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Extensive mechanistic evidence to support the beneficial function of dietary phytobiotic applications for broiler performance, gut function and health is highly warranted. In particular, for isoquinoline alkaloids (IQ) the underlying mechanisms related to critical gut homeostasis components such as cytoprotection and gut barrier are scarce, especially for young broilers at the starter growth stage (d1-10). The aim of this study was to investigate the effect of a standardized blend of IQs on the relative gene expression of critical biomarkers relevant for antioxidant response and barrier function along the intestine of young broilers at the end of starter growth phase. For this purpose, 182 one-day-old Ross 308 broilers were allocated in 2 treatments with 7 replicates of 13 broilers each: control diet-no other additions (NC), and control diet containing a standardized blend of IQs at 200 mg/kg of diet (M) for the starter growth period (1-10d). The results revealed that the IQs blend significantly upregulated (P < 0.05) the expression of genes related to antioxidant response in all intestinal segments. Moreover, the IQs blend enhanced (P < 0.05) gut barrier components primarily at duodenal level. In conclusion, the blend of IQs beneficially affected critical pathway components relevant for the gut antioxidant capacity and barrier along the intestine of young broilers., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The effects of concurrent reduction of dietary metabolizable energy (ME) and crude protein (CP) levels combined or not with the dietary inclusion of a phytogenic feed additive (PFA) were studied using a nutrigenomics approach. In particular, the expression of 26 critical genes relevant for inflammation control (TLR pathway), cellular apoptosis (MAPK pathway) cell growth and nutrient metabolism (PI3K-Akt-mTOR pathway) was profiled along the broiler intestine. Two dietary types (L and H) differing in metabolizable energy and crude protein levels (L: 95% and H: 100% of optimal Cobb 500 recommendations for ME and CP requirements) supplemented or not with PFA (- or +) and their interactions (L-, L+, H-, H+) were evaluated. There were only 3 total interactions (mTOR, IL8, and HRAS P < 0.05) between diet type and PFA inclusion indicating limited concurrent effects. Diet type, L upregulated genes related with inflammation mainly in the jejunum, ileum, and cecum (P < 0.05) and MAPK pathway in the ileum and cecum (P < 0.05). Moreover, diet type L negatively affected the expression of genes related to PI3K-Akt-mTOR pathway mainly in duodenum and cecum (P < 0.05). On the other hand, PFA inclusion downregulated (P < 0.05) genes related with TLR signaling pathway (TLR2B, MyD88, TLR3, IL8, LITAF) along the intestine and MAPK pathway genes (APO1, FOS) in jejunum (P < 0.05). Finally, PFA supplementation regulated nutrient sensing and metabolism in the cecum in a manner perceived as beneficial for growth. In conclusion, the study results highlight that the reduced ME and CP specifications, especially in the absence of PFA, regulate inflammation, apoptosis and nutrient metabolism processes at homeostatic control levels that hinder maximizing the availability of dietary energy and nutrients for growth purposes. Inclusion of PFA helped to adjust the respective homeostatic responses and control to levels supporting broiler performance, especially at reduced specification diets., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

A 12-wk study was conducted to investigate the effects of a phytogenic premix (PP) inclusion level on production performance, and the expression of genes relevant for detoxification (aryl hydrocarbon receptor; AhR) and antioxidant capacity (Nuclear factor erythroid 2-related factor 2; Nrf2) in the ovaries of laying hens. The PP consisted of bioactive substances derived from ginger, lemon balm, oregano, and thyme substances (Anco FIT-Poultry). Depending on PP inclusion level (i.e., 0, 500, 750, 1,000, and 1,500 mg/kg diet) in the basal diet, 385 laying hens Hy-Line Brown, 20-wk-old were assigned into 5 treatments: CON, P500, P750, P1000, and P1500, with 7 replicates of 11 hens with ad libitum access to feed and water. Performance parameters were closely monitored on a weekly basis and analyzed in the following 3 experimental periods: 1 to 4 wk, 5 to 8 wk, and 9 to 12 wk of treatment administration (i.e., 21-24, 25-28, and 29-32 wk of layers age, respectively). At the end of the 8th and 12th wk of the experiment (i.e., 28 and 32 wk of layers age), a layer from each replicate was selected, euthanized, the ovaries sampled and stored deep frozen until gene expression analysis. Data were analyzed by ANOVA and means compared using Tukey's honest significant difference test. Polynomial contrasts tested the linear and quadratic effect of PP inclusion levels. Results revealed that PP inclusion, improved (P < 0.05) laying rate and egg mass, compared to CON. Increasing PP inclusion level enhanced laying rate and egg mass, linearly and quadratically and peaked at P1000 (P < 0.05). In the ovaries, the AhR pathway genes assessed were down-regulated (P < 0.05) mainly at P1000 and P750 treatments. In addition, PP related cytoprotective potential was demonstrated via beneficial changes seen for the majority of the Nrf2-pathway genes assessed with the P1000 displaying most significant differences from CON. Conclusively, new data highlighted beneficial cytoprotective effects of PP inclusion on layer ovaries and documented further layer performance, with the inclusion level of 1000 mg PP/kg diet being the most prominent., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

