Evaluation of sample extracting methods of FCSM by Lactobacillus acidophilus based on a UPLC-Q-TOF-MS global metabolomics analysis

Abstract The study of metabolomics requires extracting as many metabolites as possible from a biological sample. This study aimed to determine the optimal method for the extraction of metabolites from solid-state fermented cottonseed meal (FCSM). The UPLC-Q-TOF-MS global metabolomics technology was used to detect the metabolites in FCSM, and the extraction quantity and extraction efficiency of seven different extraction methods, specifically the WA, 50MeOH, 50MeOHB, 50MeCNB, 80MeOHB, 80MeOH and AMF methods were evaluated. The results showed that the number of VIP metabolites extracted by AMF method are 196 and 184 in ESI+ and ESI- mode respectively, it is the largest number of all exacted methods; and the AMF methods also provided a higher extraction efficiency compared with the other methods, especially in indoleacrylic acid, dl-tryptophan and epicatechin (p 0.01). As a result, AMF/-4 °C method was identified as the best method for the extraction of metabolites from FCSM by Lactobacillus acidophilus. Our study establishes a technical basis for future metabolomics research of fermented feed.

Evaluation of sample extracting methods of FCSM by Lactobacillus acidophilus based on a UPLC-Q-TOF-MS global metabolomics analysis

The study of metabolomics requires extracting as many metabolites as possible from a biological sample. This study aimed to determine the optimal method for the extraction of metabolites from solid-state fermented cottonseed meal (FCSM). The UPLC-Q-TOF-MS global metabolomics technology was used to detect the metabolites in FCSM, and the extraction quantity and extraction efficiency of seven different extraction methods, specifically the WA, 50MeOH, 50MeOHB, 50MeCNB, 80MeOHB, 80MeOH and AMF methods were evaluated. The results showed that the number of VIP metabolites extracted by AMF method are 196 and 184 in ESI+ and ESI- mode respectively, it is the largest number of all exacted methods; and the AMF methods also provided a higher extraction efficiency compared with the other methods, especially in indoleacrylic acid, dl-tryptophan and epicatechin (p < 0.01). As a result, AMF/-4 °C method was identified as the best method for the extraction of metabolites from FCSM by Lactobacillus acidophilus. Our study establishes a technical basis for future metabolomics research of fermented feed.(AU)

Effect of Cottonseed Meal Fermented with Yeast on the Lipid-related Gene Expression in Broiler Chickens

ABSTRACT Fermented cottonseed meal (FCSM) is widely used in poultry diets in China. This study was conducted to investigate the effect of FCSM on lipid-related gene expression in broilers. Initially, 180 broiler chickens (21-days-old, equal number of males and females) were randomly divided into three groups, with six pens per group and 10 birds per pen. The chickens in the control group were fed a diet containing unfermented cottonseed meal, and those in the treatment groups were fed with diets including either CSM fermented by Candida tropicalis (Ct group) or CSM fermented by Candida tropicalis plus Saccharomyces cerevisae (Ct-Sc group) until 64 days old. The results revealed that, compared with the control group (p 0.05), the mRNA expression of peroxisome proliferator-activated receptor alpha (PPAR-a) and lipoprotein lipase (LPL) were upregulated in the livers of Ct-Sc males. The expression of PPAR-a was also upregulated in the livers of Ct females. The expression levels of acetyl CoA carboxylase (ACC) and LPL in the liver of males and the expression of PPAR-a in the liver of females were significantly different between the Ct and Ct-Sc groups (p 0.05). However, gene expressions of fatty acid synthase (FAS) and liver fatty acid-binding protein (L-FABP) in the liver were not altered when the broilers were fed FCSM-supplemented diets (p>0.05). Likewise, the expressions of peroxisome proliferator-activated receptor gamma (PPAR-g) and LPL in the abdominal fat were not altered by the FCSM-supplemented diets (p>0.05). The results in this study indicate that CSM fermented by Candida tropicalis and Saccharomyces cerevisiaeeffectively regulated the genes involved in fatty acid b-oxidation and triglyceride hydrolysis in male broiler chickens. Furthermore, the effects of the FCSM-supplemented diets were significantly different between bird sexes and between yeast strains used in the fermentation process.

