凝结芽孢杆菌对感染鸡白痢沙门菌蛋雏鸡生长性能、抗氧化能力和免疫功能的影响

doi:10.16431/j.cnki.1671-7236.2024.10.016

摘要/Abstract

摘要： 【目的】试验旨在研究饲料中添加禽源性凝结芽孢杆菌BC-362对鸡白痢沙门菌(SP)感染蛋雏鸡生长性能、抗氧化能力和免疫功能的影响。【方法】选取150只1日龄健康、体重相近的京红1号SPF蛋雏鸡，随机分为对照组(A组)、益生菌组(B组)、SP+益生菌组(C组)、SP+抗生素组(D组)、SP模型组(E组)，每组6个重复，每个重复5只雏鸡。A、D和E组蛋雏鸡饲喂基础饲粮，B和C组蛋雏鸡在基础饲粮中添加1.0×10⁸ CFU/g凝结芽孢杆菌，7日龄时，C、D和E组蛋雏鸡灌胃1.0×10⁹ CFU SP(0.5 mL)，A和B组蛋雏鸡灌胃同等剂量无菌水，D组蛋雏鸡灌胃后第2天开始，在基础饲粮中添加0.375 g/kg 硫酸新霉素。试验期共14 d。14日龄时测定蛋雏鸡生长性能；采集血液、免疫器官和空肠组织，测定血清抗氧化和免疫指标、免疫器官指数、空肠组织形态、CD3免疫组化表达、炎性和抗氧化相关基因转录水平。【结果】 ①与A组相比，B组蛋雏鸡平均日增重(ADG)显著增加、料重比(F/G)显著下降(P＜0.05)，E组蛋雏鸡平均日采食量(ADFI)和ADG均显著降低(P＜0.05)；与E组相比，C和D组蛋雏鸡ADFI和ADG均显著增加(P＜0.05)；C与D组蛋雏鸡ADFI、ADG和F/R均无显著差异(P＞0.05)。②与A组相比，B组蛋雏鸡血清超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性显著提高(P＜0.05)，E组蛋雏鸡血清SOD、CAT、谷胱甘肽过氧化氢酶(GSH-Px)活性和总抗氧化能力(T-AOC)均显著下降(P＜0.05)，而丙二醛(MDA)含量显著上升(P＜0.05)；与E组相比，C和D组蛋雏鸡血清SOD活性和T-AOC均显著上升(P＜0.05)，MDA含量显著下降(P＜0.05)；C组蛋雏鸡血清中CAT和GSH-Px活性显著高于D组(P＜0.05)。③与A组相比，B组蛋雏鸡血清IgA和IgM含量均显著提高(P＜0.05)，E组蛋雏鸡血清IgA、IgM和IgG含量均显著降低(P＜0.05)；与E组相比，C和D组蛋雏鸡血清IgA、IgM和IgG含量均显著增加(P＜0.05)；C组蛋雏鸡血清IgM含量显著高于D组(P＜0.05)。④与A组相比，B组蛋雏鸡胸腺指数显著提高(P＜0.05)，E组蛋雏鸡免疫器官指数均显著降低(P＜0.05)；与E组相比，C和D组蛋雏鸡胸腺和法氏囊指数显著提高(P＜0.05)；C组蛋雏鸡脾脏指数显著高于D组(P＜0.05)。⑤与A组相比，B组蛋雏鸡空肠组织杯状细胞数量和CD3表达量无显著差异(P＞0.05)，E组蛋雏鸡空肠组织杯状细胞数量和CD3表达量均显著减少(P＜0.05)；与E组相比，C和D组蛋雏鸡空肠组织CD3表达量显著增加(P＜0.05)；C组蛋雏鸡空肠组织杯状细胞数量显著高于D组(P＜0.05)。⑥与A组相比，B组蛋雏鸡空肠组织炎性相关基因Toll样受体4(TLR4)、髓分化因子88(MYD88)、核因子κB(NF-κB)和白细胞介素-6(IL-6)相对表达量显著下调(P＜0.05)，抗氧化相关基因Kelch样ECH关联蛋白1(Keap1)、谷胱甘肽过氧化酶1(GPX1)和谷氨酸半胱氨酸连接酶催化亚基(GCLC)相对表达量显著上调(P＜0.05)，E组蛋雏鸡空肠组织炎性相关基因TLR4、MYD88、IL-1β、NF-κB及IL-6相对表达量均显著上调(P＜0.05)，抗氧化相关基因NADH醌脱氢酶1(NQO1)、血红素氧合酶1(HO-1)、GCLC和核因子E2相关因子(Nrf2)相对表达量均显著下调(P＜0.05)；C和D组蛋雏鸡炎性相关基因TLR4、MYD88、IL-1β、NF-κB和IL-6相对表达量均显著低于E组(P＜0.05)，抗氧化相关基因NQO1和GCLC相对表达量显著高于E组(P＜0.05)；C组蛋雏鸡炎性相关基因TLR4、MYD88和IL-1β相对表达量显著低于D组(P＜0.05)，抗氧化相关基因Keap1和GCLC相对表达量显著高于D组(P＜0.05)。【结论】饲粮中添加禽源性凝结芽孢杆菌BC-362可提高感染SP蛋雏鸡生长性能，增强机体抗氧化及免疫能力，从而减轻SP感染导致的蛋雏鸡肠道炎症反应。

