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This study was conducted to evaluate the prebiotic effects of dietary xylooligosaccharide (XOS) supplementation on performance, nutrient digestibility, intestinal morphology, and gut microbiota in laying hens. In a 12-wk experiment, a total of 288 Hy-Line Brown layers at 50 wk of age were randomly assigned into 3 dietary treatments supplemented with XOS at 0, 200 or 400 mg/kg. Each treatment had 8 replicates with 12 birds each. Hens fed XOS diets showed a lower feed-to-egg ratio during wk 7 to 12 and a higher egg yolk color value in wk 12 compared with those fed the control diet (P

Developing new sources of organic selenium (Se) has potential benefits for animal production and human nutrition via animal-based foods enriched with Se. The objective of this study was to evaluate the effects of Se-enriched insect protein (SEIP) in comparison with other sources, such as sodium selenite (SS) and selenium-enriched yeast (SEY), on performance, egg quality, selenium concentration in eggs, serum biochemical indices, immune capacity, and intestinal morphology of laying hens. Four hundred and fifty 24-week-old Hy-Line Brown laying hens with 94.0 ± 1.5% laying rate were randomly allocated to five groups with six replicates of 15 hens each. The control diet was prepared without adding exogenous selenium (calculated basal Se content of 0.08 mg/kg). The normal group was fed basal diets supplemented with 0.3 mg/kg of Se provided by sodium selenite. Three treatment groups (SS, SEY, and SEIP, respectively) were fed basal diets supplemented with 2 mg/kg of Se provided by sodium selenite, Se-enriched yeast, and SEIP, respectively. The feeding trial lasted for 12 weeks. Results revealed that dietary supplementation of 2 mg/kg of Se increased egg weight, decreased feed conversion ratio, and enhanced the antioxidant capacity of eggs in laying hens relative to the control group, whereas no significant differences were observed among SS, SEY, and SEIP treatment groups for the same. The organic source of Se provided by SEY or SEIP showed higher bio efficiency, as indicated by higher selenium content in eggs of SEY and SEIP compared with SS, although higher content was observed in SEY compared with SEIP. Also, the organic Se source significantly improved antioxidant capacity and immune functions of laying hens than the inorganic Se source. Diets supplemented with SEIP and SS significantly improved jejunal morphology of the laying hens compared with SEY, whereas SEIP was more effective than SEY to improve the oviduct health of laying hens. The results of this work evidently points the additive effect and nontoxicity of SEIP. Thus, SEIP could be used as another organic source of Se in the diet of laying hens and production of selenium-enriched eggs for humans.

The objective of this study was to evaluate the effect of dietary chloride (Cl) levels on performance, eggshell mechanical quality, and ultrastructure in layers based on the substitution of sodium chloride (NaCl) by sodium sulfate (Na2SO4). Three hundred sixty Jing Brown laying hens aged 43 wk were randomly divided into 5 groups and fed with corn–soybean meal diets containing 0.06, 0.10, 0.15, 0.20, and 0.25% total Cl inclusion. Every group had 8 replicates of 9 birds each. The feeding trial lasted for 12 wk. The results showed dietary 0.06% Cl due to complete substitution of NaCl by Na2SO4, depressed performance (P < 0.05) from 45 to 54 wk of age, increased serum creatinine level (P < 0.05), and caused visible renal tubular atrophy. Dietary Cl levels quadratically affected breaking strength, thickness, and weight of eggshell (P < 0.05). Better eggshell quality could be obtained when NaCl was partly replaced by Na2SO4 in laying hen diets maintaining Cl level at 0.10 or 0.15%. Moreover, the eggshell ash content was affected by Cl levels in a quadratic (P < 0.001) manner, with higher values observed in the 0.10 and 0.15% Cl groups (P < 0.05). Besides, the eggshell ultrastructural data showed that the total thickness and effective thickness significantly increased (P < 0.05) and mammillary thickness decreased (P < 0.05) in the group of dietary 0.15% Cl compared with the groups of 0.06 and 0.25% Cl. In conclusion, the complete substitution of dietary NaCl by Na2SO4 may induce Cl deficiency and depress laying performance and eggshell quality. Na2SO4 could partly replace NaCl in diets for laying hens (43–54 wk of age) without adverse effects on performance at the dietary Cl level from 0.10 to 0.25%. Better eggshell quality could be obtained when NaCl was partly replaced by Na2SO4 in laying hen diets maintaining Cl level at ∼0.15%.

