Laboratory Safety, Biosafety Protocols, Specimen rejection Flashcards
The lab can be a safe place to work if…
appropriate safety precautions are taken
What is one of the main objectives of the laboratory portion of this course?
to teach and develop those skills that will make the student competent and confident while working in the microbiology laboratory
What microorganisms can be dangerous to your health?
nearly any microbe, given the opportunity and proper conditions
What personal health conditions must be notified to the course coordinator if you have, or develop?
Immunosuppression and allergies.
How is laboratory access restricted?
yes, do not bring friends, family, or pets
only work in the lab as authorized by your instructor- do not perform unauthorized experiments
How must you be dressed in the laboratory? (4 conditions)
- lab coats required, a disposable one is provided at each lab session
- close-toed shoes required (no open-toed or “flip-flops” allowed
- gloves and safety glasses will be provided and must be worn at all times
- tie long hair back
Where should nonessential books and clothing be stored?
in the lockers provided, must provide locks to secure your items
What are the 4 entry procedures?
- put on lab coat
- wash hands with soap and water
- put on gloves and safety glasses
- wipe of table tops with disinfectant
What safety features should you learn of their locations?
- fire extinguisher
- fire alarm
- eyewash
- exits
- first aid kits
- biological spill kits
- emergency phone
What are examples that go against the rule of “put nothing in your mouth while in the lab?”
- using tobacco, chewing gum, eating/drinking in the lab
- pencils, fingers, etc near or in your mouth, eyes, ears, nose
- handling/applying cosmetics or contact lenses in the lab
- artificial fingernails precluding safe handling or lab cultures/equipment
- mouth pipetting
How can you avoid sniffing/smelling your cultures?
keeping the removal of lids from culture plates to a minimum
Are bunsen burners used in this lab?
no
Where should you place your cultures?
in the assigned racks, incubate in your assigned incubator; plates can be attached to the racks before placing them in the incubator
Would there ever be a reason to take any cultures out of the lab?
NO, it is forbidden to remove cultures from the lab
Where should contaminated or used material be disposed?
Should be disposed quickly in the red biohazard bags. What must be done to the material containing bacteria before disposal?
What are the 3 disposal sites?
- biohazard bags
- sharps container
- plastic tubs
What items are disposed in each?
Must a minor accident be reported?
Any accident (potential exposure to an agent, spills, or broken glass or cuts), no matter how minor, must be reported to the instructor immediately
What is the procedure for handling infectious spills?
- notify instructor
- allow aerosols to settle
- put on protective equipment (like gloves)
- soak paper towels with disinfectant and cover spill with paper towels
- discard paper towels in biohazard bag
What is the procedure for handling broken glass?
- notify instructor
- put on protective equipment (gloves)
- use broom and dust pan to clean up broken glass (do NOT use bare hands)
- put broken glass in sharps container (do NOT put it in the trash can!!)
- seek immediate aid from instructor if you cut or injure yourself in any way, no matter how minor the event
What is the procedure for handling chemical spills?
notify instructor immediately
What are the 4 exit procedures?
- wipe off your table tops with disinfectant
- remove your gloves, remove safety glasses and return them to supply drawer at your seat, remove lab coat and place it in large red biohazard bag
- wash hands with soap and water
- gather your lab notebook and leave the lab IMMEDIATELY
The first violation of the laboratory safety rules results in:
- immediate dismissal from the lab
- loss of credit for the lab period
how could you determine if you are contaminating your house?
take a sample and test is
three contaminates of an experiment
1.air
2.bench
3.you
all solid non-contaiminated waste
go into regular trash
all small solid contaminated waste which includes….
-pipette tips and eppendorf tubes
they go in the trash jars on each table, when full empty them into the to be autoclaved trash
large solid contaminated waste
-Kim wipes, paper towels, gloves, Parafilm
they go into the to be autoclaved trash
when to be autoclaved trash is full you should….
