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Xochitl Hernandezvelasco - One of the best experts on this subject based on the ideXlab platform.
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effect of bacillus direct fed microbial on leaky gut serum peptide yy concentration Bone Mineralization and ammonia excretion in neonatal female turkey poults fed with a rye based diet
Poultry Science, 2020Co-Authors: G Tellez, Juan D Latorre, Xochitl Hernandezvelasco, B M Hargis, Ruben Merinoguzman, Margarita A Arreguinnava, J A Maguey, M A Michel, P A Moore, Guillermo TellezisaiasAbstract:Rye is high in nonstarch polysaccharides (NSP), a complex carbohydrate which cannot be digested by poultry as they lack the endogenous enzymes to do so. Exogenous carbohydrases must therefore be supplemented to avoid the antinutritional effects associated with a high NSP diet. The objectives of the present study were to evaluate the effects of a rye-based diet with and without supplementation of a Bacillus direct-fed microbial (DFM) on body weight, Bone Mineralization, and leaky gut, as well as its role on influencing serum concentrations of peptide YY (PPY) and the ammonia concentration in turkey manure. Two independent trials were conducted. In each experiment, day-of-hatch female turkey poults were neck tagged and randomly assigned to either a control rye-based diet or a rye-based diet supplemented with the DFM (n = 25 birds/group). At 10 days-of-age, poults in both groups were administered with an appropriate dose of fluorescein isothiocyanate-dextran (FITC-d) by oral gavage. One hour later, all poults were euthanized. Blood was collected to evaluate serum FITC-d and PPY concentrations. Furthermore, in Trial 2 only, both tibias were removed for assessment of Bone parameters, and turkey manure was collected to evaluate physicochemical analysis. In both trials, poults treated with the DFM showed a significant increase (P < 0.05) in body weight and body weight gain as compared with control nontreated poults. Poults that received the DFM also had a significant reduction in serum levels of PPY and FITC-d when compared with control nontreated poults. In Trial 2, turkeys treated with the DFM had a substantial increase in tibia strength, tibia diameter, total ash, calcium, and phosphorus when compared with control nontreated turkeys. Their manure was also shown to have a significant reduction in the concentration of ammonia. This is the first report of a commercial DFM reducing the concentration of this compound in turkey manure. In summary, the results of the present study confirm that turkeys fed with a rye-based diet have a significant increase in gut permeability, a reduced body weight, and decreased Bone Mineralization when compared with turkeys fed with the DFM. Turkeys that received the rye-based diet supplemented with the Bacillus-DFM also had a significant reduction in the serum concentration of PPY when compared with control turkeys. This finding suggests a possible prebiotic effect of rye, warranting future studies to test this effect. Further studies to evaluate the microbiota diversity, as well as the concentration of ceca short-chain fatty acids, are also necessary to confirm the reliability of PPY as a potential metabolomic biomarker in poultry.
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assessment of a nutritional rehabilitation model in two modern broilers and their jungle fowl ancestor a model for better understanding childhood undernutrition
Frontiers in Nutrition, 2018Co-Authors: Mikayla F A Baxter, Juan D Latorre, Sami Dridi, Dawn A Koltes, Elizabeth S Greene, Stephen W Bickler, Jae H Kim, Ruben Merinoguzman, Xochitl HernandezvelascoAbstract:This article is the first in a series of manuscripts to evaluate nutritional rehabilitation in chickens as a model to study interventions in children malnutrition (Part 1: Performance, Bone Mineralization, and Intestinal Morphometric Analysis). Inclusion of rye in poultry diets induces a nutritional deficit that leads to increased bacterial translocation, intestinal viscosity, and decreased Bone Mineralization. However, it is unclear the effect of diet on developmental stage or genetic strain. Therefore, the objective was to determine the effects of a rye diet during either the early or late phase of development on performance, Bone Mineralization, and intestinal morphology across three diverse genetic backgrounds. Modern 2015 (Cobb 500) broiler chicken, 1995 Cobb broiler chicken, and the Giant Jungle Fowl were randomly allocated into four different dietary treatments. Dietary treatments were (1) a control corn-based diet throughout the trial (corn-corn); (2) an early phase malnutrition diet where chicks received a rye-based diet for 10 days, and then switched to the control diet (rye-corn); (3) a malnutrition rye-diet that was fed throughout the trial (rye-rye); and (4) a late phase malnutrition diet where chicks received the control diet for 10 days, and then switched to the rye diet for the last phase (corn-rye). At 10 days of age, chicks were weighed and diets were switched in groups 2 and 4. At day 20 of age, all chickens were weighed and euthanized to collect Bone and intestinal samples. Body weight, weight gain, and Bone Mineralization were different across diet, genetic line, age and all two- and three-way interactions (P < 0.05). Overall, Jungle Fowl were the most tolerant to a rye-based diet, and both the modern and 1995 broilers were significantly affected by the high rye-based diet. However, the 1995 broilers consuming the rye-based diet appeared to experience more permanent effects when compared with the modern broiler. The results of this study suggest that chickens have a great potential as a nutritional rehabilitation model in human trials. The 1995 broilers line was an intermediate genetic line between the fast growing modern line and the non-selected Jungle Fowl line, suggesting that it would be the most appropriate model to study for future studies.
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rye affects bacterial translocation intestinal viscosity microbiota composition and Bone Mineralization in turkey poults
PLOS ONE, 2015Co-Authors: G Tellez, Juan D Latorre, B M Hargis, V A Kuttappan, Xochitl HernandezvelascoAbstract:Previously, we have reported that rye significantly increased both viscosity and Clostridium perfringens proliferation when compared with corn in an in vitro digestive model. Two independent trials were conducted to evaluate the effect of rye as a source of energy on bacterial translocation, intestinal viscosity, gut microbiota composition, and Bone Mineralization, when compared with corn in turkey poults. In each experiment, day-of-hatch, turkey poults were randomly assigned to either a corn or a rye diet (n = 0 /group). At 10 d of age, in both experiments, 12 birds/group were given an oral gavage dose of fluorescein isothiocyanate dextran (FITC-d). After 2.5 h of oral gavage, blood and liver samples were collected to evaluate the passage of FITC-d and bacterial translocation (BT) respectively. Duodenum, ileum and cecum gut sections were collected to evaluate intestinal viscosity and to enumerate gut microbiota. Tibias were collected for observation of Bone parameters. Broilers fed with a rye diet showed increased (p<0.05) intestinal viscosity, BT, and serum FITC-d. Bacterial enumeration revealed that turkey poults fed with rye had increased the number of total lactic acid bacteria (LAB) in all three sections of the gastrointestinal tract evaluated when compared to turkey poults fed with corn. Turkey poults fed with rye also had significantly higher coliforms in duodenum and ileum but not in the ceca, whereas the total number of anaerobes increased only in duodenum. A significant reduction in Bone strength and Bone Mineralization was observed in turkey poults fed with rye when compared with corn fed turkey poults. In conclusion, rye evoked mucosal damage in turkey poults that increased intestinal viscosity, increased leakage through the intestinal tract, and altered the microbiota composition and Bone Mineralization. Studies to evaluate dietary inclusion of selected Direct-Fed Microbial (DFM) candidates that produce exogenous enzymes in rye fed turkey poults are currently being evaluated.
