In order to isolate bacteriophage specific for Salmonella, we will following procedures below.

Some phages infect bacteria to destroy them, which is called lysic lifestyle, while other phages infect bacteria and stay dormant inside them for a while, which is referred to as a lysogenic lifestyle. A phage infects the bacteria cell by injecting its genetic material into the bacteria’s cytoplasm. This allows the bacteria synthesis process to start making the phage’s genetic code instead of its own. Once the bacteria have made enough phages to handle, the walls will break and release all of the phage that was created. The phages that were made are now resetting the process and beginning again by infecting the other near bacteria by injecting their genetic material once again. Those phages that stay within the bacteria and not burst the bacteria will continue to reproduce the phages own genetic code. (Griffiths,

5 drops of chloroform were added to the mixture and incubated at room temperature for 15 minutes to kill, and lyse the bacteria cells which allows all the unadsorbed phages to be accounted for when determining the titer of unadsorbed phage with susceptible bacteria, E. Coli B. (Biology Dept.). 0.1 ml of E.coli B was added to the 10 fold dilution. Using soft agar technique, the unadsorbed phage were plated. After incubation, the titer of unadsorbed phage was determined by counting the number of plaques on the plate from the infection of single E. coli cells in the assay tube by a free phage particle before

6 petri dishes were labeled, 3 petri dishes P 105 B, P 106 B, P 107 B, and the other 3 petri dishes P 10 K, P 102 K, P 103 K and they were set aside. A series of dilutions for the 1x bacteriophage T4 rII

This lab experiment was done for the purpose of learning how to determine a gram negative bacterium based on multiple tests learned throughout the semester. My gram negative unknown bacterium given to me was Salmonella typhimurium based off of the following tests; Triple Sugar Iron Agar (TSIA), Sulfate Indole Motility (SIM), Methyl Red (MR), Voges-Proskaur (VP), Citrate, Urea Hydrolysis, and Gelatin Hydrolysis. Each test performed gives results such as motility, acid production, fermentation, carbon requirements, or detection of certain coenzymes. With a process of elimination, I determined which bacteria it was not and which bacterium I had, S. typhimurium. The expectation was to master the techniques for each test and utilize the results to determine the unknown bacterium I was given within a two-week period.

70µL of competent E.coli are added to both test tubes; pUC18 and Lux (Alberte et al., 2012). Both test tubes are then tapped and placed back into the ice bath for 15 minutes. While waiting, another test tube is obtained, filled with 35µL of competent cells and labeled NP for no plasmid. A water bath is preheated to 37 degrees Celsius and all three labeled test tubes are inserted into the bath for five minutes (Alberte et al., 2012). Using a sterile pipet 300µL of nutrient broth are inserted into both the control and Lux test tubes and 150µL are inserted to the no plasmid test tube to increase bacterial growth. All three test tubes are then incubated at 37 degrees for 45 minutes. Six agar plates are obtained and labeled to correspond each test tube, three of the plates contain ampicillin. A pipet is used to remove 130µl from each test tube containing a plasmid and insert it into the corresponding agar plate. For this, a cell spreader is first

The main objectives of this experiment included making dilutions of solutions, plating phage or bacteria, and determining the number of bacterial viruses or phage in a suspension. It was also conducted to demonstrate that two different mutants of phage T4 can exchange genetic material to give rise to wild-type phage. The experiment was used to distinguish mutants from wild-type by their host specificity. The recombination in bacteriophage was performed to determine the concentration of unadsorbed phage from the U series plates, total concentration from B series, and concentration of

There were two tubes used in this process: the tube that contained the primary culture and the tube that contained the nutrient agar where the unknown bacteria would grow. First, the inoculating loop was flamed. After removing the caps of both the test tubes, they were flamed to prevent contamination of the unknown bacteria. The inoculating loop was cooled for a few seconds and was then placed into the test tube containing the bacteria. The inoculating loop with the bacteria was placed into the nutrient agar test tube for cultivation. Before the test tubes were capped, they were flamed once again. Also, isolation of the unknown bacteria had to completed. Nutrient agar was placed in the petri dish, and was left to gel for a few minutes. After the agar gelled, the inoculating loop was used to acquire bacteria and streak the unknown onto the plate for

