The use of transgenic plants as delivery system for antigenic proteins is attractive for its simplicity and increases likelihood for local immune response at sites of infection. The aim of this study was to evaluate the protective effect of oral administration of tobacco seeds, expressing the FedA, the major protein of the F18 adhesive fimbriae, and B subunit of verocytotoxin, against verocytotoxin-producing E. coli (VTEC) strain in piglets. Forty-three early weaned piglets, were randomly divided into 4 experimental groups: 3 test groups and a control. Treatment groups orally received a bolus, with different dose of tobacco seeds on 0, 1, 2, 14 days post primary administration. After challenge, with 1*10(10) CFU of O138 Escherichia coli strain, piglets showed clinical scores significantly higher in the control group compared to orally immunized groups (P < 0.05) and the latter showed a faster recovery than in CG. In conclusion, oral administration of recombinant tobacco seeds expressing antigenic proteins against VTEC strains can induce a protective effect against challenger strain in piglets.

Quebracho (Qu) and chestnut (Ch) are natural sources of tannins and they are currently used in animal nutrition as feed ingredients. However, to date the bio-accessibility, antimicrobial, antioxidant, and intestinal epithelial cell stimulatory doses of Qu and Ch have not been determined. Our study investigates the antioxidant and E. coli F4+ and F18+ growth inhibitory activity of Qu, Ch, and their combinations after solubilization in water (to evaluate the already bio-accessible molecules) and after simulated gastro-intestinal digestion in vitro. The effect of an in vitro digested Ch and Qu combination was also tested on intestinal epithelial IPEC-J2 cells experimentally stressed with hydrogen peroxide (H2O2) and Dextran Sodium Sulfate (DSS). The results showed that undigested Qu and Ch alone, and in combination, exerted a valuable antioxidant capacity and E. coli F4+ and F18+ growth inhibitory activity. The concentration of 1200 µg/mL exhibited the highest E. coli growth inhibitory activity for all the samples tested. In addition, after in vitro digestion, Qu and Qu50%–Ch50% maintained E. coli growth inhibitory activity and a modest antioxidant capacity. Three hours pre-treatment with in vitro digested Qu50%–Ch50% counteracted the H2O2 and DSS experimentally-induced stress in the intestinal IPEC-J2 cells. Ch and Qu tannin extracts, particularly when combined, may exert E. coli F4+ and F18+ growth inhibitory activity and valuable antioxidant and cell viability modulation activities.

Gaucher disease, the most common genetic lysosomal disorder, is caused by the lack of functional acid beta-glucosidase (GCase) and is currently treated at a very high cost by enzyme replacement therapy. In an attempt to provide a safe and cost-effective production system, human placental GCase was produced and purified from transgenic tobacco seeds. Plant-derived recombinant GCase was found to be enzymatically active, uptaken by human fibroblasts and free of immunogenic xylose and fucose residues. This report demonstrates the potential of plant bioreactors in the large-scale production of injectable proteins required for lifelong therapy.

We evaluated the effects of leonardite supplementation, mainly composed of humic acids (HAs), as a functional feed additive in weaned piglets. One hundred and twenty piglets (Large Withe × Landrace) were weaned at 28 ± 2 days, and randomly divided into two groups (6 pens per group, 10 piglets per pen). After one week of adaptation, for 40 days groups were fed a control diet (CTRL) and an HA enriched diet (0.25% of leonardite; HAG). Body weight (BW), average daily feed intake (ADFI), average daily gain (ADG), feed conversion ratio (FCR) were measured throughout the experimental period. On the last day of the trial four piglets per pen were randomly selected and the blood was collected to evaluate the serum metabolic profile and diamine oxidase content.Chemical analyses showed that leonardite was characterized by a high content of ash 23.27% (as-fed basis), polyphenolic content of 35.18 ± 3.91 mg TAEq/g, and an antioxidant capacity of 73.31 ± 8.22 μmol TroloxEq/g.The HAG group showed an increase in BW, ADG and ADFI (P < 0.01) compared to the CTRL group during the experimental period. In terms of the serum metabolic profile, the HAG group showed a significant increase in total protein content (P < 0.001), albumin (P < 0.001), albumin/globulin ratio (P < 0.01), phosphatase alkaline (P < 0.01), calcium, phosphorus and magnesium (P < 0.05) compared to the CTRL group.A modulation in the serum lipid profile was recorded. The HAG group showed a decrease in total triglycerides (P < 0.05) with higher total cholesterol (P < 0.05), however only high-density lipoprotein showed a significant increase (P < 0.001) compared to the CTRL group. No significant differences in the amount of diamine oxidase were found between groups.In conclusion, leonardite inclusion in the diet at 0.25% was shown to have a positive effect on the serum lipid profile and animal growth. This thus suggests that leonardite can be considered as a new feed additive, which improves the health and performance of weaned piglets.

In pig livestock, alternatives to in-feed antibiotics are needed to control enteric infections. Plant extracts such as tannins can represent an alternative as a natural source of functional compounds. The aim of this study was to evaluate the in vitro digestibility and in vivo effects of oral supplementation of combined chestnut (Ch) and quebracho (Qu) tannins in order to establish if they can induce a positive effect on weaned piglets’ performance, metabolic status and fecal parameters. In vitro digestibility (dry matter, DM) of diets was calculated using a multi-step enzymatic technique. In vitro digested diet samples were further tested on an intestinal porcine enterocyte cell line (IPEC-J2). Weaned piglets (n = 120; 28 ± 2 day old) were randomly allotted to two groups (12 pens in total with 10 pigs per pen): control (Ctrl) and treatment (Ch/Qu). After one week of adaptation (day 0), 35-day-old piglets in the Ctrl group were fed a Ctrl diet and the Ch/Qu group were fed with 1.25% Ch/Qu for 40 days. Body weight and feed intake per pen were recorded weekly. At day 40, blood and fecal samples were collected. Principal metabolic parameters were evaluated from blood samples by enzymatic colorimetric analysis. Total phenolic compounds, urea, and ammonia in feces were analyzed (Megazyme International, Bray, Ireland). In vitro digestibility and cell viability assays showed that the inclusion of 1.25% Ch/Qu slightly reduced diet digestibility compared with the Ctrl diet, while intestinal cell viability was not altered with low concentrations of Ch/Qu digesta compared with Ctrl. In vivo results did not show any adverse effects of Ch/Qu on feed intake and growth performance, confirming that dietary inclusion of Ch/Qu at a concentration of 1.25% did not impair animal performance. The decreased diet DM digestibility in the Ch/Qu diet may cause increased serum concentration of albumin (Ctrl: 19.30 ± 0.88; Ch/Qu: 23.05 ± 0.88) and albumin/globulin ratio (Ctrl: 0.58 ± 0.04; Ch/Qu: 0.82 ± 0.04), but decreased creatinine (Ctrl: 78.92 ± 4.18; Ch/Qu: 54.82 ± 4.18) and urea (Ctrl: 2.18 ± 0.19; Ch/Qu: 0.95 ± 0.19) compared with Ctrl. Pigs in the Ch/Qu group contained higher (p < 0.05) concentrations of fecal phenolic compounds and nitrogen than the Ctrl group, while fecal ammonia and urea were not affected by tannins. In conclusion, Ch/Qu tannin supplementation did not influence growth performance. Although lower digestibility was observed in the diet supplemented with Ch/Qu tannins, Ch/Qu supplementation did not show any adverse effect on intestinal epithelial cell viability.