Effect of dexamethasone on gene expression of cannabinoid receptor type 1 and adenosine monophosphate-activated protein kinase in the hypothalamus of broilers (Gallus domesticus).

Hypothalamic neural circuits play a critical role in integrating peripheral signals and conveying information about energy and nutrient status. We detected cannabinoid receptor type 1 (CB1) distribution in the hypothalamus, liver, duodenum, jejunum, and ileum among 7- and 35-day-old broilers. The effects of dexamethasone (DEX) on CB1 gene expression were evaluated by in vitro and in vivo experiments on glucocorticoid receptor (GR) and adenosine monophosphate-activated protein kinase (AMPK) in the hypothalamus of broilers. In vitro, hypothalamic cells from 17-day-old broiler embryos were incubated with either 0.1% dimethyl sulfoxide or DEX (100 nmol/mL) for 1 h. In the in vivo study, 28-day-old broilers were injected with DEX for 24 h or 72 h. Results showed that CB1 was mainly expressed in the hypothalamus, and 72 h DEX treatment increased the expression. One-day treatment of broilers with DEX did not change the hypothalamic CB1 gene expression. Moreover, DEX treatment for 24 h and 72 h increased the mRNA level of hypothalamic AMPKα2 and GR. However, no differences were observed on the gene expression of CB1, GR, and AMPKα2 in hypothalamic cells with DEX-treated for 1 h. In conclusion, CB1 is mainly expressed in the hypothalamus of broilers; 72-h DEX exposure can regulate the CB1 system and AMPK signaling pathway of the broiler hypothalamus.

Effects of sex and fasting/refeeding on hepatic AMPK signaling in chickens (Gallus gallus).

The alpha-1 isoform of chicken AMPK situates on the Z-chromosome, in contrast, the other isoforms in birds and the mammalian AMPKα1 are located on the autosomes. The present study aimed to investigate the role of hepatic AMPK signaling in adaptation to nutritional status and the potential sex-specific response in chickens. Hepatic genes and proteins were compared between the two sexes immediately after hatching. From 20d of age, chicks from each sex received feed treatments: Control was fed ad libitum; Fasted was starved for 24 h; Refed was fed for 4 h after a 24 h fasting. As a result, hepatic AMPKα1 mRNA level in males was significantly higher at both ages compared to females, due to the presence of Z-chromosomes. However, this did not make this kinase "male-bias" as it was eventually compensated at a translational level, which was not reported in previous studies. The protein levels and activation of AMPKα were even lower in newly-hatched male compared to female chicks, accompanied with a higher FAS and SREBP-1 gene expressions. Accordingly, hepatic G6PC2 mRNA levels in males were significantly lower associated with lower plasma glucose levels after hatching. Fasting activated hepatic AMPK, which in turn inhibited gene expression of GS, FAS and SREBP-1, and stimulated the downstream G6PC2 in both sexes. These changes recovered after refeeding. In conclusion, AMPK plays a role in adaptation to nutritional environment for both sexes. The Z-linked AMPK did not exert a sex-specific signaling, due to a "translational compensation" of AMPKα1.

Alpha-lipoic acid impairs body weight gain of young broiler chicks via modulating peripheral AMPK.

In mammals, the AMP-activated protein kinase (AMPK) pathways in the central and peripheral tissues coordinately integrate inputs from multiple sources to regulate energy balance. The present study was aimed to explore the potential role of hepatic AMPK in the energy homeostasis of broiler chickens. Diets with 0, 0.05% or 0.1% alpha-lipoic acid (α-LA), a known AMPK inhibitor were provided to broiler chicks for 7days. As a result, α-LA supplementation decreased the relative growth rate of broiler chicks. Hepatic AMPKα2 mRNA levels were significantly upregulated by dietary α-LA, in concert with the increased phosphorylated AMPKα protein levels. In addition, hepatic FAS mRNA levels together with the malonyl-CoA to total CoA ester ratio were reduced by α-LA supplementation. Moreover, the hepatic phosphorylated glycogen synthase levels were increased resulting in a markedly decreased hepatic glycogen content. In conclusion, dietary α-LA supplementation decreased the in vivo hepatic glycogenesis and lipogenesis via stimulating hepatic AMPKα mRNA levels and the phosphorylated gene product. The stimulatory effect of α-LA on hepatic AMPK mRNA and pAMPKα protein levels together with our previous observations regarding its inhibitory effect on hypothalamic AMPK may have altered the energy balance and hence impaired body weight gain of broiler chicks.

