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ARS Home » Northeast Area » Beltsville, Maryland (BARC) » Beltsville Agricultural Research Center » Animal Genomics and Improvement Laboratory » Research » Publications at this Location » Publication #316236

Research Project: Understanding Genetic and Physiological Factors Affecting Nutrient Use Efficiency of Dairy Cattle

Location: Animal Genomics and Improvement Laboratory

Title: The effect of lipopolysaccharide (LPS) and phorbol 12-myristate 13-acetate (PMA) on whole blood oxidative response as assessed by luminol-amplified chemiluminescence in dairy cows

Author
item Qu, Yang - University Of Maryland
item Kahl, Stanislaw - Stass
item Connor, Erin
item Elsasser, Theodore
item Moyes, Kasey - University Of Maryland

Submitted to: Journal of Animal Science
Publication Type: Abstract Only
Publication Acceptance Date: 4/20/2015
Publication Date: 7/12/2015
Citation: Qu, Y., Kahl, S., Connor, E.E., Elsasser, T.H., Moyes, K.M. 2015. The effect of lipopolysaccharide (LPS) and phorbol 12-myristate 13-acetate (PMA) on whole blood oxidative response as assessed by luminol-amplified chemiluminescence in dairy cows. Journal of Animal Science. 126.

Interpretive Summary:

Technical Abstract: The differences between lipopolysaccharide (LPS) and phorbol 12-myristate 13-acetate (PMA) on whole blood oxidative response using luminol-amplified chemiluminescence (CL) are currently unknown in cattle. Luminol-dependent CL measures the amount of reactive oxygen species released from leukocytes after stimulation with LPS and/or PMA. The objective of this study was to compare in vitro the effects of PMA and LPS on oxidative response in whole blood of dairy cows during lactation as a rapid means of assaying the oxidative response of blood leukocytes. Jugular blood (20 mL) was collected from 6 healthy multiparous Holstein dairy cows in mid-lactation (> 90 DIM) using EDTA vacutainer tubes. For each cow, 500 microliters of blood was incubated at final concentrations of either 0, 200, 800 or 1600 ng/mL of PMA or LPS for 15 min at 37 degrees Celcius using a heating block. After incubation, oxidative response of whole blood was measured using a chemiluminometer. Data were analyzed by ANOVA using the PROC MIXED procedure of SAS. Overall, whole blood incubated with PMA resulted in higher (P < 0.001) CL values (800 ng/mL; 2635 relative units) than LPS (800 ng/mL; 777 relative units). In PMA, a significant dose response relationship was observed where incubation with 200, 800 or 1600 ng/mL resulted in progressively higher CL values than 0 ng/mL. In addition, incubation with PMA resulted in higher CL values when compared with LPS. In conclusion, although both LPS and PMA generated an oxidative response measurable by CL, PMA elicited a CL response greater than that of LPS. The data suggest that PMA stimulation of cells in whole blood may serve as a rapid test of oxidative burst responsiveness to assess a vital aspect of immune function in dairy cows.