Parental betaine supplementation promotes gosling growth with epigenetic modulation of IGF gene family in the liver.

Betaine is widely used as a feed additive in the chicken industry to promote laying performance and growth performance, yet it is unknown whether betaine can be used in geese to improve the laying performance of goose breeders and the growth traits of offspring goslings. In this study, laying goose breeders at 39 wk of age were fed basal (Control, CON) or betaine-supplemented diets at low (2.5 g/kg, LBT) or high (5 g/kg, HBT) levels for 7 wk, and the breeder eggs laid in the last week were collected for incubation. Offspring goslings were examined at 35 and 63 d of age. The laying rate tended to be increased (P = 0.065), and the feed efficiency of the breeders was improved by betaine supplementation, while the average daily gain of the offspring goslings was significantly increased (P < 0.05). Concentrations of insulin-like growth factor 2 (IGF-2) in serum and liver were significantly increased in the HBT group (P < 0.05), with age-dependent alterations of serum T3 levels. Concurrently, hepatic mRNA expression of the IGF gene family was significantly increased in goslings derived from betaine-treated breeders (P < 0.05). A higher ratio of proliferating cell nuclear antigen (PCNA)-immunopositive nuclei was found in the liver sections of the HBT group, which was confirmed by significantly upregulated hepatic expression of PCNA mRNA and protein (P < 0.05). Moreover, hepatic expression of thyroxine deiodinase type 1 (Dio1) and thyroid hormone receptor ß (TRß) was also significantly upregulated in goslings of the HBT group (P < 0.05). These changes were associated with significantly higher levels of global DNA 5-mC methylation, together with increased expression of methyl transfer genes (P < 0.05), including betaine-homocysteine methyltransferase (BHMT), glycine N-methyltransferase (GNMT), and DNA (cytosine-5-)-methyltransferase 1 (DNMT1). The promoter regions of IGF-2 genes, as well as the predicted TRß binding site on the IGF-2 gene, were significantly hypomethylated (P < 0.05). These results indicate that gosling growth can be improved by dietary betaine supplementation in goose breeders via epigenetic modulation of the IGF gene family, especially IGF-2, in the liver.

The goose industry plays important roles in economics, cultures, and ecosystems, yet the low laying and growth rates of many indigenous breeds hinders the development of the goose farming. Betaine, an important methyl donor, is commonly used as a feed additive in livestock and poultry to enhance animal growth. Dietary supplementation of betaine in laying hens or gestational sows has been reported to promote the growth of their offspring. Here, we sought to investigate whether and how dietary betaine supplementation affects the growth and development of offspring goslings. In this study, goose breeders, both male and female, were fed a basal diet supplemented respectively with 0, 2.5, or 5 g/kg betaine for 7 wk. Goslings hatched from the breeder eggs of different groups were raised under the same standard condition for assessing the growth performance. Parental betaine increases the growth rate of offspring goslings with decreased DNA methylation on the IGF-2 gene promoter and increased expression of the IGF-2 gene in the liver. These results provide scientific evidence for the inter-generational effect of betaine on gosling growth.

Exogenous betaine enhances salt tolerance of Glycyrrhiza uralensis through multiple pathways.

BACKGROUND: Glycyrrhiza uralensis Fisch., a valuable medicinal plant, shows contrasting salt tolerance between seedlings and perennial individuals, and salt tolerance at seedling stage is very weak. Understanding this difference is crucial for optimizing cultivation practices and maximizing the plant's economic potential. Salt stress resistance at the seedling stage is the key to the cultivation of the plant using salinized land. This study investigated the physiological mechanism of the application of glycine betaine (0, 10, 20, 40, 80 mM) to seedling stages of G. uralensis under salt stress (160 mM NaCl). RESULTS: G. uralensis seedlings' growth was severely inhibited under NaCl stress conditions, but the addition of GB effectively mitigated its effects, with 20 mM GB had showing most significant alleviating effect. The application of 20 mM GB under NaCl stress conditions significantly increased total root length (80.38%), total root surface area (93.28%), and total root volume (175.61%), and significantly increased the GB content in its roots, stems, and leaves by 36.88%, 107.05%, and 21.63%, respectively. The activity of betaine aldehyde dehydrogenase 2 (BADH2) was increased by 74.10%, 249.38%, and 150.60%, respectively. The 20 mM GB-addition treatment significantly increased content of osmoregulatory substances (the contents of soluble protein, soluble sugar and proline increased by 7.05%, 70.52% and 661.06% in roots, and also increased by 30.74%, 47.11% and 26.88% in leaves, respectively.). Furthermore, it markedly enhanced the activity of antioxidant enzymes and the content of antioxidants (SOD, CAT, POD, APX and activities and ASA contents were elevated by 59.55%, 413.07%, 225.91%, 300.00% and 73.33% in the root, and increased by 877.51%, 359.89%, 199.15%, 144.35%, and 108.11% in leaves, respectively.), and obviously promoted salt secretion capacity of the leaves, which especially promoted the secretion of Na+ (1.37 times). CONCLUSIONS: In summary, the exogenous addition of GB significantly enhances the salt tolerance of G. uralensis seedlings, promoting osmoregulatory substances, antioxidant enzyme activities, excess salt discharge especially the significant promotion of the secretion of Na+Future studies should aim to elucidate the molecular mechanisms that operate when GB regulates saline stress tolerance.

