1. The Genetics of Antibiotic Resistance Research Theme: Infectious Diseases Jason Kuehner March 5, 2007

2. http://scientificteaching.wisc.edu/video

3. What’s wrong with this statement? Take a few minutes to individually complete this worksheet and then compare your answers with your group

4. What’s wrong with this statement? Take a few minutes to individually complete this worksheet and then compare your answers with your group Consider these misconceptions throughout class today and we’ll follow-up on them at the end

5. The Problem

6. You run a public health clinic in Racine, Wisconsin

7. The Problem You run a public health clinic in Racine, Wisconsin A county commissioner overseeing your clinic is an epidemiologist and wants to know how you plan to address the emergence of ciprofloxacin resistance in Neisseria gonorrhoeae

8. The Problem You run a public health clinic in Racine, Wisconsin A county commissioner overseeing your clinic is an epidemiologist and wants to know how you plan to address the emergence of ciprofloxacin resistance in Neisseria gonorrhoeae Medical professional that deals with the incidence, distribution, and possible control of diseases

9. The Problem You run a public health clinic in Racine, Wisconsin A county commissioner overseeing your clinic is an epidemiologist and wants to know how you plan to address the emergence of ciprofloxacin resistance in Neisseria gonorrhoeae Oral antibiotic approved for treatment of many common bacterial infections

10. The Problem You run a public health clinic in Racine, Wisconsin A county commissioner overseeing your clinic is an epidemiologist and wants to know how you plan to address the emergence of ciprofloxacin resistance in Neisseria gonorrhoeae Species of bacteria that is the causative agent of gonorrhea, a sexually transmitted infection

11. The Problem You run a public health clinic in Racine, Wisconsin A county commissioner overseeing your clinic is an epidemiologist and wants to know how you plan to address the emergence of ciprofloxacin resistance in Neisseria gonorrhoeae State budget cuts mean you cannot afford to give all of your patients more expensive antibiotics or do all of the lab tests that you would like

12. The Problem You run a public health clinic in Racine, Wisconsin A county commissioner overseeing your clinic is an epidemiologist and wants to know how you plan to address the emergence of ciprofloxacin resistance in Neisseria gonorrhoeae State budget cuts mean you cannot afford to give all of your patients more expensive antibiotics or do all of the lab tests that you would like  Develop a plan to address the medical, economic, and political questions your clinic will face in dealing with this public health issue

13. Sexually transmitted infection caused by the bacterium Neisseria gonorrhoeae Among the most common sexually transmitted infections (STI) in the world What is gonorrhea?

14. Gonorrhea is treated with antibiotics Antibiotic: chemical substance produced by or derived from a microorganism (molds or bacteria) that kills or inhibits the growth of bacteria Mold colony (Penicillium chrysogenum) Bacteria

15. Gonorrhea is treated with antibiotics Antibiotics help cure infections by decreasing the bacterial population to a level that the human immune system can handle Antibiotic treatment

16. Antibiotics disrupt essential cell processes

19. Outer cell membrane Cell wall Inner cell membrane + antibiotic (e.g. Penicillin) Cell wall construction

20. Antibiotics disrupt essential cell processes Cell wall construction

21. Antibiotics disrupt essential cell processes RNA DNA Protein Gene expression

22. Antibiotics disrupt essential cell processes RNA DNA Enzymes Cell structure Signaling Protein Gene expression

23. DNA replication occurs prior to cell division DNA replication Parent Cell Daughter Cells DNA

24. Antibiotics disrupt essential cell processes Replication DNA Gene expression

25. Antibiotics disrupt essential cell processes Transcription Replication DNA Gene expression RNA

26. Antibiotics disrupt essential cell processes Transcription Translation Replication DNA Gene expression RNA Protein

27. Antibiotics disrupt essential cell processes Transcription Translation Replication “Central Dogma of molecular biology” RNA Protein DNA Gene expression

28. Antibiotics disrupt essential cell processes Transcription Translation Replication + antibiotic RNA Protein DNA

29. Antibiotics disrupt essential cell processes Transcription Translation Replication + antibiotic RNA Protein DNA

30. Antibiotics disrupt essential cell processes Transcription Translation Replication + antibiotic RNA Protein DNA

31. Antibiotics disrupt essential cell processes Transcription Translation Replication + antibiotic Gene expression RNA Protein DNA

32. What is antibiotic resistance? The ability of bacteria to survive exposure to antibiotic drugs Antibiotic treatment

33. Resistance through alteration of drug target Enzyme active site antibiotic Wild type protein Mutant protein

34. Resistance through alteration of drug target antibiotic Wild type protein Mutant protein antibiotic Enzyme active site

35. DNA mutation can alter protein structure Wild typeMutant RNA Protein DNA

36. Genotype codes for the phenotype Every living organism is the outward physical manifestation of internally coded, inheritable, information

