Dietary yeast glycoprotein supplementation improves the growth performance, intestinal health and disease resistance of largemouth bass (Micropterus salmoides) fed low-fishmeal diets.

The active ingredients extracted from yeast are important for regulating animal health. The aim of the current research was to explore the impacts of dietary yeast glycoprotein (YG) on the growth performance, intestinal morphology, antioxidant capacity, immunity and disease resistance of largemouth bass (Micropterus salmoides). A total of 375 juvenile fish (6.00 ± 0.03 g) were allocated into 15 fiberglass tanks. Triplicate tanks were assigned to each diet. The dietary YG inclusion was as follows: the first group was given a high fishmeal diet (40% fishmeal, 0% YG) (FM) and the second group was given a low fishmeal diet (30% fishmeal and 15% soybean meal, 0% YG) (LFM). The fish in the third, fourth and fifth groups were fed the LFM diet supplemented with 0.5% (LFM+YG0.5), 1.0% (LFM+YG1.0) and 2.0% (LFM+YG2.0) YG, respectively. After a 60- day feeding trial, a challenge test using A. hydrophila was carried out. The results showed that the final body weight (FBW) and weight gain rate (WGR) in the LFM+YG2.0 group were significantly higher than those in the LFM group and were no significantly different from those in the FM group. This may be partially related to the activation of the target of rapamycin (TOR) signaling pathway. Dietary YG supplementation enhanced intestinal physical barriers by upregulating the intestinal tight junction protein related genes (claudin1, occludin and zo2) and improving the structural integrity of the gut, which may be partially associated with AMPK signaling pathway. Moreover, dietary YG increased the antioxidant capacity in the gut, upregulated intestinal anti-inflammatory factors (il-10, il1-1ß and tgf-ß) and downregulated proinflammatory factors (il-1ß and il-8), which may be partially related to the Nrf2/Keap1 signaling pathways. The results of the challenge test indicated that dietary supplementation with 0.5 or 1.0% YG can increase the disease tolerance of largemouth bass against A. hydrophila. In conclusion, the present results indicated that dietary supplementation with YG promotes the growth performance, intestinal immunity, physical barriers and antioxidant capacity of largemouth bass. In addition, 1.0% of dietary YG is recommended for largemouth bass based on the present results.

Antioxidant and prooxidant activities of tea polyphenols in oil-in-water emulsions depend on the level used and the location of proteins.

We studied the impacts of protein location (interface or aqueous phase) on the antioxidant and prooxidant activities of tea polyphenols (TP) in model oil-in-water emulsions (pH 7) at a low (0.01% w/v) or high (0.04 % w/v) concentration. TP at 0.01% reduced the levels of both lipid and protein oxidation markers in emulsions, independent of the protein location. However, TP were more potent when proteins were located at the interface. At 0.04%, TP were only weakly antioxidant towards lipids but were prooxidant towards proteins in emulsions with proteins at the interface, whereas they were still somewhat antioxidant for aqueous phase proteins. These results indicate that TP may act as either antioxidants or prooxidants depending on their concentration and also on the location of the proteins in emulsions. The level of TP should be optimized for emulsion-based foods or beverages to achieve optimum antioxidant activity.

Formation of Antioxidant Multilayered Coatings for the Prevention of Lipid and Protein Oxidation in Oil-in-Water Emulsions: Lycium barbarum Polysaccharides and Whey Proteins.

The impact of Lycium barbarum polysaccharides (LBPs) on the physical and chemical stability of oil-in-water emulsions coated by a whey protein isolate (WPI) was investigated. At pH 3.0, the anionic LBP (0.2-0.6 wt %) molecules were electrostatically deposited onto the cationic surfaces of the WPI-coated oil droplets, leading to the formation of stable multilayered emulsions containing WPI-/LBP-coated oil droplets. However, increasing the LBP concentration to 0.8 wt % led to oil droplet aggregation, which was attributed to charge neutralization, bridging flocculation, and/or depletion flocculation. For subsequent experiments, a low (0.2%) and an intermediate (0.6%) LBP dose was used to prepare the secondary emulsions, and then their physical and oxidative stability was studied during 8 days of storage at 37 °C. The presence of the multilayer WPI/LBP coatings around the oil droplets inhibited lipid oxidation (reduced levels of lipid hydroperoxides and 2-thiobarbituric acid-reactive substances), as well as protein oxidation (reduced levels of carbonyl formation, sulfhydryl consumption, molecular weight modifications, intrinsic fluorescence loss, and Schiff-base fluorescence gain). The antioxidant effects of the multilayer coatings were greater at the higher LBP concentration. These results suggest that LBP, a natural plant-based polysaccharide isolated from a traditional Chinese medicine, can be used to improve the quality of emulsion-based foods. However, the level used should be optimized to ensure good physical and oxidative stability of the emulsions.

