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Mycobacteriophages presentation
 

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    Mycobacteriophages presentation Mycobacteriophages presentation Presentation Transcript

    • MycobacteriophagesBy:Anaeli ShockeyNicolle A. RosaDelaine M. Zayas-BazanZuleika Velázquez OrtizBIOL 4997Rise Program
    • Introduction• Mycrobateriophages are viruses that infectbacteria belonging to the Mycibacterium genus.• They are relatively easy to isolate andcharacterize.• They are great for studying evolution.• Bacteriophages have two cycles: lytic cycle(reproduction) and lysogenic cycle (integration)
    • • In our experiment we looked formycobacteriophages and completed the requiredsteps to isolate and characterize it.• This had some implications such as: Learning the Aseptic Technique Applying other laboratory skills: plating,centrifugation and mixing solutions among others. Understanding the importance of finding andcharacterizing a mycobacteriophage
    • Materials Sample Collection Lab Notebook Sterile 15mL conical tube with cap Sterile Spoon GPS or Google Earth
    • Materials Enrichment Culture Clean Spatula Approximately 0.5g of the soil sample Sterile 50mL conical tube with cap Weighing pan ----muestra
    • Materials Harvesting and Plaque Screening 5mL sterile disposable pipettes Agar Plates Bacteria Centrifuge Enriched Sample Sterile 15mL conical tube with cap Sterile Filter Sterile Syringe Tips Top Agar
    • Methods Sample Collection Identify the place for soil sample collection Recollect the soil sample with a sterile spoonand put it in the sterile 15mL conical tube withcap Close quickly Take two picture (place and you) and theenvironmental data.
    • Methods Enrichment Culture Obtain ~0.5g of the soil sample Added it to the … solution placed in the sterile50mL conical tube with cap. Close it slightly Incubate at 37° C for 24 hours
    • Methods Harvesting and Preparing theEnriched Sample Centrifuge the enriched sample at 3000 RPM for 10minutes Prepare phage filtrate Obtain the supernatant of the sample Filter the supernatant into a sterile 15mL conical tubewith cap and close quickly
    • Method Plaque Screening Label the Agar Plate with the necessary information andname the zone of the Agar Plate with number (1, 2 and 3) In the extracted supernatant wet the wood side of a tip. Make the first striate with the wet tip in the zone 1 Use another tip to disperse the striate of the zone 1 to thezone 2 and the same procedure with the zone 2 to thezone 3.
    •  Add 5.0mL of Top Agar with the bacteria to theAgar Plate from the zone with the less amount ofsample to the most amount of sample Let the Top Agar solidifies, invert the plate quickly. Incubate the inverted plate at 37°CObservePhages or Not Phages?
    • Data
    • Date Time Weather Location OtherConditions GPS InformationTemperature: 28°C Site: Urban Depth: .5cmPrecipitation: 10% Proximity to: Trees This soil sample wasFebruary 9th 2:45 PM Humidity: 65% and buildings not utilized due to theWind: 13km/h 18°0844.56"N 65°4917.55"W absence of bacteriaSunny/Clear Humacao, Puerto Rico in the lab.Temperature: 27.2°C Site: Urban Depth: .5cmPrecipitation: 40% Proximity to: Trees This soil sample wasFebruary 18th 1:39 PM Humidity: 69% and buildings utilized, but it did notWind: 20km/h 18°0844.56"N 65°4917.55"W contain phages.Cloudy Humacao, Puerto RicoTemperature: 29°C Site: Urban Depth: .3cmProximity to: Beach This soil sample wasMarch 3rd 1:00 PM Humidity: 67% and to the road utilized, but it did notWind: 24km/h 18°109.38"N 65°4428.82"W contain phages.Sunny/Clear Humacao, Puerto RicoTemperature: 31°C Site: Rural Depth: .5cmProximity to: Trees This soil sample wasMarch 3rd 4:36 PM Humidity: 49% and buildings utilized, but it did not18°175.66"N 66° 041.57"W contain phages.Sunny/Clear Gurabo, Puerto RicoTemperature: 27°C Site: Urban Depth: .5cmProximity to: Very close to This soil sample wasDelaine Soil Samples