Concurrent enhancement of biomass production and phycocyanin content in salt-stressed Arthrospira platensis: A glycine betaine- supplementation approach.

In industrial-scale cultivation of microalgae, salinity stress often stimulates high-value metabolites production but decreases biomass yield. In this research, we present an extraordinary response of Arthrospira platensis to salinity stress. Specifically, we observed a significant increase in both biomass production (2.58 g L-1) and phycocyanin (PC) content (22.31%), which were enhanced by 1.26-fold and 2.62-fold, respectively, compared to the control, upon exposure to exogenous glycine betaine (GB). The biochemical analysis reveals a significant enhancement in carbonic anhydrase activity and chlorophyll a level, concurrent with reductions in carbohydrate content and reactive oxygen species (ROS) levels. Further, transcriptomic profiling indicates a downregulation of genes associated with the tricarboxylic acid (TCA) cycle and an upregulation of genes linked to nitrogen assimilation, hinting at a rebalanced carbon/nitrogen metabolism favoring PC accumulation. This work thus presents a promising strategy for simultaneous enhancement of biomass production and PC content in A. platensis and expands our understanding of PC biosynthesis and salinity stress responses in A. platensis.

Betaine supplementation alleviates corticosterone-induced hepatic cholesterol accumulation through epigenetic modulation of HMGCR and CYP7A1 genes in laying hens.

Excessive corticosterone (CORT) exposure could cause hepatic cholesterol accumulation in chickens and maternal betaine supplementation could decrease hepatic cholesterol deposition through epigenetic modifications in offspring chickens. Nevertheless, it remains uncertain whether providing betaine to laying hens could protect CORT-induced hepatic cholesterol accumulation via epigenetic mechanisms. This study aimed to examine the effects of dietary betaine on plasma and hepatic cholesterol contents, expression of cholesterol metabolic genes, as well as DNA methylation on their promoters in the liver of laying hens exposed to CORT. A total of 72 laying hens at 130 d of age were randomly divided into 3 groups: control (CON), CORT, and CORT+betaine (CORT+BET) groups. The experiment lasted for 35 d. Chickens in CON and CORT groups were fed a basal diet, whereas the CORT+BET group chickens were fed the basal diet supplemented with 0.1% betaine for 35 d. On d 28 of the experiment, chickens in CORT and CORT+BET groups received daily subcutaneous injections of CORT (4.0 mg/kg body weight), whereas the CON group chickens were injected with an equal volume of solvent for 7 d. The results showed that CORT administration led to a significant increase (P < 0.05) in the contents of cholesterol in plasma and liver, associated with activation (P < 0.05) of sterol regulatory element binding transcription factor 2 (SREBP2), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), lecithin-cholesterol acyltransferase (LCAT) and low-density lipoprotein receptor (LDLR) genes expression, and inhibition of cholesterol-7-alpha hydroxylase (CYP7A1) and sterol 27-hydroxylase (CYP27A1) genes expression in the liver compared to the CON. In contrast, CORT-induced up-regulation of HMGCR mRNA and protein abundances and downregulation of CYP7A1 mRNA and protein abundances were completely normalized (P < 0.05) by betaine supplementation. Besides, CORT injection led to significant hypomethylation (P < 0.05) on HMGCR promoter and hypermethylation (P < 0.05) on CYP7A1 promoter. Moreover, dietary betaine rescued (P < 0.05) CORT-induced changes in methylation status of HMGCR and CYP7A1 genes promoters. These results indicate that dietary betaine addition protects laying hens from CORT-induced hepatic cholesterol accumulation via epigenetic modulation of HMGCR and CYP7A1 genes.

Maternal methyl donor supplementation: A potential therapy for metabolic disorder in offspring.

The prevalences of diabetes mellitus and obesity are increasing yearly and has become a serious social burden. In addition to genetic factors, environmental factors in early life development are critical in influencing the prevalence of metabolic disorders in offspring. A growing body of evidence suggests the critical role of early methyl donor intervention in offspring health. Emerging studies have shown that methyl donors can influence offspring metabolism through epigenetic modifications and changing metabolism-related genes. In this review, we focus on the role of folic acid, betaine, vitamin B12, methionine, and choline in protecting against metabolic disorders in offspring. To address the current evidence on the potential role of maternal methyl donors, we summarize clinical studies as well as experimental animal models that support the impact of maternal methyl donors on offspring metabolism and discuss the mechanisms of action that may bring about these positive effects. Given the worldwide prevalence of metabolic disorders, these findings could be utilized in clinical practice, in which methyl donor supplementation in the early life years may reverse metabolic disorders in offspring and block the harmful intergenerational effect.

