文章摘要
何夙旭,张美超,张宇婷,徐俐,杨雅麟,李青,周志刚.高丝氨酸内酯酶AI-96对嗜水气单胞菌NJ-1浸浴攻毒斑马鱼保护效应的研究[J].水产学报,2013,37(1):94~100
高丝氨酸内酯酶AI-96对嗜水气单胞菌NJ-1浸浴攻毒斑马鱼保护效应的研究
Effect of Quorum quenching enzyme from Bacillus sp AI-96 on Aeromonas hydrophila NJ-1 bath challenge in zebrafish
投稿时间:2012-01-16  修订日期:2012-10-18
DOI:DOI:10.3724/SP.J.1231.2013.37951
中文关键词: N酰基高丝氨酸内酯酶  嗜水气单胞菌  浸浴攻毒  实时荧光定量PCR
英文关键词: N-acylhomoserine lactonase  Aeromonas hydrophila  bath challenge  Real time PCR
基金项目:科技部农业科技成果转化资金项目(2010GB23260591); 天津市农业科技成果转化与推广项目(201004040); 中国科学院水生生物研究所淡水生态与生物技术国家重点实验室开放基金(2011FB05)资助
作者单位E-mail
何夙旭 中国农业科学院饲料研究所农业部饲料生物技术重点开放实验室 hesuxu@caas.net.cn 
张美超 中国农业科学院饲料研究所农业部饲料生物技术重点开放实验室  
张宇婷 中国农业科学院饲料研究所农业部饲料生物技术重点开放实验室  
徐俐 中国农业科学院饲料研究所农业部饲料生物技术重点开放实验室  
杨雅麟 中国农业科学院饲料研究所农业部饲料生物技术重点开放实验室  
李青 中国农业科学院饲料研究所农业部饲料生物技术重点开放实验室  
周志刚 中国农业科学院饲料研究所农业部饲料生物技术重点开放实验室 zhou_zg@msn.com 
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中文摘要:
      本实验旨在评价斑马鱼口服高丝氨酸内脂酶AI-96对嗜水气单胞菌NJ-1浸浴攻毒的保护效应。实验设置基础料与实验料两组饲料,实验料是在基础料中按3 U/g饲料添加N酰基高丝氨酸内酯酶AI-96(以下简称AI-96),通过高浓度(2.5×10 8 cfu/mL)及低浓度(0.7×10 8 cfu/mL)两组剂量的嗜水气单胞菌 NJ-1(以下简称NJ-1)分别浸浴攻毒斑马鱼,在12h、24h、3d、7d和14d取鳃丝、肠道壁、肝和肾样,采用实时荧光定量PCR法检测取样器官中NJ-1量,并统计攻毒周期内的死亡率,来评价高丝氨酸内酯酶AI-96的保护力。试验结果表明在攻毒周期内所取组织内均检测到NJ-1,按菌数肠>鳃>肝>肾,其中高NJ-1剂量未加酶组各组织NJ-1数量均分别明显高于低剂量处理组。在高剂量攻毒条件下,加酶组各组织NJ-1数量均显著低于未加酶处理组(P < 0.05),鳃除外;在低剂量攻毒条件下,未加酶组的NJ-1数量在鳃(3d)、肠(0.5、1、3、7及14 d)、肝(3d)和肾(7d)显著高于加酶组(P < 0.05),其余差异不显著(P > 0.05)。此外无论在高、低剂量攻毒条件下,加酶组的死亡率均低于未加酶组,其中低剂量攻毒7d及以后其死亡率显著低于对照(P < 0.05)。因此口服AI-96可以有效预防NJ-1 ≤0.7×108 cfu/mL范围内的侵袭。
英文摘要:
      The aim of the present experiment was to study the effects of quorum quenching enzyme AI-96 against Aeromonas hydrophila NJ-1 attack with the bath challenge in zebrafish. The experiments set two dietary groups for zebrafish,1) basal diet as control, 2) enzyme-containing diet with 3 U/g food of AI-96. Zebrafish tissues (gill, liver, kidney and intestine) were sampled at day 0.5, 1, 3, 7 and 14 after co-bath challenge with live Aeromonas hydrophila NJ-1 at two doses (high 2.5×10 8 cfu/mL, low 0.7×10 8 cfu/mL). The populations of NJ-1 colonizing in the tissues were quantified by real time PCR, and accumulated mortality of zebrafish were recorded everyday. Results showed that after challenge, the NJ-1 were detectable in the tissues of the attacked zebrafish in the total population order of intestine> gill > liver > kidney. More NJ-1 colonized in the tissues in the high dosage challenge treatment than the low group. Interestingly, the diets containing AI-96 enzmye significantly decreased the colonizing populations of NJ-1 in the zebrafish (P < 0.05), compared with the counterparts fed with basal diets when the fish was chanllenged with 2.5×108 cfu/mL NJ-1. In 0.7×10 8 cfu/mL challenge treatment, the NJ-1 number colonizing in gill (3d), intestine (0.5, 1, 3, 7, 14 d), liver (3 d) and kidney (7 d) were also lower in the fish treated with AI-96 containing diets than the ones with basal diets (P < 0.05). Moreover, these effect was more obviously in 0.7×10 8 cfu/mL challenge treatment after 7 d (P < 0.05). Therefore, the dietary AI-96 enzyme could improved the survival rate against NJ-1 attack in both challenge levels, For this reason, quorum quenching enzyme could effectively protect zebrafish against A. hydrophila at 0.7×10 8 cfu/mL level in zebrafish.
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