Effects of Supplemental Glutamine and Lysine on Growth Performance of Broiler Chickens

The optimum levels of Lysine and Glutamine needed for growth performance and maintenance of the chicken broilers were evaluated in a randomized 3 × 4 factorial arrangement of dietary treatments. The battery cages measured 99 × 66 × 25 cm that can be sufficient for 5 birds. Day old Chicken broilers totaling 180 were assigned to dietary treatments comprising of 3 concentrations of Lysine (0.85, 1.14, and 1.42) each in combination with 4 concentrations of Glutamine (0, 1, 2, and 3). Each dietary treatment was replicated 3 times and each replication had 5 birds. The birds were given feed and water ad libitum with a 23-hour light regimen for a period of 4 weeks. Then, the experimental birds were evaluated for body weight gain, feed consumption, and feed conversion in order to determine their optimum requirement for dietary Lysine and Glutamine. Based on the findings of this study, the highest performance was observed in birds fed the diet supplemented with 1.42 lysine and 1% glutamine, but the highest improvement in feed conversion was observed in diet contain 1.14 and 1.42 with 1% and 3% glutamine, respectively. Birds fed 1.42 lysine and 1% glutamine had the highest total body weight gain and feed consumption. The lysine requirements in the diet for Chicken are between 1.14 and 1.42 with glutamine level of 1%.

Dietary Macleaya cordata extract supplementation improves the growth performance and gut health of broiler chickens with necrotic enteritis

Background The poultry industry needs effective antibiotic alternatives to control outbreaks of necrotic enteritis(NE)caused by Clostridium perfringens.Methods The aim of this study was to investigate the effects of dietary supplementation with Macleaya cordata extract(MCE)on the immune function and gut microbiota of broilers with NE.A total of 2881-day-old broiler chicks were randomly assigned to a 2×2 factorial arrangement with two concentrations of dietary MCE supplementation(0 or 350 mg/kg of diet)and two disease challenge statuses(control or NE).Results The results revealed that NE significantly increased the feed conversion rate(FCR),mortality,intestinal lesion score,the levels of IL-1β,IL-17 and IFN-γ/IL-4 in serum and IL-17/IL-10 in the jejunal mucosa,m RNA levels of TLR2,IFN-γand p Ig R in the jejunum,and Clostridium perfringens concentrations in the cecum.NE significantly decreased the body weight(BW),body weight gain(BWG),jejunal villus height,V/C,m RNA level of AMPK-α1 in jejunum,IL-4 level in the jejunal mucosa and lactic acid bacteria abundance in the cecum.MCE significantly increased BW,BWG,jejunal villus height,V/C,m RNA levels of occludin,ZO-1 and AMPK-α1 in the jejunum,the levels of Ig A and Ig G in serum and IL-10 in the jejunal mucosa and m RNA levels of NF-κB,IL-10 and MHC-II in the jejunum.Additionally,MCE significantly decreased the FCR,mortality,intestinal lesion score,jejunal crypt depth,the levels of IFN-γand IL-17 in serum and IL-17/IL-10 in the jejunal mucosa,Clostridium perfringens concentrations in the cecum,and m RNA levels of IL-17/IL-10 in the jejunum.Moreover,NE significantly increased the abundance of bacteria that are associated with inflammation,obesity and depression(Alistipes,Barnesiella,Intestinimonas,RF39 and UCG-005)and significantly decreased the abundance of short-chain fatty acid(SCFA)-producing bacteria(Anaerotruncus,Butyricicoccus and Bacteroides)in the cecum.MCE significantly increased the abundance of SCFA-producing bacteria(Streptococcus,Ruminococcus_torques_group and Lachnospiraceae_NK4A136_group)and significantly reduced the abundance of bacteria that are associated with inflammation and obesity(Alistipes,Barnesiella and UCG-010)in the cecum.In the cecum of broilers with NE,the relative abundance of Barnesiella and Alistipes was higher and that of Lachnoclostridium and Shuttleworthia was lower.Interestingly,these trends were reversed by the addition of MCE to the diet.Spearman correlation analysis showed that Barnesiella and Alistipes were associated with enhanced intestinal inflammation and inhibited growth performance,whereas Lachnoclostridium and Shuttleworthia were associated with anti-inflammatory effects.Conclusions MCE ameliorated the loss of growth performance in broiler chickens with NE,probably by regulating the intestinal barrier,immune function,and gut microbiota.

Pretreatment with probiotics Enterococcus faecium NCIMB 11181 attenuated Salmonella Typhimurium-induced gut injury through modulating intestinal microbiome and immune responses with barrier function in broiler chickens

