A novel live attenuated vaccine candidate protects chickens against subtype B avian metapneumovirus

Avian metapneumovirus(aMPV) is a highly contagious pathogen that causes acute upper respiratory tract diseases in chickens and turkeys, resulting in serious economic losses. Subtype B aMPV has recently become the dominant epidemic strain in China. We developed an attenuated aMPV subtype B strain by serial passaging in Vero cells and evaluated its safety and efficacy as a vaccine candidate. The safety test showed that after the 30th passage, the LN16-A strain was fully attenuated, as clinical signs of infection and histological lesions were absent after inoculation.The LN16-A strain did not revert to a virulent strain after five serial passages in chickens. The genomic sequence of LN16-A differed from that of the parent wild-type LN16(wtLN16) strain and had nine amino acid mutations. In chickens, a single immunization with LN16-A induced robust humoral and cellular immune responses, including the abundant production of neutralizing antibodies, CD4^(+) T lymphocytes, and the Th1(IFN-γ) and Th2(IL-4 and IL-6)cytokines. We also confirmed that LN16-A provided 100% protection against subtype B aMPV and significantly reduced viral shedding and turbinate inflammation. Our findings suggest that the LN16-A strain is a promising live attenuated vaccine candidate that can prevent infection with subtype B aMPV.

Significant genomic introgression from grey junglefowl(Gallus sonneratii)to domestic chickens(Gallus gallus domesticus)

Background Chicken is one of the most numerous and widely distributed species around the world,and many studies support the multiple ancestral origins of domestic chickens.The research regarding the yellow skin phenotype in domestic chickens(regulated by BCO2)likely originating from the grey junglefowl serves as crucial evidence for demonstrating the multiple origins of chickens.However,beyond the BCO2 gene region,much remains unknown about the introgression from the grey junglefowl into domestic chickens.Therefore,in this study,based on wholegenome data of 149 samples including 4 species of wild junglefowls and 13 local domestic chicken breeds,we explored the introgression events from the grey junglefowl to domestic chickens.Results We successfully detected introgression regions besides BCO2,including two associated with growth trait(IGFBP2 and TKT),one associated with angiogenesis(TIMP3)and two members of the heat shock protein family(HSPB2 and CRYAB).Our findings suggest that the introgression from the grey junglefowl may impact the growth performance of chickens.Furthermore,we revealed introgression events from grey junglefowl at the BCO2 region in multiple domestic chicken breeds,indicating a phenomenon where the yellow skin phenotype likely underwent strong selection and was retained.Additionally,our haplotype analysis shed light on BCO2 introgression event from different sources of grey junglefowl into domestic chickens,possibly suggesting multiple genetic flows between the grey junglefowl and domestic chickens.Conclusions In summary,our findings provide evidences of the grey junglefowl contributing to the genetic diversity of domestic chickens,laying the foundation for a deeper understanding of the genetic composition within domestic chickens,and offering new perspectives on the impact of introgression on domestic chickens.

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%.

Transcriptome-based analysis of key signaling pathways affecting the formation of primordial germ cell in chickens

Bone morphogenetic protein 4(BMP4)can induce the formation of chicken primordial germ cells(PGCs)in vitro;however,its regulatory mechanism in poultry remains unknown.This study aimed to use RNA-seq to analyze PGCs in chicken embryos and iPGCs induced by BMP4 in vitro,clarify the internal regulatory factors of PGCs,analyze the mechanism of the formation of PGCs,and lay a theoretical foundation for the further optimization of PGCs induction systems.Embryonic stem cells(ESCs),PGCs and iPGCs induced by BMP4 in vitro were collected.The transcriptional maps of the three cell types were studied using RNA-seq.The results showed 6,142 genes differentially expressed between PGCs and iPGCs,of which 2,728 were upregulated in iPGCs and 3,414 were downregulated in iPGCs.Compared to that in ESCs,BMP4 was significantly upregulated in PGCs and iPGCs.KEGG results showed that both the TGF-βand Wnt signaling pathways were activated during the formation of PGCs in vitro and in vivo,and the activation was more significant during iPGCs induced by BMP4.The expression of Nodal,an inhibitory factor of TGF-βsignaling,was significantly decreased in PGCs and iPGCs,but was not expressed in iPGCs,which further supports our conclusion.Additionally,the Lysosome and PI3K-AKT signaling pathways were significantly enriched in PGCs and iPGCs,respectively.Further,transmission electron microscopy(TEM)results showed that the number of autolysosomes was significantly higher after the addition of BMP4,which is consistent with the KEGG results.Furthermore,the number of PGCs was significantly reduced after ATG14 was interfered in vivo and in vitro.In conclusion,this study screened out the key signaling pathways during the formation of PGCs,aiming to provide help for enriching the mechanism network regulating PGCs formation in chicken and laying a theoretical foundation for further improving the efficiency of inducing PGCs in vitro.