Protective influence of supplementary betaine against heat stress by regulating intestinal oxidative status and microbiota composition in broiler chickens.

To assess the impact of supplementing betaine (BT) under heat stress (HS) conditions on broiler performance and intestinal health from 21 to 42 days of age, a total of 150 male Ross 308 broilers were indiscriminately allotted to 3 treatments with 10 replications of 5 birds each. The control (CON) group was given a basal ration and accommodated at a thermoneutral condition (22 ± 1 °C), whereas the HS and HS + BT groups were raised under cyclic HS (33 ± 1 °C for 8 h and 22 ± 1 °C for 16 h per day) and received the basal ration without or with 1000 mg/kg BT, respectively. The HS reduced average daily gain (ADG); average daily feed intake; villus height (VH); VH to crypt depth (CD) ratio (VCR); activities of trypsin, lipase, glutathione peroxidase (GPX), and catalase; and enumeration of Lactobacillus and Bifidobacterium (P < 0.05) and augmented feed conversion ratio (FCR), CD, malondialdehyde (MDA) accumulation, and enumeration of Escherichia coli, Clostridium, and coliforms (P < 0.05). Conversely, BT supplementation heightened ADG, VH, VCR, trypsin activity, GPX activity, and populations of Lactobacillus and Bifidobacterium (P < 0.05) and lowered FCR, MDA accumulation, and Clostridium population (P < 0.05). Furthermore, the FCR value, trypsin and GPX activities, MDA content, and Bifidobacterium and Clostridium populations in the HS + BT group were nearly equivalent to those in the CON group. To conclude, feeding BT under HS conditions could improve broiler performance through improving intestinal health by specifically mitigating oxidative damage and enhancing the colonization of beneficial bacteria.

Effects of Acute Beetroot Juice and Sodium Nitrate on Selected Blood Metabolites and Response to Transient Ischemia: A Crossover Randomized Clinical Trial.

BACKGROUND: Modification of the nitrate (NO3)-nitrite (NO2)-nitric oxide (NO) pathway can be induced by oral intake of inorganic NO3 (NIT) or NO3-rich products, such as beetroot juice (BRJ). OBJECTIVES: The primary aim of this study was to evaluate the plasma changes in betaine, choline, trimethylamine (TMA), trimethylamine N-oxide (TMAO), and NO3/NO2 (NOx) concentrations over 4 h after single oral ingestion of NIT or BRJ. The flow-mediated skin fluorescence (FMSF) method was applied to measure the changes in nicotinamide adenine dinucleotide reduced form (NADH) in response to transient ischemia and reperfusion. We hypothesized that various sources of NO3 may differently affect endothelial and mitochondrial functions in healthy human subjects. METHODS: In a randomized crossover trial, 8 healthy young adults ingested 800 mg NO3 from either NIT or BRJ on 2 separate days with ≥3 d apart. Venous blood samples were collected every hour, and FMSF determination was applied bihourly. RESULTS: Plasma betaine and choline concentrations peaked at 1 h after BRJ ingestion, and remained significantly higher than baseline values at all time points (P < 0.001 and P < 0.001, compared to preingestion values). Over time, BRJ was more effective in increasing NOx compared with NIT (fixed-trial effect P < 0.001). Baseline fluorescence decreased after both NIT and BRJ consumption (fixed-time effect P = 0.005). Transient ischemia and reperfusion response increased because of NO3 consumption (fixed-time effect P = 0.003), with no differences between trials (P = 0.451; P = 0.912; P = 0.819 at 0, 2, and 4 h, respectively). CONCLUSIONS: Acute ingestion of BRJ elevated plasma betaine and choline, but not TMA and TMAO. Moreover, plasma NOx levels were higher in the BRJ trial than in the NIT trial. Various sources of NO3 positively affected endothelial and mitochondrial functions. This trial was registered at clinicaltrials.gov as NCT05004935.

Coping with Water Stress: Ameliorative Effects of Combined Treatments of Salicylic Acid and Glycine Betaine on the Biometric Traits and Water-Use Efficiency of Onion (Allium cepa) Cultivated under Deficit Drip Irrigation.

