MODULE 1 PART 2 Flashcards
Nutritional req: light as energy source
Phototrophic
Nutritional req: chemical compounds as energy source, most bacteria are this
Chemotrophic
This uses inorganic compounds as carbon source for growth
Autotrophic/ Lithotroph
This uses organic compounds as carbon source for growth
Heterotrophic/Organotroph
Example of organic compounds that bacteria uses as carbon source
Glucose
This requires oxygen for growth
Obligate aerobe
How many percent of oxygen needed in Obligate Aerobe?
21%
What are the organisms included in Obligate Aerobe? (MPBBm)
Mycobacterium, Pseudomonas, Bacillus, Brucella micrococcus
Cannot grow in the presence of O2, they die in the presence of oxygen
Obligate Anaerobe
Example of organisms in Obligate Anaerobe (B and C)
Bacteriodes and Clostridium
This is used to ensure anaerobiosis in the laboratory
Gaspak jar
Can live with the presence or absence of oxygen O2
Facultative Anaerobe
Most bacteria are this kind of oxygen requirement
Facultative Anaerobe
Example of organisms under Facultative Anaerobe
Staphylococcus, Streptococcus, E. coli, Salmonella
This can grow in the presence of O2 but it grows best in an anaerobic environment
Aerotolerant
Example of organism in Aerotolerant
Lactobacillusb
This can grow in reduced level of O2, requires small amount of oxygen/air
Microaerophilic
What is the requirement for microaerophilic?
- 5 to 10% O2
- 10% CO2
- 85% N
What is the requirement for obligate anaerobe?
- 0% O2
- 5% CO2
- 10% H
- 85% N
Most bacteria grow well at 7.0-7.5 ; most bacteria are this
Neutrophile
Grows at an acidic pH
Acidophile
Organism that survives in the pH = 3.0
Lactobacillus
Organism that survives in the pH = 10.5
Vibrio cholerae
0-20° C; cold-loving
Psychrophilic
Temperature of Mesophilic
Moderate ; 20-45° C
50-125° C; Heat-loving
Thermophilic
The bacteria is not growing well in the presence of high temperature, the growth is not okay
Thermoduric
Incubation at 4° C for several weeks
Cold enrichment
2 organisms that can tolerate Cold Enrichment
- Listeria monocytogenes
2. Yersinia enterocolitica
These are organisms with short optimum temperature range
Stenothemophilic
These are organisms with wide temperature range
Eurithermophilic
This is the temperature at which an organism grows best and every organism has its own
Optimum temperature
Motility is best detected at ____
Room temp
This requires concentrations of 5% to 10% CO2 for optimal growth, this needs high CO2
Capnophilic
Example of capnophilic organisms
Streptococcus and Neisseria
This is used to supply with increased CO2 and this is for capnophiles
Candle Jar
Gaspak jar uses _________ to prevent O2
Palladium catalyst
3 Indicators used in Gaspak Jar
- Colorless in the absence of O2
- Methylene blue (becomes blue when there’s O2)
- Resazurin (becomes pink when there’s O2)
This requires high salt concentration or increase in salt
Halophilic
Species that are known to be microaerophilic and capnophilic
Campylobacter spp.
7.5% salt is tolerated by
Staphylococcus aureus
6.5% salt is tolerated by
Enterococci
8-10% salt is tolerated by
Vibrio
This is the phase where there is an increase in size but the growth rate still remains at 0%
Lag phase/ Adjustment phase
This is the phase where the cell division starts thus growth rate increases, rapid exponential growth, and where bacteria become susceptible to antibiotics
Log phase
This is the phase where the nutrients are used up, toxic product accumulate, sporulation begins and the dead cells is equal to the living cells
Stationary phase/ Plateau
This is known to be the bacterial death of cells where complete stoppage of bacterial growth or cessation happens, where there is development of unfavorable pH
Death phase/ Stage of decline
What are the two causes of bacterial death?
- Development of unfavorable pH
- Accumulation of toxic waste
What are the factors under manual identification? (SSAMSC)
Size, Shape, Arrangement, Motility, Staining characteristics, Cultural characteristics
What are the three ways of identifying bacteria? (MSA)
Manual, Semi-automated through API, Automated thru VITEK, Maldi Tof
API stands for
Analytical Profile Index
This method is not usually utilized nowadays. Half is done by med tech, half is done by machine
Semi-automated thru API
This method is prone to error, solely done by med tech
Manual
This is an automation program for bacterial identification and susceptibility
VITEK
These are the 2 commonly used types of staining
Gram staining, Acid Fast staining
This is part of the manual identification where the use of selective media and biochemical tests are utilized
Cultural characteristics
Most reliable or faster method of bacterial identification
Automated thru VITEK, Maldi Tof
This is the process of artificially coloring the organism with dyes/stains
Staining
Dye’s color is also the resulting color
Simple staining
What is the stain used in simple staining?
