摘要
为研究发酵鸡球虫散对海兰褐蛋鸡感染柔嫩艾美耳球虫的治疗作用,本实验选取90只健康雏鸡,随机分为6组,分别为I对照组、II感染组、III益生菌组(植物乳杆菌菌液)、IV中药组(0.2%鸡球虫散)、V发酵组(0.2%发酵鸡球虫散)和VI西药组(0.02%地克珠利),于8日龄时III~VI组鸡分别饲喂基础日粮和相应药剂,16日龄时,除I对照组外,其余各组雏鸡均灌胃感染球虫,其中发酵鸡球虫散为经植物乳杆菌发酵的经典方剂鸡球虫散。感染后每天观察各组雏鸡临床症状,记录并计算粪便中的球虫卵囊值。感染后9 d剖杀各组鸡,剖杀前8 h禁食并称重,采集血清、免疫器官和盲肠组织,计算各组鸡相对增重率、存活率、计算球虫卵囊值和抗球虫指数(ACI),以及器官指数。观察各组雏鸡盲肠组织形态学变化,利用ELISA试剂盒检测各组鸡血清中各类抗体、炎性因子及抗氧化指标,采用网络药理学分析中药复方成分治疗疾病的潜在关键靶点等,采用荧光定量PCR检测各组鸡盲肠组织中TLR4、MyD88、NF-κB、p65、Occludin以及ZO-1基因相对转录水平的变化。结果显示,与I对照组相比,II感染组鸡表现出明显的临床症状,精神委顿、嗜睡等,粪便检查可见鸡球虫卵囊,并伴随不同程度血便,剖检死亡鸡可见盲肠红肿;相对增重率、存活率、ACI均低于I对照组;器官指数(脾脏、胸腺、法氏囊)均极显著降低(P<0.01);盲肠组织绒毛不完整、断裂,绒毛高度/隐窝深度比值极显著性降低(P<0.01);血清中IgA、IgM、IgG含量和IL-6、IFN-γ、TNF-α含量均极显著升高(P<0.01),而CAT与SOD活性、GSH-Px与IL-10含量均极显著降低(P<0.01),MDA含量极显著升高(P<0.01);与II感染组相比,以上指标在III益生菌组、IV中药组、V发酵组和VI西药组均得到不同程度缓解,其中V发酵组的缓解作用最为明显。经网络药理学分析发现,疾病与药物交集靶点38个,构建鸡球虫散-成分-球虫病-靶点网络图;蛋白互作网络图(PPI)分析到34个存在相互作用的蛋白质核心靶点;经GO与KEGG分析显示蛋白互作涉及20条相关通路,主要富集于免疫相关生物过程和炎症性肠病等通路。荧光定量PCR检测结果显示,与I对照组相比,II感染组盲肠组织中TLR4、MyD88、NF-κB和p65的相对转录水平均极显著升高(P<0.01),Occludin和ZO-1的相对转录水平极显著降低(P<0.01)。其他组上述指标均得到改善,尤其发酵组的改善最明显。上述结果首次证实植物乳杆菌发酵可显著提高鸡球虫散的综合药效,发酵鸡球虫散可能通过干预TLR4/MyD88/NF-κB信号传导途径,调控宿主炎症介质表达,提高蛋鸡抗球虫指数,缓解柔嫩艾美耳球虫感染导致的蛋鸡免疫器官损伤及氧化应激反应,同时促进肠黏膜的再生与修复。本研究为发酵鸡球虫散的临床应用提供了科学依据。
To study the therapeutic effect of fermented chicken coccidiostat on the infection of soft Eimeria coccidia in Hyland Brown laying hens,in this experiment,90 healthy chicks were selected and randomly divided into 6 groups,which were I control group,II infected group,III probiotic group(Lactobacillus plantarum bacterial suspension),IV traditional Chinese medicine group(0.2%chicken Coccidiosis San),V fermentation group(0.2%fermented chicken Coccidiosis San)and VI western medicine group(0.02%diclazuril).On 8 days of age,groups III,IV,V and VI were fed basal diet+4%probiotics,basal diet+0.2%coccidiosis powder,basal diet+0.2%fermented chicken coccidioides,and basal diet+0.02%diclazuril,respectively.On 16 days of age,except for the I control group,all other groups were infected with coccidioides by gavage.The number of coccidia oocysts in faeces was recorded daily after infection and clinical symptoms were observed.The animals were dissected at 9 days post-infection(24 days old)and were fasted and weighed 8 hours before execution.Serum,immune organs and cecum tissues were collected to calculate the relative weight gain rate and survival rate of chickens in each group and the oocyst value and anti-coccidial index(ACI).Morpho-logical changes in cecum tissues were observed,and ELISA kits were used to detect various types of antibodies,inflammatory factors and antioxidant indicators in serum,network pharmacology was used to analyse the potential key targets of the TCM compound ingredients for the treatment of diseases,and fluorescence quantitative PCR was used to determine the changes in the relative transcript levels of the cecropin genes TLR4,MyD88,NF-κB,p65,Occludin,and ZO-1.The results showed that compared with the I control group,chickens in the II infected group showed