Impact of tea polyphenols on the stability of oil-in-water emulsions coated by whey proteins.

The ability of tea polyphenols (0, 0.01, 0.02 or 0.04 w/v %) to inhibit lipid and protein oxidation in walnut oil-in-water (O/W) emulsions was examined, as well as to alter their stability to aggregation and creaming. The lipid droplets in these emulsions were coated by whey proteins. The physical stability of the emulsions during storage (50 °C, 96 h) was improved by addition of 0.01% tea polyphenols, but reduced when higher levels were added. Low levels (0.01%) of tea polyphenols inhibited lipid oxidation (lipid hydroperoxide and 2-thiobarbituric acid-reactive substance formation) and protein oxidation (carbonyl and Schiff base formation, sulfhydryl and intrinsic fluorescence loss, and molecular weight changes). However, high levels (0.04%) of tea polyphenols were less effective at inhibiting lipid oxidation, and actually promoted protein oxidation. Tea polyphenols are natural antioxidants that can enhance the quality and shelf life of emulsified polyunsaturated lipids when used at an appropriate concentration.

Inhibition of Lipid and Protein Oxidation in Whey-Protein-Stabilized Emulsions Using a Natural Antioxidant: Black Rice Anthocyanins.

The food industry is exploring the natural environment to identify botanical extracts that can be used as functional ingredients that can replace synthetic ingredients in foods. In the present study, the ability of black rice anthocyanins as natural antioxidants to inhibit both lipid and protein oxidation in protein-stabilized oil-in-water emulsions was examined. Whey-protein-stabilized emulsions were prepared containing 0, 0.02, 0.04, and 0.06% (w/v) anthocyanins, and then the impact of this plant-based extract on their physical and chemical stabilities was evaluated. The addition of the anthocyanins improved the physical stability of the emulsions in a dose-dependent manner by inhibiting droplet aggregation during storage (35 °C for 5 days). The anthocyanins also exhibited good antioxidant activity in a dose-dependent manner, as seen by their capacity for inhibiting both lipid oxidation (reduced lipid hydroperoxides and malondialdehyde) and protein oxidation (reduced carbonyl and Schiff base formation, intrinsic fluorescence loss, and molecular weight changes). Black rice anthocyanins may therefore be an effective botanical extract for improving the stability of protein-stabilized food emulsions by inhibiting oxidative reactions.

A potential alternative to traditional antibiotics in aquaculture: Yeast glycoprotein exhibits antimicrobial effect in vivo and in vitro on Aeromonas caviae isolated from Carassius auratus gibelio.

In aquaculture, antibiotics are commonly used to provide protection against pathogens; however, this practice has become controversial due to increased occurrences of microbial resistance, and alternatives are needed. This study aimed to investigate the antimicrobial activity of yeast glycoprotein (YG) against Aeromonas caviae. Pathogens were isolated from liver of diseased Carassius auratus gibelio. Based on morphological and biochemical analysis, together with 16S rRNA gene sequencing, the isolated strains were identified as A. caviae and concluded as clones of a single strain and named L2. Further pathogenicity analysis revealed that A. caviae possessed ß-haemolysis, and its median lethal dose for C. gibelio was 1.33 × 106  CFU/ml. Hepatic adenylate kinase and pyruvate kinase activities of C. gibelio were inhibited post-A. caviae infection. Antimicrobial drug test suggested that A. caviae was a multidrug-resistant organism but could be inhibited by YG in vitro. Minimum inhibitory and bactericidal concentration of YG was 83.3 mg/ml and 166.7 mg/ml, respectively. Microbiota sequencing results showed that YG supplement could obviously decrease the relative abundance of Aeromonas and increase the microbial diversity. Our study revealed that A. caviae from C. gibelio was a multidrug-resistant bacteria strain, and could be significantly inhibited by YG in vivo and in vitro, thus providing important insights into ecological control and pathogenesis of A. caviae in aquaculture.

