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Effects of Lactobacillus plantarum on production performance,immune characteristics,antioxidant status,and intestinal microflora of bursin-immunized broilers

Effects of Lactobacillus plantarum on production performance...

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中国畜牧兽医学会动物微生态学分会第五届第十二次全国学术研讨会

作者： Xuejiao Shen Dan Yi Xueqin Ni Dong Zeng Bo Jing Mingxia Lei Zhengrong Bian Yan Zeng Tao Li Jinge Xin Key Laboratory of Animal Disease and Human Health of Sichuan Province College of Veterinary Medicine Sichuan Agricultural University Chengdu 611130 China

Examples of probiotic that can promote host health by improving its intestinal microbial balance and intestinal immunity belong to the genus *** (BS) is a peptide isolated from bursa of Fabricius (BF) as an adjuvant for a variety of *** investigate the synergistic effects of Lactobacillus plantarum (LP) dietary supplementation and BS immunization on production performance, immune characteristics, antioxidant status, and intestinal microflora in broilers, 200 one-day-old broilers of mixed sex were randomly allocated into 4 treatments in a 2×2 factorial arrangement (LP-/BS-, LP-/BS+, LP+/BS-, LP+/BS+) for 42 *** immunization enhanced immune response by increasing serum total IgG concentration and IL-6 concentration, promoted antioxidant capacity by increasing CAT activities in serum and liver and decreased serum MDA content at 42 day of age (DOA), enriched intestinal microflora *** supplementation enhanced immune response by increased IL-2 concentration at 42 DOA, promoted antioxidant capacity by increasing liver CAT activities, increasing GSH-Px activities in serum and liver at 21 DOA, and decreasing serum MDA content at 42 DOA, promoted intestinal mieroflora composition by decreasing total aerobes and *** counts at 21 DOA, increasing total anaerobes count at 21 DOA, increasing Lactobacillus *** Bifidobacterium *** at both 21 and 42 *** were significant interactions between BS and LP on daily body weight gain (BWG) and feed conversion ratio (FCR) in the starter period (1 to 21 DOA), IL-2 concentration and liver MDA content at 21 DOA, thymus index (TI), peripheral lymphocyte proliferation and *** counts at 42 ***, these data suggested that the combination of LP dietary supplementation and BS immunization promoted the production performance, immune characteristics, antioxidant status, and intestinal microflora of broilers.

关键词： broiler Lactobacillus plantarum bursin production performance immune characteristic antioxidant status intestinal microflora

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低聚壳聚糖对断奶仔猪抗氧化功能的影响

低聚壳聚糖对断奶仔猪抗氧化功能的影响

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中国畜牧兽医学会动物微生态学分会第五届第十二次全国学术研讨会

作者：王玉郑春田温晓鹿胡胜兰马现永王丽杨雪芬广东省农业科学院动物科学研究所农业部华南动物营养与饲料重点实验室畜禽育种国家重点实验室广东省动物育种与营养公共实验室广东省畜禽育种与营养重点实验室

本试验拟探讨日粮中添加低聚壳聚糖对断奶仔猪抗氧化功能的影响。选取21日龄断奶的杜×(长×大)(体重约6kg)三元杂交去势公猪100头,按照体重将所有试验猪随机分配到5个处理组,每个处理组5个重复,每个重复4头猪。分组如下:(1)CTR... 详细信息

