Bacterial Cultivation Flashcards
These are bacteria that get their energy from the light.
Phototrophs
Acid loving bacteria
Acidophilic
Examples of Strict/Obligate Aerobes
- Mycobacterium
- Micrococcus
- Pseudomonas
- Neisseria
- Brucella
- Francisella
- Bordetella
- Leptospira
What does Lactose yield when it is fermented?
Acid (low pH)
MSA Agar
- Original Color:
Light or Salmon Pink
An agar plate that has intact RBC.
Blood Agar Plate (BAP)
It is typically used for decontamination.
NaOH
These are bacteria that obtain energy by breaking down chemical compounds in their environment.
Chemotrophs
Which Stage of Bacterial Growth?
- There is a sudden increase of bacteria because of rapid generation or doubling time.
Logarithmic Phase
MAC Agar
- Indicator:
Neutral Red
The number of generation per hour
Growth Rate Constant
MSA Agar
Mannitol fermenters will appear _________.
Yellow
Together, these other sources make up _____% of the dry weight of the bacteria.
4
SSA Agar
- Sulfur Source:
Sodium Thiosulfate
EMG AGAR
Non-lactose fermenters will appear _________.
Colorless
Tissue Culture Media: HELA Cell Line
Source:
Human Cervical Carcinoma
TCBS Agar:
- Inhibitors:
Thiosulfate, citrate, bile salts,
Most pathogenic bacteria can grow in a _________ or _________ environment.
Neutral or Slightly Alkaline
GC-LECT Agar
Proteus Inhibitor:
Trimethoprim
Dye Inhibitors for Gram Positive Bacteria
- Crystal Violet
- Eosin
- Methylene Blue
- Brilliant Green
What are examples of Basic Culture Media?
Nutrient Agar, Nutrient Broth, TSA, TSB
GC-LECT Agar
Fungal Inhibitor:
Amphotericin B
CAP contains which factor/s?
X and V
These are bacteria that require special or additional requirements to grow in culture media.
Fastidious Bacteria
What are the 3 Major Nutritional Requirements for Bacterial Growth?
- Carbon
- Nitrogen
- Energy
It is used for the determination of bacterial pathogen’s ability to decarboxylate or deaminate lysine.
Lysine Iron Agar (LIA)
Modified Thayer Martin Agar
Gram Negative Inhibitor:
Colistin
Bacterial metabolism with the use of oxygen.
Oxidation
GC-LECT Agar
Gram Positive Inhibitor:
Vancomycin
The growth of microorganisms
Culture (noun)
Martin-Lewis Agar
Proteus Inhibitor:
Trimethoprim
EMG Agar
- Selective for:
Gram negative enteric bacilli
Alkali loving bacteria
Alkaliphilic
Tissue Culture Media: A549 Cells
Source:
Human Lung Carcinoma
Antibiotic Inhibitors for Fungi
- Nystatin
- Anisomycin
- Amphotericin B.
Enumerate the 4 Stages of Bacterial Growth
- Lag Phase
- Logarithmic Phase
- Stationary Phase
- Death Phase
This is also used as a base medium in the preparation of other culture media.
Gen.Purpose/Primary/Basic/Basal/Supportive/General Isolation Culture Media
New York City Agar
Proteus Inhibitor:
Trimethoprim
Which Stage of Bacterial Growth?
- Oxygen becomes limited
Stationary Phase
Martin-Lewis Agar
Gram Positive Inhibitor:
Vancomycin
It is usually contained in a container that can be made of glass or disposable plastic.
Plated
Non-Lactose Fermenters
- ALL Salmonella except arizonae
- ALL Shigella except sonnei
- ALL Yersinia except enterocolitica
- Proteus
- Prudencia
- Morganella
- Edwarsiella
An artificial preparation in the laboratory which contains basic foundation of nutrients and a solidifying agent to support the growth of microorganisms.
Culture Media
The standard size in diameter of a plated media.
100 mm
SSA Agar
- Salmonella Result:
Colorless, w/ H2S and w/ blackening
Eurithermophilic
Wide range of temperature
A slow-growing bacteria is ___________.
