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Microbiology Mycobacteriaceae NSHS MLT
Microbiology Unit 7.13 Mycobacteriaceae
what is the genus under the family mycobacteriaceae?
how many recognized species of Mycobacterium exist?
up to 100
These are aerobic non spore forming, non encapsulated, slight curved or straigt rods that are sometimes branching, filaments or mycelium-like growth.
Mycobacterium are _____-fast and _____-fast, non motile, not readily stained gram positive rods.
Mycobacterium have a high _____ content of cells and cell walls.
Raidly-growing Mycobacterium require how many days on simple media?
disease-associated species of Mycobacterium require how many weeks on complex media?
how are Mycobacterium incubated?
C in 5-10% CO2 in the dark
This is when cell wall resists decolorization by acidified alcohol.
This describes two or more species, distinction between spp. has no medical importance.
What are the three species in the M. tuberculosis complex?
M bovis/M. bovis (BCG)
This classification system is used for ID of Mycobacterium for public health purposes.
Runyon Classification system
This group of the Runyon classification System is yellow when exposed to light after growing in the dark for >7 days on solid media.
Group I: Photochromogens
What are the two organisms in Group I of the Runyon Classification System?
This group of the Runyon Classification System is yellow when grown in the light OR dark for >7 days on solid media.
Group II: Scotochromogens
What are the two organisms in Group II of the Runyon classification system?
This group of the Runyon Classification System has no pigment in light or dark > 7 days on solid media.
Group III: Nonphotochromogens
What are the two organism in group III of the Runyon classification system?
M. avium complex
What group of the Runyon Classification System takes <7 days to grow on solid media?
Group IV: Rapid growers
What are the three organisms in group IV of the Runyon Classification System?
What is the Non-cultivatable NTM in the Runyon Classification System?
What is the solid Agar based media used for Mycobacterium?
Middlebrook 7H10 and 7H11
What is the Egg based solid media used for Mycobacterium?
Lowenstein Jensen (LJ)
What are the four liquid media used for Mycobacterium?
Mycobacteria growth indicator tube
What are the three rapid methods for diagnosis available for Mycobacterium?
Gas-Liquid and high performance liquid chromatography
What are the four Automated detection systems used for Mycobacterium?
Bactec 460 TB system
Bactec 9000 MB
Bactec MGIT 960
ESP culture system II
What is the automated system used for Mycobacterium that uses radiometric principle and C-labeled substrates.
Bactec 460 TB system
Conventional ID of Mycobacterium is based mainly on what three things?
This is the single best test to ID Mycobacterium tuberculosis?
Niacin Test: +
This Mycobacterium spp. is nitrate (+), urease (+), 68C Catalase (=), and resembles bread crumbs- rough, dry colonies, luxurious growth.
This is a highly contagious disease spread by airbone droplets, inhalation of single viable organism leads to infection.
M. tuberculosis has a high prevalence in patients with ______.
What are three lung infections caused by M. tuberculosis?
Ultimately cavitary disease
What are three symptoms of Pulmonary Tuberculosis?
Specimen for ID of Pulmonary Tuberculosis inculdes ___ first AM sputum samples which should be delivered to the lab within ____ minutes.
What are the following results for M. bovis?
this airborne organism is associated with bovine and human tuberculosis, and contracted via ingestion of contaminated mild, not prevaent due to pasteurization.
What are the following results for M. kansasii?
this organism was originally called "yellow bacillus"
what is the natural reservoir for M. kansasii?
M. kansasii is assocated with what two diseases?
Chronic pulmonary Dx
Extra pulmonary Dx
M. kansasii will cause cervical lymphadenitits and cutaneous dx in what disease?
Extra pulmonary Dx
This organisms is isolated from fresh water and salt water, frequently causes cutaneous infections in fish hadlers and swimming pools.
this Mycobacterium is genitalia normal flora and usually a contaminant.
This mycobacterium is a tap water contaminant.
This ubiquitous Mycobacterium is commonly isolated species in US HIV patients.
M. Avium complex (MAC)
What are the four organisms in the M. avium compex (MAC)?
This Mycobacterium can causes disseminated disease in AIDS patients, and pumonary infections, cervical lymphadenitis, and Disseminated disease in other patients.
M. avium complex (MAC)
This Mycobacterium organism is seen in raw milk, soil, water, and dairy.
Pathogenic and epidemiologic studes of this Mycobacterium are difficult becuase it cannot be grown in vitro.
Diagnosis of this Mycobacterium are made through histopathological examination of skin lesion.
This is an obligate intracellular parasite in man, confined largely to skin (cold surfaces), especially convex and exposed surfaces, testes, peripheral nerves.
leprosy (Hanen's Disease)
M. leprae probably do not grow in ______ organs.
What are the two forms of Leprosy?
