Thymol has been known as a functional phytochemical isolated from thyme essential oils and posses... more Thymol has been known as a functional phytochemical isolated from thyme essential oils and possesses antioxidant, antimicrobial, and anti-inflammatory properties. In this study, an in vitro lipopolysaccharide (LPS)-induced inflammation model using IPEC-J2 cell line was established to evaluate the inflammatory responses after thymol treatment. Cells were pre-treated with thymol for 1 h followed by LPS stimulation. Interleukin 8 (IL-8) secretion, reactive oxygen species (ROS) production, mRNA abundance of two pro-inflammatory cytokines, nutrient transporters, and tight junction proteins, transepithelial electrical resistance (TEER) and cell permeability were measured. The localization of zonula occludens-1 (ZO-1) and β-actin were also detected by immunofluorescent staining. The results showed that LPS stimulation increased IL-8 secretion, ROS production, and tumor necrosis factor alpha (TNF-α) mRNA abundance (P < 0.05), but the mRNA abundance of sodium-dependent glucose transporter 1 (SGLT1), excitatory amino acid transporter 1 (EAAC1) and H+/peptide cotransporter 1 (PepT1) were decreased (P < 0.05). However, thymol blocked ROS production (P < 0.05) and tended to decrease the production of LPS-induced IL-8 secretion (P = 0.0766). The mRNA abundance of IL-8 and TNF-α was reduced by thymol pre-treatment (P < 0.05), but thymol was unable to improve the gene expression of nutrient transporters (P > 0.05). TEER was reduced and cell permeability was increased after LPS stimulation (P < 0.05), but these effects were attenuated by thymol pre-treatment (P < 0.05). Moreover, thymol boosted ZO-1 and β-actin staining in the cells, but the mRNA abundance of ZO-1 and occludin-3 was not affected by either LPS or thymol treatments. These results indicated that thymol can enhance gut barrier structure and functions by reducing ROS production and pro-inflammatory cytokine gene expression in porcine epithelial cells during inflammation. The regulation of barrier function by thymol may be at post-transcriptional or post-translational levels.
Essential oils are defined as plant-derived natural bioactive compounds with positive effects on ... more Essential oils are defined as plant-derived natural bioactive compounds with positive effects on animal growth and health due to their antimicrobial and antioxidative properties. However, essential oils are very volatile, can evaporate rapidly and be rapidly absorbed in the upper gastrointestinal tract. In addition, due to their labile nature, the stability of essential oils during feed processing is often questionable, leading to variable final concentrations in feeds. Micro-encapsulation has become one of the most popular methods to deliver essential oils into the lower gut. The objective of the present study was double: 1) to validate and demonstrate the slow release of essential oils, such as thymol, micro-encapsulated in combination with organic acids in a matrix of triglycerides, in simulated swine gastric and intestinal fluids and 2) to evaluate the essential oil stability in the microparticles during feed pelleting process. In the in vitro release experiments, the microparticles were incubated in simulated gastric fluids for 2 hours and then the samples were incubated in simulated intestinal fluids for 0, 1, 2, 3, 4, 6, 8, 10, and 24 hours at 39°C. In the pelleting experiment, a wheat-corn basal diet with 2 kg of micro-encapsulated product was formulated and pelleted. The thymol content in the samples was analyzed by gas chromatography with flame-ionization detection. The results showed that 27.65% thymol was released in simulated gastric fluids and the rest of thymol was progressively released in intestinal fluids until completion, which was achieved by 24 hours. The thymol concentration was not significantly different between the mash feeds and pelleted feeds (P > 0.05). In conclusion, the micro-encapsulated organic acid and essential oil product was able to maintain the stability of thymol under a commercial pelleting process and allow a slow and progressive release of its active ingredients as thymol in simulated digestive fluids.
