PHAGE PHOLKS

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PHAGE PHOLKS

• VIRUSES

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GENETIC ELEMENTS

• VIRUSES ARE GENETIC ELEMENTS ENCLOSED IN PROTEIN
  AND ARE NOT CONSIDERED TO BE ORGANISMS, BECAUSE
  THEY CANNOT REPRODUCE INDEPENDENTLY.

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BACTERIOPHAGES

• BACTERIOPHAGES ARE VIRUSES THAT INFECT BACTERIA
• MANY BACTERIOPHAGES, CALLED PHAGES FOR SHORT, ARE
  LARGE AND COMPLEX WITH LARGE AMOUNTS OF DNA AND
  PROTEINS.

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ANATOMY OF BACTERIOPHAGE

• SOME OF THEM HAVE BEEN NAMED AS MEMBERS OF A T
  SERIES LIKE T1,T2,T3 THE T3 AND T7 SERIERS ARE
  ICOSAHEDRAL (20 SIDED POLYGON) AND HAVE SHORT
  TAILS. THE T2,T4,T6 PHAGES HAVE AN ICOSAHEDRAL
  HEAD, A CAPSID THAT CONSISTS PRIMARILY OF 3
  PROTEINS, A CONNECTING NECK WITH A COLLAR AND
  LONG WHISKERS, A LONG TAIL, AND A COMPLEX BASE
  PLATE.

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HOW BACTERIOPHAGE WORKS

• DURING THE PROCESS OF BACTERIAL INFECTIONS BY T4
  PHAGE, AT LEAST ONE OF THE TAIL FIBERS CONTACTS
  THE LIPOPROTIENS OF THE HOST BACTERIAL CELL WALL.
• THE TAIL CONTRACTS, AND THE TAIL TUBE PASSES
  THROUGH AN OPENING THAT APPEARS IN THE BASE PLATE
  PIERICING THE BACTERIAL CELL WALL.
• THE CONTENTS OF THE HEAD, MOSTLY DNA, ARE THEN
  INJECTED INTO THE HOST CYTOPLASM

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WHEN A VIRUS KILLS, THE LYTIC CYCLE

• THE BACTERIOPHAGE EXISTS AS VIRAL DNA FREE IN
  THE BACTERIAL HOST CELLS CYTOPLASM THE VIRAL DNA
  DIRECTS THE PRODUCTION OF NEW VIRAL PARTICALS BY
  THE HOST CELL UNTIL THE VIRUS KILLS THE CELL BY
  LYSIS.

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LYSOGENIC CYCLE

• THE BACTERIOPHAGE DNA IS INTERGRATED INTO THE
  LARGE, CIRCULAR DNA MOLECULE OF THE HOST
  BACTERIUM AND IS REPRODUCED ALONG WITH THE HOST
  DNA AS THE BACTERIUM REPLICATES. IT MAY CONTINUE
  TO REPLICATE AND PRODUCE LYSOGENIC BACTERIA OR
  ENTER THE LYTIC CYCLE.

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THE PHAGE PHOLKS

• STUDENTS
• Bethany Price
• Cindy Terlizzi
• Scott Woods
• Faculty sponsor Nichol Dolby, PhD
• Faculty assistant Sherry King, PhD

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OUR CENIZA EXPERIENCE

• NOW THAT YOU KNOW WHAT BACTERIOPHAGES ARE WE
  WILL TELL YOU WHAT OUR GROUP DID WHILE HERE AT
  CENIZA
• WE THE PHAGE PHOLKS COLLECTED WATER SPECIMENS AND
  SOIL SAMPLES FROM THE LAKE, POND AND SPRING

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COLLECTING SITES
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COLLECTION SITES
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COLLECTION SITES
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COLLECTION SITES
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SAMPLES

• AFTER COLLECTING THE WATER AND SOIL SAMPLES WE
  THEN PLACED THEM INTO 15 ML TUBES WITH THE
  SPECIMEN NUMBERS ON IT
• IN THE SOIL SPECIMENS WE ADDED 10 ML OF STERILE
  WATER, MIXED THEM, AND THEN LET THEM SETTLE