Chenodeoxycholic acid reduces feed intake and modulates the expression of hypothalamic neuropeptides and hepatic lipogenic genes in broiler chickens.

Bile acids have recently become an emerging research hot spot in mammals due to their roles as metabolic regulators and molecular signatures controlling whole-body metabolic homeostasis. Such effects are still unknown in avian (non-mammalian) species. We, therefore, undertook this study to determine the effect of chenodeoxycholic acid (CDCA) on growth performance and on the expression of hypothalamic neuropeptides and hepatic lipogenic genes in broiler chickens. Chickens fed with diet-containing 0.1% or 0.5% CDCA for two weeks exhibited a significant and a dose dependent reduction of feed intake and body weight compared to the control (standard diet). These changes were accompanied with a significant decrease in plasma glucose levels at d10 and d15 post-treatment. At molecular levels, CDCA treatment significantly up-regulated the expression of feeding-related hypothalamic neuropeptides (NPY, AgRP, ORX, CRH, Ghrl, and MC1R) and down-regulated the hypothalamic expression of SOCS3. CDCA treatment also decreased the mRNA levels of key hepatic lipogenic genes (FAS, ACCα, ME, ATPcl, and SCD-1) and their related transcription factors SREBP-1/2 and PPARα. In addition, CDCA reduced the hepatic expression of FXR and the adipokine, visfatin, and adiponectin genes compared to the control. Together, our data provide evidence that CDCA alters growth performances in broilers and modulates the expression of hypothalamic neuropeptides and hepatic lipogenic and adipocytokine genes.

AMPK is involved in the differential neonatal performance of chicks hatching at different time.

We have recently reported that the hatching time may be in relation to the distinct neonatal performance of female chicks. The present study was aimed to investigate the potential involvement of AMPK, an energy sensor which plays a pivotal role in energy homeostasis, in the distinct performance of the spread of hatching time model. As a result, hypothalamic AMPKα1 isoform gene expression was significantly higher in the late hatcher as compared to that of their early counterparts, whereas the total and phosphorylated levels of AMPKα subunit did not differ between the three hatchers. The hypothalamic orexigenic NPY and AgRP mRNA levels were higher in the late hatchers as compared to the early, and that of the middle hatchers was at an intermediate level. However, the anorexigenic POMC and CRH was also higher expressed in the late hatchers as compared to the early hatchers. In the liver, AMPKα2 mRNA level and the phosphorylation ratio of AMPKα was significantly lower in the late hatchers, as compared to their early counterparts. The hepatic phosphorylated GS levels of the late and middle hatchers were lower than that of their early counterparts. The expression of hepatic FTO gene of the late hatchers was significantly higher than that of their early and middle counterparts. Taken together, AMPK may play a significant role in the different neonatal performance of the spread of hatching time model. The central and peripheral AMPK in late hatchers exhibited a pattern of higher energy intake and lower energy expenditure, which resulted in a faster post-hatch growth.

The effect of insulin on plasma glucose concentrations, expression of hepatic glucose transporters and key gluconeogenic enzymes during the perinatal period in broiler chickens.

Chickens have blood glucose concentrations that are twofold higher than those observed in mammals. Moreover, the insulin sensitivity seems to decrease with postnatal age in both broiler and layer chickens. However, little is known about the response of insulin on plasma glucose concentrations and mRNA abundance of hepatic glucose transporters 1, 2, 3, 8, 9 and 12 (GLUT1, 2, 3, 8, 9 and 12) and three regulatory enzymes of the gluconeogenesis, phosphoenolpyruvate carboxykinase 1 and 2 (PCK1 and 2) or fructose-1,6-biphosphatase 1 (FBP1) in chicks during the perinatal period. In the present study, broiler embryos on embryonic day (ED)16, ED18 or newly-hatched broiler chicks were injected intravenously with bovine insulin (1µg/g body weight (BW)) to examine plasma glucose response and changes in hepatic mRNA abundance of the GLUTs, PCK1 and 2 and FBP1. Results were compared with a non-treated control group and a saline-injected sham group. Plasma glucose levels of insulin-treated ED18 embryos recovered faster from their minimum level than those of insulin-treated ED16 embryos or newly-hatched chicks. In addition, at the minimum plasma glucose level seven hours post-injection (PI), hepatic GLUT2, FBP1 and PCK2 mRNA abundance was decreased in insulin-injected embryos, compared to sham and control groups, being most pronounced when insulin injection occurred on ED16.

