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
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http://quizlet.com/6760545/lab-3-streak-plate-
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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. 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.

14. FUTURE PLANS
• Nitidusvenutus;
• Isolation of the bacteriophage’s DNA
• Bioinformatics analyses and sequencing.

15. ACKNOWLEDGEMENT
• Dr. Michael Rubin
• Juan Apiz
• Eduardo Correa
• Gustavo Martínez
• Giovanni Cruz
• Edwin Alvarado