Multi-omics analysis reveals the molecular regulatory network underlying the prevention of Lactiplantibacillus plantarum against LPS-induced salpingitis in laying hens

Background Salpingitis is one of the common diseases in laying hen production, which greatly decreases the economic outcome of laying hen farming. Lactiplantibacillus plantarum was effective in preventing local or systemic inflammation, however rare studies were reported on its prevention against salpingitis. This study aimed to investigate the preventive molecular regulatory network of microencapsulated Lactiplantibacillus plantarum(MLP) against salpingitis through multi-omics analysis, including microbiome, transcriptome and metabolome analyses.Results The results revealed that supplementation of MLP in diet significantly alleviated the inflammation and atrophy of uterus caused by lipopolysaccharide(LPS) in hens(P < 0.05). The concentrations of plasma IL-2 and IL-10 in hens of MLP-LPS group were higher than those in hens of LPS-stimulation group(CN-LPS group)(P < 0.05). The expression levels of TLR2, MYD88, NF-κB, COX2, and TNF-α were significantly decreased in the hens fed diet supplemented with MLP and suffered with LPS stimulation(MLP-LPS group) compared with those in the hens of CN-LPS group(P < 0.05). Differentially expressed genes(DEGs) induced by MLP were involved in inflammation, reproduction, and calcium ion transport. At the genus level, the MLP supplementation significantly increased the abundance of Phascolarctobacterium, whereas decreased the abundance of Candidatus_Saccharimonas in LPS challenged hens(P < 0.05). The metabolites altered by dietary supplementation with MLP were mainly involved in galactose, uronic acid, histidine, pyruvate and primary bile acid metabolism. Dietary supplementation with MLP inversely regulates LPSinduced differential metabolites such as Lyso PA(24:0/0:0)(P < 0.05).Conclusions In summary, dietary supplementation with microencapsulated Lactiplantibacillus plantarum prevented salpingitis by modulating the abundances of Candidatus_Saccharimonas, Phascolarctobacterium, Ruminococcus_torques_group and Eubacterium_hallii_group while downregulating the levels of plasma metabolites, p-tolyl sulfate, o-cresol and N-acetylhistamine and upregulating S-lactoylglutathione, simultaneously increasing the expressions of CPNE4, CNTN3 and ACAN genes in the uterus, and ultimately inhibiting oviducal inflammation.

Effects of different energy levels in low-protein diet on liver lipid metabolism in the late-phase laying hens through the gut-liver axis

Background The energy/protein imbalance in a low-protein diet induces lipid metabolism disorders in late-phase laying hens.Reducing energy levels in the low-protein diet to adjust the energy-to-protein ratio may improve fat deposition,but this also decreases the laying performance of hens.This study investigated the mechanism by which different energy levels in the low-protein diet influences liver lipid metabolism in late-phase laying hens through the enterohepatic axis to guide feed optimization and nutrition strategies.A total of 288 laying hens were randomly allocated to the normal-energy and normal-protein diet group(positive control:CK)or 1 of 3 groups:lowenergy and low-protein diet(LL),normal-energy and low-protein diet(NL),and high-energy and low-protein diet(HL)groups.The energy-to-protein ratios of the CK,LL,NL,and HL diets were 0.67,0.74,0.77,and 0.80,respectively.Results Compared with the CK group,egg quality deteriorated with increasing energy intake in late-phase laying hens fed low-protein diet.Hens fed LL,NL,and HL diets had significantly higher triglyceride,total cholesterol,acetylCo A carboxylase,and fatty acid synthase levels,but significantly lower hepatic lipase levels compared with the CK group.Liver transcriptome sequencing revealed that genes involved in fatty acid beta-oxidation(ACOX1,HADHA,EHHADH,and ACAA1)were downregulated,whereas genes related to fatty acid synthesis(SCD,FASN,and ACACA)were upregulated in LL group compared with the CK group.Comparison of the cecal microbiome showed that in hens fed an LL diet,Lactobacillus and Desulfovibrio were enriched,whereas riboflavin metabolism was suppressed.Cecal metabolites that were most significantly affected by the LL diet included several vitamins,such as riboflavin(vitamin B2),pantethine(vitamin B5 derivative),pyridoxine(vitamin B6),and 4-pyridoxic acid.Conclusion A lipid metabolism disorder due to deficiencies of vitamin B2 and pantethine originating from the metabolism of the cecal microbiome may be the underlying reason for fat accumulation in the liver of late-phase laying hens fed an LL diet.Based on the present study,we propose that targeting vitamin B2 and pantethine(vitamin B5 derivative)might be an effective strategy for improving lipid metabolism in late-phase laying hens fed a low-protein diet.

