Effects of essential amino acids supplementation in a low-protein diet on growth performance, intestinal health and microbiota of juvenile blotched snakehead (Channa maculata).

Developing a low-protein feed is important for the sustainable advancement of aquaculture. The aim of this study was to investigate the effects of essential amino acid (EAA) supplementation in a low-protein diet on the growth, intestinal health, and microbiota of the juvenile blotched snakehead, Channa maculata in an 8-week trial conducted in a recirculating aquaculture system. Three isoenergetic diets were formulated to include a control group (48.66 % crude protein (CP), HP), a low protein group (42.54 % CP, LP), and a low protein supplementation EAA group (44.44 % CP, LP-AA). The results showed that significantly lower weight gain (WG), specific growth rate (SGR), protein efficiency ratio (PER), and feed efficiency ratio (FER) were observed in fish that were fed LP than in the HP and LP-AA groups (P < 0.05). The HP and LP-AA groups exhibited a significant increase in intestinal villus length, villus width, and muscular thickness compared to the LP group (P < 0.05). Additionally, the HP and LP-AA groups demonstrated significantly higher levels of intestinal total antioxidant capacity (T-AOC), catalase (CAT), and superoxide dismutase (SOD) and lower levels of malondialdehyde (MDA) compared to the LP group (P < 0.05). The apoptosis rate of intestinal cells in the LP group was significantly higher than those in the LP and HP groups (P < 0.05). The mRNA expression levels of superoxide dismutase (sod), nuclear factor kappa B p65 subunit (nfκb-p65), heat shock protein 70 (hsp70), and inhibitor of NF-κBα (iκba) in the intestine were significantly higher in the LP group than those in the HP and LP-AA groups (P < 0.05). The 16s RNA analysis indicated that EAA supplementation significantly increased the growth of Desulfovibrio and altered the intestinal microflora. The relative abundances of Firmicutes and Cyanobacteria were positively correlated with antioxidant parameters (CAT and T-AOC), whereas Desulfobacterota was negatively correlated with sod and T-AOC. The genera Bacillus, Bacteroides, and Rothia were associated with the favorable maintenance of gut health. In conclusion, dietary supplementation with EAAs to achieve a balanced amino acid profile could potentially reduce the dietary protein levels from 48.66 % to 44.44 % without adversely affecting the growth and intestinal health of juvenile blotched snakeheads.

Effects of rumen-encapsulated methionine and lysine supplementation and low dietary protein on nitrogen efficiency and lactation performance of dairy cows.

Low crude protein (CP) diets might be fed to dairy cows without affecting productivity if the balance of absorbed AA were improved, which would decrease the environmental effect of dairy farms. The aim of this study was to investigate the effects of supplementing ruminally protected Lys (RPL) and Met (RPM) at 2 levels of dietary CP on nutrient intake, milk production, milk composition, milk N efficiency (MNE), and plasma concentrations of AA in lactating Holstein cows and to evaluate these effects against the predictions of the new NASEM (2021) model. Fifteen multiparous cows were used in a replicated 3 × 3 Latin square design with 21-d periods. The 3 treatments were (1) a high-protein (HP) basal diet containing 16.4% CP (metabolizable protein [MP] balance of -130 g/d; 95% of target values), (2) a medium-protein diet containing 15% CP plus RPL (60 g/cow per day) and RPM (25 g/cow per day; MPLM; MP balance of -314 g/d; 87% of target values), and (3) a low-protein diet containing 13.6% CP plus RPL (60 g/cow per day) and RPM (25 g/cow per day; LPLM; MP balance of -479 g/d; 80% of target values). Dry matter intake was less for cows fed MPLM and LPLM diets compared with those fed the HP diet. Compared with the HP diet, the intake of CP, neutral detergent fiber, acid detergent fiber, and organic matter, but not starch, was lower for cows fed MPLM and LPLM diets. Milk production and composition were not affected by MPLM or LPLM diets relative to the HP diet. Milk urea N concentrations were reduced for the MPLM and LPLM diets compared with the HP diet, indicating that providing a low-protein diet supplemented with rumen-protected AA led to greater N efficiency. There was no significant effect of treatment on plasma AA concentrations except for proline, which significantly increased for the MPLM treatment compared with the other 2 treatments. Overall, the results supported the concept that milk performance might be maintained when feeding lactating dairy cows with low CP diets if the absorbed AA balance is maintained through RPL and RPM feeding. Further investigations are needed to evaluate responses over a longer time period with consideration of all AA rather than on the more aggregated MP and the ratio between Lys and Met.

The effect of sex and dietary crude protein level on nutrient transporter gene expression and cecal microbiota populations in broiler chickens.

It is well known that male and female broilers differ in their growth performance and that many physiological factors contribute to this difference. The aim of this experiment is to investigate if there are differences between male and female broilers in cecal microbiota and nutrient transporter gene expression and if these differences play a role in the growth performance of broilers. The possible effect of protein level and its interaction with sex on microbiota and expression of the nutrient transporters were also investigated. Samples were collected from male and female birds fed either standard crude protein (SCP) or reduced crude protein diets (RCP) at the age of d 35. The experiment was designed as a 2 × 2 factorial arrangement of treatments consisting of 448 Cobb 500 broilers assigned to 32-floor pens with 4 treatments, 8 replicates, and 14 birds per pen for performance measurements. The factors were sex (male or female) and dietary crude protein (CP) level (standard or reduced). Body weight gain (BWG), feed intake and feed conversion ratio were recorded for each pen. Sex had a significant effect on BWG and FCR (P < 0.001) where males had a significantly higher BWG and better FCR compared to females. There was a significant interaction between sex and protein level on feed intake (FI) (P < 0.05), where male birds had a higher FI compared to female birds only when the birds were fed SCP but not RCP diets. There was a significant interaction between CP level and sex on the expression of CAT2 (P = 0.02) and PEPT2 (P = 0.026) where the genes were significantly upregulated in females but only when the RCP diet was fed. The RCP diet upregulated the expression of BoAT (P = 0.03) as a main effect. Female birds had significantly higher expression of the PepT-2 gene compared to the males. The alpha diversity of the cecal microbiota showed differences among the treatments. The Shannon diversity index was statistically higher (P = 0.036) for males fed the SCP diet and the Chao1 index for evenness was statistically higher (P = 0.027) in females fed the SCP diet. There was also a difference in the relative abundance of the 15 most common genera found in the cecal content of the broilers in this experiment and lastly, the differential composition of microbiota between the different treatments was also significantly different. This study suggests that chickens are able to compensate for a reduction in AA substrates when fed a low CP diet through the upregulation of certain AA transporters, females may adapt to low CP diets better by such upregulation compared to males, and lastly, sex has an effect on the cecal microbial population and these differences contribute towards the performance differences between male and female broilers.

