Table 1: AIEC bacteria response to combination therapy
The data from replicate sets 1 and 2, represented by tables 2 and 3 respectively, has slight numerical deviations from the original set. The replicate sets also retain the general trend of having fewer colonies when strong antibiotics are used singularly and having no AIEC colonies when two strong antibiotics are simultaneously used.
Methods:   The first step that I took in order to test the response of the AIEC LF82 strain of bacteria to combination therapy that I used was to pipette 50 ul of AIEC competent cells into an eppendorf tube and place it in a tray of ice for half an hour.  Afterwards, I heat shocked the cells in the eppendorf tube in a 42 °C water bath for 60 seconds to ensure that the plasmid would enter the bateria.  Next, I left the eppendorf tube with the AIEC on ice again for 3 minutes. I then added 900 ul of LB at room temperature to the eppendorf tube and left the cells in our lab’s incubator for an hour at 37 °C  to ensure proper growth.  Afterwards, I plated 100 ul of the AIEC bacteria onto several different lb agar plates with Penicillin-Streptomycin, Kanamycin-Chloramphenicol, antimicrobial peptide (AMP), Kanamycin, SPE phase tetracycline antibiotic solutions depending on the plate and left them to incubate at 37 °C overnight for approximately 24 hours.  Plate #1 was made with lb agar and Penicillin-Streptomycin antibiotic solution, plate #2 was made with lb agar and a Kanamycin-Chloramphenicol antibiotic solution, plate #3 was made with lb agar mixed with an antimicrobial peptide (AMP) antibiotic solution, plate #4 was made with an lb agar and a Kanamycin antibiotic solution, plate #5 was made with lb agar and a SPE phase tetracycline antibiotic solution and plate #6 with only LB broth.     
The experimental groups were plate #1, which had both Penicillin and Streptomycin antibiotics used simultaneous to treat the AIEC bacterial samples, and plate #2 both Kanamycin and Chloramphenicol antibiotics used simultaneously to treat the AIEC bacterial samples.  However, there were several controls for the experiment with the positive control groups for this set being the LB+AMP, LB+Kan and LB+Spe with no second antibiotic added. The negative control group was the LB broth agar plate with no antibiotics added whatsoever. A day after observing the results of the first set of plates, sealing the original agar plates with parafilm and preserving them in our laboratory freezer, I then made one set of replicates to make sure that the trends observed from the plates were accurate.  I once again pipetted 50 ul of AIEC competent cells into an eppendorf tube and place it in a tray of ice for half an hour.  Afterwards, I once again heat shocked the cells in the eppendorf tube in a 42 °C water bath for 60 seconds to ensure that the plasmid (1) would enter the bacteria.  Next, I left the eppendorf tube with the AIEC on ice again for 3 minutes once more. I then added 900 ul of LB at room temperature to the eppendorf tube and incubated the cells for an hour at 37 °C again.  
I then plated 100 ul of the AIEC bacteria onto several different lb agar plates with Penicillin-Streptomycin, Kanamycin-Chloramphenicol, antimicrobial peptide (AMP), Kanamycin, SPE phase tetracycline antibiotic solutions depending on the plate and left them to incubate at 37 °C overnight again with plates 1-6 corresponding to the same type of antibiotic used in the original setup.  The next day I, after observing the results, I then sealed the replicates with parafilm and placed the replicate plates in the laboratory fridge. Afterwards, I made a second set of replicate plates via the aforementioned steps and preserved those plates as well to compare against the first replicate and the original plate to test for accuracy. Regarding testing the effectiveness and response of the of the AIEC LF82 strain of bacteria to sequential therapy,  the first step that I took in order to test the response of the AIEC LF82 strain of bacteria to sequential therapy that I used was to pipette 50 ul of AIEC competent cells into an eppendorf tube and place it in a tray of ice for half an hour.  Afterwards, I heat shocked the cells in the eppendorf tube in a 42 °C water bath for 60 seconds to ensure that the plasmid (1) would enter the bateria.  Next, I left the eppendorf tube with the AIEC on ice again for 3 minutes to ensure that the samples would not be overheated and to preserve the samples. I then added 900 ul of LB at room temperature to the eppendorf tube and incubated the cells for an hour at 37°C. 
 I then made two different sets of treatment groups with 7 different plates, set a and set b.  For set a, first the bacterial colonies were grown on LB broth plates mixed with weak antibiotics that after three days were then transferred to LB broth plates mixed with strong antibiotics.   I then spread and plated 150 ul of the AIEC bacteria colonies that were separated by quadrants onto several different lb agar plates with LB + Spe then Kan, LB+ Spe then AMP,  LB+ Spe then Kan/Cam, LB + Spe then P/S,  LB + Kan then P/S, LB+ AMP and P/S and then lastly the control group with no added antibiotics whatsoever.  Plates 1-4 were made with lb agar and a SPE phase tetracycline antibiotic solution.  Plate #5 was made with an lb agar and a Kanamycin antibiotic solution. Then plate #6 was made with an lb agar and an antimicrobial peptide antibiotic solution that later had Penicillin-Streptomycin added to the bacteria colonies.  Lastly, plate #7 served as a control respectively.  Two replicates sets of the original set a sample using the previously aforementioned methods to test for consistency were made. The replicates as well as the original plates were sealed with parafilm and put into our laboratory freezer.