Rosemary extract improves egg quality by altering gut barrier function,intestinal microbiota and oviductal gene expressions in late-phase laying hens

Background Rosemary extract(RE)has been reported to exert antioxidant property.However,the application of RE in late-phase laying hens on egg quality,intestinal barrier and microbiota,and oviductal function has not been systematically studied.This study was investigated to detect the potential effects of RE on performance,egg quality,serum parameters,intestinal heath,cecal microbiota and metabolism,and oviductal gene expressions in late-phase laying hens.A total of 21065-week-old“Jing Tint 6”laying hens were randomly allocated into five treatments with six replicates and seven birds per replicate and fed basal diet(CON)or basal diet supplemented with chlortetracycline at 50 mg/kg(CTC)or RE at 50 mg/kg(RE50),100 mg/kg(RE100),and 200 mg/kg(RE200).Results Our results showed that RE200 improved(P<0.05)Haugh unit and n-6/n-3 of egg yolk,serum superoxide dismutase(SOD)compared with CON.No significant differences were observed for Haugh unit and n-6/n-3 of egg yolk among CTC,RE50,RE100 and RE200 groups.Compared with CTC and RE50 groups,RE200 increased serum SOD activity on d 28 and 56.Compared with CON,RE supplementation decreased(P<0.05)total cholesterol(TC)level.CTC,RE100 and RE200 decreased(P<0.05)serum interleukin-6(IL-6)content compared with CON.CTC and RE200 increased jejunal m RNA expression of ZO-1 and Occludin compared with CON.The biomarkers of cecal microbiota and metabolite induced by RE 200,including Firmicutes,Eisenbergiella,Paraprevotella,Papillibacter,and butyrate,were closely associated with Haugh unit,n-6/n-3,SOD,IL-6,and TC.PICRUSt2 analysis indicated that RE altered carbohydrate and amino acid metabolism of cecal microbiota and increased butyrate synthesizing enzymes,including 3-oxoacid Co A-transferase and butyrate-acetoacetate Co A-transferase.Moreover,transcriptomic analysis revealed that RE200 improved gene expressions and functional pathways related to immunity and albumen formation in the oviductal magnum.Conclusions Dietary supplementation with 200 mg/kg RE could increase egg quality of late-phase laying hens via modulating intestinal barrier,cecal microbiota and metabolism,and oviductal function.Overall,RE could be used as a promising feed additive to improve egg quality of laying hens at late stage of production.

Prevotella and succinate treatments altered gut microbiota,increased laying performance,and suppressed hepatic lipid accumulation in laying hens

Background This work aimed to investigate the potential benefits of administering Prevotella and its primary metabolite succinate on performance,hepatic lipid accumulation and gut microbiota in laying hens.Results One hundred and fifty 58-week-old Hyline Brown laying hens,with laying rate below 80%and plasma triglyceride(TG)exceeding 5 mmol/L,were used in this study.The hens were randomly allocated into 5 groups and subjected to one of the following treatments:fed with a basal diet(negative control,NC),oral gavage of 3 mL/hen saline every other day(positive control,PC),gavage of 3 mL/hen Prevotella melaninogenica(10^(7)CFU/mL,PM)or 3 mL/hen Prevotella copri(10^(7)CFU/mL,P.copri)every other day,and basal diet supplemented with 0.25%sodium succinate(Succinate).The results showed that PM and P.copri treatments significantly improved laying rate compared to the PC(P<0.05).The amount of lipid droplet was notably decreased by PM,P.copri,and Succinate treatments at week 4 and decreased by P.copri at week 8(P<0.05).Correspondingly,the plasma TG level in Succinate group was lower than that of PC(P<0.05).Hepatic TG content,however,was not significantly influenced at week 4 and 8(P>0.05).PM treatment increased(P<0.05)the mRNA levels of genes PGC-1βand APB-5B at week 4,and ACC and CPT-1 at week 8.The results indicated enhanced antioxidant activities at week 8,as evidenced by reduced hepatic malondialdehyde(MDA)level and improved antioxidant enzymes activities in PM and Succinate groups(P<0.05).Supplementing with Prevotella or succinate can alter the cecal microbiota.Specifically,the abundance of Prevotella in the Succinate group was significantly higher than that in the other 4 groups at the family and genus levels(P<0.05).Conclusions Oral intake of Prevotella and dietary supplementation of succinate can ameliorate lipid metabolism of laying hens.The beneficial effect of Prevotella is consistent across different species.The finding highlights that succinate,the primary metabolite of Prevotella,represents a more feasible feed additive for alleviating fatty liver in laying hens.