Retinol is involved in the intestinal regeneration and strengthens the intestinal barrier during refeeding in broiler chickens

Fasting is typically used before feeding metabolizable energy assessment in broilers.Previous studies have shown that fasting cause atrophy of the intestinal villus.Whether fasting affects intestinal permeability during refeeding by altering barrier function and nutrient absorption is of concern.Here,23-d-old broilers were randomly assigned to 5 treatments,fasted for 0,12,24,36,and 48 h,respectively,and then refed for 2 d,to study the impact of different duration of fasting on the intestinal regeneration and barrier function during refeeding.Results showed that the intestinal morphology in fasted birds was recovered in 2 d of refeeding at most.As fasting durations increased,enterocytes per intestinal villus were linearly and quadratically increased(both P<0.05),whereas goblet cells per intestinal villus was linearly decreased(both P<0.05).Besides,the mRNA level of lysozyme was linearly decreased as fasting durations increased during refeeding(both P<0.05),while quadratically increased mucin 2 was observed only after 1 d of refeeding(P<0.05).Linear increase effects were observed for claudin 2 and zonula occludens-1with increased fasting durations after 1 d of refeeding(all P<0.05),and linear and quadratical effects were observed for claudin 2 at 2 d of refeeding(both P<0.05).Besides,we found that intestinal permeability to creatinine,4 and 70 kD dextran were linearly and quadratically decreased with increased fasting durations at 6 h and 1 d of refeeding(all P<0.05).Furthermore,jejunum proteomic from birds refed for 6 h showed that birds fasted for 36 h showed increased antimicrobial peptides and upregulated retinol metabolism when compared to the nonfasted birds(P<0.05).Further study showed that retinyl ester catabolism was inhibited during fasting and enhanced during refeeding.Results of intestinal organoid culture showed that retinol benefits the cell proliferation and enterocyte differentiation.In conclusion,the intestinal permeability to small and large molecules was decreased during refeeding by strengthening the intestinal barrier function,and the activated retinol metabolism during refeeding is involved in the intestinal regeneration and strengthens the intestinal barrier.

Asymmetric expression of CA2 and CA13 linked to calcification in the bilateral mandibular condyles cause crossed beaks in chickens

Crossed beak is a complex mode of inheritance with prevalence ranging from 0.2 to 7.4% in at least 12 chicken strains worldwide.To reveal the intrinsic factors causing crossed beaks,genes expression patterns in bilateral mandibular condyle between affected and normal birds were characterized by RNA sequencing analysis in the present studies.Crossed beak was induced by short length of unilateral mandibular ramus,and a total of 110differentially expressed genes were up-or down-regulated in the affected(short)mandibular condyle side as compared to the normal side.Carbonic anhydrase 2(CA2)and Carbonic anhydrase 13(CA13)were enriched in the carbonate dehydratase activity,and high-expressed in mandibular condyle and osteoblasts(P<0.05).However,both were low-expressed in short mandibular condyle side of affected birds(P<0.05).The carbonate dehydratase inhibitor experiments confirmed that there is positive association between the calcification and carbonic anhydrase isoenzymes.Quantitative analysis with cetylpyridinium chloride showed a decrease in calcification when the cells were transfected with an anti-CA13 shRNA.Our research suggested that CA2 and CA13 are down-calcified in shortside mandibular condyle,and caused mandibular ramus to grow slowly.CA2 and CA13 have the critical role in crossed beaks by regulating calcification of mandibular condyle.

