SAFETY AND QC FOR BACTERIOLOGY

Pathogen

• organism

Reservoir

• people, animals, soil, food, water

Portal of Exit

• coughing, sneezing, body secretion, feces

Mode of Transmission

• direct contact, indirect contact, vectors

Portal of Entry

• mouth, nose, eyes, cuts in skin

True

T or F

Susceptible host can be anyone.

I - Minimal

II - Moderate

III - High

IV - Extreme

• biosafety levels and their risks

Level I Biosafety

• with agentsnnot known to cause disease in healthy adults

Level II Biosafety

• with agents that are common human pathogens

Level III Biosafety

• with agents that cause serios or lethal disease via inhalation

Level IV Biosafety

• with agents that pose high risk of life-threatening disease

Level I

• biosafety level of B. subtilis

Level I

• biosafety level of M. gordonae

Level I

• biosafety level of S. marcescens

Level I

• biosafety level of K. aerogenes

Level II

• biosafety level of E. coli

Level II

• biosafety level of HIV and HBV

Level II

• biosafety level of Influenza

Level II

• biosafety level of T. gondii

Level II

• Staphylococcus

Level III

• biosafety level of B. anthracis

Level III

• biosafety level of Francisella

Level III

• biosafety level of Brucella

Level III

• biosafety level of M. tuberculosis

Level III

• biosafety level of mold stages of systemic fungi

Level IV

• biosafety level of Ebola

Level IV

• biosafety level of Lassa virus

Standard Microbiological Practices

PPE as needed

• minimum requirements for biosafety level I

BSC I or II

PPE

Autoclave

• minimum requirements of biosafety level II

BSC I or II

PPE

Autoclave

Negative Air flow and Sealed Windows

• minimum requirements for BSL 3

PPE

Autoclave

Negative Air flow and Sealed Windows

BSC

Positive pressure suit

Independent unit with ventilation and waste management

• minimum requirements of BSL4

no or low individual and community risk

• risk group 1

mod individual risk, low community risk

• risk group 2

high individual risk, low community risk

• risk group 3

high individual and community risk

• risk group 4

Risk Group I

• microorganisms unlikely to cause human or animal disease

Risk Group II

• pathogen that can cause disease but unlikely to be a serious hazard to lab workers

Risk Group 3

• pathogen that usually causes serious disease but does not ordinarily spread from one infected individual to another

• there is effective treatment and preventive measures

Risk Group IV

• pathogen that usually causes serious human or animal disease that can be readily transmitted from one individual to another, directly or indirectly

• no available treatment and preventive measures

BSC I

• type of BSC that allow room air to pass into the cabinet and around the area and material within, sterilizing only the air to be exhausted

• only the worker is protected

BSC II

• type of BSC that sterilizes air that flows over the infectious material, as well as air to be exhausted

• worker and sample protected

IIA

A1 - biological samples

A2 - biological samples and treated samples

• type of BSC II where 70% of air is recirculated into the work area

BSC IIB1

• type of BSC II where 30% of air is recirculated and 70% is exhausted

BSC IIB2

• type of BSC II where no air is recirculated

BSC III

• type of BSC that is also known as glove-box cabinet

• most protection to the worker and sample

BSC III

• BSC type with air coming into and going out of the cabinet filter sterilized, and the infectious material within is handled with rubber gloves that are attached and sealed to the cabinet

Sterilization

• destruction of all forms of life including spores

Disinfection

• elimination of a defined scope of organisms

• applied on inanimate objects

Antisepsis

• performed on living tissue but does not kill spores

Bactericidal

• kills bacteria

Bacteriostatic

• inhibits bacterial growth

Prions

Bacterial Spores

Mycobacteria

Nonlipid Viruses

Fungi

Bacteria

Lipid Viruses

• order of lala

Autoclaving

Boiling

Pasteurization

Tyndallization

• methods with moist heat

121 C

15 psi

15-30 mmin

• condition for autoclaving with broth and solid media

132C

15 psi

30-60 min

• conditions for autoclaving for infectious waste

Geobacillus stearothermophilus spores

• indicator used in autoclaving

100C for 10-15 min

• boiling conditions

Pasteurization

• used in the food industru

63C for 30 min

• condition for pasteurization in batch

72C for 15 seconds

• pasteurization condition for flash

140C for 3 seconds

• pasteurization condition for ultra-high temp

Tyndallization

• destroys vegetative cells and spores after three consecutive days of sterilization

Tyndallization

• material is first heated at 100C for 30 minutes through boiling or steaming then placed at RT or at 37C for 24 hours

170C for 2 hours

• condition for hot air sterilization

Bacillus atrophaeus spores

• indicator for hot air sterilization

Incineration

• used in loops and biomedical wastes

Filtration

• method of choice for antibiotic solutions, toxic chemicals, radioisotopes, vaccines, and CHO (all heat sensitive)

0.22 mm

• size of the membranes in filtration method that is used to sterilize air and liquids

0.01 um

• size of membranes in filtration used for retaining some viruses

HEPA Filters

• capable of removing organisms larger than 0.3 um

Ionizing Radiation

• gamma rays or electron beams

Ultraviolet Rays

• nonionizing radiation

bacillus pumilus spores

• indicator for radiation

Alcohol (70-80%)

• denatures proteins and makes lipids soluble

• tuberculocidal

2% Glutaraldehyde

• germicidal in 10 minutes and spricidal in 3-10 hours

• tuberculocidal if used at 25-30C

2% Glutaaldehyde

• sterilizer of choice of not heat-stable medical equipment

1% Sdoium Hypochlorite

• chlorine generally used for disinfection

3-6% H2O2

• active against all vegetative microorganisms, bacterial endospores and fungal spores

Iodine + detergent

• component of iodophor

Iodine + alcohol

• component of tincture

Quaternary Ammonium

• not sporicidal or tuberculocidal

• its use is limited to disinfection of benchtops and floors

1:10 dilution of 5.25% sodium hypochlorite

• recommended by CDC for cleaning blood spills

Ethylene Oxide

• used to sterilize materials that cannot withstand steam

Bacillus

• indicator for Ethylene Oxide

Autoclave

Dry Heat

Filtration

Ionizing Radiation

• sterilization methods

Boiling

Pasteurization

Nonionizing Radiation

Chemical Disinfectants

Antiseptics

• disinfection methods

acute phase before administration of antibiotics

• specimen collection should be done at what phase?

Neisseria

• cotton swab is toxic to

Chlamydia

• wood is toxic to

HSV and SARS-CoV2

• calcium alginate is toxic to

within 30 minutes

• delivery of specimen to the lab after collection of invasive specimens is within ______

2 hours

• delivery of specimen to the lab after collection of most specimen is within ______

Amies

Stuart’s

• examples of holding media

Anaerobe Transport Media

RT

• preservative or transport device for anaerobe specimens

• store temp?

none

RT or 37C

• preservative or transport device for CSF specimens

• store temp?

None

4C

• preservative or transport device for sputum

• store temp?

Cary-Blair (>2 hours delay)

4C (unpreserved)

RT (holding media)

• preservative or transport device for stool for culture

• store temp?

Formalin or PVA

RT

• preservative or transport device for stool for O&P

• store temp?

Boric Acid (>2 hours delay)

4C

• preservative or transport device for urine

• store temp?

none

RT

• preservative or transport device for blood

• store temp?