Reduced protein availability by albumen removal during chicken embryogenesis decreases body weight and induces hormonal changes.

NEW FINDINGS: What is the central question of this study? Prenatal protein undernutrition by albumen removal in an avian model of fetal programming leads to long-term programming effects, but when do these effects first appear and are these programming effects regulated by the same candidate genes as in mammals? What is the main finding and its importance? The present results indicate that prenatal protein undernutrition by albumen removal induces phenotypical and hormonal changes in the early posthatch period, when the mismatch between the prenatal and postnatal environment first arises, but these changes are not accompanied by an altered gene expression of the selected candidate genes. Studies of the chicken offer a unique model for investigation of the direct effects of reduced prenatal protein availability by the partial replacement of albumen with saline in eggs at embryonic day 1 (albumen-deprived group). The results were compared with mock-treated sham chicks and non-treated control chicks. Although no differences in hatch weight were found, body weight and growth were reduced in the albumen-deprived chicks until 3 weeks of age. The feed intake of the albumen-deprived chicks, however, was increased compared with the control (day 13-21) and the sham chicks (day 16-18). In the albumen-deprived chicks, the ratio of thyroxine to 3,5,3'-triiodothyronine in the plasma was increased compared with the control chicks, whereas the plasma corticosterone level was increased only at day 7 compared with both other groups. The plasma glucose concentration and glucose tolerance were not affected by treatment. Several candidate genes previously associated with effects of prenatal protein deprivation in mammals were examined in the liver of newly hatched chicks. Gene expression of glycogen synthase 2, glycogen phosphorylase 1, peroxisome proliferator-activated receptor α and γ and glucocorticoid receptor was not affected by the treatment. In conclusion, reduction of prenatal protein availability led to differences in body weight and influenced hormones involved in metabolism and growth. Gene expression of the selected candidate genes was not altered, in contrast to mammals.

Transition of maternal dietary n-3 fatty acids from the yolk to the liver of broiler breeder progeny via the residual yolk sac.

The aim of the present study was to evaluate the transfer of maternal dietary fatty acids (FA) from the yolk to the developing offspring, with special emphasis on n-3 FA eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). Six hundred forty Ross 308 breeders were housed from 6 to 58 wk of age in 16 pens resulting in 4 replicates per dietary treatment. They were fed 1 of 4 diets: a basal diet, rich in n-6 FA (CON), or an n-3 FA enriched diet formulated to obtain an EPA/DHA ratio of 1/1 (EPA=DHA), 1/2 (DHA), or 2/1 (EPA). At 28, 43, and 58 wk of age, 20 eggs per treatment were collected and analyzed for FA composition. At these same breeder ages, 600 fertilized eggs per treatment were incubated. At hatch the residual yolks of 25 chicks per treatment were collected and analyzed for FA composition. At every hatch, 180 chicks per treatment were raised under standard conditions and livers were sampled at d 1, 14, 28, and 38 d for FA analysis. Concentrations of EPA in the yolk and residual yolk of eggs laid by EPA-fed breeders were highest, next-to-highest for EPA=DHA-fed breeders, next-to-lowest for DHA-fed breeders, and lowest in those laid by control hens, reflecting the inclusion levels in the maternal diets. Yolk and residual yolk DHA concentrations, however, were not only elevated due to DHA supplementation, compared with the control diet, but also due to EPA supplementation. Offspring hepatic EPA concentrations were elevated until d 28 in all n-3 enriched groups, whereas hepatic DHA concentrations were only affected by EPA=DHA and DHA supplementation at d 1. No differences were found in hepatic DHA concentrations at later offspring ages. Considering the role of EPA and DHA in early development and growth, the maternal supply of these n-3 FA might improve offspring health and performance.

