The document discusses soil sample collection by multiple students for the purpose of isolating bacteriophages. Gustavo collected soil samples from rural areas near bodies of water and vegetation at depths of 2-10 cm when temperatures were below 30 degrees Celsius. Juan collected samples from both urban and rural areas in Puerto Rico from parks, near trees and buildings at superficial depths when temperatures were below 30 degrees Celsius. Alejandra collected a dry sample near the coast in front of the Capitol in San Juan at 31 degrees Celsius. Angelin collected a damp sample from underneath grass at a university at 22 degrees Celsius. The goal is to find diverse mycobacteriophages near populated areas.

1. Juan E. Maldonado Weng
Alejandra González Torres
Gustavo A. Martínez Muñiz
Angelín M. Figueroa

2. Viruses that infect bacteria
Similar to fingerprints, no identical phages
have been identified
Great example of evolution and diversity
“Natural predators” of bacteria
So many, we want to analyze them all

3. Types of Reproduction
Lytic
Lysogenic
Lifestyles:
Lytic
Temperate

4. http://diverge.hunter.cuny.edu/~weigang/Images/13-12_lysogeniccycle_1.jpg

5. http://faculty.ccbcmd.edu/courses/bio141/lecguide/uni
t3/viruses/pen_lyt.html
http://faculty.ccbcmd.edu/courses/bio141/lecguide/un
it3/viruses/lysosum.html

6. Sterilize conical tube
Soil collection from different cities
Spoon to transfer the soil sample in the conical
tube
Fill the sterilized tube
Data Recorded
Date and time
Air temperature
Location
Depth
Moisture content
Features of the site

7. Enrichment
Filtration
Sterilize filter
Tuberculin syringes
Pipettes
Plaque screening
Mycobacterium smegmatis culture tubes
Positive control
Phage buffer
Top agar

8. Purification (looking for phages)
Three purification
Mycobacterium smegmatis culture
Agar plate
Plaque streak
Titer assay
To determine plaque-forming units concentration

10. 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

11. Gustavo Collection #1
Date: 02/18/2013
Time 17:44/5:44 P.M.
GPS Latitude: 18.21398
GPS Longitude: -65.995476
Air temp (approx.): 26.7 degrees Celsius
Depth: 2-2.5 cm
Moisture content (approx.): dry/moist
Features
Rural
Next to water tank cement box, and ficus roots
Not sloped.

12. Gustavo Collection #2
Date: 02/28/2013
Time: 16:13/4:13 P.M.
GPS Latitude: 18.221680
GPS Longitude: -65.996278
Air temperature (approx.): 30 degrees Celsius
Depth: 3 cm
Moisture content: saturated
Features
Rural
In a small field close to a river
Flat soil
No constructions nearby

13. Gustavo Collection #3
Date: 03/10/2013
Time: 18:25/6:25 P.M.
GPS Latitude: 18.215628
GPS Longitude: -65.995694
Air temperature (approx.):/ 22.2 degrees Celsius
Depth: 10 cm
Moisture content: damp and cool
Features
Rural area
In front of a house

14. Gustavo Collection #4
Date: 03/17/2013
Time: 19:16/7:16 P.M.
GPS Latitude: 18.218559
GPS Longitude: -65.998019
Air temperature (approx.): 26.1 degrees Celsius
Depth: 1 cm
Moisture content: moist and cool
Features
Rural area
In a canal
Next to a street

15. 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
5
03/17/201
3
1:59 P.M.
27 Superficial Moist Urban 18.221876 - 65.432118

16. Juan Collection #1
Date: 02/09/2013
Time 6:06 P.M.
GPS Latitude: 18.221986
GPS Longitude:-65. 432580
Air temp (approx.): 28 degrees Celsius
Depth: Superficial level
Moisture content (approx.): dry-moist
Features
A park in urban area
Close to several trees
Luquillo, Puerto Rico

17. Juan Collection #2
Date: 02/18/2013
Time 12:03 P.M.
GPS Latitude: 18.221952
GPS Longitude:- 65.432384
Air temp (approx.): 27 degrees Celsius
Depth: Superficial level
Moisture content (approx.): dry-moist
Features
A park in urban area
Close to several trees
Luquillo, Puerto Rico

18. Juan Collection #3
Date: 03/03/2013
Time 3:29 P.M.
GPS Latitude: 18.221934
GPS Longitude: -65.43166
Air temp (approx.): 29 degrees Celsius
Depth: Superficial level
Moisture content (approx.): dry
Features
Urban area near a cable company
Under a tree
Luquillo, Puerto Rico

19. Juan Collection #4
Date: 03/11/2013
Time 6:36 P.M.
GPS Latitude: 18.7575
GPS Longitude: -66.84186
Air temp (approx.): 27 degrees Celsius
Depth: Superficial level
Moisture content (approx.): dry
Features
Country side
Elevated area
Cayey, Puerto Rico

20. Juan Collection #5
Date: 03/17/2013
Time 1:59 P.M.
GPS Latitude: 18.221876
GPS Longitude: - 65.432118
Air temp (approx.): 27 degrees Celsius
Depth: Superficial level
Moisture content (approx.): moist
Features
Urban Area
Backyard
Luquillo, Puerto Rico

21. Trial Name: Mirificus tempus
Collected at San Juan, Puerto Rico
Time 12:45 P.M.
GPS Latitude: 18 28'2.96” N
GPS Longitude: 66 6'23.92” W
Air temp (approx.): 31.1 degrees Celsius
Depth: Superficial level
Moisture content (approx.): dry

22. Enrichment:
Stimulates the growth of phages with bacteria
Plaque Purification:
We need our phages as clean as possible to be able
to study them.

28. Gustavo - 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.

29. 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.

30. 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 coast
At an approximate temperature of 31

31. Angelin - Samples were gathered:
At the university’s athlete housing at Cayey
Late at night
Damp moisture content; 2 cm underneath the grass
At temperature approximately 22 degrees Celsius.

32. Mycobacteriophages
To best find one:
Near the coast
Near a busy city
Many more people
Finding more phages will lead to more concrete
answers

33. 42
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