Intra-Amniotic Administration (Gallus gallus) of Cicer arietinum and Lens culinaris Prebiotics Extracts and Duck Egg White Peptides Affects Calcium Status and Intestinal Functionality

Abstract

Calcium (Ca) is one of the most abundant inorganic elements in the human body and has many important physiological roles. Prebiotics and bioactive peptides are two important substances used to promote calcium uptake. However, the difference in mechanisms of the calcium uptake from these two supplements is not clear. By using the Gallus gallus model and the intra-amniotic administration procedure, the aim of this study was to investigate whether Ca status, intestinal functionality, and health-promoting bacterial populations were affected by prebiotics extracted from chickpea and lentil, and duck egg white peptides (DPs). Eleven groups (non-injected; 18 MΩ H2O; 4 mmol/L CaCl2; 50 mg/mL chickpea + 4 mmol/L CaCl2; 50 mg/mL lentil + 4 mmol/L CaCl2; 40 mg/mL DPs + 4 mmol/L CaCl2; 5 mg/mL Val-Ser-Glu-Glu (VSEE) + 4 mmol/L CaCl2; 50 mg/mL chickpea; 50 mg/mL lentil; 40 mg/mL DPs; 5 mg/mL VSEE) were utilized. Upon hatch, blood, cecum, small intestine, liver and bone were collected for assessment of serum bone alkaline phosphate level (BALP), the relative abundance of intestinal microflora, expression of Ca-related genes, brush border membrane (BBM) functional genes, and liver and bone mineral levels, respectively. The BALP level increased in the presence of lentil, DPs and VSEE (p < 0.05). The relative abundance of probiotics increased significantly (p < 0.05) by VSEE + Ca and chickpea. The expression of CalbindinD9k (Ca transporter) increased (p < 0.05) in Ca, chickpea + Ca and lentil + Ca groups. In addition, the brush border membrane functionality genes expressions increased (p < 0.05) by the chickpea or lentil extracts. Prebiotics and DPs beneficially affected the intestinal microflora and duodenal villus surface area. This research expands the understanding of the prebiotics’ properties of chickpea and lentil extracts, and peptides’ effects on calcium metabolism and gut health.

The In Ovo Feeding Administration (Gallus Gallus)—An Emerging In Vivo Approach to Assess Bioactive Compounds with Potential Nutritional Benefits

“…Recently, the intra-amniotic administration model was used to evaluate the effects of prebiotics and duck egg white peptides on the promotion of calcium uptake [ 42 ]. It was found that the prebiotics and peptides increased the relative abundance of beneficial probiotics, the intestinal villus surface area, and goblet cell diameters, as well as regulated the calcium-related gene expressions.…”

“…Prebiotics from chickpea and lentil improve calcium bioavailability by promotion of gut beneficial prebiotics levels, the enlargement of gut villus surface area, and the improvement of BBM functionality. Duck egg white peptides promote calcium uptake through the reaction with calcium to act as calcium carriers and maintain gut health [ 42 ].…”

“…In this context, the in ovo feeding of DiNovo (extract of Laminaria species of seaweed) significantly increased the width of duodenal villi and the depth of the crypts [ 37 ]. The villi surface area was also observed to increase post intra-amniotic administration of raffinose and stachyose [ 41 , 46 ], chickpea and lentil prebiotics [ 42 ], egg white peptides [ 42 ], symbiotic (inulin, Enterococcus faecium ) [ 48 ], mannan oligosaccharides [ 56 ], carbohydrates (dextrin, maltose, sucrose) [ 57 ], and zinc-methionine [ 58 ]. However, in ovo injected probiotics (inulin, transgalactooligosaccharides, Lactococcus lactis ) did not affect the villi heights, but rather changed the crypts’ depth [ 49 ].…”

Nicotianamine-chelated iron positively affects iron status, intestinal morphology and microbial populations in vivo (Gallus gallus)

“…serum, and all animal tissues was determined by nitric/perchloric acid digestion as previously described 38 followed by inductively coupled plasma-optical emission spectrometry (ICP-OES) using a Thermo iCAP 6500 series (Thermo Jarrell Ash Corp., Franklin, MA, USA). Fe-NA solution (Fe-NA); (6) 'Control' white flour extract (C WF); (7) 'Biofortified' white flour extract (B WF) and eggs were subsequently incubated for four days until hatch as described in 79,80 . Chicks were euthanized by CO 2 exposure after hatching and all tissues were collected.…”

“…Lactobacillus, Bifidobacterium, Escherichia, and Clostridium density in intraamniotic administration treatment groups was determined as previously described 79 . In brief, cecal contents were homogenized with phosphate-buffered saline (PBS, 9 ml), centrifuged and the pellet resuspended in ethylenediaminetetraacetic acid (EDTA, 50 mM) and treated (37 °C, 45 min) with lysozyme (10 mg/mL, Sigma Aldrich CO., St. Louis, MO, USA).…”