Discussion After doing a series of different biochemical tests for the isolated organisim from the sputum sample,

it was concluded that the unknown Gram positive organism was Streptococcus pyogenes. The sputum sample was first inoculated in the agar plates MAC, SBAP, CHOC and also a direct gram stain of the sample was performed. After a day of incubation, the MAC agar gave a negative result, therefore it is not a Gram-negative bacteria and non-lactose fermenting. The SBAP and CHOC showed growth of colonies. The CHOC agar had a mixed formation of colonies (gray colonies and 2+ tan colonies), so it was isolated in another CHOC agar and it was mixed again. A gram stain procedure was done and it showed both Gram positive and negative cocci. The tan colonies from the CHOC agar were inoculated to SBAP 1. From the direct culture of the sputum sample to SBAP there was formation of 2+ -hemolytic colonies and 2+ B hemolytic colonies. Each was isolated on different plates to have a primary culture, SBAP 1 and SBAP 2. In SBAP 1, a split plate was done, by inoculating the B hemolytic colonies and the tan colonies from the CHOC. It gave a mixed growth, so the tan colonies were inoculated to SBAP 2 in order to isolate the tan colonies without being mixed with the other colony. In SBAP 2, the split plate of hemolytic colonies and tan colonies from the CHOC. After incubation, the tan colonies showed irregular colony morphology which has a formation of halo clearing on the sides of the colony. So the tan colony with irregular colony morphology was further isolated in SBAP 3 which showed a pure B hemolytic, Gram positive cocci in chains after incubation. The colony was worked-up; the catalase test gave a negative result, meaning the bacteria did not produced the enzyme catalase capable of destroying the hydrogen peroxide[3]. confirming that the unknown bacterium it is a Streptococcus spp. Bacitracin susceptibility test was done and it showed a 24 cm diameter of clearing, there a susceptible result. The group decided to work-up until bacitracin susceptibility test to confirm if the organism is Streptococcus pyogenes or Streptococcus agalactiae. Bacitracin is an antibiotic interfering with the synthesis of peptidoglycan, a major component of bacterial cell walls. Different types of bacteria have different degrees of susceptibility to bacitracin. This test determines whether the bacterium is either sensitive (susceptible) to bacitracin or resistant to the drug.[2] Beta-hemolytic streptococci produce a toxin that forms a clear zone of hemolysis on blood agar, demonstrating its ability to destroy red blood cells. This hemolysis is attributed to toxins formed by Group A streptococci called streptolysins. Streptolysins can destroy not only red blood cells, but also the white blood cells responsible for fighting off bacteria and disease, as well as other body cells[2]. The cause of strep throat is bacteria known as Streptococcus pyogenes, also known as group A streptococcus. Streptococcal bacteria are highly contagious. They can spread through airborne droplets when someone with the infection coughs or sneezes, or through shared food or drinks. You can also pick up the bacteria from a doorknob or other surface and transfer them to your nose, mouth or eyes.[3]

References: [1]Bacitracin Susceptibility Test. (n.d.).Intuitive Systems, Inc. - Makers of Virtual Laboratory Software. Retrieved October 1, 2013, from http://www.vumicro.com/vumie/help/VUMI [2]Catalase Test. (n.d.). Intuitive Systems, Inc. - Makers of Virtual Laboratory Software. Retrieved October 2, 2013, from http://www.vumicro.com/vumie/help/VUM [3]Strep throat: Causes - MayoClinic.com. (n.d.). Mayo Clinic. Retrieved October 1, 2013, fromhttp://www.mayoclinic.com/health/strepthroat/DS00260/DSECTION=causes [4]Tortora G, Funke B, Case C. 2007. Microbiology: An Introduction, 9th Edition. Pearson Education, Inc.