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Mycobacteriophages:The Journey
Juan E. Maldonado WengAlejandra González TorresGustavo A. Martínez MuñizAngelín M. Figueroa
The Beginning:Mycobacteriaphages
Viruses that infect bacteriaSimilar to fingerprints, no identical phages have been identifiedGreat example of evolution and diversity“Natural predators” of bacteriaSo many, we want to analyze them all
The Beginning:The Reproduction
Types of ReproductionLyticLysogenic
Lifestyles:LyticTemperate
The Beginning:The Diagrams
http://diverge.hunter.cuny.edu/~weigang/Images/13-12_lysogeniccycle_1.jpg
The Beginning:The Diagrams
http://faculty.ccbcmd.edu/courses/bio141/lecguide/unit3/viruses/pen_lyt.html
http://faculty.ccbcmd.edu/courses/bio141/lecguide/unit3/viruses/lysosum.html
The Plan:The Collection
Sterilize conical tubeSoil collection from different citiesSpoon to transfer the soil sample in the conical tubeFill the sterilized tube Data Recorded
Date and time Air temperatureLocationDepthMoisture contentFeatures of the site
The Plan:Materials and Methods
EnrichmentFiltration
Sterilize filterTuberculin syringesPipettes
Plaque screeningMycobacterium smegmatis culture tubesPositive controlPhage bufferTop agar
The Plan:Materials and Methods
Purification (looking for phages)Three purificationMycobacterium smegmatis cultureAgar plate
Plaque streakTiter assay
To determine plaque-forming units concentration
The Plan:The Strategy
The Data
Gustavo's Collections
Collection # Date Time Air Temp. (Celsius) Depth Moisture Features GPS Latitude GPS Longitude
1 2/18/2013 5:44 P.M. 26.7 2-2.5 cm Dry/Moist Rural 18.21398 -65.995476
2 2/28/2013 4:13P.M. 30 3cm Saturated Rural 18.22168 -65.996278
3 3/10/2013 6:25 P.M. 22.2 10 cm Damp Rural 18.215628 -65.995694
4 3/17/2013 7:16 P.M. 26.1 1 cm Moist Rural 18.218559 -65.998019
The Data
Gustavo Collection #1Date: 02/18/2013Time 17:44/5:44 P.M.GPS Latitude: 18.21398GPS Longitude: -65.995476Air temp (approx.): 26.7 degrees CelsiusDepth: 2-2.5 cmMoisture content (approx.): dry/moistFeatures
RuralNext to water tank cement box, and ficus rootsNot sloped.
The Data
Gustavo Collection #2Date: 02/28/2013Time: 16:13/4:13 P.M.GPS Latitude: 18.221680 GPS Longitude: -65.996278Air temperature (approx.): 30 degrees CelsiusDepth: 3 cmMoisture content: saturatedFeatures
RuralIn a small field close to a riverFlat soil No constructions nearby
The Data
Gustavo Collection #3Date: 03/10/2013Time: 18:25/6:25 P.M.GPS Latitude: 18.215628GPS Longitude: -65.995694Air temperature (approx.):/ 22.2 degrees CelsiusDepth: 10 cmMoisture content: damp and coolFeatures
Rural areaIn front of a house
The Data
Gustavo Collection #4Date: 03/17/2013Time: 19:16/7:16 P.M.GPS Latitude: 18.218559GPS Longitude: -65.998019Air temperature (approx.): 26.1 degrees CelsiusDepth: 1 cmMoisture content: moist and coolFeatures
Rural areaIn a canalNext to a street
Juan's Collections
Collection # Date Time Air Temp.
(Celsius) Depth Moisture Features GPS Latitude GPS Longitude
1 2/9/2013 6:06 P.M. 28 Superficial Dry/Moist Urban 18.221986 -65. 432580
2 2/18/2013 12:03 P.M. 27 Superficial Dry-moist Urban 18.221952 - 65.432384
3 3/3/2013 3:29 P.M. 29 Superficial Dry Urban 18.221934 - 65.43166
4 3/11/2013 6:36 P.M. 27 Superficial Dry Urban 18.7575 - 66.84186
503/17/201
3 1:59 P.M. 27 Superficial Moist Urban 18.221876 - 65.432118
The Data
The Data
Juan Collection #1Date: 02/09/2013Time 6:06 P.M.GPS Latitude: 18.221986GPS Longitude:-65. 432580Air temp (approx.): 28 degrees CelsiusDepth: Superficial levelMoisture content (approx.): dry-moistFeatures
A park in urban areaClose to several treesLuquillo, Puerto Rico
The Data
Juan Collection #2Date: 02/18/2013Time 12:03 P.M.GPS Latitude: 18.221952GPS Longitude:- 65.432384Air temp (approx.): 27 degrees CelsiusDepth: Superficial levelMoisture content (approx.): dry-moistFeatures
A park in urban areaClose to several treesLuquillo, Puerto Rico
The Data
Juan Collection #3Date: 03/03/2013Time 3:29 P.M.GPS Latitude: 18.221934GPS Longitude: -65.43166Air temp (approx.): 29 degrees CelsiusDepth: Superficial levelMoisture content (approx.): dryFeatures
Urban area near a cable companyUnder a treeLuquillo, Puerto Rico
The Data
Juan Collection #4Date: 03/11/2013Time 6:36 P.M.GPS Latitude: 18.7575GPS Longitude: -66.84186Air temp (approx.): 27 degrees CelsiusDepth: Superficial levelMoisture content (approx.): dryFeatures
Country sideElevated areaCayey, Puerto Rico
The Data
Juan Collection #5Date: 03/17/2013Time 1:59 P.M.GPS Latitude: 18.221876GPS Longitude: - 65.432118Air temp (approx.): 27 degrees CelsiusDepth: Superficial levelMoisture content (approx.): moistFeatures
Urban AreaBackyardLuquillo, Puerto Rico
Mycobacteriaphage:The Find
Trial Name: Mirificus tempusCollected at San Juan, Puerto Rico Time 12:45 P.M.GPS Latitude: 18 28'2.96” NGPS Longitude: 66 6'23.92” WAir temp (approx.): 31.1 degrees CelsiusDepth: Superficial levelMoisture content (approx.): dry
The Second part of the Plan:Methodology
Enrichment:Stimulates the growth of phages with bacteria
Plaque Purification: We need our phages as clean as possible to be able to study them.
The ResultsGustavo - Samples were gathered:
Dark brown or near black soil; damp to moist; around or under vegetation and near bodies of water. In a mountainous, rural area. From a closed-access neighborhood which had been constructed 10-15 years earlier.Near houses or man-made constructions.At temperatures equal to or below 30 degrees Celsius.
Juan - Samples were gathered:Dry to moist soil; near or under trees; one sample was from rocky soil. In an urban area, then rural area. In parks, near houses or cable company.At temperatures equal to or below 30 degrees Celsius.
The Results
Alejandra - Samples were gathered:Loose, dry soil; in grass vegetation; In an urban area, in front of the Capitol of P.R. Close to the coastAt an approximate temperature of 31
The Results
Angelin - Samples were gathered:At the university’s athlete housing at CayeyLate at nightDamp moisture content; 2 cm underneath the grassAt temperature approximately 22 degrees Celsius.
The Results
The End of the Journey:The conclusions
MycobacteriophagesTo best find one:
Near the coastNear a busy cityMany more people
Finding more phages will lead to more concrete answers
The End
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