This study was designed to evaluate the effect of dietary Ser on performance, egg quality, serum indices, and ileal mucosal immunity in laying hens fed low crude protein (LCP), essential amino acids (EAA) balanced diets. A total of 480 Hy-Line Brown layers at 24 wk of age were randomly assigned into 5 dietary treatments with 8 replicates of 12 birds each. Treatments included a control diet (16.49% CP), and 4 LCP, EAA balanced diets (14.05% CP) supplemented with 0, 0.114%, 0.306%, 0.498% L-Ser, respectively. Dietary Ser supplementation linearly increased hen-day egg production (HDEP; P < 0.05) and decreased feed-to-egg ratio (P < 0.05) among LCP groups from wk 6 to 10, and the optimal HDEP of layers occurred at Ser level of 0.498%. At the end of wk 10, birds in the control had higher albumen height and thick white proportion than those fed the LCP diet without Ser addition (P < 0.05), and presented a lower yolk color score than all LCP groups (P < 0.05). Among LCP groups, serum total protein and globulin contents were significantly increased by dietary Ser addition at the levels of 0.306% and 0.498% (P < 0.05), and had a linear response to the supplemental Ser levels (P < 0.05). Furthermore, dietary 0.498% Ser supplementation significantly increased serum immunoglobulin G and immunoglobulin M contents (P < 0.05) and up-regulated the expression of mucin 2, secretory immunoglobulin A, and relevant glycosyltransferases of O-glycosylation in ileal mucosa (P < 0.05). The increased expression of proinflammatory cytokines IFN-γ and IL-1β induced by LCP diets (P < 0.05) was reversed following 0.498% Ser addition (P < 0.05). Collectively, dietary CP reduction by 2.44% could maintain the productive performance of layers when it was fortified with certain EAA, though poor albumen quality, and ileal inflammation were occurred. The addition of Ser to LCP diets improved performance probably through enhancing humoral and ileal mucosal immunity and attenuating the ileal inflammation of layers.

This study investigated the effects of dietary chloride (Cl) reduction on laying performance and eggshell quality by substitution of sodium bicarbonate (NaHCO3) or sodium sulfate (Na2SO4) for part of dietary sodium chloride (NaCl), and further explored its mechanism for improving eggshell quality. A total of 360 29-wk-old Hy-line Brown laying hens were randomly allocated to 5 dietary treatments, including a basal diet contained 0.33% NaCl (control group, 0.27% dietary Cl), and 4 experimental diets that contained 0.21% and 0.15% dietary Cl by substituting Na2SO4 or NaHCO3 for part of NaCl in the basal diet. No significant differences were observed in blood Na+, Cl−, K+ and Ca2+ levels and pH value as well as serum creatinine and uric acid contents among 5 treatments (P > 0.05). Dietary Cl reduction increased egg production and ADFI during wk 33 to 36, 37 to 40 and 29 to 40 of age and decreased feed conversion ratio during wk 37 to 40 of age (P < 0.05). The hens fed with diets containing 0.15% Cl increased eggshell breaking strength, thickness and weight ratio in wk 40 of age (P 0.05). Overall, dietary Cl reductions could improve laying performance and eggshell quality by substitution of NaHCO3 or Na2SO4 for part of NaCl, and there were no differences in the improvements between these two substitutes. The improved eggshell quality may be attributed to improved eggshell ultrastructure and increased supply of eggshell Ca2CO3.

The current experiment was conducted to investigate the application effects of selenium conjugated to insect protein (SCIP) in the production of selenium-enriched eggs. A total of 450 laying hens were randomly assigned to five dietary groups, each group consisting of six replicates. Hens in the control group received a diet without selenium supplementation, whereas hens in the other four groups received diets supplemented with either 1, 2, 5, or 10 mg/kg of selenium from SCIP. The productive performance, egg quality, antioxidant and immune capacity, biochemical indices, intestinal morphology, and oviduct health of laying hens were evaluated. The results showed that the supplementation of organic selenium provided by SCIP in the diets of laying hens enhanced performance and egg quality without any toxicity effect, even at the 10 mg/kg inclusion level. A level of 2 mg/kg of selenium provided by SCIP in diets tentatively improved the serum antioxidant and immune capacity, intestinal development, and oviduct health of laying hens in a conspicuous manner. Hence, the biosafety and positive effects of SCIP as a feed additive supplement in laying hens’ diet have been demonstrated with the enhanced production of safe and selenium-enriched eggs.

