half an
hour daily on three consecutive days •• Holding time —15 minutes
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Principle Ionizing radiations

•• Gamma radiations from a Cobalt 60 source
•• High penetrating power
•• Damage DNA by various mechanisms.
•• Known as cold sterilization because there is no
appreciable increase in temperature.
Eg sterilization of disposable items such as plastic
syringes, swab culture plates, cannulas, catheters
etc
•• Sterilization control: Bacillus pumilis

Uses:
•• Culture media
•• Rubber material
•• Aprons , gowns, dressing, gloves etc.
Biomedical waste management
•• Surgical instruments except sharps.
•• Biomedical Waste Management Rule, India,
•• For all glass syringes, a hot air oven is a 2016 (including the amendment added in 2018
better sterilizing method. and 2019)
–– Yellow
Sterilization controlQ
1. Biological control → Spores of Bacillus –– Red
stearothermophilus –– White
2. Chemical indicators → Browne’s tube contains
–– Blue
red solution which turns green, when exposed to
temperature of 121°C for 15 minutes in autoclave
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Microbiology

Category Type of waste Treatment/ disposal options

Yellow Human anatomical waste Incineration/ Plasma pyrolysis/ deep burial
–– Infectious
–– Non- plastic
waste

Animal anatomical waste

Q
Soiled waste Incineration/plasma pyrolysis/ deep burial/
autoclaving or hydroclaving + shredding/mutilation
Expired/discarded medicines — pharmaceutical Sent back to manufacturer / CBMWTF for
Q
waste, cytotoxic drugs incineration (cytotoxic drugs at temperature
>1200°C)
Chemical solid waste Incineration or plasma pyrolysis or encapsulation
Chemical liquid waste such as discarded disinfectants, Pre-treated before mixing with
infected body fluids and secretions, liquid from house- other wastewater
keeping related activities
–– Discarded linen waste –– Non-chlorinated chemical
–– Contaminated with blood/body fluids, mask, –– Disinfection followed by incineration/plasma
cap, gown and shoe cover pyrolysis
Microbiology, other clinical laboratory waste, blood Pre-treat to sterilize with non
bags, live/ attenuated vaccines chlorinated chemicals2 on-site as per NACO / WHO
guidelines (Blue book 2014) + incineration
Q
Red –– Infectious plastic waste –– Autoclaving/microwaving/hydroclaving +
Non-chlorinated plastic –– Disposable items such as tubing, bottles, intravenous shredding
bags or containers tubes and sets, catheters, urine bags, syringes –– Mutilation/sterilization + shredding
(without needles and fixed needle syringes) –– Treated waste sent to authorized recyclers
and vacutainer with their needles cut), gloves, or for energy recovery
Q
plastic apron and goggles

White (Translucent) Waste sharps including metal sharps Autoclaving/dry heat sterilization followed by:
Puncture-proof, leak- Needles, syringes with fixed needles, needles from • Shredding or mutilation or encapsulation in metal
proof, tamper-proof needle tip cutter or container or cement concrete or
containers burner, scalpels, blades, or any other contaminated • Sanitary landfill or
sharp (used or discarded) • Designated concrete waste sharp pit

Q
Blue Glasswares : Broken or discarded and contaminated Disinfection can be carried out by:
Puncture-proof, leak- glass including medicine vials and ampoules except –– Soaking the washed glass waste after
proof containers those contaminated with cytotoxic wastes, cleaning with detergent and sodium
microscope slides
hypochlorite treatment (1–2%) or
Q
–– Metallic body implants –– Autoclaving/microwaving/ hydroclaving
–– Dental implants, other body implants and and then it is sent for recycling
plates