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role of a bacillus subtilis direct fed microbial on digesta viscosity bacterial translocation and Bone Mineralization in turkey poults fed with a rye based diet
Frontiers in Veterinary Science, 2014Co-Authors: Juan D Latorre, Xochitl Hernandezvelasco, Michael H Kogut, J L Vicente, Ross Wolfenden, A D Wolfenden, B M Hargis, V A Kuttappan, G TellezAbstract:Rye contains high concentrations of non-starch polysaccharides (NSP), leading to reduced digestibility. Since poultry have little or no endogenous enzymes capable of hydrolyzing these NSP, exogenous carbohydrases as feed additives are used in an attempt to reduce the anti-nutritional effects of these polysaccharides. Previously, an in vitro study conducted in our laboratory showed that inclusion of certain Bacillus direct-fed microbial (DFM) candidates that produce exogenous phytase, lipase, protease, cellulase and xylanase in high NSP diets significantly reduced both digesta viscosity and Clostridium perfringens proliferation. In the present study, rye-based turkey starter diets with or without Bacillus-DFM were administered ad libitum to day-of-hatch turkey poults in two independent experiments. In both experiments, day-of-hatch turkey poults were randomly assigned to either a control diet (CON) or a DFM treated diet (n = 25 birds/group). At ten days–of-age, all turkey poults from experiments 1 and 2 were weighted and 12 turkey poults/group were randomly selected and humanely killed. Liver samples were aseptically collected to evaluate bacterial translocation, and intestinal digesta samples were individually collected to evaluate viscosity. Additionally, in experiment 2 both tibias were removed for assessment of Bone parameters. In both experiments, the TRT group showed a reduction in the total number of coliforms in the liver as well as a reduced digesta viscosity when compared to the CON group (P<0.05). Turkey poults fed the Bacillus-DFM candidate had increased tibia diameter, breaking strength, ash content, calcium content, and phosphorus content when compared with CON turkey poults. In summary, turkey poults fed with a rye-based diet without DFM showed an increase in bacterial translocation and digesta viscosity, accompanied by a reduction in Bone Mineralization; however these adverse effects can be prevented by the inclusion of selected a Bacillus-DFM candida
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utilization of rye as energy source affects bacterial translocation intestinal viscosity microbiota composition and Bone Mineralization in broiler chickens
Frontiers in Genetics, 2014Co-Authors: G Tellez, Juan D Latorre, Xochitl Hernandezvelasco, Michael H Kogut, B M Hargis, V A Kuttappan, Amanda Wolfenden, Walter Bottje, L R Bielke, Olivia B FaulknerAbstract:Two independent trials were conducted to evaluate the utilization of rye as energy source on bacterial translocation, intestinal viscosity, gut integrity, gut microbiota composition, and Bone Mineralization, when compared with a traditional cereal (corn) in broiler chickens. In each experiment, day-of-hatch, broiler chickens were randomly assigned to either a corn or a rye diet (n = 20 chickens/group). At 10d of age, in both experiments, 12 chickens/group were randomly selected, and given an oral gavage dose of fluorescein isothiocyanate dextran (FITC-d). After 2.5 h of oral gavage, blood samples were collected to determine the passage of FITC-d. The liver was collected from each bird to evaluate bacterial translocation (BT). Duodenum, ileum and cecum gut sections were collected to evaluate intestinal viscosity and to enumerate gut microbiota. Tibias were collected for observation of Bone parameters. Broilers fed with rye showed increased (p < 0.05) intestinal viscosity, BT, and serum FITC-d. Bacterial enumeration revealed that chickens fed with rye had increased the number of total lactic acid bacteria (LAB) in all three sections of the gastrointestinal tract evaluated when compared to chickens fed with corn. Chickens fed with rye also had significantly higher coliforms in duodenum and ileum, whereas the total number of anaerobes increased only in duodenum. A significant reduction in Bone strength and Bone Mineralization was observed in chickens fed with rye when compared with corn fed chickens. In conclusion, rye evoked mucosal damage in chickens that alter the intestinal viscosity, increased leakage through the intestinal tract, and altered the microbiota composition as well as Bone Mineralization. Studies to evaluate dietary inclusion of selected DFM candidates that produce exogenous enzymes in rye fed chickens are currently being evaluated.
G Tellez - One of the best experts on this subject based on the ideXlab platform.