本试验拟探讨日粮中添加低聚壳聚糖对断奶仔猪抗氧化功能的影响。选取21日龄断奶的杜×(长×大)(体重约6kg)三元杂交去势公猪100头,按照体重将所有试验猪随机分配到5个处理组,每个处理组5个重复,每个重复4头猪。分组如下:(1)CTR组:基础饲粮;(2)AZ组;基础饲粮基础上添加30 mg/kg金霉素,12 mg/kg硫酸抗敌素,3000 mg/kg氧化锌;(3)1/2AZ-LC组:基础饲粮基础上添加15 mg/kg金霉素,6 mg/kg硫酸抗敌素,1500 mg/kg氧化锌,50 mg/kg低聚壳聚糖;(4)1/2Z-LC组:基础饲粮基础上添加1500 mg/kg氧化锌,50 mg/kg低聚壳聚糖);(5)LC组:基础饲粮基础上添加50 mg/kg低聚壳聚糖。试验周期28天,试验期间仔猪自由采食和饮水,各试验组所处环境条件及饲养管理均保持一致。第28天饲养试验结束,每个重复选取一头仔猪进行前腔静脉采血,分离血清,测定血清中丙二醛(MDA)和抗氧化酶的活性,。结果显示:(1)与对照组相比,添加低聚壳聚糖的试验组能够显著提高试验猪的平均日增重(ADG);(2)与对照组相比,添加低聚壳聚糖总体上可提高仔猪血清过氧化氢酶(CAT)活性,其中1/2AZ-LC与空白组单独相比,差异达到统计学显著水平(P<0.05)。与空白组相比,添加低聚壳聚糖和/或抗生素总体上可提高仔猪血清总抗氧化酶(TAOC)活性,其中AZ组、1/2AZ-LC组、1/2Z-LC组与空白组单独比较,差异达到统计学显著水平(P<0.05)。与空白组相比,AZ组、1/2AZ-LC组、1/2Z-LC组和LC组仔猪血清丙二醛(MDA)含量分别降低了11%、43%、48%和44%(P>0.05)。仔猪血清总超氧化物歧化酶(TSOD)和谷胱甘肽过氧化物酶(GSH-Px)活性各组间均无显著差异(P>0.05)。研究结果表明:日粮中低聚壳聚糖的添加可提高仔猪血清抗氧化的活性,进而提高机体的抗氧化能力。

关键词：低聚壳聚糖断奶仔猪酶活抗氧化

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Fermented Yupingfeng Polysaccharides Enhance Immunity by Improving the Foregut Microflora and Intestinal Barrier in Weaning Rex Rabbits

Fermented Yupingfeng Polysaccharides Enhance Immunity by Imp...

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中国畜牧兽医学会动物微生态学分会第五届第十二次全国学术研讨会

作者： Hao Sun Xueqin Ni Xu Song Bin Wen Yi Zhou Fuqin Zou Mingyue Yang Zhirong Peng Hui Zhu Yan Zeng Hesong Wang Xiangchao Fu Yunduo Shi Zhongqiong Yin Kangcheng Pan Bo Jing Dong Zeng Ping Wang Animal Microecology Institute College of Veterinary MedicineSichuan Agricultural University Key Laboratory of Animal Disease and Human Health of Sichuan Province Sichuan Academy of Grassland Science Sichuan Fishery School

Yupingfeng(YPF) is a kind of Astragali Radix-based ancient Chinese herbal supplemented with Atractylodis Macrocephalae Rhizoma and Radix *** evidence has proven the beneficial immunomodulating activity of ***,the action mechanism(s) of it is not ***,we explored the immunomodulatory activity of unfermented Yupingfeng polysaccharides(UYP) and fermented Yupingfeng polysaccharides(FYP) obtained using Rhizopus oligosporus SH in weaning rex *** results showed that both UYP and FYP exhibited notable growth-promoting and immune-enhancing activities,improvement of the intestinal flora homeostasis and maintenance of intestinal barrier integrity and ***,compared with UYP,FYP effectively enhanced average daily gain,organ indices,IL-2,IL-4,IL-10,TNF-α,TLR2 and TLR4 mRNA levels in spleen,IL-1,IL-2,IL-4,IL-6,IL-10,IL-12,TNF-a and IFN-γ protein concentrations in serum and TLR2 and TLR4 mRNA expressions in gastrointestinal tract(GIT).Moreover,FYP exhibited greater beneficial effects in improving the intestinal flora,including augment flora diversity and the abundance of cellulolytic bacteria,reduction the abundance of Streptococcus *** Enterococcus *** GIT,particularly foregut and maintaining the intestinal barrier integrity and functionality by upregulating zonula occludens 1,claudin,polymeric immunoglobulin receptor,trefoil factor and epidermal growth factor mRNA levels in jejunum and *** results indicated the immunoenhancement effect of FYP is superior over that of UYP,which is probably related with the amelioration of the intestinal microflora and intestinal barrier in foregut.