Sorghum has been increasingly used as animal feed ingredients due to low cost and high energy. Ho... more Sorghum has been increasingly used as animal feed ingredients due to low cost and high energy. However, anti-nutritional factors in sorghum negatively affect the availability of nutrients. This study aimed to mitigate the adverse impacts of sorghum on the growth performance of growing pigs by protease supplementation. Forty-two crossbred pigs (Duroc × Landrace) were randomly assigned into seven treatment groups: A (0% sorghum + 0% protease); B (20% sorghum + 0% protease); C (20% sorghum + 0.015% protease); D (50% sorghum + 0% protease); E (50% sorghum + 0.015% protease); F (80% sorghum + 0% protease); and G (80% sorghum + 0.015% protease). Growth performance, meat quality, serum biochemical parameters, and apparent ileal amino acids digestibility were measured. Results showed that average daily feed intake (ADFI) significantly increased in the group with 50% sorghum, and ADFI and average daily gain (ADG) increased in the group with 80% sorghum (P < 0.05). Precaval vein, mesenteric vein and portal vein serum concentration of histidine decreased (P < 0.05). The apparent ileal threonine digestibility increased with 20% sorghum and the apparent ileal lysine digestibility increased with 50% sorghum (P < 0.05). The apparent ileal digestibility of other amino acids decreased or remained stable with 80% sorghum. Protease supplementation improved feed conversion ratio (FCR) with 20% sorghum (P < 0.05) when compared to 20% sorghum without protease. The FCR, ADG, ADFI and meat quality of growing pigs with 50–80% dietary sorghum and protease supplementation were comparable to the control (P > 0.05). The apparent ileal digestibility of amino acids decreased in the group with an 80% sorghum amount except for lysine and valine. Sorghum could be a substitution for corn with a proper proportion (between 20% and 80%) of dietary feed. In conclusion, the supplementation of protease improved feed efficiency at 20% sorghum inclusion.
Feed ingredient prices and markets competitiveness prompt animal nutritionists to modulate nutrie... more Feed ingredient prices and markets competitiveness prompt animal nutritionists to modulate nutrients density of diets in order to control feed cost. By improving raw material digestibility, protease enzyme is an interesting tool to reduce costs and secure animal performance. This trial aimed to demonstrate usefulness of protease enzyme in piglets feeds by using it either on top or according to its matrix of valorization. A 42d pen study was conducted using 360 pigs (PIC) with an initial body weight (BW) of 6.15 kg equally distributed in 36 pens with 12 replications fed one of the following three treatments: CTRL) Control diet with ME of 3502/3386/3367 kcal/kg, Crude protein (CP) of 22.9/21.9/20.7 % for phases I (d0-7), II (d7-21), III (d21-42) respectively; OT) CTRL diet + 125 g/t Jefo Protease (Jefo, Canada) supplemented “On Top”; MT) CTRL diet reformulated to incorporate 125 g/t Jefo Protease according to its matrix of valorization. Data were subjected to statistical analyses of variance as a randomized complete block design. Differences among treatment means were determined with level of significance at P<0.05. Overall, pigs fed with protease on top (OT diets) showed significant (P<0.05) improvement on feed/gain (FG) with 1.273 versus 1.301 for CTRL. Pigs fed MT diets achieved similar growth performance to pigs fed CTRL diets but with cheaper feeds resulting in higher income over feed cost by comparison to pigs fed with CTRL diets ($42.87 vs $42.73 USD). In this trial, protease use with its matrix in a commercial pelleted diet accordingly reformulated in CP, AA and energy demonstrated potential to maintain BW and FG of pigs up to the level of classic diet and to allow economic savings on incomes over feed costs.
