BACTERIOLOGY LAB

As per recommended by the CDC, laboratories should follow the _______ wherein all specimens are treated as potentially infectious

universal precautions

Laboratory personnel are exposed to all potential hazards in the work setting.

Electric Shock

Toxic Gas

Flammable liquids

Radioactive materials

Corrosive substances

Mechanical trauma

Infectious agents

Working area must be cleaned with ++++ dilution of +++++++++ before and after laboratory exercises.

1:10

5% sodium hypochlorite

Any spillage of **infectious agents** must be flooded and cleaned with +++ dilution of

1:5 dilution of 5% sodium hypochlorite

for **infectious** and **contaminated** materials such as used __**cotton, tissue, gloves, culture media,**__ and used disposable __**Petri dishes**__

Yellow plastic bag

for noninfectious dry materials

Black plastic bag

for contaminated sharps like needles

Red puncture proof container

Provide all employees (including the clinical laboratory personnel) with a safe work environment.

Occupational Safety and Health Act (OSHA)

On-site inspection, mandatory standards

Occupational Safety and Health Act (OSHA)

General laboratory safety and infection control guidelines

Clinical and Laboratory Standards Institute (CLSI)

Set min standards for regulation of health facilities and services in the Philippines

Issue permits to construct, License to Operate and Cert. of Accreditation

DOH-HFSRB

aerosols may form during centrifugation of unstopped tubes or from heating cultures or specimens too rapidly (sterilization of inoculating loops in the Bunsen burner flame), removing stoppers from tubes, or leakage from a container that holds contaminated specimens

Airborne

infection may occur as a result of failure to wash hands or eating, drinking, smoking, applying cosmetics, or pipetting with the mouth

Ingestion

infection may result from needlesticks, broken glass, animal bites, or small scratches on the fingers

Direct inoculation

infection may occur if the organism can directly enter through the mucous membranes, such as through the conjunctiva of the eye

Mucous membrane contact

infectious sources include ticks, fleas, and mosquitoes, which may harbor various microorganisms

Arthropod vector

Orientation and continuing education for employees

Safety education

policy and procedures

Safety manual

set of preventive measures designed to reduce the risk of HIV, HBV, and other bloodborne pathogens in the health care setting

universal precautions

apply to all human blood and all other body fluids that contain visible blood

Universal standard precaution

Combination of Univ Precautions & Body Substance Isolation to minimize risk of infection transmission

Standard precautions

All blood, tissue, body fluid, secretions and excretions (except sweat) are considered potentially infectious diseases

Standard precautions

A device that encloses a workspace in such a way as to protect workers from aerosol

Biosafety cabinet

Air that contains the infectious material is sterilized by ++++, +++++ or by passage through a +++++ ++++

Heat

UV light

HEPA filter

allow room (unsterilized) air to pass into the cabinet and around the area and material within, sterilizing only the air to be exhausted

BSC Class I

Common BSC/BSL in the microbiology laboratory

BSC Class IIA, BSL 2

sterilize air that flows over the infectious material, as well as air to be exhausted

BSC Class II

is self-contained, and 70% of the air is recirculated into the work area

Class II Type A

discharged outside the building

Class II Type B

can operate as either a Type A cabinet  when in recirculating mode or a Type B cabinet when exhausting

Class II Type C

When in recirculating mode

Type A cabinet

When exhausting

Type B cabinet

completely enclosed, ventilated, with **negative pressure**, leak-tight construction and attached rubber gloves.

BSC Class III

**In BSC Class III, Supply air** is drawn in through +++++ +++++

HEPA filters

**In BSC Class III, Exhaust air** is treated with either +++++ +++++ +++++ or ++++ +++++ and +++++

double HEPA filtration or HEPA filtration and incineration

No known pathogenic potential for immunocompetent individuals.

BSL I

E ample of pathogen in a BSL 1 Laboratory

Bacillus subtilis

Most **undergraduate laboratory** courses operate under +++++ precautions.

BSL 1

In BSL 1, precautions include adherence to

standard laboratory techniques

process that kills all forms of microbial life, including bacterial endospores.

Sterilization

process that destroys pathogenic organisms, but not necessarily all microorganisms, endospores, or prions

Disinfection

the removal of pathogenic microorganisms so items are safe to handle or dispose

Decontamination

FACTORS AFFECTING THE CONTROL OF MICROBIAL GROWTH

Type of Organism

Microbial Load

Concentration of the Agent to be Used

Presence of Organic Material

Nature of Surface to be Disinfected

Contact Time

Temperature and pH

Compatibility of the Disinfectants

The ability of the microorganisms to withstand the different disinfecting or sterilizing agents depends on the chemical composition and protective mechanisms inherent to such microorganism.

Type of organism

\+++++++ are resistant to many physical and chemical agents because of the lipid-rich mycolic acid present on their cell wall.

Mycobacteria

\+++++++ are produced by some bacteria to confer resistance to heat, radiation, and other harsh environmental conditions.