The objective of this study was to evaluate the effects of dietary Bacillus subtilis supplementation and calcium (Ca) levels on performance, eggshell quality, intestinal morphology, and relative calbindin-D28k (CALB1) mRNA level of laying hens in the late phase of production. An experiment employing a 2 × 3 factorial arrangement of 3 levels of Ca (3.5, 4.0, and 4.5%) and the absence or presence of B. subtilis was carried out with a total of 576 Hy-Line Brown laying hens aged 72 to 79 wk. Every group had 8 replicates of 12 birds each. The results showed that 4.0 and 4.5% Ca levels improved (P < 0.05) apparent retention and serum Ca content of aged laying hens. Compared with the 3.5% Ca level, the 4.0% Ca level in diets increased (P < 0.05) thickness, eggshell weight, shell ratio, and eggshell Ca content of aged laying hens. Moreover, breaking strength, thickness, eggshell weight, shell ratio, eggshell Ca content, apparent retention of Ca in g/day, apparent retention of Ca in percent, villus height, villus height/crypt depth, serum Ca level, and relative CALB1 mRNA level of aged laying hens were all increased (P < 0.05) by B. subtilis supplementation in diets. The supplemental B. subtilis decreased feed conversion ratio (P = 0.001) significantly. In addition, there was an interaction effect between increased Ca levels from 3.5 to 4.5% and B. subtilis supplementation on crypt depth in the duodenum (P < 0.05). In conclusion, we found that both the increase in dietary Ca level from 3.5 to 4.5% and B. subtilis supplementation could enhance intestinal Ca absorption and improve eggshell quality of laying hens in the late phase of production (72-79 wk of age). Dietary supplementation of B. subtilis accompanying the 4.0% Ca level was appropriate in enhancement of eggshell quality.

Our previous study indicated that dietary xylooligosaccharide (XOS) supplementation improved feed efficiency, ileal morphology, and nutrient digestibility in laying hens. The objective of this study was to evaluate the mitigative effects of XOS on intestinal mucosal barrier impairment and microbiota dysbiosis induced by oxidized fish oil (OFO) in laying hens. A total of 384 Hy-Line Brown layers at 50 weeks of age were randomly divided into four dietary treatments, including the diets supplemented with 20 g/kg of fresh fish oil (FFO group) or 20 g/kg of oxidized fish oil (OFO group), and the OFO diets with XOS addition at 200 mg/kg (OFO/XOS200 group) or 400 mg/kg (OFO/XOS400 group). Each treatment had eight replicates with 12 birds each. The OFO treatment decreased (P < 0.05) the production performance of birds from 7 to 12 weeks of the experiment, reduced (P < 0.05) ileal mucosal secretory immunoglobulin A (sIgA) content, and increased (P < 0.05) serum endotoxin concentration, as well as downregulated (P < 0.05) mRNA expression of claudin-1 (CLDN1) and claudin-5 (CLDN5) in the ileal mucosa at the end of the experiment. Dietary XOS addition (400 mg/kg) recovered (P < 0.05) these changes and further improved (P < 0.05) ileal villus height (VH) and the villus height-to-crypt depth ratio (VCR). In addition, OFO treatment altered cecal microbial composition of layers, and these alterations were probably involved in OFO-induced ileal mucosal impairment as causes or consequences. Supplemental XOS remodeled cecal microbiota of layers fed the OFO diet, characterized by an elevation in microbial richness and changes in microbial composition, including increases in Firmicutes, Ruminococcaceae, Verrucomicrobia (Akkermansia), Paraprevotella, Prevotella_9, and Oscillospira, along with a decrease in Erysipelatoclostridium. The increased abundance of Verrucomicrobia (Akkermansia) had positive correlations with the improved ileal VH and ileal mucosal expression of CLDN1. The abundance of Erysipelatoclostridium decreased by XOS addition was negatively associated with ileal VH, VCR, ileal mucosal sIgA content, and the relative expression of zonula occludens-2, CLDN1, and CLDN5. Collectively, supplemental XOS alleviated OFO-induced intestinal mucosal barrier dysfunction and performance impairment in laying hens, which could be at least partially attributed to the modulation of gut microbiota.