关键词: 凝结芽孢杆菌BC-362; 鸡白痢沙门菌; 生长性能; 抗氧化能力; 免疫功能; 蛋雏鸡

Abstract: 【Objective】 The experiment was conducted to investigate the effects of feed supplementation with avian autochthonous Bacillus coagulans BC-362 on the growth performance，antioxidant capacity and immune function of laying chicks infected with Salmonella Pullorum (SP).【Method】 150 one-day-old healthy Jinghong-1 SPF laying chicks with similar body weights were randomly divided into control group (group A)，probiotic group (group B)，SP + probiotic group (group C)，SP + antibiotic group (group D)，and SP model group (group E). Each group was comprised of 6 replicates，with five chicks per replicate. The basal diet was given to groups A，D，and E，while groups B and C were supplemented with 1.0×10⁸ CFU/g of Bacillus coagulans in the basal diet. At 7 days，groups C，D，and E were gavaged with 1.0×10⁹ CFU of SP (0.5 mL)，and groups A and B were gavaged with the same dose of sterile water. Additionally，group D was supplemented with 0.375 g/kg of neomycin sulphate in the basal diet starting from the second day of gavage. The experimental period lasted for 14 days. At 14 days，the growth performance of the chicks was measured; Blood，immune organs，and jejunum tissues were collected to determine levels of serum antioxidant and immunity，immune organ index，jejunum morphology，CD3 immunohistochemistry expression，and transcript levels of inflammatory and antioxidant-related genes.【Result】 ①Compared with group A，the average daily gain (ADG) of laying chicks in group B was significantly increased，and the feed to gain ratio (F/G) was significantly decreased (P＜0.05). In group E，both the average daily feed intake (ADFI) and ADG of laying chicks were significantly decreased (P＜0.05). Compared with group E，the ADFI and ADG of laying chicks in groups C and D were significantly increased (P＜0.05). There were no significant differences in ADFI，ADG，and F/G of laying chicks between groups C and D (P＞0.05).②Compared with group A，the activities of serum superoxide dismutase (SOD) and catalase (CAT) of laying chicks were significantly higher (P＜0.05) in group B, and the activities of serum SOD，CAT and glutathione peroxidase (GSH-Px) and total antioxidant capacity (T-AOC) of laying chicks were significantly decreased (P＜0.05)，while the malondialdehyde (MDA) content were significantly increased (P＜0.05) in group E.Compared with group E，serum SOD activity and T-AOC of laying chicks were increased significantly (P＜0.05)，and the MDA content of laying chicks were decreased significantly (P＜0.05) in both groups C and D. The activities of serum CAT and GSH-Px of laying chicks in group C were significantly higher than those in group D (P＜0.05). ③Compared with group A，the serum IgA and IgM contents of laying chicks were significantly higher in group B (P＜0.05)，while the serum IgA，IgM and IgG contents of laying chicks were significantly lower in group E (P＜0.05). Compared with group E，the serum IgA，IgM，and IgG contents of laying chicks were significantly increased in groups C and D (P＜0.05)，with the IgM content of laying chicks in group C was significantly higher than that in group D (P＜0.05).④Compared with group A，the thymus index of laying chicks were significantly higher in group B (P＜0.05)，and the immune organ index of laying chicks were significantly lower in group E (P＜0.05). The thymus and bursa indices of laying chicks were significantly higher in groups C and D compared to group E (P＜0.05). The spleen index of laying chicks were significantly higher in group C than that in group D (P＜0.05). ⑤Compared with group A，there was no significant difference in the number of goblet cells and CD3 expression in jejunal tissues of laying chicks in group B (P＞0.05)，while the numbers of goblet cells and CD3 expression were significantly reduced in group E (P＜0.05). The expression of CD3 in jejunal tissues of laying chicks were significantly higher in groups C and D compared to group E (P＜0.05)，and the number of goblet cells in jejunal tissues of laying chicks in group C was significantly higher than that in group D (P＜0.05).⑥Compared with group A，the relative expression of inflammation-related genes Toll-like receptor 4 (TLR4)，myeloid differentiation factor 88 (MYD88)，nuclear factor kappa-B (NF-κB) and interleukin-6 (IL-6) in jejunal tissues of laying chicks were significantly downregulate in group B (P＜0.05)，while the relative expression of antioxidant-related genes Kelch-like ECH associated protein 1 (Keap1)，glutathione peroxidase 1 (GPX1)，and glutamate-cysteine ligase catalytic subunit (GCLC) were significantly upregulate (P＜0.05). The relative expression of inflammation-related genes TLR4，MYD88，IL-1β，NF-κB and IL-6 in jejunal tissues of laying chicks in group E was significantly upregulated (P＜0.05)，while the relative expression of antioxidant-related genes NADH quinone dehydrogenase 1 (NQO1)，heme oxygenase 1 (HO-1)，GCLC，and nuclear factor E2-associated factor (Nrf2) were significantly downregulated (P＜0.05). In groups C and D，the relative expression of inflammation-related genes TLR4，MYD88，IL-1β，NF-κB and IL-6 of laying chicks were significantly lower than those in group E (P＜0.05)，whereas the relative expression of antioxidant-related gene NQO1 and GCLC was significantly higher than in group E (P＜0.05). Compared with group D，the relative expression of TLR4，MYD88，and IL-1β was significantly downregulate (P＜0.05)，and the relative expression of Keap1 and GCLC was significantly upregulate in group C (P＜0.05).【Conclusion】 The supplementation of avian autochthonous Bacillus coagulans BC-362 to diets improved the growth performance of SP-infected laying chicks and enhanced the body’s antioxidant and immune capabilities，thereby reducing the intestinal inflammatory response infected with SP in laying chicks.