obvious clinical symptoms,such as mental agitation and lethargy;fecal examination showed fowl coccidia oocysts accompanied by varying degrees of bloody stools,and cecum redness and swelling were seen in dead chickens by autopsy;the relative weight gain rate,survival rate,and ACI were lower than those in the I control group;organ indices(spleen,thymus,and bursa of fasciola gigantica)were highly significantly lower(P<0.01);incomplete and broken villi appeared,and the ratio of villus height/crypt depth was highly significantly lower(P<0.01);serum antibody levels of IgA,IgM,IgG and levels of IL-6,IFN-γand TNF-αwere highly significantly higher(P<0.01),whereas the levels of CAT and SOD activity,GSH-Px and IL-10 were highly significantly lower(P<0.01),and the level of MDA was highly significantly higher(P<0.01);the relative transcript levels of TLR4,MyD88,NF-κB and p65 in cecum tissues were highly significantly increased(P<0.01),and the relative transcript levels of Occludin and ZO-1 were highly significantly decreased(P<0.01).Compared with the II infection group,the above indicators were alleviated to different degrees in the III probiotic group,IV traditional Chinese medicine group,V fermentation group and VI western medicine group,with the alleviating effect of the V fermentation group being particularly prominent.After network pharmacological analysis,38 targets at the intersection of disease and drug were identified,and a network map of chicken Coccidia San-components-coccidiosis-targets was constructed;34 interacting protein core targets were identified by PPI analysis;and 20 relevant pathways were obtained by GO and KEGG analyses,which were mainly enriched in the immune-related bioprocesses and inflammatory bowel disease and other pathways.These results provide the first evidence that Lactobacillus plantarum fermentation can significantly enhance the comprehensive efficacy of chicken coccidiostat.Fermented chicken coccidiostat may alleviate the damage to immune organs and oxidative stress response in laying hens caused by Eimeria tenella infection,and promote intestinal mucosa regeneration and repair,potentially by intervening in the TLR4/MyD88/NF-κB signaling pathway and regulating the expression of inflammatory mediators,thereby improving the anticoccidial index.This study provides a scientific basis for the clinical application of fermented chicken coccidiostat.
作者
王旭贞
李豪
张静頔
郝松华
王淑芳
随志文
范新凤
孙子龙
WANG Xu-zhen;LI Hao;ZHANG Jing-di;HAO Song-hua;WANG Shu-fang;SUI Zhi-wen;FAN Xin-feng;SUN Zi-long(Shanxi Animal Husbandry and Veterinary School,Taiyuan 030002,China;College of Veterinary Medicine,Shanxi Agricultural University,Jinzhong 030801,China;College of Life Sciences,Shanxi Agricultural University,Jinzhong 030801,China)
出处
《中国预防兽医学报》
北大核心
2025年第9期902-912,共11页
Chinese Journal of Preventive Veterinary Medicine
基金
山西省基础研究计划项目(202103021224007)。
关键词
发酵中药
鸡球虫散
柔嫩艾美耳球虫
抗氧化
炎性因子
fermented Chinese medicine
chicken Coccidia San
Eimeria tenella
anti-oxidation
inflammatory factor