关键词： Yupingfeng polysaccharide Rhizopus oligosporus SH Immunity Intestinal microflora Intestinal barrier Toll-like receptors

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Bacillus amyloliquefaciens SC06 alleviates the oxidative stress of IPEC-1 via modulating Nrf2/Keap1 signaling pathway and decreasing ROS production

Bacillus amyloliquefaciens SC06 alleviates the oxidative str...

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中国畜牧兽医学会动物微生态学分会第五届第十二次全国学术研讨会

作者： Yang Wang Yanping Wu Yibing Wang Aikun Fu Li Gong Weifen Li Yali Li Key Laboratory of Molecular Animal Nutrition of the Ministry of Education Institute of Feed ScienceCollege of Animal Sciences Zhejiang University

Oxidative stress（OS） plays a major role in the gastrointestinal *** probiotics were reported to repress OS,few researches compared the antioxidant ability of different Bacillus strains and decipher the *** select a Bacillus strain with higher antioxidant capacity,we used HO to induce intestinal porcine epithelial cell 1（IPEC-1） OS *** most suitable HO concentration and incubation time were determined by the half lethal dose and Methyl Thiazolyl *** analysis was performed to choose a sensitive indicator for *** for the comparison of Bacillus,cells were divided into:control,Bacillus treatment,HO treatment and Bacillus pre-protection+HO *** were co-cultured with IPEC-1 for 3h in Bacillus and Bacillus pre-protection+HO ***,based on OS model,300 μmol/L HO were added into medium of HO and Bacillus pre-protection+HO treatments for another 12 *** and apoptosis gene expressions were detected to screen the target *** factor erythroid-derived 2 related factor 2（Nrf2）/Kelch-like ECH-associated protein 1（Keap1） pathway,reactive oxygen species（ROS） production,mitochondrial membrane potential（△ψm）,apoptosis and necrosis were *** revealed that heme oxygenase-1（HO-1） gene expression had a positive correlation with HO ***,Bacillus amyloliquefaciens SC06（SC06）-meditated IPEC-1 showed the best antioxidant capacity though modulating Nrf2 phosphorylation.△ψm was elevated while ROS generation was reduced with SC06 pre-protection,resulting in decreased apoptosis and ***,HO-1 expression could be regarded as an OS *** regulation of Nrf2/Keap1 pathway and ROS production by SC06 are involved in alleviating OS of IPEC-1.

关键词： Bacillus IPEC-1 Oxidative stress Nrf2 ROS NOX

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Investigation of TbfA in Riemerella anatipestifer using plasmid-based methods for gene over-expression and knockdown

Investigation of TbfA in Riemerella anatipestifer using plas...

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中国畜牧兽医学会动物微生态学分会第五届第十二次全国学术研讨会

作者： MaFeng Liu MengYi Wang DeKang Zhu MingShu Wang RenYong Jia Shun Chen KunFeng Sun Qiao Yang Ying Wu XiaoYue Chen Francis Biville AnChun Cheng Institute of Preventive Veterinary Medicine Sichuan Agricultural University Avian Disease Research Center College of Veterinary Medicine of Sichuan Agricultural University Key Laboratory of Animal Disease and Human Health of Sichuan Province Sichuan Agricultural University Unite des Infections Bacteriennes Invasives Departement Infection et EpidemiologieInstitut Pasteur

