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Model prediction of inactivation of aeromonas salmonicida grown on poultry in situ by intense pulsed light

Food Science and Human Wellness 2024年第2期13卷 1011-1017页

作者： Jingwen Wang Zhenzhen Ning Yifan Chen Xinglian Xu Huhu Wang Jiangsu Collaborative Innovation Center of Meat Production and Processing Quality and Safety ControlCollege of Food Science and TechnologyNanjing Agricultural UniversityNanjing 210095China

The aim of this study was to evaluate the factors influencing the inactivation effect of intense pulsed light(IPL)on aeromonas salmonicida grown on chicken meat and skin,and to further develop prediction models of *** this work,chicken meat and skin inoculated with meat-borne *** isolates were subjected to IPL treatments under different *** results showed that IPL had obvious bactericidal effect in the chicken skin and thickness groups when the treatment voltage and time were 7 V combined with 5 *** addition,the lethality curves of *** were fitted under IPL conditions of 3.5-7.5 *** comparison of statistical parameters revealed that the Weibull model could best fit the mortality curves and could accurately predict the mortality dynamic of *** grown on chicken *** further a secondary model between the scale factor b and the treatment voltage in Weibull model was established using linear equations,which determined that the secondary model could accurately predict the inactivation of *** study provides a theoretical basis for future prediction models of aeromonas,and also provides new ideas for sterilization approaches of meat-borne aeromonas.

关键词： aeromonas salmonicida Intense pulsed light Inactivation Chicken meat Model

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Immuno-Protective Efficiency of the Bivalent Inactivated Vaccine Against Vibrio scophthalmi and aeromonas salmonicida Infections in Turbot(Scophthalmus maximus L.)

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Journal of Ocean University of China 2023年第4期22卷 1079-1086页

作者： ZHOU Shun ZHENG Xujia DING Yuanyuan SU Lin HUANG Qing XIU Yunji School of Marine Science and Engineering Qingdao Agricultural UniversityQingdao 266109China

Vibrio scophthalmi and aeromonas salmonicida can cause high turbot mortality and huge economic ***,vaccination is the most promising method for preventing communicable *** this study,we used formalin to kill *** and *** cells,and mixed with the mineralized oil adjuvant(Montanide^(TM)ISA 763 AVG)to prepare the bivalent inactivated *** results showed that turbot inoculated with the bivalent inactivated vaccine exhibited strong tolerance to the infection of *** and ***,and no obvious clinical symptoms and pathological changes were *** activities of enzymes lysozyme,acid phosphatase and complement C3 had significantly increased after the *** antibody titer response of vaccinated turbot was greatly boosted,which was positively connected with the immunological impact according to ELISA ***,the expression levels of immune-related genes such as MHC-IIα,MHC-IIβ,CD4,CD8,TNF-αand IL^(-1)βwere up-regulated,demonstrating that it might stimulate humoral and cellular immunological response in *** findings highlight the potential of the bivalent inactivated vaccine for controlling *** and *** infections in turbot.

关键词： aeromonas salmonicida Vibrio scophthalmi bivalent inactivated vaccine immune response

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A recombinase polymerase amplification with lateral flow assay for rapid on-the-spot detection of aeromonas salmonicida

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Water Biology and Security 2024年第3期3卷 58-64页

作者： Sijie Zhang Yuanxing Zhang Qin Liu Qiyao Wang Yibei Zhang State Key Laboratory of Bioreactor Engineering Laboratory for Aquatic Animal DiseasesEast China University of Science and TechnologyShanghai200237China Southern Marine Science and Engineering Guangdong Laboratory(Zhuhai) Zhuhai519000China Shanghai Engineering Research Center of Maricultured Animal Vaccines Shanghai200237China

