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Culturing Microorganisms > Culturing Bacteria
Culturing Bacteria
• Culture Media
• Complex and Synthetic Media
• Selective and Differential Media
• Aseptic Technique, Dilution, Streaking, and Spread Plates
• Special Culture Techniques
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Culturing Microorganisms > Culturing Bacteria
Culture Media
• Culture media contains the nutrients needed to sustain a microbe.
• Culture media can vary in many ingredients allowing the media to select for or
against microbes.
• Glucose or glycerol are often used as carbon sources, and ammonium salts or
nitrates as inorganic nitrogen sources in culture media.
Microbial pathogen growing on blood-agar plate
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Culturing Microorganisms > Culturing Bacteria
Complex and Synthetic Media
• Defined media is made from constituents that are completely understood.
• Undefined media has some part of which is not entirely defined.
• The presence of extracts from animals or other microbes makes a media
undefined as the entire chemical composition of extracts are not completely
known.
Undefined Media
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Culturing Microorganisms > Culturing Bacteria
Selective and Differential Media
• Selective media generally selects for the growth of a desired organism, stopping
the growth of or altogether killing non-desired organisms.
• Differential media takes advantage of biochemical properties of target organisms,
often leading to a visible change when growth of target organisms are present.
• Differential media, unlike selective media, does not kill organisms. It indicates if a
target organism is present.
Non-selective versus selective media.
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Culturing Microorganisms > Culturing Bacteria
Aseptic Technique, Dilution, Streaking, and Spread Plates
• Aseptic technique is basically the mindset of keeping things free of contamination,
as the world we live in has so many microbes that can interfere with experiments.
• Colony streaking leads to to the isolation of individual colonies, which are a group
of microbes that came from one single progenitor mircrobe.
• Spread plates allow for the even spreading of bacteria onto a petri dish; allowing
for the isolation of individual colonies, for counting or further experiments.
Serial Dilution
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Culturing Microorganisms > Culturing Bacteria
Special Culture Techniques
• Microbes, often those that we know little about, have to be cultured with undefined
media or growth conditions.
• The use of animals to culture animals is sometimes necessary as no simple
media can be used, this presents technical and ethical issues.
• As human pathogens are often studied by microbiologists, special safety
conditions know as biosafety levels are used to keep researches free of infection
from the pathogens they study.
Candle jar
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Appendix
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Culturing Microorganisms
Key terms
• allele One of a number of alternative forms of the same gene occupying a given position on a chromosome.
• bunsen burner A small laboratory gas burner whose air supply may be controlled with an adjustable hole.
• colony A bacterial colony is defined as a visible cluster of bacteria growing on the surface of or within a solid medium,
presumably cultured from a single cell.
• culture The process of growing a bacterial or other biological entity in an artificial medium.
• gene A unit of heredity; a segment of DNA or RNA that is transmitted from one generation to the next. It carries genetic
information such as the sequence of amino acids for a protein.
• Lyme disease Infection by a bacterium of the genus Borrelia which is transmitted by ticks. Symptoms include a rash followed by
fever, joint pain, and headaches.
• lysogeny broth Lysogeny broth (LB) is a nutritionally-rich medium; primarily used for the growth of bacteria.
• recombinant This term refers to something formed by combining existing elements in a new combination. Thus, the phrase
recombinant DNA refers to an organism created in the lab by adding DNA from another species.
• recombinant This term refers to something formed by combining existing elements in a new combination. Thus, the phrase
recombinant DNA refers to an organism created in the lab by adding DNA from another species.
• serum The clear yellowish fluid obtained upon separating whole blood into its solid and liquid components after it has been
allowed to clot. Also called blood serum.
• yellow fever An acute febrile illness of tropical regions, caused by a flavivirus and spread by mosquitoes, characterized by
jaundice, black vomit, and the absence of urination.
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Culturing Microorganisms
Microbial pathogen growing on blood-agar plate
Red blood cells are used to make an agar plate. Different pathogens that can use red blood cells to grow are shown on these plates. On the left is
staphylococcus and the right streptococcus.