Dietary strategies can increase cloacal endotoxin levels and modulate the resident microbiota in broiler chickens.

Endotoxins released from poultry feces have been associated with impaired human health. Because endotoxins are released from gram-negative intestinal bacteria, it was hypothesized that dietary strategies may influence endotoxin excretion via modulation of gut microbiota. We therefore tested dietary strategies that could potentially reduce cloacal endotoxin levels in broiler chickens. One-day-old male Ross 308 (N = 1,344) broilers were housed in 48 pens (N = 8 pens/treatment, 28 chickens per pen) and fed 1 of 6 diets for 35 days (d) in a 3-phase feeding program: a basic diet (CON) that served as the reference diet, or basic diet supplemented with butyrate (BUT), inulin (INU), medium-chain fatty acids (MCFA) or Original XPC™LS (XPC), or a high-fiber-low-protein (HF-LP) diet. A significant (P < 0.05) increase in cloacal endotoxin concentration at d 35 was observed in BUT as compared to CON. Analysis of cloacal microbiota showed a trend (P < 0.07) for a higher gram-negative/gram-positive ratio and for a higher relative abundance of gram-negative bacteria at d 35 (P ≤ 0.08) in BUT and HF-LP as compared to CON. A significant (P < 0.05) increase in average daily gain (ADG) and improved feed conversion ratio (P < 0.05) were observed in MCFA during the grower phase (d 14-28), and a significant (P < 0.05) increase in average daily feed intake (ADFI) was observed in MCFA during d 0 to 28. Broilers fed HF-LP had a significantly (P < 0.05) higher FCR and lower ADG throughout the rearing period. No treatment effects were found on footpad dermatitis, but BUT had worst hock burn scores at d 35 (P < 0.01) and MCFA had worst cleanliness scores at d 21 but not at d 35 (treatment*age P < 0.05), while INU had better cleanliness as compared to CON at d 35 (P < 0.05). In conclusion, especially BUT and HF-LP were able to modulate resident microbiota and BUT also increased cloacal endotoxin levels, which was opposite to our hypothesis. The present study indicates that cloacal endotoxin release can be affected by the diet but further study is needed to find dietary treatments that can reduce cloacal endotoxin release.

Digestibility and nitrogen balance of goats on high and low protein rations supplemented with a commercial tannin feed-additive.

Despite the increasing importance of goat production in response to high demand for their products and their relative robustness to environmental stressors, and in contrast to other ruminant species, little data is available on how tannin extract feeding affects their feed intake, nutrient digestion and nitrogen (N) metabolism. Therefore, a trial in Oman investigated the respective variables by using a commercial tannin feed additive. In a 4 (treatments) x 3 (periods) x 2 (animals) Youden square, two weaned Batinah bucks each were fed a high or low protein diet of Rhodes grass hay and crushed barley grain, with or without the addition of a chestnut and quebracho tannin extract at 2 g/kg metabolic weight. Feed offered, feed refused and faeces and urine excreted were quantified to determine diet digestibility, total N excretion, N retention and rumen microbial protein synthesis (MPS). Due to their young age and low live weight, feed intake of goats was relatively low. Crude protein level and tannin addition had no statistically significant effect on dry matter (DM) and N intake, DM digestibility, N excretion in faeces and urine, as well as MPS. In consequence, no benefit of tannin feeding could be confirmed for the goats' N retention, irrespective of diet composition. These results indicate, on one hand, an effective neutralisation of the tested tannin extract along the gastrointestinal tract of goats, but on the other hand, that stimulation of MPS or N retention by tannins cannot be evidenced when diet components are present that simultaneously release energy and protein, as is the case with barley.

Effect of antibiotics and low-crude protein diets on growth performance, health, immune response, and fecal microbiota of growing pigs.

This study aimed to investigate the effects of diets with and without antibiotics supplementation and diets with 18.5% and 13.0% crude protein (CP) on growth performance, carcass characteristics, disease incidence, fecal microbiota, immune response, and antioxidant capacity of growing pigs. One hundred and eighty pigs (59-day-old; 18.5 ±â€…2.5 kg) were distributed in a randomized complete block design in a 2 × 2 factorial arrangement, nine replicates, and five pigs per pen. The factors were CP (18.5% or 13.0%) and antibiotics (none or 100 mg/kg tiamulin + 506 mg/kg oxytetracycline). Medicated diets were fed from days 59 to 73. After that, all pigs were fed their respective CP diets from 73 to 87 days. Data were analyzed using the Mixed procedure in SAS version 9.4. From days 59 to 73, pigs fed antibiotics diets had higher (P < 0.05) average daily feed intake (ADFI), average daily weight gain (ADG), gain to feed ratio (G:F), compared to the diets without antibiotics. From days 73 to 87 (postmedicated period), any previous supplementation of antibiotics did not affect pig growth performance. Overall (days 59 to 87), pigs-fed antibiotics diets had higher (P < 0.05) G:F compared to pigs-fed diets without antibiotics. In all periods evaluated, pigs fed 18.5% CP diets had higher (P < 0.05) ADG and G:F compared to pigs fed 13.0% CP. Pigs fed the 13.0% CP diets had lower (P < 0.05) fecal score and diarrhea incidence than those fed 18.5% CP. Pigs fed 18.5% CP diets had improved (P < 0.05) loin area compared to pigs-fed diets with 13.0% CP. At 66 days of age, pigs-fed antibiotics diets had lower (P < 0.05) alpha diversity estimated with Shannon and Simpson compared to the pig-fed diets without antibiotics. At family level, pigs fed 18.5% CP diets had higher (P < 0.05) relative abundance of Streptococcaceae, and lower (P < 0.05) relative abundance of Clostridiaceae at days 66 and 87 compared with pigs fed 13.0% CP. Pigs-fed antibiotics diets had lower (P < 0.05) immunoglobulin G and protein carbonyl concentrations at day 66 compared to the pigs-fed diets without antibiotics. The reduction of dietary CP from 18.5% to 13.0% reduced the growth performance and loin muscle area of growing pigs, although it was effective to reduce diarrhea incidence. Antibiotics improved growth performance, lowered diarrhea incidence, improved components of the humoral immune response, and reduced microbiota diversity. However, in the postmedicated period, we found no residual effect on the general health of the animals, and considering the overall period, only G:F was improved by the use of antibiotics.