aeromonas salmonicida is a common pathogen of salmonid fishes that poses a significant threat to the fresh water and marine culture industry,potentially resulting in huge economic *** prevent and control fish diseases caused by ***,rapid and effective diagnostic approaches must be developed,and which are important for routine monitoring and clinical *** combining recombinase polymerase amplification(RPA)technology with a visible lateral flow strip(RPA-LF),we have enhanced both the precision of RPA detection and the convenience of real-time *** this study,we introduce a robust method for detecting *** using *** assay specifically targets the ASA_1441 gene of ***,ensuring high specificity,without cross-reactivity with other prevalent fresh water or marine *** optimal amplification temperature of the RPA assay was 39℃.Its sensitivity extends to as low as 100 fg of purified DNA,representing more than 1000-fold higher sensitivity than conventional PCR ***,to enhance the usability of the RPA-LF assay,we developed a rapid sample preparation method using cellulose dipsticks for nucleic acid *** method achieves a limit of detection(LOD)as low as 1.67 CFU/μL and completes the entire process within 20 *** conclusion,our findings present a rapid and precise tool for monitoring *** infection in aquaculture and marine *** advancement offers valuable insights for effective disease prevention and control strategies.

关键词： aeromonas salmonicida Recombinase Polymerase Amplification(RPA) Lateral flow On-the-spot detection

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A New Furunculosis Challenge Method for Evaluation of Vaccine Efficacy in Rainbow Trout

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Open Journal of Immunology 2016年第4期6卷 136-147页

作者： Moonika H. Marana Jiwan K. Chettri Jakob Skov Inger Dalsgaard Per W. Kania Kurt Buchmann Department of Veterinary Disease Biology Faculty of Health and Medical Sciences University of Copenhagen Frederiksberg Denmark National Veterinary Institute Technical University of Denmark Frederiksberg Denmark

Experimental infection of fish for vaccine efficacy studies is associated with several limitations. Administration of live bacteria with the purpose of causing disease in fish can be performed by co-habitation, immersion or injection. We have developed a new aeromonas salmonicida challenge method for rainbow trout and have applied it for evaluation of furunculosis vaccine efficacy. The method reveals development of systemic immunity in fish as live bacteria are introduced in the tail fin epidermis distant from the vaccine injection site (peritoneal cavity). This method seeks to mimic natural infection in fish farms where tail biting and therefore bacterial exposure to tail fin ulcers is widespread. By use of a multi-needle device ten epidermal perforations were introduced in the dorsal part of the tail fin of anaesthetized rainbow trout (vaccinated or naive). Subsequently 100 μL (3.4 × 108 colony-forming units (CFU) mL-1) of a 48 hour culture of aeromonas salmonicida subsp. salmonicida strain 090710-1/23 was placed at the perforation site for 60 sec whereafter fish were allowed to regain consciousness in clean freshwater. Immunohistochemistry and scanning electron microscopy illustrated the spread of bacteria from the injection site. Classical furunculosis symptoms associated with a high morbidity rate were observed in control fish whereas vaccinated fish exhibited a significantly higher survival.

关键词： aeromonas salmonicida Challenge Method Furunculosis Immunization Salmonids

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群体感应AHLs信号分子对鱼源杀鲑气单胞菌致腐及抗胁迫能力的影响

群体感应AHLs信号分子对鱼源杀鲑气单胞菌致腐及抗胁迫能力的影响

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作者：刘丽浙江工商大学

学位级别：硕士

群体感应(quorum sensing,QS)参与调控食源性微生物的致病性和致腐性。气单胞菌为气单胞菌属(Aeramonas)的革兰氏阴性直杆菌,是水产养殖业重要的条件致病菌和生鲜鱼类贮藏中的腐败菌。前期研究已从腐败大黄鱼中分离出致腐性强的杀鲑气... 详细信息

