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Future Learning Aspirations in Microbiology

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Future Learning Aspirations in Microbiology

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I am interested to learn more about the effects of bacteria on animals vs. humans, especially when that bacterium is originated from animals and transferred to humans. Tularemia is not the only case in which animals can transfer that bacterium, and with present day situations like that of COVID-19, I wish to learn more about how this occurs, and if these differing species have the same affects.

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Before the lab results confirmed the patient's diagnosis as Tularemia, the physician was strongly under the impression that the granulomatous abscess was a result of LGV. This resulted in the prescription of doxycycline, which is used to treat a variety of bacterial infections. This antibiotic stops bacterial growth and treats inflammatory conditions (Puckey 2019). While doxycycline is usually not the primary antibiotic chosen to treat Tularemia, it can be effective in some cases. Because of this, the physician chose to continue treating the patient with doxycycline even after learning the correct diagnosis. Futher inquiry led to the discovery of exposure. The patients had encountered a dying baby rabbit and came into contact with it. This is believed to be the cause of the patient's condition. The patient went on to have a full recovery.

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Swollen lymph nodes of an individual with glandular Tularemia. (Credit: Zahid 2018)

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Caused by F. tularensis which is transferred during animal (rabbits, squirrels, and birds) and tick exposures.

Symptoms (Feldman 2003):
- Chills
- Headache
- Malaise
- Forms of Manifestations including ulceroglandular (most common form with ulcer developments at cutaneous or mucous membranes) and glandular (ulcers not present).

Treatments (Feldman 2003):
- An Aminoglycoside, usually Streptomycin (primary choice).
- Gentamicin

Comparison to patient's symptoms:
The patient experienced granulomatous abscesses which corresponds with the glandular form of Tularemia.

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F. tularensis Gram Stain (Credit: Dr. W.A. Clark, CDC. PHIL)

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Cause of tularemia, sometimes referred to as rabbit fever.

Biochemical Characteristics:
- Coccobacillus
- Nonmotile
- Aerobic
- Gram negative
- Oxidase negative
- β-lactamase positive
- Grows well on chocolate agar

Similarities to bacteria extrapolated from patient: The bacteria from the patient grew Gram-negative bacillus on a chocolate agar plate, which matches up to the biochemical characteristics of F. tularensis.

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- On a chocolate agar plate, small rods were present. The bacteria were Gram stained. This staining technique is used to determine cell wall structure which can help differentiate bacteria. The bacterial growth was Gram-negative.

- Pyrosequencing showed an 85% consistency to Francisella tularensis.

- The PCR reaction confirmed the organism to be Francisella tularensis.

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Scidmore M. 2005. Cultivation and Laboratory Maintenance of Chlamydia trachomatis. Current protocols in microbiology. https://doi.org/10.1002/9780471729259.mc11a01s00. https://pubmed.ncbi.nlm.nih.gov/18770550/.

008565: Chlamydia trachomatis Culture | Labcorp. wwwlabcorpcom. https://www.labcorp.com/tests/008565/i-chlamydia-trachomatis-i-culture.

CDC. Details - Public Health Image Library(PHIL). philcdcgov. [accessed 2023 Feb 10]. https://phil.cdc.gov/details_linked.aspx?pid=1904.

Feldman KA. 2003. Tularemia. Journal of the American Veterinary Medical Association. 222(6):725–730. doi:https://doi.org/10.2460/javma.2003.222.725. [accessed 2020 Nov 24]. https://pubmed.ncbi.nlm.nih.gov/12675294/.

Lorenz TC. 2012. Polymerase Chain Reaction: Basic Protocol Plus Troubleshooting and Optimization Strategies. Journal of Visualized Experiments. 63(63). doi:https://doi.org/10.3791/3998.

Medico Sage. 2021 Nov 3. BACTERIAL CULTURE MEDIA – Microbiology. medicosagecom. [accessed 2023 Feb 10]. https://medicosage.com/bacterial-culture-media/#:~:text=A%20culture%20medium%20is%20a%20solid%20or%20liquid.

Melisa Puckey. 2019. Doxycycline. Drugscom. https://www.drugs.com/doxycycline.html.

Zahid H. 2018 Sep 8. An interesting case of Glandular Tularemia. specialitymedicaldialoguesin. [accessed 2023 Feb 10]. https://speciality.medicaldialogues.in/an-interesting-case-of-glandular-tularemia.

Zhao F, Bapat B. 2016. The Role of Methylation-Specific PCR and Associated Techniques in Clinical Diagnostics. In: Epigenetic Biomarkers and Diagnostics. Academic Press. https://www.sciencedirect.com/science/article/pii/B9780128018996000085.

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A 50-year-old married minister with swollen lymph nodes in his groin area (Inguinal Lymphadenopathy). Both husband and wife claim they have had no other sexual encounters.

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The physican took a lymph node biopsy, which gave results of a granulomatous abscess. Physican believed this abscess to be linked to lymphogranuloma venereum (LGV), which is caused by a sexually transmitted infection called Chlamydia trachomatis. When both spouses continued to refuse having had other sexual encounters outside of their marriage, other tests were run to determine cause.

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A tissue specimen was sent to a laboratory for culture testing.

Cultures Used:
Chlamydia culture - Swab epithelial cells are grown on a tissue culture system and Chlamydia trachomatis is detected by fluorescent antibody staining (Scidmore 2005; Labcorp). Transportation survival is low, so these cultures are not always accurate (Labcorp). In the case of the patient, the culture was negative for Chlamydia trachomatous growth, but because of a transportation mistake, the physician chose not to rule out Chlamydia completely.

Bacterial culture - The epithelial cells are grown on/in a solid and/or liquid mixture of nutrients. These cultures are used for a variety of tests to distinguish and identify bacteria and promote growth (Medico Sage 2021).

Other Tests:
Pyrosequencing - This is a DNA sequencing technique that uses the release of pyrophosphate during the addition of nucleotides and its corresponding light signals to identify organisms (Zhao and Bapat 2016).

Polymerase Chain Reaction (PCR): This technique amplifies segments of DNA for a broader analysis of DNA templates of unknown organisms (Lorenz 2012).