Protective effects of Lactobacillus on heat stress-induced intestinal injury in finisher broilers by regulating gut microbiota and stimulating epithelial development.

Heat stress (HS) is a critical challenge in broilers due to the high metabolic rate and lack of sweat glands. Results from this study show that implementing a cyclic chronic HS (34 °C for 7 h/d) to finisher broilers decreased the diversity of cecal microbiota and impaired intestinal barrier, resulting in gut leak and decreased body weight (both P < 0.05). These alterations might be related to inflammatory outbursts and the retarded proliferation of intestinal epithelial cells (IECs) according to the transcriptome analysis. Considering the potential beneficial properties of Lactobacillus on intestinal development and function, the protective effects of Lactobacillus rhamnosus (L. rhamnosus) on the intestine were investigated under HS conditions in this study. Orally supplemented L. rhamnosus improved the composition of cecal microbiota and upregulated the transcription of tight junction proteins in both duodenum and jejunum, with a consequent suppression in intestinal gene expressions of pro-inflammatory cytokines and facilitation in digestive capability. Meanwhile, the jejunal villus height of the birds that received L. rhamnosus was significantly higher compared with those treated with the broth (P < 0.05). The expression abundances of genes related to IECs proliferation and differentiation were increased by L. rhamnosus, along with upregulated mRNA levels of Wnt3a and ß-catenin in jejunum. In addition, L. rhamnosus attenuated enterocyte apoptosis as indicated by decreased caspase-3 and caspase-9 gene expressions. The results indicated that oral administration with L. rhamnosus mitigated HS-induced dysfunction by promoting intestinal development and epithelial maturation in broilers and that the effects of L. rhamnosus might be dependent of Wnt/ß-catenin signaling.

Effect of supplemental methyl sulfonyl methane on performance, carcass and meat quality and oxidative status in chronic cyclic heat-stressed finishing broilers.

Methyl sulfonyl methane (MSM) is available as a dietary supplement for human and has been associated with multiple health benefits such as reduction of oxidative stress. Heat stress (HS) is an environmental stressor challenging poultry production and known to inflict oxidative stress. We hypothesized that dietary MSM could attenuate HS-induced detrimental effects in broilers mediated by enhancement of antioxidant defenses. Hence, seven hundred ninety-two 1-day-old male Ross 308 broilers were allocated to 3 dietary treatments composed of corn-soybean meal diets with 0 (Ctrl), 1, or 2 g/kg MSM, with 12 replicates (22 birds each) per treatment for 39 d and subjected to a chronic cyclic HS model (temperature of 34°C and 52-58% relative humidity for 6 h daily) from d 24 to 39. MSM at 1 and 2 g/kg linearly increased daily gain and decreased feed-to-gain ratio compared with Ctrl in the grower phase (d 10-21, both P < 0.05). In the finisher phase (d 21-39) none of the performance and carcass indices were affected by treatment (P > 0.05). Nonetheless, data suggest reduced mortality by feeding MSM during HS. Also, during HS the diets with graded levels of MSM resulted in reduced rectal temperatures (P < 0.05) along with linearly decreased panting frequency on d 24 (P < 0.05). MSM supplemented birds showed a trend for linearly decreased thiobarbituric acid reactive substances of breast meat upon simulated retail display (P = 0.078). In addition, MSM administration linearly decreased lipid oxidation in plasma (d 25 and 39, P < 0.05) and breast muscle at d 23 (P < 0.05), concomitantly with linearly increased glutathione levels in erythrocytes (d 23 and 39, P < 0.05; d 25, P < 0.1) and breast muscle (d 23, P < 0.05; d 39, P < 0.1). In conclusion, MSM increased growth performance of broilers during grower phase, and exhibited positive effects on heat tolerance mediated by improved antioxidant capacity in broilers resulting in lower mortality in finisher phase.

Calcium supplementation in low nutrient density diet for meat ducks improves breast meat tenderness associated with myocyte apoptosis and proteolytic changes.

