Legionella Pneumophila

How do you grow it?

• 1. What is the BSL level for your microbe?

Legionella pneumophila has a BSL level of 2.

• 2. What type of agar do you need?

Legionella pneumophila will need Buffered Charcoal Yeast Extract agar.

• 3. Does it need to be in aerobic/anaerobic conditions?

Legionella pneumophila needs to be in aerobic conditions to grow. The atmospheric conditions for growth is 5% CO₂.

• 4. What is the procedure for growing it once it arrives?

The procedures for growing my microbe are:

1. Open the vial according to the enclosed instructions on the ATCC website.

2. Rehydrate the entire pellet with about 0.5 mL of CYE Buffered Broth. Aseptically transfer the contents to a 5-6 mL tube of CYE Buffered Broth. In addition, extra test tubes can be inoculated by transferring 0.5 mL of the primary broth tube to these secondary broth tubes.

3. Use many drops of the primary broth tube to inoculate a CYE Buffered plate or agar slant.

4. Incubate at 37˚C for 48-72 hours in 5% CO₂.

• 5. Are there any other special instructions?

It is important to pay attention to the quality of media used as Legionella species are nutritionally fastidious and inhibited by poor quality media. In addition, it is very essential to check the pH of the medium when cool. Moreover, unbuffered medium should not be used and if the medium is exposed to light, it may lead an increase of peroxides that inhibit bacterial growth.Also, before rehydration, the vials can be stored +2 to +8˚C for long periods of time with minimal loss of viability. Furthermore, subculturing to maintain culture is not encouraged due contamination and chances of developing mutations. It is best to harvest the cells and store them at -70˚C or below to minimize changes.

• 6. When was the first documentation of it? Who discovered it?

The first documentation of my microbe is in 1979 and it was discovered by Paul H. Edelstein.

1. Is it a pathogen? What disease does it cause? Or does it have any benefits to the host? How can infection or contraction of the disease be prevented?

Legionella pneumophilais a pathogen that causes Legionnaires’ Disease, also called legionellosis. There is no vaccine for preventing Legionnaires’ Disease. Instead, in order to prevent the contraction of this disease, it is important that managers maintain our man-made water systems such as plumbing systems and cooling towers in order to reduce the risk of Legionella pneumophilagrowth. In addition, to help prevent contracting Legionnaires’ Disease at home, it is important to remove and clean all shower heads every 3 months as they are a common place where Legionella pneumophilacan grow.

2. Where is it traditionally found?

Legionella pneumophilais traditionally found in soil and fresh water in lakes, rivers, and hot springs. In societal settings, it can grow if water is not treated properly. Some common sources of where the bacteria can be found is in water used for showering, cooling towers, fountains, hot tubs, and plumbing systems.

3. Why do people care about it? Is there a commercial or ecological application, other oddities about your adopted microbe?

The reason people care about Legionella pneumophilais because it is readily found in sources of water and from everyday sources such as tubs and showers. For instance, in 2014, the Flint, Michigan Water Crisis erupted as a result of switching the drinking water supply from to the Flint River. As a result, many individuals got Legionnaire’s disease due to the contaminated water and the fact that the pipes that the water flowed through was old and leached lead. Ever since this crisis, people have been more aware aboutLegionella pneumophilaand thus, the applications this bacteria has on our daily lives is seen through the preservation and maintenance of these man-made water systems.

4. What Antibiotics work against your bacteria? Are there any other treatments for it?

Many antibiotics do in fact work against Legionella pneumophila.If an individual has Legionnaires’ disease, they require treatment with antibiotics and after becoming sick, they typically need care in the hospital in order to successfully recover. The two most effective classes of antibiotics are the marcolides and the quinolones. Treatments such as azithryomycin and levofloxacin work effectively. However, there are also other antibiotic agents such as tetracycline and doxycycline that have been proven to be effective. In addition, in more severe cases of Legionnaires’ disease, adjunctive glucocorticoids and extracorporeal membrane oxygenation may also be used.

The primers I found for Legionella pneumophila were:

Forward

· 5'-AGGGTTGATAGGTTAAGAGC-3'

· Melting Temperature: 49.7 ˚C

· GC Content: 45%

Reverse

· 5'-CCAACAGCTAGTTGACATCG-3'

· Melting Temperature: 51.8˚C

· GC Content: 50%

The specific 16s sequence (in FASTA format) for Legionella pneumophila is:

>LC486880.1 Legionella pneumophila Niigata 1675 gene for 16S ribosomal RNA, partial sequence

TGAAGAGTTTGATCCTGGCTCAGATTGAACGCTGGCGGCATGCTTAACACATGCAAGTCGAACGGCAGCATTGTCTAGCTTGCTAGACAGATGGCGAGTGGCGAACGGGTGAGTAACGCGTAGGAATATGCCTTGAAGAGGGGGACAACTTGGGGAAACTCAAGCTAATACCGCATAATGTCTGAGGACGAAAGCTGGGGACCTTCGGGCCTGGCGCTTTAAGATTAGCCTGCGTCCGATTAGCTAGTTGGTGGGGTAAGGGCCTACCAAGGCGACGATCGGTAGCTGGTCTGAGAGGATGACCAGCCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGGGCAACCCTGATCCAGCAATGCCGCGTGTGTGAAGAAGGCCTGAGGGTTGTAAAGCACTTTCAGTGGGGAGGAGGGTTGATAGGTTAAGAGCTAATTAACTGGACGTTACCCACAGAAGAAGCACCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGGTGCGAGCGTTAATCGGAATTACTGGGCGTAAAGGGTGCGTAGGTGGTTGATTAAGTTATCTGTGAAATTCCTGGGCTTAACCTGGGACGGTCAGATAATACTGGTTGACTCGAGTATGGGAGAGGGTAGTGGAATTTCCGGTGTAGCGGTGAAATGCGTAGAGATCGGAAGGAACACCAGTGGCGAAGGCGGCTACCTGGCCTAATACTGACACTGAGGCACGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCTGTAAACGATGTCAACTAGCTGTTGGTTATATGAAAATAATTAGTGGCGCAGCAAACGCGATAAGTTGACCGCCTGGGGAGTACGGTCGCAAGATTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGATGCAACGCGAAGAACCTTACCTACCCTTGACATACAGTGAATTTTGCAGAGATGCATTAGTGCCTTCGGGAACACTGATACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGTAACGAGCGCAACCCTTATCCTTAGTTGCCAGCATGTGATGGTGGGGACTCTAAGGAGACTGCCGGTGACAAACCGGAGGAAGGCGGGGATGACGTCAAGTCATCATGGCCCTTACGGGTAGGGCTACACACGTGCTACAATGGCCGATACAGAGGGCGGCGAAGGGGCGACCTGGAGCAAATCCTTAAAAGTCGGTCGTAGTCCGGATTGGAGTCTGCAACTCGACTCCATGAAGTCGGAATCGCTAGTAATCGCGAATCAGCATGTCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTGCACCAGAAGTAGATAGTCTAACCTTCGGGGGGACGTTTACCACGGTGTGGTTCATGACTGGGGTGAAGTCGTAACAAGGTAGCCGTAGGGGAACCTGCGGCTGGATCACCTCCTT