My Husband's Beard

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A lab report I wrote after taking a sample from my husband's beard.

Submitted: May 17, 2016

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Submitted: May 17, 2016

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Brandi Opsal

 

May 8, 2016

Male Beard Specimen

Gamella haemolysans

 

 

 

Introduction

Bacteria are everywhere. Identification of environmental unknowns is one of the most important jobs of a microbiologist. The process of identifying environmental bacteria is significant because it enables scientists to understand where microorganisms live naturally and how they may impact human life. Many microorganisms are capable of causing infectious disease, so it is important to be able to identify those microorganisms and understand where they come from. Many pathogenic microorganisms can thrive in more than one environment. For instance, upon sampling the door handle of a refrigerator, one should expect to find enteric organisms such as gut microbiota, which live in the intestinal tracts of humans. Many of the natural flora that can be found in areas such as this are generally harmless to humans because they are a part of everyday life. However some of those organisms, such as certain serotypes of Escherichia coli can cause illness or even death when ingested by humans. It’s necessary to identify potential environments of harmful bacteria in order to prevent future cases of infection. In the case of E. coli types on high-traffic touching areas, proper hand washing is an aseptic technique utilized by many throughout the day and after using the restroom. If that refrigerator is located within a restaurant and it can be recognized that employees are not properly or irregularly washing their hands, an impending illness spawning from that restaurant can be easily avoided by a microbiologist regularly testing high-traffic areas used by employees such as the refrigerator.

In order to see what type of microbes live on or around the facial area, a sample was taken from the facial hair of a healthy male. The beards of healthy males surround mucus membranes such as the nose and mouth; these areas are breeding grounds, which provide warmth, moisture, and nutrients for microorganisms. A sample was taken from the beard of a male using a sterile swab and aseptic techniques to prevent contamination. That sample was grown and then isolated to a single microorganism for identification through analysis and biochemical testing.

 

Methods

A sample was obtained from the facial hair of an adult male. Using aseptic techniques, a sterile q-tip was dipped in sterile water and his beard was swabbed around the area of the mouth. After the specimen was obtained, the same q-tip was swabbed on a nutrient agar plate. The plate was placed in a warm area for three days and then incubated for two days. After noticeable growth was observed, a singular colony was obtained and streaked onto a new nutrient agar plate using aseptic technique and the quadrant streak plate method. The directions for quadrant streak plate method and all other procedures mentioned in this report can be found in the Leboffe Lab Manual (Leboffe, 2012). A Gram Stain was performed and the organism was viewed under a microscope to verify purity of the specimen. The pure specimen was then subjected to all required tests such as Catalase, Oxidase, Glucose Fermentation, Fluid Thioglycollate Broth, Motility, and Nitrate Reductase. In this case, additional staining and testing was needed such as an Acid Fast Stain, hemolysis on a blood agar plate, and Fructose Fermentation.

 

Results

The unknown was a Gram variable cocci with an opaque white pigmented colony. The unknown also had a convex elevation, smooth margin, and formed round colonies.

Major biochemical test results included Catalase, Fluid Thioglycollate Broth, acid fast staining, Nitrate Reductase, and Fructose Fermentation. The unknown tested negative for Catalase and was determined to be a facultative anaerobe. The unknown was also found to be negative for acid fast staining. Positive results were found for Nitrate Reductase and Fructose Fermentation (Table 1).

 

 

Table 1

Beard Specimen Biochemical Test Results

 

 

Test Performed

Test Result

Catalase

+

Oxidase

_

Glucose Fermentation

+

Fluid Thioglycollate Broth

Facultative Anaerobe

Motility

Non-motile

Nitrate Reductase

+

Gram Stain

Variable

Acid Fast Stain

_

Fructose Fermentation

+

Hemolysis

?

 

 

 

 

 

Discussion

The goal of the project was to obtain an environmental unknown, isolate it for purity, and administer biochemical testing in order to help identify it in Bergey’s Manual (Bergy, 1974). Initially the unknown was found to be a Gram positive cocci and a facultative anaerobe. This permitted the exclusion of all Gram negative and positive bacillus, and the inclusion of some Gram positive cocci with tendencies to also register as Gram variable. Since the unknown was also found to test negative for catalase, it needed to be Gram stained an additional time to confirm previous results, since a catalase negative organism would eliminate the possibility of the unknown to be apart of a larger group of Gram positive cocci. The catalase test results allowed for the exclusion of catalase positive Gram variable organisms such as all Gram variable rods, which test positive for Catalase. Also with the catalase test, it showed the unknown itself must be Gram variable, as Gram positive cocci which test negative for catalase were not present in Bergey’s manual. The second time the unknown was Gram stained, it was found to be Gram variable, not Gram positive. This was determined when the staining of the unknown was not consistent with the color produced after the Gram staining initially implemented. Gram variable cocci include species within the genus Gamella. The other species of Gamella were eliminated through their catalase results, which were positive. The catalase test was also administered a second time to verify that the organism does not produce the enzyme catalase. Organisms that do not produce the enzyme catalase must use terminal electron acceptors other than cytochrome C oxidase, which aerobic organisms use. The unknown organism was found to be positive for the nitrate reduction test. This verified that the unknown was able to transfer electrons to nitrate at the end of the Electron Transport Chain, demonstrating that the unknown respires anaerobically. The results of the nitrate and catalase tests were also agreed with by the Fluid Thioglycollate Broth, which showed that the unknown was a facultative anaerobe. The unknown organism grew both near the top of the test tube but also at the bottom where no oxygen was present. The FTB test showed that the organism had no oxygen requirement therefore was able to live in an anaerobic environment. An Acid Fast test was performed to eliminate all mycobacteria, which have cell walls composed of a waxy lipid. The results were negative, which confirmed the unknown did not have mycolic acid in its cell wall.