Riemerella anatipestifer is a duck pathogen that has caused serious economic losses to the duck industry *** this,there are few reported studies of the physiological and pathogenic mechanisms of Riemerella anatipestifer *** previous study,we have shown that TonB1 and TonB2 were involved in hemin *** family protein（TbfA） was not investigated,since it was failed to knockout at that ***,we used a plasmid based gene overexpression and knockdown to investigate ***,we constructed three Escherichia-Riemerella anatipestifer shuttle vectors containing three different native Riemerella anatipestifer *** shuttle plasmids were introduced into Riemerella anatipestifer ATCC11845 by conjugation at an efficiency of 5×10 antibiotic-resistant transconjugants per recipient *** on the high-expression shuttle vector pLMF03,a method for gene knockdown was *** of TbfA in Riemerella anatipestifer ATCC11845 decreased the organism's growth ability in TSB medium but did not affect its hemin *** contrast,over-expression of TbfA in Riemerella anatipestifer ATCC11845△tonB1△tonB2 significantly promoted the organism's growth in TSB medium but significantly inhibited its hemin ***,these findings suggest that TbfA is not involved in hemin utilization by Riemerella anatipestifer.

关键词： Riemerella anatipestifer TonB family protein(TbfA) Gene overexpression Gene knockdown Investigate

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Effect of green forage on development of the reproductive organ and diversity of the gut microbiota in Meishan prepubertal gilts

Effect of green forage on development of the reproductive or...

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中国畜牧兽医学会动物微生态学分会第五届第十二次全国学术研讨会

作者： Meng Cao Pan Zhang Yang Li Shengyu Xu Yan Lin Jian Li Bin Feng Lianqiang Che Zhengfeng Fang De Wu Key Laboratory for Animal Disease-Resistant Nutrition of the Ministry of Education of China Animal Nutrition InstituteSichuan Agricultural University

This study was conducted to evaluate the effects of green forage on the onset of puberty and estrus cycle,the development of reproductive organ and diversity of the gut microbiota in Meishan prepubertal *** purebred prepubertal Meishan gilts(ages 93.3 ± 6.8 d;initial body weight 25.3 ± 0.8 kg)were randomly allocated to one of two groups with 10 repeats for each group:control(CON) and Green Forage group(GF).The control basal diets were formulated to meet or exceed Chinese Feeding Standard of Swine(2004).In Green Forage group we used Chinese cabbage instead of part of basal diet with almost iso-energetic and *** control group gilts fed 1.08 kg,1.30 kg,1.64 kg of basal diets(corn-soybean meal diets) per day at stage 1(20-3 0kg),stage 2(30-40kg) and stage 3(40kg to the fourth estrus),*** Green Forage group gilts were fed 1.2kg,1.8kg,2.4kg of Chinese cabbage,and1.02 kg,1.21 kg,1.54 kg of basal diets at stage 1(20-30kg),stage 2(30-40kg) and stage 3(40kg to the fourth estrus),*** age at first estrus of gilts fed green forage was delayed compared with the control gilts(152.4 d vs 135.2 d,P = 0.003).The number of large follicles(>5mm) were increased compared with the control gilts(16.4 vs 12.6,P = 0.041).The concentration of IGF-1 in follicular fluid was increased significantly compared with the control *** was no effect of green forage on estrus cycle and duration of *** gilts fed green forage had higher levels of intestinal microbial richness and biodiversity than the control *** study demonstrates that the green forage can promote the development of reproductive organ in Meishan prepubertal ***,this may be a hint that the improvement of the reproductive organs may be associated with the change of intestinal microorganism.