BackgroundLower eggshell quality in the late laying period leads to economic loss. It is a major threat to the quality and safety of egg products. Age-related variations in ultrastructure were thought to induce this deterioration. Eggshell formation is a highly complex process under precise regulation of genes and biological pathways in uterus of laying hens. Herein, we evaluated the physical, mechanical and ultrastructure properties of eggshell and conducted RNA sequencing to learn the transcriptomic differences in uterus between laying hens in the peak (young hens) and late phase (aged hens) of production.ResultsThe declined breaking strength and fracture toughness of eggshell were observed in aged hen group compared to those in young hen group, accompanied with ultrastructure variations including the increased thickness of mammillary layer and the decreased incidence of early fusion. During the initial stage of eggshell formation, a total of 183 differentially expressed genes (DEGs; 125 upregulated and 58 downregulated) were identified in uterus of laying hens in the late phase in relative to those at peak production. The DEGs annotated to Gene Ontology terms related to antigen processing and presentation were downregulated in aged hens compared to young hens. The contents of proinflammatory cytokine IL-1β in uterus were higher in aged hens relative to those in young hens. Besides, the genes of some matrix proteins potentially involved in eggshell mineralization, such as ovalbumin, versican and glypican 3, were also differentially expressed between two groups.ConclusionsAltered gene expression of matrix proteins along with the compromised immune function in uterus of laying hens in the late phase of production may conduce to age-related impairments of eggshell ultrastructure and mechanical properties. The current study enhances our understanding of the age-related deteriorations in eggshell ultrastructure and provides potential targets for improvement of eggshell quality in the late laying period.

Probiotic Clostridium butyricum could affect lipid metabolism in broilers. However, it is not clear whether C. butyricum could improve lipid metabolism through shaping gut microbiota and bile acid (BA) profile of laying hens. We aimed to evaluate the contributions of gut microbiota and BA profile to the potential effect of C. butyricum on lipid metabolism of aged laying hens. A total of 192 60-week-old Hy-Line Brown laying hens were divided into two groups (eight replicates per group). Birds were fed a basal diet supplemented with 0 or 2.7 g/kg C. butyricum (1.0 × 109 CFU/g). Samples were collected at the end of week 8 of the experiment. The results showed elevated (P < 0.05) concentrations of glucagon-like peptide 1, insulin and thyroid hormones in serum responded to C. butyricum addition, which also decreased (P < 0.05) hepatic free fatty acids contents, as well as increased (P < 0.05) the expression of hepatic acyl-CoA oxidase, farnesoid X receptor (FXR) and PPARα. C. butyricum addition increased (P < 0.05) Bacteroidetes abundance but tended to decrease (P < 0.10) Firmicutes abundance in the ileum. Besides, C. butyricum addition resulted in higher (P < 0.05) abundances of Clostridia (Clostridiales) and Prevotellaceae, concurrent with an increasing trend (P < 0.10) of Bifidobacteriaceae abundance and decreased the abundances of several harmful bacteria such as Klebsiella (P < 0.05). Regarding ileal BA profile, there was a reduced (P < 0.05) content of tauro-α-muricholic acid, increased (P < 0.05) contents of tauroursodeoxycholic acid and lithocholic acid, along with increasing trends (P < 0.10) of glycochenodeoxycholic acid and hyodeoxycholic acid contents due to C. butyricum addition, which also increased (P < 0.05) ileal FXR expression. Collectively, supplemental C. butyricum accelerated hepatic fatty acid oxidation, and shaped gut microbiota and BA profile, thus reducing fat deposition in the liver of aged laying hens.

BackgroundThe compromised performance of laying hens in the late phase of production relative to the peak production was thought to be associated with the impairment of intestinal functionality, which plays essential roles in contributing to their overall health and production performance. In the present study, RNA sequencing was used to investigate differences in the expression profile of intestinal functionality-related genes and associated pathways between laying hens in the late phase and peak phase of production.ResultsA total of 104 upregulated genes with 190 downregulated genes were identified in the ileum (the distal small intestine) of laying hens in the late phase of production compared to those at peak production. These upregulated genes were found to be enriched in little KEGG pathway, however, the downregulated genes were enriched in the pathways of PPAR signaling pathway, oxidative phosphorylation and glutathione metabolism. Besides, these downregulated genes were mapped to several GO clusters in relation to lipid metabolism, electron transport of respiratory chain, and oxidation resistance. Similarly, there were lower activities of total superoxide dismutase, glutathione S-transferase and Na+/K+-ATPase, and reductions of total antioxidant capacity and ATP level, along with an elevation in malondialdehyde content in the ileum of laying hens in the late phase of production as compared with those at peak production.ConclusionsThe intestine of laying hens in the late phase of production were predominantly characterized by a disorder of lipid metabolism, concurrent with impairments of energy production and antioxidant property. This study uncovers the mechanism underlying differences between the intestinal functionality of laying hens in the late phase and peak phase of production, thereby providing potential targets for the genetic control or dietary modulation of intestinal hypofunction of laying hens in the late phase of production.