Decreased eggshell strength caused by impairment of uterine calcium transport coincide with higher bone minerals and quality in aged laying hens

Background Deteriorations in eggshell and bone quality are major challenges in aged laying hens.This study compared the differences of eggshell quality,bone parameters and their correlations as well as uterine physiologi-cal characteristics and the bone remodeling processes of hens laying eggs of different eggshell breaking strength to explore the mechanism of eggshell and bone quality reduction and their interaction.A total of 24074-week-old Hy-line Brown laying hens were selected and allocated to a high(HBS,44.83±1.31 N)or low(LBS,24.43±0.57 N)eggshell breaking strength group.Results A decreased thickness,weight and weight ratio of eggshells were observed in the LBS,accompanied with ultrastructural deterioration and total Ca reduction.Bone quality was negatively correlated with eggshell quality,marked with enhanced structures and increased components in the LBS.In the LBS,the mammillary knobs and effective layer grew slowly.At the initiation stage of eggshell calcification,a total of 130 differentially expressed genes(DEGs,122 upregulated and 8 downregulated)were identified in the uterus of hens in the LBS relative to those in the HBS.These DEGs were relevant to apoptosis due to the cellular Ca overload.Higher values of p62 protein level,caspase-8 activity,Bax protein expression and lower values of Bcl protein expression and Bcl/Bax ratio were seen in the LBS.TUNEL assay and hematoxylin-eosin staining showed a significant increase in TUNEL-positive cells and tissue damages in the uterus of the LBS.Although few DEGs were identified at the growth stage,similar uterine tissue damages were also observed in the LBS.The expressions of runt-related transcription factor 2 and osteocal-cin were upregulated in humeri of the LBS.Enlarged diameter and more structural damages of endocortical bones and decreased ash were observed in femurs of the HBS.Conclusion The lower eggshell breaking strength may be attributed to a declined Ca transport due to uterine tissue damages,which could affect eggshell calcification and lead to a weak ultrastructure.Impaired uterine Ca transport may result in reduced femoral bone resorption and increased humeral bone formation to maintain a higher mineral and bone quality in the LBS.

Coated sodium butyrate ameliorates high‑energy and low‑protein diet induced hepatic dysfunction via modulating mitochondrial dynamics, autophagy and apoptosis in laying hens

Background Fatty liver hemorrhagic syndrome(FLHS),a fatty liver disease in laying hens,poses a grave threat to the layer industry,stemming from its ability to trigger an alarming plummet in egg production and usher in acute mortality among laying hens.Increasing evidence suggests that the onset and progression of fatty liver was closely related to mitochondria dysfunction.Sodium butyrate was demonstrated to modulate hepatic lipid metabolism,alle-viate oxidative stress and improve mitochondrial dysfunction in vitro and mice models.Nevertheless,there is limited existing research on coated sodium butyrate(CSB)to prevent FLHS in laying hens,and whether and how CSB exerts the anti-FLHS effect still needs to be explored.In this experiment,the FLHS model was induced by administering a high-energy low-protein(HELP)diet in laying hens.The objective was to investigate the effects of CSB on alleviating FLHS with a focus on the role of CSB in modulating mitochondrial function.Methods A total of 288 healthy 28-week-old Huafeng laying hens were arbitrarily allocated into 4 groups with 6 replicates each,namely,the CON group(normal diet),HELP group(HELP diet),CH500 group(500 mg/kg CSB added to HELP diet)and CH750 group(750 mg/kg CSB added to HELP diet).The duration of the trial encompassed a period of 10 weeks.Results The result revealed that CSB ameliorated the HELP-induced FLHS by improving hepatic steatosis and patho-logical damage,reducing the gene levels of fatty acid synthesis,and promoting the mRNA levels of key enzymes of fatty acid catabolism.CSB reduced oxidative stress induced by the HELP diet,upregulated the activity of GSH-Px and SOD,and decreased the content of MDA and ROS.CSB also mitigated the HELP diet-induced inflammatory response by blocking TNF-α,IL-1β,and F4/80.In addition,dietary CSB supplementation attenuated HELP-induced activation of the mitochondrial unfolded protein response(UPRmt),mitochondrial damage,and decline of ATPase activity.HELP diet decreased the autophagosome formation,and downregulated LC3B but upregulated p62 protein expression,which CSB administration reversed.CSB reduced HELP-induced apoptosis,as indicated by decreases in the Bax/Bcl-2,Caspase-9,Caspase-3,and Cyt C expression levels.Conclusions Dietary CSB could ameliorate HELP diet-induced hepatic dysfunction via modulating mitochondrial dynamics,autophagy,and apoptosis in laying hens.Consequently,CSB,as a feed additive,exhibited the capacity to prevent FLHS by modulating autophagy and lipid metabolism.