Key words: Bacillus coagulans BC-362; Salmonella Pullorum; growth performance; antioxidant capacity; immune function; laying chicks

中图分类号:

S831.5

田光明, 马丽, 蒲跃进, 游蕾, 王皓杰, 张应胡, 方春, 梁雄燕, 温振宇, 谭磊, 郑明浩. 凝结芽孢杆菌对感染鸡白痢沙门菌蛋雏鸡生长性能、抗氧化能力和免疫功能的影响[J]. 中国畜牧兽医, 2024, 51(10): 4352-4363.

TIAN Guangming, MA Li, PU Yuejin, YOU Lei, WANG Haojie, ZHANG Yinghu, FANG Chun, LIANG Xiongyan, WEN Zhenyu, TAN Lei, ZHENG Minghao. Effect of Bacillus coagulans on Growth Performance，Antioxidant Capacity and Immune Function of Laying Chicks Infected with Salmonella Pullorum[J]. China Animal Husbandry and Veterinary Medicine, 2024, 51(10): 4352-4363.

参考文献

[1] 陆瑶,丁仕豪,曹俊,等.鸡白痢沙门氏菌PCR检测方法的建立[J].中国预防兽医学报,2021,43(11):1178-1183.LU Y，DING S H，CAO J，et al. Establishment of a specific PCR method for detection of Salmonella Pullorum[J]. Chinese Journal of Preventive Veterinary Medicine，2021,43(11): 1178-1183. (in Chinese)
[2] BARROW P A，NETO O C. Pullorum disease and fowl typhoid—New thoughts on old diseases: A review[J]. Avian Pathology，2011,40(1):1-13.
[3] KOCOT A M，JAROCKA-CYRTA E，DRABINSKA N，et al. Overview of the importance of biotics in gut barrier integrity[J]. International Journal of Molecular Sciences，2022,23(5):2896.
[4] YANG L，CHEN Y，BAI Q，et al. Protective effect of bifidobacterium lactis JYBR-190 on intestinal mucosal damage in chicks infected with Salmonella Pullorum[J]. Frontiers in Veterinary Science，2022,9:879805.
[5] WANG Y，LI J，XIE Y，et al. Effects of a probiotic-fermented herbal blend on the growth performance，intestinal flora and immune function of chicks infected with Salmonella Pullorum[J]. Poultry Science，2021,100(7):101196.
[6] DAS S，SEN R. Kinetic modeling of sporulation and product formation in stationary phase by Bacillus coagulans RK-02 vis-à-vis other Bacilli[J]. Bioresource Technology，2011,102(20):9659-9667.
[7] CAO J，YU Z，LIU W，et al. Probiotic characteristics of Bacillus coagulans and associated implications for human health and diseases[J]. Journal of Functional Foods,2020,64:103643.
[8] WANG X，JIAN H，ZHAO W，et al. Effects of dietary Bacillus coagulans on the productive performance，egg quality，serum parameters，and intestinal morphology of laying hens during the late laying period[J]. Italian Journal of Animal Science，2023,22(1):95-105.
[9] ZHANG Y，TIAN X，DONG Y，et al. Bacillus coagulans prevents the decline in average daily feed intake in young piglets infected with enterotoxigenic Escherichia coli K88 by reducing intestinal injury and regulating the gut microbiota[J]. Frontiers in Cellular and Infection Microbiology，2023,13:1284166.
[10] MARIANA S，VALFREDO S，FILHO T，et al. The step of incorporation of Bacillus coagulans GBI-306086 into "requeijão cremoso" processed cheese does not affect metabolic homeostasis of rats[J]. Frontiers in Microbiology，2019,10:2332.