Methylene blue
In this staining technique, the background is stained not the organism thus the organism will appear colorless
Indirect/Relief/Negative
What are the stains used in Indirect/Relief/Negative staining?
India ink and Nigrosin
This staining demonstrates special features of cell. The desired part will only be stained.
Special staining
Capsular stains (HA)
Hiss, Anthony’s
Spore stains (DSFWc)
Dorner’s, Schaeffer and Fulton; Wirtz conklin
Counterstain of Schaeffer and Fulton
Safranin: stains other structures to emphasize presence of spores
Flagellar stains (GFCL)
Gray’s, Fisher & Comm, Leifson
Metachromatic granules stains (ANLPMb)
Albert’s, Neisser, Ljubinsky, Ponder, Methylene blue
This staining is used to differentiate one organism from another such as gram staining and acid fast staining
Differential staining
In gram staining, which is the primary dye?
Crystal violet
What is the mordant in gram staining?
Gram’s Iodine
The decolorizer in gram staining are?
Alcohol, acetone, alcohol acetone mixture
The counterstain in gram staining is?
Safranin
The gram (+) color is …
Purple
The gram (-) color is …
Red/pink
Enumerate the staining procedure in order (SIMICSHSM)
Stool, inoculate, Media (petridish), incubate, colonies, slide, heat fixation, staining, microscope focus
All are gram (+) cocci except… (NVB)
Neisseria, Veillonella, Branhamella
All are gram (-) bacilli except… (MCCBELL)
Mycobacterium, Clostridium, Corynebacterium,Bacillus, Erysipelothrix, Listeria, and Lactobacillus
Higher forms of organisms like _______ are gram (+)
actinomyces, streptomyces, yeast and molds
All spiral organisms are reported as gram ___
Gram (-)
Not Gram stained organisms are… (RCMU)
Rickettsiae, chlamydia, mycoplasma, ureaplasma (they dont have cell wall and intracellular)
These are difficult to stain but once stained, they are difficult to decolorize because of mycolic acid and hydroxymethyl acid
Acid fast organisms
Wetting agent in Acid Fast Staining
Tergitol
2 common methods of acid fast staining
- Ziehl Neelsen Method
- Kinyoun’s Method
This is known to be the hot method, best method for Direct Sputum Smear Microscopy which detects acid-fast bacilli (AFB)
Ziehl Neelsen Method
Give the Primary dye, Mordant, Decolorizer, and counterstain of Ziehl Neelsen Method
- Primary dye: Carbol Fuchsin
- Mordant: Steam/Heat
- Decolorizer: 3% acid alcohol (HCl + ethanol)
- Counterstain: Methylene blue or Malachite green
This method is known as the cold method and best stain for AFO in tissues. This has a greater concentration of phenol and basic fuchsin and does not require heating in order to stain properly
Kinyoun’s Method
Give the Primary dye, Mordant, Decolorizer, and counterstain of Kinyoun’s method
- Primary dye: Carbol Fuchsin
- Mordant: Wetting agent (tergitol)
- Decolorizer: 3% acid alcohol (HCl + ethanol)
- Counterstain: Methylene blue or Malachite green
This will take color of the primary dye ; red against a blue background
AFO = Acid Fast Organism
This will take the color of the counterstain ; blue/green
NAFO = Non Acid Fast Organism
This is other method of acid fast staining that is used to differentiate M. smegmatis (blue) from M. tuberculosis (red)
Pappenheim
This is other method of acid fast staining that is used to differentiate M. leprae (red) from M. tuberculosis (blue)
Baumgartens
This is other method of acid fast staining that is used for M. leprae, uses hematoxylin as counterstain instead of methylene blue
Fite Faraco
This is known to be the most sensitive acid fast staining
Fluorochrome staining
Give the Primary dye, Mordant, Decolorizer, and counterstain of Fluorochrome Staining
- Primary dye: Auramine-rhodamine dye
- Mordant: No mordant
- Decolorizer: 0.5% acid alcohol
- Counterstain: 0.5% Potassium permanganate
The result of Fluorochrome staining will be….