There are five species of the genus Gemella: Gemella haemolysans, Gemella morbillorum, Gemella bergeri, Gemella sanguinis, and Gemella palaticanis (Bergey, 1974). Only G. haemolysans and G. morbillorum are Gram variable and catalase negative so the other three were disregarded. Between G. haemolysans and G. morbillorum their differences between tests were for nitrate reductase and fructose fermentation. G. haemolysans was indeterminate for nitrate reductase in Bergey’s Manual, however for the same test G. morbillorum tested negative. As for fructose fermentation there was a more direct difference; G. haemolysans was shown to test positive for fructose fermentation, while G. morbillorum had tested negative. The unknown tested positive for fructose fermentation, therefore the unknown was identified to be most like G. haemolysans.

The scientific community has been aware of the Gemella species for nearly one hundred years. Gamella were first isolated from the throat, nose, and eyes in measles patients by Tunicliff in 1917 (Nalamada, 2010). It appears that G. haemolysans is commonly found around the facial area due to its proximity to the mouth and nose. It makes sense that the unknown was found to be most like the Gemella species, since this sample was taken from the facial area, close to the nose and mouth. Until recently, the Gemella species was sometimes difficult to differentiate. An unsolved case was for a twelve-year-old girl with a history of congenital heart disease (Purcell, 2001). Although doctors were able to confirm that the organism was most likely of the species Gemella, when the article, “Gemella species endocarditis in a child” by LK Purcell, JP Finley, R Chen, M Lovegren, and SA Halperin was written, they were unable to determine the specific type.

The name G. haemolysans sets it apart from others in that they are hemolysins. Hemolysins cause the lysis of red blood cells by destroying their cell membrane. The unknown, which was found to be most like G. haemolysins, are beta hemolytic, meaning “true hemolysins” (Table 1). In other more recent cases, G. haemolysans has been identified as the culprit of endocarditis. Endocarditis is an inflammation of the heart and is life threatening. In all of these cases however, the patients already had compromised immune systems before being diagnosed. Generally these patients are treated with antibiotic regimens to rid their bodies of the infection.

G. haemolysans is an opportunistic pathogen which can cause endocarditis and other severe infections (La Scola, 1998). It appears that G. haemolysans exists within the mucous membranes of healthy humans under normal circumstances because it has a low virulence. However if the individual carrying the bacteria is in poor health, gets sick or has surgery, G. haemolysans can cause one or more grave secondary infections. An example of this is a case of an elderly man with poor health practices who presented issues with eating and a high temperature. He was found to have an infection in his mouth, which lead to endocarditis caused by G. haemolysans infection. He was treated with antibiotics and his condition improved rapidly (La Scola, 1998).

Another infection caused by the Gemella species is endophthalmitis. Endophthalmitis is an inflammation of the eye. According to a scientific article called, “Acute postoperative endophthalmitis by Gemella haemolysansone,” by S Nalamada, S Jalali, and AK Reddy, of these complications was found after a man underwent surgery for cataracts. Cataracts are clouded growth areas in the lens of the eye or eyes of elderly patients or sometimes those with diabetes. They require surgical removal of the cataract and replacement of the lens. After surgery he got sick due to an infection of G. haemolysans. Like the other cases, he was treated with several antibiotics and topical steroid ointments (Nalamada, 2010).

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

Citations

Bergey, D. H., Buchanan, R. E., Gibbons, N. E., & American Society for Microbiology. (1974). Bergey's manual of determinative bacteriology. Baltimore: Williams & Wilkins

 

La Scola, B., & Raoult, D. (1998). Molecular Identification of Gemella Species from Three Patients with Endocarditis. Journal of Clinical Microbiology, 36(4), 866–871.

 

Leboffe, M. J., & Pierce, B. E. (2012). Microbiology: Laboratory theory & application. Englewood, CO: Morton Pub.

 

Nalamada, S., Jalali, S., & Reddy, A. K. (2010). Acute postoperative endophthalmitis by Gemella haemolysans. Indian Journal of Ophthalmology, 58(3), 252–253. http://doi.org/10.4103/0301-4738.62658

 

Purcell, L. K., Finley, J. P., Chen, R., Lovgren, M., & Halperin, S. A. (2001). Gemella species endocarditis in a child. The Canadian Journal of Infectious Diseases, 12(5), 317–320.


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