Abstract
Antibiotic resistance represents a significant global health crisis, especially within hospitals, creating a necessity for the search for alternative therapeutic agents to combat drug-resistant pathogens. One alternative treatment is phages specifically targeting G. rubripertincta, a gram-positive, aerobic actinomycete bacterium which has shown intrinsic resistance to some antibiotics. Furthermore, G. rubripertincta can cause endocarditis and CNS infections in immunocompromised individuals. Several phages such as Alyssamiracle, Fribs8, Genamy16, and NovaSharks, have been discovered that successfully target G. rubripertincta. The goal of this research is to determine the optimal lysis time and multiplicity of infection (MOI) of these phages. The phages work in a lytic manner, bursting the bacteria and releasing its viral progeny, further enhancing the targeting of the bacteria. The lysis time and optimal MOI were determined by growing G. rubripertincta with different amounts of phage. The absorbance readings were taken from triplicate trials using a 96-well microplate reader at OD600 every 10min for a total of 48h. Results showed that the higher the MOI, the quicker the lysis time. For both Alyssamiracle and Fribs8, an MOI of ~100 had the quickest lysis time of 4.4h. The growth rate after lysis was measured and showed that the higher the MOI, the higher the growth rate, suggesting a faster recovery possibly due to lysogens or resistant bacteria emerging. As aforementioned, with G. rubripertincta’s inherent resistance to antibiotics, knowing the phages ideal MOI and lysis time will aid in the development of more effective phage therapy.
Faculty Sponsors
Dr. Robert Smith, Dr. Julie Torruellas Garcia
Project Type
Event
Location
Alvin Sherman Library
Start Date
4-2-2025 12:45 PM
End Date
4-3-2025 12:00 AM
Determining the Optimal Multiplicity of Infection (MOI) and Lysis Time of Gordonia rubripertincta Bacteriophage
Alvin Sherman Library
Antibiotic resistance represents a significant global health crisis, especially within hospitals, creating a necessity for the search for alternative therapeutic agents to combat drug-resistant pathogens. One alternative treatment is phages specifically targeting G. rubripertincta, a gram-positive, aerobic actinomycete bacterium which has shown intrinsic resistance to some antibiotics. Furthermore, G. rubripertincta can cause endocarditis and CNS infections in immunocompromised individuals. Several phages such as Alyssamiracle, Fribs8, Genamy16, and NovaSharks, have been discovered that successfully target G. rubripertincta. The goal of this research is to determine the optimal lysis time and multiplicity of infection (MOI) of these phages. The phages work in a lytic manner, bursting the bacteria and releasing its viral progeny, further enhancing the targeting of the bacteria. The lysis time and optimal MOI were determined by growing G. rubripertincta with different amounts of phage. The absorbance readings were taken from triplicate trials using a 96-well microplate reader at OD600 every 10min for a total of 48h. Results showed that the higher the MOI, the quicker the lysis time. For both Alyssamiracle and Fribs8, an MOI of ~100 had the quickest lysis time of 4.4h. The growth rate after lysis was measured and showed that the higher the MOI, the higher the growth rate, suggesting a faster recovery possibly due to lysogens or resistant bacteria emerging. As aforementioned, with G. rubripertincta’s inherent resistance to antibiotics, knowing the phages ideal MOI and lysis time will aid in the development of more effective phage therapy.
