Medium-chain Fatty Acids Research Articles

#2765 Medium chain fatty acids are potent binding competitors to improve protein-bound uremic toxin clearance in hemodialysis

Abstract Background and Aims Protein-bound uremic toxins (PBUTs) remain a concerning burden in patients with end-stage renal disease (ESRD) since their removal during hemodialysis is limited due to their tight binding to albumin. By increasing their circulating free fraction during HD, “binding competitors” of albumin could be used to increase their dialytic clearance. Medium chain fatty acids (MCFAs, 6 to 12 carbon length) are potent candidates for that purpose. The present study aims to investigate whether MCFAs could serve as potential binding competitors of albumin to enhance the removal of PBUTs, specifically IS and p-CS, during hemodialysis. Method Hexanoate (C6), octanoate (C8), and decanoate (C10) were purchased from Sigma-Aldrich. The binding of MCFAs to albumin was investigated in silico and using fluorescent probes: warfarin (specific of Sudlow's site I) and dansylsarcosine (specific of Sudlow's site II). Indoxyl-sulfate (IS, final concentration 212 µM) and p-cresyl sulfate (p-CS, final concentration 250 µM) were added to a 600 µM bovine serum albumin-phosphate buffered saline solution. The free fractions of IS and p-CS were assayed with or without MCFAs (1-2 mM) using ultrafiltration devices in this solutions and in patients uremic plasma. To mimic the removal of PBUTs during a HD session, batches of 2 liters of fresh bovine blood were loaded with IS and p-CS (final concentration 200 µM). A 2-hours closed-loop HD session was performed using a Fresenius 5008 CorDiax HD generator (Fresenius, Germany). A solution of 224 mM of octanoate was perfused at the rate of 150 µL/min; A solution of saline 0.9% was used as a control. One milliliter of blood was sampled every 15 minutes through the arterial sampling port and concentration of IS and p-CS were assayed by HPLC coupled with fluorescence detection. The hemolytic effect of MCFAs was evaluated in vitro by assaying the free hemoglobin concentration. Results According to the in silico prediction, the binding was predicted to be almost complete for MCFAs ≥ C8 (Fig. 1). The enhanced protein binding appeared to be inversely correlated with water solubility, which became particularly low for MCFAs ≥ C10. Octanoate and decanoate were the more prone to displace dansylsarcosine from Sudlow's site II of albumin (which is the main binding site of IS and p-CS). In vitro, the incubation with 2 mM of octanoate and decanoate increased the free fraction of PBUTs from 12% to 53% for p-CS (4.4 fold, p < 0.05) and from 11% to 45% for IS (4.1 fold, p < 0.05). Octanoate was also able to increase the free fraction of PBUTs in patients uremic plasma (1.6-fold for IS, p < 0.05 and 4.8 fold for p-CS, <0.01). The per-dialytic infusion of octanoate significantly increased the fractional removal of p-CS (from 38% to 88%, p < 0.001) and IS (from 36% to 91%, p < 0.001). No significant hemolysis was observed for concentration of MCFAs lower than 2 mmol/L. Conclusion MCFAs and especially octanoate and decanoate are serious candidates to displace the binding of PBUTs such as p-CS and IS. The per-dialytic administration of octanoate significantly increased the removal of PBUTs and could constitute a new strategy to prevent their accumulation in end-stage kidney disease patients. Due to their good safety profile, MCFAs could be better tolerated than other chemical compounds that has already been tested in a clinical setting such as ibuprofen. Further in vivo studies are however needed to carefully evaluate this potentially new therapeutic option.

Whole-cell one-pot biosynthesis of dodecanedioic acid from renewable linoleic acid

BackgroundDodecanedioic acid (DDA), a typical medium-chain dicarboxylic fatty acid with widespread applications, has a great synthetic value and a huge industrial market demand. Currently, a sustainable, eco-friendly and efficient process is desired for dodecanedioic acid production.ResultsHerein, a multi-enzymatic cascade was designed and constructed for the production of DDA from linoleic acid based on the lipoxygenase pathway in plants. The cascade is composed of lipoxygenase, hydroperoxide lyase, aldehyde dehydrogenase, and unidentified double-bond reductase in E. coli for the main cascade reactions, as well as NADH oxidase for cofactor recycling. The four component enzymes involved in the cascade were co-expressed in E. coli, together with the endogenous double-bond reductase of E. coli. After optimizing the reaction conditions of the rate-limiting step, 43.8 g L− 1 d− 1 of DDA was obtained by a whole-cell one-pot process starting from renewable linoleic acid.ConclusionsThrough engineering of the reaction system and co-expressing the component enzymes, a sustainable and eco-friendly DDA biosynthesis route was set up in E. coli, which afforded the highest space time yield for DDA production among the current artificial multi-enzymatic routes derived from the LOX-pathway, and the productivity achieved here ranks the second highest among the current research progress in DDA biosynthesis.Graphical