Changes in oxidative stress in response to different levels of energy restriction in obese ponies.

The present study evaluated the effect of different levels of energy restriction on metabolic parameters in obese ponies. Relative weight changes, markers of lipid metabolism and oxidant/antioxidant balance were monitored. A total of eighteen obese (body condition score ≥ 7/9) Shetland ponies were studied over a 23·5-week trial, which was divided into three periods. The first period involved a 4-week adaptation period in which each animal was fed 100% of their maintenance energy requirements needed to maintain a stable obese body weight (MERob). This was followed by a 16·5-week weight-loss period in which ponies were assigned to receive either 100% (control group, CONTROL), 80% (slow weight-loss (SLOW) group) or 60% (rapid weight-loss (RAPID) group) of their MERob. During the 3-week end-phase period, all ponies were again fed 100% of their MERob. Relative weight loss was higher in the RAPID group (P< 0·001) compared with the SLOW group. No linear relationship was found as a doubling of the percentage of energy restriction was accompanied by a tripling of the percentage of weight loss. Relative weight gain afterwards in the end-phase period was higher in the RAPID group (P< 0·001) compared with the SLOW and CONTROL groups. During the weight-loss period, TAG and NEFA concentrations were highest in the RAPID group, as were α-tocopherol and ferric-reducing ability of plasma concentrations. After 8 weeks of weight loss, the concentrations of advanced oxidation protein products were higher in the RAPID group compared with the SLOW and CONTROL groups (P< 0·001). In conclusion, the level of energy restriction influences the extent of changes in oxidant/antioxidant balance. Practically, more severe energy restriction regimens may be associated with a greater regain of weight after the restriction period.

The effect of albumen removal before incubation (embryonic protein under-nutrition) on the post-hatch performance, regulators of protein translation activation and proteolysis in neonatal broilers.

Albumen was removed from broiler eggs before the start of incubation to induce prenatal protein under-nutrition in chicken embryos. With this method, the direct effect of protein deficiency was investigated, differing from mammalian models manipulating the maternal diet where indirect, hormonal effects can interfere. Based on the estimated albumen/egg weight ratio, 10 % of albumen was removed with an 18G needle, after making a hole at the sharp end of the egg with another 18G needle. Eggs were taped thereafter. The sham group underwent the same procedure, except that no albumen was removed. Control eggs did not receive any treatment. The removal of albumen decreased both embryonic and post-hatch body weight up to day 7 compared with the control group. On embryonic day 18, embryos from the albumen-deprived group had higher plasma uric acid levels compared with the sham (P= 0·016) and control (P= 0·009) groups. Moreover, a lower plasma amino acid concentration was observed at hatch compared with the sham (P= 0·038) and control (P= 0·152) groups. These findings indicate an altered protein metabolism. At hatch, a higher mRNA expression of muscle ring finger-1 (MuRF1), a gene related to proteolysis, was observed in albumen-deprived chicks compared with the control and sham chicks, together with an up-regulated expression of atrogin-1 (another atrogene) at this time point in the male protein-deficient chicks. These findings suggest that muscle proteolysis is transiently increased by the removal of albumen before the start of incubation. No evidence was found for altered protein synthesis capacity and translational efficiency in albumen-deprived chicks.

Effects of dietary arabinoxylan-oligosaccharides (AXOS) and endogenous probiotics on the growth performance, non-specific immunity and gut microbiota of juvenile Siberian sturgeon (Acipenser baerii).

