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Phage presentation- The Isolation and Characterization of Mycobacteriophage Musamodel from Tropical Soils of Puerto Rico- spring 2014
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Phage presentation- The Isolation and Characterization of Mycobacteriophage Musamodel from Tropical Soils of Puerto Rico- spring 2014

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  • 1. ISOLATION OF A NOVEL MYCOBACTERIOPHAGE Lizbeth Pérez Castro Javier Zavala Ayala Mónica Del Moral Félix Vallés
  • 2. WHAT IS A BACTERIOPHAGE? • A bacteriophage is any virus that infects any bacterium. • Mycobacteriophages are viruses that infect mycobacteria. • Found in different bacterial colonies. • Mostly found in the soil and sea water. • Resistant to antibiotics. http://news.bbcimg.co.uk/media/images/66413000/ jpg/_66413229_m0900092-bacteriophage_virus-
  • 3. BACTERIAS FOR HOSTING 1. M. smegmatis (Smeg) • fast growing rate and non-pathogenic behaviour • Aerobic organisms • Found in water, soil and plants 2. Bacillus • Obligate aerobes or facultative anaerobes • Well understood- used as a model for differentiation, gene regulation, and cell cycle events in bacteria. • Common in soil  Both are:  Gram positive  Acid fast bacteria  Morfology- rodshape
  • 4. BACTERIOPHAGE STRUCTURE • Phages consist of 3 basic components: • Capsid-contains the genetic material • Genetic material- DNA • Tail-serves to attach to bacteria and DNA passes through tail and into bacterium http://en.wikipedia.org/wiki/Bacteriophage
  • 5. PHAGE LIFE CYCLES • Temperate-Lysogenic: • Replicate and lyse or dormant state- incorporate its genetic material into DNA of host. • Virulent-Lytic: • Straightforward cycle by lysing all bacteria they infect.
  • 6. http://biomedfrontiers.org/infection-2014-3-28/
  • 7. INTRODUCTION • Problem: Can novel bacteriophages be isolated from tropical soil of Puerto Rico? • Hypothesis: Unique bacteriophages will be isolated and characterized from a collected soil sample of Puerto Rico. http://news.bbcimg.co.uk/media/images/66413000/jpg/_664 13229_m0900092-bacteriophage_virus-spl.jpg
  • 8. PROCEDURE httpwww.teachthechil drenwell.comscience.h tml http://www.exportersin dia.com/swastikaelectr ic/products.htm http://quizlet.com/6760545/lab-3-streak-plate- technique-flash-cards/ Soil Enrichment Filtration Purification HTPLTen Plate Preparation
  • 9. MY BACTERIOPHAGE • Name: Nitidusvenutus • Located in: Carretera 943 Sector Los Chinos, Gurabo • Coordinates: 18º 16’ 7’ N, 65º 58’ 32’ W • Collected: February 24, 2014 • It was found in a rural area next to cow feces on a template day.
  • 10. LOCATION
  • 11. MY BACTERIOPHAGE
  • 12. RESULTS http://faculty.plattsburgh.edu/don ald.slish/lambdatitre.html
  • 13. MY BACTERIOPHAGE: MUSAMODEL • Name- Musamodel • Located in: 1-9 calle Barcelona, Urb.Terralinda, Caguas P.R • Coordinates: 88˚14’6”N , 66˚2’39”W • Collected: March 16, 2014 • Time: 7:51pm • Temperature: 79˚ • About 3 inches deep and 1 foot close to concrete • Found next to a plantain tree • Soil was moist and loose
  • 14. LOCATION
  • 15. SAMPLES COLLECTED 1st 2nd 3rd 4th 5th 6th 7th Date 2/6/14 2/18/14 2/24/14 3/3/14 3/10/14 3/16/14 3/16/14 Time 7:53 am 7:06 am 9:55 pm 9:38 am 6:26 am 7:45 pm 7:51 pm Coordinates 18˚14’6”N, 66˚2’40”W 18˚14’13”N, 66˚2’35”W 18˚14’7”N, 66˚2’39”W 18˚14’18”N, 66˚2’42”W 18˚14’7”N, 66˚2’40”W 18˚14’6”N, 66˚2’39”W 18˚14’17”N, 66˚2’42”W Temperature 75˚ 75˚ 79˚ 81˚ 73˚ 79˚ 79˚ Soil description Moist, 3 inches deep Moist, 1 inch deep Moist, 1 inch deep Moist, with moss, 1 inch deep Moist, 1 inch Moist, loose, 5 inches deep Moist, loose, 3 inches deep Site description Urban, 5 feet close to concrete Urban, 1 foot from concrete Urban, 2 feet from concrete Urban, ½ foot from concrete Urban, 3 feet away from concrete Urban, 1 foot close to concrete Next to a plantain plant, 1 foot close to concrete
  • 16. RESULTS
  • 17. CONCLUSION • Two mycobacteriophages were found: 1. Nitidusvenutus a. Located in Carretera 943 Sector Los Chinos, Gurabo. b. Based on the clear plaques, we can assume it has a lytic life cycle. 2. Musamodel a. Located in 1-9 calle Barcelona, Urb. Terralinda, Caguas P.R. b. Up to now, we believe it has a lytic life cycle because of clear plaques. c. Three purifications were completed.
  • 18. FUTURE PLANS • Nitidusvenutus; • Isolation of the bacteriophage’s DNA • Bioinformatics analyses and sequencing. • Musamodel; • 3rd purification- to have more isolated plaques and measure diameter • Spot test • Make phage stocks- • Empirical test • 10 plate preparation • Analyze • Bioinformatics and sequencing
  • 19. ACKNOWLEDGEMENT • Dr. Michael Rubin • Juan Apiz • Eduardo Correa • Gustavo Martínez • Giovanni Cruz • Edwin Alvarado