World egg production and consumption have been increasing for the past decades. Traditional strategies in poultry nutrition have made vital contributions to this great growth in quantity. However, current global issues should be considered in modern egg production such as growing populations and food security, food safety and quality, limited resources and environmental problems. The development of knowledge of poultry nutrition and modern biotechnology provides novel nutritional approaches to closely fit the requirement of pullets and laying hens, which will consequently decrease the nutrition excretion and maintain the lower cost of feed. Nutrition has also been widely accepted as a strategy to influence health and diseases of laying hens. The maintenance of good health is an important prerequisite for improving productivity and egg quality. In addition, there are many measures and strategies for minimizing the incidence of egg defects and providing a choice of lifestyle to enhance human health. This paper reviews current research progress on developing innovative technologies and strategies to maximize animal health and performance, improve the quality of egg products and minimize pollution caused by poultry production.

Objective This study was conducted to evaluate the effects of dietary protein sources (soybean meal, SBM; low-gossypol cottonseed meal, LCSM; double-zero rapeseed meal, DRM) on laying performance, egg quality, and plasma parameters of laying hens. Methods A total of 432 32-wk-old laying hens were randomly divided into 6 treatments with 6 replicates of 12 birds each. The birds were fed diets containing SBM, LCSM100, or DRM100 individually or in combination with an equal amount of crude protein (CP) (LCSM50, DRM50, and LCSM50-DRM50). The experimental diets, which were isocaloric (metabolizable energy, 11.11 MJ/kg) and isonitrogenous (CP, 16.5%), had similar digestible amino acid profile. The feeding trial lasted 12 weeks. Results The daily egg mass was decreased in the LCSM100 and LCSM50-DRM50 groups (p0.05) and showed increased yolk color at the end of the trial (p0.05). Conclusion Together, our results suggest that the LCSM100 or DRM100 diets may produce the adverse effects on laying performance and egg quality after feeding for 8 more weeks. The 100.0 g/kg LCSM diet or the148.7 g/kg DRM diet has no adverse effects on laying performance and egg quality.

Dietary inclusions of baicalin and chlorogenic acid were beneficial for intestinal health in pigs. Nevertheless, it is unknown whether these plant-derived products had protection for intestine against bacterial challenge in chickens. This study was aimed at evaluating the potential mitigating effects of plant extracts (PE) from Flos lonicerae combined with Baikal skullcap (the active components are chlorogenic acid and baicalin) on intestinal disruption and dysbacteriosis induced by Salmonella pullorum in laying hens. A total of 216 41-week-old layers were randomly divided into 3 groups (6 replicates per group): negative control (NC), S. pullorum-infected positive control (PC), and the S. pullorum-infected group with supplementation of PE at 1000 mg/kg. All birds except those in NC were challenged with S. pullorum at the end of 4 weeks of the experiment. S. pullorum challenge impaired (P < 0.05) the production performance (egg production, feed intake, and feed efficiency) of laying hens, increased (P < 0.05) serum endotoxin content and frequency of Salmonella-positive organs, as well as up-regulated (P < 0.05) ileal expression of pro-inflammatory cytokines including IFNG, TNFA, IL8, and IL1B, whereas PE addition reversed (P < 0.05) these changes and increased (P < 0.05) ileal IL10 expression. Supplemental PE moderated ileal microbiota dysbiosis in challenged birds, characterized by a reduced abundance of Firmicutes along with increased abundances of Bacteroidetes (Bacteroides), Deferribacteres and several butyrate-producers such as Prevotellaceae, Faecalibacterium, Blautia, Butyricicoccus, Lachnoclostridium, and Olsenella, which may assist with energy harvesting and boost anti-inflammatory capacity of host. The decreased abundance of Firmicutes with the increased abundance of Bacteroidetes caused by PE addition had positive correlations with the decreased expression of ileal pro-inflammatory cytokines. The increased abundances of Bacteroidetes (Bacteroides) and Prevotellaceae following PE addition were also positively correlated with the improvement of performance (egg production and feed intake) of laying hens. Collectively, supplemental PE from Flos lonicerae in combination with Baikal skullcap alleviated S. pullorum-induced intestinal disruption and performance impairment in laying hens, which could be at least partially responsible by the modulation of gut microbial composition.

