122 Response of Ruminant Neutrophils on Fresh Filtered Garlic Preparation

Abstract

Abstract The purpose of this study was to evaluate the effect of fresh filtered garlic preparation on ruminant neutrophils in-vitro. Blood was collected from the jugular vein of clinically healthy female St Croix sheep (n = 3) and Boer x Spanish goats (n = 3) at NC A&T State University Farm. Neutrophils were isolated from 10 mL whole blood sample of each animal. Fresh filtered garlic were prepared to stimulate the neutrophils. Before treating the neutrophils, the total cell count was measured, and 1 million cells were taken for treatment. Total protein concentration at different dilution levels of fresh filtered garlic were analyzed using BCA assay for the volumetric determination of garlic to be used at 10 ug/mL concentration. Before and after treatment the total protein concentration in the neutrophil supernatant were also measured. RNA was extracted from treated and untreated neutrophils pellet using TRIZOL RNA Isolation Protocol. The pooled RNA of goat and sheep samples were used for cDNA synthesis followed by qPCR analysis of galectin (Galectin-1, -3, -9) gene modulation in relation to indicators of animal health and production. Due to treatment, the total protein concentrations were increased. The magnitude of galectin gene expression was also observed in both sheep and goat neutrophils treated with fresh filtered garlic preparation. Galectin-3 and -9 expression increased manifolds in both sheep and goat neutrophils as compared with the control (untreated neutrophils) whereas galectin-1 increased in goat but decreased in sheep neutrophils. These results indicate that garlic had no adverse effect and differentially modulated galectin secretion in vitro. Galectins regulate biologic functions, immunity and disease pathogenesis and can serve as biomarkers. Further studies are warranted to determine the health and production benefits of the observed galectin modulation using garlic in small & large ruminants.