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Wikimedia. "Agarplate redbloodcells edit." Public domain http://commons.wikimedia.org/wiki/File%253AAgarplate_redbloodcells_edit.jpg View on Boundless.com
Culturing Microorganisms
Serial Dilution
Example of Serial dilution of bacteria in five steps. The diluted bacteria were then spread plated.By Leberechtc (Own work) [GFDL
(http://www.gnu.org/copyleft/fdl.html) or CC-BY-3.0 (http://creativecommons.org/licenses/by/3.0)], via Wikimedia Commons
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Wikimedia. "Verd%C3%BCnnungsreihe%20mit%20Ausplattieren." CC BY
http://commons.wikimedia.org/wiki/File%25253AVerd%2525C3%2525BCnnungsreihe_mit_Ausplattieren.svg View on Boundless.com
Culturing Microorganisms
Chlamydias bacteria group
Light microscope view of cells infected with chlamydiae as shown by the brown inclusion bodies.
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Wikipedia. "ChlamydiaTrachomatisEinschlusskörperchen." CC BY 3.0
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Culturing Microorganisms
Undefined Media
Luria Broth as shown here is made with yeast extract, as yeast extract is not completely chemically defined Luria Broth is therefore an undefined
media.By Lilly_M [GFDL (http://www.gnu.org/copyleft/fdl.html) or CC-BY-SA-3.0-2.5-2.0-1.0 (http://creativecommons.org/licenses/by-sa/3.0)], via
Wikimedia Commons
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Wikimedia. CC BY-SA http://upload.wikimedia.org/wikipedia/commons/c/c1/LB_luquid_medium_bottle-01.jpg View on Boundless.com
Culturing Microorganisms
Candle jar
A candle is lit in a jar with a culture plate. The lid is put on, as the burns it increases the carbon dioxide levels in the jar.
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Wikimedia. "Anaerobic chamber." CC BY-SA http://commons.wikimedia.org/wiki/File%253AAnaerobic_chamber.JPG View on Boundless.com
Culturing Microorganisms
Positive pressure suit
A scientist puts on a positive pressure suit, something needed to work with the most dangerous human pathogens in a biosafety level 4 laboratory.
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Culturing Microorganisms
Non-selective versus selective media.
The non-selective media on the right allows of the growth of several different bactarial species and is overgrown with bacteria (whitish lines). While the
plate on the right selectively only allows the bacteria Neisseria gonorrhoeae, to grow (white dots).
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Wikimedia. "Neisseria gonorrhoeae 01." Public domain http://commons.wikimedia.org/wiki/File:Neisseria_gonorrhoeae_01.png View on Boundless.com
Culturing Microorganisms
Streak plate
Four streak plates. Successful streaks lead to individual colonies of microbes.
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Culturing Microorganisms
Which of the following are types of media?
A) differential
B) defined
C) All of the above answers
D) minimal
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Culturing Microorganisms
Which of the following are types of media?
A) differential
B) defined
C) All of the above answers
D) minimal
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Boundless - LO. "Boundless." CC BY-SA 3.0 http://www.boundless.com/
Culturing Microorganisms
You have a media that you know the buffer, salts and everything
consituent in it. You add an extract from sheep muscle to the
media. What type of media is it?
A) Defined
B) Synthetic
C) Undefined
D) Selective
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Culturing Microorganisms
You have a media that you know the buffer, salts and everything
consituent in it. You add an extract from sheep muscle to the
media. What type of media is it?
A) Defined
B) Synthetic
C) Undefined
D) Selective
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Boundless - LO. "Boundless." CC BY-SA 3.0 http://www.boundless.com/
Culturing Microorganisms
You find a new yeast cell that cannot produce its own amino acid,
specifically alanine. Therefore, if you were to plate out different
yeast cells on media lacking alanine, your yeast strain would not
grow. What type of media would this be?
A) Differential
B) Selective
C) YM
D) X-gal
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Culturing Microorganisms
You find a new yeast cell that cannot produce its own amino acid,
specifically alanine. Therefore, if you were to plate out different
yeast cells on media lacking alanine, your yeast strain would not
grow. What type of media would this be?
A) Differential
B) Selective
C) YM
D) X-gal
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Boundless - LO. "Boundless." CC BY-SA 3.0 http://www.boundless.com/
Culturing Microorganisms
What is the common goal of streaking or spreading bacteria on a
plate?