This study aimed to evaluate the effects of supplementing broiler diets with a dietary protease o... more This study aimed to evaluate the effects of supplementing broiler diets with a dietary protease on growth performance, digestive function, intestinal morphology, and meat quality as compared with feeding diets with or without an antibiotic growth promoter (AGP). A total of 240 1-day-old male chicks (Cobb 500, 48.3 ± 3.3 g) were distributed to three treatments with eight replicates (10 birds per replicate). Three treatments were: 1) corn–soybean meal basal diets (CTRL), 2) basal diets with 0.003% avilamycin (AB), and 3) basal diets with 0.0125% protease (PRT). The diets were provided as mash form, and birds were fed ad libitum during the whole experimental period. On day 45, birds were euthanized, and tissue and digesta samples were collected. On day 46, the remaining birds were processed in a commercial slaughterhouse, and breast muscle samples were collected. Despite a trend for a decreased feed conversion ratio (FCR) in the AB group during the whole phase (P = 0.071), no significant differences in growth performance parameters and relative weights of organs were observed (P > 0.05) among the groups. The AB and PRT groups showed significantly greater apparent ileal digestibility of amino acids (AA) compared with the CTRL group (P < 0.05). The PRT group significantly improved the morphology of duodenum and jejunum (P < 0.05). No differences were detected for meat quality, white striping, and woody breast among the groups (P > 0.05). For the gene expressions, the AB group showed a greater level of B0-system neutral amino acid co-transporter 1 and excitatory amino acid transporter 1 mRNA abundance compared with PRT group, while a significantly lesser level of cationic amino acid transporter 1 mRNA abundance was observed in the AB group compared with CTRL group (P < 0.05). The PRT group had a lesser level of peptide transporter 1 mRNA abundance in the jejunum than the CTRL group (P < 0.05). The highest mRNA abundances of zonula occludens-1 and cadherin 1 were observed in the CTRL group (P < 0.05). In conclusion, supplementation of avilamycin tended to reduce FCR and significantly improved AA utilization, and supplementation of dietary protease significantly enhanced intestinal morphology and AA utilization in broilers. In that respect, exogenous protease use appears to be an interesting tool to be considered in AGP reduction strategies.
Eugenol (4-allyl-2-methoxyphenol) is an essential oil component, possessing antimicrobial, anti-i... more Eugenol (4-allyl-2-methoxyphenol) is an essential oil component, possessing antimicrobial, anti-inflammatory, and antioxidative properties; however, the effect of eugenol on porcine gut inflammation has not yet been investigated. In this study, an in vitro lipopolysaccharide (LPS)-induced inflammation model in porcine intestinal epithelial cells (IPEC-J2) has been set up. Cells were pretreated with 100 μM (16.42 mg/L) eugenol for 2 h followed by 10 μg/mL LPS stimulation for 6 h. Proinflammatory cytokine secretion; reactive oxygen species; gene expression of proinflammatory cytokines, tight junction proteins, and nutrient transporters; the expression and distribution of zonula occludens-1 (ZO-1); transepithelial electrical resistance (TEER); and cell permeability were measured to investigate the effect of eugenol on inflammatory responses and gut barrier function. The results showed that eugenol pretreatment significantly suppressed the LPS-stimulated interleukin-8 level and the mRNA abundance of tumor necrosis factor-α and restored the LPS-stimulated decrease of the mRNA abundance of tight junction proteins, such as ZO-1 and occludin, and the mRNA abundance of nutrient transporters, such as B0 1 system ASC sodium-dependent neutral amino acid exchanger 2, sodium-dependent glucose transporter 1, excitatory amino acid transporter 1, and peptide transporter 1. In addition, eugenol improved the expression and even redistribution of ZO-1 and tended to increase TEER value and maintained the barrier integrity. In conclusion, a low dose of eugenol attenuated inflammatory responses and enhanced selectively permeable barrier function during LPS-induced inflammation in the IPEC-J2 cell line.