Mycobacterium tuberculosis
Tissue Culture Media: Chicken Embryo
Source:
Fertilized Chicken Egg
The transfer of microorganisms from one culture media to another.
Transplant or Subculture
HEA Agar
- Selective for:
Gram negative enteric bacilli
Antibiotic Inhibitors for Gram Negative Bacteria
- Colistin
- Nalidixic Acid
- Trimethoprim
Most pathogenic bacteria can grow in an environment with a pH of ________.
7.0 - 7.5
The generation time of bacteria in culture media is __________.
20 minutes to 24 hours
Hyperthermophilic or Exteremely Thermophilic
80-110 C
HEA Agar
- Original Color:
Dark Green
Dye Inhibitors for Gram Negative Bacteria
- Basic Fuchsin
- Thionine
Martin-Lewis Agar
Fungal Inhibitor:
Anisomycin
MSA Agar
- Inhibitors:
High Concentration of Salt (7.5%)
A group of bacteria that utilizes organic compounds in the culture media as a source of nutrition.
Heterotrophs
It is used for the determination of bacterial pathogen’s ability to ferment glucose, sucrose, and lactose.
Triple Sugar Iron Agar (TSI)
SSA Agar
- Original Color:
Light Orange
MAC Agar
- Selective for:
Gram negative enteric bacilli
A culture medium that contains 0.5 - 1% agar.
Semi-solid
It has a medium size, dark violet colonies with greenish metallic sheen.
Escherichia
SSA Agar
- pH Indicators:
Neutral Red
The act of introducing the microorganism to the culture media.
Inoculate/Plant/Cultivate
New York City Agar
Fungal Inhibitor:
Amphotericin B
These are live cells harvested from organs of humans and animals that supports the growth of obligate intracellular organisms that cannot grow in artificially prepared culture media.
Tissue Culture Media
Tissue Culture Media: Help-2 Cell Line
Source:
Human epithelial cells of larynx carcinoma
LJ Medium
- Incubation without light = pigment
Scotochromogen
Example of Alkaliphilic Bacteria
Gardnerella vaginalis
SSA Agar
Lactose fermenters will appear _________.
Dark Pink
It is composed of known and exact amounts of pure chemical substances.
Synthetic or Chemically Defined
An enrichment broth for anaerobic bacteria.
Thioglycolate Broth
LJ Medium
- Inhibitors:
Malachite Green
It refers to the time it takes for one cell to divide into two cells.
Generation or Doubling Time
MAC AGAR
Non-lactose fermenters will appear _________.
Colorless
It contains basic nutritional requirements to support the growth of non-fastidious microorganisms.
Gen.Purpose/Primary/Basic/Basal/Supportive/General Isolation Culture Media
It is a liquid enrichment recommended for the production of Streptococci haemolysin and the cultivation of it.
Todd-Hewitt Broth
Nitrogen Sources
- Free Nitrogen from the air
- Nitrogenous Compounds in the Culture Media
Most common bottle used in media.
bacT alert
It contains indicators which changes in color as a result of a product produced by a chemical reaction in the media.
Differential Culture Media
SSA Agar
- H2S Indicator:
Ferric ammonium citrate
It can tolerate up to 10% of salt concentration.
Vibrio spp.
In LJ medium, before culturing, the sample must undergo the following steps ___________.
- Decongested/Digested
- Decontaminated
Chemical Inhibitors for Gram Positive Bacteria
- Bismuth sulfite
- Bile Salts (sodium deoxycholate)
- Thiosulfate
- Citrate
TCBS Agar
Sucrose fermenters will appear _________.
Yellow
_________ is needed for the cell wall.
Phospholipid
TCBS Agar:
- Original Color:
Light Green or Olive Green
These are bacteria that absolutely requires oxygen to grow, live, and survive.
Strict or Obligate Aerobes
Carbon Dioxide Sources
- Carbon Dioxide from the air
- Organic Compounds in the Culture Media
A primary media to support or favor the selective growth of pathogens in a specimen, where the number of normal flora outnumber the pathogens.
Enrichment Broth
__________ is needed for protein synthesis.
Sulfur
HEA Agar
Lactose fermenters will appear _________.