Freshwater scarcity is a major global challenge threatening food security. Agriculture requires huge quantities of water to feed the ever-increasing human population. Sustainable irrigation techniques such as deficit drip irrigation (DDI) are warranted to increase efficiency and maximize yield. However, DDI has been reported to cause water stress in plants. The study aimed to investigate the influence of the exogenous application of salicylic acid alone (SA) or in combination with glycine betaine (GB) on the growth, yield quality, and water-use efficiency of onions under different DDI treatments (100%, 70%, and 40% field capacity (FC)). Spray treatments (sub-treatments) were as follows: T1: (distilled water), T2: (1.09 mM SA), T3: (1.09 mM SA + 25 mM GB), T4: (1.09 mM SA + 50 mM GB), and T5: (1.09 mM SA + 100 mM GB). Our results indicated that T2 slightly ameliorated the effects of water stress by improved plant heights, leaf number, pseudostem diameter, bulb quality, and nutrient content of onion bulbs, especially under the 70% FC treatment. However, T3 recorded the poorest results on leaf number, pseudostem diameter, and bulb quality under the 70% and 40% FC treatments. Generally, our results indicated that onions could tolerate moderate water stress (70% FC) without severely affecting the growth and yield of onion. In conditions where freshwater is a limiting factor, a DDI treatment of 40% FC is recommended.

Effects of dietary betaine and protected calcium butyrate supplementation on growth performance, blood biochemical status, and meat quality in growing Japanese quail (Coturnix coturnix Japonica).

This study aimed to evaluate the impact of betaine (Bet) and protected calcium butyrate (PCB) supplementation individually and together on the performance, carcass traits, blood biochemistry, and meat quality of growing Japanese quails ( Coturnix coturnix Japonica) from 1 to 42 days old. 144 one-day-old unsexed Japanese quails were randomly assigned to four dietary treatments with six replicates each. All birds were fed a maize-soybean meal diet for 42 days. The control group received no feed additives, while the treatment groups received 1.2 g/kg Bet, 1.0 g/kg PCB, or a combination of both in their diets. The results indicated that Bet and PCB supplementation individually and together did not differ performance, relative weights of heart, gizzard, proventriculus, bursa of Fabricius and pancreas, water holding capacity (WHC), cooking loss (CL), blood biochemical values except for glucose and triglyceride. Bet supplementation significantly increased relative liver weights, while PCB supplementation decreased glucose levels in serum. Moreover, carcass yield was increased and triglyceride value in blood serum, malondialdehyde (MDA), and the pH levels of breast meats both on the 1st and 30st day of post-mortem were decreased in all treatment groups. Therefore, based on these results, the combination of betaine and butyrate improves both carcass yield and meat quality in growing Japanese quails. More research is needed to determine the impact of betaine and butyrate on the structure of amino acids in meat, antioxidant enzyme activity, and the immune system in poultry.

Identification of glycine betaine as a host-derived molecule required for the vegetative proliferation of the protozoan parasite Perkinsus olseni.

Perkinsus olseni is an industrially significant protozoan parasite of Manila clam, Ruditapes philippinarum. So far, various media, based on Dulbecco's Modified Eagle Medium and Ham's F-12 nutrient mixture with supplementation of fetal bovine serum (FBS), have been developed to proliferate the parasitizing trophozoite stage of P. olseni. The present study showed that P. olseni did not proliferate in FBS-deficient Perkinsus broth medium (PBMΔF), but proliferated well in PBMΔF supplemented with tissue extract of host Manila clams, indicating that FBS and Manila clam tissue contained molecule(s) required for P. olseni proliferation. Preliminary characterization suggested that the host-derived molecule(s) was a heat-stable molecule(s) with a molecular weight of less than 3 kDa, and finally a single molecule required for the proliferation was purified by high-performance liquid chromatography processes. High-resolution electrospray ionization mass spectrometry and nuclear magnetic resonance analyses identified this molecule as glycine betaine (=trimethylglycine), and the requirement of this molecule for P. olsseni proliferation was confirmed by an assay using chemically synthesized, standard glycine betaine. Although glycine betaine was required for the proliferation of all examined Perkinsus species, supplementation of glycine betaine precursors, such as choline and betaine aldehyde, enhanced the proliferation of 4 Perkinsus species (P. marinus, P. chesapeaki, P. mediterraneus and P. honshuensis), but not of 2 others (P. olseni and P. beihaiensis). Thus, it was concluded that the ability to biosynthesise glycine betaine from its precursors varied among Perkinsus species, and that P. olseni and P. beihaiensis lack the ability required to biosynthesize glycine betaine for proliferation.

Betaine supplementation improves CrossFit performance and increases testosterone levels, but has no influence on Wingate power: randomized crossover trial.

