INTRODUCTION TO MICROBIOLOGY (BACTE LAB MOD1)

Biosafety Program

Its fundamental objective is to contain any potentially hazardous biologic agent or toxin

Universal Precaution

Applied to any blood and body fluids containing visible blood

HIV epidemic

Universal Precaution is intended in response to?

True

TRUE OR FALSE. Universal Precaution is followed if patient has known blood borne disorder or if the patient has unknown infection status

FALSE

TRUE OR FALSE. : Airborne pathogens are addressed in universal precaution.

TRUE

TRUE OR FALSE. In order to address the pathogens that can be transmitted airborne – we refer to the standard precaution.

Standard Precaution

Treat patient’s blood and body fluids as potentially hazardous

TRUE

TRUE OR FALSE. Standard precaution is followed regardless of the patient’s infection status.

Handwashing – Gown – Mask – Goggles/Face shield – Gloves

Enumerate the process of donning PPE

Gloves – Goggles/Face shield – Gown – Mask – Handwashing

Enumerate the process of doffing PPE

Use of PPE

Overlap of the Standard and Universal Precaution

Handwashing

most effective way to stop the chain of infection

Biologic Safety Cabinet

A device that encloses a workspace

Filtration

BSC sterilizes air that contains infectious material thru

High Efficiency Particulate Air (HEPA) Filter

filters air, can remove objects that are larger than 0.03 um

Class I

Least effective of all BSC classes

Class I

Entirely open BSC

Laminar Flow

BSC II is also known as?

Class II

BSC that used in most laboratories

Class II A

BSC that exhaust air inside the room

Class II B

BSC that exhaust air outside the room

Class III

BSC that is most effective of all classes, entirely closed, has a part where objects may pass through and has built-in gloves, sealed to the cabinet

Practice of Standard Laboratory Techniques

Precaution of BSL-1

Minimal Risk

Description for BSL-1

Bacillus subtilis, Mycobaterium gordonae

Examples of BSL-1 organisms

Moderate Risk

Description of BSL-2

Use of PPE, Use of Biohazard warning signs, Decontamination of infectious wastes

Precaution for BSL-2

Salmonella, Shigella, S. aureus, HBV, HIV

Examples of BSL-2 organisms

High Risk

Description for BSL-3

Use of PPE, Laboratory must have suitable airflow

Precaution for BSL-3

Systemic mycoses agent, M. tuberculosis, Coxiella burnetti, Brucella, Franciscella, St. Louis Encephalitis virus

Examples of BSL-3 organisms

Extreme Risk

Description for BSL-4

Use of Full Body PPE, Laboratory must be situated in a separate building

Precaution for BSL-4

Arenavirus, Margburg virus

Examples of BSL-4 organisms

BSC I

What class of BSC does BSL-1 organisms use?

ingestion, mucus membrane, and percutaneous exposure

How are BSL-2 organisms acquired?

BSL-2 organisms

Which BSL organisms are associated with laboratory acquired infections?

BSC II

Which BSC is used in BSL-2 organisms?

BSL-3 organisms

What organisms impose high risk infection but has existing treatment?

inhalation or aerosol

BSL-3 organisms are acquired through?

BSC II

BSL-3 organisms uses which BSC?

BSL-4 organisms

Which organisms impose high risk infection, has no treatment and can cause life-threatening diseases?

Highly infectious; Smallpox virus, Franciscella spp., Bacillus anthracis

BIOTERRORISM CATEGORIES. Description and examples for Category A

Moderate morbidity, Low mortality; Rickettsia spp., Coxiella spp., Burkholderia pseudomallei

BIOTERRORISM CATEGORIES. Description and examples for Category B

Emerging pathogens; Yellow Fever Virus, Dengue Fever, Ebola Virus, MERS-COV, Zika Virus

BIOTERRORISM CATEGORIES. Description and examples for Category C

Yersinia pestis

Bioterrorism agent used in Black Plague

Family Coronaviridae

MERS, SARS, COVID-19 are under family?

Sterile Specimen

Type of specimen that is an aspirated fluid, can be processed immediately

Non-sterile specimen

Type of specimen that involves processing prior to actual bacteriologic study

Urine, stool, sputum

Examples of non-sterile specimen

Before Antibiotic Therapy, Aseptic Collection, Quantity Sufficient, Acute Stage, Prompt Delivery, Proper Sterile Delivery

6 general rules for specimen processing

TRUE

TRUE OR FALSE. One of the main rule that we follow in bacteriology is that all equipment that we use must be sterile.

True

TRUE OR FALSE. Presence of contaminant may cause confusion in identifying the actual bacteria present in the specimen.