Dietary inclusions of baicalin and chlorogenic acid were beneficial for intestinal health in pigs. Nevertheless, it is unknown whether these plant-derived products had protection for intestine against bacterial challenge in chickens. This study was aimed at evaluating the potential mitigating effects of plant extracts (PE) from Flos lonicerae combined with Baikal skullcap (the active components are chlorogenic acid and baicalin) on intestinal disruption and dysbacteriosis induced by Salmonella pullorum in laying hens. A total of 216 41-week-old layers were randomly divided into 3 groups (6 replicates per group): negative control (NC), S. pullorum-infected positive control (PC), and the S. pullorum-infected group with supplementation of PE at 1000 mg/kg. All birds except those in NC were challenged with S. pullorum at the end of 4 weeks of the experiment. S. pullorum challenge impaired (P < 0.05) the production performance (egg production, feed intake, and feed efficiency) of laying hens, increased (P < 0.05) serum endotoxin content and frequency of Salmonella-positive organs, as well as up-regulated (P < 0.05) ileal expression of pro-inflammatory cytokines including IFNG, TNFA, IL8, and IL1B, whereas PE addition reversed (P < 0.05) these changes and increased (P < 0.05) ileal IL10 expression. Supplemental PE moderated ileal microbiota dysbiosis in challenged birds, characterized by a reduced abundance of Firmicutes along with increased abundances of Bacteroidetes (Bacteroides), Deferribacteres and several butyrate-producers such as Prevotellaceae, Faecalibacterium, Blautia, Butyricicoccus, Lachnoclostridium, and Olsenella, which may assist with energy harvesting and boost anti-inflammatory capacity of host. The decreased abundance of Firmicutes with the increased abundance of Bacteroidetes caused by PE addition had positive correlations with the decreased expression of ileal pro-inflammatory cytokines. The increased abundances of Bacteroidetes (Bacteroides) and Prevotellaceae following PE addition were also positively correlated with the improvement of performance (egg production and feed intake) of laying hens. Collectively, supplemental PE from Flos lonicerae in combination with Baikal skullcap alleviated S. pullorum-induced intestinal disruption and performance impairment in laying hens, which could be at least partially responsible by the modulation of gut microbial composition.

Genetic variant T329S in flavin-containing monooxygenase 3 (FMO3) impairs trimethylamine (TMA) metabolism in birds. The TMA metabolism that under complex genetic and dietary regulation, closely linked to cardiovascular disease risk. We determined whether the genetic defects in TMA metabolism may change other metabolic traits in birds, determined whether the genetic effects depend on diets, and to identify genes or gene pathways that underlie the metabolic alteration induced by genetic and diet factors. We used hens genotyped as FMO3 c.984 A>T as well as those with the homozygous normal genotype. For each genotype, hens were provided with either a corn-soybean meal basal diets (SM), which contains lower levels of TMA precursor, or the basal diets supplemented with 21% of rapeseed meal (RM), which contains higher levels of TMA precursor. An integrative analysis of metabolomic and transcriptomic was used to explore the metabolic patterns of FMO3 genetic variant in hens that were fed the two defined diets. In birds that consumed SM diets, the T329S mutation increased levels of plasma TMA and lipids, FMO3 mRNA levels, and the expression of genes involved in long chain polyunsaturated fatty acid biosynthesis. In birds that consumed RM diets, the T329S mutation induced fishy odor syndrome, a repression in LXR pathway and a reciprocal change in lipid metabolism. Variations in TMA and lipid metabolism were linked to the genetic variant in FMO3 in a diet-specific manner, which suggest FMO3 functions in TMA metabolism and lipid homeostasis. LXR pathway and polyunsaturated fatty acid metabolism are two possible mechanisms of FMO3 action in response to dietary TMA precursor.