Dietary antibiotics have been used in pig farming practices to avoid health problems and improve animal growth performance. However, their use in production animals is considered a global health challenge, due to its association with selection of resistance in zoonotic bacteria. Another negative impact of pig farming that has gained attention is related to environmental pollution due to the excretion of nitrogenous compounds. Reducing dietary crude protein content has become a goal in the pig feed industry due to the limited availability and high cost of dietary protein sources, as well as the aim of enhancing gut health in pigs. Thus, the aim of this study was to investigate the effects of diets with and without antibiotics supplementation and diets with 18.5% and 13.0% crude protein for pigs. The reduction of dietary crude protein in this study reduced growth performance, although it was effective to reduce diarrhea incidence. Antibiotics improved growth performance, positively affected the overall health of animals, and reduced microbiota diversity. However, during the postmedicated period, we found no residual effect on the general health of the animals, and considering the overall period, only gain to feed ratio was improved by the use of antibiotics.

Effects of dietary protein content and crystalline amino acid supplementation patterns in low protein diets on intestinal bacteria and their metabolites in weaned pigs raised under Different sanitary conditions.

The objective of this experiment was to investigate the effects of dietary crude protein (CP) content and crystalline amino acids (CAA) supplementation patterns in low CP (LCP) diets on intestinal bacteria and their metabolites in weaned pigs raised under clean (CSC) or unclean sanitary conditions (USC). One hundred forty-four piglets (6.35 ±â€…0.63 kg) were assigned to one of six treatments in a 3 × 2 factorial arrangement based on CP content and sanitary conditions in a randomized complete block design to give eight replicates with three pigs per pen over a 21-d period. Diets consisted of a high CP (HCP; 21%) and two LCP (18%) diets supplemented with 9 CAA (Lys, Met, Thr, Trp, Val, Ile, Leu, His, and Phe) or only six CAA (Lys, Met, Thr, Trp, Val, and Ile) to meet the requirements. The CSC room was washed weekly, whereas the USC room had sow manure spread in the pens from the beginning of the study and was not washed throughout the experiment. Jejunum and colon digesta were sampled on day 21. Both jejunum and colon digesta were analyzed for ammonia nitrogen, short-chain fatty acids, and biogenic amines but only colon digesta was analyzed for microbiome composition (16s rRNA sequencing on MiSeq). Data were analyzed using R software for 16S rRNA and the MIXED procedure of SAS for microbial metabolites. Sanitation, CP content, and CAA supplementation patterns did not affect the diversity of colonic bacterial composition in weaned pigs. Pigs raised under USC had greater (P < 0.05) jejunal ammonia nitrogen concentration than those raised under CSC. Pigs fed LCP diets had reduced (P < 0.05) jejunal ammonia nitrogen concentration compared to those fed the HCP diet. Interactions between sanitation and dietary CP content were observed (P < 0.05) for: (1) jejunal acetate and (2) colonic spermidine and spermine, whereby (1) acetate concentrations decreased from NCP to LCP in pigs raised under the CSC but those concentrations increased under the USC, and (2) spermidine and spermine concentrations increased in LCP diets compared to HCP diet under USC, unlike CSC which did not show any difference between HCP and LCP. In conclusion, reducing dietary CP lowered ammonia nitrogen content regardless of sanitation and increased microbial metabolites in weaned pigs raised under USC. However, LCP diets with different CAA supplementation patterns did not affect bacterial diversity in weaned pigs, regardless of the hygienic conditions where the animals were housed.

Lowering dietary crude protein concentration by 3% to 4% units has been used as one of the strategies to promote growth and improve the gut health of weaned pigs. Undigested and endogenous protein could be available for microbial fermentation, and protein fermentation is considered detrimental to the gut health of the host animal. The unclean sanitary condition model mimics commercial raising conditions and stimulates a low-grade inflammatory and immune response. Ammonia nitrogen is one of the harmful metabolites derived by protein fermentation and pigs fed low-protein diets had decreased ammonia nitrogen than those fed high-protein diets. Also, pigs raised under unclean sanitation had greater ammonia nitrogen than those raised under clean sanitation. However, sanitation, protein content, and crystalline amino acids supplementation patterns did not affect the diversity of colonic bacterial composition in weaned pigs. The results obtained from the present study showed that a low protein diet could be used to improve gut health in weaned pigs.

Efficacy of oat hulls varying in particle size in mitigating performance deterioration in broilers fed low-density crude protein diets.

