UCHL1 promotes the proliferation of porcine granulosa cells by stabilizing CCNB1

Background The proliferation of porcine ovarian granulosa cells(GCs)is essential to follicular development and the ubiquitin–proteasome system is necessary for maintaining cell cycle homeostasis.Previous studies found that the deubiquitinase ubiquitin carboxyl-terminal hydrolase 1(UCHL1)regulates female reproduction,especially in ovarian development.However,the mechanism by which UCHL1 regulates porcine GC proliferation remains unclear.Results UCHL1 overexpression promoted GC proliferation,and knockdown had the opposite effect.UCHL1 is directly bound to cyclin B1(CCNB1),prolonging the half-life of CCNB1 and inhibiting its degradation,thereby promoting GC proliferation.What's more,a flavonoid compound-isovitexin improved the enzyme activity of UCHL1 and promoted the proliferation of porcine GCs.Conclusions UCHL1 promoted the proliferation of porcine GCs by stabilizing CCNB1,and isovitexin enhanced the enzyme activity of UCHL1.These findings reveal the role of UCHL1 and the potential of isovitexin in regulating proliferation and provide insights into identifying molecular markers and nutrients that affect follicle development.

Advances in low-protein diets for swine

Recent years have witnessed the great advantages of reducing dietary crude protein(CP) with free amino acids(AA)supplementation for sustainable swine industry, including saving protein ingredients, reducing nitrogen excretion, feed costs and the risk of gut disorders without impairing growth performance compared to traditional diets. However, a tendency toward increased fatness is a matter of concern when pigs are fed low-protein(LP) diets. In response, the use of the net energy system and balanced AA for formulation of LP diets has been proposed as a solution. Moreover, the extent to which dietary CP can be reduced is complicated. Meanwhile, the requirements for the first five limiting AA(lysine, threonine, sulfur-containing AA, tryptophan, and valine) that growing-finishing pigs fed LP diets were higher than pigs fed traditional diets, because the need for nitrogen for endogenous synthesis of non-essential AA to support protein synthesis may be increased when dietary CP is lowered. Overall, to address these concerns and give a better understanding of this nutritional strategy, this paper reviews recent advances in the study of LP diets for swine and provides some insights into future research directions.

Study and use of the probiotic Lactobacillus reuteri in pigs:a review

Probiotics are living microorganisms that provide a wide variety of health benefits to the host when ingested in adequate amounts. The bacterial strains most frequently used as probiotic agents are lactic acid bacteria, such as Loctobocillus reuteri, which is one of the few endogenous Lactobadllus species found in the gastrointestinal tract of vertebrates, including humans, rats, pigs and chickens. L. reuteri is one of the most well documented probiotic species and has been widely utilized as a probiotic in humans and animals for many years. Initially, L. reuteri was used in humans to reduce the incidence and the severity of diarrhea, prevent colic and necrotic enterocolitis, and maintain a functional mucosal barrier. As interest in alternatives to in-feed antibiotics has grown in recent years, some evidence has emerged that probiotics may promote growth, improve the efficiency of feed utilization, prevent diarrhea, and regulate the immune system in pigs. In this review, the characteristics of L. reuteri are described, in order to update the evidence on the efficacy of using L. reuteri in pigs.

Studies on Reducing Nitrogen Excretion: Ⅰ. Net Energy Requirement of Finishing Pigs Maximizing Performance and Carcass Quality Fed Low Crude Protein Diets Supplemented with Crystalline Amino Acids

