Search Results (12)

Exploring how early-life nutritional interventions may impact future generations, this study examines the inter- and transgenerational effects of in ovo injection of bioactive compounds on gene expression in the cecal tonsils and cecal mucosa using a chicken model. Synbiotic PoultryStar^® (Biomin) and choline were injected in ovo on the 12th day of egg incubation. Three experimental groups were established in the generation F1: (1) a control group (C) receiving 0.9% physiological saline (NaCl), (2) a synbiotic group (SYN) receiving 2 mg/embryo, and (3) a combined synbiotic and choline group (SYNCH) receiving 2 mg synbiotic and 0.25 mg choline per embryo. For the generations F2 and F3, the SYN and SYNCH groups were each divided into two subgroups: (A) those injected solely in F1 (SYNs and SYNCHs) and (B) those injected in each generation (SYNr and SYNCHr). At 21 weeks posthatching, cecal tonsil and cecal mucosa samples were collected from F1, F2, and F3 birds for transcriptomic analysis. Gene expression profiling revealed distinct intergenerational and transgenerational patterns in both tissues. In cecal tonsils, a significant transgenerational impact on gene expression was noted in the generation F3, following a drop in F2. In contrast, cecal mucosa showed more gene expression changes in F2, indicating intergenerational effects. While some effects carried into F3, they were less pronounced, except in the SYNs group, which experienced an increase compared to F2. The study highlights that transgenerational effects of epigenetic modifications are dynamic and unpredictable, with effects potentially re-emerging in later generations under certain conditions or fading or intensifying over time. This study provides valuable insights into how epigenetic nutritional stimulation during embryonic development may regulate processes in the cecal tonsils and cecal mucosa across multiple generations. Our findings provide evidence supporting the phenomenon of epigenetic dynamics in a chicken model. Full article

Background: In ovo stimulation introduces bioactive compounds, such as prebiotics, probiotics, or synbiotics into incubating eggs to enhance gut health and immune system development in chickens. This study aimed to determine the genetic and environmental effects modulating responses to in ovo stimulation in commercial broilers and Green-legged Partridge-like (GP) native chickens. Methods: Eggs were stimulated on day 12 of incubation with prebiotics (GOS—galactooligosaccharides), probiotics (Lactococcus lactis subsp. cremoris), or synbiotics (GOS + L. lactis), with controls being mock-injected. Hatched chicks were reared in group pens and challenged with lipopolysaccharide (LPS) on day 42 post-hatching. Cecal tonsils (CT) and spleens were harvested 2 h post-challenge. RT-qPCR was used to analyze the relative gene expression of cytokine genes: IL-1β, IL-2, IL-4, IL-6, IL-10, IL-12p40, and IL-17. Results: The results show that genotype influenced the expression of all immune-related genes, with broiler chickens exhibiting stronger innate inflammatory responses than native chickens. LPS induced both mucosal (CT) and systemic (spleen) immune responses in broilers but only systemic (spleen) responses in native chickens. Conclusions: In ovo stimulation had less of an impact on cytokine gene expression than LPS challenge. Broilers expressed higher inflammatory immune responses than GP native chickens. Full article

The present study aimed to test the synbiotic PoultryStar^® sol^US delivered in ovo to evaluate its effect on hatchability, productive performance and meat quality, compared to its post-hatch administration in water. On the twelfth day of embryonic incubation, 1200 fertile eggs were divided into synbiotic groups injected with 2 mg/embryo (T1) and 3 mg/embryo (T2), a saline group injected with physiological saline and an uninjected control group (C). After hatching, 120 male chicks/group were reared and chicks from the saline group were supplemented with the synbiotic via drinking water (T3). Hatchability was low in both T1 and T2 groups. Growth performance was not affected by the treatments. However, in the second rearing phase (15–36 days), birds from the C and T3 groups were heavier than T1 birds, due to a higher feed intake and daily weight gain. Neither route of synbiotic administration influenced final body weight (at 56 days), weight and yield of the carcass or commercial cuts. Physico-chemical properties, total lipid, cholesterol and fatty acid composition of breast muscle were not affected by the treatments. Considering its exploratory nature, this study has raised many questions that need further investigation, such as the bioactive combination and the effect on embryonic development. Full article