Orange
New York City Agar
Gram Negative Inhibitor:
Colistin
BAP contains which factor/s?
X only
Selective medium composition for Neisseria spp.
Chocolate agar base with antibiotics
These are bacteria that can survive high salt concentrations.
Halophiles
MAC Agar
- Original Color:
Pink
Most pathogenic bacteria grow at temperature between __________.
35-37 C
It is used for the production of hydrogen sulfide gas, indole production, and motility.
Sulfide Indole Motility Medium (SIM)
Thayer Martin Agar
Fungal Inhibitor:
Nystatin
It is used to selectively favor the growth of Salmonella and Shigella while inhibiting all other normal intestinal flora.
Gram Negative Broth (GN)
Which Stage of Bacterial Growth?
- The phase where bacteria is most metabolically active.
Logarithmic Phase
RLF can produce a result within __________.
18 to 24 hours
EMG Agar
- Differential Indicators:
Eosin and Methylene Blue
A group of bacteria that uses Carbon Dioxide from the air as a source of nutrition.
Autotrophs
Why is agar used as a solidifying agent?
- Not easily degraded by bacteria
- Can melt and re-solidify easily
EMG Agar
- Inhibitors:
Eosin and Methylene Blue
Modified Thayer Martin Agar
Fungal Inhibitor:
Nystatin
LJ Medium
- Incubation with light = pigment
Photochromogen
It is used for the detection of bacterial pathogen that metabolizes glucose using the Mixed Acid Pathway.
Methyl Red (MR)
These are bacteria that are capable of getting an electron from inorganic molecules.
Lithotrophs
MSA Agar
Non-mannitol fermenters will appear _________.
Dark Pink or Red
SSA Agar
- Inhibitors:
Brilliant Green, Bile Salts, and Citrate
TCBS Agar
Non-sucrose fermenters will appear _________.
Blue-Green
These are bacteria that grow, live, and survive in the absence of oxygen.
Anaerobes
BSA Agar
- Selective for:
Salmonella spp. (specifically Typhi)
BSA Agar
- Inhibitors:
Bismuth sulfite
BGA Agar
- Inhibitors:
Brilliant Green
Rapid Lactose Fermenters
- Escherichia
- Klebsiella
- Enterobacter
HEA Agar
- Sulfur Source:
Sodium Thiosulfate
Decontaminated means _________.
to eliminate normal flora that contaminates the sample using NaOH
TCBS Indicator:
- pH Indicator
Bromothymol Blue
It has a medium size, dark violet colonies with or without dark center.
Enterobacter
These are bacteria that prefers small concentration of oxygen environment; approximately 2-10%.
Microaerophilic Bacteria
The standard size of plated media used in AST.
150
It is used to detect bacterial pathogen that hydrolyze urea substrate.
Stuart’s Urea Broth or Christensen Urea Agar
To grow or to cultivate microorganisms
Culture (verb)
It is needed in order for the cell to perform its metabolic or cellular functions.
Energy
SSA Agar
Non-lactose fermenters will appear _________.
Colorless
Example of an Osmophilic Bacteria
Archaebacteria spp.
A tube commonly used in media.
Wasserman tubes
Martin-Lewis Agar
Gram Negative Inhibitor:
Colistin
It is used for the detection of bacterial pathogen that can utilize citrate as a sole source of carbon.
Simmon Citrate Agar (SCA)
SSA Agar
- Shigella Result:
Colorless, w/o H2S and w/o blackening
These are needed as co-factors in various metabolic processes of the bacteria.
Mineral Elements
Other sources besides CNE
- Phosphates
- Phospholipids
- Sulfur
LJ Medium
- Selective for:
Mycobacterium spp.
Thermophilic
50-60 C
It is composed of complex materials that are rich in vitamins and nutrients that are not usually represented by a chemical formula.
Non-Synthetic or Non-Chemically Defined
Decongested or Digested means _________.
to dissolve the thick mucin or mucus using N-acetyl L-cysteine (NALC)
These are bacteria that grow, live, and survive in the presence of oxygen.