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effect of bacillus direct fed microbial on leaky gut serum peptide yy concentration Bone Mineralization and ammonia excretion in neonatal female turkey poults fed with a rye based diet
Poultry Science, 2020Co-Authors: G Tellez, Juan D Latorre, Xochitl Hernandezvelasco, B M Hargis, Ruben Merinoguzman, Margarita A Arreguinnava, J A Maguey, M A Michel, P A Moore, Guillermo TellezisaiasAbstract:Rye is high in nonstarch polysaccharides (NSP), a complex carbohydrate which cannot be digested by poultry as they lack the endogenous enzymes to do so. Exogenous carbohydrases must therefore be supplemented to avoid the antinutritional effects associated with a high NSP diet. The objectives of the present study were to evaluate the effects of a rye-based diet with and without supplementation of a Bacillus direct-fed microbial (DFM) on body weight, Bone Mineralization, and leaky gut, as well as its role on influencing serum concentrations of peptide YY (PPY) and the ammonia concentration in turkey manure. Two independent trials were conducted. In each experiment, day-of-hatch female turkey poults were neck tagged and randomly assigned to either a control rye-based diet or a rye-based diet supplemented with the DFM (n = 25 birds/group). At 10 days-of-age, poults in both groups were administered with an appropriate dose of fluorescein isothiocyanate-dextran (FITC-d) by oral gavage. One hour later, all poults were euthanized. Blood was collected to evaluate serum FITC-d and PPY concentrations. Furthermore, in Trial 2 only, both tibias were removed for assessment of Bone parameters, and turkey manure was collected to evaluate physicochemical analysis. In both trials, poults treated with the DFM showed a significant increase (P < 0.05) in body weight and body weight gain as compared with control nontreated poults. Poults that received the DFM also had a significant reduction in serum levels of PPY and FITC-d when compared with control nontreated poults. In Trial 2, turkeys treated with the DFM had a substantial increase in tibia strength, tibia diameter, total ash, calcium, and phosphorus when compared with control nontreated turkeys. Their manure was also shown to have a significant reduction in the concentration of ammonia. This is the first report of a commercial DFM reducing the concentration of this compound in turkey manure. In summary, the results of the present study confirm that turkeys fed with a rye-based diet have a significant increase in gut permeability, a reduced body weight, and decreased Bone Mineralization when compared with turkeys fed with the DFM. Turkeys that received the rye-based diet supplemented with the Bacillus-DFM also had a significant reduction in the serum concentration of PPY when compared with control turkeys. This finding suggests a possible prebiotic effect of rye, warranting future studies to test this effect. Further studies to evaluate the microbiota diversity, as well as the concentration of ceca short-chain fatty acids, are also necessary to confirm the reliability of PPY as a potential metabolomic biomarker in poultry.
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rye affects bacterial translocation intestinal viscosity microbiota composition and Bone Mineralization in turkey poults
PLOS ONE, 2015Co-Authors: G Tellez, Juan D Latorre, B M Hargis, V A Kuttappan, Xochitl HernandezvelascoAbstract:Previously, we have reported that rye significantly increased both viscosity and Clostridium perfringens proliferation when compared with corn in an in vitro digestive model. Two independent trials were conducted to evaluate the effect of rye as a source of energy on bacterial translocation, intestinal viscosity, gut microbiota composition, and Bone Mineralization, when compared with corn in turkey poults. In each experiment, day-of-hatch, turkey poults were randomly assigned to either a corn or a rye diet (n = 0 /group). At 10 d of age, in both experiments, 12 birds/group were given an oral gavage dose of fluorescein isothiocyanate dextran (FITC-d). After 2.5 h of oral gavage, blood and liver samples were collected to evaluate the passage of FITC-d and bacterial translocation (BT) respectively. Duodenum, ileum and cecum gut sections were collected to evaluate intestinal viscosity and to enumerate gut microbiota. Tibias were collected for observation of Bone parameters. Broilers fed with a rye diet showed increased (p<0.05) intestinal viscosity, BT, and serum FITC-d. Bacterial enumeration revealed that turkey poults fed with rye had increased the number of total lactic acid bacteria (LAB) in all three sections of the gastrointestinal tract evaluated when compared to turkey poults fed with corn. Turkey poults fed with rye also had significantly higher coliforms in duodenum and ileum but not in the ceca, whereas the total number of anaerobes increased only in duodenum. A significant reduction in Bone strength and Bone Mineralization was observed in turkey poults fed with rye when compared with corn fed turkey poults. In conclusion, rye evoked mucosal damage in turkey poults that increased intestinal viscosity, increased leakage through the intestinal tract, and altered the microbiota composition and Bone Mineralization. Studies to evaluate dietary inclusion of selected Direct-Fed Microbial (DFM) candidates that produce exogenous enzymes in rye fed turkey poults are currently being evaluated.
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role of a bacillus subtilis direct fed microbial on digesta viscosity bacterial translocation and Bone Mineralization in turkey poults fed with a rye based diet
Frontiers in Veterinary Science, 2014Co-Authors: Juan D Latorre, Xochitl Hernandezvelasco, Michael H Kogut, J L Vicente, Ross Wolfenden, A D Wolfenden, B M Hargis, V A Kuttappan, G TellezAbstract:Rye contains high concentrations of non-starch polysaccharides (NSP), leading to reduced digestibility. Since poultry have little or no endogenous enzymes capable of hydrolyzing these NSP, exogenous carbohydrases as feed additives are used in an attempt to reduce the anti-nutritional effects of these polysaccharides. Previously, an in vitro study conducted in our laboratory showed that inclusion of certain Bacillus direct-fed microbial (DFM) candidates that produce exogenous phytase, lipase, protease, cellulase and xylanase in high NSP diets significantly reduced both digesta viscosity and Clostridium perfringens proliferation. In the present study, rye-based turkey starter diets with or without Bacillus-DFM were administered ad libitum to day-of-hatch turkey poults in two independent experiments. In both experiments, day-of-hatch turkey poults were randomly assigned to either a control diet (CON) or a DFM treated diet (n = 25 birds/group). At ten days–of-age, all turkey poults from experiments 1 and 2 were weighted and 12 turkey poults/group were randomly selected and humanely killed. Liver samples were aseptically collected to evaluate bacterial translocation, and intestinal digesta samples were individually collected to evaluate viscosity. Additionally, in experiment 2 both tibias were removed for assessment of Bone parameters. In both experiments, the TRT group showed a reduction in the total number of coliforms in the liver as well as a reduced digesta viscosity when compared to the CON group (P<0.05). Turkey poults fed the Bacillus-DFM candidate had increased tibia diameter, breaking strength, ash content, calcium content, and phosphorus content when compared with CON turkey poults. In summary, turkey poults fed with a rye-based diet without DFM showed an increase in bacterial translocation and digesta viscosity, accompanied by a reduction in Bone Mineralization; however these adverse effects can be prevented by the inclusion of selected a Bacillus-DFM candida
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utilization of rye as energy source affects bacterial translocation intestinal viscosity microbiota composition and Bone Mineralization in broiler chickens
Frontiers in Genetics, 2014Co-Authors: G Tellez, Juan D Latorre, Xochitl Hernandezvelasco, Michael H Kogut, B M Hargis, V A Kuttappan, Amanda Wolfenden, Walter Bottje, L R Bielke, Olivia B FaulknerAbstract:Two independent trials were conducted to evaluate the utilization of rye as energy source on bacterial translocation, intestinal viscosity, gut integrity, gut microbiota composition, and Bone Mineralization, when compared with a traditional cereal (corn) in broiler chickens. In each experiment, day-of-hatch, broiler chickens were randomly assigned to either a corn or a rye diet (n = 20 chickens/group). At 10d of age, in both experiments, 12 chickens/group were randomly selected, and given an oral gavage dose of fluorescein isothiocyanate dextran (FITC-d). After 2.5 h of oral gavage, blood samples were collected to determine the passage of FITC-d. The liver was collected from each bird to evaluate bacterial translocation (BT). Duodenum, ileum and cecum gut sections were collected to evaluate intestinal viscosity and to enumerate gut microbiota. Tibias were collected for observation of Bone parameters. Broilers fed with rye showed increased (p < 0.05) intestinal viscosity, BT, and serum FITC-d. Bacterial enumeration revealed that chickens fed with rye had increased the number of total lactic acid bacteria (LAB) in all three sections of the gastrointestinal tract evaluated when compared to chickens fed with corn. Chickens fed with rye also had significantly higher coliforms in duodenum and ileum, whereas the total number of anaerobes increased only in duodenum. A significant reduction in Bone strength and Bone Mineralization was observed in chickens fed with rye when compared with corn fed chickens. In conclusion, rye evoked mucosal damage in chickens that alter the intestinal viscosity, increased leakage through the intestinal tract, and altered the microbiota composition as well as Bone Mineralization. Studies to evaluate dietary inclusion of selected DFM candidates that produce exogenous enzymes in rye fed chickens are currently being evaluated.