The aim of the study was to determine the effect of probiotics, prebiotics and synbiotics administered in ovo on selected morphological parameters of the small intestine (duodenum, jejunum, ileum) in broiler chickens (Ross 308) and native chickens (Green-legged Partridge, GP). On the 12th day of embryonic development (the incubation period), an aqueous solution of a suitable bioactive substance was supplied in ovo to the egg’s air cell: probiotic—Lactococcus lactis subsp. cremoris (PRO), prebiotic—GOS, galacto-oligosaccharides (PRE) or symbiotic—GOS + Lactococcus lactis subsp. cremoris (SYN). Sterile saline was injected into control (CON) eggs. After hatching, the chicks were placed in pens (8 birds/pen, 4 replicates/group) and housed for 42 days. On the last day of the experiment, all birds were individually weighed and slaughtered. Samples for histological analysis were taken directly after slaughter from three sections of the small intestine. In samples from the duodenum, jejunum and ileum, the height and width of the intestinal villi (VH) were measured and their area (VA) was calculated, the depth of the intestinal crypts (CD) was determined, the thickness of the muscularis was measured and the ratio of the villus height to the crypt depth (V/C) was calculated. On the basis of the obtained data, it can be concluded that the applied substances administered in ovo affect the production parameters and intestinal morphology in broiler chickens and GP. The experiment showed a beneficial effect of in ovo stimulation with a prebiotic on the final body weight of Ross 308 compared to CON, while the effect of the administered substances on the intestinal microstructure is not unequivocal. In GP, the best effect in terms of villi height and V/C ratio was found in the in ovo synbiotic group. Taking into account the obtained results, it can be concluded that chickens of different genotypes react differently to a given substance; therefore, the substances should be adapted to the genotype. Full article

The aim of the study was to analyse the effect of probiotics, prebiotics and synbiotics injected in ovo on day 12 of embryonic development on the microstructure of the superficial pectoral muscle (musculus pectoralis superficialis) from 42-day-old chickens of different genotypes: broilers (Ross 308) and general-purpose type (green-legged partridge (GP) chickens Zk-11, native chickens). Incubated eggs were divided into four groups (each genotype separately) depending on the substance injected in ovo: normal saline (C, control); Lactococcus lactis subsp. cremoris (PRO); galactooligosaccharides, GOS (PRE) or GOS + L. lactis (SYN). After hatching, chicks were placed in eight replicated pens (four pens/genotype group). There were eight birds per pen. In total, 64 birds were used in the experiment. Birds were slaughtered at the age of 42 days, and samples of superficial pectoral muscles were taken for analysis. The microstructure of the pectoral muscles was evaluated using the cryosectioning (frozen tissue sectioning) technique and staining with haematoxylin and eosin. Statistical analysis revealed that the in ovo injection of probiotics, prebiotics and synbiotics had no significant effect on the diameter of muscle fibres from chickens of the two genotypes. The number of fibres in the muscles from green-legged partridge chickens was about three-fold higher than the fibre density in the muscles from broiler chickens, with the fibre diameter being two-fold smaller. This fact may indicate a greater tenderness of meat from GP chickens compared to the meat from Ross 308 broilers. In the case of broilers, a prebiotic (GOS) was the most effective bioactive substance in reducing the number of histopathological changes. Considering muscles from GP chickens, the number of normal fibres was highest in birds treated with the probiotic. These findings indicate that the microstructural features of pectoral muscles depend not only on the type of the injected bioactive substance but also on the genotype of chickens. Full article