关键词： gilts green forage reproductive organ intestinal microorganism

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PCR-DGGE手段鉴定伪无菌小鼠定植大熊猫肠道菌群的效果研究

PCR-DGGE手段鉴定伪无菌小鼠定植大熊猫肠道菌群的效果研究

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中国畜牧兽医学会动物微生态学分会第五届第十二次全国学术研讨会

作者：赖婧倪学勤曾东王鹤松潘康成景波四川农业大学动物医学院动物微生态研究中心

大熊猫(Ailuropoda melanoleuca)作为一种被高度重视的保护动物,完善对其肠道菌群的研究手段对于大熊猫生理机能、病理分析及药物研发等领域均具有极为重要的作用。然而近年来尽管对大熊猫的肠道菌群研究日益深入,由于其自身的稀有及政... 详细信息

大熊猫(Ailuropoda melanoleuca)作为一种被高度重视的保护动物,完善对其肠道菌群的研究手段对于大熊猫生理机能、病理分析及药物研发等领域均具有极为重要的作用。然而近年来尽管对大熊猫的肠道菌群研究日益深入,由于其自身的稀有及政策的限制,绝大部分研究依旧集中于对大熊猫所排泄出的粪便,并以此推测动物机体内部的情况,这一限制直接导致研究的严谨性受损,并彻底限制了对其菌群结构进行人为调控研究的可能性。近年来,通过转移某种动物的肠道菌群至另外一类更易操作的动物体内,形成诸如肠道菌群人源化小鼠等技术开始逐渐成熟,并提供合理的参考。在珍稀动物领域,该方法为大熊猫等保护动物进行在体实验提供了一种新的方式和可能。本实验使用抗生素处理小鼠的肠道来得到伪无菌小鼠,并接种定植大熊猫的肠道菌群,结合PCR-DGGE(polymerase chain reaction-denatured gradient gel electrophoresis)技术,旨在得到更符合大熊猫肠道菌群特征的小鼠模型,为开展研究研究大熊猫的体内机制探寻动物模型并提供理论依据。本试验选择了10只成年健康大熊猫的粪便样本(来自成都大熊猫繁育研究基地),提取各自的粪便菌群DNA以及混合样本后的粪便菌群DNA,利用PCR-DGGE技术,得到条带后进行聚类分析,分别分析其菌群结构的相似性和多样性之间的差别;选择20只四周龄左右雌性昆明小鼠,饲喂一周缓冲后,连续灌喂五天链霉素,一日两次,随后分为两组(单次组及多次组),分别接种一次及三次(每日一次)混合的大熊猫粪便细菌悬液,选取造模后第三天的粪便,进行PCR-DGGE分析,比对两试验组以及大熊猫粪便菌群的菌群结构相似性及多样性差异。试验结果如下:1.混合后的熊猫粪便菌群均一性高,且相似度(>95%)远高于单只熊猫的平均相似度(74%),更加符合造模的条件,因此在后续实验中选择使用混合的大熊猫粪便样品进行菌群移植;2.造模后小鼠粪便菌群与熊猫粪便菌群的平均相似度显著提高,达到36%以上,显著高于普通小鼠与大熊猫粪便的菌群相似度对比值;3.通过组间比较,多次灌喂的小鼠粪便菌群的组间相似性(70%)显著高于单次灌喂组(低于50%),组间更加稳定;多次灌喂的结果,条带数间的差异显著低于单次灌喂(P<0.05),稳定性明显提升。结论:利用抗生素处理得到的伪无菌小鼠,可以用于建造大熊猫源化的小鼠肠道模型,并且多次灌喂可以使得模型更加稳定。该实验结果将为后续开展更为严谨和深入的大熊猫源化小鼠模型建造提供重要的理论依据。

关键词：熊猫肠道菌群无菌小鼠 PCR-DGGE

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猪源发酵乳杆菌的筛选及其益生特征评价

猪源发酵乳杆菌的筛选及其益生特征评价

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中国畜牧兽医学会动物微生态学分会第五届第十二次全国学术研讨会

作者：王志林王丽蒋宗勇马现永杨雪芬温晓鹿胡胜兰广东省农业科学院动物科学研究所畜禽育种国家重点实验室农业部华南动物营养与饲料重点实验室广东省动物育种与营养公共实验室

饲用抗生素广泛使用造成的药物残留和抗药性等问题已引起广泛关注,减少或禁用饲用抗生素是养殖业面临大趋势。在多种抗生素替代品中,益生菌由于在免疫功能调节、炎症性肠道疾病治疗、肠道应激综合征和腹泻防治等方面发挥重要作用日益受... 详细信息