Deciphering the reduction of antibiotic resistance genes (ARGs) during medium-chain fatty acids production from waste activated sludge: Driven by inhibition of ARGs transmission and shift of microbial community

Medium-chain fatty acids (MCFAs) production from waste activated sludge (WAS) by chain extension (CE) is a promising technology. However, the effects and mechanisms of CE process on the fate of antibiotic resistance genes (ARGs) remain unclear. In this study, the results showed that the removal efficiency of ARGs was 81.15 % in CE process, suggesting its efficacy in reducing environmental risks. Further, the observed decrease in mobile genetic elements (MGEs) indicated that CE process restricted the horizontal gene transfer (HGT). Complementing this, the increase in soluble organic matters and extracellular 16 S rDNA confirmed that MCFAs production caused bacterial damage. Decreased intracellular ARGs and increased extracellular ARGs further revealed that MCFAs production impaired ARGs hosts, thereby limiting the vertical gene transfer (VGT) of ARGs. Shift of microbial community combined with co-occurrence network analysis demonstrated that functional bacteria without host potential for ARGs were enriched, but potential ARGs and MGEs hosts decreased, showing the role of functional bacterial phylogeny and selection pressure of MCFAs in reducing ARGs. Finally, partial least squares path model was used to systematic verify the mechanism of ARGs removal in CE process, which was attributed to the inhibition of ARGs transmission (HGT and VGT) and shift of microbial community.

Case for supporting astrocyte energetics in glucose transporter 1 deficiency syndrome.

In glucose transporter 1 deficiency syndrome (Glut1DS), glucose transport into brain is reduced due to impaired Glut1 function in endothelial cells at the blood-brain barrier. This can lead to shortages of glucose in brain and is thought to contribute to seizures. Ketogenic diets are the first-line treatment and, among many beneficial effects, provide auxiliary fuel in the form of ketone bodies that are largely metabolized by neurons. However, Glut1 is also the main glucose transporter in astrocytes. Here, we review data indicating that glucose shortage may also impact astrocytes in addition to neurons and discuss the expected negative biochemical consequences of compromised astrocytic glucose transport for neurons. Based on these effects, auxiliary fuels are needed for both cell types and adding medium chain triglycerides (MCTs) to ketogenic diets is a biochemically superior treatment for Glut1DS compared to classical ketogenic diets. MCTs provide medium chain fatty acids (MCFAs), which are largely metabolized by astrocytes and not neurons. MCFAs supply energy and contribute carbons for glutamine and γ-aminobutyric acid synthesis, and decanoic acid can also block α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid glutamate receptors. MCTs do not compete with metabolism of ketone bodies mostly occurring in neurons. Triheptanoin, an anaplerotic but also gluconeogenic uneven MCT, may be another potential addition to ketogenic diets, although maintenance of "ketosis" can be difficult. Gene therapy has also targeted both endothelial cells and astrocytes. Other approaches to increase fuel delivery to the brain currently investigated include exchange of Glut1DS erythrocytes with healthy cells, infusion of lactate, and pharmacological improvement of glucose transport. In conclusion, although it remains difficult to assess impaired astrocytic energy metabolism invivo, astrocytic energy needs are most likely not met by ketogenic diets in Glut1DS. Thus, we propose prospective studies including monitoring of blood MCFA levels to find optimal doses for add-on MCT to ketogenic diets and assessing of short- and long-term outcomes.

A vegetable fat-based diet delays psychomotor and cognitive development compared with maternal dairy fat intake in infant gray mouse lemurs

Dairy fat has a unique lipid profile; it is rich in short- and medium-chain saturated fatty acids that induce ketone production and has a balanced ω6/ω3 ratio that promotes cognitive development in early life. Moreover, the high consumption of vegetable oils in pregnant and lactating women raises concerns regarding the quality of lipids provided to offspring. Here, we investigate maternal dairy fat intake during gestation and lactation in a highly valuable primate model for infant nutritional studies, the gray mouse lemur (Microcebus murinus). Two experimental diets are provided to gestant mouse lemurs: a dairy fat-based (DF) or vegetable fat-based diet (VF). The psychomotor performance of neonates is tested during their first 30 days. Across all tasks, we observe more successful neonates born to mothers fed a DF diet. A greater rate of falls is observed in 8-day-old VF neonates, which is associated with delayed psychomotor development. Our findings suggest the potential benefits of lipids originating from a lactovegetarian diet compared with those originating from a vegan diet for the psychomotor development of neonates.

