Abstract #305
Section: Physiology and Endocrinology
Session: Physiology & Endocrinology III
Format: Oral
Day/Time: Tuesday 10:00 AM–10:15 AM
Location: 330
Session: Physiology & Endocrinology III
Format: Oral
Day/Time: Tuesday 10:00 AM–10:15 AM
Location: 330
# 305
Discovering neutrophil extracellular traps in the bovine endometrium and the effects of feeding a rumen-protected methionine on plasma amino acid concentrations and uterine characteristics.
S. L. Stella*1, D. A. V. Acosta2, C. Skenandore1,3, Z. Zheng1, A. Steelman1, D. Luchini4, F. C. Cardoso1, 1University of Illinois, Urbana, IL, 2The Colombian Corporation for Agricultural Research (CORPOICA), Bogotá, Colombia, 3Texas A&M College of Veterinary Medicine, College Station, TX, 4Adisseo NACA, Alpharetta, GA.
Key Words: rumen-protected methionine, PMN, neutrophil extracellular traps
Discovering neutrophil extracellular traps in the bovine endometrium and the effects of feeding a rumen-protected methionine on plasma amino acid concentrations and uterine characteristics.
S. L. Stella*1, D. A. V. Acosta2, C. Skenandore1,3, Z. Zheng1, A. Steelman1, D. Luchini4, F. C. Cardoso1, 1University of Illinois, Urbana, IL, 2The Colombian Corporation for Agricultural Research (CORPOICA), Bogotá, Colombia, 3Texas A&M College of Veterinary Medicine, College Station, TX, 4Adisseo NACA, Alpharetta, GA.
Supplementing methionine, the most limiting amino acid (AA) to dairy cows (NRC 2001), may improve uterine health and reveal that other AA concentrations have been affected. The objective of this study was to observe the effects of feeding rumen-protected methionine (RPM; Smartamine M) on plasma AA concentrations, uterine cytology, neutrophil counts, and to confirm neutrophil extracellular trap (NET) formation in the bovine endometrium. Multiparous Holstein cows (n = 20) were randomly assigned to 2 treatments starting 21d before calving until 73DIM. Treatments were: CON (n = 9, TMR with a Lys:Met = 3.5:1) and MET (n = 11, TMR+RPM with a Lys:Met = 2.8:1). Uterine endometrial biopsies and blood samples from the coccygeal artery/vein were obtained at 15, 30, and 73DIM. Biopsy samples were sectioned and stained using immunohistochemistry with Hoechst (DNA) and Anti-Neutrophil Elastase antibody (NE). Biopsy slides were scanned in an automated imaging cytometer to quantify neutrophil numbers and a confocal fluorescent microscope for NET discovery/confirmation via NE and DNA fluorescent antibodies. Endometrial swabs were streaked onto slides, stained with Giemsa, and scanned using whole image scanning. Polymorphonuclear neutrophils (PMN) were counted and a percentage was calculated based on the number of PMN to epithelial cells. Statistical analysis was performed using the MIXED procedure of SAS. CON had lower (P < 0.01) methionine plasma concentrations (18.05 ± 2.0μM/mL) than MET (30.39 ± 1.6μM/mL). CON had higher (P < 0.01) cystine plasma concentrations (3.62 ± 0.3μM/mL) than MET (2.8 ± 0.3μM/mL). A treatment by DIM interaction was observed for PMN and the number of neutrophils in the endometrium: CON (28.28 ± 7.7%) tended to have higher (P = 0.09) PMN percentage in swabs than MET (18.19 ± 6.7%) and CON (1423.98 ± 437.9) tended to have higher (P = 0.06) neutrophil numbers in the endometrium than MET (1192.54 ± 408.2). Supplementation of RPM appears to alter the concentrations of AA and have beneficial effects on uterine immune function.
Key Words: rumen-protected methionine, PMN, neutrophil extracellular traps