Tuberculoid and neural lesions
In what form of Leprosy are organisms rare?
Tuberculoid and neural lesions
In what form of Leprosy are bacilli abundant in tissue, and produce stuffed-cell granulomas.
This is a drapid growing Mycobacterium associated with post operative infection, skin and soft tissue infection, and pulmonary infection.
This is a GP, partially acid fast aerobic actinomycetes that has a beaded appearance on gram stain, is aerobic, and catalase (+).
This organism is associated with mycetoma, lymphocutaneous infections and skin abcess cellulitis in immunocompetent patients.
What is the greatest single problem in mycobacteria recovery?
presence of large numbers of contaminating microorganisms.
Mycobacterium specimens should be refrigerated if not processed within __ hour.
what is used as an expectorant for collection of sputum samples in patients with non-productive cough?
10% NaCl solution
Gastric lavage is collected from ______ and others unable to cooperate with sputum induction procedures.
Gastric lavage collection bottle should contain what?
CSF should be centrifuged for 30 minutes at what speed?
homogenization and decontamination is required for what four specimens?
Tissues (Following grinding)
Other body fluids (except urine)
What type of BSC hood should be used for Mycobacterium?
Laminar flow hood
What are the two decontaminants for Mycobacterium?
Sodium hydroxide (NaOH)
What is used as a digestant for Mycobacterium?
What is the purpose of a digestant in Mycobacterium?
What is used as a neutralization solution for Mycobacterium?
What is the pH of the phosphate buffer nutralization solution for mycobacterium?
how are Mycobacterium specimens incubated?
C at 5-10% CO2
if suspected what two Mycobacterium should be incubated at 25-30
AFB cultures are kept for ___ weeks, examine daily.
This very infectious Mycobacterium can penetrate skin and can survive 5% phenol for 24 hours.
What BSL is required for labs that process mycobacterium?
Cultivating what two Mycobacterium requires BSL 3?
Tuberculin-negative laboratory and hospital personnel are tested _______.
All Mycobacterium procedures and work are performed in what class laminar flow hood with UV light.
Where are centrifuges and shaking machines kept when testing for Mycobacterium?
inside laminar flow hood
Germicidals and disinfectants used around Mycobacterium should aslo be what?
This is designed for rapid diagnosis of organism with high lipid content in cell wall capable of binding fuchsin dye that is not removed by acid alcohol.
Acid-Fast staining procedure
What are the three uses of acid-fast staining procedure.
confirm acid-fast nature
monitor patients on antimycobacterial drugs
determine appropriate dilution for suceptibility testing
What are the three types of acid fast stain?
Ziehl-Neelsen (hot stain)
Kinyoun (cold stain)
Fluorochrome stain (UV microscope)
This stain contains 3g basic fuchsin, 10 ml 90-95% ethanol, and 90 ml 5% aqueous phenol (carbolic acid).
Carbolfuchsin stain (mixture)
What are the two reagents used in stains for mycobacterium?
methylene blue counterstain
this reagent used in Mycobacterium stain contains 3 ml concentrated HCL added slowly to 97ml of ethanol in this order, solution may get hot.
What three reagents are in the Fluorochrome stain?
Fluorochrome stain is viewed using what type of microscope?
these two acid-fast staining techniques are theoretically the same, but the first is more sensitive in detecting lightly staining organisms.
this heat fixed stain should NOT be heated until steaming.
what color are mycobacteria stained in acid-fast staining?
red on light blue background
how many control slides should be made per suspect slide in acid-fast staining?
how should the positve control smear appear for acid-fast staining?
red, small, slightly curved bacilli
possibly beaded and tapered ends
negative acid-fast staining smear should display numerous ______ (non-acid-fast) bacilli.
What is the organism and strain that is used for acid-fast staining quality control?
M. tuberculosis strain H37rv
A minimum of how many fields should be scanned before reporting a negative sample in acid-fast staining?
What is the non-tuberculosis species that appears pleomorphic, usually coccoid in the acid fast stain?
M. avium complex
If large numbers of typical forms of acid-fast bacilli are seen in acid-fast staining it is reasonable to assume they are what organism?
how are Acid-fast bacilli reported in Acid-fast staining?
Positive for acid-fast bacilli
other pathogenic of non-pathogenic mycobacteria or partially acid-fast organisms usually appear as what on the acid-fast stain?
if only one or two acid fast bacilli are seen what should be done?
recommend confirmation with new specimen prior to reporting
if 1-2 organisms are seen in 300 fields using acid-fast staining, what should be reported.
if 1-9 organisms are seen in 100 fields, in acid-fast staining, what should be reported?
if 1-9 organisms are seen in 10 fields, using acid fast staining, what should be reported?
if 1-9 organisms are seen per field using acid fast staining, what should be reported?
if >9 organisms are seen per field in acid fast staining, what should be reported?