Lactiplantibacillus plantarum FRT4 attenuates high-energy low-protein dietinduced fatty liver hemorrhage syndrome in laying hens through regulating gut-liver axis

Background Fatty liver hemorrhage syndrome(FLHS)becomes one of the most major factors resulting in the laying hen death for caged egg production.This study aimed to investigate the therapeutic effects of Lactiplantibacillus plan-tarum(Lp.plantarum)FRT4 on FLHS model in laying hen with a focus on liver lipid metabolism,and gut microbiota.Results The FLHS model of laying hens was established by feeding a high-energy low-protein(HELP)diet,and the treatment groups were fed a HELP diet supplemented with differential proportions of Lp.plantarum FRT4.The results indicated that Lp.plantarum FRT4 increased laying rate,and reduced the liver lipid accumulation by regulating lipid metabolism(lipid synthesis and transport)and improving the gut microbiota composition.Moreover,Lp.plan-tarum FRT4 regulated the liver glycerophospholipid metabolism.Meanwhile,“gut-liver”axis analysis showed that there was a correlation between gut microbiota and lipid metabolites.Conclusions The results indicated that Lp.plantarum FRT4 improved the laying performance and alleviated FLHS in HELP diet-induced laying hens through regulating“gut-liver”axis.Our findings reveal that glycerophospholipid metabolism could be the underlying mechanism for the anti-FLHS effect of Lp.plantarum FRT4 and for future use of Lp.plantarum FRT4 as an excellent additive for the prevention and mitigation of FLHS in laying hens.

Effect of dietary supplementation with discardednutrient medium of Cordyceps militaris on layingperformance and diversity of intestinal flora in laying hens

The aim of the experiment was to explore the feasibility of discarded nutrient medium of Cordyceps militaris as feed in the production of laying hens. 100 g/kg, 200 g/kg, 300 g/kg of discarded nutrient medium of Cordyceps militaris were added to the basal diet of laying hens. The results showed that the optimal addition of discarded nutrient medium of Cordyceps militaris in the diet of laying hens was 10%. According to the results of measuring the conventional indicators of eggs, the weight of eggs produced by laying hens fed with discarded nutrient medium of Cordyceps militaris was higher than that of laying hens fed with ordinary laying hens. The content of interleukin-1(IL-1) in the experimental group was significantly higher than that in the control group, and the concentration of IL-1 increased by 141.5 pg/mL, which indicated that the application of discarded nutrient medium of Cordyceps militaris effectively improved the immunity of laying hens. The high-throughput analysis of the intestinal contents of the two groups of laying hens showed that the microbial population abundance of the intestinal tract of the experimental group was greater than that of control group, and the application of discarded nutrient medium of Cordyceps militaris increased the diversity of bacteria in the intestinal tract of laying hens. In addition, the sensitivity of some pathogenic bacteria in the intestinal tract of chickens to drugs was also increased, thereby reducing the use of antibiotics. The secondary utilization of discarded nutrient medium of Cordyceps militaris has great development and utilization prospects, which provided a scientific reference and basic theoretical basis for the development of discarded nutrient medium of Cordyceps militaris as feed in the production of laying hens.