<p>Based on a subsample of 898 OTU; <i>p</i> values returned by Mothur; p value... more <p>Based on a subsample of 898 OTU; <i>p</i> values returned by Mothur; p values < 0.05 are in bold.</p><p>Beta-diversity analysis across chicken groups and according to <i>C</i>. <i>jejuni</i> colonization or feed additive use.</p
Thymol has been known as a functional phytochemical isolated from thyme essential oils and posses... more Thymol has been known as a functional phytochemical isolated from thyme essential oils and possesses antioxidant, antimicrobial, and anti-inflammatory properties. In this study, an in vitro lipopolysaccharide (LPS)-induced inflammation model using IPEC-J2 cell line was established to evaluate the inflammatory responses after thymol treatment. Cells were pre-treated with thymol for 1 h followed by LPS stimulation. Interleukin 8 (IL-8) secretion, reactive oxygen species (ROS) production, mRNA abundance of two pro-inflammatory cytokines, nutrient transporters, and tight junction proteins, transepithelial electrical resistance (TEER) and cell permeability were measured. The localization of zonula occludens-1 (ZO-1) and β-actin were also detected by immunofluorescent staining. The results showed that LPS stimulation increased IL-8 secretion, ROS production, and tumor necrosis factor alpha (TNF-α) mRNA abundance (P < 0.05), but the mRNA abundance of sodium-dependent glucose transporter 1 (SGLT1), excitatory amino acid transporter 1 (EAAC1) and H+/peptide cotransporter 1 (PepT1) were decreased (P < 0.05). However, thymol blocked ROS production (P < 0.05) and tended to decrease the production of LPS-induced IL-8 secretion (P = 0.0766). The mRNA abundance of IL-8 and TNF-α was reduced by thymol pre-treatment (P < 0.05), but thymol was unable to improve the gene expression of nutrient transporters (P > 0.05). TEER was reduced and cell permeability was increased after LPS stimulation (P < 0.05), but these effects were attenuated by thymol pre-treatment (P < 0.05). Moreover, thymol boosted ZO-1 and β-actin staining in the cells, but the mRNA abundance of ZO-1 and occludin-3 was not affected by either LPS or thymol treatments. These results indicated that thymol can enhance gut barrier structure and functions by reducing ROS production and pro-inflammatory cytokine gene expression in porcine epithelial cells during inflammation. The regulation of barrier function by thymol may be at post-transcriptional or post-translational levels.
Essential oils are defined as plant-derived natural bioactive compounds with positive effects on ... more Essential oils are defined as plant-derived natural bioactive compounds with positive effects on animal growth and health due to their antimicrobial and antioxidative properties. However, essential oils are very volatile, can evaporate rapidly and be rapidly absorbed in the upper gastrointestinal tract. In addition, due to their labile nature, the stability of essential oils during feed processing is often questionable, leading to variable final concentrations in feeds. Micro-encapsulation has become one of the most popular methods to deliver essential oils into the lower gut. The objective of the present study was double: 1) to validate and demonstrate the slow release of essential oils, such as thymol, micro-encapsulated in combination with organic acids in a matrix of triglycerides, in simulated swine gastric and intestinal fluids and 2) to evaluate the essential oil stability in the microparticles during feed pelleting process. In the in vitro release experiments, the microparticles were incubated in simulated gastric fluids for 2 hours and then the samples were incubated in simulated intestinal fluids for 0, 1, 2, 3, 4, 6, 8, 10, and 24 hours at 39°C. In the pelleting experiment, a wheat-corn basal diet with 2 kg of micro-encapsulated product was formulated and pelleted. The thymol content in the samples was analyzed by gas chromatography with flame-ionization detection. The results showed that 27.65% thymol was released in simulated gastric fluids and the rest of thymol was progressively released in intestinal fluids until completion, which was achieved by 24 hours. The thymol concentration was not significantly different between the mash feeds and pelleted feeds (P > 0.05). In conclusion, the micro-encapsulated organic acid and essential oil product was able to maintain the stability of thymol under a commercial pelleting process and allow a slow and progressive release of its active ingredients as thymol in simulated digestive fluids.