We investigated the effects of administration of putative endogenous probiotics Lactococcus lactis spp. lactis or Bacillus circulans, alone and in combination with arabinoxylan-oligosaccharides (AXOS), a new class of candidate prebiotics, in juvenile Siberian sturgeon (Acipenser baerii). Eight experimental diets were tested: basal diet (Diet 1), basal diet supplemented with 2% AXOS (Diet 2), or L. lactis ST G81 (Diet 3), L. lactis ST G45 (Diet 4), B. circulans ST M53 (Diet 5), L. lactis ST G81 + 2% AXOS (Diet 6), L. lactis ST G45 + 2% AXOS (Diet 7), B. circulans ST M53 + 2% AXOS (Diet 8). After four weeks, growth performance and feed conversion ratio significantly improved in fish fed diet 7. Innate immune responses of fish were boosted with both AXOS and probiotic diets, however synergistic effects of AXOS and probiotic diets were only observed for phagocytic and alternative complement activity. Phagocytic and respiratory burst activity of fish macrophage increased in fish fed diet 2 and 7, while humoral immune responses only increased in fish fed diet 7. Pyrosequencing analysis (16S rDNA) of the hindgut microbiota demonstrated that AXOS improved the colonization or/and growth capacity of L. lactis, as a higher relative abundance of L. lactis was observed in fish receiving diet 7. However, no observable colonization of B. circulans was found in the hindgut of fish fed diet 5 or 8, containing this bacterium. The dietary L. lactis ST G45 + 2% AXOS caused significant alterations in the intestinal microbiota by significantly decreasing in bacterial diversity, demonstrated by the fall in richness and Shannon diversity, and improved growth performance and boosted immune responses of Siberian sturgeon.

The endocrine control of energy homeostasis in chickens.

Energy homeostasis (balance) depends on the relationship between the amount of consumed feed energy and energy expenditure. Coordination of energy expenditure and feed intake (appetite) is necessary for the regulation of body composition. The hypothalamus integrates peripheral and central signals to generate satiety or hunger. Birds and mammals utilize common signaling molecules but some molecules possess different/opposite functions. If relevant, particular differences with the mammalian regulatory system are highlighted in this review. For example, obestatin had no significant effect on feed intake of chicks, but it was claimed to decrease food intake in mammalian species. Ghrelin displayed appetite-stimulating effects in mammals but appetite-decreasing effects in birds. Recently, the function of the hypothalamic AMPK signaling pathway on feed intake regulation has received considerable attention in poultry. Alpha-lipoic acid might exert its appetite-decreasing effect by the AMPK signaling pathway. This review discusses the central regulation of energy homeostasis, role of (an)orexigenic peptides, effect of feed deprivation on hypothalamic neuropeptide gene expression and provides a model for involvement of AMPK in the regulation of avian energy balance.

Cerulenin upregulates heat shock protein-70 gene expression in chicken muscle.

Lines of evidence suggested that systems involved in the regulation of the stress responses and energy homeostasis are highly integrated. Because cerulenin, the natural antibiotic product of the fungus Cephalosporium ceruleans and a broad-spectrum fatty acid synthesis (FAS) inhibitor, has been shown to affect food intake and energy balance, and because the biomarker of stress Hsp-70 gene was found to interact directly with fatty acids, we hypothesized that cerulenin may regulate Hsp-70 gene expression. Therefore, the present study was undertaken to examine this issue. Cerulenin administration significantly (P < 0.05) decreased food intake and induced Hsp-70 mRNA levels in muscle, but not in liver or hypothalamus of 2-wk-old broiler chickens. These changes were accompanied by an unpregulation of muscle uncoupling protein and carnitine palmitoyltransferase 1 mRNA levels. This result indicated that the regulation of Hsp-70 gene expression in normal chickens, as estimated by oxidative stress indices [TBA reacting substances, ferric reducing/antioxidant power, and ceruloplasmin oxidase activity] levels, is tissue-specific. In attempt to discriminate between the effect of cerulenin and cerulenin-reduced food intake on Hsp-70 gene expression, we also evaluated the effect of food deprivation on the same cellular responses. Food deprivation for 16 h did not affect Hsp-70 gene expression in all tissues examined, indicating that the effect of cerulenin is independent of the inhibition of food intake. To ascertain whether the effect of cerulenin is direct or indirect, we carried out in vitro studies. Cerulenin treatment did not affect Hsp-70 gene expression in Leghorn male hepatoma and quail myoblast cell lines, suggesting that the observed effect in vivo may be mediated through the central nervous system.

Reduced-particle size wheat bran and endoxylanase supplementation in broiler feed affect arabinoxylan hydrolysis and fermentation with broiler age differently.