Background:Preventing Salmonella infection and colonization in young birds is key to improving poultry gut health and reducing Salmonella contamination of poultry products and decreasing salmonellosis for human consumption(poultry meat and eggs).Probiotics can improve poultry health.The present study was conducted to investigate the impact of a probiotics,Enterococcus faecium NCIMB 11181(E.faecium NCIMB 11181)on the intestinal mucosal immune responses,microbiome and barrier function in the presence or absence of Salmonella Typhimurium(S.Typh-imurium,ST)infection.Methods:Two hundred and forty 1-day-old Salmonella-free male broiler chickens(Arbor Acres AA+)were randomly allocated to four groups with 6 replicate cages of 10 birds each.The four experimental groups were follows:(1)nega-tive control(NC),(2)S.Typhimurium,challenged positive control(PC),(3)the E.faecium NCIMB 11181-treated group(EF),(4)the E.faecium NCIMB 11181-treated and S.Typhimurium-challenged group(PEF).Results:Results indicated that,although continuous feeding E.faecium NCIMB 11181 did not obviously alleviate growth depression caused by S.Typhimurium challenge(P>0.05),E.faecium NCIMB 11181 addition significantly blocked Salmonella intestinal colonization and translocation(P<0.05).Moreover,supplemental E.faecium NCIMB 11181 to the infected chickens remarkably attenuated gut morphological structure damage and intestinal cell apoptosis induced by S.Typhimurium infection,as evidenced by increasing gut villous height and reducing intes-tinal TUNEL-positive cell numbers(P<0.05).Also,E.faecium NCIMB 11181 administration notably promoting the production of anti-Salmonella antibodies in intestinal mucosa and serum of the infected birds(P<0.05).Addition-ally,16S rRNA sequencing analysis revealed that E.faecium NCIMB 11181 supplementation ameliorated S.Typhimu-rium infection-induced gut microbial dysbiosis by enriching Lachnospiracease and Alistipes levels,and suppressing Barnesiella abundance.Predicted function analysis indicated that the functional genes of cecal microbiome involved in C5-branched dibasic acid metabolism;valine,leucine and isoleucine biosynthesis;glycerolipid metabolism and lysine biosynthesis were enriched in the infected chickens given E.faecium NCIMB 11181.While alanine,asparate and glutamate metabolism;MAPK signal pathway-yeast;ubiquine and other terpenoid-quinore biosynthesis,protein processing in endoplasmic reticulum;as well as glutathione metabolism were suppressed by E.faecium NCIMB 11181 addition.Conclusion:Collectively,our data suggested that dietary E.faecium NCIBM 11181 supplementation could ameliorate S.Typhimurium infection-induced gut injury in broiler chickens.Our findings also suggest that E.faecium NCIMB 11181 may serve as an effective non-antibiotic feed additive for improving gut health and controlling Salmonella infection in broiler chickens.

Growth performance,nutrient digestibility,intestinal morphology,cecal mucosal cytokines and serum antioxidant responses of broiler chickens to dietary enzymatically treated yeast and coccidia challenge

Background There is a growing search for natural feed additives to alleviate the deleterious effects of coccidia infection in poultry production.This study aimed to investigate the effect of enzymatically treated yeast(ETY) on the growth performance,nutrient digestibility,intestinal morphology,antioxidative status,and cecal mucosa cytokines of coccidia-challenged broiler chickens.Methods From d 1 to 14 post hatching,480 broiler chickens were allocated to 3 corn-soybean meal-based experimental diets with increasing concentrations of ETY(0,1,or 2 g/kg).The experiment was designed as a randomized complete block design with body weight(BW) used as a blocking factor.On d 14 post hatching,the birds were re-randomized within each of the 3 experimental diets.Each of the 3 diet groups was split into a challenge or no-challenge group.This resulted in a 3 × 2 factorial arrangement of treatments.The coccidia challenge was administered on d 15 by an oral gavage.Results Dietary ETY improved(P < 0.05) the G:F of birds on d 21 regardless of the challenge state and linearly increased(P < 0.01) the apparent ileal digestibility of dry matter(DM),nitrogen,and gross energy(GE).The coccidia challenge decreased(P < 0.05) BW gain and feed intake of broiler chickens and reduced(P < 0.01) the total tract retention of DM,GE,and nitrogen.The coccidia challenge increased(P < 0.01) the mRNA gene expression of TNFα,IL-1β,IL-10,and IL-6 in the cecal mucosa.There was a tendency(P ne = 0.08) for ETY to linearly reduce IL-1β expression.Additionally,ETY supplementation increased(P < 0.05) the geexpression of OCLN.Serum catalase increased(P < 0.05) with dietary ETY in broiler chickens on d 21.Dietary ETY linearly increased(P < 0.05) the ileal villus height to crypt depth ratio,and ileal goblet cell density in broiler chickens.The ileal and excreta oocyst counts decreased(P < 0.01) with increasing supplementation of dietary ETY in coccidia-challenged broiler chickens on d 21.Conclusions Dietary ETY enhanced nutrient utilization and augmented intestinal development in broiler chickens.However,dietary ETY did not completely attenuate the adverse effects of a coccidia challenge in broiler chickens.