Probiotic supplements have been used to decrease the gut carriage of antimicrobial-resistant Enterobacterales through changes in the microbiota and metabolomes, nutrition competition, and the secretion of antimicrobial proteins. Many probiotics have shown Enterobacterales-inhibiting effects ex vivo and in vivo. In livestock, probiotics have been widely used to eradicate colon or environmental antimicrobial-resistant Enterobacterales colonization with promising efficacy for many years by oral supplementation, in ovo use, or as environmental disinfectants. In humans, probiotics have been used as oral supplements for infants to decease potential gut pathogenic Enterobacterales, and probiotic mixtures, especially, have exhibited positive results. In contrast to the beneficial effects in infants, for adults, probiotic supplements might decrease potentially pathogenic Enterobacterales, but they fail to completely eradicate them in the gut. However, there are several ways to improve the effects of probiotics, including the discovery of probiotics with gut-protection ability and antimicrobial effects, the modification of delivery methods, and the discovery of engineered probiotics. The search for multifunctional probiotics and synbiotics could render the eradication of “bad” Enterobacterales in the human gut via probiotic administration achievable in the future. Full article

Epigenetic regulation of gene expression is a form of interaction of the external environment on reading and transcription of genetic information encoded in nucleic acids. We provided evidence that early stimulation of the chicken microbiota with in ovo delivered synbiotics influenced gene expression and DNA methylation in the liver. Therefore, we hypothesize that the stimulation of microbiota by administering bioactive substances in ovo also affects the activity of miRNA in liver. For the analysis of miRNA activity, RNA was isolated from liver of adult broiler chicken and native chicken breed. The animals received a prebiotic, probiotic and synbiotic in ovo on day 12 of egg incubation. The analysis of miRNA expression was performed using the LNA method on a miRNA panel selected on the basis of previous microarray experiments. We have found increased miRNA expression activity after probiotic and synbiotic administration, especially in native chicken breed. Our results suggest that prebiotics reduce or do not affect miRNA activity. We have also shown that miRNA activity is regulated by the substance and genotype of the chicken. We can conclude that miRNAs constitute an important component of the molecular mechanism of host–probiotic interaction in liver. Full article

The effects of in ovo-delivered prebiotics and synbiotics on the lymphocyte subsets of the lymphoid organs in non-immunized 7-day-old broiler chickens and in non-immunized, sheep red blood cells (SRBC)-immunized, and dextran (DEX)-immunized 21- and 35-day-old birds were studied. The substances were injected on the 12th day of egg incubation: Prebiotic1 group (Pre1) with a solution of inulin, Prebiotic2 group (Pre2) with a solution of Bi²tos (non-digestive transgalacto-oligosaccharides), Synbiotic1 group (Syn1) with inulin and Lactococcus lactis subsp. lactis IBB SL1, and Synbiotic2 group (Syn2) with Bi²tos and Lactococcus lactis subsp. cremoris IBB SC1. In 7-day-old chicks, a decrease in T splenocytes was noticed in all groups. The most pronounced effect in 21- and 35-day-old birds was an increase in TCRγδ⁺ cells in Syn1 and Syn2 groups. A decrease in bursal B cells was observed in DEX-immunized Pre1 group (21-day-old birds), and in the Syn1 group in non-immunized and SRBC-immunized 35-day-old birds. An increase in double-positive lymphocytes was observed in Pre1 (35-day-old birds) and Pre2 (immunized 21-day-old birds) groups. In Pre1 and Syn1 groups (21- and 35-day-old), an increase in B splenocytes and a decrease in T splenocytes were observed. We concluded that Syn1 was the most effective in the stimulation of the chicken immune system. Full article

DNA methylation inhibits DNA transcription by the addition of methyl residues to cysteine within the CpG islands of gene promoters. The process of DNA methylation can be modulated by environmental factors such as intestinal microbiota. In poultry, the composition of the intestinal microbiota can be stimulated by in ovo delivery of synbiotics. The present study aims to determine the effect of Lactobacillus synbiotics delivered in ovo on the level of hepatic DNA methylation in broiler chickens. In ovo stimulation was performed on day 12 of egg incubation. Bioactive compounds delivered in ovo included (S1)—Lactobacillus salivarius with GOS and (S2)—Lactobacillus plantarum with RFO. Samples were collected from six individuals from each group on day 42 post-hatching. DNA methylation of five genes selected on the basis of the transcriptome data were analyzed using the qMSP method. Significant changes were observed in DNA methylation of genes in liver including ANGPTL4 and NR4A3, after S2 delivery. The obtained results confirm that the downregulation of metabolic gene expression in the liver mediated by in ovo stimulation had epigenetic characteristics. Full article