Endospores

Some bacteria form +++++ that protect them from radiation, desiccation, high pressure, extreme pH extreme temperature, and antimicrobial agents.

biofilms

\++++++++ +++++ refers to the number of microorganisms present in the specimen/object.

microbial load

If the microbial load is high, ++++++++++++ is required to prevent the growth or kill the microorganism.

longer exposure time to the physical or chemical agent

Presence of mucus, blood, and pus can +++++ the disinfecting agent by preventing the full contact of the agent and the material to be disinfected.

inactivate

Examples of chemical agents that can be inactivated include

bleach

QUATS

\++++++ +++++ devised a logical approach for disinfection/sterilization of inanimate objects or surfaces based on the degree of risk involved in their use. This strategy is vital in simplifying how to decontaminate surfaces and maintaining health and safety of the patients.

Earle Spaulding

According to Spaulding classification, equipment/items are regarded as:

critical

semi-critical

non-critical

+++++ ++++ are devices that enter sterile body parts, including the vascular system.

Critical items

\+++++ +++++ items are those that come in contact with non-intact skin or mucous membrane but does not penetrate them.

Semi-critical

\+++++ ++++ items touch only the intact skin but not the mucous membrane.

Non-critical

High level of disinfection is required for the +++++ items, while low level of disinfection is implemented for +++++ items

critical

non-critical

\+++++ +++ refers to the length of time a chemical agent needs to stay on a targeted surface in order to become an effective disinfectant/sterilant. Applying the chemical agent and wiping it immediately may render it useless in killing or inhibiting growth of microorganisms.

Contact time

Most disinfectants are effective at +++++ temperature under an acidic condition.

room

Disinfection can be enhanced by increase in ______ only up to a certain degree. Too much increase in temperature may affect the efficacy of a disinfectant and therefore +++++ its germicidal activity.

temperature

weaken

In chlorination, as pH goes up, the concentration of hypochlorite ions ++++++ and disinfection becomes +++++ effective.

increases

less

\++++ and +++++ should not be used together. Mixing bleach and ammonia may result to a creation of ++++ ++++ that may cause serious injuries.

QUATS

bleach

toxic gas

\++++++ and +++++ are methods for decontamination.

sterilization

disinfection

\++++++ refers to the destruction of all microorganisms, including bacterial spores.

sterilization

\++++++ is the process of destroying pathogenic microorganisms on inanimate objects.

Disinfection

all microbiological wastes must be +++++ prior to disposal

decontaminated

biohazardous wastes shpuld be placed in+++++ bags

2 leak-proof

place contaminated pipettes, swabs, sharps, and glass in ++++++++ containers

puncture-proof resistant

In post-exposure plan, all accidents must be reported to the ++++++ ++++ ++++/+++ ++++

laboratory safety officer

lab supervisor

In fire safety, instruments, appliances must be checked for hazards at least

once every 12 months

ordinary combustible

water

foam

dry chemical

flammable liquids

foam

dry chemical

live electrical equipment

carbon dioxide dry chemical

combustible metals

dry powder

cooking oils

potassium-based alkaline

PASS

Pull Pin

Aim Nozzle

Squeeze Trigger

Sweep Nozzle

max limits that a worker can continuously be exposed to a chemical up to +++++ mins without health danger

short-term exposure limits

15 mins

Manufacturer's name and \n address, emergency telephone \n number, telephone number for \n information, date prepared

Sec 1

Hazardous ingredient's identity \n information, components, \n specific chemical identity \n common name(s), PEL

Sec 2

Physical/chemical \n characteristics: boiling point, \n specific gravity, vapor \n pressure, vapor density, \n melting point, evaporation rate, \n solubility in water, appearance \n and odor

Sec 3

Fire and explosion hazard \n data: flash point, flammable \n limits extinguishing media, \n special firefighting procedures, \n unusual fire and explosion \n hazards

Sec 4

Reactivity data: stability \n incompatibility, hazardous \n decomposition by-products, \n hazardous polymerization

Sec 5

Health hazard data: route(s) of \n entry inhalation, skin, \n ingestion, health hazards \n (acute and chronic), \n carcinogenicity, signs and \n symptoms of exposure, \n medical conditions, generally \n aggravated by exposure, \n emergency and first-aid \n procedures

Sec 6

Precautions for safe handling \n and use: waste-disposal \n method, precautions to be \n taken in handling and storing, \n action if material is released or \n spilled

Sec 7

Control measures: respiratory \n protection, ventilation, eye \n protection, gloves, \n work/hygienic practices

\n

Sec 8

Collect specimen in the \n

right time, right place

Collect specimen in the +++++ phase of the infection

acute

Collect specimen prior to administration of ++++

antibiotics

Use Sterile Containers except in ++++ +++++

Stool Collection

In collecting lesions, wounds, abscess, collect at the

margin (swabs)

sueprficial wound

swab

deep wound

aspirate

swab is for the recovery of +++++ organisms

aerobic

two swabs are used: one is for ++++ and one is for ++++

gram staining

inoculation of culture

swab is made out of ++++

cotton

swabs for bacterial culture

Dacron

Calcium Alginate

Rayon

swabs for viral culture

Cotton

Dacron

swabs for nasopharyngeal seab

flexible swab

shaft swab

plastic or wooden

throat cultures are for the isolation of

Group A Streptococcus