Two experiments were conducted to in- vestigate the effects of net energy （NE） level on the performance and carcass traits of finishing pigs fed low crude protein （CP） diets supplemented with crystalline amino acids （CAA）. A total of 216 （Exp. 1） and 360 （Exp. 2 ） barrows （ Yorkshire × Landrace× Duroc ） were allotted to one of six treatments （n =6）. The experimental diets were based on corn and soybean meal, and consisted of a high-CP diet that contained approximately 16% CP and 2.50 Mcal/kg of NE as well as five low-CP diets in which the CP level of the diet was reduced by approximately four percentage units but was supplemented with crystalline lysine, methionine, threonine, and tryptophan. The low CP diets provided 2.64, 2.58, 2.50, 2.42, or 2.36 Mcal/kg in Exp. 1 as well as 2.45, 2.40, 2.35, 2.30, or 2.25 Mcai/kg in Exp.2. In Exp. 1, a linear （P=0.03） increase in weight gain was observed with decreasing NE level while the ratio of gain to feed was unaffect- ed （P 〉 0.05） by NE level. There was a linear in-crease （P = 0.01 ） in the percentage of fat-free lean and a linear decrease （ P = 0.03 ） in the percentage of total fat with decreasing NE levels. In Exp. 2, a sig- nificant quadratic （ P = 0.03 ） effect of NE level was observed for weight gain. The ratio of gain to feed demonstrated a significant （P 〈0.01 ） quadratic effect with pigs fed 2.35 and 2.40 Mcal/kg of NE, Pigs fed the diet containing 2.40 Mcal/kg of NE had the lowest percentage of total fat （36. 95% ） and the highest percentage of fat-free lean （49.36%）. The overall results of these experiments indicate that feed- ing either a surplus or a deficiency of NE is detrimen- tal to both pig performance and carcass composition when low CP diets supplemented with CAA are fed. Our results indicate that when the CP content of the diet is reduced by four percentage units and the diet is properly supplemented with CAA, maximum per- formance and carcass quality will be obtained if the diet provides approximately 2.42 Mcal/kg of NE.

Novel metabolic and physiological functions of branched chain amino acids: a review

It is widely known that branched chain amino acids（BCAA） are not only elementary components for building muscle tissue but also participate in increasing protein synthesis in animals and humans. BCAA（isoleucine, leucine and valine） regulate many key signaling pathways, the most classic of which is the activation of the m TOR signaling pathway. This signaling pathway connects many diverse physiological and metabolic roles. Recent years have witnessed many striking developments in determining the novel functions of BCAA including：（1） Insufficient or excessive levels of BCAA in the diet enhances lipolysis.（2） BCAA, especially isoleucine, play a major role in enhancing glucose consumption and utilization by up-regulating intestinal and muscular glucose transporters.（3）Supplementation of leucine in the diet enhances meat quality in finishing pigs.（4） BCAA are beneficial for mammary health, milk quality and embryo growth.（5） BCAA enhance intestinal development, intestinal amino acid transportation and mucin production.（6） BCAA participate in up-regulating innate and adaptive immune responses.In addition, abnormally elevated BCAA levels in the blood（decreased BCAA catabolism） are a good biomarker for the early detection of obesity, diabetes and other metabolic diseases. This review will provide some insights into these novel metabolic and physiological functions of BCAA.

Effects of L-lysine·H2SO4 product on the intestinal morphology and liver pathology using broiler model

Background: Lysine is used widely in livestock production due to the shortage of feed protein resources.Llysine·H2SO4 contains L-lysine sulphate as well as fermentation co-products which contain other amino acids and phosphorus.However,there are few articles about L-lysine·H2SO4 product regarding intestinal morphology and liver pathology of broiler chickens.In this article,we focus on the absorption and metabolism of L-lysine·H2SO4 revealed in the variation of intestinal morphology and liver pathology to determine the tolerance of chicks for L-lysine·H2SO4.Methods: To evaluate the tolerance of broilers for L-lysine·H2SO4,240 one day old broilers were allocated randomly to one of five dietary treatments which included corn-soybean diets containing 0,1%,4%,7% or 10% L-lysine·H2SO4(L-lysine content = 55%).Results: Supplementation of 1% L-lysine·H2SO4 in the diet had no negative effects.However,4%,7% or 10% Llysine·H2SO4 supplementation produced negative responses on broiler performance,carcass characteristics,blood biochemistry,and particularly on intestinal morphology and liver pathology compared with broilers fed the control diet.Conclusion: Our results show that supplementation with 1% L-lysine·H2SO4 had no negative effects on performance,carcass characteristics,blood biochemistry,intestinal morphology and liver pathology in broilers,but supplementation with 4%,7% or 10% L-lysine·H2SO4 produced a negative response,particularly with respect to intestinal morphology and liver pathology.