Effects of dietary yeast hydrolysate on the growth, antioxidant response, immune response and disease resistance of largemouth bass (Micropterus salmoides).

A 56-day growth trial was conducted to investigate the effects of dietary yeast hydrolysate on the growth performance, antioxidation, immune response and resistance against Aeromonas hydrophila in largemouth bass. Four experimental diets were prepared with yeast hydrolysate levels of 0% (Y0), 1.5% (Y1.5), 3.0% (Y3.0) and 4.5% (Y4.5). Each diet was randomly assigned to triplicate 150-L tanks and each tank was stocked with 30 largemouth bass (initial body weight, IBW = 7.71 ±â€¯0.02 g). A challenge test was carried out after the feeding trial by injecting A. hydrophila intraperitoneally for 4-day observation. The results showed that the FBW and WGR in Y1.5 group were significantly higher than those in Y0 group (P < 0.05) and the feed conversion ratio (FCR) got the lowest value in Y1.5 group. And the hydrolysate supplement significantly increased the 4-day cumulative survival rate after the bacterial challenge (P < 0.05). The plasma malondialdehyde was lower in the yeast hydrolysate supplement groups in both pre- and post-challenge test (P < 0.05), while the plasma C3 increased (P < 0.05). In post-challenge test, the plasma superoxide dismutase (SOD) and catalase (CAT) activities increased in the Y1.5 and Y3.0 groups respectively (P < 0.05), and plasma lysozyme in Y1.5 group and the plasma IgM in Y3.0 group were higher than those in others respectively (P < 0.05). For the q-PCR results, in post-challenge test, the hepatic hep2 expression level in Y1.5 and Y4.5 groups were both significantly higher than those in others (P < 0.05), as well as il-8 in Y3.0 group. The spleen hif-1alpha and tgf-beta1 expression levels in Y4.5 group were all significantly lower than those in others (P < 0.05), while the gilt was significantly higher (P < 0.05) in the post-challenge test. And the expression levels of spleen tnf-alpah1 in Y1.5 and Y3.0 groups and il-8 in Y3.0 group were all significantly higher than those in other groups (P < 0.05) in the post-challenge test. The head kidney gilt expression level was significantly higher in the yeast hydrolysate supplement groups compared with the Y0 group (P < 0.05), and the head kidney il-8 expression level in Y1.5 group was significant higher than those in other groups in post-challenge test (P < 0.05). The present results indicated dietary yeast hydrolysate improved the antioxidant ability and enhanced the immune response of largemouth bass without negative effect on growth. And 1.5% or 3.0% of dietary yeast hydrolysate was recommended for largemouth bass based on the present results.

Comparison of natural and synthetic surfactants at forming and stabilizing nanoemulsions: Tea saponin, Quillaja saponin, and Tween 80.

HYPOTHESIS: This study compared the interfacial and emulsification properties of tea saponins, quillaja saponins, and Tween 80. We hypothesized that tea saponins are an effective and sustainable source of plant-based emulsifiers that could replace synthetic or animal-based emulsifiers in many commercial applications. EXPERIMENTS: Interfacial tension measurements were used to characterize the behavior of the three surfactants at an oil-water interface. The emulsifying properties of the surfactants were determined by preparing oil-in-water emulsions containing 10 wt% medium chain triglycerides (MCT) and varying surfactant levels (0.1-2 wt%) using high-pressure homogenization (pH 7). The impact of surfactant type on emulsion formation and stability was determined by measuring particle size, zeta-potential, microstructure, and creaming stability. FINDINGS: The tea saponins were capable of producing nano-scale droplets (d32 < 200 nm) at low surfactant-to-oil ratios (SOR < 0.1). Emulsions containing tea saponins remained stable to droplet aggregation when exposed to various temperatures (30-90 °C), salt levels (0-200 mM NaCl), and pH values (3-9). However, droplet flocculation and/or coalescence occurred under highly acidic (pH 2) and high ionic strength (300-500 mM NaCl) conditions. Tea saponin-coated oil droplets appeared to be mainly stabilized by a combination of electrostatic and steric repulsion. The tea saponins behaved similarly or better than the other two emulsifiers under most conditions. These results suggest that tea saponins are effective plant-based surfactants that may have applications in commercial products.