Juan D Latorre - One of the best experts on this subject based on the ideXlab platform.
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effect of bacillus direct fed microbial on leaky gut serum peptide yy concentration Bone Mineralization and ammonia excretion in neonatal female turkey poults fed with a rye based diet
Poultry Science, 2020Co-Authors: G Tellez, Juan D Latorre, Xochitl Hernandezvelasco, B M Hargis, Ruben Merinoguzman, Margarita A Arreguinnava, J A Maguey, M A Michel, P A Moore, Guillermo TellezisaiasAbstract:Rye is high in nonstarch polysaccharides (NSP), a complex carbohydrate which cannot be digested by poultry as they lack the endogenous enzymes to do so. Exogenous carbohydrases must therefore be supplemented to avoid the antinutritional effects associated with a high NSP diet. The objectives of the present study were to evaluate the effects of a rye-based diet with and without supplementation of a Bacillus direct-fed microbial (DFM) on body weight, Bone Mineralization, and leaky gut, as well as its role on influencing serum concentrations of peptide YY (PPY) and the ammonia concentration in turkey manure. Two independent trials were conducted. In each experiment, day-of-hatch female turkey poults were neck tagged and randomly assigned to either a control rye-based diet or a rye-based diet supplemented with the DFM (n = 25 birds/group). At 10 days-of-age, poults in both groups were administered with an appropriate dose of fluorescein isothiocyanate-dextran (FITC-d) by oral gavage. One hour later, all poults were euthanized. Blood was collected to evaluate serum FITC-d and PPY concentrations. Furthermore, in Trial 2 only, both tibias were removed for assessment of Bone parameters, and turkey manure was collected to evaluate physicochemical analysis. In both trials, poults treated with the DFM showed a significant increase (P < 0.05) in body weight and body weight gain as compared with control nontreated poults. Poults that received the DFM also had a significant reduction in serum levels of PPY and FITC-d when compared with control nontreated poults. In Trial 2, turkeys treated with the DFM had a substantial increase in tibia strength, tibia diameter, total ash, calcium, and phosphorus when compared with control nontreated turkeys. Their manure was also shown to have a significant reduction in the concentration of ammonia. This is the first report of a commercial DFM reducing the concentration of this compound in turkey manure. In summary, the results of the present study confirm that turkeys fed with a rye-based diet have a significant increase in gut permeability, a reduced body weight, and decreased Bone Mineralization when compared with turkeys fed with the DFM. Turkeys that received the rye-based diet supplemented with the Bacillus-DFM also had a significant reduction in the serum concentration of PPY when compared with control turkeys. This finding suggests a possible prebiotic effect of rye, warranting future studies to test this effect. Further studies to evaluate the microbiota diversity, as well as the concentration of ceca short-chain fatty acids, are also necessary to confirm the reliability of PPY as a potential metabolomic biomarker in poultry.
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Evaluation of Bone Marrow Adipose Tissue and Bone Mineralization on Broiler Chickens Affected by Wooden Breast Myopathy
Frontiers Media S.A., 2019Co-Authors: Barbara De Almeida Mallmann, Juan D Latorre, Mikayla F A Baxter, Elizabeth M. Martin, Kyung Soo Kim, Norma L. Calderon-apodaca, Xochitl Hernandez-velasco, Leopoldo Paasch-martinez, Casey M. Owens, Sami DridiAbstract:In humans, alterations in Bone metabolism have been associated with myopathies. We postulate the hypothesis that perhaps similar pathologies can also be associated in modern chickens. Hence, this study aimed to assess the fat infiltration in Bone marrow and its repercussion on broiler chicken affected by Wooden Breast (WB) myopathy. Ten Cobb 500 live birds with extreme rigidity of the Pectoralis major (PM) muscle were selected as WB affected chickens by physical examination of the muscle at 49 days of age, whereas ten chickens healthy with no physical signs of hardness in the breast muscle were considered to be unaffected. Macroscopic lesions in affected chickens included areas of firm and inflamed muscle with pale appearance, hemorrhaging, and viscous exudate on the surface. Bone marrow and sections of the PM muscle were collected and analyzed for light microscopy. Additionally, transmission electron microscopy was conducted in affected or unaffected muscle. Chickens affected with WB showed significant reductions (P < 0.05) in femur diameter, calcium, and phosphorous percentage but increased breast weight, compression force and filet thickness when compared with non-affected chickens. Interestingly, Bone marrow from WB chicken had subjectively, more abundant infiltration of adipose tissue, when compared with non-affected chickens. Histology of the Pectoralis major of birds with WB showed abundant infiltration of adipose tissue, muscle fibers degeneration with necrosis and infiltration of heterophils and mononuclear cells, connective tissue proliferation, and vasculitis. Ultrastructural changes of WB muscle revealed lack definition of bands in muscle tissue, or any normal ultrastructural anatomy such as myofibrils. The endomysium components were necrotic, and in some areas, the endomysium was notable only as a string of necrotic tissue between degraded myofibrils. The fascia appeared hypertrophied, with large areas of necrosis and myofiber without structural identity with degraded mitochondria adjacent to the disrupted muscle tissue. As far as we know, this is the first study that describes a subjective increase in adipose tissue in the Bone marrow of chickens affected with WB when compared with non-affected chickens, and reduced Bone Mineralization
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assessment of a nutritional rehabilitation model in two modern broilers and their jungle fowl ancestor a model for better understanding childhood undernutrition
