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Aryal, Sagar. “Acid Fast Stain (Kinyoun-Cold Method)- Principle, Procedure and Result Interpretation.” Microbe Notes, 11 Jan. 2020, microbenotes.com/acid-fast-stain-kinyoun-cold-method-principle-procedure-and-result-interpretation/.

Cowan, M. Kelly, et al. Microbiology Fundamentals: a Clinical Approach. McGraw-Hill Education, 2019.

Crownover, Julie. “Nocardia Farcinica.” Microbewiki, 1 July 2011, microbewiki.kenyon.edu/index.php/Nocardia_farcinica.

De La Iglesia, Pedro, et al. “Fatal Pulmonary Nocardia Farcinica Infection.” Journal of Clinical Microbiology, American Society for Microbiology, Mar. 2002, www.ncbi.nlm.nih.gov/pmc/articles/PMC120238/.

Grant, Maurice, et al. “Final Diagnosis -- Infection of Nocardia Farcinica.” Final Diagnosis -- Case 315, 0AD, path.upmc.edu/cases/case315/dx.html.

Hindawi. “Fatal Nocardia Farcinica Bacteremia Diagnosed by Matrix-Assisted Laser Desorption-Ionization Time of Flight Mass Spectrometry in a Patient with Myelodysplastic Syndrome Treated with Corticosteroids.” Figure 1, 2013, www.hindawi.com/journals/crim/2013/368637/fig1/.

Lahiri, K K, and S K Chatterjee. “A SIMPLE COLD STAINING METHOD FOR ACID FAST BACILLI.” Medical Journal, Armed Forces India, Elsevier, Oct. 1994, www.ncbi.nlm.nih.gov/pmc/articles/PMC5529720/.

“Nocardia Culture.” Fairview University Diagnostic Laboratories, 2015, www.childrensmn.org/references/Lab/microbioviral/nocardia-culture.pdf.

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What are you most looking forward to learning in microbiology?

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What are you most looking forward to learning in microbiology?

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I am most looking forward to learning more about antimicrobial susceptibility and the testing guidelines that go along with that to figure out how to narrow down specific bacteria or viruses and to treat them. I am also looking forward to learning about different bacteria types and how they can cross over in the symptom aspect and how to narrow down to what specific strain we are looking at or for and how to determine getting rid of it if the mass can be treated with antibiotics or are there other ways to treat bacterial masses.

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The patient was placed on a 6 week course of intravenous Bactrim and Imipenem. These are two different types of antibiotics that showed the minimal amount of bacterial growth. These two make the perfect cocktail to treat N. farcinica as they both were the most susceptible to further growth of the bacteria. He was further placed on oral antibiotics after having been discharged, these he will take everyday for a year as to make sure that the bacteria will not come back or grow further if the 6 week course does not decrease the bacterial load enough for the body to finish it off or to rid the body of it completely.

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How did the health care team use the results to make a diagnosis?

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How did the health care team use the results to make a diagnosis?

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Based on the growth, stains, and results from the susceptibility testing they were able to narrow it down to Nocardia farcinica as the growth pattern was consistent with Nocardia spores and the susceptibility testing helped to narrow further down to the strain that it was. Since the patient also had suspected lung cancer which they then thought the mass was actually N. farcinica showed that the patient already had a suppressed immune system due to age so it was easy for the bacteria to enter the body and spread like a wildfire.

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Antimicrobial testing is another microbiology test that was ordered in the case study. Within this as they narrow down the bacteria they must have an antibiotic which will help to kill off the rest of the bacteria and to prevent the growth of further bacteria. In this case bactrim and imipenem helped to prevent the most growth of bacteria and showed the most susceptibility. Antibiotic testing is important as you need to be able to recognize antibiotic resistant bacteria such as MRSA.

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Stains are utilized to to identify gram-positive and gram-negative bacteria as well as to identify their shape and arrangment as it can help to narrow down the strains of bacteria and what type of bacteria they are.

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Cultures are used to study the growth of bacteria and the patterns that show from this growth. After the stains were done they had narrowed the bacteria down to Nocardia spores and from there they set up two different types of culture growths. Quad plates and chocolate plates were used to further help the Nocardia spores to grow and assist the microbiologists in helping to identify what strain they are dealing with.