饲用抗生素广泛使用造成的药物残留和抗药性等问题已引起广泛关注,减少或禁用饲用抗生素是养殖业面临大趋势。在多种抗生素替代品中,益生菌由于在免疫功能调节、炎症性肠道疾病治疗、肠道应激综合征和腹泻防治等方面发挥重要作用日益受到重视。来源于畜禽动物的益生菌更有利于其适应属主特异性肠道环境从而发挥最大功效,本研究旨在从健康仔猪肠道分离一株具有典型益生特征的猪源乳酸杆菌。首先,采取断奶2周后的健康仔猪肛门处采集新鲜粪便,经梯度稀释后均匀涂布到Rogosa SL完全选择性培养基上,挑选出疑似菌株106株,经革兰氏染色镜检,筛选得到90株细胞形态呈杆状的阳性菌株。再经抗逆性(pH2.5和0.3%胆盐)筛选,得到16株能耐酸、耐胆盐的候选菌株。候选菌株经16SrDNA序列测序并进行GenBank序列比对,筛选得到1株发酵乳杆菌(Lactobacillus fermentum)LF16菌株。采用体外评价方法对*** LF16菌株的耐酸耐胆盐能力、抑菌活性以及肠上皮细胞的粘附能力等益生特征进行了研究。将*** LF16菌株接种至pH2.5的MRS酸性培养基和含0.3%胆盐MRS培养基中,37℃培养6h后取样平板计数,考察*** LF16菌株耐酸耐胆盐能力,结果发现*** LF16菌株酸性环境和胆盐条件下存活率分别为36.6%和80.2%。采用抑菌圈法研究了*** LF16菌株对产肠毒素大肠杆菌K88(ETEC)、金黄色葡萄球菌、野生型猪链球菌2型和野生型猪链球菌D型等猪常见致病菌抑制作用,结果显示*** LF16菌株对上述各致病菌的抑菌圈分别为11.2mm、9.9mm、14.1mm和17.6mm,抑菌结果表明*** LF16菌株对常见致病菌均有较强抑菌作用。以肠上皮细胞系IPEC-1细胞作为粘附对象,将浓度为1×10～8 cfu/mL的*** LF16菌株添加到贴壁IPEC-1单层细胞的培养皿中,共孵育2h,清洗除去未粘附的细菌,收集细胞并稀释至合适浓度,采用涂布平板计数法计数粘附到IPEC-1细胞上*** LF16细菌数量,结果发现*** LF16菌株对IPCE-1细胞具有较强的粘附能力。综上所述,经大规模筛选和体外益生特征评价,我们筛选得到一株猪源发酵乳杆菌*** LF16菌株,其在抗逆性、抑菌能力以及粘附能力等方面均表现出典型益生菌的基本特征。

关键词：发酵乳杆菌益生特征体外评价

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Intestinal microbiota and oral administration of Enterococus faecium associated with the growth performance of newborn piglets

Intestinal microbiota and oral administration of Enterococus...

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中国畜牧兽医学会动物微生态学分会第五届第十二次全国学术研讨会

作者： Y.B.Wang W.Du A.K.Fu X.P.Zhang Y.Huang K.H.Lee K.Yu W.F.Li Y.L.Li College of Animal Science Zhejiang University National Engineering Laboratory of Bio Feed Safety and pollution prevention Key Laboratory of Molecular Animal NutritionMinistry of EducationKey Laboratory of Animal Nutrition and Feed Science in East ChinaMOA College of Biosystems Engineering and Food Science Zhejiang University College of Animal Science and Technology Guangxi University Faculty of Medicine Dentistry and Health ScienceThe University of Western Australia