Dietary fibres with increased particle size may develop foregut more efficiently in meat poultry fed diets moderately low in crude protein. The study investigated the performance of broilers fed low-density crude protein diets with the inclusion of oat hulls increasing in particle size. Ross 308 male broiler chicks (n = 336) were divided into 48 pens, 12 pens per treatment (7 chicks per pen). Pens were allocated to 4 dietary treatments in mash form; positive control (PC), a standard crude protein diet, negative control (NC), around 5% lower in protein with 5% celite as an inert material, OH400: 5% lower protein diet with 5% oat hulls of geometric mean diameter (GMD) 400 µm, OH850: 5% lower protein diet with 5% oat hulls of GMD 850µm. Birds were fed ad libitum from d 1 to 35 in 3 phases; starter: d 0 to 10, grower: d 10 to 24 and finisher: d 24 to 35. Growth performance was calculated at the end of the trial. Two birds per pen were sampled on d 24 and 35 to collect data on proventriculus and gizzard weights and pooled ileal digesta. Apparent ileal digestibility of amino acids on d 24, and metabolizable energy on d 24 and 35 were recorded for each pen. Litter was sampled from each pen on d 34 to analyze litter N and moisture content. Footpad dermatitis scores of all birds per pen were recorded on d 35. Overall, no difference (P > 0.05) in body weight gain and feed intake was found between the treatments. However, NC and OH400 showed poorer FCR than PC, whereas FCR on PC and OH850 was similar (P > 0.05). Gizzard absolute weight and relative to body weight, and gizzard to proventriculus ratio were higher (P < 0.05) on OH850 compared to PC and NC on d 24 and 35. Gizzard digesta particle size was reduced (P < 0.05) on OH850 compared to all other diets on d 24 and 35. Amino acids digestibility coefficients for aspartic acid and valine increased (P < 0.05) in birds fed OH850 compared to PC, whereas coefficients for several other amino acids were improved compared to NC and OH400. The apparent ileal digestibility of metabolizable energy was similar (P > 0.05) between PC and OH850. Litter moisture and nitrogen, and footpad dermatitis scores were reduced (P < 0.05) on OH850 compared to PC. In conclusion, the inclusion of coarse oat hulls of GMD 850 µm in low-density crude protein diets can be beneficial for the broilers in developing the foregut, utilizing the nutrients efficiently and reducing litter nitrogen and moisture, and footpad scores.

Effects of a low-protein diet supplemented with essential amino acids on egg production performance and environmental gas emissions from layer-manure composting in laying hens in the later laying period.

We fed 330-545 day-old laying hens (later laying period) a low-protein diet supplemented with essential amino acids (LPS) and composted their manure. We then investigated the laying performance of the hens, the nitrogen balance and emission of nitrous oxide (N2 O), methane (CH4 ), and ammonia (NH3 ) from the composting, and several characteristics of the finished compost. There were no significant differences in the egg-laying rate, egg mass, egg weight, proximate compositions in egg yolk and egg white, or feed intake between the laying hens fed a Control diet (Cont) and those fed the LPS diet. However, the LPS-fed hens had lower excreta levels and nitrogen excretion. In addition, the environmental gas emissions per layer from composting of the manure from the LPS-fed laying hens were decreased by 9.7% for N2 O, 40.9% for CH4 , and 24.8% for NH3 compared to the Cont-fed laying hens. The concentrations of total nitrogen in the finished compost were similar between the LPS-fed and Cont-fed laying hens. In a vegetable-growth test, the weights of komatsuna plants grown with compost from LPS-fed and Cont-fed hens were also not significantly different. Feeding an LPS diet to 330-545 day-old laying hens was suggested to reduce environmental gas emissions from manure composting without affecting the egg production performance.

Effects of dietary niacinamide and CP concentrations on the nitrogen excretion, growth performance, and meat quality of pigs.

Reducing the dietary CP concentration in the formulation of low-protein diets without adverse effects on animal growth performance and meat quality remains challenging. In this study, we investigated the effects of nicotinamide (NAM) on the nitrogen excretion, growth performance, and meat quality of growing-finishing pigs fed low-protein diets. To measure the nitrogen balance, we conducted two trials: in nitrogen balance trial 1, four crossbred (Duroc × Landrace × Large White) barrows (40 ± 0.5 kg BW) were used in a 4 × 4 Latin square design with four diets and periods. The diets consisted of a basal diet + 30 mg/kg NAM (a control dose), basal diet + 90 mg/kg NAM, basal diet + 210 mg/kg NAM, and basal diet + 360 mg/kg NAM. In nitrogen balance trial 2, another four barrows (40 ± 0.5 kg BW) were used in a 4 × 4 Latin square design. The diets consisted of a basal diet + including 30 mg/kg NAM (control), basal diet + 360 mg/kg NAM, low-protein diet + 30 mg/kg NAM, and low-protein diet + 360 mg/kg NAM. To measure growth performance, two trials were conducted. In growth performance trial 1, 40 barrows (37.0 ± 1.0 kg) were randomly allocated to one of four dietary treatments (n = 10 per group), whereas in growth performance trial 2, 300 barrows (41.4 ± 2.0 kg) were randomly allocated to one of four dietary treatments, with each dietary treatment conducted in five repetitions with 15 pigs each. The four diets in the two growth performance trials were similar to those in nitrogen balance trial 2. Supplementing the diet with 210 or 360 mg/kg NAM reduced urinary nitrogen excretion and total nitrogen excretion and increased nitrogen retention comparted with the control diet (P < 0.05). Compared with the control diet, the low-protein diet with 360 mg/kg NAM reduced faecal, urinary, and total nitrogen excretion (P < 0.05) without affecting nitrogen retention and average daily gain (P > 0.05). Pigs fed the low-protein diet with 360 mg/kg NAM showed a decreased intramuscular fat content in the longissimus thoracis muscle when compared with pigs fed the control diet (P > 0.05). Our results suggest NAM as a suitable dietary additive to reduce dietary CP concentration, maximise nitrogen retention and growth performance, and decrease fat deposition in pigs.

Protective effects of genistein on the production performance and lipid metabolism disorders in laying hens with fatty liver hemorrhagic syndrome by activation of the GPER-AMPK signaling pathways.

