Klebsiella pneumoniae as an Etiological Factor of Bovine Mastitis in Puerto Rico
Board Location: #15
Discipline: Biological Sciences
Subcategory: Microbiology/Immunology/Virology
Session: 3
Debora Torres Centeno - University of Puerto Rico - Aguadilla Campus
Co-Author(s): Stephanie González Avilés, University of Puerto Rico in Aguadilla, PR; Wilgeyshka Acevedo Acevedo University of Puerto Rico in Aguadilla, PR; Linda Hernández Pérez University of Puerto Rico in Aguadilla, PR; Javier Galarza Avilés University of Puerto Rico in Aguadilla, PR; Yavier Nazario-Cáceres, University of Puerto Rico in Aguadilla, PR; Zaidlys Hernández Vargas, University of Puerto Rico in Aguadilla, PR; Xiger E. González García, University of Puerto Rico in Aguadilla, PR; Gloribel Seguí Méndez, University of Puerto Rico in Aguadilla, PR; Guillermo Ortiz-Colón, University of Puerto Rico in Mayagüez, PR; and Juliana Pérez Laspiur, University of Puerto Rico in Aguadilla, PR
Klebsiella pneumoniae is a gram-negative, lactose-fermenting, facultative anaerobic, rod-shaped bacteria known for being a common etiological factor of clinical and subclinical mastitis in the mainland. A previous pilot study conducted at a college farm in USA, demonstrated that K. pneumoniae was one of the most prevalent pathogens causing mastitis at that location. Correct and fast identification of the bacteria causing mastitis is essential for successful animal treatment. Mastitis is responsible for approximately 20 million dollars loss annually in the dairy industry because it presents impaired milk production and quality. The objective of this study was to develop a test for the identification of Klebsiella Pneumoniae in milk samples from cows suspected of mastitis in the south-western region of Puerto Rico, to determine its prevalence and provide targeted treatment for this condition by using microbiology and molecular biology techniques. For this, milk samples were collected from 54 cows with suspected mastitis and diagnosed using the CMT and PortaSCC assays. Samples were inoculated in MacConkey agar at 37° for 24-48 hours, followed by biochemical testing, including oxidase, catalase, urease, pyrase and coagulase assays. Genetic identification was performed by using PCR analysis targeting the 16s rRNA gene. Results are immediately communicated to the farmer to customize animal treatment. Preliminary data suggests that 2 of 19 cows with mastitis are infected with K. pneumoniae, which indicates a low prevalence (11%) of the bacteria in the herd. Future research involves testing additional samples, evaluating the hemolytic activity of K. pneumoniae by using blood agar, developing quantitative real-time PCR for immediate identification of K. pneumoniae, and testing for antibiotic resistance.
Funder Acknowledgement(s): MSEIP SOLARIS. Minority Science and Engineering Improvement Program (MSEIP)-Supporting Opportunities for Learning, Awareness, Research, and Inclusion in STEM and Citizen Science (SOLARIS). United States Department of Education Title III Grant No. P120A230084 awarded to the University of Puerto Rico-Aguadilla. / Academic Research and Enhancement Award (AREA) Program. NIH Eunice Kennedy Shriver National Institute of Child Health and Human Development R15 AREA Program Grant No. 1R15HD109919-01
Faculty Advisor: Juliana Pérez-Laspiur, PhD, juliana.perez-laspiur@upr.edu
Role: Milk collection, California Mastitis Test (CMT) and Porta SCC tests performance, agar preparation, milk inoculation, subcultures, gram staining, biochemical testing, primer designing for target gene, bacterial scrapping with PBS, reagent preparation, genomic DNA extraction, PCR, electrophoresis of nucleic acids in agarose gel, data collection, statistical analysis, individual and group work, among other responsibilities.