Effects of rearing system and antibiotic treatment on immune function,gut microbiota and metabolites of broiler chickens

Background:In China,cage systems with a high space utilization have gradually replaced ground litter systems,but the disease incidence of chickens in cages is higher.Broilers in the ground litter pens may be stimulated by more environmental microbes during the growth process and show strong immune function and status,but knowledge of which microbes and their metabolites play an immunomodulatory role is still limited.This study aimed to explore the differences and correlations in the immune function,gut microbiota and metabolites and the importance of gut microbiota of broilers raised in cages and ground litter pens.Methods:The experiment involved a 2×2 factorial arrangement,with rearing systems(cages or ground litter pens)and antibiotic treatment(with or without broad-spectrum antibiotics in drinking water)as factors.Results:The results showed that,compared with the cage group,the ground litter broilers had stronger nonspecific immune function(Macrophages%and NO in blood),humoral immune function(IgG in blood,LPS stimulation index in ileum)and cellular immune function(T%,Tc%,ConA stimulation index and cytokines in blood).Antibiotic(ABX)treat-ment significantly reduced nonspecific immune function(Macrophages%and NO in blood,iNOS and Mucin2 mRNA expression in ileum),humoral immune function(IgG in blood and sIgA in ileum)and cellular immune function(T%and cytokines in blood,Th and Tc ratio,TLRs and cytokines mRNA expression in ileum).Furthermore,the ground litter broil-ers had higherαdiversity of microbiota in ileum.The relative abundance of Staphylococcus,Jeotgalicoccus,Jeotgalibaca and Pediococcus in the ileum of ground litter broilers were higher.ABX treatment significantly reduced theαdiversity of ileal microbiota,with less Chloroplast and Mitochondria.In addition,the levels of acetic acid,isobutyric acid,kynurenic acid and allolithocholic acid in the ileum of ground litter broilers were higher.Spearman correlation analysis showed that Jeotgalibaca,Pediococcus,acetic acid,kynurenic acid and allolithocholic acid were related to the immune function.Conclusions:There were more potential pathogens,litter breeding bacteria,short-chain fatty acids,kynurenine,allolithocholic acid and tryptophan metabolites in the ileum of broilers in ground litter pens,which may be the reason for its stronger immune function and status.

Dietary supplemental coated essential oils and organic acids mixture improves growth performance and gut health along with reduces Salmonella load of broiler chickens infected with Salmonella Enteritidis

Background Reducing Salmonella infection in broiler chickens by using effective and safe alternatives to antibiotics is vital to provide safer poultry meat and minimize the emergence of drug-resistant Salmonella and the spread of salmonellosis to humans.This study was to first evaluate the protective efficacy of feeding coated essential oils and organic acids mixture(EOA)on broiler chickens infected with Salmonella Enteritidis(S.Enteritidis,SE),and then its action mechanism was further explored.Methods A total of 4801-day-old Arbor Acres male chickens were randomly assigned into five treatments with six replicates,including non-challenged control fed with basal diet(A),SE-challenged control(B),and SE-infected birds fed a basal diet with 300 mg/kg of EOA(BL),500 mg/kg of EOA(BM)and 800 mg/kg of EOA(BH),respectively.All birds on challenged groups were infected with Salmonella Enteritidis on d 13.Results Feeding EOA showed a reversed ability on negative effects caused by SE infection,as evidenced by decreasing the feed conversion rate(FCR)and the ratio of villus height to crypt depth(VH/CD)(P<0.05),obviously decreasing intestinal and internal organs Salmonella load along with increasing cecal butyric acid-producing bacteria abundance(P<0.05).Moreover,supplemental different levels of EOA notably up-regulated claudin-1(CLDN-1),occludin(OCLN),zonula occludens-1(ZO-1),mucin-2(MUC-2),fatty acid binding protein-2(FABP-2),nuclear factor kappa-light-chainenhancer of activated B cells(NF-κB),myeloid differential protein-88(MyD88)and interleukin-6(IL-6)mRNA levels in the ileum of the infected chickens after challenge,whereas down-regulated toll-like receptor-4(TLR-4)mRNA levels(P<0.05).Linear discriminant analysis combined effect size measurements analysis(LEfSe)showed that the relative abundance of g_Butyricicoccus,g_Anaerotruncus and g_unclassified_f_Bacillaceae significantly was enriched in infected birds given EOA.Also,phylogenetic investigation of communities by reconstruction of unobserved states(PICRUSt)analysis showed that alpha-linolenic acid metabolism,fatty acid metabolism and biosynthesis of unsaturated fatty acids were significantly enriched in the EOA group.Conclusion Our data suggest that the essential oils and organic acids mixture can be used as an effective strategy to ameliorate and alleviate Salmonella Enteritidis infection in broilers.

Intoxication Induced by Urea Containing Diets in Broiler Chickens: Effect on Weight Gain, Feed Conversion Ratio, Hematological and Biochemical Profiles

Urea as a source of cheap non-protein nitrogen is used to adulterate fish and meat meals which are basic components of broiler diets. The present study was carried out to elucidate the effects of urea on weight gain, and hematological and biochemical profiles. A total of 48 broiler chicks were randomly allotted into 4 groups, designated Groups 1, 2, 3 and 4 of 12 birds each. Birds in Groups 2, 3 and 4 were fed on diets containing urea at the levels of 1%, 2.5% and 4%, respectively. Birds in Group 1 served as control and were not exposed to urea. Experimentation period was for 3 weeks and experiment was terminated when birds were 42 days of age. Body weight of all intoxicated birds at the various intervals was significantly decreased in comparison with that of the untreated control. Compared with control, all intoxicated broilers manifested significant (P ≤ 0.05) decrease in all hematological parameters involving erythrocytic and total leucocytic counts, Hemoglobin (Hb) and Packed Cell Volume (PCV) on a dose- and time-pattern. In comparison with the control levels, biochemical profile of the intoxicated birds disclosed significant decrease in blood glucose level and significant increase in serum uric acid, urea, Alkaline Phosphatase (ALP) and Lactate Dehydrogenase (LDH) levels. Based upon the present data, it was concluded that the addition of urea to broiler diets bears serious sequences concerning the general health condition, performance, weight gain, and hematological and biochemical profiles.