Frontiers in Nutrition, 2018Co-Authors: Mikayla F A Baxter, Juan D Latorre, Sami Dridi, Dawn A Koltes, Elizabeth S Greene, Stephen W Bickler, Jae H Kim, Ruben Merinoguzman, Xochitl HernandezvelascoAbstract:This article is the first in a series of manuscripts to evaluate nutritional rehabilitation in chickens as a model to study interventions in children malnutrition (Part 1: Performance, Bone Mineralization, and Intestinal Morphometric Analysis). Inclusion of rye in poultry diets induces a nutritional deficit that leads to increased bacterial translocation, intestinal viscosity, and decreased Bone Mineralization. However, it is unclear the effect of diet on developmental stage or genetic strain. Therefore, the objective was to determine the effects of a rye diet during either the early or late phase of development on performance, Bone Mineralization, and intestinal morphology across three diverse genetic backgrounds. Modern 2015 (Cobb 500) broiler chicken, 1995 Cobb broiler chicken, and the Giant Jungle Fowl were randomly allocated into four different dietary treatments. Dietary treatments were (1) a control corn-based diet throughout the trial (corn-corn); (2) an early phase malnutrition diet where chicks received a rye-based diet for 10 days, and then switched to the control diet (rye-corn); (3) a malnutrition rye-diet that was fed throughout the trial (rye-rye); and (4) a late phase malnutrition diet where chicks received the control diet for 10 days, and then switched to the rye diet for the last phase (corn-rye). At 10 days of age, chicks were weighed and diets were switched in groups 2 and 4. At day 20 of age, all chickens were weighed and euthanized to collect Bone and intestinal samples. Body weight, weight gain, and Bone Mineralization were different across diet, genetic line, age and all two- and three-way interactions (P < 0.05). Overall, Jungle Fowl were the most tolerant to a rye-based diet, and both the modern and 1995 broilers were significantly affected by the high rye-based diet. However, the 1995 broilers consuming the rye-based diet appeared to experience more permanent effects when compared with the modern broiler. The results of this study suggest that chickens have a great potential as a nutritional rehabilitation model in human trials. The 1995 broilers line was an intermediate genetic line between the fast growing modern line and the non-selected Jungle Fowl line, suggesting that it would be the most appropriate model to study for future studies.
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rye affects bacterial translocation intestinal viscosity microbiota composition and Bone Mineralization in turkey poults
PLOS ONE, 2015Co-Authors: G Tellez, Juan D Latorre, B M Hargis, V A Kuttappan, Xochitl HernandezvelascoAbstract:Previously, we have reported that rye significantly increased both viscosity and Clostridium perfringens proliferation when compared with corn in an in vitro digestive model. Two independent trials were conducted to evaluate the effect of rye as a source of energy on bacterial translocation, intestinal viscosity, gut microbiota composition, and Bone Mineralization, when compared with corn in turkey poults. In each experiment, day-of-hatch, turkey poults were randomly assigned to either a corn or a rye diet (n = 0 /group). At 10 d of age, in both experiments, 12 birds/group were given an oral gavage dose of fluorescein isothiocyanate dextran (FITC-d). After 2.5 h of oral gavage, blood and liver samples were collected to evaluate the passage of FITC-d and bacterial translocation (BT) respectively. Duodenum, ileum and cecum gut sections were collected to evaluate intestinal viscosity and to enumerate gut microbiota. Tibias were collected for observation of Bone parameters. Broilers fed with a rye diet showed increased (p<0.05) intestinal viscosity, BT, and serum FITC-d. Bacterial enumeration revealed that turkey poults fed with rye had increased the number of total lactic acid bacteria (LAB) in all three sections of the gastrointestinal tract evaluated when compared to turkey poults fed with corn. Turkey poults fed with rye also had significantly higher coliforms in duodenum and ileum but not in the ceca, whereas the total number of anaerobes increased only in duodenum. A significant reduction in Bone strength and Bone Mineralization was observed in turkey poults fed with rye when compared with corn fed turkey poults. In conclusion, rye evoked mucosal damage in turkey poults that increased intestinal viscosity, increased leakage through the intestinal tract, and altered the microbiota composition and Bone Mineralization. Studies to evaluate dietary inclusion of selected Direct-Fed Microbial (DFM) candidates that produce exogenous enzymes in rye fed turkey poults are currently being evaluated.
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role of a bacillus subtilis direct fed microbial on digesta viscosity bacterial translocation and Bone Mineralization in turkey poults fed with a rye based diet
Frontiers in Veterinary Science, 2014Co-Authors: Juan D Latorre, Xochitl Hernandezvelasco, Michael H Kogut, J L Vicente, Ross Wolfenden, A D Wolfenden, B M Hargis, V A Kuttappan, G TellezAbstract:Rye contains high concentrations of non-starch polysaccharides (NSP), leading to reduced digestibility. Since poultry have little or no endogenous enzymes capable of hydrolyzing these NSP, exogenous carbohydrases as feed additives are used in an attempt to reduce the anti-nutritional effects of these polysaccharides. Previously, an in vitro study conducted in our laboratory showed that inclusion of certain Bacillus direct-fed microbial (DFM) candidates that produce exogenous phytase, lipase, protease, cellulase and xylanase in high NSP diets significantly reduced both digesta viscosity and Clostridium perfringens proliferation. In the present study, rye-based turkey starter diets with or without Bacillus-DFM were administered ad libitum to day-of-hatch turkey poults in two independent experiments. In both experiments, day-of-hatch turkey poults were randomly assigned to either a control diet (CON) or a DFM treated diet (n = 25 birds/group). At ten days–of-age, all turkey poults from experiments 1 and 2 were weighted and 12 turkey poults/group were randomly selected and humanely killed. Liver samples were aseptically collected to evaluate bacterial translocation, and intestinal digesta samples were individually collected to evaluate viscosity. Additionally, in experiment 2 both tibias were removed for assessment of Bone parameters. In both experiments, the TRT group showed a reduction in the total number of coliforms in the liver as well as a reduced digesta viscosity when compared to the CON group (P<0.05). Turkey poults fed the Bacillus-DFM candidate had increased tibia diameter, breaking strength, ash content, calcium content, and phosphorus content when compared with CON turkey poults. In summary, turkey poults fed with a rye-based diet without DFM showed an increase in bacterial translocation and digesta viscosity, accompanied by a reduction in Bone Mineralization; however these adverse effects can be prevented by the inclusion of selected a Bacillus-DFM candida
Paul Roschger - One of the best experts on this subject based on the ideXlab platform.