To define the relationship between dietary nutrient density, calcium (Ca), and meat quality in meat ducks. A total of 288 male Cherry Valley SM3 medium ducklings were fed a common standard starter diet until d 14. At 15 d of age, ducks were randomly divided into 2 treatment groups and fed either a conventional diet or a low nutrient density (LND) diet. Compared with the conventional diet, the energy was reduced in the LND diet by 8.6% and 16.8% in grower (15 to 35 d) and finisher (36 to 56 d) phases, respectively, while other essential nutrients were kept proportionate to energy. The LND diet decreased the shear force (P < 0.05) and increased the lightness values of the pectoralis muscle when compared to the conventional diet, suggesting that LND diet exerted a beneficial role in meat quality. Subsequently, the effects of grated Ca in the LND diet on meat quality of pectoralis muscle were evaluated. A total of 576 male ducklings were fed a common starter diet until d 14, followed by feeding 4 LND diets with 0.5%, 0.7%, 0.9%, and 1.1% Ca. The results show that LND diets with 0.7% or more Ca decreased the shear force of pectoralis major muscle in 42-d-old meat ducks (P < 0.05). To explore the mechanism underlying Ca and tenderness, data from birds fed either 0.5% or 1.1% Ca in the LND diet indicated that birds fed 1.1% Ca exhibited lower shear force, upregulated calpains 1 expression, and higher calpains activity compared to those fed the LND diet with 0.5% Ca (P < 0.05). Moreover, the 1.1% Ca LND diet induced a higher myocyte apoptosis (P = 0.06) and upregulated mRNA expression of caspase-3 (P = 0.07) in breast muscle. Our data suggest that LND diets with 0.9% or 1.1% Ca had a positive role in the tenderness of breast meat, particularly the enhancing effect of 1.1% Ca LND diet on tenderness seems to be associated with proteolytic changes of myofibrillar proteins and myocyte apoptosis in meat ducks.

The Effect of Amino Acids on Production of SCFA and bCFA by Members of the Porcine Colonic Microbiota.

Functional amino acids supplementation to farm animals is considered to not only be beneficial by regulating intestinal barrier, oxidative stress, and immunity, but potentially also by impacting the gut microbiota. The impact of amino acids on a piglet-derived colonic microbiota was evaluated using a 48-h in vitro batch incubation strategy. The combination of 16S rRNA gene profiling with flow cytometry demonstrated that specific microbial taxa were involved in the fermentation of each of the amino acids resulting in the production of specific metabolites. Branched chain amino acids (leucine, isoleucine, valine) strongly increased branched-chain fatty acids (+23.0 mM) and valerate levels (+3.0 mM), coincided with a marked increase of Peptostreptococcaceae. Further, glutamine and glutamate specifically stimulated acetate (~20 mM) and butyrate (~10 mM) production, relating to a stimulation of a range of families containing known butyrate-producing species (Ruminococcaceae, Oscillospiraceae, and Christensenellaceae). Finally, while tryptophan was only fermented to a minor extent, arginine and lysine specifically increased propionate levels (~2 mM), likely produced by Muribaculaceae members. Overall, amino acids were thus shown to be selectively utilized by microbes originating from the porcine colonic microbiota, resulting in the production of health-related short-chain fatty acids, thus confirming the prebiotic potential of specific functional amino acids.

Dietary Resistant Starch From Potato Regulates Bone Mass by Modulating Gut Microbiota and Concomitant Short-Chain Fatty Acids Production in Meat Ducks.

Gut microbiota interfered with using prebiotics may improve bone mass and alleviate the onset of bone problems. This study aimed to investigate the beneficial effect of resistant starch from raw potato starch (RPS) on bone health in meat ducks. Response to the dietary graded level of RPS supplementation, both tibia strength and ash were taken out linear and quadratic increase and positively correlated with increased propionate and butyrate levels in cecal content. Moreover, further outcomes of gut microbiota and micro-CT analysis showed the beneficial effect of RPS on bone mass might be associated with higher Firmicutes proportion and the production of short-chain fatty acids (SCFAs) in the cecum. Consistent with improving bone mass, SCFAs promoted phosphorus absorption, decreased the digestive tract pH, and enhanced intestinal integrity, which decreased the expression of pro-inflammatory genes in both gut and bone marrow, and consequently depressed osteoclastic bone resorption mediated by inflammatory cytokines. These findings highlight the importance of the "gut-bone" axis and provide new insight into the effect of prebiotics on bone health.

Acidification of drinking water improved tibia mass of broilers through the alterations of intestinal barrier and microbiota.