The effects of long-term low selenium diet on the expression of CHST-3, CHST-12 and UST in knee cartilage of growing rats.

OBJECTIVES: To investigate the effect of low selenium diet on rat´s knee cartilage and expression of chondroitin sulfate (CS) sulfated enzymes in articular and epiphyseal-plate cartilage of rats' femur and tibia. METHODS: Twenty-four SD rats were randomly divided into two groups with six female and six male in each group: control group (selenium 0.18 mg/kg), and low selenium group (selenium 0.02 mg/kg). After 109 days, the rats were sacrificed. The ultrastructural changes in chondrocytes of rat knee cartilage were observed by transmission electron microscopy (TEM). The morphology and pathology changes of knee cartilage were examined by hematoxylin-eosin (HE) and toluidine blue (TB) staining. The localization and expression of enzymes involved in CS sulfation, including chondroitin 6-O-sulfotransferase 1 (CHST-3), chondroitin 4-O-sulfotransferase 2 (CHST-12) and uronyl 2-O-sulfotransferase (UST) were examined by immunohistochemical staining and semi-quantitative analysis. RESULTS: In low selenium group, ultrastructural changes of chondrocytes were observed in articular cartilage of femur (AF), articular cartilage of tibia (AT), epiphyseal-plate cartilage of femur (EF) and epiphyseal-plate cartilage of tibia (ET); however, no significant changes in chondrocytes number were observed in the above AF, AT, EF or ET. Moreover, reduced thickness of cartilage layer in AF, EF and ET was detected along with reduced staining areas of sulfated glycosaminoglycan in EF and ET in low selenium group. In addition, positive staining rate of CHST-3 was lower in AF, AT and EF, while positive staining rates of CHST-12 and UST were lower in AF, AT, EF and ET in low selenium group when compared with control group. CONCLUSIONS: Low selenium undermines the ultrastructure of chondrocytes, inhibits the normal development of cartilage and the expression of CS sulfated enzymes.

Impact of Interfacial Composition on Lipid and Protein Co-Oxidation in Oil-in-Water Emulsions Containing Mixed Emulisifers.

The impact of interfacial composition on lipid and protein co-oxidation in oil-in-water emulsions containing a mixture of proteins and surfactants was investigated. The emulsions consisted of 5% v/v walnut oil, 0.5% w/v whey protein isolate (WPI), and 0 to 0.4% w/v Tween 20 (pH 3 and pH 7). The protein surface load, magnitude of the ξ-potential, and mean particle diameter of the emulsions decreased as the Tween 20 concentration was increased, indicating the whey proteins were displaced by this nonionic surfactant. The whey proteins were displaced from the lipid droplet surfaces more readily at pH 3 than at pH 7, which may have been due to differences in the conformation or interactions of the proteins at the droplet surfaces at different pH values. Emulsions stabilized by whey proteins alone had relatively low lipid oxidation rates when incubated in the dark at 45 °C for up to 8 days, as determined by measuring lipid hydroperoxides and 2-thiobarbituric acid-reactive substances (TBARS). Conversely, the whey proteins themselves were rapidly oxidized, as shown by carbonyl formation, intrinsic fluorescence, sulfhydryl group loss, and electrophoresis measurements. Displacement of whey proteins from the interface by Tween 20 reduced protein oxidation but promoted lipid oxidation. These results indicated that the adsorbed proteins were more prone to oxidation than the nonadsorbed proteins, and therefore, they could act as better antioxidants. Protein oxidation was faster, while lipid oxidation was slower at pH 3 than at pH 7, which was attributed to a higher antioxidant activity of whey proteins under acidic conditions. These results highlight the importance of interfacial composition and solution pH on the oxidative stability of emulsions containing mixed emulsifiers.

Ultrasound improving the physical stability of oil-in-water emulsions stabilized by almond proteins.