Aerobes
Classification of Culture Media According to Composition
- Synthetic or Chemically Defined
- Non-Synthetic or Non-Chemically Defined
- Tissue Culture Media
HEA Agar
- H2S Indicator:
Ferric ammonium citrate
SSA Agar
- Selective for:
Salmonella spp. and Shigella spp.
An example of a heterotroph is _________.
Most normal flora in our body
Sources of Energy
- Light
- Chemical Energy
LJ Medium
- Original Color:
Light Green
It is used to detect lysine decarboxylation, ornithine decarboxylation, and arginine dehydrolysis.
Moeller’s Broth
Modified Thayer Martin Agar
Gram Positive Inhibitor:
Vancomycin
It favors the growth of the organism of interest using inhibitors added in the culture media.
Selective Culture Media
Examples of Enrichment Borth
- Alkaline Peptone Water (APW)
- Selenite F. Broth
- Thioglycolate Broth
- GN Broth
- Todd-Hewitt Broth
Tissue Culture Media: Mc Coy Cell Line
Source:
Mouse Cell Line
Late Lactose Fermenters
- Hafnia
- Serratia
- Citrobacter
- Salmonella arizonae
- Shigella sonnei
- Yersinia enterocolitica
Thayer Martin Agar
Gram Negative Inhibitor:
Colistin
Bacteria can tolerate salt concentration _______.
Below 6%
EMG Agar
- Original Color:
Dark Violet
BGA Agar
- CHO Incorporated:
Lactose
Sternothermophilic
Narrow range of temperature
Examples of Enriched Culture Media
Chocolate Agar Plate, Blood Agar Plate
LJ Medium
- No pigment production
Non-photochromogen
It is usually a container in glass tubes with different volume capacity.
Tubed
These are bacteria that are capable of getting an electron from organic molecules.
Organotrophs
It contains the basic nutritional requirements with additives, enriching substances, or supplements to support the growth of fastidious microorganisms.
Enriched Culture Media
It is used to selectively favor the growth of Salmonella while inhibiting all other normal intestinal flora.
Selenite F. Broth
Classification of Culture Media According to Physical State or Consistency
- Liquid
- Semi-solid
- Solid
Which Stage of Bacterial Growth?
- Essential nutrients are depleted.
Stationary Phase
MAC Agar
- Inhibitors:
Crystal violet, bile salts, and citrate
LLF can produce a result within ___________.
36 to 72 hours
Which Stage of Bacterial Growth?
- The rate of cell division equates the rate of cell death.
Stationary Phase
________ is needed for nucleic acids.
Phosphates
Bacteria that can tolerate high osmotic pressure
Osmophilic
Mesophilic
20-40 C
Thayer Martin Agar
Gram Positive Inhibitor:
Vancomycin
Psychrophilic or Cryophilic
0-20 C
Antibiotic Inhibitors for Gram Positive Bacteria
Vancomycin
Classification of Culture Media According to Manner of Dispensing or Formation
- Plated
- Tubed
- Bottled
New York City Agar
Gram Positive Inhibitor:
Vancomycin
It is needed for the synthesis of proteins and nucleic acid.
Nitrogen
The X Factor is also known as ________.
Hemin/Hematin
These are bacteria that can grow, live, and survive in small concentrations of oxygen (w/wo).
Facultative Anaerobes
GC-LECT Agar
Gram Negative Inhibitor:
Colistin
Examples of Transport Culture Media
- Stuart’s Transport Medium
- Cary-Blair
- Amie’s
- Transgrow
- JEMBEC
Examples of Capnophilic Bacteria
- Neisseria spp.
- Haemophilus spp.
- Aggregatibacter spp.
- Cardiobacterium spp.
- Eikenella
- Kingella
Examples of Aerotolerant Anaerobes
- Clostridium perfringens
- Bacteroides fragilis
- Propionibacterium
- Lactobacillus
It is typically used for decongestion/digestion.
N-acetyl L-cysteine
Which Stage of Bacterial Growth?
- Toxic products accumulate
Stationary Phase
These are bacteria that do not require oxygen but may tolerate or withstand limited oxygen exposure.
Aerotolerant Anaerobes
A culture medium that contains 1.5 - 3% agar.
Solid
MAC AGAR
Lactose fermenters will appear _________.