put it in the sink by the autoclave and place an new bag in the rash can (found on top of autoclave)
microscope slides
if not scratched they need to be cleaned with 20% bleach for 5-10 min washed with soap and water, and allowed to dry. When dry put them back in drawer underneath the hot water bath
plastic seriological pipettes
contaminated- goes into the large sharps contain (take of wrapper!(trash or autoclaved trash)
non-contaminated-regular trash
all pasteur pipettes
go into the sharps container
liquid cell waste
treat with 20% bleach for 5-10 min with cap ON, pour into cell waste flask, another rinse with 20% bleach, wash with soap and water. Put test tubes on correct rack with cap. take off tape and wash off labels
solid bacterial cultures
needs to be put on the test tube rack labeled to be autoclaved, if full label a new one
biohazard spills
put paper towel over to minimize aerosols, top with 20% bleach, sit for 10-15, scoop up mess, wipe surface with bleach or 70% ethanol
CDC
Center for Disease Control and Prevention
NIH
National Institute of Health
BSL
stands for Biosafety level
biosafety 1
-not known to consistently cause healthy adults harm
-wash hands a lot
-no eating or drinking, makeup
-not storage of food
-no mouth pipetting
-use container much larger than volume (1/3 size)
-decontaminate surfaces with ethanol
-lockable doors
-gloves and coats recommended
-personal belongings in seperate room recommended
examples of BSL 1
-escherichia coli K12
-candida albicans
-infectious canine hepatitis virus
BSL2
-associated with human disease and varying severity
-moderate risk
-treatment and vaccines are available
-not trasnferrable through air
-all BSL 1 rules
-appropriate vaccines
-personal belongings must be seperate
-lab coat and gloves required
-must have autoclave
-need to show BSL 1 competancy
examples of BSL 2
-measles
-salmonella
-hepatitis B virus
BSL 3
-airborn but there is treatment
-ex: TB
BSL 4
-not all are airborn
-little to no treatment- exotic
-ebola
closest first aid kit?
in the back
nearest telephone
cell phone or around the corner
why use 70% vs 95% ethanol
would evaporate too quickly, must stay in contact long enough with the bacteria to kill the microorganisms it takes time!
chains of infection steps
1) etiologic agent
2)reservoirs
3) portal of exit
4) mode of transmission
5) portal of entry
6) susceptible host
etiologic agents (6)
1) bacteria
2)viruses
3) fungi
4) parasitic worm
5) protozoa
6) prions
bacteria example
mycrobacterium tuberculosis, TB–lung infection
virus example
influenza virus-flu–respiratory illness
fungi example
candida albicans–thrush
parasitic worms example
trachinella spiralis, trichinellosis–transmused by undercooked meat
protozoa example
plasmodium spp., malaria–transmused by infected mosquitoes
prions example
scrapie
reservoir list (3)
1)human
individual that are sick, carriers who are not
2)animal
3)environment
human individuals who are sick example
rhino virus-common cold
human carriers example
hepatitis A virus, hepatitis A–50% do not have symptoms
animal example
yersinia pestis, bubonic plague–rats
environment example
clostridium tetani, tetanus, rusty nail
portal of exit list (5)
1)respiratory
2)genitourinary
3)alimentary (mouth through anus)
4)skin
5)transplacental
respiratory example
rhino virus-common cold
genitourinary
neisseria gonorrhoeae-gonorrhea
alimentary
hep A virus-hep A
skin
small pox virus-small pox
transplacental
human immune deficiency virus-aids
mode of transmission list (2)
1) direct- occurs immediately due to direct contact with a human animal or environmental reservoir. This includes droplet spreading
2) indirect- use of vectors (like mosquitoes) or through inanimate meachanisms
direct example
Shigella sonnei, shigellosis
indirect use of vectors
pasmodium spp, malaria-mosquiotoes, ticks, and fleas
indirect inanimate mechanisms
escherichia coli o157–E coli
portal of entry (5)
1)respiratory track (inhalation)
2) alimentary tract (ingestion)
3) broken skin (penetration)
4) genital membranes
5) ocularmucosa (mucous membrane of eye) not a means of escape
susceptible host list (3)
1) genetic factors
2) general resistance factors
3) specific acquired immunity
active immunity and passive immunity
general resistance factors
include our intact skin and mucus membranes-skin and staph infections
specific acquired immunity example (active)
is achieved by experiencing an infection–chicken pox
specific acquired immunity example (passive)
is transferred of antibodies through breast milk
exposure control (7)
1)universal precautions (know what you are doing)
2)immunizations (vaccines are up to date)
3) good personal work habits (wash hands, no eating in lab, wear closed toed shoes)
4)containment (lab has lockable doors, minimize aerosols, clean up spills)
5)personal protective equipment (gloves and goggles)
6)waste disposal (correct container, sharps broken glass)
7) emergency procedures for accidental exposures (call 911, notify the in structure)
1.List general guidelines for proper specimen collection from a suspected site of infection.