This study evaluated the effect of reduced dietary energy (ME) and crude protein (CP) levels along with inclusion of a phytogenic feed additive (PFA) on gut microbiota composition and gene expression of Toll-like receptor(s) (TLR), tight junction proteins, and inflammatory cytokines expressed in secondary lymphoid organs. Depending on dietary ME and CP level down regulation and the inclusion or not of PFA at 125 mg/kg diet, 450 one-day-old male broilers were allocated in the following 6 treatments for 42 D according to a 3 × 2 factorial design: A: diet formulated optimally to meet broiler nutrient requirements; APh: A+PFA; B: suboptimal in ME and CP levels by 3%; BPh: B+PFA; C: suboptimal in ME and CP levels by 6%; CPh: C+PFA. Diet type and PFA supplementation were shown to affect mostly the mucosa-associated microbiota compared to the luminal ones. Ileal mucosa-associated total bacteria (PD= 0.005), Lactobacillus spp. (PD= 0.003), and Clostridium cluster XIVa (PD= 0.009) were affected by diet type with broilers fed diet B having lower levels compared to broilers fed diets A or C. Moreover, diet type affected cecal mucosa-associated Lactobacillus spp. (PD= 0.002) with broilers fed diet C having lower levels compared to broilers fed diets A or B. Supplementation with PFA resulted in higher levels of cecal mucosa-associated Bacteroides (PP= 0.031), Clostridium cluster IV (PP= 0.007), and Clostridium cluster XIVa (PP= 0.039). Diet type affected TLR2 (PD= 0.046) and claudin 5 (PD= 0.027) in cecal epithelium. Lower TLR2 (PP= 0.021) and higher zonula occludens 2 (PP= 0.031) relative gene expressions were seen in ileal epithelium following PFA supplementation. Moreover, in cecal epithelium, PFA supplementation resulted in lower TLR2 (PP < 0.001) and higher zonula occludens 2 (PP= 0.009), claudin 5 (PP= 0.005) and occludin (PP= 0.039) relative gene expressions. There were no significant diet type and PFA effects on cytokines in secondary lymphoid organs, except for a dietary effect on transforming growth factor beta 4 (PD= 0.023) in cecal tonsils. In conclusion, PFA inclusion beneficially modulated elements of gut microbiota, Toll-like signaling molecules and gut tight junction genes., (© 2018 Poultry Science Association Inc.)

The aim of this work was to assess the effect of dietary viable or heat inactivated probiotic forms (PF) combined or not with avilamycin (AV) used as a growth promoter, on broiler growth performance, nutrient digestibility, digestive enzyme activities, and expression of immune response related genes.Depending on the type of PF (i.e., no addition, viable, inactivated) and AV addition (no/yes), 450 one-day-old Cobb male broilers were allocated in the following 6 treatments according to a 3 × 2 factorial arrangement with 5 replicates of 15 broilers each for 6 wk: CoN: diet without any addition; CoN+A: combination of CoN with AV; ViP: viable PF - no AV; ViP+A: combination of ViP with AV; InP: inactivated PF - no AV; InP+A: combination of InP with AV.There were no interactions (P > 0.05) for overall performance parameters. In contrast, PF or AV addition improved BW gain (PPF= 0.015; PAV < 0.001), FCR (PPF < 0.001; PAV < 0.001) and production efficiency factor (PPF= 0.001; PAV= 0.001).Significant (PPF×AV ≤ 0.05) interaction effects regarding ileal digestibility (IAD) of DM and total tract apparent digestibility (TTAD) of DM and ether extracts (EE) were noted. In addition, PF affected IAD and TTAD of CP (PPF < 0.001, PPF= 0.004, respectively). Inactivated PF increased (PPR= 0.024) lipase activity in jejunal digesta.At spleen level InP and ViP+A down-regulated TGF-β4 (PPF × AV = 0.035) compared to CoN and ViP, whereas ViP+A up-regulated iNOS (PPF × AV = 0.022). An anti-inflammatory effect of live and inactive PF and/or AV addition at cecal tonsils was shown by iNOS down-regulation (PPF × AV= 0.015) compared to CoN. Furthermore, AV down-regulated IFN-γ (PAV= 0.002).In conclusion, viable probiotic, as well as inactivated probiotic alone or in combination with avilamycin, improved nutrient digestibility. All dietary additives affected growth performance positively and induced an anti-inflammatory response at cecal level., (© 2016 Poultry Science Association Inc.)