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increased fgf 23 levels are linked to ineffective erythropoiesis and impaired Bone Mineralization in myelodysplastic syndromes
JCI insight, 2020Co-Authors: Heike Weidner, Ulrike Baschant, Franziska Lademann, Maria Ledesma G Colunga, Ekaterina Balaian, Christine Hofbauer, Barbara M Misof, Paul Roschger, Stephane BlouinAbstract:Myelodysplastic syndromes (MDS) are clonal malignant hematopoietic disorders in the elderly characterized by ineffective hematopoiesis. This is accompanied by an altered Bone microenvironment, which contributes to MDS progression and higher Bone fragility. The underlying mechanisms remain largely unexplored. Here, we show that myelodysplastic NUP98‑HOXD13 (NHD13) transgenic mice display an abnormally high number of osteoblasts, yet a higher fraction of nonmineralized Bone, indicating delayed Bone Mineralization. This was accompanied by high fibroblast growth factor-23 (FGF-23) serum levels, a phosphaturic hormone that inhibits Bone Mineralization and erythropoiesis. While Fgf23 mRNA expression was low in Bone, brain, and kidney of NHD13 mice, its expression was increased in erythroid precursors. Coculturing these precursors with WT osteoblasts induced osteoblast marker gene expression, which was inhibited by blocking FGF-23. Finally, antibody-based neutralization of FGF-23 in myelodysplastic NHD13 mice improved Bone Mineralization and Bone microarchitecture, and it ameliorated anemia. Importantly, higher serum levels of FGF‑23 and an elevated amount of nonmineralized Bone in patients with MDS validated the findings. C‑terminal FGF‑23 correlated negatively with hemoglobin levels and positively with the amount of nonmineralized Bone. Thus, our study identifies FGF-23 as a link between altered Bone structure and ineffective erythropoiesis in MDS with the prospects of a targeted therapeutic intervention.
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differential effects of teriparatide and zoledronic acid on Bone Mineralization density distribution at 6 and 24 months in the shotz study
Journal of Bone and Mineral Research, 2016Co-Authors: David W Dempster, Barbara M Misof, Paul Roschger, Klaus Klaushofer, Eleftherios P Paschalis, Hua Zhou, Jahangir Alam, Valerie A Ruff, Kathleen A TaylorAbstract:The Skeletal Histomorphometry in Patients on Teriparatide or Zoledronic Acid Therapy (SHOTZ) study assessed the progressive effects of teriparatide (TPTD) and zoledronic acid (ZOL) on Bone remodeling and material properties in postmenopausal women with osteoporosis. Previously, we reported that biochemical and histomorphometric Bone formation indices were significantly higher in patients receiving TPTD versus ZOL. Here we report Bone Mineralization density distribution (BMDD) results based on quantitative backscattered electron imaging (qBEI). The 12-month primary study was randomized and double blind until the month 6 biopsy, then open label. Patients (TPTD, n = 28; ZOL, n = 31) were then eligible to enter a 12-month open-label extension with their original treatment: TPTD 20 μg/d (subcutaneous injection) or ZOL 5 mg/yr (intravenous infusion). A second biopsy was collected from the contralateral side at month 24 (TPTD, n = 10; ZOL, n = 10). In cancellous Bone, ZOL treatment was associated at 6 and 24 months with significantly higher average degree of Mineralization (CaMEAN, +2.2%, p = 0.018; +3.9%, p = 0.009, respectively) and with lower percentage of low mineralized areas (CaLOW , -34.6%, p = 0.029; -33.7%, p = 0.025, respectively) and heterogeneity of Mineralization CaWIDTH (-12.3%, p = 0.003; -9.9%, p = 0.012, respectively), indicating higher Mineralization density and more homogeneous mineral content versus TPTD. Within the ZOL group, significant changes were found in all parameters from month 6 to 24, indicating a progressive increase in Mineralization density. In sharp contrast, Mineralization density did not increase over time with TPTD, reflecting ongoing deposition of new Bone. Similar results were observed in cortical Bone. In this study, TPTD stimulated new Bone formation, producing a mineralized Bone matrix that remained relatively heterogeneous with a stable mean mineral content. ZOL slowed Bone turnover and prolonged secondary Mineralization, producing a progressively more homogeneous and highly mineralized Bone matrix. Although both TPTD and ZOL increase clinical measures of Bone mineral density (BMD), this study shows that the underlying mechanisms of the BMD increases are fundamentally different. © 2016 American Society for Bone and Mineral Research.
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differential effects of teriparatide and zoledronic acid on Bone Mineralization density distribution at 6 and 24 months in the shotz study
Journal of Bone and Mineral Research, 2016Co-Authors: David W Dempster, Barbara M Misof, Paul Roschger, Klaus Klaushofer, Eleftherios P Paschalis, Hua Zhou, Jahangir Alam, Valerie A Ruff, Kathleen A TaylorAbstract:The Skeletal Histomorphometry in Patients on Teriparatide or Zoledronic Acid Therapy (SHOTZ) study assessed the progressive effects of teriparatide (TPTD) and zoledronic acid (ZOL) on Bone remodeling and material properties in postmenopausal women with osteoporosis. Previously, we reported that biochemical and histomorphometric Bone formation indices were significantly higher in patients receiving TPTD versus ZOL. Here we report Bone Mineralization density distribution (BMDD) results based on quantitative backscattered electron imaging (qBEI). The 12-month primary study was randomized and double blind until the month 6 biopsy, then open label. Patients (TPTD, n = 28; ZOL, n = 31) were then eligible to enter a 12-month open-label extension with their original treatment: TPTD 20 μg/d (subcutaneous injection) or ZOL 5 mg/yr (intravenous infusion). A second biopsy was collected from the contralateral side at month 24 (TPTD, n = 10; ZOL, n = 10). In cancellous Bone, ZOL treatment was associated at 6 and 24 months with significantly higher average degree of Mineralization (CaMEAN, +2.2%, p = 0.018; +3.9%, p = 0.009, respectively) and with lower percentage of low mineralized areas (CaLOW, –34.6%, p = 0.029; –33.7%, p = 0.025, respectively) and heterogeneity of Mineralization CaWIDTH (–12.3%, p = 0.003; –9.9%, p = 0.012, respectively), indicating higher Mineralization density and more homogeneous mineral content versus TPTD. Within the ZOL group, significant changes were found in all parameters from month 6 to 24, indicating a progressive increase in Mineralization density. In sharp contrast, Mineralization density did not increase over time with TPTD, reflecting ongoing deposition of new Bone. Similar results were observed in cortical Bone. In this study, TPTD stimulated new Bone formation, producing a mineralized Bone matrix that remained relatively heterogeneous with a stable mean mineral content. ZOL slowed Bone turnover and prolonged secondary Mineralization, producing a progressively more homogeneous and highly mineralized Bone matrix. Although both TPTD and ZOL increase clinical measures of Bone mineral density (BMD), this study shows that the underlying mechanisms of the BMD increases are fundamentally different. © 2016 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals, Inc. on behalf of American Society for Bone and Mineral Research (ASBMR).
