Alginate Nanoencapsulated Synbiotic Composite of Pomegranate Peel Phytogenics and Multi-Probiotic Species as a Potential Feed Additive: Physicochemical, Antioxidant, and Antimicrobial Activities.

A synbiotic composed of alginate nanoencapsulated prebiotic (pomegranate peel phytogenics) and multi-species probiotics (Lactococcus lactis, Lactobacillus plantarum, Lactobacillus paracasei, and Saccharomyces cerevisiae) has been developed as a potential eco-friendly alternative to antibiotics. The physicochemical properties of the encapsulated synbiotic were evaluated, and its gastric and storage tolerance, as well as its antioxidant and antimicrobial activity, were tested and compared to that of the non-encapsulated synbiotic (free synbiotic). The results showed that the prebiotic pomegranate peel ethanolic extract contained seven phenolic compounds, with cinnamic being the most abundant (13.26 µL/mL). Sodium alginate-CaCl2 nanocapsules were effective in encapsulating 84.06 ± 1.5% of the prebiotic's phenolic compounds and 98.85 ± 0.57% of the probiotics. The particle size of the alginate-CaCl2 nanoencapsulated synbiotic was 544.5 nm, and the polydispersity index and zeta potential values were 0.593 and -12.3 mV, respectively. Thermogravimetric analysis showed that the alginate-CaCl2 nanoencapsulated synbiotic had high thermal stability at high temperatures, with only 2.31% of its weight being lost within the temperature range of 70-100 °C. The count of viable probiotics in the nanoencapsulated synbiotic was significantly higher than that in the free synbiotic after exposure to gastric acidity and storage for six months at room temperature. The percent inhibition values of the nanoencapsulated synbiotic and ascorbic acid (as a standard antioxidant) were comparable and significantly greater than those of the free synbiotic. The half-maximal inhibitory concentrations (IC50) of the nanoencapsulated synbiotic and ascorbic acid were significantly lower than those of the free synbiotic (3.96 ± 0.42 µg/mL and 4.08 ± 0.79 µg/mL for nanoencapsulated synbiotic and ascorbic acid, respectively, vs. 65.75 ± 2.14 µg/mL for free synbiotic). The nanoencapsulated synbiotic showed the highest significant antimicrobial activity against Escherichia coli (ATCC 8739). Both the nanoencapsulated and free synbiotics showed antimicrobial activity against Staphylococcus aureus (ATCC 6538), similar to that of gentamicin, although the nanoencapsulated synbiotic showed significantly higher inhibition activity compared to the free synbiotic. The nanoencapsulated synbiotic showed antimicrobial activity comparable to gentamicin against Pseudomonas aeruginosa (ATCC 90274), whereas the free synbiotic showed the least antimicrobial activity (p < 0.05). Both synbiotics showed significantly higher antimicrobial activity against Salmonella typhi (ATCC 6539) than gentamicin. Both synbiotics showed antifungal activity against Aspergillus niger and Aspergillus flavus, with a stronger effect observed for the nanoencapsulated synbiotic. However, the activity of both synbiotics was significantly lower than that of fluconazole (an antifungal drug).

Potential of montmorillonite modified by an organosulfur surfactant for reducing aflatoxin B1 toxicity and ruminal methanogenesis in vitro.

BACKGROUND: Montmorillonite clay modified by organosulfur surfactants possesses high cation exchange capacity (CEC) and adsorption capacity than their unmodified form (UM), therefore they may elevate the adverse impact of aflatoxin B1 (AFB1) on ruminal fermentation and methanogenesis. Chemical and mechanical modifications were used to innovate the organically modified nano montmorillonite (MNM). The UM was modified using sodium dodecyl sulfate (SDS) and grounded to obtain the nanoscale particle size form. The dose-response effects of the MNM supplementation to a basal diet contaminated or not with AFB1 (20 ppb) were evaluated in vitro using the gas production (GP) system. The following treatments were tested: control (basal diet without supplementations), UM diet [UM supplemented at 5000 mg /kg dry matter (DM)], and MNM diets at low (500 mg/ kg DM) and high doses (1000 mg/ kg DM). RESULTS: Results of the Fourier Transform Infra-Red Spectroscopy analysis showed shifts of bands of the OH-group occurred from lower frequencies to higher frequencies in MNM, also an extra band at the lower frequency range only appeared in MNM compared to UM. Increasing the dose of the MNM resulted in linear and quadratic decreasing effects (P < 0.05) on GP and pH values. Diets supplemented with the low dose of MNM either with or without AFB1 supplementation resulted in lower (P = 0.015) methane (CH4) production, ruminal pH (P = 0.002), and ammonia concentration (P = 0.002) compared to the control with AFB1. Neither the treatments nor the AFB1 addition affected the organic matter or natural detergent fiber degradability. Contamination of AFB1 reduced (P = 0.032) CH4 production, while increased (P < 0.05) the ruminal pH and ammonia concentrations. Quadratic increases (P = 0.012) in total short-chain fatty acids and propionate by MNM supplementations were observed. CONCLUSION: These results highlighted the positive effects of MNM on reducing the adverse effects of AFB1 contaminated diets with a recommended dose of 500 mg/ kg DM under the conditions of this study.