This project is all about isolating bacteriophage in soil. They come in different sizes and shapes, each to their own unique look. Phages have a protective protein head that contains DNA and a hollow tube tails (http://phages.org/bacteriophage/). Since bacteriophage cannot reproduce and replicate themselves, they need a host to do the work for

You can grow microorganisms in liquid media or on solid media. Most bacteria can be grown in labs as long as the media contain a source of the major nutrients; carbon, nitrogen, sulphur, and phosphorous. They may also need to have other nutrients. These nutrients are made into a broth and the pH and salinity can be adjusted. The nutrient broth is placed in test tubes, which are plugged with cotton wool, capped with foil and then sterilised in an autoclave, at 121c for 20minutes. These tubes are then cooled before they are inoculated. To prepare the streak plates we dipped an inoculating loop into ethanol then placed it in the flame until the loop glowed red. Still holding the inoculating loop by its handle we removed the lid from

For the Gram stain, I used a microscope slide, wash bottle of water, clothespin, and the reagents crystal violet, Gram’s iodine, ethyl alcohol, and safranin to observe whether the organism was Gram negative or positve. The method was placing and heat fixing a loopful of the unknown organism on the slide. The organisms were then stained with crystal violet for a minute, rinsed off, flooded with iodine for a minute, rinsed off, decolorized with alcohol for 30 seconds, and then finally stained with the counter-stain, safranin, for a minute. I streaked the unknown onto a TSA plate and incubated at 35 C. With a pure colony, we performed a second gram stain procedure and inoculated it into a TSA slant. I used BCP Lactose broth and a loop to perform a BCP Lactose test. The broth was inoculated with a loopful of the unknown and incubated

Before the discovery of antibiotics, research was being conducted on the use of bacteriophages for use in treating infections.16 Among the most abundant type of organism on the planet, bacteriophages present a solution that is species-specific and has less side effects than antibiotics, since eukaryotic cells lack the phage receptors needed for a phage infection.16 Of the two types of bacteriophages, lytic and lysogenic, lytic phages are of great interest due to the fact that they replicate in and subsequently lyse the bacterial host cell.16

There are many differences between bacteriophages and bacterium. Bacteriophages (also known as phages) are viruses that are obligate intracellular parasites, therefore, need a specific host in order to for this virion to leave its dormant state. The bacterium serves as the host in which the virus inhabits, allowing for it to grow and replicate. According to the case study by Kari Mergenhagen, the virus also obtains and enormous size compared to the bacterium. It has special structures, such as, a protein coat (capsid) which is a nucleic acid genome. On the other hand, bacterium is not simply defined as being a host for phages. Bacterium is a microorganism which is unicellular, has cell walls, lacks of organelles and nucleus, and can reproduce

In order to obtain well-isolated discrete colonies, the quadrant streak and spread technique was used. This allowed dilution of the original microbial material over the entire surface of the plate. As the original sample was diluted by streaking and spreading it, over successive quadrants the number of organism decreases. Usually by the third or fourth quadrant only a few organism were transferred on the by the inoculating loop and theses produce a few isolated

A sterile pipette was used to add 0.1ml of E. coli culture to the pH 3.0 tube. This was then repeated for the tubes at pH 7.0 and pH 9.The tubes were then incubated at 37oC for 48 hours. This was then repeated for saline culture of Saccharomyces cerevisiae but incubated for 72 hours at 25oC.

Understanding the importance of these seemingly simple, yet extremely multi-variable and diverse particles can be beneficial in the coming future of the science world. Scientists Breeann Kirby and Jeremy J. Barr discuss the growing failure of antibiotics over the last few decades, and how researchers are now turning to lytic phages for bacterial control in the US food industry, commercial agriculture, and veterinary medicine (Kirby et al. 2013). Thus, proving that bacteriophages can play a major role in multiple industries that fuel society and the economy forward. Which only highlights the importance of this up and coming research field. An industry that is split upon this new form of treatment among a ray of issues in the medical community. However, this debate should not be overlooked but rather welcomed so that progress can be made. Carl Zimmer explains that it may be hard to imagine a world before antibiotics, let alone a world where antibiotics are not the only weapon with the ability to fight against bacteria (Zimmer, 2011). There is no denying that antibiotics are becoming less effective every year as the resistance to drugs continues to evolve. This is an issue that cannot be ignored which is where talk about Phage therapy begins to emerge which may ultimately prove to be of great clinical value one day. There are