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Sample Preparation

• WE THEN ATTACHED A FILTER TO A SYRINGE AND POURED
  THE WATER AND SOIL SAMPLES INTO THE SYRINGES
  USING ASEPTIC TECHNIQUE
• STEADY PRESSURE WAS APPLIED TO PUSH THE LIQUID
  THROUGH THE FILTER INTO A CLEAN RECEIVING TUBE
  MARKED WITH CORRESPONDING NUMBERS

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PETRI DISHES

• WE THEN TOOK PETRI DISHES INOCULATED WITH
  BACTERIA (LAWN EVENLY SPREAD) CONSISTING OF
  BACILLUS SUBTILIS, ESCHERICHIA COLI, AND
  PSUEDOMONAS AERUGINOSA

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Media

• EACH PETRI DISH WAS DIVIDED INTO FOUR SECTIONS
  WITH 10 ML OF EACH SAMPLE FLUID USING A PIPETOR

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INOCULATION

• WHEN ALL THE DISHES WERE INOCULATED AND THE
  LIQUID ABSORBED, WE INVERTED THE PLATES AND
  INCUBATED THEM, PLACING THEM UPSIDE DOWN AT A
  WARM TEMPERATURE ON TOP OF THE REFRIGERATOR
  ALLOWING FOR QUICKER GROWTH

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RESULTS

• MUD FROM BELOW ROCK WALL
• NO PHAGE
• WATER FROM STAGNANT POND BELOW ROCK WALL
• POSSIBLE PSEUDOMONAS PHAGE

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RESULTS

• POND EAST OF SPRING
• NO PHAGE
• MUD SAMPLE FROM SAME POND
• NO PHAGE

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RESULTS

• MOSS FROM ROCK WALL OF POND
• POSSIBLE E.COLI, PSEUDOMONAS PHAGE
• SOIL UNDER JUNIPER IN KINNEY FIELD
• POSSIBLE BACILLUS, E. COLI PHAGE

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RESULTS

• WATER SAMPLE UNDER CULVERT ANGEL BAY ENTRANCE
• RESULTS E COLI, B SUBTILIS PHAGE PRESENT

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RESULTS

• DRY SOIL FROM DRY STREAM BED NEAR DRY FALLS
• B. SUBTILUS, CLOUDY PLAQUE E.COLI PHAGE PRESENT

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RESULTS

• SOIL SAMPLE FROM BELOW DAISY PLANT ON SLOPE
• E.COLI PHAGE PRESENT

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RESULTS

• BARK FROM JUNIPER STUMP
• E.COLI PHAGE PRESENT
• LAKE TRAVIS WATER SAMPLE
• B. SUBTILIS, CLOUDY PLAQUE, E.COLI PHAGE

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RESULTS

• BIOFILM FROM LAKE SHORE ROCK
• E. COLI PHAGE PRESENT
• MATERIAL FROM UNDER ROCK
• E. COLI PHAGE PRESENT

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RESULTS

• SLIME FROM UNDER LAKE ROCK
• E.COLI PHAGE PRESENT
• ALGAE MAT FROM OFF SHORE
• E.COLI PHAGE PRESENT

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RESULTS

• SOIL AT BASE OF SHRUB ON LAKE SHORE
• E.COLI PHAGE PRESENT

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RESULTS

• SOIL FROM HIGH WATER LINE
• E.COLI PHAGE PRESENT
• SOIL FROM HIGH WATER LINE FROM BELOW SURFACE
• B. SUBTILIS, E.COLI
• POSSIBLE PHAGE

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RESULTS

• CRUMBLY ORGANIC DEBRIS FROM UNDER ROCK
• E.COLI PHAGE PRESENT
• DEEP SOIL SAMPLE FROM UNDER ROCK SANDY SOIL
• E.COLI PHAGE PRESENT

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CONCLUSION

• The most abundant bacteriophage were E.coli
  phage. Lake Travis has LOTS, implying lots of
  E.coli is present in the lake.
• We also found a few B. subtilis phage, and only a
  couple possibilities for Pseudomonas aeruginosa