Effects of tannic acid on the immunity and intestinal health of broiler chickens with necrotic enteritis infection

Background In broiler chickens,necrotic enteritis(NE)infection can reduce production performance.Tannic acid has shown great potential as a treatment of NE in broilers.However,the appropriate dosage of tannic acid in NE of broilers and the improvement effect on intestinal health are not very clear.In this study,we aimed to investigate the effects of different doses of tannic acid on the production performance,immunity,and intestinal health of broilers by constructing an NE model with C.perfringens infection and determining the appropriate dosage of tannic acid with regard to NE.Results Challenged birds showed significant reduction in body weight,villus height,and the ratio of villus height to crypt depth(P<0.05)and increase in the feed consumption gain ratio,intestinal lesion score,and crypt depth(P<0.05).The infection significantly reduced the relative Bacteroidota and Ligilactobacillus abundance(P<0.05)and increased the ratio of Firmicutes/Bacteroidota and cecal content of C.perfringens(P<0.05).Challenged birds fed diets supplemented with tannic acid showed significantly increased mRNA expression of nutrient transport carriers and intestinal barrier genes and growth performance and reduced serum zonulin and endotoxin levels(P<0.05).Addi-tion of tannic acid to the diet inhibited the inflammatory response by reducing the number of coccidia oocysts in feces and the content of C.perfringens in the cecum.Specifically,tannic acid reduced the serum levels of C reactive protein,myeloperoxidase,and specific IgY and ileal mucosal secretory immunoglobulin A levels in the ileal mucosa compared with those in the NE-infected birds.NE-infected birds fed diets supplemented with tannic acid also showed significantly increased relative Anaerocolumna,Thermoanaerobacterium,and Thermosinus abundance(P<0.05);their microbial composition and functional predictions were similar to those of the NC group.Conclusions Tannic acid in the diet alleviated NE by enhancing the intestinal barrier and absorption function.The recommended dietary tannic acid additive level is 500–750 mg/kg.Our study findings would be useful in reducing related economic losses in the broiler industry.

Encapsulated peracetic acid as a valid broad‑spectrum antimicrobial alternative,leading to beneficial microbiota compositional changes and enhanced performance in broiler chickens

Background Antimicrobial alternatives are urgently needed,including for poultry production systems.In this study,we tested the potential broad-range antimicrobial alternative peracetic acid,delivered in feed via the hydrolysis of encapsulated precursors through a 28-day study using 375 Ross 308 broiler chickens.We tested two peracetic acid concentrations,30 and 80 mg/kg on birds housed on re-used litter,and we evaluated the impact of both levels on gut microbial communities,bacterial concentration,antimicrobial resistance genes relative abundance and growth performance when compared to control birds housed on either clean or re-used litter.Results Body weight gain and feed conversion ratio improved in peracetic acid fed birds.At d 28,birds given 30 mg/kg of peracetic acid had a decreased Firmicutes and an increased Proteobacteria abundance in the jejunum,accompanied by an increase in Bacillus,Flavonifractor and Rombustia in the caeca,and a decreased abundance of tetracycline resistance genes.Chicken given 80 mg/kg of peracetic acid had greater caecal abundance of macrolides lincosamides and streptogramins resistance genes.Growth performance on clean litter was reduced compared to reused litter,which concurred with increased caecal abundance of Blautia,decreased caecal abundance of Escherichia/Shigella,Anaerostipes and Jeotgalicoccus,and greater gene abundance of vancomycin,tetracycline,and macrolides resistance genes.Conclusions Peracetic acid could be used as a safe broad-spectrum antimicrobial alternative in broilers.Encapsulated precursors were able to reduce the bacterial concentration in the jejunum whilst promoting the proliferation of probiotic genera in the caeca,especially at the low peracetic acid concentrations tested,and improve growth performance.Moreover,our findings offer further insights on potential benefits of rearing birds on re-used litter,suggesting that the latter could be associated with better performance and reduced antimicrobial resistance risk compared to clean litter rearing.

Feeding broiler chickens with arginine above recommended levels:effects on growth performance,metabolism,and intestinal microbiota

