MK-212 increases anxiety and decreases locomotor activity. The anxiogenic-like profile of 5-HT2 receptor activation is prevented by the blockade of 5-HT2 receptors within the BLA, which does not have an effect by itself upon basal anxiety levels triggered by the EPM.
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.
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The goal of this study was to evaluate the effects of different levels (10 and 13% crude protein) and protein source on ruminal fermentation of feedlot Nellore steers fed high-concentrate diets. The protein sources tested were urea, dried distillers grains (DDG) and corn gluten meal. The DDG and gluten were used as RUP sources. We used 6 cannulated Nellore steers, rumen, duodenum, and ileum arranged in 6×6 Latin square designed in factorial (2 nitrogen levels and 3 protein sources) balanced for residues. The pH was measured immediately after rumen fluid sampling at 0, 2, 4, 6, 8 and 10h after feeding. The NH3-N concentrations were determined with micro-Kjeldahl apparatus. The concentration of volatile fatty acids (VFA) was assessed by gas chromatography. Data were evaluated using the MIXED procedure of SAS, with repeated measures over time. There was no interaction between sampling time and treatments for NH3-N and pH (P > 0.05). Animals fed diets containing Urea had a higher concentration of NH3-N (20.6 mg/dL) than animals supplemented with Gluten (16.2 mg/dL) or DDG (12.7 mg / dL). There was an increase in the NH3-N concentration (P = 0.049) with the increase of the protein level 10 to 13% CP with 14.7 and 18.3 mg/dL, respectively. The ruminal pH was affected by the dietary protein level (P = 0.016), in which animals fed a level of 10% CP had a higher pH (6.80) compared to the level of 13% CP (6.43). Total and individual VFA concentration was not affected by protein levels and sources (P > 0.05). The reduction of N level and the use of RUP did not adversely affect the ruminal fermentation. Although significant differences were observed for pH, they were small and adequate for microbial protein synthesis in the rumen.
The modification in the nutritional composition of the ruminant diet causes significant alterations in the fatty acids (FAs) structure supplemented because of the action of rumen microorganisms. The modification in the FAs structure alters the role that these play in the ruminant metabolism. The Folch method is the most often used to determine fatty acids in these animals' tissues and presents certain disadvantages such as the great volume of solvent and low mass transfer from the analyte to the extracting phase. Thus, we tested the low-density dispersive liquid-liquid microextraction (LD-DLLME) as an alternative method to determine these substances. In this paper, a simplesaugmented mixture design was used. The Scheffé's polynomial was applied in that design, and Moore-Penrose generalized matrix inverse was used because of the possibility of concurrently determining value estimates of coefficients of the parameters that represent cubic terms. The application of the modeling allowed the chemical interpretation of the LD-DLLME best extraction condition for linoleic acid in ruminant serum samples.
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