Effect of Cottonseed Meal Fermented with Yeast on the Lipid-related Gene Expression in Broiler Chickens

ABSTRACT Fermented cottonseed meal (FCSM) is widely used in poultry diets in China. This study was conducted to investigate the effect of FCSM on lipid-related gene expression in broilers. Initially, 180 broiler chickens (21-days-old, equal number of males and females) were randomly divided into three groups, with six pens per group and 10 birds per pen. The chickens in the control group were fed a diet containing unfermented cottonseed meal, and those in the treatment groups were fed with diets including either CSM fermented by Candida tropicalis (Ct group) or CSM fermented by Candida tropicalis plus Saccharomyces cerevisae (Ct-Sc group) until 64 days old. The results revealed that, compared with the control group (p 0.05), the mRNA expression of peroxisome proliferator-activated receptor alpha (PPAR-a) and lipoprotein lipase (LPL) were upregulated in the livers of Ct-Sc males. The expression of PPAR-a was also upregulated in the livers of Ct females. The expression levels of acetyl CoA carboxylase (ACC) and LPL in the liver of males and the expression of PPAR-a in the liver of females were significantly different between the Ct and Ct-Sc groups (p 0.05). However, gene expressions of fatty acid synthase (FAS) and liver fatty acid-binding protein (L-FABP) in the liver were not altered when the broilers were fed FCSM-supplemented diets (p>0.05). Likewise, the expressions of peroxisome proliferator-activated receptor gamma (PPAR-g) and LPL in the abdominal fat were not altered by the FCSM-supplemented diets (p>0.05). The results in this study indicate that CSM fermented by Candida tropicalis and Saccharomyces cerevisiaeeffectively regulated the genes involved in fatty acid b-oxidation and triglyceride hydrolysis in male broiler chickens. Furthermore, the effects of the FCSM-supplemented diets were significantly different between bird sexes and between yeast strains used in the fermentation process.

Effect of Cottonseed Meal Fermented with Yeast on the Lipid-related Gene Expression in Broiler Chickens

Fermented cottonseed meal (FCSM) is widely used in poultry diets in China. This study was conducted to investigate the effect of FCSM on lipid-related gene expression in broilers. Initially, 180 broiler chickens (21-days-old, equal number of males and females) were randomly divided into three groups, with six pens per group and 10 birds per pen. The chickens in the control group were fed a diet containing unfermented cottonseed meal, and those in the treatment groups were fed with diets including either CSM fermented by Candida tropicalis (Ct group) or CSM fermented by Candida tropicalis plus Saccharomyces cerevisae (Ct-Sc group) until 64 days old. The results revealed that, compared with the control group (p 0.05), the mRNA expression of peroxisome proliferator-activated receptor alpha (PPAR-a) and lipoprotein lipase (LPL) were upregulated in the livers of Ct-Sc males. The expression of PPAR-a was also upregulated in the livers of Ct females. The expression levels of acetyl CoA carboxylase (ACC) and LPL in the liver of males and the expression of PPAR-a in the liver of females were significantly different between the Ct and Ct-Sc groups (p 0.05). However, gene expressions of fatty acid synthase (FAS) and liver fatty acid-binding protein (L-FABP) in the liver were not altered when the broilers were fed FCSM-supplemented diets (p>0.05). Likewise, the expressions of peroxisome proliferator-activated receptor gamma (PPAR-g) and LPL in the abdominal fat were not altered by the FCSM-supplemented diets (p>0.05). The results in this study indicate that CSM fermented by Candida tropicalis and Saccharomyces cerevisiaeeffectively regulated the genes involved in fatty acid b-oxidation and triglyceride hydrolysis in male broiler chickens. Furthermore, the effects of the FCSM-supplemented diets were significantly different between bird sexes and between yeast strains used in the fermentation process.

