Cottonseed meal fermented by Candida tropical reduces the fat deposition in white-feather broilers through cecum bacteria-host metabolic cross-talk.

In the present study, effects of cottonseed meal fermented by Candida tropicalis (FCSM) on fat deposition, cecum microbiota, and metabolites and their interactions were studied in broilers. A total of 180 1-day-old broilers were randomly assigned into two groups with six replicates of 15 birds in each. The birds were offered two diets consisted one control, i.e., supplemented with 0% FCSM (CON) and an experimental, with 6% FCSM (FCSM). Illumina MiSeq sequencing and liquid chromatography-mass spectrometry were used to investigate the profile changes of the cecum microbes and metabolites and the interactions among fat deposition, microbes, and metabolites. Results showed that at the age of 21 days, both the abdominal fat and subcutaneous fat thickness of the experimental birds decreased significantly (P < 0.05) in response to the dietary FCSM supplementation. The predominant microbial flora in cecum consisted Bacteroidetes (53.55%), Firmicutes (33.75%), and Proteobacteria (8.61%). FCSM diet increased the relative abundance of Bacteroides but decreased obese microbial including Faecalibacterium, Lachnospiraceae, Ruminococcaceae, and Anaerofilum. Cecum metabolomics analysis revealed that lipids, organic acids, vitamins, and peptides were significantly altered by adding FCSM in diet. Correlation analysis showed that abdominal fat and subcutaneous fat thickness related negatively with Bacteroides while the same related positively with Faecalibacterium, Lachnospiraceae, and Ruminococcaceae. Moreover, abdominal fat and subcutaneous fat thickness were related negatively with nicotinic acid, sebacic acid, thymidine, and succinic acid. These findings indicated that FCSM reduced the fat deposition by regulating cecum microbiota and metabolites in broilers. The results are contributory to the development of probiotics and the improvement in the production of broilers.

Impacts of Dietary Protein from Fermented Cottonseed Meal on Lipid Metabolism and Metabolomic Profiling in the Serum of Broilers.

Dietary protein from fermented cottonseed meal (FCSM), widely used in poultry diets in China, had regulating effects on lipid metabolism. To understand the effects of FCSM on lipid metabolism in broilers, we analyzed the biochemical indexes, enzyme activity, hormone level and metabolites in serum responses to FCSM intake. One hundred and eighty 21-d-old Chinese yellow feathered broilers (536.07±4.43 g) were randomly divided into 3 groups with 6 replicates and 3 diets with 6 % supplementation of unfermented CSM (control group), FCSM by C. Tropicalis (Ct CSM) or C. tropicalis plus S. Cerevisae (Ct-Sc CSM). Result showed that: (1) FCSM intake decreased significantly the content of triglyceride (TAG), total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) (P<0.05) in serum; (2) FCSM intake could significantly increase enzyme activity of acetyl CoA carboxylase (ACC), lipoprotein lipase (LPL), fatty acid synthase (FAS) and hormone sensitive lipase (HSL) (P<0.05); (3) Ct-Sc CSM intake increased significantly the levels of adiponectin (ADP) (P<0.05); (4) FCSM intake caused significant metabolic changes involving glycolysis, TCA cycle, synthesis of fatty acid and glycogen, and metabolism of glycerolipid, vitamins B group and amino acids. Our results strongly suggested that FCSM intake could significantly affect lipid metabolism via multiple pathways. These findings provided new essential information about the effect of FCSM on broilers and demonstrated the great potential of nutrimetabolomics, through which the research complex nutrients are included in animal diet.

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.

Tissue lipid metabolism and hepatic metabolomic profiling in response to supplementation of fermented cottonseed meal in the diets of broiler chickens.

This study investigated the effects of fermented cottonseed meal (FCSM) on lipid metabolites, lipid metabolism-related gene expression in liver tissues and abdominal adipose tissues, and hepatic metabolomic profiling in broiler chickens. One hundred and eighty 21-d-old broiler chickens were randomly divided into three diet groups with six replicates of 10 birds in each group. The three diets consisted of a control diet supplemented with unfermented cottonseed meal, an experimental diet of cottonseed meal fermented by Candida tropicalis, and a second experimental diet of cottonseed meal fermented by C. tropicalis plus Saccharomyces cerevisae. The results showed that FCSM intake significantly decreased the levels of abdominal fat and hepatic triglycerides (P<0.05 for both). Dietary FCSM supplementation down-regulated the mRNA expression of fatty acid synthase and acetyl CoA carboxylase in liver tissues and the lipoprotein lipase expression in abdominal fat tissues (P<0.05 for both). FCSM intake resulted in significant metabolic changes of multiple pathways in the liver involving the tricarboxylic acid cycle, synthesis of fatty acids, and the metabolism of glycerolipid and amino acids. These findings indicated that FCSM regulated lipid metabolism by increasing or decreasing the expression of the lipid-related gene and by altering multiple endogenous metabolites. Lipid metabolism regulation is a complex process, this discovery provided new essential information about the effects of FCSM diets in broiler chickens and demonstrated the great potential of nutrimetabolomics in researching complex nutrients added to animal diets.