[11] ZHENG W，ZHAO Z，YANG Y，et al. The synbiotic mixture of lactulose and Bacillus coagulans protects intestinal barrier dysfunction and apoptosis in weaned piglets challenged with lipopolysaccharide[J]. Journal of Animal Science and Biotechnology，2023,14(1):80.
[12] STECKER R A，MOON J M，RUSSO T J，et al. Bacillus coagulans GBI-30，6086 improves amino acid absorption from milk protein[J]. Nutrition & Metabolism，2020,17(1):93.
[13] HU R G，YANG B T，ZHENG Z Y，et al. Improvement of non-specific immunity，intestinal health and microbiota of crucian carp (Carassius auratus) juvenile with dietary supplementation of Bacillus coagulans BC1[J]. Aquaculture，2024,580(2):740327.
[14] DENG Z，HAN D，WANG Y，et al. Lactobacillus casei protects intestinal mucosa from damage in chicks caused by Salmonella Pullorum via regulating immunity and the Wnt signaling pathway and maintaining the abundance of gut microbiota[J]. Poultry Science，2021,100(8):101283.
[15] VITETTA L，BRISKEY D，ALFORD H，et al. Probiotics，prebiotics and the gastrointestinal tract in health and disease[J]. Inflammopharmacology，2014,22(3):135-154.
[16] SANDERS M E，MORELLI L，TOMPKINS T A. Sporeformers as human probiotics: Bacillus，Sporolactobacillus，and Brevibacillus[J]. Comprehensive Reviews in Food Science and Food Safety，2006,2(3):101-110.
[17] 肖明霞.饲用凝结芽孢杆菌对热应激肉鸡肠道营养消化吸收的影响及作用机制[D].昆明：云南农业大学,2023.XIAO M X. Effects and mechanism of feeding Bacillus coagulans on intestinal function of digestion and absorption of broiler chickens in heat stress[D]. Kunming:Yunnan Agricultural University，2023. (in Chinese)
[18] HOU J，LIAN L，LU L，et al. Effects of dietary Bacillus coagulans and tributyrin on growth performance，serum antioxidants，intestinal morphology，and cecal microbiota of growing Yellow-feathered broilers[J]. Animals，2023,13(22):3534.
[19] 江科,李和刚,戴正浩,等.益生菌在动物养殖业中应用的研究进展[J].中国畜牧兽医，2013，40(12)：90-94.JIANG K,LI H G,DAI Z H，et al. Research progress on the application of probiotics in animal breeding[J]. China Animal Husbandry & Veterinary Medicine，2013，40(12)：90-94. (in Chinese)
[20] SCHIEBER M，CHANDEL N S. ROS function in redox signaling and oxidative stress[J]. Current Biology，2014,24(10):453-462.
[21] LU T，WANG L，WU Q，et al. Bacillus coagulant HYI (BC-HYI) alleviates LPS-elicited oxidative stress by engaging the Nrf2/HO-1 signaling pathway and regulates gut macrobiotics in laying chickens[J]. Applied Microbiology，2023,3(4):1178-1194.
[22] THIRUNAVUKKARASU M，PENUMATHSA S V，KONERU S，et al. Resveratrol alleviates cardiac dysfunction in streptozotocin-induced diabetes: Role of nitric oxide，thioredoxin，and heme oxygenase[J]. Free Radical Biology and Medicine，2007,43(5):720-729.
[23] YANG G，SHEN K，YU R，et al. Probiotic (Bacillus cereus) enhanced growth of Pengze crucian carp concurrent with modulating the antioxidant defense response and exerting beneficial impacts on inflammatory response via Nrf2 activation[J]. Aquaculture，2020,529:735691.