Background Arginine is an essential amino acid for chickens and feeding diets with arginine beyond the recommended levels has been shown to influence the growth performance of broiler chickens in a positive way.Nonetheless,further research is required to understand how arginine supplementation above the widely adopted dosages affects metabolism and intestinal health of broilers.Therefore,this study was designed to assess the effects of arginine supplementation(i.e.,total arginine to total lysine ratio of 1.20 instead of 1.06-1.08 recommended by the breeding company) on growth performance of broiler chickens and to explore its impacts on the hepatic and blood metabolic profiles,as well as on the intestinal microbiota.For this purpose,630 one-day-old male Ross 308 broiler chicks were assigned to 2 treatments(7 replicates each) fed a control diet or a crystalline L-arginine-supplemented diet for 49 d.Results Compared to control birds,those supplemented with arginine performed significantly better exhibiting greater final body weight at D49(3778 vs.3937 g;P < 0.001),higher growth rate(76.15 vs.79.46 g of body weight gained daily;P < 0.001),and lower cumulative feed conversion ratio(1.808 vs.1.732;P < 0.05).Plasma concentrations of arginine,betaine,histidine,and creatine were greater in supplemented birds than in their control counterparts,as were those of creatine,leucine and other essential amino acids at the hepatic level.In contrast,leucine concentration was lower in the caecal content of supplemented birds.Reduced alpha diversity and relative abundance of Firmicutes and Proteobacteria(specifically Escherichia coli),as well as increased abundance of Bacteroidetes and Lactobacillus salivarius were found in the caecal content of supplemented birds.Conclusions The improvement in growth performance corroborates the advantages of supplementing arginine in broiler nutrition.It can be hypothesized that the performance enhancement found in this study is associated with the increased availability of arginine,betaine,histidine,and creatine in plasma and the liver,as well as to the ability of extra dietary arginine to potentially ameliorate intestinal conditions and microbiota of supplemented birds.However,the latter promising property,along with other research questions raised by this study,deserve further investigations.

Effects of dietary supplementation of probiotic,Clostridium butyricum,on growth performance,immune response,intestinal barrier function,and digestive enzyme activity in broiler chickens challenged with Escherichia coli K88

Background： Colibacillosis caused by enterotoxigenic Escherichia coil （E. coil} results in economic losses in the poultry industry. Antibiotics are usually used to control colibacillosis, however, E. coli has varying degrees of resistance to different antibiotics. Therefore the use of probiotics is becoming accepted as an alternative to antibiotics. In this study, we evaluated the effects of Clostfidium butyricum （C. butyficum） on growth performance, immune response, intestinal barrier function, and digestive enzyme activity in broiler chickens challenged with Eschefichia coli （E. coil） K88. Methods： The chickens were randomly divided into four treatment groups for 28 days. Negative control treatment （NC） consisted of birds fed a basal diet without E. coil K88 challenge and positive control treatment （PC） consisted of birds fed a basal diet and challenged with E. coil K88. C. buO/ricum probiotic treatment （CB） consisted of birds fed a diet containing 2 x 107 cfu C. buO/ricum/kg of diet and challenged with E. coil K88. Colistin sulfate antibiotic treatment （CS） consisted of birds fed a diet containing 20 mg colistin sulfate/kg of diet and challenged with E. coil K88. Results： The body weight （BW） and average day gain （ADG） in the broilers of CB group were higher （P 〈 0.05） than the broilers in the PC group overall except the ADG in the 14-21 d post-challenge. The birds in CB treatment had higher （P 〈 0.05） concentration of tumor necrosis factor-a （TNF-a） at 3 and 7 d post-challenge, and higher （P 〈 0.05） concentration of interleukin-4 （IL-4） at 14 d post-challenge than those in the PC treatment group. The concentration of serum endotoxin in CB birds was lower （P 〈 0.05） at 21 d post-challenge, and the concentrations of serum diamine oxidase in CB birds were lower （P 〈 0.05） at 14 and 21 d post-challenge than in PC birds. Birds in CB treatment group had higher （P 〈 0.05） jejunum villi height than those in PC, NC, or CS treatment at 7, 14, and 21 d post-challenge. In comparison to PC birds, the CB birds had lower （P 〈 0.05） jejunum crypt depth during the whole experiment. The birds in CB or CS treatment group had higher （P 〈 0.05） activities of amylase and protease at 3, 7, and 14 d post-challenge, and higher （P 〈 0.05） activity of lipase at 3, 7 d post-challenge than PC birds.

In vitro antibacterial activity of thymol and carvacrol and their effects on broiler chickens challenged with Clostridium perfringens

Background： In the post-antibiotic era, essential oils （EO） are promising alternatives to growth-promoting antibiotics. The aim of the present study was to investigate the antibacterial activities of an EO product and its components thymol and carvacrol in vitro, and the efficacy of EO to control Clostridium perffingens challenge in broiler chickens. Results： The in vitro minimum inhibitory concentration assay showed strong antibacterial activity of the EO product, thymol, and carvacrol against pathogenic Escherichia col~, C. perffingens, and Salmonella strains, and weak activity towards beneficial Lactobacillus strains. Besides, an additive effect was observed between thymol and carvacrol. The in vivo study was carried out with 448 male broiler chicks following a 4 x 2 factorial arrangement to test the effects of EO supplementation （0, 60, 120, or 240 mg/kg EO in wheat-based diet）, pathogen challenge （with or without oral gavage of C. perfr/ngens from day 14 to day 20） and their interactions. Each treatment consisted of eight replicate pens （seven birds/pen）. The challenge led to macroscopic gut lesions, and resulted in a significant increase in ileal populations of C. perffingens and Eschefichia subgroup （P ≤ 0.05） on day 21. Dietary EO supplementation did not influence C. perfringens numbers, but linearly alleviated intestinal lesions on day 21 and 28 （P= 0.010 and 0.036, respectively）, and decreased Eschefichia populations in ileum with increased EO dosages （P = 0.027 and 0.071 for day 21 and 28, respectively）. For caecum, EO quadratically influenced Lactobacillus populations on day 21 （P = 0.002）, and linearly decreased the numbers of total bacteria and Eschefichio on day 28 （P = 0.026 and 0.060, respectively）. Mean thymol and carvacrol concentrations in the small intestine were 0.21 and 0.20 μg/g in intestinal digesta （wet weight）, respectively, for birds fed 60 mg/kg EO, and 0.80 and 0.71 μg/g, respectively, for birds fed 240 mg/kg EO. Conclusions： These results indicated that dietary EO supplementation could affect intestinal microbiota and alleviate intestinal lesions in broilers, which may contribute in controlling C. perffingens infection in broiler chickens.