Sustainable one-pot solvent-free enzymatic synthesis of capric acid-rich structured lipids to enhance the nutritional value of grape seed oil

Grape seed oil (GSO) is a valuable resource for the pharmaceutical, cosmetic, and food industries due to its fatty acid composition, which provides anti-inflammatory, cardioprotective, antimicrobial, and anticancer properties. GSO is also recognized for its sensory attributes and serves as an essential component in our dietary intake, providing vital fatty acids for metabolic reactions. The enzymatic synthesis of structured lipids (SL) using GSO was investigated in this study. The SL was created by incorporating medium chain fatty acids (capric acid – C10) at the sn-1 and sn-3 positions and long chain fatty acids in the internal position of the triacylglycerols (TAG). GSO was selected based on its composition rich in unsaturated fatty acids, mainly linoleic acid. The acidolysis reactions were catalyzed by an immobilized commercial lipase Lipozyme TLIM®. The incorporation degree of C10 into GSO reached 36%. Health lipid indices were calculated to determine the C10 incorporation effect on the improvement of nutritional quality of oil. The SL showed lower oxidative stability index (OSI) values than the original oil and higher degree of fatty acid (DFA) values. The simpler, solvent-free method not only enhances the health benefits of oils but also gives valuable properties to GSO, making it an even more attractive option for food industries.

Analysis of the intestinal microbiota and profiles of blood amino acids and acylcarnitines in neonates with hyperbilirubinemia

ObjectiveThis study aimed to discuss the distinctive features of the intestinal microbiota in neonates with hyperbilirubinemia and to comprehensively analyse the composition of the intestinal microbiota as well as the levels of free amino acids and acylcarnitines in the peripheral blood of neonates experiencing hyperbilirubinemia.ResultsAt the phylum level, Proteobacteria, Firmicutes, Actinobacteria, Bacteroidetes, and Chloroflexi were the five predominant microbial groups identified in both the hyperbilirubinemia and control groups. Alpha diversity analysis, encompassing seven indices, showed no statistically significant differences between the two groups. However, Beta diversity analysis revealed a significant difference in intestinal microbiota structure between the groups. Linear discriminant analysis effect size (LEfSe) indicated a significant reduction in the abundance of Gammaproteobacteria and Enterobacteriaceae within the hyperbilirubinemia group compared to that in the control group. The heatmap revealed that the control group exhibited increased abundances of Escherichia and Bifidobacterium, while the hyperbilirubinemia group exhibited increased levels of Enterococcus and Streptococcus. Regarding blood amino acids and acylcarnitines, there were greater concentrations of citrulline (Cit), arginine (Arg), ornithine (Orn), and valine (Val) in the hyperbilirubinemia group than in the control group. The hyperbilirubinemia group also exhibited significant increases in medium-chain fatty acids (C6, C8), long-chain fatty acids (C18), and free carnitine (C0).ConclusionBy comparing neonates with hyperbilirubinemia to those without, a significant disparity in the community structure of the intestinal microbiota was observed. The intestinal microbiota plays a crucial role in the bilirubin metabolism process. The intestinal microbiota of neonates with hyperbilirubinemia exhibited a certain degree of dysbiosis. The abundances of Bacteroides and Bifidobacterium were negatively correlated with the bilirubin concentration. Therefore, the fact that neonates with hyperbilirubinemia exhibit some variations in blood amino acid and acylcarnitine levels may provide, to a certain degree, a theoretical basis for clinical treatment and diagnosis.

Fabrication, characterization and application of Pickering emulsion gels stabilized by defatted grape seed powder

The feasibility of defatted grape seed powder (DGSP) stabilizing Pickering emulsion gels as butter substitute was investigated. The Pickering emulsion gel was constructed using DGSP through high-speed homogenization, and the effects of particle concentration (c) and oil-phase (Medium chain triglyceride) volume fraction (φ) on its structure and properties were investigated. Its application as a butter substitute was also evaluated. The results showed that DGSP had various particle shapes, a wide particle size distribution (3–130 μm), and a three-phase contact angle of 128.9 ± 2.3°. The O/W Pickering emulsion gels with φ ≥ 60% could be obtained at c ≥ 2%. The droplet diameter was negatively correlated with c and positively correlated with φ, while the gel strength was positively related to c and φ. The resulting emulsion gel demonstrated solid-like viscoelastic behavior and pseudoplasticity, and had the potential to serve as a butter substitute. The results can promote the application of grape seeds in food.

Analysis of dietary fats intake and lipid profile in Chilean patients with glucose transport type 1 deficiency syndrome: similarities and differences with the reviewed literature.

