Refining the gut colonization Zophobas morio larvae model using an oral administration of multidrug-resistant Escherichia coli.

IF 3.7 3区 医学 Q2 INFECTIOUS DISEASES Journal of global antimicrobial resistance Pub Date : 2024-11-02 DOI:10.1016/j.jgar.2024.10.262
Yasmine Eddoubaji, Claudia Aldeia, Dierik H Heg, Edgar I Campos-Madueno, Andrea Endimiani
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Abstract

Background: The darkling beetle Zophobas morio can be implemented as an alternative in vivo model to study different intestinal colonization aspects. Recently, we showed that its larvae can be colonized by multidrug-resistant Escherichia coli strains administered via contaminated food (for 7 days) for a total experimental duration of 28 days.

Method: In the present work, we aimed to shorten the model to 14 days (T14) by administering the previously used CTX-M-15 ESBL-producing ST131 Escherichia coli strain Ec-4901.28 via a single oral administration (5 µL dose of 108 CFU/mL) , using a blunt 26s-gauge needle connected to a 250 μL gastight syringe. Force-feeding was performed either without or with (larvae placed on ice for 10 minutes before injection) anesthesia. In addition, phage-treated larvae were orally injected with 10 µL of INTESTI bacteriophage cocktail (∼105-6 PFU/mL) on days 4 (T4) and 7 (T7) .

Results: Growth curve analyses showed that, while larvae rapidly became colonized with Ec-4901.28 (T1, ∼106-7 CFU/mL) , only those anesthetized maintained a high bacterial load (∼102-3vs. ∼105-6 CFU/mL) and survival rate (76% vs. 99%; P<0.001) by T14. Moreover, bacteriophage administration to anesthetized larvae significantly reduced the bacterial count of INTESTI-susceptible Ec-4901.28 at T14 (5.17 × 105vs. 2.26 × 104, for non-treated and phage-treated larvae, respectively; P=0.04) .

Conclusion: The methodological refinements applied to establish the intestinal colonization model simplify the use of Z. morio larvae, facilitate prompt evaluation of novel decolonization approaches and reduce experiments involving vertebrate animals in accordance with the Replacement, Reduction and Refinement principles.

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利用口服耐多药大肠杆菌改进 Zophobas morio 幼虫肠道定植模型。
背景:暗纹甲虫(Zophobas morio)可作为研究不同肠道定植问题的替代活体模型。最近,我们发现其幼虫可通过受污染的食物(7 天)接受耐多药大肠杆菌菌株的定植,实验总持续时间为 28 天:在本研究中,我们使用 26 号钝针与 250 μL 密封注射器连接,通过单次口服给药(5 μL 剂量,108 CFU/mL)给药先前使用过的 CTX-M-15 ESBL 产 ST131 大肠埃希菌菌株 Ec-4901.28,旨在将模型时间缩短至 14 天(T14)。在不麻醉或麻醉(注射前将幼虫放在冰上 10 分钟)的情况下进行强制喂食。此外,在第 4 天(T4)和第 7 天(T7),给噬菌体处理过的幼虫口服 10 µL INTESTI 噬菌体鸡尾酒(∼105-6 PFU/mL):生长曲线分析表明,虽然幼虫很快就被 Ec-4901.28 定殖(T1,∼106-7 CFU/mL),但只有那些被麻醉的幼虫才能保持较高的细菌量(∼102-3vs. ∼105-6 CFU/mL)和存活率(76% vs. 99%;P5vs:建立肠道定殖模型的方法改进简化了莫氏螯虾幼虫的使用,有利于及时评估新型非殖化方法,并根据 "替换、减少和改进 "原则减少了涉及脊椎动物的实验。
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来源期刊
Journal of global antimicrobial resistance
Journal of global antimicrobial resistance INFECTIOUS DISEASES-PHARMACOLOGY & PHARMACY
CiteScore
8.70
自引率
2.20%
发文量
285
审稿时长
34 weeks
期刊介绍: The Journal of Global Antimicrobial Resistance (JGAR) is a quarterly online journal run by an international Editorial Board that focuses on the global spread of antibiotic-resistant microbes. JGAR is a dedicated journal for all professionals working in research, health care, the environment and animal infection control, aiming to track the resistance threat worldwide and provides a single voice devoted to antimicrobial resistance (AMR). Featuring peer-reviewed and up to date research articles, reviews, short notes and hot topics JGAR covers the key topics related to antibacterial, antiviral, antifungal and antiparasitic resistance.
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