SEA Phage Paper

In the beginning of lab, we were advised to obtain a nutrient agar petri plate, which is used for the cultivation of microbes supporting growth of non-fastidious organisms. Since it contains many nutrients, a wide variety of bacteria and fungi can grow. Taking the plate,

The next step of the project included preparing a Gram stain to discover the cell shape, arrangement, and if the bacteria is gram positive or

The overall goal is to either discover a new novel phage that has not yet been discovered, or provide new research on an already discovered phage. The isolation of the phage takes several steps under the streak protocol that is present in materials and methods section of this report.

The reversion rate of either phage rII 29, or phage rII 31 was determined by the infection of permissive host E. Coli B, or with non-permissive host E. Coli K to either bacteriophages rII 29, or bacteriophage rII 31. This infection was plated using the soft agar technique. After incubation, the titer of revertants phage was determined by the number of plaques formed on E. coli K plates. The reversion rate equation was the following,

Often scientists work with bacteria that do not come in a labeled test tube— for example, bacterial samples taken from infected human tissue or from the soil—and the scientist must then identify the unknown microorganism in order to understand what behavior to expect from the organism, for example, a certain type of infection or antibiotic resistance. However, because of the relatively few forms of bacteria compared to animals and because of the lack of bacterial fossil records due to their asexually reproductive nature, the taxonomy used to classify animals cannot be applied to bacteria (Brown 275). In order to classify unknown bacteria, a variety of physiological and metabolic tests are available to narrow a sample down from the fathomless number of possibilities into a more manageable range. Once these tests have been performed, the researcher can consult Bergey’s Manual of Determinative Bacteriology, a systematically arranged and continually updated collection of all known bacteria based on their structure, metabolism, and other attributes.

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

Bacteriophages were first discovered about 100 years ago by Frederick Twort and Felix D'Herelle. Bacteriophages or commonly known as “phages” are viruses that infect bacterial hosts. Bacteriophages come in multiple shapes and sizes. But a good amount of them are tailed viruses that contain double stranded DNA. The head of the bacteriophages has a protein shell which is attached to the tail. Some phages differ as some don’t have double stranded DNA they may be single stranded. Phages are incapable of reproducing on their own. In fact, they need a bacterial host to reproduce. Like any virus, bacteriophages are very fixed to discrete hosts. The host bacteria that we are using is called arthrobacter. Arthrobacter is a genus of bacteria found in soil (Pope et al 2016). Arthrobacter can help reduce hexavalent chromium which can cause some symptoms such as irritations to humans. Hexavalent chromium may also cause lung cancer if inhaled. The SEA PHAGE project was broken down into two parts. The first part which we are currently doing is isolating, purifying and amplifying the bacteria from our soil samples that we collected in the beginning of the year.

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

Unknown lab report# 24 Introduction Microbiology is a study of organisms that surrounds us every day. It requires an educational understanding to identify organisms, and the ability to distinguish different bacteria’s. In applying the learning process of the different bacteria’s, unknown bacteria samples are given to be studied and identified.

Prokaryotes are ubiquitous, successfully adapting to diverse environments as well as developing symbiotic relationships with host organisms (Lengeler, Drews, & Schlegel, 1999). Prokaryotes may have both autotrophic and heterotrophic characteristics. A cyanobacteria is photosynthetic, commonly called blue-green algae, and may produce toxins (Crayton, 1993). Bacteria are most commonly associated in the general

The bacteria that was contained within Unknown tube #12 is believed to be Pseudomonas aeruginosa, Figure 1. The bacteria tested to be Gram Stain negative, producing a pink, red color retained from the staining process. When the species of bacteria was plated on nutrient media, the cells produced an irregular and spreading configuration as shown in Figure 2. This same plating test provided the margins and elevation, lobate and hilly, respectively. The specimen was stabbed in a Fluid Thioglycollate Medium (FTM) tube using an inoculated loop of the bacteria. The results of this experimentation indicate the type of oxygen requirement of the bacteria. The test found the bacteria to be aerobic as colonies of the bacteria began to form along the top of the FTM tube (Manual 2017).

When Dubos began to study soil, especially swamp soil, he was trying to find organisms that were able to break the protective coat surrounding human lobar pneumonia bacteria (an infection in one or both lungs, the bacteria causes the lung’s air sacs to become inflamed and filled with pus, fluid, and cellular debris.). He succeeded in isolating a swamp bacteria that produced an enzyme (Tyrothricin) which abled the human body to attack the pneumonia bacteria germ. Later, Dubos isolated other bacteria, like Bacillus brevis which can consume deadly germs. Dubos’s discovery of Tyrothricin made other discoveries of broad spectrum

These are gram positive bacteria which are found in acidic condition of pH 4 to 4.5 – acidophilic.

There are several advantages to phage therapy with target specificity being one of the main ones. Most phages will only lyse bacteria of a particular species or a specific strain meaning that the human microbiota should not be affected. In contrast, antibiotics are less specific and so treatment usually disrupts this flora, allowing opportunistic bacteria to thrive. This specificity is also a limitation as the bacteria causing the infection needs to be determined otherwise the treatment is likely to be useless. Such problems can however be addressed with the use of phage combinations like ‘cocktails’ used in the above study.

Wolbachia is a widespread obligate intracellular bacterium, and it’s a member of Rickettsiales order. Wolbachia can significantly alter the reproductive ability of its host and cause re-productive parasitism in arthropods species, including male killing, feminization, partheno-genesis and cytoplasmic incompatibility (CI). Bacteriophage WO is a kind of virus that can infect Wobachia cells and cause the host bacterial cell lysis (bursting). When WO phage at-tached to bacterial cells, the phage genome will be injected and insert into the Wolbachia chromosome, that called prophage WO. Prophage is only a latent stage, when the host bacte-rial cells are damaged by UV radiation or certain chemical substances, the prophage can be released and cause propagation of new phage by lytic cycle. However, if WO phage in wMel strain of Wolbachia can become lytic is still not known. In this study, we investigated that whether prophage WO can be reactivated by stressors (extracellular Wolbachia was treated by doxycycline or rifampicin). From our results of real-time PCR and proteomic analysis we proved that doxycycline and rifampicin treatment did not appear to generate lytic phage. Wolbachia is a widespread obligate intracellular gram-negative α-proteobacteria (Wu et al., 2004), it is a member of Rickettsiales order and belongs to the family of Anaplasmayaceae (Darby et al., 2013). Wolbachia has wide host ranges, including 47% of filaria nematodes (Ferri et al., 2011) and 40% of arthropods