Low-density dispersive liquid-liquid microextraction with a subsequent esterification step was proposed in this study for the determination of linoleic acid and stearic acid in sheep blood serum samples. The method developed aimed to quickly and efficiently extract and esterify both fatty acids with subsequent analysis by gas chromatography/flame ionization detection. The extraction method was optimized with Scheffé’s polynomial model for three-component mixtures. The method was validated according to the parameters established by Eurachem Guide (2014). The optimal extraction conditions for LD-DLLME were 1400 µL of dispersion medium (MgCl2 0.017%), 400 µL of extractor solvent (toluene) and 1200 µL of dispersion solvent (methanol). The method performance showed adequate selectivity, sensitivity and precision to be applied to real samples, with an average recovery of 98.54% for linoleic acid and 103.83% for stearic acid. LD-DLLME was superior to the traditional method of analysis, which has been used until now for the determination of fatty acids in blood serum samples from ruminants. The analysis of real samples showed that the developed method is efficient for monitoring these substances in ruminants.

ABSTRACT The objective of this study was to evaluate the effect of replacing ground corn (402 g kg−1 of dry matter) with rehydrated corn grain silage (RCGS; 425 g kg−1 of dry matter) in a flushing diet on follicular development in tropical Santa Inês ewes. Fifteen ewes were randomly assigned to one of two treatment groups: a diet with ground corn (control, n = 7) or a diet with RCGS (n = 8). The first day of the diets was designated d0, and the diets were fed for 30 days, up to two days after the end of the estrus synchronization protocol. The estrus synchronization protocol (intravaginal progestogen sponge for 11 days plus 300 IU eCG and 37.5 µg d-cloprostenol IM two days before sponge removal) started on d17 of the diets. The number and diameters of ovarian follicles ≥3 mm were assessed by ultrasound on the day before the diets were provided (d−1), on d14, and then daily from two days before sponge removal until ovulation or up to the eighth day after sponge removal. Blood samples were collected on days −1, 0, 6, 12, 18, 24, and 30 for glucose and urea analyses. There was no effect of the treatments on dry matter intake, plasma glucose, and urea nitrogen concentrations, or on the percentage of ewes that ovulated and on the number of ovulations. The number of follicles ≥3 mm did not differ between treatment groups; the number increased between d−1 and d14 and did not differ between d14 and d28. The diameter of ovulatory follicles at sponge removal, the interval from sponge removal to estrus, and the growth rate of the ovulatory follicles were greater in the RCGS group than in the control group, resulting in larger follicles at ovulation. Replacement of ground corn by RCGS in the flushing diet does not increase the number of ovulations but results in delayed onset of estrus and ovulation of larger follicles.