OBJECTIVE: Diet acidification supplementation is known to influence intestinal morphology, gut microbiota, and on phosphorus (P) utilization of broilers. Alterations in intestinal barrier and microbiota have been associated with systemic inflammation and thus regulating bone turnover. Hence the effect of acidifier addition to drinking water on tibia mass and the linkages between intestinal integrity and bone were studied. METHODS: One-d-old male broilers were randomly assigned to normal water (control) or continuous supply of acidified water (2% the blend of 2-hydroxy-4-methylthiobutyric acid, lactic, and phosphoric acid) group with 5 replicates of 10 chicks per replicate for 42 d. RESULTS: Acidification of drinking water improved the ash percentage and calcium content of tibia at 42 d. Broilers receiving acidified water had increased serum P concentration compared to control birds. The acidified group showed improved intestinal barrier, evidenced by increased wall thickness, villus height, the villus height to crypt depth ratio, and upregulated mucin-2 expression in ileum. Broilers receiving drinking water containing mixed organic acids had a higher proportion of Firmicutes and the ratio of Firmicutes and Bacteroidetes, as well as a lower population of Proteobacteria. Meanwhile, the addition of acidifier to drinking water resulted in declined ileal and serum proinflammatory factors level and increased immunoglobulin concentrations in serum. Concerning bone remodeling, acidifier addition was linked to a decrease in serum C-terminal cross-linked telopeptide of type I collagen and tartrate-resistant acid phosphatase reflecting bone resorption, whereas it did not apparently change serum alkaline phosphatase activity that is a bone formation marker. CONCLUSION: Acidified drinking water increased tibia mineral deposition of broilers, which was probably linked with higher P utilization and decreased bone resorption through improved intestinal integrity and gut microbiota and through decreased systemic inflammation.

25-hydroxycholecalciferol reverses heat induced alterations in bone quality in finisher broilers associated with effects on intestinal integrity and inflammation.

BACKGROUND: Alterations in ambient temperature have been associated with multiple detrimental effects on broilers such as intestinal barrier disruption and dysbiosis resulting in systemic inflammation. Inflammation and 25-hydroxycholecalciferol (25-OH-D3) have shown to play a negative and positive role, respectively, in the regulation of bone mass. Hence the potential of 25-OH-D3 in alleviating heat induced bone alterations and its mechanisms was studied. RESULTS: Heat stress (HS) directly induced a decrease in tibia material properties and bone mass, as demonstrated by lower mineral content, and HS caused a notable increase in intestinal permeability. Treatment with dietary 25-OH-D3 reversed the HS-induced bone loss and barrier leak. Broilers suffering from HS exhibited dysbiosis and increased expression of inflammatory cytokines in the ileum and bone marrow, as well as increased osteoclast number and activity. The changes were prevented by dietary 25-OH-D3 administration. Specifically, dietary 25-OH-D3 addition decreased abundance of B- and T-cells in blood, and the expression of inflammatory cytokines, especially TNF-α, in both the ileum and bone marrow, but did not alter the diversity and population or composition of major bacterial phyla. With regard to bone remodeling, dietary 25-OH-D3 supplementation was linked to a decrease in serum C-terminal cross-linked telopeptide of type I collagen reflecting bone resorption and a concomitant decrement in osteoclast-specific marker genes expression (e.g. cathepsin K), whereas it did not apparently change serum bone formation markers during HS. CONCLUSIONS: These data underscore the damage of HS to intestinal integrity and bone health, as well as that dietary 25-OH-D3 supplementation was identified as a potential therapy for preventing these adverse effects.

Production of selenium-enriched microalgae as potential feed supplement in high-rate algae ponds treating domestic wastewater.

This study assessed the selenium (Se) removal efficiency of two pilot-scale high-rate algae ponds (HRAPs) treating domestic wastewater and investigated the production of Se-enriched microalgae as potential feed supplement. The HRAP-Se had an average Se, NH4+-N, total phosphorus and COD removal efficiency of, respectively, 43%, 93%, 77%, and 70%. Inorganic Se taken up by the microalgae was mainly (91%) transformed to selenoamino acids, and 49-63% of Se in the Se-enriched microalgae was bioaccessible for animals. The crude protein content (48%) of the microalgae was higher than that of soybeans, whereas the essential amino acid content was comparable. Selenium may induce the production of the polyunsaturated fatty acids omega-3 and omega-6 in microalgae. Overall, the production of Se-enriched microalgae in HRAPs may offer a promising alternative for upgrading low-value resources into high-value feed supplements, supporting the drive to a circular economy.

Effect of vitamin E level and dietary zinc source on performance and intestinal health parameters in male broilers exposed to a temperature challenge in the finisher period.