Dietary supplementation of Macleaya cordata extract and Bacillus in combination improve laying performance by regulating reproductive hormones,intestinal microbiota and barrier function of laying hens

Background:This study aimed to investigate whether the combination of Macleaya cordata extract(MCE)and Bacil-lus could improve the laying performance and health of laying hens better.Methods:A total of 36029-week-old Jingbai laying hens were randomly divided into 4 treatments:control group(basal diet),MCE group(basal diet+MCE),Probiotics Bacillus Compound(PBC)group(basal diet+compound Bacil-lus),MCE+PBC group(basal diet+MCE+compound Bacillus).The feeding experiment lasted for 42 d.Results:The results showed that the laying rate and the average daily egg mass in the MCE+PBC group were significantly higher than those in the control group(P<0.05)and better than the MCE and PBC group.Combina-tion of MCE and Bacillus significantly increased the content of follicle-stimulating hormone(FSH)in the serum and up-regulated the expression of related hormone receptor gene(estrogen receptor-β,FSHR and luteinizing hormone/choriogonadotropin receptor)in the ovary of laying hens(P<0.05).In the MCE+PBC group,the mRNA expressions of zonula occluden-1,Occludin and mucin-2 in jejunum was increased and the intestinal epithelial barrier detected by transmission electron microscopy was enhanced compared with the control group(P<0.05).In addition,compared with the control group,combination of MCE and Bacillus significantly increased the total antioxidant capacity and catalase activity(P<0.05),and down-regulated the mRNA expressions of inflammation-related genes(interleukin-1βand tumor necrosis factor-α)as well as apoptosis-related genes(Caspase 3,Caspase 8 and P53)(P<0.05).The concen-tration of acetic acid and butyric acid in the cecum content of laying hens in the MCE+PBC group was significantly increased compared with the control group(P<0.05).Conclusions:Collectively,dietary supplementation of 600μg/kg MCE and 5×108 CFU/kg compound Bacillus can improve laying performance by improving microbiota to enhance antioxidant capacity and intestinal barrier,regulate reproductive hormones and the concentration of cecal short-chain fatty acids of laying hens,and the combined effect of MCE and Bacillus is better than that of single supplementation.

Adjusting phosphate feeding regimen according to daily rhythm increases eggshell quality via enhancing medullary bone remodeling in laying hens

Background Body phosphorus metabolism exhibits a circadian rhythm over the 24-h daily cycle.The egg laying behavior makes laying hens a very special model for investigating phosphorus circadian rhythms.There is lack of information about the impact of adjusting phosphate feeding regimen according to daily rhythm on the phosphorus homeostasis and bone remodeling of laying hens.Methods and results Two experiments were conducted.In Exp.1,Hy-Line Brown laying hens(n=45)were sampled according the oviposition cycle(at 0,6,12,and 18 h post-oviposition,and at the next oviposition,respectively;n=9 at each time point).Diurnal rhythms of body calcium/phosphorus ingestions and excretions,serum calcium/phosphorus levels,oviduct uterus calcium transporter expressions,and medullary bone(MB)remodeling were illustrated.In Exp.2,two diets with different phosphorus levels(0.32%and 0.14%non-phytate phosphorus(NPP),respectively)were alternately presented to the laying hens.Briefly,four phosphorus feeding regimens in total(each included 6 replicates of 5 hens):(1)fed 0.32%NPP at both 09:00 and 17:00;(2)fed 0.32%NPP at 09:00 and 0.14%NPP at 17:00;(3)fed 0.14%NPP at 09:00 and 0.32%NPP at 17:00;(4)fed 0.14%NPP at both 09:00 and 17:00.As a result,the regimen fed 0.14%NPP at 09:00 and 0.32%NPP at 17:00,which was designed to strengthen intrinsic phosphate circadian rhythms according to the findings in Exp.1,enhanced(P<0.05)MB remodeling(indicated by histological images,serum markers and bone mineralization gene expressions),elevated(P<0.05)oviduct uterus calcium transportation(indicated by transient receptor potential vanilloid 6 protein expression),and subsequently increased(P<0.05)eggshell thickness,eggshell strength,egg specific gravity and eggshell index in laying hens.Conclusions These results underscore the importance of manipulating the sequence of daily phosphorus ingestion,instead of simply controlling dietary phosphate concentrations,in modifying the bone remodeling process.Body phosphorus rhythms will need to be maintained during the daily eggshell calcification cycle.

