Synbiotics are synergistic combinations of prebiotics and probiotics. In chickens, synbiotics can be delivered in ovo to expedite colonization of the gut by beneficial bacteria. We therefore aimed to design synbiotics in vitro and validate them in broiler chickens upon in ovo delivery. The probiotic components of the synbiotics were Lactobacillus salivarius and Lactobacillus plantarum. Their growth was assessed in MRS medium supplemented with different prebiotics. Based on in vitro results (hatchability and growth curve), two synbiotics were designed: S1 –Lactobacillus salivarius with galactooligosaccarides (GOS) and S2 –Lactobacillus plantarum with raffinose family oligosaccharides (RFO). These synbiotics were delivered to Cobb broiler chicken embryos on day 12 of incubation at optimized doses (105 cfu egg-1 of probiotic, 2 mg egg-1 of prebiotic). Post hatching, 2,400 roosters were reared (600 individuals group-1 divided into eight replicate pens). Microbial communities were analyzed in ileal and cecal digesta on day 21 using FISH. Gene expression analysis (IL1β, IL4, IL6, IL8, IL12, IL18, IFNβ, and IFNγ) was performed on days 7, 14, 21, and 42 for the spleen and cecal tonsils with RT-qPCR. Body weight and feed intake of the roosters did not differ by the treatments. Microbial populations of Lactobacillus spp. and Enterococcus spp. in the ileum were higher in S1 and S2 than in the control. In the cecum, the control had the highest bacterial counts. S1 caused significant up-regulation of IL6, IL18, IL1β, IFNγ, and IFNβ in the spleen on day 21 and IL1β on day 7 (P < 0.05). In cecal tonsils, S1 caused significant down-regulation of IL12, IL8, and IL1β on day 42 and IFNβ on day 14 (P < 0.05). S2 did not elicit such patterns in any tissues investigated. Thus, we demonstrate that divergent effects of synbiotics in broiler chickens were reflected in in vitro tests.
A simple method for the isolation and purification of alpha-galactosides, raffinose family oligosaccharides (RFOs), from legumes has been developed. The method includes (i) imbibition of seeds, (ii) extraction with 50% ethanol, (iii) precipitation of RFOs, (iv) purification of RFOs on diatomaceous earth and charcoal, and (v) cation-exchange chromatography. The described method allows one to obtain high purity RFO preparations (90% for lentil and 80% for pea seeds, determined by HPLC-RI analysis) in the form of white, fine powder. Yields of alpha-galactosides isolated from 100 g of seeds of lentil and pea were 5.6 and 4.3 g, respectively.
Three cultivars of broccoli seeds (Brassica oleracea var. italica), cv. Tiburon, cv. Belstar and cv. Lucky, and two cultivars of radish seeds (Raphanus sativus), cv. Rebel and cv. Bolide, were germinated for three and five days and safety aspects such as microbiological counts and biogenic amines were investigated. Cytotoxicity evaluation was also carried out. Broccoli and radish sprouts contained numbers of mesophilic, psychrotrophic, total and faecal coliform bacteria which are the usual counts for minimally processed germinated seeds. Putrescine, cadaverine, histamine, tyramine, spermidine and spermine increased during sprout production although these levels were below those permitted by legislation (5 mg/100 g of edible food). Broccoli and radish sprouts demonstrated no toxic effects on proliferation and viability of HL-60 cells and should be included in our diets as healthy and safe fresh foods.
The objective of our studies were seeds of two lupin species Lupinus luteus L. and Lupinus angustifolius L. cvs. Lord and Graf respectively. Lupin seeds were germinated at 15 and 24°C and during two, three and four days. In the lupin sprouts antinutritional factors: alkaloids and raffinose family oligosaccharides (RFOs) and five nitrogen fractions: non protein (Nnp), albumin (A), globulin (G), glutelin and prolamin (Gt+P) and nitrogen residue fraction (Nr) were determined. The level of these compounds was compared with the proper ones of initial material (not germinated seeds). These studies showed that the germination process clearly affects the decrease of antinutritional factors: RFOs and alkaloids. The decrease level of these compounds depended on such factors like, lupin species and used germination conditions. It was found on the base of nitrogen analysis of particular protein fractions that the germination process of lupin seeds causes deep quantitative and qualitative changes in fractional composition of lupin proteins. It especially concerns the decrease of globulin and residual fraction content and distinct increase of Nnp fraction. The changes in other fractions were not so unequivocal in comparison with the mentioned above and depended on lupin species, temperature and time of germination. Qualitative changes of A, G and Gt+P fractions caused by germination were confirmed by gel electrophoresis (SDS-PAGE). The amino acid analysis of seeds and sprouts of Nnp fractions showed an increased content of Asp, Ser, Ala, Pro non essential amino acids (NEAA), and Val, Met, iLeu, Leu, Thr essential amino acids (EAA). Simultaneously a decrease of Glu, Arg (NEAA), Phe, Lis, Cys (EAA) contents was observed. Generally the germination process causes the decrease of total NEAA and an increase of total EAA in Nnp fractions of both lupin species.
The growth and physiological responses of the rats to diet supplemented with raw and -fermented yellow and blue lupin seeds were determined. The diets containing soya bean meal, raw and fermented blue and yellow lupin were administered to eight rats in each diet group for four weeks. Yellow lupin seeds in the diets of rats improved significantly (p<0.05) feed intake, protein digestibility, body mass gain and protein efficiency ratio in comparison with blue lupin seeds. On the contrary, blue lupin seeds affected significantly (p<0.05) gastrointestinal fermentation processes in comparison with yellow lupin seeds. Fermentation of lupin seeds increased crude protein content and reduced phytate and oligosaccharide content. In the fermented products, a higher number of lactic acid bacteria and yeasts but reduced number of coliform bacteria was found. Fermentation by positively (p<0.05) affected protein digestibility of feed, body mass gain and protein efficiency ratio of rats, as well as the activity of some bacterial enzymes and cholesterol concentrations in the blood serum.
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