Since the caecal microbiota of young broilers are not yet able to ferment the dietary fibre (DF) fraction of the feed to a large extent, increasing the accessibility of DF substrates along the gastrointestinal tract is crucial to benefit from the health stimulating metabolic end-products (e.g. butyric acid) generated upon microbial DF fermentation. Therefore, the present study aimed to evaluate the potential of reduced-particle size wheat bran (RPS-WB) and endoxylanases as feed additives to stimulate arabinoxylan (AX) hydrolysis and fermentation along the hindgut of young broilers. To this end, RPS-WB and endoxylanase supplementation were evaluated in a 2 × 2 factorial design using a total of 256 male 1-d-old chicks (Ross 308). Broilers were assigned to 4 dietary treatments: a basal wheat-based diet with (1) no feed additives (control, CTRL), (2) an endoxylanase (XYL; Econase XT 25 at 0.10 g/kg diet), (3) 1% wheat bran with an average reduced particle size of 297 µm (RPS-WB) and (4) an endoxylanase and 1% RPS-WB (RPS-WB + XYL). Each dietary treatment was replicated 8 times and on d 10 and 28, respectively, 24 and 16 broilers per treatment group were euthanised to analyse AX degradation, short-chain fatty acid production and digesta viscosity in the ileum and caecum. Broilers receiving XYL in their diet showed increased AX solubilisation and fermentation at both d 10 and 28 compared to the CTRL group (P < 0.05). Adding RPS-WB to the diet stimulated wheat AX utilisation by the primary AX degraders in the caecum at 10 d of age compared to the CTRL group, as observed by the high AX digestibility coefficient for the RPS-WB supplemented group at this young age (P < 0.05). At 28 d, RPS-WB supplementation lowered body-weight gains but increased butyric acid concentrations compared to the XYL and CTRL group (P < 0.05). Although no synergistic effect for RPS-WB + XYL broilers was observed for AX hydrolysis and fermentation, these findings suggest that both additives can raise a dual benefit to the broiler as a butyrogenic effect and improved AX fermentation along the ileum and caecum were observed throughout the broiler's life.

Toward Renewable-Based Prebiotics from Woody Biomass: Potential of Tailored Xylo-Oligosaccharides Obtained by Enzymatic Hydrolysis of Beechwood Xylan as a Prebiotic Feed Supplement for Young Broilers.

Although antibiotic resistance emerges naturally, this process has been accelerated by the worldwide overuse and misuse of antibiotics. It is essential to find effective alternatives in the broiler industry to improve poultry health while maintaining production efficiency and product safety. In this study, we aimed to evaluate a potential alternative: wood-derived xylo-oligosaccharides (XOS). The objective of this research was to investigate the potential of XOS prepared using enzymatic hydrolysis of beechwood xylan as a prebiotic feed supplement for broilers. A pilot study was conducted to explore the optimal XOS fraction profile by in vitro fermentation. Subsequently, a semi-continuous enzyme membrane reactor was used, allowing for the production of tailored XOS in large quantities. Given the strong bidirectional relationship between intestinal health, nutrition, and intestinal microbiota composition in broilers, an in vivo experiment was performed to explore the potential of XOS as a prebiotic feed supplement by investigating growth performance, feed conversion ratio, caecal short and medium chain fatty acid (SCFA and MCFA) concentration, and microbiological composition of the caecal content. Results from the pilot study indicated that higher enzyme concentrations in the hydrolysis process yield a product that leads to a higher total SCFA and MCFA- and butyric acid production during in vitro fermentation by caecal bacteria. Supplementation of the tailored XOS to the broiler diet (day 1 (d1)-d8 0.13% wt/wt XOS, d9-d15 0.32% XOS) resulted in higher Bifidobacterium counts, beneficial to the health of birds, on d11 and d15.

Effects of arabinoxylan-oligosaccharides (AXOS) on juvenile Siberian sturgeon (Acipenser baerii) performance, immune responses and gastrointestinal microbial community.