Proteo-transcriptomic profiles reveal key regulatory pathways and functions of LDHA in the ovulation of domestic chickens(Gallus gallus)

Background In poultry, the smooth transition of follicles from the preovulatory-to-postovulatory phase impacts egg production in hens and can benefit the poultry industry. However, the regulatory mechanism underlying follicular ovulation in avians is a complex biological process that remains unclear.Results Critical biochemical events involved in ovulation in domestic chickens(Gallus gallus) were evaluated by transcriptomics, proteomics, and in vitro assays. Comparative transcriptome analyses of the largest preovulatory follicle(F1) and postovulatory follicle(POF1) in continuous laying(CL) and intermittent laying(IL) chickens indicated the greatest difference between CL_F1 and IL_F1, with 950 differentially expressed genes(DEGs), and the smallest difference between CL_POF1 and IL_POF1, with 14 DEGs. Additionally, data-independent acquisition proteomics revealed 252 differentially abundant proteins between CL_F1 and IL_F1. Perivitelline membrane synthesis, steroid biosynthesis, lysosomes, and oxidative phosphorylation were identified as pivotal pathways contributing to ovulation regulation. In particular, the regulation of zona pellucida sperm-binding protein 3, plasminogen activator, cathepsin A, and lactate dehydrogenase A(LDHA) was shown to be essential for ovulation. Furthermore, the inhibition of LDHA decreased cell viability and promoted apoptosis of ovarian follicles in vitro.Conclusions This study reveals several important biochemical events involved in the process of ovulation, as well as crucial role of LDHA. These findings improve our understanding of ovulation and its regulatory mechanisms in avian species.

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.

Major regulatory factors for reproductive performances of female chickens

The reproductive performance of female chickens is critical for determining the efficiency of production and productivity and thus profitability.Studies have shown that the reproductive performance of female chickens is mainly regulated by the feed,hormones,genes,and light conditions.Herein,we review the major factors regulating female chicken reproductive performance and assess the reproductive organs and their functions.In the current review,we highlight how the interconnections of hormones,candidate genes,and photo-stimulation regulate female chicken reproductive hormones and thus regulate the reproductive organ performance.In this regard,the roles of main hormones[gonadotropinreleasing hormone(GnRH)and genes(GnRH-Ⅰ)]in regulating sexual maturation and ovarian development and maintenance by influencing the survival and function of follicular granulosa cells were also reviewed.In addition,the current review also highlights how feeding female chickens with diets and artificial light-emitting diodes(LEDs)support the effective functioning of their reproductive capacity through the stimulation of sexual maturity at an appropriate age and regeneration of aged reproductive organs.

Study on Outer Membrane Protein Patterns of Escherichia coli O38,O53 and O75 Isolated from Chickens

[Objective] This study aimed to investigate the outer membrane protein （OMP） patterns of Escherichia coli 038, 053 and 075 isolates from chickens. [Method] Eight pathogenic E. coil isolates with various serotypes were used as experimental materials to extract OMP by using supersonic schizolysis method and Sarcosyl. After SDS-PAGE electrophoresis, OMP patterns of the extracted products were determined based on the OMP model diagram. [Result] OMP of eight E. coil isolates with three serotypes were divided into three patterns, to be specific, 2 075 isolates respectively belonged to OMP-I and OMP-II pattern, 1 053 isolate belonged to OMP-II pattern, and 5 038 isolates belonged to OMP-I and OMP-III pattern. [Conclusion] Experimental results showed that E. coli isolates with the same serotype may belong to completely different OMP patterns, while serologically unrelated isolates may belong to the same OMP pattern. OMP of E. coil isolates with the same serotype may generate genetic differentiation; in addition, OMP of E. coli isolates with different serotypes may have different genetic correlation.

Effects of Bacillus coagulans supplementation on the growth performance and gut health of broiler chickens with Clostridium perfringens-induced necrotic enteritis