37. Genotype codes for the phenotype Every living organism is the outward physical manifestation of internally coded, inheritable information Genotype: The genetic constitution of an organism

38. Genotype codes for the phenotype Every living organism is the outward physical manifestation of internally coded, inheritable, information Genotype: The genetic constitution of an organism Phenotype: The physical features of an organism produced by the interaction of genotype and environment

39. Genotype codes for the phenotype RNA Protein DNA Genotype Phenotype Antibiotic Sensitivity antibiotic (Wild type)

40. Genotype codes for the phenotype RNA Protein DNA Genotype Phenotype Antibiotic Resistance antibiotic (Mutant)

41. The following statements describe how a change in genotype can perturb phenotype. Number them in the best sequential order (1=earliest event, 4=latest event): __ Mutant RNA is translated. __ Mutant DNA is transcribed. __ Mutant protein is altered, changing an organism’s response to its environment. __ An error occurs during DNA replication, resulting in a gene mutation. Genotype can perturb phenotype

42. The following statements describe how a change in genotype can perturb phenotype. Number them in the best sequential order (1=earliest event, 4=latest event): _3_ Mutant RNA is translated. _2_ Mutant DNA is transcribed. _4_ Mutant protein is altered, changing an organism’s response to its environment. _1_ An error occurs during DNA replication, resulting in a gene mutation. Genotype can perturb phenotype

43. Emergence of antibiotic resistance “It is not difficult to make microbes resistant to penicillin in the laboratory by exposing them to concentrations not sufficient to kill them, and the same thing has occasionally happened in the body.” - Alexander Fleming, 1945 Nobel Prize lecture

44. Gonococcal Isolate Surveillance Project (GISP) — Percent of Neisseria gonorrhoeae isolates with resistance or intermediate resistance to ciprofloxacin, 1990–2005 CDC STD Surveillance, 2005 Increasing resistance to antibiotics in US

45. Increasing resistance to antibiotics @ UW UHS Gonorrhea Isolation - 2006 –26 total cases Cervical/Vaginal (3) Urethral/Urine (14) Pharynx (4) Rectal (5) –9/9 non-genital isolates tested were ciprofloxacin resistant

46. Evolution of antibiotic resistance +++ Variation HeredityTime Selective Pressure

47. http://evolution.berkeley.edu Evolution of antibiotic resistance Variation

48. http://evolution.berkeley.edu Evolution of antibiotic resistance Variation Selective Pressure

49. http://evolution.berkeley.edu Evolution of antibiotic resistance Variation Heredity Selective Pressure

50. http://evolution.berkeley.edu Evolution of antibiotic resistance Variation HeredityTime Selective Pressure

51. Why aren’t antibiotics as effective as they used to be? What’s wrong with this statement?

52. 1. People have become immune to antibiotics –Human immune systems do not build up immunity to antibiotics. Rather, it is the bacteria within the human host that become resistant. What’s wrong with this statement?

53. 1. People have become immune to antibiotics –Human immune systems do not build up immunity to antibiotics. Rather, it is the bacteria within the human host that become resistant. 2. Antibiotics are changing and subsequently becoming weaker –Antibiotics are inert chemical substances. Bacteria, however, are living organisms that can change through evolution. What’s wrong with this statement?

54. 1. People have become immune to antibiotics –Human immune systems do not build up immunity to antibiotics. Rather, it is the bacteria within the human host that become resistant. 2. Antibiotics are changing and subsequently becoming weaker –Antibiotics are inert chemical substances. Bacteria, however, are living organisms that can change through evolution. 3. Bacteria have mutated in order to become resistant –Bacteria do not “choose” to mutate, adapt, or develop resistance. Natural selection selects among whatever variation exists in the population and the result is evolution. What’s wrong with this statement?

55. 1. People have become immune to antibiotics –Human immune systems do not build up immunity to antibiotics. Rather, it is the bacteria within the human host that become resistant. 2. Antibiotics are changing and subsequently becoming weaker –Antibiotics are inert chemical substances. Bacteria, however, are living organisms that can change through evolution. 3. Bacteria have mutated in order to become resistant –Bacteria do not “choose” to mutate, adapt, or develop resistance. Natural selection selects among whatever variation exists in the population and the result is evolution. 4. Antibiotics have mutated bacteria to become resistant –Antibiotics do not introduce mutations. Mutations occur randomly during DNA replication. What’s wrong with this statement?

56. Delaying the inevitable… 1.Don’t use antibiotics to treat viral infections. 2.Avoid mild doses of antibiotics over long time periods. 3.When treating a bacterial infection with antibiotics, take all of your pills. 4.Use a combination of drugs to treat a bacterial infection. 5.Reduce or eliminate the “preventative” use of antibiotics on livestock and crops. http://evolution.berkeley.edu