Progress towards reduced-crude protein diets for broiler chickens and sustainable chicken-meat production

The prime purpose of this review is to explore the pathways whereby progress towards reduced-crude protein(CP)diets and sustainable chicken-meat production may be best achieved.Reduced-CP broiler diets have the potential to attenuate environmental pollution from nitrogen and ammonia emissions;moreover,they have the capacity to diminish the global chicken-meat industry’s dependence on soybean meal to tangible extents.The variable impacts of reduced-CP broiler diets on apparent amino acid digestibility coefficients are addressed.The more accurate identification of amino acid requirements for broiler chickens offered reduced-CP diets is essential as this would diminish amino acid imbalances and the deamination of surplus amino acids.Deamination of amino acids increases the synthesis and excretion of uric acid for which there is a requirement for glycine,this emphasises the value of so-called“non-essential”amino acids.Starch digestive dynamics and their possible impact of glucose on pancreatic secretions of insulin are discussed,although the functions of insulin in avian species require clarification.Maize is probably a superior feed grain to wheat as the basis of reduced-CP diets;if so,the identification of the underlying reasons for this difference should be instructive.Moderating increases in starch concentrations and condensing dietary starch:protein ratios in reduced-CP diets may prove to be advantageous as expanding ratios appear to be aligned to inferior broiler performance.Threonine is specifically examined because elevated free threonine plasma concentrations in birds offered reduced-CP diets may be indicative of compromised performance.If progress in these directions can be realised,then the prospects of reduced-CP diets contributing to sustainable chicken-meat production are promising.

Dietary encapsulated essential oils and organic acids mixture improves gut health in broiler chickens challenged with necrotic enteritis

Background:The poultry industry is in need of effective antibiotic alternatives to control outbreaks of necrotic enteritis(NE)due to Clostridium perfringens.In the present study,we investigated the effects of dietary supplementation with a blend of encapsulated essential oils and organic acids(BLJ)on growth performance and gut health using a coinfection model of NE in broiler chickens.Methods:Two hundred and eighty-eight one-day-old male Arbor Acres broiler chicks were randomly assigned using a 2×2 factorial design into two groups fed either 0 or 500 mg/kg dietary BLJ and co-challenged(or not challenged for the control)with Eimeria spp./C.perfringens.Results:Infected birds fed the BLJ-supplemented diet exhibited an improved feed conversion ratio throughout the trial(P<0.01),a higher villus height and villus height/crypt depth ratio,and reduced intestinal C.perfringens counts,liver C.perfringens carriage,gut lesion scores and serum fluorescein isothiocyanate dextran(FITC-D)concentrations at 7 d postinfection compared with those of birds without BLJ supplementation(P<0.05).NE-infected birds fed BLJ exhibited significantly upregulated claudin-1 and IGF-2 mRNA levels(P<0.05),increased A20 mRNA expression and significantly downregulated TRAF-6,TNFSF15 and TOLLIP mRNA levels in the jejunum at 7 d post-infection compared with those in birds without BLJ supplementation(P<0.05).Compared with the uninfected and untreated birds,the uninfected birds fed BLJ displayed increased relative abundances of Lactobacillus and Coprococcus but reduced Rikenellaceae levels.Compared with the unsupplemented NE-challenged birds,infected birds fed BLJ showed an increased relative abundance of Unclassified_Lachnospiraceae and a significantly decreased relative abundance of Erysipelotrichaceae.Conclusion:BLJ supplementation improved growth performance and gut health in NE-infected broiler chickens by strengthening the intestinal barrier function,positively modulating the gut microbiota community and differentially regulating intestinal immune responses.Our results also suggested that adding BLJ effectively controlled NE infections after experimental Eimeria and Clostridium perfringens coinfection.

Molecular mechanisms of growth depression in broiler chickens(Gallus Gallus domesticus)mediated by immune stress:a hepatic proteome study

Background:Immunological stress decreases feed intake,suppresses growth and induces economic losses.However,the underlying molecular mechanism remains unclear.Label-free liquid chromatography and mass spectrometry(LC-MS)proteomics techniques were employed to investigate effects of immune stress on the hepatic proteome changes of Arbor Acres broilers(Gallus Gallus domesticus)challenged with Escherichia coli lipopolysaccharide(LPS).Results:Proteomic analysis indicated that 111 proteins were differentially expressed in the liver of broiler chickens from the immune stress group.Of these,28 proteins were down-regulated,and 83 proteins were up-regulated in the immune stress group.Enrichment analysis showed that immune stress upregulated the expression of hepatic proteins involved in defense function,amino acid catabolism,ion transport,wound healing,and hormone secretion.Furthermore,immune stress increased valine,leucine and isoleucine degradation pathways.Conclusion:The data suggests that growth depression of broiler chickens induced by immune stress is triggered by hepatic proteome alterations,and provides a new insight into the mechanism by which immune challenge impairs poultry production.