Effect of Cottonseed Meal Fermented with Yeast on the Lipid-related Gene Expression in Broiler Chickens

Fermented cottonseed meal (FCSM) is widely used in poultry diets in China. This study was conducted to investigate the effect of FCSM on lipid-related gene expression in broilers. Initially, 180 broiler chickens (21-days-old, equal number of males and females) were randomly divided into three groups, with six pens per group and 10 birds per pen. The chickens in the control group were fed a diet containing unfermented cottonseed meal, and those in the treatment groups were fed with diets including either CSM fermented by Candida tropicalis (Ct group) or CSM fermented by Candida tropicalis plus Saccharomyces cerevisae (Ct-Sc group) until 64 days old. The results revealed that, compared with the control group (p 0.05), the mRNA expression of peroxisome proliferator-activated receptor alpha (PPAR-a) and lipoprotein lipase (LPL) were upregulated in the livers of Ct-Sc males. The expression of PPAR-a was also upregulated in the livers of Ct females. The expression levels of acetyl CoA carboxylase (ACC) and LPL in the liver of males and the expression of PPAR-a in the liver of females were significantly different between the Ct and Ct-Sc groups (p 0.05). However, gene expressions of fatty acid synthase (FAS) and liver fatty acid-binding protein (L-FABP) in the liver were not altered when the broilers were fed FCSM-supplemented diets (p>0.05). Likewise, the expressions of peroxisome proliferator-activated receptor gamma (PPAR-g) and LPL in the abdominal fat were not altered by the FCSM-supplemented diets (p>0.05). The results in this study indicate that CSM fermented by Candida tropicalis and Saccharomyces cerevisiaeeffectively regulated the genes involved in fatty acid b-oxidation and triglyceride hydrolysis in male broiler chickens. Furthermore, the effects of the FCSM-supplemented diets were significantly different between bird sexes and between yeast strains used in the fermentation process.(AU)

Uso de inóculo de fezes como substituição do conteúdo ruminal de bubalinos na técnica in vitro de produção de gases / Use of faeces as inoculum as alternative for buffalo rumen contents in the in vitro gas production technique

Com o objetivo de estudar o uso de inóculo de fezes em substituição do conteúdo ruminal de bubalinos na técnica in vitro de produção de gases, este presente trabalho comparou as avaliações realizadas em três ensaios. Foram utilizados três bubalinos da raça Mediterrâneo, machos, adultos, castrados, fistulados no rúmen com média de peso vivo de 450 (± 18,7) kg. Estes animais receberam uma dieta basal, composta de silagem de milho (70%) e concentrado (30%). Estes animais foram os doadores dos 2 tipos de inóculos, que foram conteúdo ruminal (CR) e fezes. O primeiro ensaio realizado foi com alimentos concentrados: grão de milho, farelo de soja, farelo de trigo e farelo de algodão; o segundo ensaio foi com leguminosas forrageiras: alfafa (Medicago sativa L. ), estilosantes pioneiro (Stylosanthes macrocephala cv. Pioneiro), soja perene (Neonotonia wightii) e leucena (Leucaena leucocephala); e o terceiro e último ensaio foi realizado com gramíneas forrageiras: capim braquiarão (Brachiaria brizantha cv. Marandu), capim buffel (Cenchrus ciliaris L. cv. Biloela), estrela africana (Cynodon plectostachyus) e capim mombaça (Panicum maximum Jacq. cv. Mombaça). Os valores médios obtidos de produção potencial de gases em cada ensaio foram menores (P<0,05) para as amostras fermentadas com inóculos de fezes que com conteúdo ruminal, sendo respectivamente, para concentrados (140,23 e 194,08 mL.g-1MS), gramíneas (161,99 e 230,25 mL.g-1MS) e leguminosas (141,78 e 170,70 mL.gm-1MS). Conclui-se que inóculos de fezes não apresentam condições satisfatórias para substituição do inóculum com conteúdo ruminal para uso na técnica de produção in vitro de gases.

In order to study the use of faeces as inoculum as alternative for buffalo rumen contents in the in vitro gas production technique, the present work evaluations of three tests. We used three Mediterranean buffalos, male, adult, neutered, fistulated in the rumen, with an average live weight of 450 (± 18.7) kg. These animals received a basal diet composed of corn silage (70%) and concentrated (30%). These buffalo were the donors of the two types of inocula, rumen content (CR) and faeces. The first test was carried out with concentrate foods: corn grain, soybean meal, wheat bran and cottonseed meal, the second test was with legumes: alfalfa (Medicago sativa L. ), Pioneiro estilo (Stylosanthes macrocephala cv. Pioneiro) , perennial soybean (Neonotonia wightii) and leucaena (Leucaena leucocephala) and the third and last test was carried out with grasses: Marandu grass (Brachiaria brizantha cv. Marandu), buffel grass (Cenchrus ciliaris L. cv. Biloela), African Star grass (Cynodon plectostachyus) and Mombasa grass (Panicum maximum Jacq. cv. Mombasa). The average values of potential production of gas in each test were lower (P<0.05) for samples fermented with an inoculum of faeces with rumen contents, being respectively, for concentrates (140.23 and 194.08 mL.g-1MS), grasses (161.99 and 230.25 mL.g-1MS) and legumes (141.78 and 170.70 mL.g-1MS). It follows that fecal inoculum unsatisfactory condition for replacing the inoculum with rumen technique for use in the in vitro production of gases technique.