Glucose transporter type 1 deficiency syndrome (GLUT1-DS) is a neurological disorder caused by mutations in the SLC2A1 gene. The main treatment is ketogenic diet therapy (KDT), which changes the brain's energy substrate from glucose to ketone bodies. The diet controls seizures, but there may be side effects such as dyslipidemia. This study aimed to describe the type of fats ingested by the Chilean cohort of patients with GLUT1-DS and analyze for alterations in the lipid profile. A GLUT1-DS group and a control group were formed, each with 13 subjects who were matched by age, gender, and nutritional status. Anthropometry, dietary intake, including types of fat, and blood tests were evaluated (lipid and liver profile, and 25-hydroxyvitamin D levels). A high-fat diet, especially saturated fat, was identified in the GLUT1-DS group (38% of total calories), with the use of medium-chain triglycerides (17% of total calories). In addition, GLUT1-DS participants had a higher intake of monounsaturated (MUFA) and polyunsaturated (PUFA) fats and adequate consumption of omega-3 (2% of total calories). Despite the GLUT1-DS group receiving on average 80% of its total energy as fats, it is important to highlight that 50% are MUFA+PUFA fats, there were no significant differences in the lipid and liver profile compared to the control group. KDT did not negatively impact lipid profile, despite a high intake of fats. It is important to monitor lipid profiles, in a personalized and constant manner, to prevent future nutritional risks.

Impact of MCT Oil and Caffeine on Substrate Metabolism during Submaximal Exercise

Introduction: Recent research has suggested that medium-chain triglyceride (MCT) supplementation may increase fat oxidation (FatOx) during aerobic exercise, sparing muscle glycogen and, perhaps, enhancing performance. As both MCT and caffeine (CAF) are theorized to elicit these effects, this pilot study’s purpose was to compare the physiological responses of their combined supplementation during submaximal cycling exercise. Methods: Eight aerobically trained males (mean±SD; age 23.6±4.4 years; body mass 82.3±15.8 kg; height 180.9±8.7 cm) completed one aerobic capacity (VO2peak) test and three 45-min exercise trials at 60% VO2peak. Blinded and counterbalanced, one-hour prior to each trial, participants consumed: MCT+CAF (20 mL + 100 mg), long-chain triglycerides (LCT)+CAF (40 mL + 100 mg), or CAF (100 mg). Oxygen consumption (VO2), carbon dioxide output (VCO2), and minute ventilation (VE) were measured, subsequently calculating respiratory exchange ratio (RER), energy expenditure (EE), carbohydrate oxidation (CarbOx), and FatOx. Results: No significant differences between conditions were observed for average VO2 (p=0.474; η2=0.101), VE (p=0.323; η2=0.149), (RER (p=0.323; η2=0.149), EE (p=0.474; η2=0.101), CarbOx (p=0.274; η2=0.169), or FatOx (p=0.478; η2=0.100) or for total EE (p=0.474; η2=0.101), CarbOx (p=0.274; η2=0.169), or FatOx (p=0.478; η2=0.100). Conclusions: Co-ingestion of MCT+CAF didn’t produce any significantly different physiological responses compared to co-ingestion of LCT+CAF or the CAF control.

The Effect of Acid Hydrolysis on the Pickering Emulsifying Capacity of Tartary Buckwheat Flour.

The effect of sulfuric acid hydrolysis on the Pickering emulsifying capacity of Tartary buckwheat flour (TBF) rich in starch was evaluated for the first time. The results indicate that the sulfuric acid concentration and hydrolysis time had a significant impact on the Pickering emulsifying capacity of acid-hydrolyzed Tartary buckwheat flour (HTBF). A low sulfuric acid concentration (1-2 mol/L) could reduce the particle size of HTBF, but it also decreased the Pickering emulsifying ability. At a sulfuric acid concentration of 3 mol/L, appropriate treatment time (2 and 3 days) led to particle aggregation but significantly improved wettability, thereby resulting in a rapid enhancement in emulsifying capacity. Under these conditions, the obtained HTBF (HTBF-D2-C3 and HTBF-D3-C3) could stabilize medium-chain triglyceride (MCT)-based Pickering high-internal-phase emulsions (HIPEs) with an oil-phase volume fraction of 80% at the addition amounts (c) of ≥1.0% and ≥1.5%, respectively. Its performance was significantly superior to that of TBF (c ≥ 2.0%). Furthermore, at the same addition amount, the droplet size of HIPEs constructed by HTBF-D3-C3 was smaller than that of HTBF-D2-C3, and its gel strength and microrheological performance were also superior to those of HTBF-D2-C3, which was attributed to the higher wettability of HTBF-D3-C3. The findings of this study can facilitate the in-depth application of Tartary buckwheat and provide references for the development of novel Pickering emulsifiers.

Exogenous Ketone Supplement Administration Abrogated Isoflurane-Anesthesia-Induced Increase in Blood Glucose Level in Female WAG/Rij Rats.