Chronic heat stress induces renal fibrosis and mitochondrial dysfunction in laying hens

Background Heat stress in laying hens negatively affects egg production and shell quality by disrupting the homeo-stasis of plasma calcium and phosphorus levels.Although the kidney plays an important role in calcium and phos-phorus homeostasis,evidence regarding the effect of heat stress on renal injury in laying hens is yet to be elucidated.Therefore,the aim of this study was to evaluate the effects of chronic heat stress on renal damage in hens during laying periods.Methods A total of 16 white-leghorn laying hens(32 weeks old)were randomly assigned to two groups(n=8).One group was exposed to chronic heat stress(33°C for 4 weeks),whereas the other group was maintained at 24°C.Results Chronic heat exposure significantly increased plasma creatinine and decreased plasma albumin levels(P<0.05).Heat exposure also increased renal fibrosis and the transcription levels of fibrosis-related genes(COLA1A1,αSMA,and TGF-β)in the kidney.These results suggest that renal failure and fibrosis were induced by chronic heat exposure in laying hens.In addition,chronic heat exposure decreased ATP levels and mitochondrial DNA copy number(mtDNA-CN)in renal tissue,suggesting that renal mitochondrial dysfunction occurs under conditions of heat stress.Damaged mitochondria leak mtDNAs into the cytosol and mtDNA leakage may activate the cyclic GMP-AMP synthase(cGAS)stimulator of interferon genes(STING)signaling pathway.Our results showed that chronic heat exposure activated the cGAS-STING pathway as indicated by increased expression of MDA5,STING,IRF7,MAVS,and NF-κB levels.Furthermore,the expression of pro-inflammatory cytokines(IL-12)and chemokines(CCL4 and CCL20)was upregulated in heat-stressed hens.Conclusions These results suggest that chronic heat exposure induces renal fibrosis and mitochondrial damage in laying hens.Mitochondrial damage by heat stress may activate the mtDNA-cGAS-STING signaling and cause subse-quent inflammation,which contributes to the progression of renal fibrosis and dysfunction.

Honoring a legend: Celebrating the life and legacy of Professor Heng Li

Professor Heng Li(李恒,March 9,1929-January 12,2023)was a legendary plant taxonomist and phytogeographer at the Kunming Institute of Botany,Chinese Academy of Sciences(Fig.1).She made important contributions to our understanding of China's flora,including the biodiversity and biogeography of Dulong Valley and the Gaoligong Mountains,as well as the vegetation of plateau lakes and wetlands in Southwest China.Professor Heng Li received her Bachelor's degree in Russian from Beijing Foreign Studies University in 1956 and immediately joined the Institute of Geography(now Institute of Geographic Sciences and Natural Resources Research),Chinese Academy of Sciences,as a Russian translator.In 1961,Prof.

The amino acids differences in epitopes may promote the different allergenicity of ovomucoid derived from hen eggs and quail eggs

Quail egg ovomucoid can inhibit activation of basophils and eosinophils,while hen egg ovomucoid has been shown to be a major allergen,named Gal d 1.At present,the differences in structure and function between two ovomucoid are unclear.We found the homology of ovomucoid in quail eggs and hen eggs reached77%.Compared with hen egg ovomucoid,the distribution of secondary structure was different in AA52-53,AA57-58,AA66-68,AA71-72,AA131-133,AA139-140,AA157-159 and AA184-185.Among 9 epitopes of egg ovomucoid,there were different amino acids from quail egg ovomucoid in 8 epitopes.Recombination quail egg ovomucoid had trypsin inhibition activity and quail egg ovomucoid didn't specifically bind to serum of eggs allergic patients.Quail egg ovomucoid can significantly inhibit RBL-2 H3 cells degranulation and protect cells morphology to a certain extent,indicating quail egg ovomucoid can inhibit cells activation and have potential anti-allergic effects,which is related to trypsin inhibitory activity.The difference in sensitization compare to hen egg ovomucoid may be due to amino acids differences affecting protein structure by changing antigenic epitopes.

