ISOLATION OF A NOVEL MYCOBACTERIOPHAGE

Lizbeth Pérez Castro

Javier Zavala Ayala

Mónica Del Moral

Félix Valles

RISE Program UPR Cayey

WHAT IS A BACTERIOPHAGE?• A bacteriophage is any virus that infects any bacterium.

• Mycobacteriophages are viruses that infect mycobacteria.

• All mycobacteriophages are bacteriophages, but not all bacteriophages are mycobacteriophages.

• Found in different bacterial colonies.

• Mostly found in the soil and sea water.

• Resistant to antibiotics.

BACTERIAS FOR HOSTING1. 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

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

PHAGE LIFE CYCLES

• Temperate-Lisogenic:

• Replicate and lyse or dormant state- incorporate its genetic material into DNA of host.

• Virulent-Lytic:

• Straightforward cycle by lysing all bacteria they infect.

http://biomedfrontiers.org/infection-2014-3-28/

PROCEDURE

httpwww.teachthechildrenwell.comscience.

html

http://www.exportersindia.com/swastikaelectric/products.htm

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

Soil

EnrichmentFiltration

Purification

HTLTen Plate Preparation

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.

LOCATION

MY BACTERIOPHAGE

RESULTS

http://faculty.plattsburgh.edu/donald.slish/lambdatitre.html

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

LOCATION

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

RESULTS

CONCLUSION

• Two mycobacteriophages were found:

1. Nitidusvenutusa. located in Carretera 943 Sector Los Chinos, Gurabob. ……. Were completed

2. Musamodela. located in 1-9 calle Barcelona, Urb. Terralinda, Caguas P.Rb. Up to now, we believe it has a lysogenic life cycle because of turbidnessc. three purifications were completed

FUTURE PLANS

• Nitidusvenutus

• 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

ACKNOWLEDGEMENT

• Dr. Michael Rubin

• Juan Apiz

• Eduardo Correa

• Gustavo Martínez

• Giovanni Cruz