The oral administration of Enterococus faecium(EFl) to newborn suckling and weaned piglets along with their growth performances and intestinal microbiota was investigate in this study..Twenty-four newborn piglets were initially divided into 2 *** treatment or probiotics group receiving 2 ml of 10%sterilized skim milk by oral gavage was supplemented with viable Enterococcus faecium EF1(6×10～8 CFU/mL) at the first,the third and the fifth day after birth,while the control group was made of 2 ml of 10%sterilized skim milk without probiotic at the same *** showed that oral administration of Enterococcus faecium EF1 was associated with a remarkable increaseon the body weight of piglets for both suckling and weaning periods,by 30.73%(P<0.01) and 320.84%(P<0.01),and also decreased the diarrhea rate,by 43.21%(P<0.05) and 71.42%(P<0.05),*** addition,454-pyrosequencing analysis revealed that there was no significant difference in the intestinal microbial diversity of the suckling piglets between the two groups,nevertheless,when compared to the control group,the relative abundance of Firmicutes in the probiotics group substantially augmentedy,while the relative abundance of Proteobacteria,Bacteroidetes and Fusobacteria ***,result indicated that oral administration of Enterococus faecium EF1 did not have any influence on the relative abundance of Firmicutes in weaned piglets rather than increasing the relative abundance of Bacteroidetes and decreasing the relative abundance of ***,at level of Firmicutes phylum,the relative abundance of Lactobacillales in theprobiotic group increased *** findings suggest that oral administration of *** EF1 to newborn piglets could improve the growth performance and intestinal microbiota of piglets for both suckling and weaning periods.

关键词： Enterococus faecium EF1 piglets growth performance microbiota

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Design,Characterization and Expression of a Novel Hybrid Peptide Melittin(1-13)-LL37(17-30)

Design,Characterization and Expression of a Novel Hybrid Pep...

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中国畜牧兽医学会动物微生态学分会第五届第十二次全国学术研讨会

作者： Xubiao Wei Dayong Si Zhaojun Zheng Lulu Zhang Rujuan Wu Rijun Zhang Laboratory of Feed Biotechnology State Key Laboratory of Animal NutritionCollege of Animal Science and TechnologyChina Agricultural University

Both Melittin(ME) and LL37(L) are small linear helical peptides with multiply effects including antibacterial,anti-inflammation,anti-cancer and so on[1-3].However,they also have the undesirable property of cytotoxicity to red blood cells at a lower concentration(2 μM and 25-30 mM) which largely limit application widely in clinic[1-3].Recently,hybridizing of different antimicrobial peptides(AMPs) has been a common practice for obtaining novel hybrid AMPs with elevated antibacterial activity but minimized cytotoxicity[4,5].The hybrid peptide ME(1-13)-L(17-30)(M-L)combining the hydrophobic N-terminal fragment of melittin(M) with the core antibacterial fragment of LL37(L),was designed for the first time to explore its antibacterial activity and hemolytic activity against bacteria and sheep erythrocyte *** showed that M-L had an even more potent antibacterial activity against all indicator strains(especially gram-positive bacteria) than M and L,whereas did not exhibit hemolytic activity to sheep erythrocytes,implying M-L can be served as a potential therapeutic drug to substitute traditional ***,the high expense of biosynthesis limited its further research,therefore fusion expression of M-L was carried out in Escherichia coli(***)for overproducing the hybrid peptide so as to solve the *** DNA sequence encoding M-L with preferred codons was cloned into the pET-SUMO vector for protein expression in *** BL21(DE3).After IPTG induction,approximately 165 mg soluble fusion protein SUMO-M-L was recovered per liter supernatant of the fermentation ultrasonic lysate using Ni-NTA sepharose column(92%purity).And 23 mg recombinant M-L was obtained per liter culture after cleavage of SUMO protease and purification of Ni-NTA sepharose *** sum,this research not only supplied an effective approach for overproducing hybrid peptide M-L,but paved the way for its further exploration on pharmaceutical potential and medical importance.

关键词： Hybrid peptide Melittin(1-13)-LL-37(17-30) Antibacterial activity Hemolytic activity Escherichia coli Fusion expression

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