Effects of Desmodium uncinatum Leaf Meal in the Diet on Lohmann Brown Hens’ Laying Performance and Eggs Quality

A cheap, safe and available alternative protein source to soybean is a huge priority for the developing world. This study is aimed at evaluating the effects of Desmodium uncinatum leaf meal incorporation levels in the diet on Lohmann Brown hens’ laying performances. For this purpose, 60 Lohmann Brown hens aged 33 weeks with an average weight of 1.78 kg were randomly assigned to 3 treatment groups of 20 birds each with 4 replicates. The experimental treatments consisted of DULM 0, DULM5, and DULM10 containing 0, 5 and 10 kg of D. uncinatum leaf meal (DULM) per 100 kg of feed respectively. The results reveal that except for the weights of hens, laying performances were significantly affected (p < 0.05) by DULM in the diet. Daily feed intake was lower with 10% incorporation of DULM in the diet as compared to a diet with 5% DULM which recorded the highest value for this parameter. Moreover, the highest value for the egg weight, egg mass, yolk index, albumen index and egg production were recorded with diet DULM5. On the other hand, the diet with 10% of DULM (DULM10) recorded the highest relative shell weight, feed conversion ratio and egg price. Nevertheless, the mortality rate found in this study was 0% for all treatments confirming the good quality of DULM as protein feedstuff for layer. It was concluded that incorporation levels of D. uncinatum leaves at 5% in the diet improved the laying performances of Lohmann Brown hens.

二乙基亚硝胺诱发C3H/HeN小鼠肝癌模型的研究

[目的]建立C3H/HeN小鼠诱发性肝癌动物模型,为研究肝癌发病机理及开展肝癌药物治疗提供小鼠诱发性肝癌模型。[方法]取20只C3H/HeN小鼠作为模型组,每日喂予含30μg.mL-1二乙基亚硝胺(DEN)的饮水,连续23周后,观察其肝癌的发生情况。另取6只C3H/HeN小鼠作为正常组。[结果]造模23周后,模型组小鼠血清中ALP、ALT、AST、TBIL、肝脏指数、结节数和γ-GT活力显著增加(P<0.01),小鼠体重、血清中TP、ALB、BUN、CREA和肝脏中TP含量显著降低(P<0.01),存活15只小鼠中有13只肉眼能观测到肝癌发生,肝癌发生率为86.67%,小鼠肝脏表面较多结节,体积增大,表面粗糙、色彩暗淡,多个大小不一的灰白色圆形结节,组织学类型为肝细胞癌,其中以梁状型和腺样型居多。[结论]应用DEN诱发的小鼠肝癌,与人肝癌的发生过程相似,是一种较为理想的诱发性肝癌动物模型,但具体实验方法需进一步改良和研究。

Molecular Identification of Gallibacterium from Hen and Analysis of Its 16S rRNA Gene

Two strains of Gallibacterium, isolated from one laying hen flock in Zhengzhou City of Henan Province, were identified by the morphological observation, genus -specific PCR, and analysis of 16S rRNA gene, which was used to generate the phylogenetic tree, with the 21 members of the 12 genera belonging to Pasteurellaceae to analyze the homology. Two strains were named Yu-ZZ-HL-I-SLG and Yu-ZZ-HL-II-GZ. The comparative result of the 16S rDNA sequence shows that the 2 isolated strains are identical in sequence; the highest identity （99.9%） was observed between the isolated strain and one of the strains of Gallibacterium anatis （AF22.8002）, the homologies between the isolated strain and 3 strains of gallibacterium accessed in NCBI （AF22.8016, Gallibacterium genomosp. 1, AF228017, Gallibacterium genomosp. 2, AF22.8018, Gal- libacterium genomosp. 1 ） were above 97.1%, higher than that of the isolated strain and the other strains of the other 11 genera which were between 90.7% -93.2%. It can be seen from the phylogenetic tree that the 2 isolated strains and the other 4 strain of gallibacterium fell into the same branch, furthermore the 2 isolated strains and the strain of Gallibacterium anatis locate in an internal branch, indicating that the 2 isolated strains belong to Gallibacterium anatis.