Sorghum has been increasingly used as animal feed ingredients due to low cost and high energy. Ho... more Sorghum has been increasingly used as animal feed ingredients due to low cost and high energy. However, anti-nutritional factors in sorghum negatively affect the availability of nutrients. This study aimed to mitigate the adverse impacts of sorghum on the growth performance of growing pigs by protease supplementation. Forty-two crossbred pigs (Duroc × Landrace) were randomly assigned into seven treatment groups: A (0% sorghum + 0% protease); B (20% sorghum + 0% protease); C (20% sorghum + 0.015% protease); D (50% sorghum + 0% protease); E (50% sorghum + 0.015% protease); F (80% sorghum + 0% protease); and G (80% sorghum + 0.015% protease). Growth performance, meat quality, serum biochemical parameters, and apparent ileal amino acids digestibility were measured. Results showed that average daily feed intake (ADFI) significantly increased in the group with 50% sorghum, and ADFI and average daily gain (ADG) increased in the group with 80% sorghum (P < 0.05). Precaval vein, mesenteric vein and portal vein serum concentration of histidine decreased (P < 0.05). The apparent ileal threonine digestibility increased with 20% sorghum and the apparent ileal lysine digestibility increased with 50% sorghum (P < 0.05). The apparent ileal digestibility of other amino acids decreased or remained stable with 80% sorghum. Protease supplementation improved feed conversion ratio (FCR) with 20% sorghum (P < 0.05) when compared to 20% sorghum without protease. The FCR, ADG, ADFI and meat quality of growing pigs with 50–80% dietary sorghum and protease supplementation were comparable to the control (P > 0.05). The apparent ileal digestibility of amino acids decreased in the group with an 80% sorghum amount except for lysine and valine. Sorghum could be a substitution for corn with a proper proportion (between 20% and 80%) of dietary feed. In conclusion, the supplementation of protease improved feed efficiency at 20% sorghum inclusion.
Feed ingredient prices and markets competitiveness prompt animal nutritionists to modulate nutrie... more Feed ingredient prices and markets competitiveness prompt animal nutritionists to modulate nutrients density of diets in order to control feed cost. By improving raw material digestibility, protease enzyme is an interesting tool to reduce costs and secure animal performance. This trial aimed to demonstrate usefulness of protease enzyme in piglets feeds by using it either on top or according to its matrix of valorization. A 42d pen study was conducted using 360 pigs (PIC) with an initial body weight (BW) of 6.15 kg equally distributed in 36 pens with 12 replications fed one of the following three treatments: CTRL) Control diet with ME of 3502/3386/3367 kcal/kg, Crude protein (CP) of 22.9/21.9/20.7 % for phases I (d0-7), II (d7-21), III (d21-42) respectively; OT) CTRL diet + 125 g/t Jefo Protease (Jefo, Canada) supplemented “On Top”; MT) CTRL diet reformulated to incorporate 125 g/t Jefo Protease according to its matrix of valorization. Data were subjected to statistical analyses of variance as a randomized complete block design. Differences among treatment means were determined with level of significance at P<0.05. Overall, pigs fed with protease on top (OT diets) showed significant (P<0.05) improvement on feed/gain (FG) with 1.273 versus 1.301 for CTRL. Pigs fed MT diets achieved similar growth performance to pigs fed CTRL diets but with cheaper feeds resulting in higher income over feed cost by comparison to pigs fed with CTRL diets ($42.87 vs $42.73 USD). In this trial, protease use with its matrix in a commercial pelleted diet accordingly reformulated in CP, AA and energy demonstrated potential to maintain BW and FG of pigs up to the level of classic diet and to allow economic savings on incomes over feed costs.