The aim of this study was to evaluate the beneficial effects and potential mechanisms of genistein (GEN) on production performance impairments and lipid metabolism disorders in laying hens fed a high-energy and low-protein (HELP) diet. A total of 120 Hy-line Brown laying hens were fed with the standard diet and HELP diet supplemented with 0, 50, 100, and 200 mg/kg GEN for 80 d. The results showed that the declines in laying rate (P < 0.01), average egg weight (P < 0.01), and egg yield (P < 0.01), and the increase of the ratio of feed to egg (P < 0.01) induced by HELP diet were markedly improved by 100 and 200 mg/kg of GEN treatment in laying hens (P < 0.05). Moreover, the hepatic steatosis and increases of lipid contents (P < 0.01) in serum and liver caused by HELP diet were significantly alleviated by treatment with 100 and 200 mg/kg of GEN in laying hens (P < 0.05). The liver index and abdominal fat index of laying hens in the HELP group were higher than subjects in the control group (P < 0.01), which were evidently attenuated by dietary 50 to 200 mg/kg of GEN supplementation (P < 0.05). Dietary 100 and 200 mg/kg of GEN supplementation significantly reduced the upregulations of genes related to fatty acid transport and synthesis (P < 0.01) but enhanced the downregulations of genes associated with fatty acid oxidation (P < 0.01) caused by HELP in the liver of laying hens (P < 0.05). Importantly, 100 and 200 mg/kg of GEN supplementation markedly increased G protein-coupled estrogen receptor (GPER) mRNA and protein expression levels and activated the AMP-activated protein kinase (AMPK) signaling pathway in the liver of laying hens fed a HELP diet (P < 0.05). These data indicated that the protective effects of GEN against the decline of production performance and lipid metabolism disorders caused by HELP diet in laying hens may be related to the activation of the GPER-AMPK signaling pathways. These data not only provide compelling evidence for the protective effect of GEN against fatty liver hemorrhagic syndrome in laying hens but also provide the theoretical basis for GEN as an additive to alleviate metabolic disorders in poultry.

Fatty liver hemorrhagic syndrome (FLHS) is a nutritional and metabolic disease that seriously threatens the health and performance of laying hens, which is characterized by hepatic steatosis and lipid metabolism disorders. As an isoflavone phytoestrogen, genistein (GEN) exerts many beneficial functions, including alleviating lipid metabolism disorders and anti-inflammatory properties. However, further research is needed on the protective effect and potential mechanism of GEN on the FLHS in laying hens. Here, we found that GEN treatment improved liver injury and decline of production performance in laying hens with FLHS. Moreover, GEN treatment alleviated hepatic steatosis and lipid metabolism disorders through reducing the expression levels of mRNA related to fatty acid transport and synthesis and enhancing the mRNA expression levels of factors associated with fatty acid oxidation in FLHS layers, which may be achieved by activation of the G protein-coupled estrogen receptor­adenosine 5'-monophosphate (AMP)-activated protein kinase signaling pathways. These data not only provide compelling evidence for the protective effects and mechanisms of GEN against FLHS in laying hens but also provide the theoretical basis for GEN to alleviate other metabolic disorders in poultry.

The effects of protease, xylanase, and xylo-oligosaccharides on growth performance, nutrient utilization, short-chain fatty acids, and microbiota in Eimeria-challenged broiler chickens fed low-protein diet.

A total of 392 Cobb 500 off-sex male broiler chicks were used in a 21-day experiment to study the effect of protease, xylanase, and xylo-oligosaccharides (XOS) on improving growth performance, nutrient utilization (ileal digestibility and total tract retention), gene expression of nutrient transporters, cecal short-chain fatty acids (SCFAs), and microbiota profile of broilers challenged with Eimeria spp. Chicks at 0-day old were allocated to 8 treatments in a 4 × 2 factorial arrangement: 1) corn-soybean meal diet with no enzyme (Con); 2) Con plus 0.2 g/kg protease alone (PRO); 3) Con plus 0.2 g/kg protease combined with 0.1 g/kg xylanase (PRO + XYL); or 4) Con plus 0.5 g/kg xylo-oligosaccharides (XOS); with or without Eimeria challenge. The 4 diets were formulated to be marginally low in crude protein (183 g/kg). Challenged groups were inoculated with a solution containing E. maxima, E. acervulina, and E. tenella oocysts on d 15. Eimeria depressed (P < 0.01) growth performance and nutrient utilization. Supplemental protease improved (P < 0.05) body weight gain and feed intake in the prechallenge phase (d 0-15) but had no effect during the infection period (d 15-21). There was no interaction between infection and feed supplementation for nutrient utilization. The supplementations of either PRO or XOS alone increased (P < 0.01) total tract retention of Ca and tended (P < 0.1) to improve total tract retention of N, P, AME, and AMEn. Eimeria decreased (P < 0.05) expressions of GLUT2, GLUT5, PepT1, ATP2B1, CaSR, Calbidin D28K, NPT2, and ZnT1 but increased (P < 0.01) expression of GLUT1. XOS supplementation increased (P < 0.05) ATP2B1 expression. Protease decreased (P < 0.05) isobutyrate concentration in unchallenged treatments but not in challenged treatments. Eimeria decreased (P < 0.01) cecal saccharolytic SCFAs acetate and propionate but increased (P < 0.01) branched-chain fatty acid isovalerate. The supplementation of PRO + XYL or XOS increased (P < 0.05) cecal butyrate or decreased cecal isobutyrate concentrations, respectively. PRO + XYL and XOS decreased cecal protein levels in unchallenged birds but not challenged ones. Eimeria challenge significantly (P < 0.05) decreased the microbial richness (Observed features) and diversity (Shannon index and phylogenetic diversity) and changed the microbial composition by reducing the abundance of certain bacteria, such as Ruminococcus torques, and increasing the abundance of others, such as Anaerostipes. In contrast, none of the additives had any significant effect on the cecal microbial composition. In conclusion, PRO or XOS supplementation individually improved nutrient utilization. All the additives decreased the cecal content of branched-chain fatty acids, consistent with decreased cecal N concentration, although the effects were more pronounced in unchallenged birds. In addition, none of the feed additives impacted the Eimeria-induced microbial perturbation.