It has been demonstrated that isoflurane-induced anesthesia can increase the blood glucose level, leading to hyperglycemia and several adverse effects. The administration of a mix of ketone diester (KE) and medium-chain triglyceride (MCT) oil, named KEMCT, abolished the isoflurane-anesthesia-induced increase in blood glucose level and prolonged the recovery time from isoflurane anesthesia in a male preclinical rodent model, Wistar Albino Glaxo/Rijswijk (WAG/Rij) rats. While most preclinical studies use exclusively male animals, our previous study on blood glucose changes in response to KEMCT administration showed that the results can be sex-dependent. Thus, in this study, we investigated female WAG/Rij rats, whether KEMCT gavage (3 g/kg/day for 7 days) can change the isoflurane (3%)-anesthesia-induced increase in blood glucose level and the recovery time from isoflurane-evoked anesthesia using the righting reflex. Moreover, KEMCT-induced ketosis may enhance both the extracellular level of adenosine and the activity of adenosine A1 receptors (A1Rs). To obtain information on the putative A1R mechanism of action, the effects of an A1R antagonist, DPCPX (1,3-dipropyl-8-cyclopentylxanthine; intraperitoneal/i.p. 0.2 mg/kg), on KEMCT-generated influences were also investigated. Our results show that KEMCT supplementation abolished the isoflurane-anesthesia-induced increase in blood glucose level, and this was abrogated by the co-administration of DPCPX. Nevertheless, KEMCT gavage did not change the recovery time from isoflurane-induced anesthesia. We can conclude that intragastric gavage of exogenous ketone supplements (EKSs), such as KEMCT, can abolish the isoflurane-anesthesia-induced increase in blood glucose level in both sexes likely through A1Rs in WAG/Rij rats, while recovery time was not affected in females, unlike in males. These results suggest that the administration of EKSs as an adjuvant therapy may be effective in mitigating metabolic side effects of isoflurane, such as hyperglycemia, in both sexes.

Fatty acids metabolism in ozone-induced pulmonary inflammatory injury: Evidence, mechanism and prevention

Ozone (O3) is a major air pollutant that directly threatens the respiratory system, lung fatty acid metabolism disorder is an important molecular event in pulmonary inflammatory diseases. Liver kinase B1 (LKB1) and nucleotide-binding domain leucine-rich repeat-containing protein 3 (NLRP3) inflammasome not only regulate inflammation, but also have close relationship with fatty acid metabolism. However, the role and mechanism of LKB1 and NLRP3 inflammasome in lung fatty acid metabolism, which may contribute to ozone-induced lung inflammation, remain unclear, and effective strategy for preventing O3-induced pulmonary inflammatory injury is lacking. To explore these, mice were exposed to 1.00 ppm O3 (3 h/d, 5 days), and pulmonary inflammation was determined by airway hyperresponsiveness, histopathological examination, total cells and cytokines in bronchoalveolar lavage fluid (BALF). Targeted fatty acids metabolomics was used to detect medium and long fatty acid in lung tissue. Then, using LKB1-overexpressing adenovirus and NLRP3 knockout (NLRP3−/−) mice to explore the mechanism of O3-induced lung fatty acid metabolism disorder. Results demonstrated that O3 exposure caused pulmonary inflammatory injury and lung medium and long chain fatty acids metabolism disorder, especially decreased dihomo-γ-linolenic acid (DGLA). Meanwhile, LKB1 expression was decreased, and NLRP3 inflammasome was activated in lung of mice after O3 exposure. Additionally, LKB1 overexpression alleviated O3-induced lung inflammation and inhibited the activation of NLRP3 inflammasome. And we found that pulmonary fatty acid metabolism disorder was ameliorated of NLRP3 −/− mice compared with those in wide type mice after O3 exposure. Furthermore, administrating DGLA intratracheally prior to O3 exposure significantly attenuated O3-induced pulmonary inflammatory injury. Taken together, these findings suggest that fatty acids metabolism disorder is involved in O3-induced pulmonary inflammation, which is regulated by LKB1-mediated NLRP3 pathway, DGLA supplement could be a useful preventive strategy to ameliorate ozone-associated lung inflammatory injury.