Bacteriophage cocktail supplementation improves growth performance,gut microbiome and production traits in broiler chickens

Background:Effective antibiotic alternatives are urgently needed in the poultry industry to control disease outbreaks.Phage therapy mainly utilizes lytic phages to kill their respective bacterial hosts and can be an attractive solution to combating the emergence of antibiotic resistance in livestock.Methods:Five hundred and four,one-day-old broilers(Ross 308)were allotted to 1 of 4 treatment groups in a completely randomized design.Treatments consisted of CON(basal diet),PC(CON+0.025%Avilamax®),BP 0.05(CON+0.05%bacteriophage),and BP 0.10(CON+0.10%bacteriophage).Results:A significant linear effect on body weight gain(BWG)was observed during days 1–7,days 22–35,and cumulatively in bacteriophage(BP)supplemented groups.The BWG tended to be higher(P=0.08)and the feed intake(FI)was increased(P=0.017)in the PC group over CON group.A greater(P=0.016)BWG and trends in increased FI(P=0.06)were observed in the experiment in birds fed PC than CON diet.At the genus level,the relative abundance of Lactobacillus was decreased in PC(65.28%),while it was similar in BP 0.05 and BP 0.10(90.65%,86.72%)compared to CON(90.19%).At the species level,the relative abundance of Lactobacillus salivarus was higher in BP 0.05(40.15%)and BP 0.10(38.58%)compared to the CON(20.04%)and PC(18.05%).A linear reduction in the weight of bursa of Fabricius(P=0.022)and spleen(P=0.052)was observed in birds fed graded level of BP and an increase(P=0.059)in the weight of gizzard was observed in birds fed PC over BP diets.Linear and quadratic responses were observed in redness of breast muscle color in birds fed graded level of BP.Conclusions:The inclusion of the 0.05%and 0.1%BP cocktail linearly improved broiler weight during the first 7 days,22–35 days and cumulatively,whereas 0.05%BP addition was sufficient for supporting immune organs,bursa and spleen as well as enhancing gut microbiome,indicating the efficacy of 0.05%BP as a substitute antibiotic growth promoter in broiler diets.

Influence of REV and ALV-J Co-Infection on Immunologic Function of T Lymphocytes and Histopathology in Broiler Chickens

The aim of present investigation is to study the effect of single- and co-infection with REV and ALV-J on T lymphocytes bioactivities and histopathology in broiler chickens. The bioactivities of blood and spleen T lymphocytes including lymphoproliferation responses, cytotoxicitic responses, and histopathology of spleen were detected in broiler chickens singly- or co-infected with REV and ALV-J at different days post inoculation and the virus expressions in spleen of infected broiler chickens were detected with immunofluorescence assay （IFA）. The results indicated that blood and spleen T lymphocytes proliferation responses and cytotoxicity in broilers infected with REV or/and ALV-J were inhibited in the whole observed period compared with controls. In the co-infected chickens they were highly inhibited than in the single-infected. The histopathology of spleen in infected chickens at 17 and 37 d post inoculation （dpi） indicated that cell interium increased, the numbers of lymphocytes decreased, and the regrowth were destroyed or decreased, especially more significantly at 17 than at 37 dpi. The different numbers of virus were detected in spleen lymphocytes in REV- infected and/or ALV-J-infected chickens. In the spleen of co-infected chicken, both REV and ALV-J were detected and the total numbers of viruses were more than in chickens singly-infected with REV or ALV-J. Thus, the co-effect of REV and ALV-J caused more immunosuppression on T lymphocytes bioactivities in broiler chickens than single-effect of ALV-J or REV, which contributed to the sever histopathology and the product of tumor cells. This study will be helpful for understanding the effect of co-infection with many viruses and control them in poultry.

Effects of age on immune function in broiler chickens

Background:There are many diseases in poultry,many of which are caused by poor immune function.It is not clear how cytokines and various immune cell functions change with age in modern broilers.The purpose of this study was to explore the patterns of development of the immunity of the broiler chickens in cage.Results:The results showed that there were 3 development patterns of immunity in the broiler chickens.The first pattern was Down-Up.Cytokines and some immune indicators first decreased and then increased,and the lowest levels of immunity basically occurred from d 6 to 13.The second pattern was Up-Down,and from d 30 to 34,the highest levels of non-specific cellular immunity components,such as the peripheral blood mononuclear macrophage ratio,specific cellular immunity components,such as the peripheral blood helper T(Th)cell ratio and T cell and B cell proliferation activity,and mucosal immunity components,such as the ileal CD4,TGF-β1 and IgA mRNA levels,were observed.The third pattern was Up-Up,and the levels of the non-specific cellular immunity components,such as the serum nitric oxide(NO),C3 and C4 levels,the specific cellular immunity components,such as the spleen index,peripheral blood IL-2,IFN-γ/IL-4,cytotoxic T(Tc)cell ratio,and splenic NF-κB mRNA levels,the humoral immunity components,such as the serum IgG level,the mucosal immunity components,such as the ileal MHC-II,CD3d,TCRβsubunit,TCRζsubunit,IFN-γ,pIgR mRNA and ileal mucosa sIgA levels,were continuing to increase from d 1 to 34.Conclusions:It could be concluded that the immune system and its function have not developed well in the broiler chickens d 6 to 13 and that the immune system does not mature until d 30 to 34 in the broiler chickens in cages.It is necessary to enhance the immune function of the broiler chickens through nutritional measures from d 1 to 30.