The effects of arginine and branched-chain amino acid supplementation to reduced-protein diet on intestinal health, cecal short-chain fatty acid profiles, and immune response in broiler chickens challenged with Eimeria spp.

We investigated the effects of supplementing arginine (Arg) and branched-chain amino acids (BCAA) in broilers fed reduced-protein diets and challenged with Eimeria spp. All birds were fed the same starter diet meeting Cobb 500 nutrient specifications from d 1 to 9. Four grower diets: positive control (PC) with 20.0% crude protein (CP); reduced-protein negative control (NC) with 17.5% CP; or NC supplemented with Arg or BCAA at 50% above recommendations (ARG or BCAA) were fed to the birds from d 9 to 28. Birds were allocated in a 2 × 4 factorial arrangement (4 diets, each with or without challenge), with 8 replicates per treatment. On d 14, the challenge groups were orally gavaged with mixed Eimeria spp. Intestinal permeability was higher (P < 0.05) in NC than PC, whereas the permeability of ARG and BCAA groups did not differ significantly from PC. On d 28, a significant interaction (P < 0.01) was observed in CD8+: CD4+ ratios in cecal tonsils (CT), Eimeria challenge increased the ratios in all groups except for the ARG group. On d 21, a significant interaction was found for CD4+CD25+ percentages in CT (P < 0.01) that Eimeria challenge increased the percentages only in PC and NC groups. On d 21 and 28, significant interactions (P < 0.01) were found for macrophage nitric oxide (NO) production. In nonchallenged birds, NO was higher in the ARG group than other groups, but in challenged birds, NO was higher in both ARG and BCAA groups. On d 21, a significant interaction was found for bile anticoccidial IgA concentrations (P < 0.05) that Eimeria challenge increased IgA only in NC and ARG groups. The results suggest that a reduced-protein diet exacerbates the impact of the Eimeria challenge on intestinal integrity, but this could be mitigated by Arg and BCAA supplementations. Arginine and BCAA supplementations in reduced-protein diets could be beneficial for broilers against Eimeria infection by enhancing the immune responses. The beneficial effects of Arg supplementation tended to be more pronounced compared to BCAA supplementation.

Effect of exogenus protease on performance, nutrient digestibility, intestinal histomorphometric, meat quality characteristics, carcass yield in broilers fed low protein diets.

The objective of the present study was to evaluate the effects of increasing doses of protease on broilers from 1 to 42 days of age. A total of 1290 Ross AP broilers were used, distributed among five treatments: positive control diet, negative control diet (NC), NC + 50 ppm of protease, NC + 100 ppm of protease, and NC + 200 ppm of protease. Each treatment contained six replicates of 43 animals each. The inclusion of proteases in the diet had effects (P < 0.05) on body weight, feed intake, weight gain, and feed conversion in the 12 to 21 day period; body weight, weight gain, and feed intake in the 29 to 42 day period; nutrient digestibility (energy metabolizability coefficient and crude protein at 28 days); and intestinal parameters (crypt and muscle width of jejunum and ileum at 28 days and villus length, crypt length, and jejunum thickness muscle layer at 42 days). These results indicate that the inclusion of protease in broiler feed can improve production parameters when the amount of crude protein in the diet is reduced.

Effect of supplementing rumen-protected methionine, lysine, and histidine to low-protein diets on the performance and nitrogen balance of dairy cows.

Lowering the dietary protein content can reduce N excretions and NH3 emissions from manure and increase milk N efficiency of dairy cows. However, milk yield (MY) and composition can be compromised due to AA deficiency. Methionine and Lys are known as first limiting EAA for dairy cows, and recently His is also mentioned as limiting, especially in grass-based or low-protein diets. To examine this, a trial was conducted with a 3-wk pre-experimental adaptation period (diet 16.5% crude protein), followed by a depletion period of 4 wk, in which 39 cows (average ± standard deviation: 116 ± 29.3 d in milk, 1.8 ± 1.2 lactations, 638 ± 73.2 kg of body weight, and 32.7 ± 5.75 kg MY/d) received a low-protein diet (CTRL) (14.5% crude protein). Then, taking into account parity, His plasma concentration, and MY, cows were randomly assigned to 1 of 3 treatment groups during the rumen-protected (RP) AA period of 7 wk; (1) CTRL; (2) CTRL + RP-Met + RP-Lys (MetLys); (3) CTRL + RP-Met + RP-Lys + RP-His (MetLysHis). Products were dosed, assuming requirements for digestible (d) Met, dLys, and dHis being, respectively, 2.4%, 7.0%, and 2.4% of intestinal digestible protein. In the cross-back period of 5 wk, all cows received the CTRL diet. During the last week of each period, a N balance was conducted by collecting total urine and spot samples of feces. Total feces production was calculated using the inert marker TiO2. Statistical analysis was performed with a linear mixed model with cow as random effect and data of the last week of the pre-experimental period used as covariate for the animal performance variables. No effect of supplementing RP-Met and RP-Lys nor RP-Met, RP-Lys, and RP-His on feed intake, milk performance, or milk N efficiency was observed. However, the plasma AA profile indicated additional supply of dMet, dLys, and dHis. Nevertheless, evaluation of the AA uptake relative to the cow's requirements showed that most EAA (exclusive Arg and Thr) were limiting over the whole experiment. Only dHis was sufficiently supplemented during the RP-AA period due to an overestimation of the diet's dMet and dLys supply in the beginning of the trial. The numerically increased milk urea N and urinary N excretion when RP-Met, RP-Lys, and RP-His were added to the low-protein diet suggest an increased catabolism of the excess His.

Low-protein diets supplemented with isoleucine alleviate lipid deposition in broilers through activating 5' adenosine monophosphate-activated protein kinase and janus kinase 2/signal transducer and activator of transcription 3 signaling pathways.