Collection of specimen, sterile container and proper transport media
- Which type of swabs may contain fatty acids that inhibit some strains of fastidious bacteria?
Cotton Swabs
3.What type of swabs are suitable for collection of a specimen for culture of Neisseria gonorrhoeae?
Dacron or rayon swabs
4.What can be used as a preservative for stool specimens for culture?
formalin and PVA
- What can be used as a preservative for urine specimens?
hydrochloric acid, boric acid, acetic acid and toluene.
6.How should specimens for microbiology be transported?
Deliver all specimens as soon as possible to the Microbiology Laboratory
7.What information is required on microbiology requisitions?
Patient’s full name (last, first)
Patient’s medical record number (if available)
Patient’s location
Patient’s date of birth and sex
Physician’s full name
Test(s) requested: Check the box to the left of the test on the requisition. For tests not listed on the form legibly, print name of ordered test on the blank lines.
Diagnosis and/or ICD-9 code
Date and time of specimen collection
8.List several causes for specimen rejection in the microbiology laboratory.
Container not identified
Container and request form have different ID’s
Insufficient information marked on form
Specimen grossly contaminated
Specimen submitted in improper container
Excessive delay between specimen collection and arrival in laboratory
Inadequate specimen for number of tests requested
Some factor renders specimen inadequate for request.
Examples: sputum on a swab or in tissue paper, stool for ova and parasites with gross barium, visible; specimen placed into formalin; container broke or leaked during transport
9.As a rule, in what situation should every effort be made not to reject a specimen?
For CSF samples or neonatal samples
10.Which specimens may be refrigerated?
Urine, respiratory, stool
11.Which specimens should never be refrigerated?
CSF
12.Which microorganisms are particularly susceptible to adverse environmental conditions such as atmosphere, temperature, and pH?
Niessirrea gonnorrhea, legionella
13.When is the ideal time to obtain a blood culture?
Morning or early stage of infection
14.How many blood cultures should be drawn?
4 total 2 aerobic and 2 anarobic
15.What volume of blood should be drawn for blood cultures?
30 ml
16.Which anticoagulant is used for blood cultures and why?
sodium polyanetholesulfonate
17.What ratio of blood to broth is recommended for blood cultures?
1:10
- Which tube of CSF should be sent to the microbiology lab for culture and sensitivity and why?
Second container
Match the medium with its classification (some may be used more than once)
1. Modified Thayer-Martin agar
e. antibiotic, selects for pathogenic Neisseria
Match the medium with its classification (some may be used more than once)
- Colistin-nalidixic acid agar
antibiotic, selects for gram-positive bacteria
Match the medium with its classification
3. MacConkey agar
c. selective for fecal pathogens
Match the medium with its classification
5. Blood agar
enriched, nonselective
Match the medium with its classification
6. Eosin-methylene blue agar
selective for fecal pathogens
Complete hemolysis of blood is known as:
B-hemolysis
The specimen of choice for bacterial culture of urine is
Clean-catch midstream
A throat swab is submitted for anaerobic culture. This specimen should be:
rejected
A spinal fluid specimen is submitted for Gram stain and culture during an afternoon shift. The physician also requests that an aliquot be saved for possible serological studies, which are performed only on day shift. The most correct action is:
Inoculate culture, perform a Gram stain, and refrigerate remaining spinal fluid
The general guidelines for collection of blood cultures state:
two to three sets per 24 hours are usually sufficient to diagnose bacteremia
Prompt delivery of specimens for microbiology is essential for accurate culture workup. Which of the following is NOT A consequence of delay in specimen delivery?
normal flora may no longer be isolated
Which of the following is NOT considered to be a sterile site?
oral cavity
An abdominal abscess specimen received in the laboratory produced abundant growth when grown anaerobically but produced no growth when the culture plates were incubated aerobically. This culture contains:
an obligate anaerobe
The optimal time to collect blood cultures is:
while the fever spikes
Most pathogenic bacteria prefer an incubation temperature of:
35 - 37 degrees C
Those bacteria that prefer growth under increased CO2 are known as:
Capnophilic
Which of the following should be monitored as part of the quality control program in the microbiology laboratory?
a. temperature of incubators and refrigerators
b. plating media with known positive and negative controls
c. oxygen content and CO2 content of incubators
d. all the above
d. all the above
The catalase test is useful in differentiation of:
differentiation of Staphylococcus from Streptococcus