The dietary supplementation of Saccharomyces cerevisiae var. boulardii was evaluated in broilers challenged or not challenged with Salmonella Enteritidis (SE) using a 2 × 2 factorial arrangement. Depending on yeast inclusion at 0 (C) or 1 × 10⁹ cfu/kg diet (Y) and SE challenge (0 or log 6.3 cfu/bird) on d 15, the experiment had four treatments C, Y, C-SE, and Y-SE, respectively. Each treatment had seven replicate floor pens with 15 broilers. Growth performance responses were determined weekly and overall for the 5 week experimental period. Salmonella levels and prevalence in ceca, cloacae, and carcass skin were determined by culture procedures, while cecal microbiota was determined by real time PCR. Yeast supplementation had no effect (PY > 0.05) on growth performance. For the overall post SE-challenge period (i.e., wk 3 to wk 5), Salmonella reduced body weight gain (BWG) (PSE < 0.001), feed intake (FI) (PSE = 0.032), and the European production efficiency (EPEF) factor (PSE = 0.005). Broilers Y-SE had higher (P < 0.001) overall BW gain compared to C-SE ones. Overall mortality was 2.14% and did not differ (P > 0.05) between treatments. Reduced Salmonella levels in the cloacae (P = 0.014) and on the breast skin (P = 0.006) and lower prevalence on the neck skin (P = 0.007) were noted for treatment Y-SE compared to C-SE. Yeast supplementation did not have an effect (P > 0.05) on cecal microbiota composition at d 1 and d 21 post SE-challenge. On the contrary, SE-challenge reduced cecal levels of total bacteria (PSE = 0.002), E. coli (PSE = 0.006), Bifidobacterium spp. (PSE = 0.006), Bacteroides spp. (PSE = 0.010), and Clostridial populations belonging to cluster I and cluster XIVa, (PSE = 0.047 and PSE = 0.001, respectively) on d 1 post SE-challenge. At 21 d post SE-challenge, only the levels of cecal Lactobacillus spp. (PSE = 0.001) and Bifidobacterium spp. (PSE = 0.049) were reduced compared to the non SE-challenged groups. In conclusion, yeast supplementation in SE challenged broilers (Y-SE) was beneficial for growth performance and reduced Salmonella presence compared to C-SE ones. The disturbance of cecal microbiota balance by SE merits further investigation for potential implications in gut and overall bird health., (© 2015 Poultry Science Association Inc.)

The aim of this study was to investigate the effect of dietary probiotic inclusion level on mucin composition (monosaccharide ratio), mucosal morphometry, mucus layer staining intensity, and mucus layer thickness along the broiler intestinal tract. One-day-old male Cobb broilers were administered maize-soybean meal basal (BD) diets for 42 d and depending on the feed additive used, broilers were allocated into the following 5 experimental treatments: control C (BD, no additive), treatment P1 (10(8) colony forming units of probiotic/kg of BD), treatment P2 (10(9) cfu of probiotic/kg of BD), treatment P3 (10(10) cfu of probiotic/kg of BD), and treatment A (2.5 mg avilamycin/kg of BD). Intestinal samples from duodenum, ileum, and cecum of 14- and 42-d-old broilers were collected and analyzed. Mannose (Man) decreased linearly with increasing probiotic level in duodenum (P=0.015) and ileum (P=0.042) of 14-d-old broilers. N-Acetyl-glucosamine and galactose decreased linearly (P=0.012 and P=0.001, respectively), while fucose increased linearly (P<0.001) with increasing probiotic feed inclusion level in 42-d-old broiler cecum, with treatment A not differing from treatment C (P≥0.05). Cecal villus height and crypt depth increased linearly (P=0.016 and P=0.003, respectively) with probiotic inclusion level, with treatment A having higher (P≤0.05) values only from treatment C. Mucus layer thickness increased linearly with probiotic inclusion level in duodenum at 14 d and 42 d (P=0.007 and P=0.030, respectively). Finally, mucus layer staining intensity was influenced (P<0.001) by villus fragment (i.e., tip, midsection, and base) but not from the treatment, age, and intestinal segment examined. As a conclusion, this study provides evidence that probiotic inclusion level affects intestinal mucin monosaccharide composition, mucus layer thickness, and intestinal morphology in broilers.