Combining lipase enzymatic techniques and antioxidants on the flavor of structured lipids (SLs) prepared from goat butter and coconut oil

Enzymatic transesterification can improve the quality of goat butter, however, the high temperature may lead to oil oxidation and produce a bad taste. In this study, goat butter (GB) and coconut oil (CO) were combined to synthesize structured lipids (SLs) by enzymatic transesterification, and antioxidants were added to inhibit oxidation to improve the oils’ flavor. The Lipozyme TLIM and tert-butylhydroquinone (TBHQ) were screened for SLs synthesis. The contents of SLs were 56.24% under the optimal conditions of 8 h reaction time, 5°C reaction temperature, 8% enzyme dosage, and 3:1 (w/w) substrate ratio, and the addition of antioxidants had no significant effect on the contents of SLs. It was demonstrated that short-chain fatty acids in the SLs were significantly reduced by 59.2%, and the medium-chain fatty acids were increased by 54.9% compared to GB. One hundred volatile compounds were identified in the lipid samples and classified as (1) long-chain alkane aromatic components and aromatic components, which remarkably decreased by 58.9%, and (2) alcohols, aldehydes, and ketones, which increased by 12.9%, through using solid-phase microextraction coupled with gas chromatography-mass spectrometry (SPME-GC-MS) and electronic nose. The relative aroma activity value (ROAV) of 9 key volatile aroma components was higher than 1 in the SLs, which greatly contributed to the reduction of goaty flavor and increasing cream flavor. The results suggested that the goaty flavor of GB could be significantly reduced by decreasing the short-chain fatty acids contents in SLs through enzymatic transesterification with CO, while the addition of antioxidants efficiently inhibited excessive lipids oxidation and increased the content of long-chain aldehydes, ketones, and lactones, thus enhancing the cream flavor, which had great potential to be utilized as natural food flavor additives.

The Effect of Virgin Coconut Oil on TNFα and Cytochrome P450 Aromatase Levels in Obese Female White Rats (Rattus norvegicus) Induced with a High-Fat Diet

Background: The high level of various proinflammatory cytokines due to adipocyte hypertrophy such as TNFα in obesity increases cytochrome-P450-aromatase, further inducing increased estrogen production, hypersecretion of Luteinizing Hormone and decreased Follicle Stimulating Hormone, resulting in impaired folliculogenesis, and chronic anovulation. Virgin Coconut Oil is rich in Medium Chain Fatty Acid and phytochemicals as anti-inflammatory. Aim: The aim of this study was to determine the effect of VCO on reducing TNFα and cytochrome-P450-aromatase levels in obese female white rats. Methods: Post test only control group, 24 rats were divided into 3 groups, namely: negative control group (K-), positive control group (K+), and treatment group (P). The K+ and P groups were induced using a high-fat diet for 10 weeks, then the P group was given VCO in weeks 7-10. On the first day of week 11, blood was taken from the orbital vein of the eyes of all rats for TNFα levels. Both examinations used ELISA kits. Data analysis used Shapiro Wilks normality test. The value of p>0.05, followed by hypothesis testing using one-way annova and post hoc Bonferroni. Results: Mean TNFα in the K- group: 141.71±8.87, K+ group: 15.09±19.30, P group: 162,09±13,57. Mean cytochrome-P450-aromatase levels in the K- group: 9.00±0.77, K+ group: 12.22±1.69, P group: 10.58±0.97. The mean levels of one way ANOVA analysis showed that the administration of VCO can significantly reduce TNFα levels (p=0.001) and significantly reduce Cytochrome-P450-aromatase levels (p=0.001). Conclusion: VCO administration can reduce TNFα levels and cytochorme-P450-aromatase levels in obese female white rats. Keywords: TNFα, cytochrome-P450-aromatase, infertility, obesity

Diaminated Cellulose Beads as a Sustainable Support for Industrially Relevant Lipases.

Environmentally persistent polystyrene or polyacrylic beads are used as supports in enzyme large-scale bioprocesses, including conversion glucose isomerization for high-fructose corn syrup production, hydrolysis of lactose, and synthesis of active pharmaceutical ingredients. In this paper, we report the development of a novel sustainable and scalable method to produce diaminated cellulose beads (DAB) as highly efficient alternative supports for industrially relevant lipases. Regenerated cellulose beads were grafted with diaminated aliphatic hydrocarbons via periodate oxidation and reductive amination. The oxidation step indicated that aldehyde content can be easily tuned through the reaction time and concentration of reactants. Reductive amination of dialdehyde cellulose was more efficient as the length of the diaminated hydrocarbon compound increased. Morphological analysis of DAB showed that cellulose chemical grafting enabled the preservation of the bead shape and internal structure upon freeze-drying. Enzymatic degradability studies demonstrated that chemical functionalization did not undermine enzyme cellulose hydrolysis. The addition of aminated moieties on cellulose dramatically increased absorption efficiency for all industrially relevant lipases used, reaching 100% for Thermomyces lanuginosus lipase (TLL). Storage and recyclability experiments demonstrated that enzymes were retained and recyclable for at least nine cycles, although the activity gradually declined after each cycle. Medium chain triacylglycerol hydrolysis in a SpinChem reactor using TLL immobilized on 1,6 DAB exhibited higher activity compared to acrylic beads (588 vs 459 U/g) suggesting that biodegradable cellulose-based materials could be a valid and attractive alternative to plastics carriers.