BACKGROUND: Because betaine (BET) supplementation may improve muscular strength and endurance, it seems plausible that BET will also influence CrossFit performance (CF). PURPOSE: The aim of this study was to evaluate the effects of three weeks of BET supplementation on body composition, CF performance, muscle power in the Wingate anaerobic test (WAnT), and the concentrations of selected hormones. The secondary aims were to analyze the effectiveness of two different BET doses (2.5 and 5.0 g/d) and their interaction with the methylenetetrahydrofolate reductase (MTHFR) genotype. METHODS: The study was designed in a double-blinded randomized cross-over fashion. Forty-three CF practitioners completed the entire study. CF performance was measured using the Fight Gone Bad (FGB) workout and muscle power was evaluated in a 30-second WAnT. Body composition was determined by air-displacement plethysmography. Blood was drawn to assess hormone concentrations. The C677T single nucleotide polymorphism (rs180113) in the MTHFR gene was analyzed. RESULTS: FGB total improved with BET by 8.7 ± 13.6% (p < 0.001), but no significant changes were observed with placebo (- 0.4 ± 10.0%, p = 0.128). No changes were also observed in WAnT and body composition. After BET supplementation testosterone concentration increased by 7.0 ± 15.4% with BET (p = 0.046) (no change with placebo: 1.5 ± 19.6%, p = 0.884) but had no effect on concentrations of insulin-like growth factor or cortisol. Finally, there were no significant interactions between MTHFR genotype and BET dose in any outcome. CONCLUSIONS: BET supplementation may improve CF performance and increase testosterone concentration. However, there was no evidence of a difference between dosages (2.5 and 5.0 g/d) and MTHFR genotypes. The trial was registered on clinicaltrials.gov (NCT03702205) on 10 October 2018.

Betaine as an alternative feed additive to choline and its effect on performance, blood parameters, and egg quality in laying hens rations.

This research aimed to evaluate how using betaine levels as a choline substitute affects productive performance, egg quality parameters, fatty acids profile, and antioxidant status in laying hens. One hundred and forty brown chickens, 45 wk old, were divided into 4 groups, each group of 7 replicates with 5 chickens per replicate. The first group of diets with choline has control (A) 100% choline, the second group (B) 75% choline + 25% betaine, the third group (C) 50% choline + 50% betaine, and the fourth group (D) received 100% betaine. No significant effects were observed in final body weight (BW), body weight gain (BWG), egg production (EW), and feed intake (FI) for laying hens. In the diet in which betaine was replaced choline, egg mass (EM) and egg weight (EW) increased compared to the control group (P < 0.05). Also, after 12 wk of feeding, the egg quality parameters were not influenced; however, yolk color was increased significantly compared with the control group. Serum total cholesterol, LDL-lipoprotein, HDL-lipoprotein, triglyceride, glucose, aspartate transaminase (AST), and alanine transaminase (ALT) were not affected by replacing choline with betaine. Furthermore, liver malondialdehyde (MDA) content, yolk vitamin E, and fatty acid levels were not significantly affected by replacing choline with betaine. Moreover, hens fed betaine displayed an increased antibody titer of the Newcastle disease (ND) virus. EW and EM were increased by 3.50% and 5.43% in 100% betaine group (D) when compared to the control group. Isthmus weight was decreased by 48.28 % in 50% choline + 50% betaine group (C) when compared to the control group. ND was increased by 26.24% in 100% betaine group when compared to the control group. In conclusion, betaine supplementation positively affected productive performance, egg quality measurements, and immunity response in Bovans brown laying hens.

Effect of individual or combination of dietary betaine and glycine on productive performance, stress response, liver health, and intestinal barrier function in broiler chickens raised under heat stress conditions.

The current experiment was conducted to investigate the effect of individual or combination of dietary betaine (Bet) and glycine (Gly) on productive performance, stress response, liver health, and intestinal barrier function in broiler chickens raised under heat stress (HS) conditions. A total of four hundred twenty 21-d-old Ross 308 broiler chickens were randomly allotted to 1 of 5 dietary treatments with 7 replicates. Birds in treatment 1 were raised under the thermoneutral condition (TN; 23 ± 0.6°C). Birds in other 4 treatment groups were subjected to a cyclic HS by exposing them to 32 ± 0.9°C for 8 h/d (from 09:00 to 17:00 h) and 28 ± 1.2°C for the remaining time for 14 d. Birds were fed a basal diet in TN condition (TN-C) and one group in HS conditions (HS-C), whereas other birds raised under HS conditions were fed the basal diet supplemented with 0.20% Bet (HS-Bet), 0.79% Gly (HS-Gly), or their combination (0.20% Bet + 0.79% Gly; HS-Bet+Gly). Results indicated that birds in HS-Bet, HS-Gly, or HS-Bet+Gly treatment had higher (P < 0.05) final BW and BW gain, but lower (P < 0.05) feed conversion ratio (FCR) than those in HS-C treatment. However, values for improved final BW, BW gain, and FCR by dietary treatments were lower (P < 0.05) than those measured in TN-C treatment. Under HS conditions, birds in HS-Bet, HS-Gly, or HS-Bet+Gly treatment had lower (P < 0.05) heterophil to lymphocyte ratio than those in HS-C treatment. Birds in HS-Gly or HS-Bet+Gly treatment had higher (P < 0.05) villus height and goblet cell number than birds in HS-C treatment. Intestinal permeability was higher (P < 0.05) in all HS-treatment groups than in TN-C treatment, but it was not affected by dietary treatment. In conclusion, individual supplementation of 0.20% Bet or 0.79% Gly in diets alleviates the negative effect of HS in broiler chickens. However, the synergistic effect of the combination of 0.20% Bet and 0.79% Gly in broiler diets seems lower than expected.