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relationship of Bone Mineralization density distribution bmdd in cortical and cancellous Bone within the iliac crest of healthy premenopausal women
Calcified Tissue International, 2014Co-Authors: Barbara M Misof, Paul Roschger, Nadja Fratzlzelman, Peter Fratzl, David W Dempster, Hua Zhou, Shonni J Silverberg, Elizabeth Shane, Adi Cohen, Emily M SteinAbstract:Bone Mineralization density distribution (BMDD) is an important determinant of Bone mechanical properties. The most available skeletal site for access to the BMDD is the iliac crest. Compared to cancellous Bone much less information on BMDD is available for cortical Bone. Hence, we analyzed complete transiliac crest Bone biopsy samples from premenopausal women (n = 73) aged 25–48 years, clinically classified as healthy, by quantitative backscattered electron imaging for cortical (Ct.) and cancellous (Cn.) BMDD. The Ct.BMDD was characterized by the arithmetic mean of the BMDD of the cortical plates. We found correlations between Ct. and Cn. BMDD variables with correlation coefficients r between 0.42 and 0.73 (all p 1wt % between both cortices suggesting a possible modeling situation. This normative dataset of healthy premenopausal women will provide a reference standard by which disease- and treatment-specific effects can be assessed at the level of cortical Bone BMDD.
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Effects of Intermittent Parathyroid Hormone Administration on Bone Mineralization Density in Iliac Crest Biopsies from Patients with Osteoporosis: A Paired Study before and after Treatment
2013Co-Authors: Barbara M Misof, Paul Roschger, David W Dempster, Elizabeth Shane, Felicia Cosman, Etah S. Kurland, Walter Tesch, Phaedra Messmer, Jeri Nieves, Peter FratzlAbstract:Anabolic effects of PTH have been observed at several skeletal sites in humans by dual x-ray absorptiometry without differentiating between an actual increase in Bone volume and an increase in mineral content within already established Bone. The present study addressed this issue by evaluating the Bone Mineralization density distribution of iliac crest Bone biopsies before and after PTH treatment for 18–36 months in men and women with osteoporosis using quantitative backscattered electron imaging. In cortical Bone, pairwise comparison of the two biopsies before and after treatment revealed a reduction in the typical calcium concentration in men (�3.32%; P � 0.02, by paired t test), but no change in women, and the ALBRIGHT (1) ORIGINALLY demonstrated that parathyroid extract was anabolic to the skeletons of rodents. These findings were confirmed in humans in a serie
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effect of bacillus direct fed microbial on leaky gut serum peptide yy concentration Bone Mineralization and ammonia excretion in neonatal female turkey poults fed with a rye based diet
Poultry Science, 2020Co-Authors: G Tellez, Juan D Latorre, Xochitl Hernandezvelasco, B M Hargis, Ruben Merinoguzman, Margarita A Arreguinnava, J A Maguey, M A Michel, P A Moore, Guillermo TellezisaiasAbstract:Rye is high in nonstarch polysaccharides (NSP), a complex carbohydrate which cannot be digested by poultry as they lack the endogenous enzymes to do so. Exogenous carbohydrases must therefore be supplemented to avoid the antinutritional effects associated with a high NSP diet. The objectives of the present study were to evaluate the effects of a rye-based diet with and without supplementation of a Bacillus direct-fed microbial (DFM) on body weight, Bone Mineralization, and leaky gut, as well as its role on influencing serum concentrations of peptide YY (PPY) and the ammonia concentration in turkey manure. Two independent trials were conducted. In each experiment, day-of-hatch female turkey poults were neck tagged and randomly assigned to either a control rye-based diet or a rye-based diet supplemented with the DFM (n = 25 birds/group). At 10 days-of-age, poults in both groups were administered with an appropriate dose of fluorescein isothiocyanate-dextran (FITC-d) by oral gavage. One hour later, all poults were euthanized. Blood was collected to evaluate serum FITC-d and PPY concentrations. Furthermore, in Trial 2 only, both tibias were removed for assessment of Bone parameters, and turkey manure was collected to evaluate physicochemical analysis. In both trials, poults treated with the DFM showed a significant increase (P < 0.05) in body weight and body weight gain as compared with control nontreated poults. Poults that received the DFM also had a significant reduction in serum levels of PPY and FITC-d when compared with control nontreated poults. In Trial 2, turkeys treated with the DFM had a substantial increase in tibia strength, tibia diameter, total ash, calcium, and phosphorus when compared with control nontreated turkeys. Their manure was also shown to have a significant reduction in the concentration of ammonia. This is the first report of a commercial DFM reducing the concentration of this compound in turkey manure. In summary, the results of the present study confirm that turkeys fed with a rye-based diet have a significant increase in gut permeability, a reduced body weight, and decreased Bone Mineralization when compared with turkeys fed with the DFM. Turkeys that received the rye-based diet supplemented with the Bacillus-DFM also had a significant reduction in the serum concentration of PPY when compared with control turkeys. This finding suggests a possible prebiotic effect of rye, warranting future studies to test this effect. Further studies to evaluate the microbiota diversity, as well as the concentration of ceca short-chain fatty acids, are also necessary to confirm the reliability of PPY as a potential metabolomic biomarker in poultry.