Dark Pink
Which Stage of Bacterial Growth?
- The bacteria is still adjusting to the environment.
Lag Phase
It is used for the detection of bacterial pathogen that metabolizes glucose using the Butylene Glycol Pathway.
Voges-Proskauer (VP)
It refers to the increase in the number of bacteria rather than in size.
Bacterial Growth
Examples of Strict/Obligate Anaerobes
- Clostridium noyvi
- Bacteroides spp.
- Fusobacterium spp.
- Peptostreptococcus spp.
- Porphyromonas spp.
These are bacteria that requires 5-10% of CO2 to grow.
Capnophilic Bacteria
MSA Agar
- Indicator:
Phenol Red
A primary isolation culture media which maintains the viability of bacteria without allowing rapid multiplication.
Transport Culture Media
Bacteria is made up of _____% water.
70
Chemical Inhibitors for Gram Negative Bacteria
- Potassium Tellurite
- Sodium Azide
- Phenlyethyl Alcohol
Energy is in the form of __________.
Adenosine Triphosphate (ATP)
Describe the colony appearance of Salmonella spp. in BSA agar
Black colonies with silver metallic sheen
The V Factor is also known as ________.
Nicotinamide Adenine Dinucleotide (NAD)
Example of Acidophilic Bacteria
Lactobacillus acidophilus
It is used to selectively favor the growth of Vibrio while inhibiting all other normal intestinal flora due to its high pH.
Alkaline Peptone Water (APW)
Which Stage of Bacterial Growth?
- The phase where bacteria is most susceptible to the action of antimicrobial agents.
Logarithmic Phase
It is the degradation product of hemoglobin.
Hemin/Hematin (X Factor)
Tissue Culture Media: Vero Cell Line
Source:
Kidney Cells of African Green Monkey
BSA Agar
- CHO Incorporated:
Glucose
MSA Agar
- Selective for:
Staphylococcus spp.
TCBS Agar
- CHO incorporated
Sucrose
It has a large size, mucoid, dark violet colonies with or without dark center.
Klebsiella
Culture media contained in a glass bottle that is usually used for blood culture.
Bottled
Give examples of halophilic bacteria
- Staphylococcus spp.
- Enterococcus spp.
- Vibrio spp. (except cholera and mimicus)
- Bacillus spp.
A culture medium that does not have a solidifying agent.
Liquid
Nitrogen makes up _______% of the dry weight of a bacterium.
14
A fast-growing bacteria is ___________.
Escherichia coli
What are the 2 major bacterial growth requirements?
Nutritional and Environmental
These are bacteria that absolutely do not require oxygen to grow, live, and survive.
Strict or Obligate Anaerobes
Modified Thayer Martin Agar
Proteus Inhibitor:
Trimethoprim
It is needed for the synthesis of cellular components.
Carbon
EMB AGAR
Lactose fermenters will appear _________.
Dark Violet
HEA Agar
- pH Indicators:
Bromothymol Blue (BTB)
Bacterial metabolism without the use of oxygen.
Fermentation
Electron Sources
- Inorganic Molecules
- Organic Molecules
- NADH
- FADH2
Examples of Microaerophilic Bacteria
- Campylobacter spp.
- Helicobacter spp.
- Arcobacter spp.
- Streptococcus spp. (some)
Which Stage of Bacterial Growth?
- Bacterial cell division decrease, while cell death becomes more accelerated.
Death Phase
Which Stage of Bacterial Growth?
- No cell division occurs, but there are already cellular activities.
Lag Phase
Carbon represents ______% of the dry weight of a bacterium.
50
HEA Agar
Non-lactose fermenters will appear _________.
Blue/Green
It is usually grown in MSA that has 7% of NaCl
Staphylococcus spp.
BGA Agar
- Selective for:
Salmonella spp. (except Typhi)
TCBS Agar
- Selective for:
Vibrio spp.
An agar plate that has lysed RBC.
Chocolate Agar Plate (CAP)
Describe the colony appearance of Salmonella spp. in BGA agar
White colonies surrounded by brilliant red medium
Commonly, the _________ bacteria are extremely thermophilic.
spore-forming