Background: The poultry industry is in need of effective antibiotic alternatives to control outbreaks of necrotic enteritis(NE) due to Clostridium perfringens.Methods: This study was conducted to investigate the effects of feeding Bacil us coagulans on the growth performance and gut health of broiler chickens with C. perfringens-induced NE. Two hundred and forty 1-day-old broiler chicks were randomly assigned to a 2 × 2 factorial arrangement with two dietary B. coagulans levels(0 or 4 × 109 CFU/kg of diet) and two disease chal enge statuses(control or NE chal enged).Results: NE-induced reduction in body weight gain was relieved by the addition of B. coagulans into broiler diets compared with the NE-infected birds. NE infection damaged intestinal morphological structure, promoted intestinal C.perfringens growth and liver invasion, and enhanced anti-C. perfringens specific sI gA concentrations in the gut and specific IgG levels in serum compared with the uninfected birds. NE infection significantly(P < 0.05) decreased mucin-2(at 14 d post-infection(DPI), tol-like receptor 2(TLR2, at 7 and 14 DPI), TLR4(at 7 and 14 DPI), tumor necrosis factor super family15(TNFSF15, at 7 and 14 DPI), lysozyme(LYZ, at 14 DPI) and fowlicidin-2(at 7 and 14 DPI) mR NA levels, whereas it dramatical y(P = 0.001) increased IFN-γ mR NA levels at 7 DPI. However, chal enged birds fed diets supplemented with B.coagulans showed a significant(P < 0.01) decrease in gut lesion scores, decreased C. perfringens numbers in the cecum and liver, and an increase in fowlicidin-2 mR NA levels in compared with the uninfected birds. In addition, compared with the non-supplemented group, dietary inclusion of B. coagulans improved intestinal barrier structure, further increased specific sI gA levels and alkaline phosphatase(IAP) activity in the jejunum, enhanced the expression of jejunum lysozyme mR NA, and inhibited the growth, colonization, and invasion of C. perfringens; in contrast, it reduced serum-specific IgG concentrations and jejunum IFN-γ mR NA levels.Conclusion: These results indicated that dietary B. coagulans supplementation appeared to be effective in preventing the occurrence and reducing the severity of C. perfringens-induced NE in broiler chickens.

Influencing Factors and Variation Trends of Muscle pH in Chickens after Slaughter

[Objective] This study aimed to investigate influencing factors and variation trends of muscle p H in chickens after slaughter. [Method] Beijing You chickens and Lohmann brown chickens were reared under the same conditions and slaughtered at the age of 8, 16 and 20 weeks, respectively. At different time（15 min, 1, 4, 8,12, 24 and 48 h） after slaughter, p H value of breast muscles and leg muscles was measured. [Result] The results showed that muscle position, post-slaughter time,gender and week age were important factors influencing p H value, but the breed had little effect. Within 1 h after slaughter, p H value of breast muscles and leg muscles declined rapidly, which reached the minimum level at 12 h after slaughter,and then increased slowly. In addition, p H value of leg muscles exhibited higher decline and increase rates than that of breast muscles to some extent. [Conclusion]This study provided basis for breeding of chickens with excellent meat quality.

Characterization of Genetic Polymorphism of Novel MHC B-LBⅡ Alleles in Chinese Indigenous Chickens

Genetic polymorphism of the major histocompatibility complex （MHC） B-LBⅡ gene was studied by amplification of exon 2 using PCR, followed by cloning and DNA sequencing in eight indigenous Chinese chicken populations. To reveal the genetic variation of the B-LB Ⅱ gene, 37 types of patterns detected by PCR-SSCP were investigated first, which would be used to screen novel B-LB Ⅱsequences within the breeds. The types of PCR-SSCP patterns and final sequencing allowed for the identification of 31 novel MHC B-LBⅡ alleles from 30 unrelated individuals of Chinese chickens that were sampled. These are the first designators for the alleles of chicken MHC B-LBⅡ gene based on the rule of assignment for novel mammalian alleles. Sequence alignment of the 31 B-LB Ⅱ alleles revealed a total of 68 variable sites in the fragment of exon 2, of which 51 parsimony informative and 17 singleton variable sites were observed. Among the polymorphic sites, the nucleotide substitutions in the first and second positions of the codons accounted for 36.76% and 35.29%, respectively. The sequence similarities between the alleles were estimated to be 90.6%-99.5%. The relative frequencies of synonymous and nonsynonymous nucleotide substitutions within the region were 2.92%±0.94% and 14.64%±2.67%, respectively. These results indicated that the genetic variation within exon 2 appeared to have largely arisen by gene recombination and balancing selection. Alignment of the deduced amino acid sequences of the β1 domain coded by exon 2 revealed 6 synonymous mutations and 27 nonsynonymous substitutions at the 33 disparate sites. In particular, the nonsynonymous substitutions at the putative peptide-binding sites are considered to be associated with immunological specificity of MHC B-LB Ⅱ molecule in Chinese native chickens. These results can provide a molecular biological basis for the study of disease resistance in chicken breeding.

