Tuesday, March 3, 2015

Positively Gram-positive: Gram-staining Results, Comparisons, & Observations

After Gram-staining our mystery bacteria last week, we initially suspect that our bacteria is Gram-positive due to many observed characteristics. Below is a photograph through a microscope of what we are basing our observations on. Our bacterium appears to be stained purple, which is a result of uptake of crystal-violet stain. The purple stain indicates a thick layer of peptidoglycan in the cell wall. The bacterium possesses a diplobacilus rod shape. 

Gram staining results


To verify our results, we used a MacConkey agar to determine whether our mystery bacterium found in the soil is Gram-negative or Gram-positive. We compared our mystery bacterium to three controls: Bacillus megaterium, which was a Gram-positive control, Pseudomonas aureoygynosa, which was a Gram-negative, non-lactose fermenting control, and Klebsiella pneumonia, a Gram-negative, lactose-fermenting control. Below is a photograph of our MacConkey agar, showing our results for growth on the agar for our controls and our bacterium. Our bacterium, according to our results on the MacConkey agar, is Gram-positive, indicating that our bacterium did not grow on the agar. 


MacConkey agar 

Gram-positive cell walls have a thick layer of peptidoglycan with teichoic acids, a polyalcohol. On the other hand, the cell wall of a bacterium that is Gram-negative will have a thin layer of peptidoglycan, with a bilayer membrane on the outside of the peptidoglycan. This bilayer membrane is composed of phospholipids, proteins, and lipopolysaccharides.

Although we are concerning ourselves here with bacterium found in soil outside, Gram-negative bacteria and Gram-positive bacteria are prevalent in our every day society. These bacterium are the cause of common infections found in almost all hospitals. Gram-negative bacterium cause many infections, such as pneumonia, bloodstream infections, wound infections, surgical site infections, and meningitis. Gram-negative bacteria exhibit multi-drug resistance to many antibiotics (CDC, 2011; Moehring & Anderson, 2014). According to the CDC, Gram-negative bacteria have built-in systems that are able to be resistant and are able to pass along genetic information that will allow other bacteria to also become drug-resistant. Antibiotics for Gram-negative bacterial infections differ from those of Gram-positive due to the risk of septic shock, morbidity, and mortality in patients. Gram-positive bacteria also pose a threat as they are becoming increasingly resistant to antibiotics used to treat infections caused by Staphylococcus aureus and Enterococcus spp. (Woodford & Livermore, 2009). More commonly known as MRSA, this Gram-positive bacteria spreads rapidly throughout hospitals. Both Gram-positive and Gram-negative exhibit multidrug resistance to antibiotics and antimicrobials aimed to control and eliminate bacterial infections.

“Gram-negative Bacteria Infections in Healthcare Settings.” Centers for Disease Control and Prevention. Centers for Disease Control and Prevention, 17 Jan. 2011. Web. 02 Mar. 2015.


Moehring, R., D. Anderson, “Gram-negative Bacillary Bacteremia in Adults” Gram-negative Bacillary Bacteremia in Adults. N.p., 31 Oct 2014. Web. 02 Mar. 2015.


Woodford, N., DM. Livermore, (2009) Infections caused by Gram-positive bacteria: a review of the global challenge. J Infect. 59, 60003-7.


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