To better understand the effects of synbiotics administered at early stages of embryonic development in poultry, it is necessary to analyze direct effects (meat quality) and the molecular background. The molecular interpretation of poultry meat properties after in ovo administration of synbiotics remains to be reported. The purpose of the present study was to analyze the molecular background of meat quality based on gene expression and basic physiological parameters. Eggs were injected with (S1) Lactobacillus salivarius with galacto-oligosaccharides or (S2) Lactobacillus plantarum with raffinose family oligosaccharides. The pectoral muscle was collected at two time points (day 7 and day 42) and subjected to RNA isolation. Gene expression analysis was performed by RT-qPCR for a panel of eight genes associated with metabolism. The concentration of glucose and hormones (insulin, glucagon, and leptin (S1 p = 0.04)) was also increased. The obtained results showed that metabolic gene expression in the muscle was more differential due to synbiotic stimulation on day 7 (FST in S1 p = 0.03; PDK4 in S1 p = 0.02 and S2 p = 0.01; CEBPB in S1 p = 0.01 and S2 p = 0.008; PHKB in S1 p = 0.01; PRKAG3 in S1 p = 0.02) than on day 42 (PDK4 in S1 p = 0.04). On the basis of the results obtained, it can be concluded that in ovo stimulation with S1 triggered the most potent and favorable changes in the pectoral muscle gene expression in broiler chickens. Full article

The effect of the in ovo application of selected prebiotics and synbiotics on the humoral immune response against T-dependent (SRBC) and T-independent (dextran) antigens and delayed-type hypersensitivity (DTH) to phytohemagglutinin was studied. On the 12th day of incubation, 800 eggs (Ross 308) were divided into five groups and injected into the egg air chamber with prebiotic inulin (Pre1), Bi²tos (Pre2), a synbiotic composed of inulin and Lactococcus lactis subsp. lactis IBB SL1 (Syn1), a synbiotic composed of Bi²tos and L. lactis subsp. cremoris IBB SC1 (Syn2), and physiological saline (control group; C). The chickens were immunized twice at the 7th and 21st day of life with SRBC and dextran. A DTH test was performed on the 7th, 21st, and 35th day. The application of prebiotics and synbiotics had no significant effect on the humoral immune response. SRBC-immunized in ovo Pre1- and Pre2-treated chickens showed significantly higher serum IgG levels than the control. A significant effect on the DTH reaction was detected on the 7th (Pre1 < C) and 21st (Pre2 > Syn2) day. However; Bi²tos may transiently stimulate the cellular immune response on the 21st day. It may be concluded that the application of inulin in an egg air chamber on the 12th day of incubation may stimulate the secondary immune response. The inulin-treated group exhibited a lower mortality rate than the control group. Full article

The aim was to investigate the impact of an automatic in ovo injection of the raffinose family oligosaccharides (RFO) extracted from the seeds of Lupinus luteus L, on the chicken performance and resistance in a production environment. At day 12 of incubation, a total of 57,900 eggs (Ross 308) were divided into two groups: 1/ Control, injected with 0.9% NaCl and 2/ RFO group, injected with 1.9 mg/egg of the lupin seed extract, dissolved in 0.2 mL NaCl. The performance parameters, biochemical indices (lipid profile, hepatic parameters), gut histomorphology and duodenum structure, oxidative stability of the meat and microbiological counts of the major commensal microbiota species were analyzed. Mortality, body weight, and feed conversion ratio (FCR) were not affected. By day 42, several health indices were improved with RFO and were reflected in a beneficial lipid blood profile, increased villi surface and better combating opportunistic pathogens through reduction of Clostridia and decreased coccidia counts. The RFO increased meat oxidation, but only at the beginning of the storage. The RFO sourced from local legumes can be considered a promising prebiotic for broiler chickens. In ovo delivery of prebiotics and/or synbiotics should be further optimized as an important strategy for the earliest possible modulation of chicken resistance. Full article