1. Bacterial Biotechnology

2. The Structure of Microbes
Prokaryotes
Archaebacteria
Includes halophiles, thermophiles, “extremophiles”
Eubacteria
On skin, pathogens, soil, water
Generally smaller than Eukaryotes (1-5μm vs μm)
What are some other characteristics of prokaryotes? (cell wall (gram stain), no nucleus, binary fission, 20 min growth rate…)
Do you know how to isolate single colonies?

3. Microorganisms as Tools
Microbial Enzymes
Taq (DNA polymerase), cellulases, proteases, amylases
Bacterial Transformation
The ability of bacteria to take in DNA from their surrounding environment
Cells must be made competent (to take up DNA)

4. Figure 5.3

5. Electroporation
A mixture of bacteria and plasmid are briefly electrically shocked

6. Figure 5.4

7. Cloning and Expression Techniques
Fusion Proteins
Use recombinant DNA methods to insert the gene for a protein of interest into a plasmid containing a gene for a well-known protein that serves as a “tag”
The tag allows for isolation and purification
Ex. GFP – Green Fluorescent Protein
Used to follow certain molecules through cell processes

8. Figure 5.5

9. Microbial Proteins as Reporters
Examples: the lux gene which produces luciferase
Used to develop a fluorescent bioassay to test for TB (the lux gene is in a virus that only infects M. tuberculosis). If the bacteria is present, the virus infects the cells and the bacterial cells glow!

10. Using Microbes for a Variety of Everyday Applications
Food Products
Rennin used to make curds (solid) and whey in production of cheese
Recombinant rennin is known as chymosin (first recombinant food product approved by FDA)

11. Therapeutic proteins Recombinant insulin in bacteria
What is Type I diabetes (insulin-dependent diabetes mellitus)
Inadequate production of insulin by beta cells in the pancreas

12. Figure 5.9

13. Field Applications of Recombinant Microorganisms
Ice-minus bacteria (remove ice protein producing genes from P. syringae)
P. fluorescens containing the gene that codes for the bacterial toxin from Bacillus thuringiensis (kills insects) Bt toxin!

14. Using Microbes Against Other Microbes
Antibiotics
Penicillin was the first
Act in a few key ways
Prevent replication
Kill directly
Damage cell wall or prevent its synthesis
How do antibiotic resistant strains arise?
How can studying bacterial pathogens lead to new drugs?

15. Figure 5.10

16. Microbial Genomes
Microbial Genome Program (MGP) –the goal is to sequence the entire genomes of microorganisms that have potential applications in environmental, biology, research, industry, and health
Sequencing Strategies

17. Figure 5.15

18. Microbial Diagnostics
Using Molecular Techniques to Identify Bacteria
RFLP - restriction fragment length polymorphism
PCR – Polymerase Chain Reaction
PulseNet
(Contaminated food) - a network run by the Centers for Disease Control and Prevention (CDC) which brings together public health and food regulatory agency laboratories around the United States

19. Figure 5.16

20. Combating Bioterrorism
The use of biological materials as weapons to harm humans or animals and plants we depend on for food
Examples in History
Throwing plague infected dead bodies over the walls of their enemies
Using Biotech Against Bioweapons
Postal service x-raying packages
Antibody tests in the field
PCR tests in the field

21. An intro to bacteria, infectious diseases, and antibiotic resistance
Image: Wikipedia (
RISE OF THE SUPERBUGS!
An intro to bacteria, infectious diseases, and antibiotic resistance

22. An intro to bacteria, infectious diseases, and antibiotic resistance
Image: Wikipedia (
RISE OF THE SUPER BUGS!
An intro to bacteria, infectious diseases, and antibiotic resistance

23. How abundant are bacteria?
Bacterial Abundance
Total bacteria on Earth
5 x 1030
Number of stars in the universe
7 x 1022
Age of the universe in seconds
4.4 x 1017
Bacteria in the human gut
1 x 1014
Global gross product ($/year)
7 x 1013
Cells in the human body
1 x 1013
Texts sent in 2009
1.5 x 1012
People on Earth
6.9 x 109
10X
“Abundance” = how many…
Notice that there are 10 times more bacterial cells in your gut alone than there are human cells in your entire body!
Do you think you are sick because they are there, or that you are healthy because they are there?

24. How many types of bacteria are there?
Bacterial Distribution
Insect Species
1-10 million
Bacterial species in the soil
4 million
Bacterial species in the air
Bacterial species in the ocean
2 million
Bird Species
10,000
Bacterial species in the human mouth
600
Bacterial Species in the human gut
500
Pathogenic Species 55
Type = species
Note that there are really few harmful bacteria compared to all others. MOST BACTERIA ARE HARMLESS OR HELPFUL!

25. Infectious Diseases Pathogen: a biological agent that causes disease
Can be a virus, bacteria, fungi, protozoa, or parasite
Infectious diseases are communicable or transmissible from one person to another
Pathogens are microorganisms that make you sick/ill.