Effects of low-energy diet supplemented with betaine on growth performance, nutrient digestibility, and serum metabolomic profiles in growing pigs.

Two experiments were carried out to evaluate the effects of betaine (BET) supplementation in diets with reduced net energy (NE) levels on growth performance, nutrient digestibility, and serum metabolomic profiles in growing pigs. In experiment 1, 24 growing pigs (initial body weight, BW, 30.83 ±â€…2.50 kg) were allotted to one of the four treatments (six replications with 1 pig per pen) in a 2 × 2 factorial arrangement, including two dietary NE levels (2475 [N-NE] or 2395 [R80-NE] kcal/kg) and two BET doses (0 or 1500 mg/kg). In experiment 2, 72 growing pigs were used in a 2 × 3 factorial arrangement, including three dietary NE levels (2475 [N-NE], 2415 [R60-NE], or 2355 [R120-NE] kcal/kg) and two BET doses (0 or 1500 mg/kg). Pigs with initial BW of 31.44 ±â€…1.65 kg were divided to one of the six treatments (six replications with 2 pigs per pen). In experiment 1, lowing NE concentrations increased average daily feed intake (ADFI) by 10.69% in pigs fed the diet without BET (P > 0.05). BET significantly increased ADFI in N-NE diet (P < 0.05) but had no influence on ADFI in R80-NE diet (P > 0.05). BET enhanced the apparent digestibility of crude protein (CP), dry matter (DM), organic matter (OM), gross energy (GE), and ether extract (EE) in R80-NE diet (P < 0.05). In experiment 2, lowing NE concentrations enhanced ADFI (P > 0.05) and decreased average daily gain (ADG; P < 0.05). The reduction in feed intake by BET was further enhanced as NE concentrations decreased from 2415 to 2355 kcal/kg (P < 0.10). BET reversed the elevation of serum triglyceride, alkaline phosphatase, aspartate aminotransferase, and alanine aminotransferase levels caused by R120-NE diet (P < 0.05). The concentrations of cholecystokinin and glucagon-like peptide 1 were increased by BET in pigs fed the R120-NE diet (P < 0.05). Serum metabolomics reveals that lowing dietary NE concentrations affected mainly amino acid biosynthetic pathways (P < 0.05). BET supplementation in R120-NE diet up-regulated serum BET levels and down-regulated homocysteine, DL-carnitine, and four amino acid secondary metabolites (P < 0.05). In conclusion, lowing dietary NE contents reduced the growth performance and caused metabolic abnormalities in growing pigs. However, BET decreased feed intake to a certain extent and improved the metabolic health of pigs fed the low-NE diets, which may be related to the dual regulation of amino acid metabolism and the secretion of appetite related hormones by BET.

Energy is an important factor in affecting the production efficiency and feed cost in animal husbandry. For pigs, the reduction of dietary energy will lead to a decreased growth performance. Therefore, additional researches towards ameliorating the negative effects caused by low energy diets are necessary to conduct, so as to develop appropriate nutritional strategies. Betaine, a trimethyl derivative of glycine, is considered to affect energy partitioning. Betaine may influence the growth performance and healthy status of pigs under low-energy conditions. Herein, two experiments were carried out to evaluate the effects of betaine supplementation in diets with reduced net energy levels on growth performance, nutrient digestibility, and serum metabolomic profiles in growing pigs. Results indicated that lowering dietary energy reduced growth performance and caused metabolic abnormalities in growing pigs, however, betaine supplementation in low-energy diets improved metabolic homeostasis and the utilization of energy despite reduced feed intake to a certain extent.

Ascophyllum nodosum polysaccharide regulates gut microbiota metabolites to protect against colonic inflammation in mice.