Horizontal transfer of vanA between probiotic Enterococcus faecium and Enterococcus faecalis in fermented soybean meal and in digestive tract of growing pigs

Background: The aim of this study was to investigate the intergeneric transfer of vancomycin resistance gene vanA between probiotic enterococci in the fermentation progress of soybean meal and in the digestive tract of growing pigs.One vanA genotype vancomycin resistant E.faecium strain,Efm4,and one chloramphenicol-resistant E.faecalis strain,Efs2,were isolated from twenty-nine probiotic basis feed material/additive samples.For in vitro conjugation,Efm4 and Efs2 were used as starter to ferment soybean meal.For in vivo conjugation,thirty growing pigs were randomly assigned to five groups(n = 6),treated with a basic diet,or supplemented with 10% fermented soybean meal,1% Efm4,5% Efs2 or a combination of 1% Efm4 + 5% Efs2 for 7 d,respectively.Fecal samples of pigs in each group were collected daily for the isolation and dynamic analysis of Efm4,Efs2 and transconjugants.The sequence types(STs) of Efm4,Efs2 and transconjugants were analyzed by multilocus sequence typing(MLST).The vanA harboring plasmid in Efm4 and transconjugants was analyzed by S1-pulsed field gel electrophoresis(PFGE)and further verified by multiple alignments.Results: The results showed that,in FSBM,transconjugants were detected 1 h after the fermentation,with a conjugation frequency of ~ 10^-3 transconjugants/recipient.Transconjugants proliferated with Efm4 and Efs2 in the first 8 h and maintained steadily for 10 d till the end of the experiment.Additionally,in vivo experiment showed that transcojugants were recovered in one of six pigs in both FSBM and Efm4 + Efs2 groups,with conjugation frequency of ~ 10^-5 and ~ 10^-4,respectively.MLST revealed the ST of Efm4,Efs2 and transconjugants was ST1014,ST69 and ST69,respectively.S1-PFGE confirmed the existence of the vanA-harboring,142,988-bp plasmid,which was also a multi-drug resistant plasmid containing Tn1546-like transposon.Conclusions: The findings revealed the potential safety hazard existing in the commercial probiotic enterococci in China,because the horizontal transfer from farm to fork could potentially pose a safety risk to the public.

Leucine Supplementation in a Chronically Protein-Restricted Diet Enhances Muscle Weight and Postprandial Protein Synthesis of Skeletal Muscle by Promoting the mTOR Pathway in Adult Rats

Low protein intake causes a decrease in protein deposition in most animal tissues. The purpose of this study was to investigate whether leucine supplementation would increase the synthesis rate of protein and muscle weight in adult rats, which chronically consume only 58.8% of their protein requirements. Thirty-six male Sprague-Dawley rats were assigned to one of three dietary treatments including a 20% casein diet （CON）, a 10% casein ＋ 0.44% alanine diet （R）, and a 10% casein ＋ 0.87% leucine diet （RL）. After a 10 d dietary treatment, plasma amino acid levels were measured after feeding, the gastrocnemius muscles and soleus muscles were harvested and weighed, and the fractional synthesis rate （FSR） and mammalian target of rapamycin （mTOR） signaling proteins in skeletal muscle were measured. Regarding the plasma amino acid level, the RL group had the highest concentration of leucine （P 〈 0.05） and the lowest concentration of isoleucine （P 〈 0.05） among the three groups, and the CON group had a lower concentration of valine （P 〈 0.05） than the R and RL groups. Compared with the R and RE groups, the CON group diet significantly increased （P 〈 0.05） feed intake, protein synthesis rate, and the phosphorylation of eukaryutic initiation factor 4E binding protein 1 （4E-BP1）, and decreased the weight of abdominal adipose. Compared with the R group, the RL group significantly increased in gastrocnemius muscle weight, protein synthesis rate, and phosphorylation of both ribosomal protein $6 kinase 1 （56K1） and 4E-BP1. In conclusion, when protein is chronically restricted in adult rat diets, leucine supplementation moderately improves body weight gain and increases muscle protein synthesis through mTOR activation,