AFO will appear yellow against black background
Type of culture media as to Physical State: this type of media has NO solidifying agent, 0% agar, placed in a tube (deep or broth)
Liquid
_______ in liquid media is an indication of growth
Turbidity
Examples of liquid media are… (TSB, APW, BHI)
Trypticase Soy Broth, Alkaline Peptone Water, Brain Heart Infusion
Type of culture media as to Physical State: this is place in a tube to produce slant/deep/butt manner with 0.5-1% agar.
Semi-solid
Example of semi-solid media
SIM media
Type of culture media as to Physical State: can be placed either on a a petri dish or a tube
Solid
Type of media that is initially liquid, will solidify, but dan be liquefied again by heating
Solid Liquefiable
Examples of Solid Liquefiable Media (EM, MSA)
EMB, MacConkey, Mannitol Salt Agar
This is a type of media that will no longer liquefy despite being subjected to heat
Solid Non-Liquefiable
Example of Solid non liquefiable
Rice Medium
______ developing in a plated media indicates bacterial growth
Colonies
This is type of media as to Physical state for both solid and liquid
Biphasic
In Biphasic, Castaneda is used for _____
Brucella
In Biphasic, HBT is used for _____
Gardnerella vaginalis
Type of culture media as to Purpose: This contains only the necessary nutrition to support bacterial growth for routine cultivation of bacteria, no supplements/vitamins added
General purpose/general isolation media
Example of general purpose media
Nutrient agar/nutrient broth
Type of culture media as to Purpose: This is used to enhance bacterial growth or increase bacterial yield, usually contains supplements and vitamins
Enrichment media
Selenite broth =
Salmonella and Shigella spp.
Tetrathionate broth =
Salmonella and Proteus
These are organisms unable to grow in ordinary media
Fastidious organisms
Type of culture media as to Purpose: this is used to support growth of fastidious organisms and it usually contains blood
Enriched media
This blood agar is used for streptococci
Sheep BA
This blood agar is used first choice for Haemophilus
Horse BA
This blood agar is used for Gardnerella vaginalis
Human BA
This blood agar is prepared by heating blood agar and used for Neisseria
Chocolate Agar Plate (CAP)
This type of culture media as to purpose promotes growth of the desired organism while inhibiting the growth of others because it contains inhibitors
Selective media
Complete the ff (even the acronym):
CTBA is for _____ and the inhibitor is _____
Cystine Tellurite Blood Agar is for C. diphtheriae and the inhibitor is potassium tellurite
Complete the ff (even the acronym):
LJM is for _____ and the inhibitor is
Lowenstein Jensen Media is for M. tuberculosis and the inhibitor is malachite green
Complete the ff (even the acronym):
TMM is for _____ and the inhibitor is _____
Thayer Martin Media is for Neisseria spp. and the inhibitor is antibiotics
Complete the ff (even the acronym):
MCA is for _____ and the inhibitor is _____
Mac Conkey Agar is for Gram (-) bacilli and the inhibitor is crystal violet
This type of culture media as to purpose is used to differentiate organisms growing together
Differential media
This is used to differentiate gram negative bacilli as to lactose fermenters or non lactose fermenters
EMB (Eosin-Methylene-Blue)
This is used to differentiate vibrio as to sucrose or non-sucrose fermenters
TCBS (Thiosulfate-citrate-bile-salts-sucrose)
This type of culture media as to purpose is under 2 categories (dala dalawa) example is EMB and Mac Conkey Agar
Selective and Differential Media
This type of culture media as to purpose maintains viability of organisms or keep the slecimens alive during the transport
Transport media
This is transport media specifically for Neisseria
JEMBEC (John E Martin Biological Environmental Chamber)
This is transport media specifically for stool pathogens like Vibrio
Cary Blair
This is transport media specifically for viral transport media
Media for Antibiotic Susceptibility Testing example is
MHA (Mueller Hinton Agar)
This is a culture media as to purpose used in the performance of biochemical tests done in the lab to facilitate species.
Biochemical test media
TSI, LIA, Urea agar, SIM, Simmon citrate Agar, MRVP broth are examples of
Biochemical test Media
This is a culture media used to detect organisms motility
Motility medium
2 Examples of motility medium
SIM and MIO
This is a culture media as to purpose used to support growth of anaerobes
Anaerobic media
Chopped meat media and Thioglycollate are examples of
Anaerobic media