This study aimed to evaluate the effect of isoleucine (Ile) on growth performance, meat quality and lipid metabolism of broilers fed a low-protein diet (LPD). The 396 one-day-old male Cobb broilers were allocated to 4 treatment groups as follows: control diet (CON), LPD, LPD + 0.13% Ile (LPD-LI) and LPD + 0.26% Ile (LPD-HI), with nine replicates of 11 broilers each for 42 d. The Ile increased average daily gain, average daily feed intake, fiber density and the mRNA level of myosin heavy chain (MyHC)-I in breast muscle, and decreased feed to gain ratio, shear force, fiber diameter and the mRNA level of MyHC-IIb in breast muscle, which were impaired by the LPD. Compared to the LPD group, broilers in LPD-LI and LPD-HI groups had lower serum lipid levels, liver fat content, abdominal adipose percentage and mRNA levels of peroxisome proliferator-activated receptor-γ, CCAAT/enhancer binding protein-α, ki-67, topoisomerase II alpha (TOP2A) and thioredoxin-dependent peroxidase 2 in abdominal adipose and liver X receptors-α, sterol regulatory element binding protein 1 (SREBP1), acetyl-CoA carboxylase (ACC) and fatty acid synthase (FAS) in liver, and higher mRNA levels of peroxisome proliferator activated receptor-α, carnitine palmitoyl-transferase 1 (CPT-1), and acyl-CoA oxidase 1 (ACOX1) in liver, which were equal to the CON levels. A LPD supplemented with Ile decreased enzyme activities of ACC and FAS in liver and glycerol-3-phosphate dehydrogenase and TOP2A in abdominal adipose, and increased enzyme activities of CPT-1 and ACOX1 in liver. Furthermore, Ile supplementation enhanced the mRNA level of leptin receptor and protein levels of phospho-5' adenosine monophosphate-activated protein kinase (AMPK), mechanistic target of rapamycin, ribosomal protein 70 S6 kinase, janus kinase 2 (JAK2), and signal transducer and activator of transcription 3 (STAT3), and decreased the protein level of SREBP1 in the liver of broilers in LPD group. In conclusion, dietary supplementation with Ile to 0.83% could improve growth performance and meat quality and alleviate lipid deposition of broilers fed a LPD through activating AMPK and JAK2/STAT3 signaling pathways.

Different amino acid supplementation patterns in low-protein diets on growth performance and nitrogen metabolism of goslings from 1 to 28 days of age.

The investigation aimed to explore the suitable amino acid (AA) supplementation pattern for goslings under low-protein diets. A total of 364 1-day-old male goslings were randomly divided into 4 experimental groups, with 7 pens containing 13 goslings each. The 4 groups were control (CP, 18.55%), LPM (CP, 15.55% + major AA), LPA (CP, 15.55% + all AA), and LPR (CP, 15.55% + AA content reduced proportionally to the control's CP). The corn-soybean meal diets are formulated according to the ideal AA model of goose and its nutritional requirements. The results indicated that the ADG and BW were the lowest, and the F: G was the highest in LPR (P < 0.05); the other three groups were not significantly different (P > 0.05). The ADFI and mortality were not different among all the groups (P > 0.05). Among the AA content in serum and breast muscle, lysine in serum significantly decreased compared with the control (P < 0.05). The UREA content was approximately 2-fold higher in the LPR group than in the LPM and LPA groups (P < 0.05). No difference in IgA, IgG, IgM, and IgE levels was observed among the groups (P > 0.05). The nitrogen excretion was decreased in LPM and LPA compared to the control and LPR (P < 0.05). Nitrogen deposition did not differ among groups (P > 0.05). Nitrogen utilization was highest in the LPA and LPM groups, followed by the control group and LPR (P < 0.05). In conclusion, the patterns of supplementation of major AA and all AA in low-protein diets (CP, 15.55%) had no adverse effect on the growth performance compared with the control (CP, 18.55%) of the goslings. Besides, the two patterns could decrease nitrogen excretion and increase nitrogen utilization. Furthermore, from the perspective of dietary cost and environmental protection, the pattern of supplementing major AA in a corn-soybean meal low-protein diet is suggested.

Effects of lowering dietary protein content without or with increased protein-bound and feed-grade amino acids supply on growth performance, body composition, metabolism, and acute-phase protein of finishing pigs under daily cyclic heat stress.

This study investigated the effects of a low-protein diet with or without an increase in dietary protein and feed-grade amino acids (AAs) on the growth performance, body composition, metabolism, and serum acute-phase proteins of finishing pigs reared in thermoneutrality or cyclic heat stress conditions. A total of 90 gilts (67.7 ± 6.2 kg) were distributed in a 2 × 3 factorial arrangement (two ambient temperatures and three diets). Ambient temperatures (AT) were thermoneutral (TN, 22 °C for 24 h) and cyclic heat stress (CHS, 12 h to 35 °C and 12 h to 22 °C). The evaluated diets (D) were high crude protein (HP); low CP-free AA-supplemented diets (LPAAs); low CP-free AA-supplemented diets and digestible Lys level (+20%), and Lys:AA ratios above recommendations (LPAA+). The experimental period lasted 48 d (two experimental phases: days 0-27 and days 28-48, respectively). CHS pigs had higher skin temperature (P < 0.05) than TN pigs. Pigs in CHS had higher rectal temperature (P < 0.05) than TN pigs until day 38 but similar (P > 0.10) to TN pigs from 38 to 45 d. For the entire experiment, CHS pigs had lower (P < 0.05) final BW, average daily gain and daily feed intake, net energy intake, body lipid, bone mineral, lipid deposition, energy retention, Lys and CP intake, and nitrogen excretion than TN pigs. The level of CP intake impacted nitrogen excretion, nitrogen retention efficiency, and urea as pigs fed HP had the highest values, and pigs fed LPAA had the lowest values (P < 0.05). On day 27, CHS pigs had lower (P < 0.05) free triiodothyronine than TN pigs. LPAA+ pigs had lower (P < 0.05) insulin than LPAA. On day 48, CHS pigs had lower (P < 0.05) thyroxine, albumin, and lactate than TN pigs. On day 27, pigs fed LPAA+ had higher (P < 0.05) lactate than pigs fed HP or LPAA. Both AT and D were enough to stimulate the immune system as CHS pigs had lower (P < 0.05) transferrin and 23-kDa protein levels than TN pigs, and HP pigs had higher haptoglobin than LPAA on day 27. These results confirm the deleterious effects of high AT on performance, body composition, metabolism, and immune system stimulation in finishing pigs. These data also show that a diet with low levels of CP can be provided to pigs in CHS without affecting performance and body composition while reducing nitrogen excretion. However, the use of a diet with an AA level above the requirements obtained by increasing intact protein and free AA did not attenuate the impact of CHS on performance and body composition of pigs.