Afterwards, the colonies grown on plates 1-6 were transferred to new LB plates with strong antibiotics incorporated into the LB agar rather than weak antibiotics. The AIEC colonies from plate #1 were also then picked and spread to an LB  plate with Kanamycin mixed in the agarand were separated by quadrants once again. An LB agar plate with antimicrobial peptide (AMP) solution was used to regrow the colonies picked from plate #2 and were spread and separated by quadrants.  The colonies from plate #3, the colonies were picked and spread on a new plate with Kanamycin-Chloramphenicol mixed in the agar in the solution and were spread and separated by quadrants once again. The colonies from plates 4-6 were spread and picked on new LB agar plates with Penicillin-Streptomycin mixed in the solution and were also spread and separated by quadrants once again.  The colonies were incubated for 24 hours to stimulate growth and afterwards were sealed with parafilm and put into our laboratory freezer.  
For set b, where a strong antibiotic was first used and mixed with the LB broth of the plates, then a weak antibiotic after a 3 day period I plated 150 ul of the AIEC bacteria colonies that were separated by quadrants onto several different lb agar plates with LB+Kan then Spe, LB+AMP the Spe, LB+Kan/Cam then Spe, LB+ P/S then Spe,  LB + P/S + Kan, LB+ P/S + AMP and then lastly the control with no added antibiotics whatsoever.   Plate #1 was made with lb agar and a Kanamycin antibiotic solution. Afterwards, plate #2 was made with lb agar and an antimicrobial peptide (AMP)  antibiotic solution. Plate #3 was made with lb agar and a Kanamycin-Chloramphenicol antibiotic solution. Then plate #4 was made with lb agar and a  Penicillin-Streptomycin antibiotic solution. Next, plate #5 was made with an lb agar and a Penicillin-Streptomycin antibiotic solution. Plate #6 was made with an lb agar and a Penicillin-Streptomycin antibiotic solution.  Lastly, plate #7 served as a control respectively.  Two replicates sets of the original set b sample using the previously aforementioned methods to test for consistency were made. Once again, replicates as well as the original plates were sealed with parafilm and put into our laboratory freezer.  
Afterwards, the colonies grown on plates 1-6 were transferred to new LB plates with weak antibiotics incorporated into the LB agar rather than strong antibiotics. The AIEC colonies from plate 1-4 were then picked and spread to an LB  plate with SPE phase tetracycline antibiotic solution mixed in the agar and were separated by quadrants. AIEC colonies from plate #5 were also then picked and spread to an LB plate with Kanamycin mixed in the agar and were separated by quadrants once again.   In addition, the AIEC colonies from plate #6 were also then picked and spread to an LB plate with a antimicrobial peptide (AMP) added agar and were separated by quadrants. The colonies were incubated for 24 hours to stimulate growth (5) and afterwards were sealed with parafilm and put into our laboratory freezer.  For all of the aforementioned treatments, regarding each of the plates, for every 25 ml of lb agar, 1 ml of antibiotic solution was used in conjunction with the agar so that each plate had an antibiotic concentration that was proportional to the amount of AIEC bacteria plated.
Results: I grew the AIEC LF82 bacteria on six agar plates (as previously mentioned) with Penicillin-Streptomycin, Kanamycin-Chloramphenicol, antimicrobial peptide (AMP), Kanamycin, SPE phase tetracycline antibiotics to test their resistance and noticed that all of the antibiotics alone, proved to be quite infective since the untransformed AIEC bacteria still were able to grow very similarly to how they would on a regular lb agar plate. Antibiotic and probiotic therapies appear to be a very poor potential choice for therapeutic treatment of ileal Crohn’s disease and IBD, at least at first glance.  However, I noticed that plates #1 and #2 were able to completely halt the colonization and therefore the growth of AIEC LF82 bacteria.
 
  The effects of combination  therapy using antibiotics ended up being more potent than initially predicted.  When two strong antibiotics used via combination therapy tended to actually be more successful than the weak-strong sequential therapy treatment used in set a. The effects of combination therapy on the AIEC bacterial colonies also affected the colonies quicker than expected in only a 24 hour period.  The significance of this is that clinical combination therapy using antibiotics can tend to not be the most time effective, however the P/S and Kan/Cam antibiotic solutions were able to act and work quickly on eradicating the AIEC colonies when used in conjunction. When two strong antibiotics were used in conjunction they tended to interfere with AIEC bacteria reproduction and halted the growth and colonization of AIEC colonies.