PSIII-2 Evaluation of medium-chain fatty acids on just-weaned piglet performance and diarrhea scores: A nursery trial

Abstract Weaning is one of the most stressful events for commercial pigs. The transition from mother’s milk (animal proteins) to a solid diet based on plant proteins, the reduction in lactose intake and the decline in HCl secretion by the digestive system of the piglet result in a sudden and significant increase in stomach pH as well as an increased risk of post-weaning diarrhea. The aim of this trial was to evaluate whether the use of a blend of medium-chain fatty acids (MCFAs) on top-off conventional nursery diets would improve zootechnical performance and intestinal problems in just-weaned piglets. Piglets [n = 624, 21-d of age, 6.5 kg body weight (BW), (♂ Fast Duroc x ♀ Fast 275)] were housed in 26 pens of an experimental nursery. Each pen (24 pigs/pen, with a starting density of 3.0 ft2/pig) was equipped with a height-adjustable water sucker, a water bowl and fitted with a single feeder-bin. Females and males were allocated to each pen, according to 3 BW categories (heavy, medium, and light). During a 46-d period of trial, the piglets (13 replicated pens per treatment) were allotted to a conventional 5 phases feeding program with 2 kg/MT of chlortetracycline (22%) and 400 g/MT of tiamuline (1.78%; CON). These basal diets contained 2,500 ppm Zn in Phases #1, 2, and only 2,000 ppm Zn in phase #3. Phases #1 to 3 were also supplemented with Ca-butyrate (1 kg/MT) and a blend of protected acidifiers (1 kg/MT). The treated group consisted to the basal diet (CON) supplemented with a blend of medium-chain fatty acids (MCFAs) at 1.0 or 0.75 kg/MT for Phases #1, 2,3 and Phases #4, 5, respectively and supplying of a proprietary blend of caprylic, capric, and lauric acids. Daily feed consumption, weekly pig pen BW, dead and removed pigs and daily diarrhea evaluation index were completed. Data are analyzed by an ANOVA model including treatment and pen as fixed and random effects, respectively. Pairwise comparisons were completed with a Chi-square test. The overall results are presented in Table 1. At the end of wk 2 and 3, animals receiving MCFAs had a better average daily gain (ADG; P < 0.05) than CON. Therefore, at wk 2, 3 and 4, MCFAs animals BW were heavier than CON ones (P < 0.05). Average daily feed intake (ADFI) was not significantly different for the overall duration of the trial (P > 0.05). However, at weeks 2 and 3, MCFAs piglets had a greater ADFI than CON ones (P < 0.05). The daily diarrhea index of the MCFAs group was less than CON at wk 3 (P < 0.05). In this trial, the significant improvement of diarrhea index on just-weaned MCFAs treated piglets could explain the significant better growth during the first 3 wk of nursery.

PSV-4 Exposure to prenatal tributyrin improved pre-weaning growth performance and reduced fat deposition in the muscle of market age pigs

Abstract Organic acids, including short chain fatty acids, medium chain fatty acids, and tricarboxylic acid, are used as alternatives to antibiotics for the purpose of growth promotion in livestock species. Butyric acid has been used extensively for its various functions in intestinal health and barrier function, as well as its actions as a histone deacetylase inhibitor. Many of the issues relating to using butyric acid as a feed supplement are overcome by utilizing tributyrin (TB), a butyrate prodrug. Dietary supplementation of TB to sows during gestation was able to improve pre-weaning litter performance. Sows (n = 52, 42 gestational days) were randomly assigned to either a control diet or control diet supplemented with 2% TB. All diets were made isocaloric with the addition of oil and wheat flour. Dietary treatments continued until 112 d of gestation. After farrowing, piglet growth performance was monitored. Once offspring reached 6 mo of age, 30 gilts from the most median performing litters were identified for tissue collection and evaluation of meat characteristics. Sows that received tributyrin during gestation tended to have greater total litter weights at birth (P < 0.1); however, average individual birthweights were not statistically different between control and TB litters. The TB piglets tended to be heavier at weaning (P < 0.1). Pre-weaning average daily gains were higher in piglets exposed to prenatal TB (P < 0.01). Body weights and hot carcass weights did not differ significantly in gilts at market age. There was, however, a trend of increased dressing percentage from TB treated offspring, accompanied by significantly reduced back fat at the 10th rib (P < 0.1 and P < 0.05 respectively). These data suggest that TB supplementation results in both temporary and permanent improvements to growth and development. This study provides further evidence that prenatal TB can have long term effects on muscle growth and meat quality. Further studies are needed to ultimately determine whether prenatal or postnatal supplementation is ideal for improving long-term growth performance.

