Abstract：Objective To evaluate the pathogenicity of Acinetobacter venetum（Av），which is expected to be used as an environmental remediation agent，using Caenorhabditis elegans（C.elegans）. Methods The C.elegans were cultured on the media loaded with E.coli OP50 and Av，respectively. The pathogenicity of Av was evaluated by observing the effects of Av on the growth，movement，digestive function，lifespan and reproduction of C.elegans，compared with that of another evaluation system according to NY 1109-2017 General Biosafety Standard for Microbial Fertilizers. ResultsBy C. elegans system，it was found that the body length，width，head thrash frequency，body bending frequency and average lifespan [（13.5±0.4）d vs.（13.7±0.4）d] of adult nematodes in the Av group were not significantly different from those in the OP50 group（all P>0.05）；while the average time of defecation cycle in the Av group shortened，the total number of progenies in the Av group increased by 18.7%（all P<0.05）. According to NY1109-2017 General Biosafety Standard for Microbial Fertilizers，it was found that the oral LD50 values for both male and female mice were more than 10 g/kgbw，which was practically non-toxic；the pathogenicity test of acute intraperitoneal injection showed that the animals did not have signs of poisoning，deaths or any abnormalities in gross anatomy；Av had no irritation to damaged skin and eyes of rabbits；the hemolysis test was negative；Av was sensitive to seven antibiotics and was medium to one antibiotic. Conclusion Av is not pathogenic. C. elegans can be used in early screening for the pathogenicity of environmental remediation agents.
[1］ KOTHARI A，CHARRIER M，WU Y W，et al. Transcriptomic analysis of the highly efficient oil-degrading bacterium Acinetobacter venetianus RAG-1 reveals genes important in dodecane uptake and utilization[J/OL]. FEMS Microbiol Lett，2016，363（20）（2016- 09-22）[2019-08-01]. https：//www.ncbi.nlm.nih.gov/pmc/articles/ PMC5074533. [2］ FONDI M，MAIDA I，PERRIN E，et al. Genomic and phenotypic characterization of the species Acinetobacter venetianus[J]. Sci Rep，2016，6：21985. [3］ LUCKARIFT H R，SIZEMORE S R，FARRINGTON K E，et al. Biodegradation of medium chain hydrocarbons by Acinetobacter venetianus 2AW immobilized to hair-based adsorbent mats[J]. Biotechnol Prog，2011，27（6）：1580-1587. [4］ LAMAS B，MICHEL M L，WALDSCHMITT N，et al. Card9 mediates susceptibility to intestinal pathogens through microbiota modulation and control of bacterial virulence[J]. Gut，2018，67（10）：1836-1844. [5］ BOEHNKE K F，EATON K A，FONTAINE C，et al. Reduced infectivity of waterborne viable but nonculturable Helicobacter pylori strain SS1 in mice[J/OL]. Helicobacter，2017，22（4）（2017- 04-24）[2019-08-01]. https：//onlinelibrary.wiley.com/doi/full/10. 1111/hel.12391. [6］ MANN K M，PRIDE A C，FLENTIE K，et al. Analysis of the contribution of MTP and the predicted Flp pilus genes to Mycobacterium tuberculosis pathogenesis[J]. Microbiology，2016， 162（10）：1784-1796. [7］ 中华人民共和国农业部. 微生物肥料生物安全通用技术准则：NY 1109—2017[S]. 北京：中国农业出版社，2018：2-6. [8］ 郭欣欣，于新惠，张颖，等. 单核细胞增生李斯特菌对秀丽隐杆线虫致病性的研究[J]. 微生物学杂志，2015，35（4）：7-12. [9］ 鞠守勇，陈其国. 秀丽隐杆线虫作为病原菌宿主模型的研究概述[J]. 动物医学进展，2018，39（7）：108-113. [10］ 曹玲，陈丽丹，肖斌，等.秀丽隐杆线虫-产碳青霉烯酶阴沟肠杆菌感染模型的建立及应用[J]. 中国抗生素杂志，2018，43（4）：444-449. [11］ BAI Y，ZHI D，LI C，et al. Infection and immune response in the nematode Caenorhabditis elegans elicited by the phytopathogen Xanthomonas[J]. J Microbiol Biotechnol，2014，24（9）：1269- 1279. [12］ VALLEJO J A，BECEIRO A，RUMBO-FEAL S，et al. Optimisation of the Caenorhabditis elegans model for studying the pathogenesis of opportunistic Acinetobacter baumannii[J/OL]. Int J Antimicrob Agents，2015（2015-07-09）[2019-08-01]. http：//www.sciencedirect.com/science/article/pii/S0924857915002411. [13］ HUANG C W，LI S W，HSIU-CHUAN L V. Chronic ZnO-NPs exposure at environmentally relevant concentrations results in metabolic and locomotive toxicities in Caenorhabditis elegans[J]. Environ Pollut，2017，220（Pt B）：1456-1464. [14］ ERMOLAEVA M A，SCHUMACHER B. Insights from the worm：the C. elegans model for innate immunity[J]. Semin Immunol，2014，26（4）：303-309. [15］ HWANG S B，CHOI J G，WEI S，et al. In vivo screening platform for shiga toxin-producing Escherichia coli（STEC）using Caenorhabditis elegans as a model[J]. PLoS One，2018，13（2）：e0193277. [16］ SAMUEL B S，ROWEDDER H，BRAENDLE C，et al. Caenorhabditis elegans responses to bacteria from its natural habitats[J]. Proc Natl Acad Sci USA，2016，113（27）：3941-3949. [17］ LUO H，XIONG J，ZHOU Q，et al. The effects of Bacillus thuringiensis Cry6A on the survival，growth，reproduction，locomotion，and behavioral response of Caenorhabditis elegans[J]. Appl Microbiol Biotechnol，2013，97（23）：10135-10142. [18］张孟丽，束成杰，张卫明. 1-脱氧野尻霉素对秀丽隐杆线虫的抗衰老作用[J]. 食品工业科技，2018，39（21）：280-286.