Arabinoxylan-oligosaccharides (AXOS) are a newly discovered class of candidate prebiotics that exert different properties depending on their structure. In this study the effects of two different structures of AXOS, namely AXOS-32-0.30 (average degree of polymerization: 32, average degree of substitution: 0.30) and AXOS-3-0.25, were investigated on growth performance, immune responses, gut microbial fermentation and gut bacterial composition of juvenile Siberian sturgeon (Acipenser baerii). After a two weeks acclimation, fish (25.9 ± 0.9 g) were distributed over 24 aquariums (8 replicates per treatment) and fed a control diet or a diet containing 2% AXOS-32-0.30 or AXOS-3-0.25 for 12 weeks. Growth performance and feed utilization tend to improve in sturgeon fed on diets supplemented with AXOS-32-0.30, however not significant. Survival was high in all groups. Both AXOS preparations significantly enhanced the phagocytic activity of fish macrophages compared to the control group, while the alternative haemolytic complement activity and total serum peroxidase content improved only in the group fed AXOS-32-0.30 (P < 0.05). The lysozyme activity was not affected by AXOS addition. Simultaneously, the amount of short-chain fatty acids (SCFAs) was highest in the hind gut of sturgeon fed AXOS-32-0.30. The concentrations of acetate, butyrate and total SCFAs in fish fed AXOS-32-0.30 was significantly higher than in the groups fed the control diet or AXOS-3-0.25. Study of the bacterial community in the sturgeon hindgut using PCR-denaturing gradient gel electrophoresis (PCR-DGGE) revealed that both preparations of AXOS induced changes in the bacterial composition. According to redundancy analysis (RDA), hindgut microbiota of each treatment group clustered apart from one another (P = 0.001). DNA sequencing of the dominant DGGE bands recovered from the different treatments showed that AXOS mainly stimulated the growth of lactic acid bacteria and Clostridium sp., with more pronounced effects of AXOS-32-0.30. It is concluded that AXOS improves sturgeon health through prebiotic action, but the induced effects depend on the specific structure of AXOS. A higher degree of polymerization of AXOS had a stronger beneficial impact in this sturgeon species.

Fasting alters protein expression of AMP-activated protein kinase in the hypothalamus of broiler chicks (Gallus gallus domesticus).

An experiment was conducted to investigate the effects of fasting and re-feeding on hypothalamic 5'-AMP-activated protein kinase (AMPK) levels and (an)orexigenic neuropeptides. Male Arbor Acres chicks (7-day-old, n=160) were allocated to four equal treatment groups: control chicks (fed ad libitum for 48 h, C48), chicks that were fasted for 48 h (F48), chicks that were first fasted for 48 h and then re-fed for 24h (F48C24), and chicks that were fed ad libitum for 72h (C72). Fasting for 48 h significantly (P<0.05) increased the ratio of phosphorylated AMPKα to total AMPKα and phosphorylated LKB1 to total LKB1, whereas re-feeding for 24h reduced these ratios to that of the ad libitum fed C72 chicks. The gene expressions of agouti-related peptide (AgRP), neuropeptide Y (NPY), melanocortin receptor 4, melanin-concentrating hormone, prepro-orexins and carnitine palmitoyltransferase-1 were significantly (P<0.05) increased in the fasted chicks relative to the ad libitum fed C48 group. The gene expression of pro-opiomelanocortin (POMC), as well as cocaine- and amphetamine-regulated transcript (CART) was not affected by the nutritional status. Fasting significantly (P<0.05) decreased the mRNA levels of fatty acid synthase (FAS) and sterol regulatory element binding protein-1 (SREBP-1). The results suggest that the LKB1/AMPK signal pathway is involved in the energy homeostasis of fasted chicks, and its possible role in feed intake regulation might be mediated by the AgRP/NPY rather than the POMC/CART pathway.

Matrix metalloproteinase inhibition affects adipose tissue mass in obese mice.