Uso de inóculo de fezes como substituição do conteúdo ruminal de bubalinos na técnica in vitro de produção de gases / Use of faeces as inoculum as alternative for buffalo rumen contents in the in vitro gas production technique

Com o objetivo de estudar o uso de inóculo de fezes em substituição do conteúdo ruminal de bubalinos na técnica in vitro de produção de gases, este presente trabalho comparou as avaliações realizadas em três ensaios. Foram utilizados três bubalinos da raça Mediterrâneo, machos, adultos, castrados, fistulados no rúmen com média de peso vivo de 450 (± 18,7) kg. Estes animais receberam uma dieta basal, composta de silagem de milho (70%) e concentrado (30%). Estes animais foram os doadores dos 2 tipos de inóculos, que foram conteúdo ruminal (CR) e fezes. O primeiro ensaio realizado foi com alimentos concentrados: grão de milho, farelo de soja, farelo de trigo e farelo de algodão; o segundo ensaio foi com leguminosas forrageiras: alfafa (Medicago sativa L. ), estilosantes pioneiro (Stylosanthes macrocephala cv. Pioneiro), soja perene (Neonotonia wightii) e leucena (Leucaena leucocephala); e o terceiro e último ensaio foi realizado com gramíneas forrageiras: capim braquiarão (Brachiaria brizantha cv. Marandu), capim buffel (Cenchrus ciliaris L. cv. Biloela), estrela africana (Cynodon plectostachyus) e capim mombaça (Panicum maximum Jacq. cv. Mombaça). Os valores médios obtidos de produção potencial de gases em cada ensaio foram menores (P<0,05) para as amostras fermentadas com inóculos de fezes que com conteúdo ruminal, sendo respectivamente, para concentrados (140,23 e 194,08 mL.g-1MS), gramíneas (161,99 e 230,25 mL.g-1MS) e leguminosas (141,78 e 170,70 mL.gm-1MS). Conclui-se que inóculos de fezes não apresentam condições satisfatórias para substituição do inóculum com conteúdo ruminal para uso na técnica de produção in vitro de gases. (AU)

In order to study the use of faeces as inoculum as alternative for buffalo rumen contents in the in vitro gas production technique, the present work evaluations of three tests. We used three Mediterranean buffalos, male, adult, neutered, fistulated in the rumen, with an average live weight of 450 (± 18.7) kg. These animals received a basal diet composed of corn silage (70%) and concentrated (30%). These buffalo were the donors of the two types of inocula, rumen content (CR) and faeces. The first test was carried out with concentrate foods: corn grain, soybean meal, wheat bran and cottonseed meal, the second test was with legumes: alfalfa (Medicago sativa L. ), Pioneiro estilo (Stylosanthes macrocephala cv. Pioneiro) , perennial soybean (Neonotonia wightii) and leucaena (Leucaena leucocephala) and the third and last test was carried out with grasses: Marandu grass (Brachiaria brizantha cv. Marandu), buffel grass (Cenchrus ciliaris L. cv. Biloela), African Star grass (Cynodon plectostachyus) and Mombasa grass (Panicum maximum Jacq. cv. Mombasa). The average values of potential production of gas in each test were lower (P<0.05) for samples fermented with an inoculum of faeces with rumen contents, being respectively, for concentrates (140.23 and 194.08 mL.g-1MS), grasses (161.99 and 230.25 mL.g-1MS) and legumes (141.78 and 170.70 mL.g-1MS). It follows that fecal inoculum unsatisfactory condition for replacing the inoculum with rumen technique for use in the in vitro production of gases technique. (AU)