BACKGROUND: Vegetable proteins are increasingly used to stabilize oil-in-water (O/W) emulsions. However, emulsions are thermodynamically unstable. Recently, high-intensity ultrasound (US) has been used to enhance the stability of emulsions. For these reasons, and considering almond (Prunus dulcis L.) as a good source of high-quality proteins, the aim of this work was to investigate the effect of US treatment on the stability of pre-emulsification O/W emulsions coated with almond protein isolate (API). RESULTS: The influence of API concentration (0.25-2.0 g L-1 ), ion strength (0-500 mmol L-1 NaCl), and pH (3.0-7.0) on the stability of US-treated emulsions was evaluated. US treatment (200-600 W, 25 kHz, 15 min) led to a significant reduction in the particles size of droplets in emulsions, increased critical osmotic pressure and additional protein interfacial adsorption, and thus the formation of more stable emulsions. The more unfolded and random coil structures of the proteins were detected at higher US power, facilitating protein interfacial adsorption. Increasing API concentrations resulted in higher stability of US-treated emulsions against untreated counterparts. The US-treated emulsions were more resistant to salt than untreated samples. In the range from pH 3.0 to7.0, US treatment also enhanced the physical stability of emulsions compared with untreated emulsions. CONCLUSION: US technology could be applied to produce more stable O/W food emulsions stabilized by proteins. © 2018 Society of Chemical Industry.

The prooxidant activity of salts on the lipid oxidation of lecithin-stabilized oil-in-water emulsions.

Salts reduction/substitution have gained a lot interest from food industry since the US Food and Drug Administration (FDA) has issued a draft guidance for salt reduction. However how changes of salts in food formulation could influence lipid oxidation is still not fully understood. Using oil-in-water emulsions stabilized by a natural emulsifier - lecithin at pH 7.0 as a model system, this study evaluated how salts affect the physical parameters of the emulsion, the chelating activity of lecithin and thus the lipid oxidation of these emulsions. Results showed that salts increased the particle size, the negative charge of the oil droplets, and the amount of iron chelated by lecithin. Lipid oxidation lag phases were shortened by addition of salts, by 1 day and 2 days for lipid hydroperoxides and thiobarbituric acid reactive substances measurements respectively. These results provide some new insights on the mechanisms of how salts could affect the lipid oxidation of food emulsions.

Long-Term Selenium-Deficient Diet Induces Liver Damage by Altering Hepatocyte Ultrastructure and MMP1/3 and TIMP1/3 Expression in Growing Rats.

The effects of selenium (Se)-deficient diet on the liver were evaluated by using growing rats which were fed with normal and Se-deficient diets, respectively, for 109 days. The results showed that rats fed with Se-deficient diet led to a decrease in Se concentration in the liver, particularly among male rats from the low-Se group. This causes alterations to the ultrastructure of hepatocytes with condensed chromatin and swelling mitochondria observed after low Se intake. Meanwhile, pathological changes and increased fibrosis in hepatic periportal were detected by hematoxylin and eosin and Masson's trichrome staining in low-Se group. Furthermore, through immunohistochemistry (IHC) staining, higher expressions of metalloproteinases (MMP1/3) and their tissue inhibitors of metalloproteinases (TIMP1/3) were observed in the hepatic periportal of rats from the low-Se group. However, higher expressions of MMP1/3 and lower expressions of TIMP1/3 were detected in hepatic central vein and hepatic sinusoid. In addition, upregulated expressions of MMP1/3 and downregulated expressions of TIMP1/3 at the messenger RNA (mRNA) and protein levels also appeared to be relevant to low Se intake. In conclusion, Se-deficient diet could cause low Se concentration in the liver, alterations of hepatocyte ultrastructure, differential expressions of MMP1/3 and TIMP1/3 as well as fibrosis in the liver hepatic periportal.

[Effect of low-selenium diet on expressions of CCR7, CD206 and CD163 in the liver and kidney of rats].