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rye affects bacterial translocation intestinal viscosity microbiota composition and Bone Mineralization in turkey poults
PLOS ONE, 2015Co-Authors: G Tellez, Juan D Latorre, B M Hargis, V A Kuttappan, Xochitl HernandezvelascoAbstract:Previously, we have reported that rye significantly increased both viscosity and Clostridium perfringens proliferation when compared with corn in an in vitro digestive model. Two independent trials were conducted to evaluate the effect of rye as a source of energy on bacterial translocation, intestinal viscosity, gut microbiota composition, and Bone Mineralization, when compared with corn in turkey poults. In each experiment, day-of-hatch, turkey poults were randomly assigned to either a corn or a rye diet (n = 0 /group). At 10 d of age, in both experiments, 12 birds/group were given an oral gavage dose of fluorescein isothiocyanate dextran (FITC-d). After 2.5 h of oral gavage, blood and liver samples were collected to evaluate the passage of FITC-d and bacterial translocation (BT) respectively. Duodenum, ileum and cecum gut sections were collected to evaluate intestinal viscosity and to enumerate gut microbiota. Tibias were collected for observation of Bone parameters. Broilers fed with a rye diet showed increased (p<0.05) intestinal viscosity, BT, and serum FITC-d. Bacterial enumeration revealed that turkey poults fed with rye had increased the number of total lactic acid bacteria (LAB) in all three sections of the gastrointestinal tract evaluated when compared to turkey poults fed with corn. Turkey poults fed with rye also had significantly higher coliforms in duodenum and ileum but not in the ceca, whereas the total number of anaerobes increased only in duodenum. A significant reduction in Bone strength and Bone Mineralization was observed in turkey poults fed with rye when compared with corn fed turkey poults. In conclusion, rye evoked mucosal damage in turkey poults that increased intestinal viscosity, increased leakage through the intestinal tract, and altered the microbiota composition and Bone Mineralization. Studies to evaluate dietary inclusion of selected Direct-Fed Microbial (DFM) candidates that produce exogenous enzymes in rye fed turkey poults are currently being evaluated.
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role of a bacillus subtilis direct fed microbial on digesta viscosity bacterial translocation and Bone Mineralization in turkey poults fed with a rye based diet
Frontiers in Veterinary Science, 2014Co-Authors: Juan D Latorre, Xochitl Hernandezvelasco, Michael H Kogut, J L Vicente, Ross Wolfenden, A D Wolfenden, B M Hargis, V A Kuttappan, G TellezAbstract:Rye contains high concentrations of non-starch polysaccharides (NSP), leading to reduced digestibility. Since poultry have little or no endogenous enzymes capable of hydrolyzing these NSP, exogenous carbohydrases as feed additives are used in an attempt to reduce the anti-nutritional effects of these polysaccharides. Previously, an in vitro study conducted in our laboratory showed that inclusion of certain Bacillus direct-fed microbial (DFM) candidates that produce exogenous phytase, lipase, protease, cellulase and xylanase in high NSP diets significantly reduced both digesta viscosity and Clostridium perfringens proliferation. In the present study, rye-based turkey starter diets with or without Bacillus-DFM were administered ad libitum to day-of-hatch turkey poults in two independent experiments. In both experiments, day-of-hatch turkey poults were randomly assigned to either a control diet (CON) or a DFM treated diet (n = 25 birds/group). At ten days–of-age, all turkey poults from experiments 1 and 2 were weighted and 12 turkey poults/group were randomly selected and humanely killed. Liver samples were aseptically collected to evaluate bacterial translocation, and intestinal digesta samples were individually collected to evaluate viscosity. Additionally, in experiment 2 both tibias were removed for assessment of Bone parameters. In both experiments, the TRT group showed a reduction in the total number of coliforms in the liver as well as a reduced digesta viscosity when compared to the CON group (P<0.05). Turkey poults fed the Bacillus-DFM candidate had increased tibia diameter, breaking strength, ash content, calcium content, and phosphorus content when compared with CON turkey poults. In summary, turkey poults fed with a rye-based diet without DFM showed an increase in bacterial translocation and digesta viscosity, accompanied by a reduction in Bone Mineralization; however these adverse effects can be prevented by the inclusion of selected a Bacillus-DFM candida
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utilization of rye as energy source affects bacterial translocation intestinal viscosity microbiota composition and Bone Mineralization in broiler chickens
Frontiers in Genetics, 2014Co-Authors: G Tellez, Juan D Latorre, Xochitl Hernandezvelasco, Michael H Kogut, B M Hargis, V A Kuttappan, Amanda Wolfenden, Walter Bottje, L R Bielke, Olivia B FaulknerAbstract:Two independent trials were conducted to evaluate the utilization of rye as energy source on bacterial translocation, intestinal viscosity, gut integrity, gut microbiota composition, and Bone Mineralization, when compared with a traditional cereal (corn) in broiler chickens. In each experiment, day-of-hatch, broiler chickens were randomly assigned to either a corn or a rye diet (n = 20 chickens/group). At 10d of age, in both experiments, 12 chickens/group were randomly selected, and given an oral gavage dose of fluorescein isothiocyanate dextran (FITC-d). After 2.5 h of oral gavage, blood samples were collected to determine the passage of FITC-d. The liver was collected from each bird to evaluate bacterial translocation (BT). Duodenum, ileum and cecum gut sections were collected to evaluate intestinal viscosity and to enumerate gut microbiota. Tibias were collected for observation of Bone parameters. Broilers fed with rye showed increased (p < 0.05) intestinal viscosity, BT, and serum FITC-d. Bacterial enumeration revealed that chickens fed with rye had increased the number of total lactic acid bacteria (LAB) in all three sections of the gastrointestinal tract evaluated when compared to chickens fed with corn. Chickens fed with rye also had significantly higher coliforms in duodenum and ileum, whereas the total number of anaerobes increased only in duodenum. A significant reduction in Bone strength and Bone Mineralization was observed in chickens fed with rye when compared with corn fed chickens. In conclusion, rye evoked mucosal damage in chickens that alter the intestinal viscosity, increased leakage through the intestinal tract, and altered the microbiota composition as well as Bone Mineralization. Studies to evaluate dietary inclusion of selected DFM candidates that produce exogenous enzymes in rye fed chickens are currently being evaluated.