Black soldier fly defatted meal as a dietary protein source for broiler chickens:Effects on growth performance,blood traits,gut morphology and histological features

Background: The present study has evaluated the effects of different inclusion levels of a partially defatted black soldier fly(Hermetia illucens L.; HI) larva meal on the growth performance, blood parameters and gut morphology of broiler chickens. A total of 256 male broiler chickens(Ross 308) were reared from d 1 to d 35 and assigned to4 dietary treatments(8 replicates/treatment and 8 birds/replicate). HI larva meal was included at increasing levels(0, 5%, 10% and 15%; HI0, HI5, HI10 and HI15, respectively) in isonitrogenous and isoenergetic diets formulated for 3 feeding phases: starter(1–10 d), growing(10–24 d) and finisher(24–35 d). Two birds per pen were slaughtered at d 35 and morphometric investigations and histopathological alterations were performed.Results: The live weight(LW) showed linear and quadratic responses to increasing HI larva meal(maximum for HI10 group). Average daily gain(ADG) showed a linear and quadratic responses to HI meal(maximum for HI10 group) during starter and growing periods. A linear decrease was observed for ADG during the finisher period. The daily feed intake(DFI) showed a linear and quadratic effect during the starter period(maximum for HI10 group).Linear and quadratic responses were observed for the feed conversion ratio(FCR) in the growing period and for the whole period of the experiment. The FCR showed a linear response in the finisher period(maximum for HI15).No significant effects were observed for the blood and serum parameters, except for the phosphorus concentration,which showed linear and quadratic responses as well as glutathione peroxidase(GPx) activity, the latter of which showed a linear response. The HI15 birds showed a lower villus height, a higher crypt depth and a lower villus height-to-crypt depth ratio than the other groups.Conclusions: Increasing levels of dietary HI meal inclusion in male broiler chickens may improve the LW and DFI during the starter period, but may also negatively affect the FCR and gut morphology, thus suggesting that low levels may be more suitable. However, no significant effects on the haematochemical parameters or histological findings were observed in relation to HI meal utilization.

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.

Influence of starch sources and dietary protein levels on intestinal functionality and intestinal mucosal amino acids catabolism in broiler chickens

Background: There is growing interest in carbohydrate and protein nutrition to enhance the efficiency of animal production.Reduced-crude protein diets depress environmental pollution and feeding cost,but the challenge to their adoption is maintaining digestive function and growth performance of birds.The present study was conducted to evaluate the influence of different dietary starch sources and protein levels on intestinal functionality and mucosal amino acid catabolism.Methods: Six dietary treatments,based on maize and soybean meal,were offered to 360 AA+male chicks from 6 to 35 d post-hatch as a 3 × 2 factorial array.Either waxy rice or amylose was added to a conventional maize-soy diet to provide three sources of starch with different digestion rates and relatively high and low dietary protein levels.Growth performance,parameters of intestinal functionality and concentrations of free amino acid in the portal circulation were determined.Results: In the grower phase,starch source influenced(P < 0.02) weight gain as diets containing amylose supported significantly higher weight gains than waxy rice.Significant increase of ileal ATP concentrations and Na^+/K^+-ATPase activity were found in amylose treatment.Also,amylose decreased BrdU positive cell numbers and down-regulated m RNA expression for CASP-3.GOT activity in the ileum was higher(P < 0.01) in birds offered low protein diets and there was a trend(P = 0.057) for waxy rice as a starch source to increase ileal GOT activities.There was a significant influence on the concentration of seventeen amino acids in the portal circulation with tryptophan the one exception.Waxy rice as a starch source generated 13.6% and 22.4% numerically higher concentrations of non-essential amino acids than maize and amylose,respectively.Conclusions: Amino acid catabolism in the gut mucosa is subject to nutritional regulation.Given that amino acids can be spared from catabolism in the gut mucosa by supplementation of amylose,it follows their post-enteral availability would be improved and intestinal energy would be derived more efficiently from glucose.

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.