The aim of this work was to investigate the effect of inclusion levels of a 5-bacterial species probiotic in broiler nutrition. Five hundred twenty-five 1-d-old male Cobb broilers were allocated in 5 experimental treatments for 6 wk. The experimental treatments received a corn-soybean coccidiostat-free basal diet and depending on the addition were labeled as follows: no addition (C), 10(8) cfu probiotic/kg of diet (P1), 10(9) cfu probiotic/kg of diet (P2), 10(10) cfu probiotic/kg of diet (P3), and 2.5 mg of avilamycin/kg of diet (A). Each treatment had 3 replicates of 35 broilers each. Treatment effects on broiler growth performance and biomarkers such as ileal and total tract nutrient digestibility, plasma Ig concentration, and cecal microflora composition were determined. Differences among treatments were considered significant when P < or = 0.05. Overall BW gain was significantly higher in treatment P1 (2,293 g) compared with P2 (2,163 g), C (2,165 g), and P3 (2,167 g), with A (2,230 g) being intermediate and not different from P1. Overall feed conversion ratio values were similar and significantly better for P1 (1.80) and A (1.80) compared with P2 (1.87), C (1.89), and P3 (1.92). Ileal apparent digestibility coefficients (ADC) of CP and ether extract were higher in A. Generally, treatments A and P1 showed an improved total tract ADC for DM, organic matter, ash, ether extract, and AME(n) values. The total tract ADC of CP was higher in P1, C, and P2. There were no differences between treatments regarding plasma Ig in 14- and 42-d-old broilers. Treatments P2 and P3 were effective at beneficially modulating cecal microflora composition. In particular, the lower cecal coliform concentration (log cfu/g of wet digesta) was seen in P2 (6.12) and P3 (4.90) in 14- and 42-d-old broilers, respectively, whereas at 42 d, P3 and P2 had the highest Bifidobacterium (8.31; 8.08) and Lactobacillus concentrations (8.20; 7.86), respectively. It is concluded that probiotic inclusion level had a significant effect on broiler growth responses, nutrient ADC, AME(n), and cecal microflora composition.

The aim of this work was to investigate the efficacy of a new multibacterial species probiotic in broiler nutrition. The probiotic contained 2 Lactobacillus strains, 1 Bifidobacterium strain, 1 Enterococcus strain, and 1 Pediococcus strain. Four hundred 1-d-old male Cobb broilers were allocated in 4 experimental treatments for 6 wk. The experimental treatments received a corn-soybean basal diet and were as follows: "control," with no other additions; "probiotic in feed and water," (PFW) with probiotic administered at 1 g/kg of feed for the whole period and in water on scheduled intervals during the first 4 wk; "probiotic in feed," (PF) with probiotic in feed as in PFW; and "antibiotic," (AB) with addition of avilamycin at 2.5 mg/kg of feed. Salinomycin Na was used as a coccidiostat. Each treatment had 5 replicates of 20 broilers. Treatment effects on parameters of broiler performance and cecal microbial ecology were determined. Broiler BW, feed intake, and feed conversion ratio were determined on a weekly and overall basis. Cecal microflora composition, concentration of volatile fatty acids, and activities of 5 bacterial glycolytic enzymes (alpha-galactosidase, beta-galactosidase, alpha-glucosidase, beta-glucosidase, and beta-glucuronidase) were determined at the end of the experiment. Overall, treatment PFW displayed a growth-promoting effect that did not differ from AB. Overall, feed conversion ratio in treatment AB was significantly better (P < or = 0.01) than the control treatment, whereas treatments PFW and PF were intermediate and not different from AB. Concentrations of bacteria belonging to Bifidobacterium spp., Lactobacillus spp., and gram-positive cocci were significantly (P < or = 0.05) higher in treatments PFW and PF compared with the control and AB treatments. Treatments PFW and PF had significantly higher specific activities of alpha-galactosidase and beta-galactosidase compared with the control and AB treatments. In conclusion, probiotic treatment PFW displayed a growth-promoting effect that was comparable to avilamycin treatment. In addition, treatments PFW and PF modulated the composition and, to an extent, the activities of the cecal microflora, resulting in a significant probiotic effect.