Different dietary starch patterns in lowprotein diets:effect on nitrogen efficiency,nutrient metabolism,and intestinal flora in growing pigs

Background:Protein releases amino acids faster than starch releases glucose in digestive tract of pigs fed lowprotein(LP)diets.Poor synchronization of dietary glucose and amino acids supply leads to compromised nitrogen efficiency.Dietary starch patterns modulation may improve this situation.Methods:Growing barrows(29.7±2.0 kg)were randomly allotted into 5 dietary treatments with LP diets consisting of different purified starches.Treatments included:waxy corn starch(W LP),corn starch+waxy corn starch(C+W LP),corn starch(C LP),pea starch+waxy corn starch(P+W LP)and pea starch(P LP).In the experiment,growth performance,protein deposition,nutrient metabolism,and fecal microbial community of pigs were investigated.In vitro starch digestion was used for predicting the in vivo glucose response.Results:Dietary starch in vitro glucose release profile was determined by starch source and the ratio of amylopectin and amylose.C+W LP treatment showed decreased total nitrogen excretion and plasma citrulline concentration and improved plasma leptin concentration among treatments(P<0.05).Besides,the highest nitrogen apparent biological value,whole-body protein deposition and growth performance and lowest urinary nitrogen excretion were also observed in C+W LP treatment.Compared with the other groups,C+W LP and C LP showed increased plasma pyruvate,IGF-1,and lipase concentrations(P<0.05).The W LP group presented dramatically increased plasma alanine and urea nitrogen concentration and decreased aldolase and leptin concentrations(P<0.05).Dietary starch patterns did not make an impact on bacterial richness and diversity,but changed the taxonomic and functional structures of the microbial communities.Microbial protein fermentation product(isobutyrate and isovalerate)presented increased in P LP treatments compared with the other treatments(P<0.05).Conclusions:Dietary starch patterns modulation can regulate dietary glucose release profile,nutrient metabolism,protein turnover,and fecal microbial fermentation in pigs.The optimal dietary glucose release profile effectively strengthened whole-body protein deposition and improve nitrogen efficiency and growth performance in growing pigs fed LP diets.

Progresses on Personalized Nutritional Evaluation and Recommendation

Health is maintained by a state of dynamic homeostasis in which nutrient intake and ex- penditure are of good balance. Therefore, it is important to know exactly the nutritional value of food sources, as well as the nutritional requirements of individuals, in order to achieve optimal nutrition. Considering the interaction between diet and individual back- ground, nutritional evaluation and recommendation has become a complicate issue needing further investigations. While traditional nutrition research has made significant progress in population nutrition, modern nutrition research is now becoming possible to focus on personalized nutrition in health promotion, disease prevention, performance improvement, and risk assessment of individual with the development of emerging omics technologies. This review tried to summarize the methods used in nutritional evaluation and recom- mendation as well as their applications. Though personal nutrition evaluation and recommendation are still not well-established, utilization of these advanced technologies may expand our knowledge in bioavailability and bioefficacy of diet ingredients, pathophysiological changes in response to dietary intervention, as well as nutrition-associated disease biomarkers discovery, and thus contributing to personalized nutrition.

