Antibiotic Resistant Bacteria at the Beach. Megan May Waquoit Bay Research Reserve April 30, 2015

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1 Antibiotic Resistant Bacteria at the Beach Megan May Waquoit Bay Research Reserve April 30, 2015

2 Outline My research background Background Microbes Antibiotics Antibiotic Resistance My research project Larger significance

3 Applied Environmental Microbiology Microbiology Public Health Aquatic Environments

4

5 Research Experiences

6 Graduate School MIT-WHOI Joint Program in Oceanography Dr. Rebecca Gast s Lab at WHOI

7 Background Information

8 What are microbes? Organisms that can be seen with a microscope Actinetobacter (Bacteria) Candida (Fungus, Eucarya) Sulfolobus (Archaea) Microbiology Online, CDC AR Report

9 Life is divided into 3 groups. Pace 2009

10 Life is Mainly Microbes! Pace 2009

11 Why do we care about microbes? Nutrient cycling, degradation Fermentation Bioremediation Bioprospecting

12 Why do we care about microbes? Hydrothermal vents Ocean sediments Yellowstone National Park Ice

13 Microbes are all around us Environment Amount Ocean Water 500,000 Cells/cm 3 Ocean sediments (top 10 cm) 220,000,000 cells/cm 3 Rivers 1,000,000 Cells/cm 3 Forest sediments 40,000,000 cells/gram Whitman et al. 1998

14 Microbes are all over us

15 Bacteria and archaea outnumber us by a lot! 4,000,000,000,000,000,000,000,000,000,000 7,310,000,000 Whitman et al. 1998

16 Disease-causing (pathogenic) microbes are not the majority Tuberculosis Blood poisoning Pneumonia Dysentery Pneumonia

17 Antibiotics

18 Discovery of antibiotics Alexander Fleming

19 Fleming s Work

20 Antibiotics compound produced by a microorganism that inhibits the growth of another microorganism natural process can be chemically synthesized Kummer 2009

21 Recent discovery by humans Wright 2007

22 Humans use antibiotics often. Animal Agriculture Aquaculture Human health Agriculture

23 How do antibiotics work? Targeting the bacterial cells to stop them from growing or to kill them Can occur by attacking the outside of the cell or by affecting the processes inside the cell

24 What is antibiotic resistance? Resistance is when a bacteria is not affected by a particular antibiotic Natural occurring process Found in ancient sediments Isolated caves Isolated tribe of humans

25 How can bacteria resist antibiotics? Change the target Pump out antibiotics Immunity Inactivate the activity Modified from: Wright 2010

26 How can resistance be spread? Selections by exposure to antibiotics Bacteria sharing resistance CDC Threat Report, 2013

27 Marine Environmental Resistance Resistance naturally occurs in the environment. The ocean acts as a reservoir for resistance. Human caused antibiotic inputs enter enviroment. Allows for resistance to be shared in the ocean environment.

28 AR transfer from the ocean Recreation Ocean Humans Consumption

29 Environment is important to understanding our health.

30 Antibiotic resistance has large impacts. CDC Threat Report, 2013

31 What are the levels of antibiotic resistance at beaches around Cape Cod?

32 Why the beach? Land-sea interface Humans interact Consumption Recreation

33 Why would we expect resistance in the environment? Natural process Likely enhanced by humans Resistance can enter through wastes Actual antibiotic residues can enter

34 OSB WB and NRB LI Buzzards Bay SCB BDN Nantucket Sound

35 Methods Flow Chart Sampling Culture Bacteria Resistance Testing

36 Field Sampling

37 Ice doesn t stop our sampling!

38 Methods Flow Chart Sampling Culture Bacteria Resistance Testing

39 Culturing Enterococcus Vibrio General Marine Bacteria Fecal indicator used By the EPA -Can be pathogenic Widespread in the Marine environment -Can be pathogenic Indicates main environmental reservoir

40 Picking isolates Enterococcus Vibrio General Marine Bacteria 500 isolates 500 isolates 1500 isolates

41 Methods Flow Chart Sampling Culture Bacteria Resistance Testing

42 Resistance Testing Ciprofloxacin Antibiotic Discs Trimethoprim Clindamycin Doxycycline Amoxicillin Inhibition Zone Oxytetracycline

43 Resistance Testing

44 Why these antibiotics? Antibiotic Selected Uses Use began Amoxicillin Ulcers, Lyme disease, bronchitis, ear infections 1972 Ciprofloxacin Clindamycin Doxycycline Oxytetracycline Trimethoprim Diarrhea, pneumonia, urinary tract infections Blood poisoning, pneumonia, malaria, acne Cholera, Gonorrhea, acne, diarrhea, Lyme disease Syphilis, Pneumonia, Bronchitis, plague, throat irritation Urinary tract infections, pneumonia s 1982

45 Resistance in cultured beach bacteria Enterococcus Vibrio General Marine Bacteria In process 94.3 % (316 isolates) ~50 more to be collected In process

46 What have we learned so far?

47 Number of Bacterial Isolates Resistance is prevalent within tested Vibrio Resistant to this Number of Antibiotics

48 Percent Resistance Preliminary seasonal pattern June July Aug Sep Oct Dec

49 Percent Resistance Site differences are not large LI OSB SCB WB NRB BDN Buzzards Bay Waquoit Bay

50 Percent Resistant (%) Antibiotic results vary s

51 Percent Resistance Sample type differences are small wet dry water

52 More resistance compared to other studies Other studies done in the developing world Tend to have smaller numbers Test different antibiotics

53 Future work Working on resistance testing Enterococcus General marine bacteria Identify bacteria by genetic analysis Quantification of human impacts

54 Where does this research fit into the larger picture?

55 Marine Resistance Study Goals Better understanding of the marine environment Understand if there are links to human impacts Determine health risks Determine if necessary to make changes in use

56 AR is a global and national problem.

57 What can you do?

58 Preserve the integrity of antibiotics Levy 1998

59 If able to, buy antibiotic free food

60 Enjoy the beach!

61 Acknowledgments Gast lab Rebecca Gast Roxanne Beinart Waquoit Bay National Estuarine Research Reserve Joan Mueller Chris Weidman Funding sources MIT WHOI Ocean Ventures Fund WHOI Academic Programs Office WHOI Coastal Ocean Institute National Science Foundational Graduate Research Fellowship Field sampling associates Rebecca Gast Gabriella Farfan Nat Wilson Jenny Wehof Net Charoenpong Alec Bogdanoff Hanny Rivera Erin Black Lauren Kipp MIT-WHOI Joint Program