群体感应(quorum sensing,QS)参与调控食源性微生物的致病性和致腐性。气单胞菌为气单胞菌属(Aeramonas)的革兰氏阴性直杆菌,是水产养殖业重要的条件致病菌和生鲜鱼类贮藏中的腐败菌。前期研究已从腐败大黄鱼中分离出致腐性强的杀鲑气单胞菌(aeromonas salmonicida),发现***能够诱导群体感应生物报告菌产生颜色,提示该细菌具有N-酰基高丝氨酸内酯(N-acylhomoserine lactone,AHLs)活性。然而,鱼源致腐菌salmonicida产生的AHLs与致腐内在调控作用研究仍较少。鉴于此,本论文以*** AE03为研究对象,分析在不同培养条件下AHLs的形成规律和种类;扩增并分析编码AHLs的基因asaI,构建asaI基因突变株;比较研究野生株、asaI突变株和C4-HSL回补株在生长、腐败表型、运动性、生物被膜形成和抗胁迫性的影响。进一步采用qRT-PCR技术探究asaI缺失对致腐和抗胁迫性相关基因表达的影响。研究旨在揭示AHLs介导的QS对*** 致腐和抗胁迫能力的调控作用,为食品腐败菌机理和控制研究提供理论支持。主要结果如下:利用生物报菌紫色杆菌(Chromobacterium violaceum)CV026检测了*** AE03不同培养条件下AHLs的形成规律。结果发现4℃和28℃下培养的AE03均能使CV026产生紫色反应,其中在4℃培养5d和28℃培养30h的上清诱导产生的紫色圈最大,呈现密度依赖性。随着培养基中氯化钠浓度增高,AE03产生的AHLs活性逐步增强,其中1.5%氯化钠紫色圈最大,之后紫色圈减少。采用高效液相色谱串联质谱(HPLC-MS/MS)检测培养上清中AHLs种类和含量,显示 AE03 能够产生 C4-HSL、C6-HSL、C8-HSL、C10-HSL、C12-HSL五种信号分子,其中C4-HSL含量最高,并且28℃培养30 h达到最高,与报告菌结果一致。AE03上清在弱酸或强碱处理下AHLs活性明显下降,其中pH 7.8时AHLs活性保持最高。高温加热能降解菌体上清中AHLs信号分子,随着加热时间的延长,CV026变色圈直径迅速减少,AHLs活性逐渐丧失。采用PCR在*** AE03中扩增出asaI基因,通过融合PCR和自杀性质粒,构建*** AE03 asaI基因突变株。AHLs活性检测显示,突变株asaI丧失诱导报告菌CV026产生紫色,HPLC-MS/MS也发现AHLs含量很低,未检测到C4-HSL和C6-HSL信号分子。比较研究野生株、asaI突变株及C4-HSL回补株在体外培养和鱼汁中的腐败表型、生物被膜、抗胁迫性和致腐能力的差异。研究显示,AE03野生株、asaI突变株和C4-HSL回补株在28℃下生长无显著差异。相比于野生株,asaI突变株在12 h、18 h和24 h后三甲胺(TMA)分别增加100.2%、22.3%和24.9%,而添加C4-HSL回补株TMA有所下降,特别是添加40μMC4-HSL时TMA分别下降52.7%、28.5%和27%。相似地,asaI突变株在培养12 h、18 h、24 h形成的生物胺比野生株分别增加 21.5%、22.4%和 23.2%(p<0.05),而 20 μM C4-HSL回补株分别下降 16.3%、7.2%和 6.5%(p<0.05),40 μM C4-HSL 处理组则减少28.6%,14.5%和16.4%(p<0.05)。突变株胞外蛋白酶的活性也显著提高(p<0.05),外源添加4-HSL导致突变株分泌胞外蛋白酶活性下降,与野生株的蛋白酶活性接近。可见与野生株相比,asaI突变株腐败表型都显著增强,并且外源C4-HSL添加浓度越高,致腐产物形成降低,表现浓度依赖性。结晶紫染色法发现*** AE03生物被膜形成速度较快,其中培养12 h被膜产量达到最大值,而asaI突变株比野生株的被膜量减少32.1%,共聚焦显微镜观察也显示AE03成熟的生物被膜厚度为52.52 μm,而asaI突变株仅为36.66 μm。野生株和突变株的抗胁迫性比较研究发现,在50℃热处理后菌体的存活率逐渐减少,其中60 min后野生株和突变株存活率分别下降为46.8%和54.1%(p<0.05);2 mM双氧水处理导致野生株存活率急剧下降,而asaI突变株减少较为缓慢,经5 min处理后两菌株的存活率分别为70.3%和87.1%(p<0.05);30%氯化钠处理90 min时,野生株和突变株存活率分别为50.5%和59.8%(p<0.05)。结果发现,asaaI突变株能够增强菌体对高温、双氧水

关键词： aeromonas salmonicida AHLs asaI 腐败生物被膜抗胁迫性

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