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Two sets of cultures were ordered as well as two types of stains. There was also a set of tests ordered to test the bacteria's resilience.

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What are the patient's signs and symptoms?

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What are the patient's signs and symptoms?

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The patient exhibited and reported symptoms of dizziness, headaches, and gait instability that continued during a three week period. The patient has a suspected lung cancer as well but did not note any symptoms they were experiencing respiratory wise.

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Kinyoun stains are similar to gram stains in certain aspects. They are performed at room temperature, need the slide to be heat fixed. The preparation before the slide can be dyed are the same as acid-fast and gram stains. After you have prepared the slide you will want to place it on the rubber bands bacteria facing up, over the container. You begin by placing carbol fuschin over the bacteria side of the slide for 3 to 5 minutes at room temperature allowing for the bacteria to absorb the dye. Just as with the others you will wash the slide with distilled water to rid of excess dye. Then using a 1% sulfuric acid you will decolorize the slide similar to the acid fast stain. You want to run the decolorizer over the slide until the run off is clear or after about 3 minutes then rinse again using distilled water to clear off the acid. You then place methylene blue over the slide for 60 seconds then rinse with distlled water again blotting the edges dry then letting it air dry.

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An acid-fast stain is done over boiling water generally half a beaker full, in class we used 500mL beakers so you would fill it up to the 250mL mark then let it come to a steady boil before sliding the rack over it. When you have successfully placed your rack to keep the slide from falling in you will want to place your heat fixed slide on top the rack covered with the bibulous paper from the tray, tearing a piece that is big enough for the slide but not overly big that it is hanging off. Then cover the paper with carbol fuscion you will continue to add this every few seconds as it will dry out fairly quickly and this will go on for about 5 minutes over the boiling water. After the five minutes have passed remove the paper and throw it in your container and put the slide on top of your rubber bands. Using distilled water wash the excess dye off then using the acid alcohol you will rinse the slide until the run off is clear. Then using malachite green you will let that sit on the slide for around 1 minute or longer, then proceed to rinse with distilled water then blot dry the edges and let the slide dry.

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Gram stains do not require the use of boiling water to help absorb the dye. With gram stains you use crystal violet after heat fixing the slide to help dye the gram positive bacteria violet. You let the crystal violet sit on your slide for around 60 seconds before using distilled water to rinse the excess dye off. You then utilize iodine to help seal the color with gram positive bacteria again waiting a full minute before rinsing with distilled water. Then to decolorize the gram negative bacteria you use the alcohol with the slide on a slant over a container you begin to pour it over till the run off is no longer purple. You proceed to use the safarin to dye the gram negative bacteria to red, leaving this on again fro a full minute then rinsing with distilled water and blotting the edges of the slide dry but not the side with bacteria.

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What staining techniques were used to characterize the bacterium in the case study?

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What staining techniques were used to characterize the bacterium in the case study?

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The staining techniques used in the case study consisted of Gram Staining, Acid-Fast Staining, and Kinyoun Staining. Gram stains and acid-fast stains we produced in class with multiple different bacteria. With these stains we used the dyes crystal violet, safarin, and carbol fuschion. These stains had different yet similar techniques. To start all three you will use sterile technique and asepsis to place the bacteria on the slide and maneuvering it to spread thinly with the drop of distilled water over the slide to dry faster. You then will proceed to heat fix the slides before moving to the dye steps.

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What is the typical shape and arrangement? Are there any unique features that can be seen under a light microscope?

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What is the typical shape and arrangement? Are there any unique features that can be seen under a light microscope?

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Typical arrangement and shape are branch like structures and bacillus shaped bacteria. The bacteria is arranged in a streptobaccilus pattern though with the projections of others off the sides I'm not sure that this would still be the correct classification of arrangement. Unique features that N. farcinica possesses looks like filaments projecting off the bacteria, though this could just be from the picture being blurry when zoomed in.

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What does N. farcinica look like under a light microscope?

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What does N. farcinica look like under a light microscope?

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Under light microscopy N. farcinica appears to be crystal violet as it is a weak gram-positive bacteria. Looking at a slide found on a medical journal (below) it appears in rod shaped structures that can be described as streptobacillus. They appear as small branch like arrangements similar to that of a tree branch or vine.