Bacteremia, Sepsis, Fever of Unknown Origin

What are detected in a blood culture specimen?

Bacteremia

It is the presence of viable bacteria in the blood

Coagulase Negative Staphylococcus: S. epidermidis, S. saprophyticus; E. coli, P. aeruginosa, S. aureus, Bacteroides fragilis

Causes of Bacteremia

Septicemia

It is the type of bacteriemia accompanied by systemic inflammatory response

70-95% alcohol – iodine scrub/chlorhexidine – alcohol rinse, 70% alcohol and chlorhexidine/iodophor

2 Antiseptic techniques for blood culture collection

S. epidermidis, viridans streptocci, Propionibacterium acnes, Bacillus spp., and diptheroids

Common contaminants in blood culture specimen

Cutibacterium acnes

What is the new name for Propionibacterium acnes?

S. epidermidis

Normal flora of the skin

Bacillus anthracis

Bacillus species are common contaminants in a blood culture specimen, except for?

0.025% SPS

Anticoagulant used in blood culture collection?

Prevents phagocytosis, Prevents compliment fixation and Can neutralize some antibodies

3 Mechanisms of SPS as used in a blood culture include?

compliment fixation

a series of activation that happens in the body in which the result is Membrane Attack Complex (MAC)

Membrane Attack Complex (MAC)

causes cell lysis.

Neisseria, Peptostreptococcus anaerobius, Garnderella vaginalis, Streptobacillus moniliformis

SPS is inhibitory for which organisms?

Add 0.1% Gelatin

Remedy for SPS, to neutralize its effects

Sodium amylosulfate, 0.5-1.0% Sodium Citrate, and Heparin

Other additive for blood culture collection include?

Heparin

This additive is most likely used to recover virus in the blood

Some gram negative bacteria

Heparin can inhibit the growth of?

Fungi; yeast

Heparin is not suitable for the recovery of ___ because it can inhibit the growth of ____?

EDTA

It is never used for blood culture (anticoagulant)

TSB, BHI, Brucella broth

Culture medias used in Blood culture?

1:10

Blood to Broth ratio

hemolysis, pellicle, or turbidity

Indications for positive growth in the broth includes the presence of?

Blood Agar Plate, Chocolate Agar Plate, and MacConkey Agar

Three subculture medias used in blood culture?

FALSE

TRUE OR FALSE. After negative growth in the broth, we do not need to subculture in Blood Agar Plate, Chocolate Agar Plate, And MacConkey Agar

Pathogenic

Considerations in Sub cultured media. Positive growth within 1- 3 days, indicates what type of bacteria?

Contaminant

Considerations in Sub cultured media. Positive growth in the 5 th day indicates what?

Sputum

This specimen is used to detect respiratory tract infections such as tuberculosis or pneumonia

Mycobacterium tuberculosis

Sputum spx is assumed to be containing?

Klebsiella pneumoniae, Staphylococcus aureus, Pseudomonas aeruginosa (px with cystic fibrosis)

What are the agents of pneumonia?

Streptococcus pneumoniae, Haemophilus influenzae, Staphylococcus aureus, Klebsiella, Enterobacter, Serratia

What are the agents for ventilator-associated pneumonia?

BAP, MAC, Lowenstein-Jensen/Middlebrook 7H-10 7H-9 broth

Culture media used in sputum culture?

Lowenstein-Jensen/Middlebrook 7H-10 7H-9 broth

Culture media for the detection of M. tuberculosis?

NOT PRESENT

When incubated at 37 deg C at LJ media, and there is no growth within 8 weeks, is M. tuberculosis PRESENT OR NOT?

PRESENT

When incubated at 37 deg C at LJ media, and there is growth within 2-3 weeks, is M. tuberculosis PRESENT OR NOT?

Cauliflower-like appearance

Colonies of M. tuberculosis in LJ Media appears what?

Gentamicin Blood Agar Plate

This culture media is used to recover S. pneumoniae

Bacitracin Chocolate Agar Plate

This culture media is used to recover H. influenzae

FALSE

TRUE OR FALSE. Sputum is a sterile specimen, thus, its quality may not be evaluated and we should directly proceed in processing it.

Bartlett’s Classification

It is used to determine if sample is sputum or saliva

Sputum

Which specimen indicates less than 10 SEC; more than 25 PMN?

Saliva

Which specimen indicates more than 10 SEC; less than 25 PMN?

Alveolar macrophage and columnar cells

Indication of poor collection in sputum

4 deg Celsius (refrigerator temperature)

Storage temp of Sputum if not processed immediately

deep cough

Sputum is collection through?

5-10 ml

Volume of Sputum

Digestion

Process of liquefying mucous by using digestant/mucolytic