Isolation and characterization of Mycoplasma bovis in milk samples from cows with clinical mastitis.
Board Location: #148
Discipline: Biological Sciences
Subcategory: Microbiology/Immunology/Virology
Session: 2
Yavier Nazario - University of Puerto Rico, Aguadilla
Co-Author(s): Wilgeyshka S. Acevedo Acevedo, University of Puerto Rico in Aguadilla, Aguada, Puerto Rico
Clinical mastitis is the number one cause of cessation of milk production in both, humans and cows. Mastitis can be caused by several bacteria that stimulate different immune responses by the lactating mammal and require customized treatment with specific antibiotics. Identifying which bacteria is causing clinical mastitis in an animal is essential to prescribe an effective treatment for fast recovery and no loss of milk production. Mycoplasma bovis is a gram-negative bacteria that is known to cause clinical mastitis and other serious diseases in cows on the mainland. However, in Puerto Rico, we consider other bacteria such as as E. coli and S. aureus to be primarily responsible for this disease. Investigating the prevalence of M. bovis in cows with mastitis is important because it is necessary for implementing the best preventive and treatment management practices to prevent the further spreading of this bacterium to other animals on the farm. The objective of this study is to investigate the prevalence of M. bovis in milk samples from dairy cows with clinical mastitis in farms in the southwest of Puerto Rico using microbiology and molecular biology techniques. Milk samples will be collected from cows diagnosed with clinical mastitis, transported to the laboratory, enriched in selective media for gram-negative bacteria, and incubated at 37°C for 48 hours. Colony count and morphological analysis are then performed to select suspected M. bovis colonies based on known characteristics. Biochemical tests and PCR amplification of target gene polC will be conducted to identify M. bovis in milk samples. The prevalence of M. bovis in milk samples from cows with clinical mastitis will be calculated and reported to each farm at the end of the trial. Preliminary results are yet to be found. The control group used is milk samples from healthy cows. References:Gelgie AE, Korsa MG, Kerro Dego O. Mycoplasma bovis Mastitis. Curr Res Microb Sci. 2022 Feb 24;3:100123. doi: 10.1016/j.crmicr.2022.100123. PMID: 35909617; PMCID: PMC9325741.Hashem YM, Mousa WS, Abdeen EE, Abdelkhalek HM, Nooruzzaman M, El-Askary A, Ismail KA, Megahed AM, Abdeen A, Soliman EA, Wareth G. Prevalence and Molecular Characterization of Mycoplasma Species, Pasteurella multocida, and Staphylococcus aureus Isolated from Calves with Respiratory Manifestations. Animals (Basel). 2022 Jan 27;12(3):312. doi: 10.3390/ani12030312. PMID: 35158636; PMCID: PMC8833736.
Funder Acknowledgement(s): This research project was supported by PR-LSAMP Undergraduate Research Scholarships, RUTAS Program USDA Grant Award #2018-38422-28570 and AREA R15 NIH Grant Award #1R15HD109919-01 granted to Juliana Perez Laspiur Ph.D. University of Puerto Rico at Aguadilla, Aguadilla, PR.
Faculty Advisor: Juliana Pérez Laspiur, juliana.perez-laspiur@upr.edu
Role: Milk collection, non-selective and selective agar preparations, biochemical tests, genomic DNA isolation, primer design for target gene, DNA bioinformatics, PCR, Real-time PCR, and electrophoresis of nucleic acids. I also learned to prepare reagents and materials for this study, follow research protocols, work individually and in groups, learn to collect data, basic statistical analysis, and prepare written and oral research reports, among other skills.