TLR3和IRF3在华支睾吸虫感染C3H／HeN小鼠中表达的初步分析

目的 建立华支睾吸虫感染小鼠模型,初步分析TLR3、IRF3基因在小鼠肝脏中的表达情况.方法 随机选用健康雌性C3H/HeN小鼠感染华支睾吸虫建立感染模型,分别在感染第0、1、7、14、28、56、84天颈椎脱臼处死小鼠,取肝脏组织进行病理学观察,收集粪便检查华支睾吸虫虫卵.同时,采用逆转录聚合酶链反应（RT-PCR）检测肝脏中Toll样受体（TLRs）中的TLR3和IRF3的mRNA的表达情况.结果 自感染后第20~30天检查出华支睾吸虫虫卵;肝脏病理学结果显示,与正常组小鼠相比,随着时间的增加,感染组小鼠肝脏汇管区由少量炎症细胞浸润、胆管上皮反应性增生发展到在该区域出现大量纤维组织增生伴有炎症细胞浸润,表明华支睾吸虫感染C3H/HeN小鼠模型成功建立.与正常组相比,感染组自感染后第1天起TLR3和IRF3的mRNA表达均升高（P〈0.01）,均在第84天开始下降.结论 华支睾吸虫感染C3H/HeN小鼠后可激活TLR3-TRIF途径,提示TLR3-TRIF途径可能在华支睾吸虫致病过程中起了一定的作用.

利用组织芯片技术研究C57BL/6J和C3H/HeN小鼠放射性肺损伤进程差异及其机制

目的建立C57BL/6J和C3H/HeN两种小鼠放射性肺损伤进程相关组织芯片,并应用其研究FN、LN和a-SMA的表达变化及意义。方法采用20Gy60Coγ射线照射C57BL/6J和C3H/HeN两种小鼠复制动物模型,测定肺组织中羟脯氨酸含量,制备组织芯片,应用免疫组织化学(im-munohistochemistry,IHC)染色方法与图像分析技术定量检测纤连蛋白(fibronectin,FN)、层粘连蛋白(laminin,LN)和a-平滑肌肌动蛋白(alpha-smooth muscle actin,a-SMA)在放射性肺损伤进程中的表达变化。结果组织芯片制备成功,其HE和免疫组织化学染色结果与普通切片具有良好一致性。照射后1~6mC57BL/6J小鼠肺组织病变经历炎症期、增生期和纤维化期,胶原沉积增多,照后1~3mFN表达明显高于正常对照组,照后6m逐渐减少至正常,LN表达在照射后呈渐进性增加,a-SMA表达强于C3H/HeN小鼠;照射后1~6mC3H/HeN小鼠肺组织主要表现为间质性炎症改变,FN表达于照射后1~6m与正常对照组相比无明显变化,LN表达于照射后1~3m明显增强,6m逐渐减少。结论γ射线照...

Histopathological Observation of Colibacillosis in Laying Hens

[Objective] This study aimed to investigate the histopathological changes of hens with colibacillosis.[Method] Epidemiological investigation,clinical symptoms observation,pathological autopsy,laboratory diagnosis and paraffin-embedded tissue section were performed to diagnose the laying hens from a farm in Changli County,affected by a disease similar to pericarditis and perihepatitis in clinical symptoms.[Result] The pathogen isolated from those hens was E.coli.The histopathological changes of the diseased hens included myocardial cell degeneration and inflammatory cell infiltration in heart,fibrotic perihepatitis,necrotic hepatitis in liver,decreased lymphoid cells and inflammatory cell infiltration in spleen,lung congestion,lung room full of erythrocytes,hyaline degeneration in glomerular capillary and small intestinal epithelial cell shedding.[Conclusion] This study will provide scientific basis for the diagnosis and control of colibacillosis in poultry.

Effects of Traditional Chinese Herbal Medicines on Blood Ions and Enzymes of Egg-laying Hens Under Heat Stress

[Objective] This study was conducted to investigate the effects of tradi- tional Chinese herbal medicines on blood biochemical indices of egg-laying hens under heat stress and to provide theoretical basis for clinical application of the medicines. [Method] One hundred of 88-day-old ISA-Brown hens were randomly di- vided into five groups. The hens in four groups were administrated with Prescription I herbal medicines, Prescription II herbal medicines, Vc and blank control under ar- tificially simulated heat stress, respectively. The last group was raised under normal temperature condition. Blood samples of the hens were collected on 1, 4, 8 and 10 d later and detected for their biochemical indices. [Result] The blood sodium （Na）, phosphorus （P）, potassium （K） and chlorine （CI） concentrations in hens under heat stress were decreased, while the activities of alanine aminotransferase （ALT）, lactate dehydrogenase （LDH） and creatine kinase （CK） were increased. These indices grad- ually returned to normal levels after the use of traditional Chinese herbal medicines. [Conclusion] The herbal medicines can relieve the adverse effects caused by heat stress to blood biochemical indices via different metabolic pathways of ions and en- zymes.