Evaluation of curcumin and copper acetate against Salmonella Typhimurium infection,intestinal permeability,and cecal microbiota composition in broiler chickens

Background:Interest in the use of natural feed additives as an alternative to antimicrobials in the poultry industry has increased in recent years because of the risk of bacterial resistance.One of the most studied groups are polyphenolic compounds,given their advantages over other types of additives and their easy potentiation of effects when complexes are formed with metal ions.Therefore,the objective of the present study was to evaluate the impact of dietary supplementation of copper acetate(CA),curcumin(CR),and their combination(CA-CR)against Salmonella Typhimurium colonization,intestinal permeability,and cecal microbiota composition in broiler chickens through a laboratory Salmonella infection model.S.Typhimurium recovery was determined on day 10 post-challenge by isolating Salmonella in homogenates of the right cecal tonsil(12 chickens per group)on Xylose Lysine Tergitol-4(XLT-4)with novobiocin and nalidixic acid.Intestinal integrity was indirectly determined by the fluorometric measurement of fluorescein isothiocyanate dextran(FITC-d)in serum samples from blood obtained on d 10 post-S.Typhimurium challenge.Finally,microbiota analysis was performed using the content of the left caecal tonsil of 5 chickens per group by sequencing V4 region of 16S rRNA gene.Results:The results showed that in two independent studies,all experimental treatments were able to significantly reduce the S.Typhimurium colonization in cecal tonsils(CT,P<0.0001)compared to the positive control(PC)group.However,only CA-CR was the most effective treatment in reducing S.Typhimurium counts in both independent studies.Furthermore,the serum fluorescein isothiocyanate dextran(FITC-d)concentration in chickens treated with CR was significantly lower when compared to PC(P=0.0084),which is related to a decrease in intestinal permeability and therefore intestinal integrity.The effect of dietary treatments in reducing Salmonella was further supported by the analysis of 16S rRNA gene sequences using Linear discriminant analysis effect size(LEfSe)since Salmonella was significantly enriched in PC group(LDA score>2.0 and P<0.05)compared to other groups.In addition,Coprobacillus,Eubacterium,and Clostridium were significantly higher in the PC group compared to other treatment groups.On the contrary,Fecalibacterium and Enterococcus in CR,unknown genus of Erysipelotrichaceae at CA-CR,and unknown genus of Lachnospiraceae at CA were significantly more abundant respectively.Conclusions:CR treatment was the most effective treatment to reduce S.Typhimurium intestinal colonization and maintain better intestinal homeostasis which might be achieved through modulation of cecal microbiota.

Role of oregano and Citrus species-based essential oil preparation for the control of coccidiosis in broiler chickens

Background:Due to presence of drug-resistant Eimeria strains and raised public health safety concerns about drug residues in the meat,there is renewed interest in the search for natural alternatives to the coccidiosis control agents.This study was conducted to test the anticoccidial efficacy of oregano and Citrus spp.-based essential oils for broilers.Methods:A total of 2807-day-old broiler chicks were fed a control diet or diets with salinomycin or essential oils for up to 35 d of age.On d 14,half of the control groups and the treated groups were orally challenged with a coccidiosis vaccine at 25 times higher than the recommended vaccine dose.Control diet-fed chickens that were gavaged with phosphate-buffered saline were considered non-challenged control group.Results:Eimeria challenge or dietary additives failed to affect growth performance during the 7 to 20 d growth period although essential oil-fed chickens exhibited the lowest body wight gain(P=0.332)and the highest feed conversion ratio(P=0.062).Oocysts in the litter were detected in the challenged control diet group and the challenged/essential oil-fed groups at 21 and 35 d,respectively.Superoxide dismutase activity in the serum was elevated(P=0.059)in the salinomycin-fed chickens compared to the challenged controls.Alpha-1-acid glycoprotein was decreased by 28.7%in the salinomycin-fed chickens but increased by 38.1%in the essential oil group compared with the challenged control group.Challenged control group exhibited a significantly higher cooking loss of the thigh meat,compared to the non-challenged control diet group,which was marginally mitigated by dietary supplementation with essential oils.Chickens fed essential oil-added diet had the highest branched-chain fatty acids contents in the cecum.Conclusions:In conclusion,this study shows that oregano and Citrus-based essential oil preparation did not affect growth performance in broiler chickens challenged with the coccidiosis vaccine nor did Eimeria-specific duodenal lesion.However,dietary essential oil preparation lowered oocysts present in litter materials and altered branchedchain fatty acids in cecal digesta.Beneficial findings of the essential oil preparation on volatile fatty acids and oocysts output may warrant further research into assessing its effectiveness and its efficacy in pathogenic fieldisolate Eimeria spp.-induced coccidiosis disease model.