Advances in research on solid-state fermented feed and its utilization:The pioneer of private customization for intestinal microorganisms

With sustainable development of biotechnology,increasing attention has been placed on utilization of solid-state fermented feed(SFF).Solid-state fermented feed has been a candidate strategy to alleviate the contradiction between supply and demand of feed resources,ensure food hygiene safety,promoting energy conservation,and emission reduction.In production of SFF,a variety of organic acids,enzymes,vitamins,peptides,and other unknown growth factors are produced,which could affect performance of animals.Solid-state fermented feed produced by different fermentation techniques has great instability on different physiological stages of different animals,which hinders the application and standardized production of SFF.Herein,we summarize the current advances in the role of the characteristics of SFF prepared by different manufacturing technique and its research progress in animal experiments on growth performance,gastrointestinal ecology,and immune system,so as to provide references for further acquiring a relatively perfect set of SFF production and evaluation systems.

Dietary modulation of endogenous host defense peptide synthesis as an alternative approach to in-feed antibiotics

Traditionally, antibiotics are included in animal feed at subtherapeutic levels for growth promotion and disease prevention.However, recent links between in-feed antibiotics and a rise in antibiotic-resistant pathogens have led to a ban of all antibiotics in livestock production by the European Union in January 2006 and a removal of medically important antibiotics in animal feeds in the United States in January 2017.An urgent need arises for antibiotic alternatives capable of maintaining animal health and productivity without triggering antimicrobial resistance.Host defense peptides(HDP) are a critical component of the animal innate immune system with direct antimicrobial and immunomodulatory activities.While in-feed supplementation of recombinant or synthetic HDP appears to be effective in maintaining animal performance and alleviating clinical symptoms in the context of disease, dietary modulation of the synthesis of endogenous host defense peptides has emerged as a cost-effective,antibiotic-alternative approach to disease control and prevention.Several different classes of smallmolecule compounds have been found capable of promoting HDP synthesis.Among the most efficacious compounds are butyrate and vitamin D.Moreover, butyrate and vitamin D synergize with each other in enhancing HDP synthesis.This review will focus on the regulation of HDP synthesis by butyrate and vitamin D in humans, chickens, pigs, and cattle and argue for potential application of HDP-inducing compounds in antibiotic-free livestock production.

Effectiveness and safety evaluation of graded levels of N-carbamylglutamate in growing-finishing pigs

The aim of this study was as follows:1)to investigate the effects of graded levels of N-carbamylglutamate(NCG)on performance,blood biochemical indexes,carcass traits and related indicators in growingfinishing pigs,and 2)to determine the optimal supplemental level.The toxicity of high-dose(much higher than recommended levels)NCG was assessed by routine blood tests and blood biochemical and histopathologic examinations of the heart,liver,spleen,lung,kidney and stomach.One hundred and forty-four growing-finishing pigs(DurocLarge WhiteLandrace,32.24±1.03 kg)were used in a 74-d experiment and each treatment was replicated 6 times with 4 pigs(2 barrows and 2 gilts)per replicate.The dietary treatments were a corn-soybean meal basal diet supplemented with 0%(control),0.05%,0.1%,0.15%,0.2%or 1%NCG.The first 5 groups were used to explore the optimal supplemental level of NCG,while the control,0.1%and 1%NCG groups were used to explore the safety of high-dose NCG.Compared with the normal control group,the final body weight and average daily gain tended to be higher in the 0.1%group(P=0.08),the lean percentage tended to be higher in the 0.05%group(P=0.07),the levels of free amino acids in the blood significantly increased in the 0.1%group(P<0.05),both 0.1%and 0.15%NCG supplementation increased the levels of nitric oxide(NO)in serum(P=0.07)and muscle growth-and lipid metabolism-related gene expression(P<0.05)and NCG supplementation improved C18:1N9C monounsaturated fatty acids(MUFA)in a dose-dependent manner(P=0.08).In addition,routine blood tests,blood biochemical indexes and histopathological examination revealed no abnormalities.Overall,increasing the levels of NCG did not linearly improve the above indicators;the 0.1%dose showed the best effect,and a high dose(1%)did not pose a toxicity risk.