Effect of Nanoencapsulated Alginate-Synbiotic on Gut Microflora Balance, Immunity, and Growth Performance of Growing Rabbits.

A synbiotic comprising Saccharomyces cerevisiae yeast (SCY) and Moringa oleifera leaf extract (MOLE) has been encapsulated using nanotechnology. This duo is used as a dietary supplement for growing rabbits. Physicochemical analyses, in vitro antimicrobial activity, and gastrointestinal system evaluation were used to evaluate the quality of the nanofabricated synbiotic. The in vivo study was conducted using 40-day-old male growing rabbits (n = 16 rabbits/group) to evaluate the effect of the nanofabricated synbiotic on the health and growth performance of examined rabbits. Rabbits were equally allocated into four groups; (a) NCS, which received a basal diet supplemented with a noncapsulated 11 × 1012 CFU SCY + 0.15 g MOLE/kg diet, (b) LCS: those receiving a nanoencapsulated 5.5 × 1012 CFU SCY + 0.075 g MOLE/kg diet, (c) HCS: those receiving an 11 × 1012 CFU SCY + 0.15 g MOLE/kg diet, and (d) CON: those receiving a basal diet without treatment (control). The treatments continued from day 40 to day 89 of age. During the experimental period, growth performance variables, including body weight (BW), feed consumption, BW gain, and feed conversion ratio were recorded weekly. Blood samples were collected on day 40 of age and immediately before the start of the treatments to confirm the homogeneity of rabbits among groups. On day 89 of age, blood samples, intestinal, and cecal samples were individually collected from eight randomly selected rabbits. The size and polydispersity index of the nanofabricated synbiotic were 51.38 nm and 0.177, respectively. Results revealed that the encapsulation process significantly improved yeast survival through the gastrointestinal tract, specifically in stomach acidic conditions, and significantly increased in vitro inhibitory activities against tested pathogens. Furthermore, treatments had no negative effects on hematobiochemical variables but significantly improved levels of blood plasma, total protein, and insulin-like growth factor-l. Compared to the CON, NCS, and LCS treatments, the HCS treatment increased the amount of intestinal and cecal yeast cells (p < 0.05) and Lactobacillus bacteria (p < 0.05) and decreased number of Salmonella (p < 0.05) and Coliform (p = 0.08) bacteria. Likewise, both LCS and HCS significantly improved the small intestine and cecum lengths compared to CON and NCS. The HCS treatment also significantly improved BW gain and feed conversion compared to CON treatment, whereas the NCS and LCS treatments showed intermediate values. Conclusively, the nanoencapsulation process improved the biological efficiency of the innovative synbiotic used in this study. A high dose of encapsulated synbiotic balanced the gut microflora, resulting in the growth of rabbits during the fattening period.

Effects of Organic Selenium on the Physiological Response, Blood Metabolites, Redox Status, Semen Quality, and Fertility of Rabbit Bucks Kept Under Natural Heat Stress Conditions.

Heat stress can impair the general health of rabbit bucks by disturbing physiological homeostasis with negative consequences in animal welfare and remarkable decline in reproductive performance. Selenium (Se) can control a number of vital biological processes. Thus, the effects of organic selenium (OSe) supplementation on the blood metabolites, redox status, semen quality, testicular histology, seminal plasma protein profile, and fertility of rabbit bucks kept under natural heat stress conditions were studied. Adult V-line male rabbits were fed a basal diet supplemented with 0.3 mg OSe/kg dry matter (DM) diet (OSe, n = 9) or not (control, CON, n = 9) for 12 weeks. The results showed that rabbits fed the OSe diet had 73.68 and 68.75% higher (P < 0.05) OSe concentrations in the blood serum and seminal plasma, respectively, than rabbits fed the CON diet. The OSe diet significantly decreased the rectal temperature and respiration rate and significantly increased the blood serum concentrations of total protein, albumin, glucose, and glutathione peroxidase compared to the CON diet. Rabbits fed the OSe diet had lower reaction times (12.53 vs. 5.84 s, ± 0.79, P < 0.01) and higher total functional sperm counts (116.74 vs. 335.23 × 106/ml, ± 24.68, P < 0.001) and percentages of integrated sperm membranes (60.38 vs. 79.19%, ± 1.69, P < 0.01) than rabbits fed the CON diet. Rabbits fed the OSe diet had higher (P < 0.01) contents of seminal plasma total protein, albumin, alanine transaminase, fructose, and total antioxidant capacity and lower (P < 0.001) malondialdehyde (MDA) levels than those fed the CON diet. Rabbits fed the OSe diet had sperm cells with higher levels of integrated DNA than those fed the CON diet. The seminal plasma of rabbits fed the OSe diet contained four new proteins, with molecular weights of 19.0, 21.5, 30.0, and 44.0 kDa. The kindling rates, litter size, and weight at birth of females mated with males fed the OSe diet were significantly higher than those of females mated with males fed the CON diet. In summary, the inclusion of 0.3 mg OSe/kg DM diet of naturally heat-stressed rabbit bucks countered the negative impacts of elevated environmental temperature on physiological homeostasis, semen quality, and fertility.