This study aimed to evaluate the effects of supplementing broiler diets with a dietary protease o... more This study aimed to evaluate the effects of supplementing broiler diets with a dietary protease on growth performance, digestive function, intestinal morphology, and meat quality as compared with feeding diets with or without an antibiotic growth promoter (AGP). A total of 240 1-day-old male chicks (Cobb 500, 48.3 ± 3.3 g) were distributed to three treatments with eight replicates (10 birds per replicate). Three treatments were: 1) corn–soybean meal basal diets (CTRL), 2) basal diets with 0.003% avilamycin (AB), and 3) basal diets with 0.0125% protease (PRT). The diets were provided as mash form, and birds were fed ad libitum during the whole experimental period. On day 45, birds were euthanized, and tissue and digesta samples were collected. On day 46, the remaining birds were processed in a commercial slaughterhouse, and breast muscle samples were collected. Despite a trend for a decreased feed conversion ratio (FCR) in the AB group during the whole phase (P = 0.071), no significant differences in growth performance parameters and relative weights of organs were observed (P > 0.05) among the groups. The AB and PRT groups showed significantly greater apparent ileal digestibility of amino acids (AA) compared with the CTRL group (P < 0.05). The PRT group significantly improved the morphology of duodenum and jejunum (P < 0.05). No differences were detected for meat quality, white striping, and woody breast among the groups (P > 0.05). For the gene expressions, the AB group showed a greater level of B0-system neutral amino acid co-transporter 1 and excitatory amino acid transporter 1 mRNA abundance compared with PRT group, while a significantly lesser level of cationic amino acid transporter 1 mRNA abundance was observed in the AB group compared with CTRL group (P < 0.05). The PRT group had a lesser level of peptide transporter 1 mRNA abundance in the jejunum than the CTRL group (P < 0.05). The highest mRNA abundances of zonula occludens-1 and cadherin 1 were observed in the CTRL group (P < 0.05). In conclusion, supplementation of avilamycin tended to reduce FCR and significantly improved AA utilization, and supplementation of dietary protease significantly enhanced intestinal morphology and AA utilization in broilers. In that respect, exogenous protease use appears to be an interesting tool to be considered in AGP reduction strategies.
Eugenol (4-allyl-2-methoxyphenol) is an essential oil component, possessing antimicrobial, anti-i... more Eugenol (4-allyl-2-methoxyphenol) is an essential oil component, possessing antimicrobial, anti-inflammatory, and antioxidative properties; however, the effect of eugenol on porcine gut inflammation has not yet been investigated. In this study, an in vitro lipopolysaccharide (LPS)-induced inflammation model in porcine intestinal epithelial cells (IPEC-J2) has been set up. Cells were pretreated with 100 μM (16.42 mg/L) eugenol for 2 h followed by 10 μg/mL LPS stimulation for 6 h. Proinflammatory cytokine secretion; reactive oxygen species; gene expression of proinflammatory cytokines, tight junction proteins, and nutrient transporters; the expression and distribution of zonula occludens-1 (ZO-1); transepithelial electrical resistance (TEER); and cell permeability were measured to investigate the effect of eugenol on inflammatory responses and gut barrier function. The results showed that eugenol pretreatment significantly suppressed the LPS-stimulated interleukin-8 level and the mRNA abundance of tumor necrosis factor-α and restored the LPS-stimulated decrease of the mRNA abundance of tight junction proteins, such as ZO-1 and occludin, and the mRNA abundance of nutrient transporters, such as B0 1 system ASC sodium-dependent neutral amino acid exchanger 2, sodium-dependent glucose transporter 1, excitatory amino acid transporter 1, and peptide transporter 1. In addition, eugenol improved the expression and even redistribution of ZO-1 and tended to increase TEER value and maintained the barrier integrity. In conclusion, a low dose of eugenol attenuated inflammatory responses and enhanced selectively permeable barrier function during LPS-induced inflammation in the IPEC-J2 cell line.
<p>Based on a subsample of 898 OTU; <i>p</i> values returned by Mothur; p value... more <p>Based on a subsample of 898 OTU; <i>p</i> values returned by Mothur; p values < 0.05 are in bold.</p><p>Beta-diversity analysis across chicken groups and according to <i>C</i>. <i>jejuni</i> colonization or feed additive use.</p
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