Intra-Amniotic Administration—An Emerging Method to Investigate Necrotizing Enterocolitis, In Vivo (Gallus gallus)

Abstract

Necrotizing enterocolitis (NEC) is a severe gastrointestinal disease in premature infants and a leading cause of death in neonates (1–7% in the US). NEC is caused by opportunistic bacteria, which cause gut dysbiosis and inflammation and ultimately result in intestinal necrosis. Previous studies have utilized the rodent and pig models to mimic NEC, whereas the current study uses the in vivo (Gallus gallus) intra-amniotic administration approach to investigate NEC. On incubation day 17, broiler chicken (Gallus gallus) viable embryos were injected intra-amniotically with 1 mL dextran sodium sulfate (DSS) in H2O. Four treatment groups (0.1%, 0.25%, 0.5%, and 0.75% DSS) and two controls (H2O/non-injected controls) were administered. We observed a significant increase in intestinal permeability and negative intestinal morphological changes, specifically, decreased villus surface area and goblet cell diameter in the 0.50% and 0.75% DSS groups. Furthermore, there was a significant increase in pathogenic bacterial (E. coli spp. and Klebsiella spp.) abundances in the 0.75% DSS group compared to the control groups, demonstrating cecal microbiota dysbiosis. These results demonstrate significant physiopathology of NEC and negative bacterial–host interactions within a premature gastrointestinal system. Our present study demonstrates a novel model of NEC through intra-amniotic administration to study the effects of NEC on intestinal functionality, morphology, and gut microbiota in vivo.

Constitutive overexpression of a nicotianamine synthase gene in bread wheat and in vivo assessment of iron and zinc bioavailability

“…LC-MS/MS analysis of LA, DGLA, and d6DGLA was performed using a Luna C18 S1. RT-qPCR reactions were performed using the Bio-RadCFX96 Touch (Hercules, CA, USA), as previously described 24,51,52,82 . All reactions were performed in duplicate under the following optimal conditions: initial denaturation at 95˚C for 30 s, 40 cycles of denaturation at 95˚C for 15 s, various annealing temperatures (Supplementary Table S1) for 30 s and elongation at 60˚C for 30 s. Gene expression levels were obtained from Ct values based on the "second derivative maximum" as computed by the Bio-Rad CFX Maestro Software (Bio-Rad, Hercules, CA, USA) and normalized to 18S expression.…”

“…The G. gallus system is sensitive to minor modulations in dietary mineral concentration/content and is therefore frequently used to model human dietary mineral bioavailability and absorption 24,[48][49][50] . The intraamniotic administration method involves injecting extracts into the amniotic uid at day 17, which are entirely consumed by the embryo prior to hatch, and is widely used to assess the effect of bioforti ed foods on mineral transport and intestinal functionality and morphology [51][52][53][54] . Furthermore, humans and G. gallus share > 85% gene homology between intestinal brush border membrane proteins involved in mineral transport, further highlighting the relevance of the G. gallus in assessing mineral bioavailability 55 .…”

“…cDNA was stored at -20˚C until use. Primers used in the real-time polymerase chain reactions (RT-qPCR) were designed using Real-Time Primer Design Tool software (IDT DNA, Coralvilla, IA, USA) as was previously described24,51,52,82 . Primer sequences and accession numbers are shown in Supplementary Table…”

Emerging Dietary Bioactives in Health and Disease

“…researchers from multiple countries, including the USA, Brazil, United Kingdom, Italy, Norway, China, and Israel. The presented manuscripts cover a wide variety and range of topics in the field of dietary bioactives in health and disease, with emphasis on the investigation of the effects hydrolyzed protein of chia (Salvia hispanica L.) and Lacticaseibacillus paracasei on intestinal functionality, morphology, and bacterial populations, in vivo (Gallus gallus) [1]; the demonstration of the capacity of macauba (Acrocomia aculeata) pulp oil to prevent adipogenesis, inflammation and oxidative stress in mice fed a high-fat diet [2]; the demonstration of the capacity of kombuchas from green and black tea to modulate the gut microbiota and improve the intestinal health of wistar rats fed a high-fat highfructose diet [3]; the methodological preparation, characterization, wound healing, and cytotoxicity assay of pegylated nanophytosomes loaded with 6-gingerol [4]; the discussion of empire apple (Malus domestica) juice, pomace, and pulp and the modulation of intestinal functionality, morphology, and bacterial populations in vivo (Gallus gallus) [5]; the effect of chia (Salvia hispanica L.) associated with high-fat diet on the intestinal health of wistar rats [6]; the demonstration of the capacity of curcumin-added whey protein to positively modulate skeletal muscle inflammation and oxidative damage after exhaustive exercise [7]; the introduction of the novel intra-amniotic administration-an emerging method with which to investigate necrotizing enterocolitis in vivo (Gallus gallus) [8]; the investigation of the effect of black corn anthocyanin-rich extract (Zea mays L.) on cecal microbial populations in vivo (Gallus gallus) [9]; the demonstration of BRD9 inhibition by natural polyphenols which target DNA damage/repair and apoptosis in human colon cancer cells [10]; the alterations in intestinal brush border membrane functionality and bacterial populations following the intra-amniotic administration (Gallus gallus) of catechin and its derivatives [11]; the performance of a comparison of the effects of concord grape (Vitis labrusca L.) puree, juice, and pomace on intestinal morphology and functionality, and on bacterial populations in vivo (Gallus gallus) [12]; the demonstration of how the intra-amniotic administration (Gallus gallus) of genistein alters mineral transport, intestinal morphology, and gut microbiota [13]; the occurrence of the alterations in intestinal brush border membrane functionality and bacterial populations following intra-amniotic administration (Gallus gallus) of nicotinamide riboside and its derivatives [14]; a literature review and discussion of dietary trehalose as a bioactive nutrient [15]; and an analysis of how resistant maltodextrin consumption in a double-blind, randomized, crossover clinical trial induces specific changes in potentially beneficial gut bacteria [16]…”