Ascophyllum nodosum polysaccharide (ANP) can protect against colonic inflammation but the underlying mechanism is still unclear. This study has determined the metabolites of gut microbiota regulated by ANP to reveal the mechanism of the anti-inflammation effect of ANP. Using an in vitro colonic fermentation model, the results indicate that gut microbiota could utilize a proportion of ANP to increase the concentrations of short-chain fatty acids (SCFAs) and decrease ammonia content. Metabolomics revealed that 46 differential metabolites, such as betaine, L-carnitine, and aminoimidazole carboxamide ribonucleotide (AICAR), could be altered by ANP. Metabolic pathway analysis showed that ANP mainly up-regulated the phenylalanine, tyrosine, and tryptophan biosynthesis and aminoacyl-tRNA biosynthesis, which were negatively correlated with inflammation progression. Interestingly, these metabolites associated with inflammation were also up-regulated by ANP in colitis mice, including betaine, L-carnitine, AICAR, N-acetyl-glutamine, tryptophan, and valine, which were mainly associated with amino acid metabolism and aminoacyl-tRNA biosynthesis. Furthermore, the metabolites modulated by ANP were associated with the relative abundances of Akkermansia, Bacteroides, Blautia, Coprobacillus, Enterobacter, and Klebsiella. Additionally, based on VIP values, betaine is a key metabolite after the ANP supplement in vitro and in vivo. As indicated by these findings, ANP can up-regulate the production of SCFAs, betaine, L-carnitine, and AICAR and aminoacyl-tRNA biosynthesis to protect against colonic inflammation and maintain intestinal health.

Effects of emulsifier, betaine, and L-carnitine on growth performance, immune response, gut morphology, and nutrient digestibility in broiler chickens exposed to cyclic heat stress.

1. This experiment investigated the efficacy of varying doses of an emulsifier blend (EB; 0 and 1 g/kg of diet), betaine (BT; 0 and 1 g/kg of diet) and L-carnitine (CT; 0 and 0.5 g/kg of diet) in broilers subjected to circular heat stress (HS) conditions. A total of 1080 one-day-old male broiler chickens (Ross 308) were randomly assigned to one of nine treatment groups (six pens/treatment with 20 birds/pen) according to a completely randomised design. The thermoneutral control broiler chickens were housed at a comfortable temperature and fed a standard diet (no additives). The other eight groups were exposed to cyclic HS conditions (34°C) for 8 h (10:00-18:00).2. There were EB × BT × CT interactions for body weight (BW) at 24 d (P = 0.038) and average daily gain (ADG) during the 10-24 d period (P = 0.049), with the greatest values found with concurrent supplementation of three supplements.3. Inclusion of EB resulted in greater (P < 0.05) BW, ADG, European performance index, uniformity rate, primary antibody titres against sheep red blood cells (SRBC), duodenal villus height (VH) and villus surface area, digestible energy (DE) and the coefficient of apparent ileal digestibility (CAID) of dry matter, crude protein, and fat However, feed conversion ratio, mortality rate and heterophile to lymphocyte ratio were lower (P < 0.05).4. Dietary BT supplementation improved (P < 0.05) all performance indicators, primary antibody titres against SRBC and Newcastle disease virus, serum total antioxidant capacity, duodenal VH, Jejunal VH/crypt depth and the CAID of dry matter and crude protein. The effect of dietary supplementation with CT was limited to an increase (P < 0.05) in ADG (d 10-24) and a decrease (P < 0.05) in serum malondialdehyde concentration (42 d) and jejunal crypt depth (42 d).5. In conclusion, dietary supplementation of either EB or BT alone or in combination ameliorated some of the detrimental effects of HS on growth performance, immunity and intestinal health in broilers, while a minor positive effect on performance and antioxidant status was observed with CT supplementation.

Maternal Betaine Supplementation Mitigates Maternal High Fat Diet-Induced NAFLD in Offspring Mice through Gut Microbiota.

Maternal betaine supplementation has been proven to alleviate non-alcoholic fatty liver disease (NAFLD) in offspring caused by maternal high-fat diet (MHFD). The gut-liver axis plays an important role in NAFLD pathogenesis. However, whether maternal betaine supplementation can alleviate NAFLD in offspring by the gut-liver axis is unknown. C57BL/6J mice were fed with high-fat diet for 4 weeks before mating, and supplemented with 1% betaine during pregnancy and lactation. After weaning, offspring mice were fed with standard diet to 10 weeks. Maternal betaine supplementation reduced hepatic triglyceride content and alleviated hepatic steatosis in offspring mice exposed to MHFD. Furthermore, the mRNA expression of PPARα, CPT1α and FATP2 was increased and TNFα was reduced by maternal betaine supplementation. Maternal betaine intake decreased the relative abundances of Proteobateria, Desulfovibrio and Ruminococcus, but increased the relative abundances of Bacteroides and Parabacteroides. Moreover, maternal betaine intake increased the concentrations of short-chain fatty acids (SCFAs), including acetic acid, butyric acid and valeric acid, in the feces. Gut microbiota and SCFAs were significantly correlated with hepatic triglyceride content and expression of the above genes. Maternal betaine intake had no effect on other gut microbiota-related metabolites (bile acid and trimethylamine-n-oxide). Altogether, maternal betaine supplementation ameliorated MHFD-induced NAFLD possibly through regulating gut microbiota and SCFAs in offspring mice.

Effects of betaine supplementation on reproductive performance of breeding geese.