26. Protist: Malaria (Plasmodium)
Virus: Influenza (H1N1)
Protist: Malaria (Plasmodium)
Fungi: Plant pathogen (Rigidoporus laetus)
H1N1 Virus:
Marlaria:
Fungi:

27. Staph Infection: Staphylococcus aureus
Mycobacterium tuberculosis
Salmonella typhimurium
Tuberculosis:
Salmonella:
Staph:

28. Antibiotics Compounds that kill bacteria or inhibit their growth
Naturally produced by microorganisms to kill others
Now many are made synthetically
Antibiotics are used to treat bacterial infections.
Antimicrobials are a broader class of chemical/compounds that kill bacteria, fungi, and protists.
Antibiotics can kill a bacteria in several different ways. Some target DNA or protein synthesis, others interrupt the cell membrane.
Prontosil, the first commercially available antibacterial antibiotic was developed by a research team led by Gerhard Domagk (who received the 1939 Nobel Prize for Medicine for his efforts) at the Bayer Laboratories of the IG Farben conglomerate in Germany.
The development of penicillin led to renewed interest in the search for antibiotic compounds with similar capabilities.[17] Because of their discovery of penicillin Ernst Chain, Howard Florey and Alexander Fleming shared the 1945 Nobel Prize in Medicine.

29. Why is resistance bad? Because you can’t treat the disease!
The patient will remain sick 

30. How do you get resistance?
Usually from mutation in DNA
Genetic mutations can be passed to offspring…
Leads to evolution!
Evolution is the change in the inherited traits of a population of organisms through successive generations. (Changes in a population over time).

31. Genetic Mutation
NATURAL SELECTION!

32. Susceptible
+ Antibiotic
Antibiotic Resistant

33. What is MRSA?

34. MRSA MRSA stands for Methicillin-Resistant Staphylococcus aureus
MRSA are Staph aureus bacteria that have become resistant to this antibiotic.
Found in the nose, and sometimes on the skin, but also can grow in wounds and other sites of the body
MRSA can no longer be killed by this antibiotic

35. Recurrent skin diseases or open wounds
In general, healthy people are at low risk of getting sick with MRSA. Some risk factors include:
Recurrent skin diseases or open wounds
Long-term illness or long-term dialysis patient
Illicit injecting drug use
Surgery
Those with weakened immune systems from sickness or chronic disease are the most vulnerable.

36. Contact with other persons with MRSA infection
Been a patient in the hospital or other health care facility within the past year
Contact with other persons with MRSA infection
Recent antibiotic use
Live in crowded settings
MRSA infections have been reported among persons in prisons, players of close-contact sports, men who have sex with other men, and other populations.

37. HOW ARE THESE GERMS SPREAD?
MRSA is transmitted primarily by contact with a person who has an infection or is colonized with the bacteria.
The germ can be spread by direct person-to-person contact with an infected person, or by contact with objects or surfaces contaminated by MRSA.
Staph can also come off infected skin onto shared objects and surfaces and get onto the skin of the person who uses the object or surface next.
Examples of shared objects that might spread staph include personal hygiene objects (i.e. towels, soap, wound dressings, bandages, etc.), sheets, clothes, benches in saunas or hot tubs, and athletic equipment.
In other words, anything that could have touched the skin of a staph-infected person can carry the bacteria to the skin of another person.

38. And now, let’s look at what MRSA really looks like…

39. MRSA/Staph is often misdiagnosed as spider or insect bites.
• MRSA/Staph spreads by infected skin to healthy skin contact as well as infected objects
to healthy skin. It can enter healthy, clean, undamaged skin through such contact.
• Washing your hands with soap and warm water can prevent the spread of MRSA/Staph.
• MRSA/Staph lives on skin and survives on objects, such as towels and exercise equipment for 24 hours or longer.
• If you think you may have MRSA/Staph, consult your doctor or healthcare provider.
• For all skin infections, dispose of bandages properly and wash your hands frequently to avoid spreading germs to others.

40. This I do believe is self-explanatory
This I do believe is self-explanatory! The single most important thing that you can do to stop the spread of any germs is to wash your hands

41. NDM-1 New Delhi Metallo-beta-lactamase-1 (NDM-1)
an enzyme that makes bacteria resistant to a broad range of beta-lactam antibiotics
first detected in a Klebsiella pneumoniae isolate from a Swedish patient of Indian origin in 2008
Later detected in India, Pakistan, the United Kingdom, the United States, Canada, and Japan
Evolution is the change in the inherited traits of a population of organisms through successive generations. (Changes in a population over time).

42. Usually found in: Escherichia coli and Klebsiella pneumoniae,
but the gene for NDM-1 can spread from one strain of bacteria to another by horizontal gene transfer!
Evolution is the change in the inherited traits of a population of organisms through successive generations. (Changes in a population over time).

43. KPCs Klebsiella pneumoniae carbapenemases (KPCs)
2001, North Carolina - first KPC-producing K. pneumoniae isolate reported
mechanism of resistance for a range of Gram-negative bacteria
often not detected by routine susceptibility screening
present serious treatment challenges, due to limited antibiotic options
Evolution is the change in the inherited traits of a population of organisms through successive generations. (Changes in a population over time).

44. Usually found in: Klebsiella pneumoniae,
no longer limited to K. pneumoniae
Enterobacteriaceae-producing KPCs have also been reported in Brazil, China, Colombia, Norway, United Kingdom, India, Sweden, and more recently, Italy and Finland
Evolution is the change in the inherited traits of a population of organisms through successive generations. (Changes in a population over time).

45. Acknowledgements
The Infectious Disease Control Unit of the Department of State Health Services thanks
Ginger Shields, RN
Emergency Preparedness Specialist
Texarkana-Bowie County Health Department
for the creation and use of this presentation.