High ambient temperature and air humidity are the most important climatic factors that jeopardize pig production. Multiple strategies can be applied for pigs under heat stress, including recent research to improve understanding the use of nutrition to attenuate the impact of heat stress. Heat stress impairs digestion, absorption, and amino acid metabolism with changes in amino acid requirements. Updates on the nutritional assessment strategies by differing the diets by protein and amino acid content (protein-bound or feed-grade) seem to be efficient tools for pork producers as amino acids play a functional role in challenged pigs apart from the beneficial effects on performance.

Limiting amino acids supplementation in low crude protein diets and their impacts on growth performance and carcass composition in Labeo rohita (rohu) adult fish / Limitação da suplementação de aminoácidos em dietas com baixo teor de proteína bruta e seus impactos no desempenho de crescimento e composição da carcaça em peixes adultos Labeo rohita (rohu)

Ninety days study was conducted in hapas installed in earthen ponds. Fish of an average initial weight (220g) were evenly distributed in triplicate groups within fifteen hapas. Five experimental diets labeled as T1 (25% CP and NRC recommended amino acid level) as control diet, T2 (with 2% low protein and 5% amino acid supplementation), T3 (with 2% low protein and 10% amino acid supplementation), T4 (with 4% low protein and 10% amino acid supplementation) and T5 (with 4% low protein and 20% amino acid supplementation) were prepared. Fish were fed with @3% of their body weight twice a day at 10.00 & 16:00 hour. Significantly higher percent weight gain (420.18 ± 66.84a) and specific growth rate (13499.33±1273.54a) along with improved feed conversion ratio (1.29 ± 0.09b) and hundred percent survivals were recorded during the trial. Furthermore proximate analysis of meat showed significant improvement in the crude protein level (81.77 ± 0.19a) served with diet containing 20% limiting amino acids mixture. Therefore, limiting amino acids can be a source of cost effective feed and use safely in L. rohita diet.

O estudo de 90 dias foi realizado em hapas instalados em tanques de terra. Peixes com peso inicial médio (220 g) foram distribuídos uniformemente em grupos triplicados em 15 hapas. Cinco dietas experimentais rotuladas como T1 (25% de CP e NRC recomendado nível de aminoácidos) como dieta controle, T2 (com 2% de proteína baixa e 5% de suplementação de aminoácidos), T3 (com 2% de proteína baixa e 10% de suplementação de aminoácidos), T4 (com 4% de baixa proteína e 10% de suplementação de aminoácidos) e T5 (com 4% de baixa proteína e 20% de suplementação de aminoácidos) foram preparadas. Os peixes foram alimentados com 3% do seu peso corporal duas vezes por dia às 10h00 e 16h00. Ganho de peso significativamente maior (420,18 ± 66,84a) e taxa de crescimento específico (13499,33 ± 1273,54a) juntamente com taxa de conversão alimentar melhorada (1, 29 ± 0,09b) e sobrevivência de cem por cento foram registrados durante o ensaio. Além disso, a análise aproximada da carne mostrou melhora significativa no nível de proteína bruta (81,77 ± 0,19a) servida com dieta contendo 20% de mistura de aminoácidos limitantes. Portanto, a limitação de aminoácidos pode ser uma fonte de alimentação econômica e usada com segurança na dieta de L. rohita.

Effect of composite enzyme supplementation on production parameters, intestinal segment measurements, and apparent nutrient digestibility of broiler chickens fed low energy and protein diets.

A 42-day study was conducted to consider the effect of composite enzyme (Natuzyme®) supplementation on production parameters, intestinal segment measurements, and nutrient digestibility of broiler chickens fed low energy and protein (LEP) diets. Two hundred male Ross 308 broiler chicks consisting of 4 groups were divided into five pens (10 chicks/pen). The first group (control) received a standard diet without composite enzyme (CE) supplementation, whereas the LEP0, LEP0.25, and LEP0.50 groups received LEP supplemented with CE at 0, 0.25, and 0.50 g/kg feed, respectively. Compared to birds in the control group, supplementation of 0.50 g/kg CE to LEP diet offset (P < 0.05) the poor feed intake (FI), body weight gain (BWG), feed conversion ratio (FCR), and cost/kg gain observed among LEP0 and LEP0.25 groups on days 0-21, 22-42, and 0-42. Birds in the LEP0, followed by the LEP0.25 group, had decreased (P < 0.05) ileum weight, villi height, villus height, and crypt depth ratio (VH/CD), and deeper crypt depth of duodenum and ileum compared to birds in the control group. However, the aforementioned parameters were improved in the LEP0.50 group. Likewise, supplementation of CE at 0.50 g/kg to broiler diet having LEP improved (P < 0.05) digestibility of energy, fiber, and protein on day 42. In conclusion, CE supplemented at 0.50 g/kg diet ameliorated the negative effect caused by feeding LEP diets on production parameters, ileum weight, villi height, VH/CD, and deeper crypt depth of duodenum and ileum as well as energy, fiber, and protein digestibility of broiler chickens.