1. An experiment feeding three concentrations of betaine was conducted using breeding geese to analyse the reproductive performance, serum biochemical indexes, egg quality and intestinal immunity.2. A total of 450 female and 90 male Jiangnan White breeding geese were divided into three treatments, with five pen replicates each containing 30 female geese and 6 male geese.3. The results showed that there was no significant effect on the reproductive performance, serum biochemical indexes or jejunal villi goblet cells of geese with different levels of betaine in the diet (P > 0.05). Compared with the control group, the addition of 2.5 g/kg betaine to the diet showed a tendency to increase egg mass (P > 0.05) the betaine content in the yolk (P < 0.05). Feeding betaine significantly increased the height of jejunal villi and egg yolk total cholesterol content in female geese (P < 0.05).4. In conclusion, adding betaine to the goose diet was effective in its ability to improve intestinal structures. Adding 2.5 g/kg betaine to feed significantly increased the content of TCHOL and betaine in goose eggs.

Comparing effects of natural betaine and betaine hydrochloride on gut physiology in broiler chickens.

Betaine is a well-known component of poultry diets with various effects on nutritional physiology. For example, increased water retention due to the osmolytic effect of betaine increases the volume of the cell, thereby accelerating the anabolic activity, integrity of cell membrane, and overall performance of the bird. Betaine is a multifunctional component (trimethyl derivative) acting as the most efficient methyl group donor and as an organic osmolyte, which can directly influence the gastrointestinal tract integrity, functionality, and health. So far, nothing is known about the effect of betaine on the intestinal barrier in chickens. In addition, little is known about comparing natural betaine with its synthetic form. Therefore, an animal study was conducted to ascertain the effects of betaine supplementation (natural and synthetic) on performance and intestinal physiological responses of broilers. One hundred and five 1-day-old broiler chicks were randomly assigned into 3 groups with 35 birds each: control, natural betaine (1 kg active natural (n)-betaine/ton of feed) and synthetic (syn)-betaine-HCL (1 kg active betaine /ton of feed). Histological assessment showed lower jejunal crypt depth and villi height/crypt depth ratio in syn-betaine-HCL group compared with natural n-betaine fed birds. Furthermore, it was found that syn-betaine-HCL negatively affects the integrity of the intestine by increasing the intestinal paracellular permeability in both jejunum and cecum as evidenced by a higher mannitol flux. Additionally, syn-betaine-HCl significantly upregulated the IFN-γ mRNA expression at certain time points, which could promote intestinal permeability, as it plays an important role in intestinal barrier dysfunction. Body weight (BW) and body weight gain (BWG) did not differ (P > 0.05) between the control birds and birds supplemented with syn-betaine-HCL. However, the BW and BWG were significantly (P < 0.05) improved by the dietary inclusion of n-betaine compared with other treatments. Altogether, the dietary inclusion of n-betaine had a positive effect on performance and did not negatively affect gut paracellular permeability. Furthermore, our results show that syn-betaine-HCl induces changes in the intestine, indicating an alteration of the intestinal histology and permeability. Thus, natural or synthetic betaine has different effects, which needs to be considered when using them as a feed supplement.

Concurrent betaine administration enhances exercise-induced improvements to glucose handling in obese mice.

BACKGROUND AND AIMS: Betaine supplementation has been shown to enhance hepatic lipid metabolism in obese mice and improve exercise performance in healthy populations. We examined effects of betaine supplementation, alone or in combination with treadmill exercise, on the metabolic consequences of high fat diet (HFD)-induced obesity in mice. METHODS AND RESULTS: Male C57BL/6 J mice were fed chow or HFD. After 15 weeks, HFD mice were split into: HFD, HFD with betaine (1.5% w/v), HFD with treadmill exercise, and HFD with both betaine and exercise (15 m/min for 45min, 6 days/week; n = 12/group) for 10 weeks. Compared to HFD mice, body weight was significantly reduced in exercise and exercise-betaine mice, but not in mice given betaine alone. Similarly, adiposity was reduced by exercise but not by betaine alone. HFD-induced glucose intolerance was slightly improved by exercise, but not with betaine alone. Significantly greater benefits were observed in exercise-betaine mice, compared to exercise alone, such that GTT-outcomes were similar to controls. This was associated with reduced insulin levels during ipGTT, suggesting enhanced insulin sensitivity. Modest benefits were observed in fatty acid metabolism genes in skeletal muscle, whilst limited effects were observed in the liver. HFD-induced increases in hepatic Mpc1 (mitochondrial pyruvate carrier 1) were normalized by all treatments, suggesting potential links to altered glucose metabolism. CONCLUSIONS: Our data show that drinking 1.5% betaine was sufficient to augment metabolic benefits of exercise in obese mice. These processes appear to be facilitated by altered glucose metabolism, with limited effects on hepatic lipid metabolism.