1. Because the development of adipose tissue involves remodelling of the extracellular matrix (ECM), which requires matrix metalloproteinase (MMP) activity, we examined whether MMP inhibitors may have the potential to affect adipose tissue mass in obese mice. 2. Administration of the relatively gelatinase-specific MMP inhibitor tolylsam ((R)-3-methyl-2-[4-(3-p-tolyl-[1,2,4]oxadiazol-5-yl)-benzenesulphonylamino]-butyric acid; 100 mg/kg per day) for 7 weeks to obese wild-type mice on a high-fat diet resulted in significantly lower bodyweight (P < 0.05), lower subcutaneous (SC) and gonadal (GON) adipose tissue mass (both P < 0.05) and smaller adipocytes in both SC (P < 0.005) and GON (P < 0.0005) adipose tissues. 3. Magnetic resonance imaging confirmed a lower total body fat content in tolylsam-treated mice (P < 0.0005). In addition, tolylsam treatment of wild-type mice was associated with a marked enhancement in metabolic rate. 4. Electron microscopy analysis of tissue sections at the end of the 7 week feeding period revealed significantly higher collagen accumulation in the ECM of SC adipose tissues of tolylsam-treated mice (P < 0.001). 5. Thus, the relatively gelatinase-specific MMP inhibitor tolylsam has the potential to affect fat tissue growth in obese mice.

Chicken FTO gene: tissue-specific expression, brain distribution, breed difference and effect of fasting.

Fat mass and obesity-associated (FTO) gene is widely expressed in central and peripheral tissues of mammals, and exhibits a range of functions, especially in energy balance. However, basic knowledge of FTO in the chicken is lacking. Therefore, we studied the tissue distribution, age and breed dependent changes, brain localization, as well as the impact of fasting on FTO mRNA expression in the chicken. FTO mRNA was expressed in all the tissues studied, and generally, with high expression in hypothalamus, liver, visceral fat and cerebellum. However it exhibited breed-specific patterns: in broilers, the highest expression was seen in the liver, while in layers, hypothalamus and cerebellum showed relatively higher FTO mRNA expression. One-week-old broilers expressed markedly higher FTO mRNA in liver compared with the layers of the same age (P<0.01), while the breed difference was reversed in visceral fat and cerebellum (P<0.05). Compared with newly hatched chicks (one week of age), adult layers expressed higher FTO mRNA in liver and visceral fat, while adult broilers showed higher expression in hypothalamus and cerebellum. In situ hybridization demonstrated distribution of FTO mRNA in paraventricularis magnocellularis (PVN), nucleus ventromedialis hypothalami (VMN), nucleus lateralis hypothalami (LHy), nucleus dorsomedialis hypothalami (DMN) of the hypothalamus and nucleus habenularis medialis (HM) and stratum cellulare externum (SCE) of the thalamus. Breed-specific expression of FTO mRNA was shown in PVN, but not in VMN, with higher abundance in broilers compared to layers. The decrease in FTO mRNA levels after 24h of fasting was seen only in VMN of layer chickens. These results may provide some intriguing hints for further investigation of FTO function in the chicken.

A method for intracerebroventricular cannulation of young broiler chicks.

The site-specific administration of neuroactive substances or pharmacological agents is a routine practice in neurological studies. To facilitate chronic treatments intercranial cannulae are often implanted in the skull. These surgical procedures are widely described and well-documented for rodents, as the most common animal model, and even refined over the years. However, precise descriptions of proper procedure for third ventricle cannulation in young broiler chicks is not readily available. This absence is surprising, as chicken models are often used in the investigation of physiological control of the metabolism and ingestion. Furthermore, as a commercial animal, there is also a particular interest in elucidating the central regulation of feed intake and metabolism to optimize feeding and living conditions for broilers. The brain nuclei that serve as the regulatory centers of feeding and metabolism are the arcuate nucleus, the ventromedial hypothalamic nucleus, and the lateral hypothalamus, which are all located near the third ventricle. Here, we provide a full protocol for intracerebroventricular (i.c.v.) cannulation of 7-day old broiler chicks, as well as a review of some of the advances made in rodent i.c.v. cannulations and whether these advances are applicable to cannulations in chickens. Using the surgical procedure described here, we were able to achieve a success rate of proper implantation of 88%, with a mortality rate of less than 1% (n = 224).•Detailed procedure for i.c.v. canula implantation in the third ventricle of 7d-old broiler chicks.•Cement cap with anchoring screws is necessary in broilers, glue does not provide sufficient stability.•Carboxylate cement and self-adhesive resin cement were tested as an alternative for methyl methacrylate cement.