[24] BIAN X，WALLSTROM G，DAVIS A，et al. Immunoproteomic profiling of antiviral antibodies in new-onset type 1 diabetes using protein arrays[J]. Diabetes，2016,65(1):285-296.
[25] 朱随亮,李玥,郑旭,等.抗菌肽、酸化剂和功能性寡糖联合添加对肉仔鸡生长性能、肠道形态以及盲肠微生物区系的影响[J].动物营养学报，2021,33(8):4424-4437.ZHU S L,LI Y,ZHENG X，et al. Effects of antimicrobial peptides，acidifiers and functional oligosaccharide combined supplementation on growth performance，intestinal morphology and cecal microflora of broilers[J]. Chinese Journal of Animal Nutrition，2021,33(8): 4424-4437. (in Chinese)
[26] ZHAO Z，SUN M，CUI X，et al. Bacillus coagulans Mzy531 alleviates intestinal mucosal injury in immunosuppressive mice via modulating intestinal barrier，inflammatory response，and gut microbiota[J]. Scientific Reports, 2023,13(1):11181.
[27] GUO M，HAO G，WANG B，et al. Dietary administration of Bacillus subtilis enhances growth performance，immune response and disease resistance in Cherry Valley ducks[J]. Frontiers in Microbiology，2016,7:1975.
[28] GEORGE M,BIRCHENOUGH E E，MALIN E，et al. A sentinel goblet cell guards the colonic crypt by triggering Nlrp6-dependent Muc2 secretion[J]. Science，2016,352(6293):1535-1542.
[29] FORTE C，MANUALI E，ABBATE Y，et al. Dietary Lactobacillus acidophilus positively influences growth performance，gut morphology，and gut microbiology in rurally reared chickens[J]. Poultry Science，2018,97(3):930-936.
[30] AMER S A，GOUDA A，SALEH G K，et al. Dietary frankincense (Boswellia serrata) oil modulates the growth，intestinal morphology，the fatty acid composition of breast muscle，immune status，and immunoexpression of CD3 and CD20 in broiler chickens[J]. Animals，2023,13(6):971.
[31] ZHOU J，ZHANG W，LIU W，et al. Histological study of intestinal goblet cells，IgA，and CD3⁺ lymphocyte distribution in Huang-huai White goat[J]. Folia Morphologica，2020,79(2):303-310.
[32] QIAO J，SUN Z，LIANG D，et al. Lactobacillus salivarius alleviates inflammation via NF-κB signaling in ETEC K88-induced IPEC-J2 cells[J]. Journal of Animal Science and Biotechnology，2020,11(1):76.
[33] LIN Y L，YANG X G，YUE W J，et al. Chemerin aggravates DSS-induced colitis by suppressing M2 macrophage polarization[J]. Cellular & Molecular Immunology，2014,11(4): 355-366.
[34] 李静,孙洋,熊心雨,等.樱黄素抑制TLR4/MyD88通路减轻肠上皮炎症反应改善小鼠克罗恩病样结肠炎[J].细胞与分子免疫学杂志,2024,40(3):199-206.LI J，SUN Y，XIONG X Y，et al. Prunetin inhibits TLR4/MyD88 pathway to attenuate intestinal epithelial inflammatory response and ameliorate mouse Crohn’s disease-like colitis[J].Chinese Journal of Cellular and Molecular Immunology，2024,40(3):199-206. (in Chinese)
[35] LIU R，CHEN H Q，REN W J，et al. Ginseng (Panax ginseng Meyer) oligopeptides protect against binge drinking-induced liver damage through inhibiting oxidative stress and inflammation in rats[J]. Nutrients,2018,10(11):1665.