PSV-18 Improved performance of PRRSV challenged pigs fed 1-α-glycerol monolaurate

Abstract Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and Porcine Enteric Diarrhea (PEDV) has posed a persistent challenge to the swine industry despite vaccinations and biosecurity measures. Feed has been shown to be a carrier for bacterial and viral pathogens (Dee et al., 2020). Feed mitigation using medium chain fatty acids can eliminate the pathogen or reduce the pathogen load below its infectious dose. However, less is known regarding the application of monoglycerides for feed mitigation. The objective of this study was to determine if using only 1-α-glycerol monolaurate (GML) can be an effective feed mitigant against PRRSV compared with untreated control feed. A total of 2 rooms, each with 100 pigs per room, and 6 pens per room were used. The initial body weight of the pigs averaged 15 kg and were sourced from a herd free of PRRSV and PEDV. GML was added to the feed at 2 kg/mT. The study period was 14 d with starting and end body weights measured. An ice block viral challenge as described by Dee et al. (2020) was used. Briefly, a 500 mL ice cube containing 1x107 50% tissue culture infectious dose of each virus [PRRSV, PEDV and Senecavirus A (SVA)] was added to 1,300 kg of feed on d 0 and again on d 6. On d 15 pigs were tested for PRRSV (serum), PEDV (rectal swabs) and SVA (tonsil samples). Control and GML fed pigs did not seroconvert to PEDV and SVA, therefore, only results for PRRSV are presented. Average daily gain was significantly improved in GML fed pigs (0.50 kg/d vs 0.20 kg/d; P < 0.05). Percentage of PRRSV positive feed samples were decreased (P < 0.05) in GML-treated feed from 100% (Ct = 32.2) in control feed to 33% (Ct = 34.3). The percentage of positive oral fluid samples decreased (P < 0.05) from 100% (Ct = 26.1) in control pigs to 17.0% (Ct = 33.0) in GML fed pigs. GML fed pigs exhibited no clinical signs of PRRS (dysponea), whereas 100% of the control pigs had clinical signs (P < 0.05). Similarly, GML fed pigs had no PRRSV positive serum samples but 100% of the control pigs had PRRSV positive serum samples. Results from this study show that the use of only GML can be an effective feed mitigant to reduce or eliminant the clinical signs of PRRS in pigs and maintain growth performance.

308 Awardee Talk: Prevention and mitigation of pathogens in feed and feed manufacturing systems

Abstract Prior to 2013, pathogen transmission through feed was focused primarily on bacteria and biosecurity was used to prevent adulterant entry into the ingredients and the feed mills. Following the porcine epidemic diarrhea virus (PEDV) outbreak in 2013, research focused on pathogen mitigation and control quickly became a subject of interest. Since PEDV, multiple endemic viruses and foreign animal diseases have been found to be transmissible in feed or feed ingredients. As African swine fever virus (ASFV) spread closer to the United States and endemic pathogens are a continued pressure on production systems, it is imperative that prevention measures are included in the feed supply chain and options for mitigation are fully elucidated for times of need. Previous research has demonstrated that preventative measures such as boot baths, lines of separation, showers, and changing clothes are effective measures when appropriately maintained. However, these protocols can seem costly and burdensome to employees, especially when the risk of disease transmission appears low. Employee training is difficult as disease transmission is difficult to visualize and exhibit the importance of each measure. Glo Germ, a florescent substance, is a cost-effective training tool which can demonstrate the efficacy of prevention protocols and can identify areas where biosecurity measures have not been properly utilized. Preventative measures are the first line of defense for feed manufacturing systems, but understanding and using mitigation techniques in the event of contamination can limit pathogen spread and protect the rest of the feed supply chain. Pelleting, as a method of thermal inactivation process, has recently been demonstrated to lessen the quantity of detectable viral RNA and decrease the risk of infection. The pelleting process, however, is a point-in-time mitigant and does not decrease the risk of product re-contamination as feed continues down the supply chain. The use of chemical mitigants, like formaldehyde, medium-chain fatty acids, and organic acids, can decrease the presence of bacterial and viral pathogens with longer-lasting mitigation properties. These mitigants can either be used in the feed itself, or as an addition to flushing batches which help disperse the chemical mitigants throughout the feed manufacturing equipment, thus reducing the risk of product re-contamination. In the event more drastic measures must be taken to eliminate pathogen presence from feed mills, gaseous fumigation and the use of portable heaters show promise as methods to decontaminate feed manufacturing equipment without the use of water or complete equipment breakdown. Overall, effective training and implementation of preventative measures are needed to provide pathogen-free feed to production systems, but continued research in mitigation and mill decontamination will further protect the feed supply chain in the event of pathogen contamination.