The objective of this study was to evaluate the interaction of zinc source (ZnSO4 vs. zinc amino acid complex) and vitamin E level (50 IU vs. 100 IU) on performance and intestinal health of broilers exposed to a temperature challenge in the finisher period. A total of 1224 day old male Ross 308 broilers were randomly distributed among 4 dietary treatments (9 replicates per treatment). Dietary treatments were organized in a 2 × 2 factorial arrangement: two sources of zinc, 60 mg/kg of Zn as ZnSO4 .7H2 O or 60 mg/kg of Zn as zinc amino acid complexes (ZnAA) combined with two levels of vitamin E (50 or 100 IU/kg). Zinc and vitamin E were added to a wheat/rye-based diet that was designed to create a mild nutritional challenge. From day 28 until day 36 (finisher period), all birds were subjected to chronic cyclic high temperatures (32°C ± 2°C and RH 55-65% for 6 h daily). The combination of ZnAA and 50 IU/kg of vitamin E improved weight gain in the starter (day 0-10), finisher (day 28-36) and overall period (day 0-36) and feed conversion ratio in the starter (day 0-10) and finisher phase (day 28-36). Providing Zn as ZnAA significantly improved villus length and villus/crypt ratio in the starter, grower and finisher period and decreased infiltration of T-lymphocytes and ovotransferrin leakage in the finisher period. In conclusion, providing broilers with a diet supplemented with ZnAA and a vitamin E level of 50 IU/kg, resulted in better growth performance as compared to all other dietary treatments. Interestingly, under the conditions of this study, positive effects of ZnAA on performance did not occur when vitamin E was supplemented at 100 IU/kg in feed. Moreover, providing zinc as zinc amino acid complex improved intestinal health.

Short-chain fructo-oligosaccharides supplementation to suckling piglets: Assessment of pre- and post-weaning performance and gut health.

Farmers face difficulties in redeeming their investment in larger litter sizes since this comes with larger litter heterogenicity, lower litter resilience and risk of higher mortality. Dietary oligosaccharides, given to the sow, proved beneficial for the offspring's performance. However, giving oligosaccharides to the suckling piglet is poorly explored. Therefore, this field trial studied the effect of dietary short-chain fructo-oligosaccharides (scFOS; 1g/day; drenched) supplementation to low (LBW, lower quartile), normal (NBW, two intermediate quartiles) and high (HBW, upper quartile) birth weight piglets from birth until 7 or 21 days of age. Performance parameters, gut microbiome and short-chain fatty acids profile of feces and digesta were assessed at birth (d 0), d 7, weaning (d 21.5) and 2 weeks post-weaning (d 36.5). Additional parameters reflecting gut health (intestinal integrity and morphology, mucosal immune system) were analysed at d 36.5. Most parameters changed with age or differed with the piglet's birth weight. Drenching with scFOS increased body weight by 1 kg in NBW suckling piglets and reduced the post-weaning mortality rate by a 100%. No clear difference in the IgG level, the microbiota composition and fermentative activity between the treatment groups was observed. Additionnally, intestinal integrity, determined by measuring intestinal permeability and regenerative capacity, was similar between the treatment groups. Also, intestinal architecture (villus lenght, crypt depth) was not affected by scFOS supplementation. The density of intra-epithelial lymphocytes and the expression profiles (real-time qPCR) for immune system-related genes (IL-10, IL-1ß, IL-6, TNFα and IFNγ) were used to assess mucosal immunity. Only IFNγ expression, was upregulated in piglets that received scFOS for 7 days. The improved body weight and the reduced post-weaning mortality seen in piglets supplemented with scFOS support the view that scFOS positively impact piglet's health and resilience. However, the modes of action for these effects are not yet fully elucidated and its potential to improve other performance parameters needs further investigation.

Impact of Dietary Manganese on Intestinal Barrier and Inflammatory Response in Broilers Challenged with Salmonella Typhimurium.

Growing concern for public health and food safety has prompted a special interest in developing nutritional strategies for removing waterborne and foodborne pathogens, including Salmonella. Strong links between manganese (Mn) and intestinal barrier or immune function hint that dietary Mn supplementation is likely to be a promising approach to limit the loads of pathogens in broilers. Here, we provide evidence that Salmonella Typhimurium (S. Typhimurium, 4 × 108 CFUs) challenge-induced intestinal injury along with systemic Mn redistribution in broilers. Further examining of the effect of dietary Mn treatments (a basal diet plus additional 0, 40, or 100 mg Mn/kg for corresponding to Mn-deficient, control, or Mn-surfeit diet, respectively) on intestinal barrier and inflammation status of broilers infected with S. Typhimurium revealed that birds fed the control and Mn-surfeit diets exhibited improved intestinal tight junctions and microbiota composition. Even without Salmonella infection, dietary Mn deficiency alone increased intestinal permeability by impairing intestinal tight junctions. In addition, when fed the control and Mn-surfeit diets, birds showed decreased Salmonella burdens in cecal content and spleen, with a concomitant increase in inflammatory cytokine levels in spleen. Furthermore, the dietary Mn-supplementation-mediated induction of cytokine production was probably associated with the nuclear factor kappa-B (NF-κB)/hydrogen peroxide (H2O2) pathway, as judged by the enhanced manganese superoxide dismutase activity and the increased H2O2 level in mitochondria, together with the increased mRNA level of NF-κB in spleen. Ingenuity-pathway analysis indicated that acute-phase response pathways, T helper type 1 pathway, and dendritic cell maturation were significantly activated by the dietary Mn supplementation. Our data suggest that dietary Mn supplementation could enhance intestinal barrier and splenic inflammatory response to fight against Salmonella infection in broilers.