OBJECTIVE: To investigate the effect of low-selenium diet on the liver and kidneys of rats and explore the role of macrophage polarization into M1 and M2 phenotypes in liver and kidney injuries. METHODS: Twenty-four rats (12 female and 12 male) were randomly divided into control group and low-selenium group and fed with normal chow (dietary selenium of 0.18 mg/kg) and low-selenium diet (dietary selenium of 0.02 mg/kg) for 109 days. After the feeding, the rats were sacrificed for HE staining to observe liver and kidney pathologies, and immunohistochemistry was performed for analyzing CCR7, CD206, CD163-positive cell numbers in the liver and kidneys. RESULTS: The rats in low-selenium group showed severer fibrosis in the liver and kidney than the control group. In either male or female rats in low-selenium group, CCR7 and CD206 expressions in the liver were comparable with those in control group, but CD163 expression was lower than that in the control group (P<0.05 for both female and male rats). In the kidney, the proximal tubule showed a slightly higher while the distal tubule showed a slightly lower CCR7 expression in low selenium group than in the control group (P>0.05). In low-selenium group, a significantly lower CD163 expression in the distal tubule and a significantly higher CD206 expression in the proximal tubule were noted as compared with the control group (P<0.05 in both female and male rats). Compared with the control rats, the male rats in low-selenium group, but not the female rats, showed a significantly lower CD163 expression in the proximal tubule of the kidney (P<0.05); the female but not the male rats in low-selenium group show a higher CD206 expression in the distal tubule (P<0.05). CONCLUSION: Low-selenium diet can cause liver and kidney fibrosis in rats and may inhibit macrophage activation into the M2 phenotype.

Selenium deficiency induced damages and altered expressions of metalloproteinases and their inhibitors (MMP1/3, TIMP1/3) in the kidneys of growing rats.

Selenium is an essential trace element for the maintenance of structures and functions of kidney. To evaluate the effects of low selenium on the kidneys of growing rats, newborn rats were fed with selenium deficient and normal diets respectively for 109 days. As a result, rats fed with low selenium diets resulted in a decline in the body weight and the concentration of selenium in the kidney, especially the male rats from the low selenium groups. Moreover, the ultrastructure of glomerulus and tubules were damaged in low selenium group: the glomeruli were observed with hyperplasia of mesangial cells, fusion of podocyte foot processes and thickening of basement membrane; and the tubules were observed with vacuolar degenerated epithelial cells, increased edema fluid or protein solution between cells, microvilli edema, increased cell gaps and decreased cell links. Furthermore, the pathological changes in selenium deficient group included the increase of fibers around renal hilum aorta and in the renal collecting duct, and shed of cells in the proximal convoluted tubules. In addition, up-regulated expressions of matrix metalloproteinases (MMP1/3) and down-regulated expressions of their inhibitors (TIMP1/3) at the mRNA and protein levels were also appeared to be relevant to low selenium. The results suggested that low selenium in diet may cause low selenium concentration in the kidney of growing rat and lead to damages of the ultrastructure and extracellular matrix (ECM) of kidney.

Influence of aqueous phase emulsifiers on lipid oxidation in water-in-walnut oil emulsions.

Effects of selected aqueous phase emulsifiers on lipid oxidative stability of water-in-walnut oil (W/O) emulsions stabilized by polyglycerol polyricinoleate (PGPR) were evaluated. The formation of primary oxidation products (lipid hydroperoxides) and secondary oxidation products (headspace hexanal) increased with increasing dodecyltrimethylammonium bromide (DTAB) concentration (0.1-0.2 wt % of emulsions). In contrast, the addition of sodium dodecyl sulfate (SDS) in the aqueous phase reduced lipid hydroperoxide and hexanal formation. In addition, the presence of Tween 20 in the aqueous phase did not significantly influence lipid oxidation rates in W/O emulsions compared to the control (without Tween 20). Whey protein isolate (WPI) was observed to inhibit lipid oxidation in the W/O emulsions (0.05-0.2 wt % of emulsions). Aqueous phase pH had an important impact on the antioxidant capability of WPI, with higher pH improving its ability to inhibit lipid oxidation. The combination of WPI and DTAB in the aqueous phase suppressed the prooxidant effect of DTAB. The combination of WPI and SDS resulted in improved antioxidant activity, with inhibition being greater at pH 7.0 than at pH 3.0. These results suggest that the oxidative stability of W/O emulsions could be improved by the use of suitable emulsifiers in the aqueous phase.

[Therapeutic effect of capsule xinganbao on schistosomal hepatocirrhosis].

A total of 118 patients with schistosomal hepatocirrhosis were divided into an experiment group (62 cases) and a control group (56 cases), and the former received capsule Xinganbao and the latter received conventional medicine for 6 months. The reductions of serum HA, LN and PCIII were more notable in the experiment group than in the control group (P < 0.05). The clinical symptoms, physical signs, and liver function improved in both groups, and there were no significant differences between two groups. In conclusions, capsule Xinganbao has a good effect on hepatic fibrosis.