presentation bacteriophages
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Isolation, Purification and Characterization of Gêmeos and
Isinaayusferti, bacteriophages Isolated from Tropical Soils
Amanda M. Icazatti Burtell, Fernando Pachecho Ocasio, John Rivera Meléndez & Leyda Villagrasa Méndez
Dr. Michael RubinRISE Program
University of Puerto Rico at Cayey
Introduction
• What is a bacteriophage?• What host bacteria are being used?– Mycobacterium smegmatis– Bacillus cereus
• Where can bacteriophages be found?• What do bacteriophages need for living?• Types of life cycles: Lytic vs. Lysogenic cycle
• Why are bacteriophages important?Phages can be used in various fields of science such
as: - Genetics - Cloning and mutation of DNA and
studies in genome- Therapeutic uses for the creation of antibiotics resistant to bacteria
• What is the purpose of this experiment?
Procedure
Collection of SoilEnrichment Centrifuge
FiltrationStreakin Process: Purification X3
DilutionsSpot test
Empirical test
Web pattern
Preliminary Results
Sample Coordinates Details Depth Temperature Date Place
Sample #1AMIB
Lat: 18.21561L:on: -66.044790
Humid and Dry soil
3’’ 25˚C 02/02/14 Caguas, PR
Sample # 2AMIB
Lat: 18.234378Lon:-66.061491
Dry soil 2’’ 25˚C 02/17/14 Caguas, PR
Sample #3AMIB
Lat: 18.234604Lon:-66.06138
Humid and Dry soil
1’’ 27˚C 02/24/14 Caguas, PR
Sample #4AMIB
Lat: 18.213103Lat: -66.041709
Humid and Damp soil ’
3’’ 26˚C 03/04/14 Caguas, PR
Sample #5AMIB
Lat: 18.215715Lon: -66.044879
Dry soil 1’’ 25˚C 03/11/14 Caguas, PR
Sample #1FPO
CoordinatesLat: 18.11’15”NL:on: 65.57’54”
DetailsDry soil
Depth3”
Temperature24c
Date04/02/14
Place San Lorenzo, Pr
Sample #2FPO
Lat: 18.11’15”NLon: 65.57’53”W
Very dry soil.
4” 26˚C 18/02/14 San Lorenzo, Pr
Sample #3FPO
Lat: 18.7’6”NLon: 66.9’42”W
Humind Soil
3” 28˚C 18/02/14 Cayey, Pr
Sample #4FPO
Lat: 18.11’15”NLon: 65.57’53”W
Humid Soil
4” 22˚C 26/02/14 San Lorenzo, Pr
Sample #5FPO
Lat: 18.2’26”NLon: 66.35’35”
Humid Soil
6” 30˚C 02/03/14 Ponce, Pr
Gêmeos (bacilophage)• Coordenates: Lat: 23' 36" 23 S Long: 46' 41" 51 W• Temperature: 19˚C• Details: Dark and damp soil • Found in a populated area near a river• Sao Paolo. Brazil
Gêmeos: Preliminary Results
Sample Coordenates Details Depth Temperature Date Place
Sample #1LVM
Lat: 18.2333419Long:-66.0478104
Humid and plain soil
2” 26˚C Feb/3/14 Caguas, PR
Sample # 2LVM
LAT: 18.1192448LONG: -66.1667764
Dry and lumpy soil
1” 25˚C Feb/16/14
Cayey, PR
Sample #3LVM
Lat: 18.2333419Long:-66.0478104
Humid and plain soil
3” 27˚C Feb/23/14
Caguas, PR
Sample Coordenates Details Depth Temperature Date Place
Sample #1JLR
Lat: 18ᵒN Long: 66ᵒW
Humid and saturated soil
½ ‘’ 31˚C Feb/3/14 Cidra, PR
Sample # 2JLR
Lat: 18ᵒNLong: 66ᵒW
Dry soil under trash disposal
Surface sample
32˚C Feb/5/14 Cidra, PR
Isinaayusferti (mycobacteriophage)Coordenates: Lat: 18˚ 14’ .0312 " N Long: 66˚ 2' 52.116" WTemperature: 27 ˚ C Place: Caguas, Puerto RicoDetails: Humid and plain soil. Found near the garbage in front of a house.
Isinaayusferti: Preliminary Results
Conclusion
• The soil samples were collected and isolated using the enrichment method.
• A bacilophage was found in Sao Paolo, Brazil and was named Gêmeos.
• Third Purification Stock Filtrate (Gêmeos)• A mycobacteriophage found in Caguas, Puerto
Rico and was named Isinaayusferti.• Concentrated Stock Phage (Isinaayusferti)
Acknoledgements
• RISE Program• Dr. Michael Rubin• Dr. Eneida Díaz• Gustavo Martínez• Giovanni Cruz• Christopher Quintanal
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