Intestinal toxicity of deoxynivalenol is limited by supplementation with Lactobacillus plantarum JM113 and consequentially altered gut microbiota in broiler chickens

Background: Limited research has focused on the effect of Lactobacillus on the intestinal toxicity of deoxynivalenol(DON).The present study was conducted to investigate the role of Lactobacillus plantarum(L.plantarum) JM113 in protecting against the intestinal toxicity caused by DON.Methods: A total of 144 one-day-old healthy Arbor Acres broilers were randomly distributed into 3 treatments,including the CON(basal diet),the DON(extra 10 mg/kg deoxynivalenol),and the DL(extra 1 × 109 CFU/kg L.plantarum JM113 based on DON group) treatments.The growth performance,organ indexes,intestinal morphology,pancreatic digestive enzymes,intestinal secreted immunoglobulin A(sIgA),jejunal transcriptome,and intestinal microbiota were evaluated.Results: Compared with the CON and DL groups,the DON supplementation altered intestinal morphology,especially in duodenum and jejunum,where villi were shorter and crypts were deeper(P < 0.05).Meanwhile,the significantly decreased mRNA expression of jejunal claudin-1 and occludin(P < 0.05),ileal rBAT and jejunal GLUT1 of 21-day-old broilers(P < 0.05),as well as duodenal PepT1 and ileal rBAT of 42-day-old broilers were identified in the DON group.Moreover,supplementation with L.plantarum JM113 could increase duodenal expression of IL-10 and IL-12 of 21-dayold broilers,ileal s IgA of 42-day-old broilers,and the bursa of Fabricius index of 21-day-old broilers.Further jejunal transcriptome proved that the genes related to the intestinal absorption and metabolism were significantly reduced in the DON group but a significant increase when supplemented with extra L.plantarum JM113.Furthermore,the bacteria related to nutrient utilization,including the Proteobacteria,Escherichia,Cc-115(P < 0.05),Lactobacillus and Prevotella(P < 0.1) were all decreased in the DON group.By contrast,supplementation with L.plantarum JM113 increased the relative abundance of beneficial bacterium,including the Bacteroidetes,Roseburia,Anaerofustis,Anaerostipe,and Ruminococcus bromi(P < 0.05).Specifically,the increased abundance of bacteria in the DL group could be proved by the significantly increased caecal content of propionic acid,n-Butyric acid,and total short-chain fatty acid.Conclusions: L.plantarum JM113 enhanced the digestion,absorption,and metabolic functions of the gut when challenged with DON by reducing the injury to intestinal barriers and by increasing the abundance of beneficial bacterium.

Protective efficacy of an H5/H7 trivalent inactivated vaccine(H5-Re13,H5-Re14, and H7-Re4 strains) in chickens, ducks, and geese against newly detected H5N1, H5N6, H5N8, and H7N9 viruses

Some H5 viruses isolated in poultry or wild birds between 2020 and 2021 were found to be antigenically different from the vaccine strains(H5-Re11 and H5-Re12) used in China. In this study, we generated three new recombinant vaccine seed viruses by using reverse genetics and used them for vaccine production. The vaccine strain H5-Re13 contains the hemagglutinin(HA) and neuraminidase(NA) genes of an H5 N6 virus that bears the clade 2.3.4.4 h HA gene, H5-Re14 contains the HA and NA genes of an H5 N8 virus that bears the clade 2.3.4.4 b HA gene, and H7-Re4 contains the HA and NA genes of H7 N9 virus detected in 2021. We evaluated the protective efficacy of the novel H5/H7 trivalent inactivated vaccine in chickens, ducks, and geese. The inactivated vaccine was immunogenic and induced substantial antibody responses in the birds tested. Three weeks after vaccination, chickens were challenged with five different viruses detected in 2020 and 2021: three viruses(an H5 N1 virus, an H5 N6 virus, and an H5 N8 virus) bearing the clade 2.3.4.4 b HA gene, an H5 N6 virus bearing the clade 2.3.4.4 h HA gene, and an H7 N9 virus. All of the control birds shed high titers of virus and died within 4 days post-challenge, whereas the vaccinated chickens were completely protected from these viruses. Similar protective efficacy against H5 viruses bearing the clade 2.3.4.4 h or 2.3.4.4 b HA gene was observed in ducks and geese. Our study indicates that the newly updated H5/H7 vaccine can provide solid protection against the H5 and H7 N9 viruses that are currently circulating in nature.