A) Counting cells
B) Checking for contamination
C) Isolation of individual colonies
D) Serial dilution
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Culturing Microorganisms
What is the common goal of streaking or spreading bacteria on a
plate?
A) Counting cells
B) Checking for contamination
C) Isolation of individual colonies
D) Serial dilution
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Boundless - LO. "Boundless." CC BY-SA 3.0 http://www.boundless.com/
Culturing Microorganisms
You have discovered a new type of capnophile; where would you
culture it?
A) armadillo
B) biohood
C) McCoy culture cells
D) candlejar
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Culturing Microorganisms
You have discovered a new type of capnophile; where would you
culture it?
A) armadillo
B) biohood
C) McCoy culture cells
D) candlejar
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Culturing Microorganisms
Attribution
• Wiktionary. "serum." CC BY-SA 3.0 http://en.wiktionary.org/wiki/serum
• Wiktionary. "recombinant." CC BY-SA 3.0 http://en.wiktionary.org/wiki/recombinant
• Wikipedia. "Chemically defined medium." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Chemically_defined_medium
• Wikipedia. "Growth medium." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Growth_medium
• Wikipedia. "Growth medium." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Growth_medium
• Wikipedia. "Culture media." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Culture_media
• Wikipedia. "Culture media." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Culture_media
• Wikipedia. "Culture media." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Culture_media
• Wikipedia. "Culture media." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Culture_media
• Wikipedia. "Culture media." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Culture_media
• Wikipedia. "Culture media." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Culture_media
• Wikipedia. "Culture media." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Culture_media
• Wikipedia. "lysogeny broth." CC BY-SA 3.0 http://en.wikipedia.org/wiki/lysogeny%20broth
• Wiktionary. "culture." CC BY-SA 3.0 http://en.wiktionary.org/wiki/culture
• Wikipedia. "Growth medium." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Growth_medium
• Wiktionary. "allele." CC BY-SA 3.0 http://en.wiktionary.org/wiki/allele
• Wiktionary. "gene." CC BY-SA 3.0 http://en.wiktionary.org/wiki/gene
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Culturing Microorganisms
• Wiktionary. "recombinant." CC BY-SA 3.0 http://en.wiktionary.org/wiki/recombinant
• Wikipedia. "Growth medium." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Growth_medium
• Wikia. "Sterile Technique - Laboratory Wiki." CC BY-SA http://lab.wikia.com/wiki/Sterile_Technique
• Wikipedia. "Serial dilution." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Serial_dilution
• Wikipedia. "Streaking (microbiology)." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Streaking_(microbiology)
• Wikipedia. "Serial dilution." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Serial_dilution
• Wiktionary. "bunsen burner." CC BY-SA 3.0 http://en.wiktionary.org/wiki/bunsen+burner
• Wikipedia. "colony." CC BY-SA 3.0 http://en.wikipedia.org/wiki/colony
• Wikipedia. "Biosafety level." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Biosafety_level
• Wikipedia. "Biosafety level." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Biosafety_level
• Wikipedia. "Biosafety level." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Biosafety_level
• Wikipedia. "Armadillo." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Armadillo
• Wikipedia. "Armadillo." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Armadillo
• Wikipedia. "Chlamydia infection." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Chlamydia_infection
• Wikipedia. "Growth medium." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Growth_medium
• Wikipedia. "Microaerophile." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Microaerophile
• Wikipedia. "Growth medium." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Growth_medium
• Wikipedia. "Biosafety level." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Biosafety_level
• Wikipedia. "yellow fever." CC BY-SA 3.0 http://en.wikipedia.org/wiki/yellow%20fever
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Culturing Microorganisms
• Wikimedia. "ChlamydiaTrachomatisEinschlusskörperchen." CC BY-SA 3.0
http://commons.wikimedia.org/wiki/File:ChlamydiaTrachomatisEinschlussk%25C3%25B6rperchen.jpg
• Wikipedia. "Biosafety level." CC BY-SA 3.0 http://en.wikipedia.org/wiki/Biosafety_level
• Wiktionary. "Lyme disease." CC BY-SA 3.0 http://en.wiktionary.org/wiki/Lyme+disease
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