Effect of zinc oxide sources and dosages on gut microbiota and integrity of weaned piglets.

Zinc oxide (ZnO) supplied at pharmacological dosage in diets of weaned piglets improves growth performance. However, it causes environmental contamination and induces bacterial antibiotic resistance, yet this practice is debated. The effects on gut microbiota and integrity in weaned piglets of conventional ZnO at nutritional and pharmacological dosage (110 and 2,400 mg/kg Zn, respectively) were compared to an alternative ZnO source at 110 and 220 mg/kg Zn. Each of the four treatments was applied to four pens (two piglets/pen; weaning age, 20 days) for 15 days, and piglets were sampled on day 15 to determine indices of gut integrity. Feeding conventional ZnO at 2,400 mg/kg Zn reduced coliforms and Escherichia coli in distal small intestine as compared to conventional ZnO at 110 mg/kg (-1.7 and -1.4 log10 cfu/g, respectively), whereas the alternative ZnO reduced only coliforms, irrespective of dosage (-1.6 to -1.7 log10 cfu/g). Transepithelial electrical resistance of distal small intestinal mucosa was higher for pigs fed the alternative ZnO source as compared with groups fed 110 mg/kg Zn of conventional ZnO, in line with a trend for higher gene expression of claudin-1 and zona occludens-1. Interestingly, the alternative ZnO source at 110 and 220 mg/kg Zn increased intestinal alkaline phosphatase gene transcript as compared to conventional ZnO at 110 mg/kg Zn, whereas the alternative ZnO source at 110 mg/kg Zn exhibited higher Zn concentrations in mucosa (2,520 µg/g) as compared to conventional ZnO at 110 mg/kg Zn (1,211 µg/g). However, assessing alkaline phosphatase activity, no significant effects were found. In conclusion, the alternative ZnO reduced digesta Enterobacteriaceae numbers and improved gut integrity, albeit similar or better, depending on the dosage, to the effects of pharmacological dosage of conventional ZnO.

Effect of organic acids on Salmonella colonization and shedding in weaned piglets in a seeder model.

Piglets (n = 128) weaned at 21 days of age were used in a 35-day seeder model to evaluate the effects of dietary additives differing in active ingredients, chemical, and physical formulation, and dose on Salmonella colonization and shedding and intestinal microbial populations. Treatments were a negative control (basal diet), the positive control (challenged, basal diet), and six treatments similar to the positive control but supplemented with the following active ingredients (dose excluding essential oils or natural extracts): triglycerides with butyric acid (1.30 g kg(-1)); formic and citric acids and essential oils (2.44 g kg(-1)); coated formic, coated sorbic, and benzoic acids (2.70 g kg(-1)); salts of formic, sorbic, acetic, and propionic acids, their free acids, and natural extracts (2.92 g kg(-1)); triglycerides with caproic and caprylic acids and coated oregano oil (1.80 g kg(-1)); and caproic, caprylic, lauric, and lactic acids (1.91 g kg(-1)). On day 6, half the piglets (seeder pigs) in each group were orally challenged with a Salmonella Typhimurium nalidixic acid-resistant strain (4 × 10(9) and 1.2 × 10(9) log CFU per pig in replicate experiments 1 and 2, respectively). Two days later, they were transferred to pens with an equal number of contact pigs. Salmonella shedding was determined 2 days after challenge exposure and then on a weekly basis. On day 34 or 35, piglets were euthanized to sample tonsils, ileocecal lymph nodes, and ileal and cecal digesta contents. The two additives, both containing short-chain fatty acids and one of them also containing benzoic acid and the other one also containing essential oils, and supplemented at more than 2.70 g kg(-1), showed evidence of reducing Salmonella fecal shedding and numbers of coliforms and Salmonella in cecal digesta. However, colonization of tonsils and ileocecal lymph nodes by Salmonella was not affected. Supplementing butyric acid and medium-chain fatty acids at the applied dose failed to inhibit Salmonella contamination in the current experimental setup.