    • Date Time Weather Location OtherConditions GPS InformationTemperature: 27°C Site: Urban Depth: .5cmProximity to: Very close to This soil sample wasMarch 11th 6:36 PM buildings utilized, but it did not18°75"N 66°843"W contain phages.Clear/ Dark Cayey, Puerto RicoTemperature: 29°C Site: Rural Depth: .2cmProximity to: Trees This soil sample wasMarch 17th 3:27 PM Humidity: 69% and buildings utilized, but it did not18°175.55"N 66° 041.29"W contain phages.Sunny/Clear Gurabo, Puerto RicoTemperature: 29°C Site: Rural Depth: .6cmProximity to: Trees This soil sample wasMarch 17th 3:30 PM Humidity: 69% and buildings utilized, but it did not18°1710.43"N 66° 039.28"W contain phages.Sunny/Clear Gurabo, Puerto RicoTemperature: 29°C Site: Rural Depth: .4cmProximity to: Trees This soil sample wasMarch 17th 3:32 PM Humidity: 69% and buildings utilized, but it did not18°175.79"N 66° 042.14"W contain phages.Sunny/Clear Gurabo, Puerto Rico
    • Zuleika Soil SampleDate Time Weather Condition LocationGPSOtherInformationFebruary 18 4:14 PMTemperature: 68°Cloudy/ ClearSite: RuralProximity to: 5 minutes tothe main road, trees and myhouse18° 1006.46"N66° 1130.52" WCidra, Puerto RicoDepth: 1cmThis soil sample wasutilized, but it did notcontain phages.March 3 1:19 PMTemperature: 76°Sunny/ClearSite: RuralProximity to: 5 minutes tothe main road, trees and myhouseCidra, Puerto RicoDepth: 0.8cmThis soil sample wasutilized, but it did notcontain phages.March 12 9:36 AMTemperature: 79°Sunny/ClearSite: RuralProximity to: fruits treesand my houseCidra, Puerto RicoDepth: 1.2cmThis soil sample wasutilized, but it did notcontain phages.March 12 9:42 AMTemperature: 79°Sunny/ClearSite: RuralProximity to: fruits treesand my houseCidra, Puerto RicoDepth: 0.6 cmThis soil sample wasutilized, but it did notcontain phages.April 8 8:18 AMTemperature: 82°SunnySite: RuralProximity to: 5 minutes tothe main road, trees and myhouseCidra, Puerto RicoDepth: 0.7cmThis soil sample wasutilized, but a problem withthe bacteria was presented.It did not appear phages.
    • Anaeli and Nicolle Soil SampleDate Time Weather Location OtherConditions GPS InformationTemperature: 25.6°C Site: Urban Depth: 5.74 cmProximity to: Trees This soil sample was3:32 PMand compostutilized and resulted in morethan one phage.18°1448.53"N 66° 06.55"WSunny/Clear Gurabo, Puerto RicoDate Time Weather Condition LocationGPSOtherInformationFebruary 18 7:03 PMTemperature: 80°RainySite: UrbanProximity to:Street Dr. DiegoÁlvarez Chanca,LevittownToa Baja, Puerto RicoDepth: 3.4cmThe soil sample wasused, but the result wasnegative.
    • Comparison All of the samples were diverse Hours, locations, and temperatures among other determining factors Factors The municipalities They have varying geographic and geomorphic The blue represents the karst region The yellow represents the coastal region The green represents the inner mountain region
    •  Type of soil◦ Unfortunately this information was not adequatelyrecorded, because the only description mentioned was dry,moist or normal.◦ There are three types of soil: sand, clay and silt, and each ofthose contain certain characteristics that difference them.◦ Vegetation surrounding the location The vegetation is diverse from municipality tomunicipality.
    • Conclusion Although the experiments and procedures wereperformed accurately, no bacteriophages were found. This might be for any of the reasons discussed in thecomparison or because the laboratory technicalities,such as the availability of bacteria. The main objective was not accomplished, but otherobjectives like practicing important laboratory skillswere completed.