Detoxification functionality of lactic acid bacteria

乳酸菌的解毒功能

• 批准号: RGPIN-2019-04028
• 负责人: Reid, Gregor
• 金额: $ 2.58万
• 依托单位: University of Western Ontario
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=714325
• 关键词: Detoxification; functionality; lactic; acid; bacteria

Having shown in our current NSERC Discovery Grant that lactobacilli can perform detoxification of heavy metals, aflatoxins and pesticides, we now wish to identify the mechanisms involved in mitigating the pesticide effects. This is important to counter colony collapse disorder in honey bees. Nothing endangers the global food supply more than the loss of these pollinators. Our preliminary findings suggest that commonly used pesticides impair insect immunity making them more susceptible to a variety of pathogens. Hypothesis We hypothesize that members of the genus Lactobacillus harbor the ability to detoxify pesticides that endanger honey bees and other life forms. Long term goals The long-term goal of this research program is to advance our knowledge of an important bacterial genus with a long history of safe use in foods and supplements, and to exploit attributes that counter environmental chemicals and improve the survival of pollinators. Research objectives 1. 1. Characterize how lactobacilli mitigates the immunosuppressive effects of pesticides and susceptibility to pathogens 2. Determine the functional capacity of lactobacilli to detoxify agriculturally-relevant xenobiotics. Methodological approach We will use Drosophila melanogaster as a well-established and strongly predictive model of the honey bee to determine how lactobacilli affect the co-modulatory response of innate immunity (highly conserved across broad taxonomic range) towards pesticides and/or pathogens as they apply primarily to honeybees, but also other hosts. We will utilize our stocks of RNAi-mediated knockdown and overexpression mutants of the DUOX and Imd pathways, and couple these knockouts with qPCR-based measurement of Imd and DUOX pathway-related gene expression and immunofluorescent stain relevant immune proteins. We will expose flies to different classes of pesticides: neonicotinoids, organophosphates, carbamates, Roundup (glyphosate), acephate, -cyhalothrin, oxamyl, tetraconazole administered at different concentrations. Lactobacillus rhamnosus GR-1, L. plantarum Lp39, L. kunkeei BR1, L. reuteri RC-14, L. rhamnosus GG and strains from African bees will be used to counter the pesticides and determine if they can actually degrade the chemicals. We will compare the microbiota in bees from healthy hives to that of hives infected with P. larvae, Melissococcus plutonius, N. apis, N. ceranae, Ascosphaera apis, Crithidia bombi or Varroa destructor, pathogens that will be examined in vitro for the ability of the lactobacilli to inhibit them. The Drosophila model will also be used to test various lactobacilli for their ability to modulate detoxification pathways. Significance The net result will be to develop an understanding of how lactobacilli strains can counter the most widely used pesticides through their degradation, and by modulation of insect immune responses, microbiota resilience, and countering key honey bee pathogens.

在我们目前的NSERC发现赠款中表明，乳酸杆菌可以对重金属，黄曲霉毒素和农药进行排毒，我们现在希望确定涉及减轻农药作用的机制。这对于蜜蜂中的菌落崩溃障碍很重要。没有什么比失去这些传粉媒介的损失更危及全球粮食供应了。我们的初步发现表明，常用农药会损害昆虫免疫，使其更容易受到各种病原体的影响。 假设 我们假设乳杆菌属的成员具有危害蜜蜂和其他生命形式的农药的能力。 长期目标 该研究计划的长期目标是提高我们对重要细菌属的了解，并具有悠久的食品和补品的安全历史，并利用应对环境化学物质并改善授粉媒介存活的属性。 研究目标 1。1。表征乳酸杆菌如何减轻农药的免疫抑制作用和对病原体的易感性 2。确定乳酸杆菌对农业与农业相关的异种生物的排毒能力。 方法论方法 我们将使用果蝇Melanogaster作为蜂蜜蜜蜂的公认和强烈预测的模型，以确定乳酸杆菌如何影响先天免疫（跨广泛的分类范围高度保守）对农药和/或病原体的共调节反应，因为它们主要适用于蜜蜂，但也适用于蜜蜂。我们将利用DUOX和IMD途径的RNAi介导的敲低和过表达突变体的库存，并将这些基因敲除与基于QPCR的IMD和DUOX途径相关基因表达以及免疫荧光染色的相关免疫蛋白的测量。我们将暴露于不同类别的农药：新烟碱，有机磷酸盐，氨基甲酸酯，综述（草甘膦），屈服， - 环霉素，甲氧氨基蛋白，甲氧苄酮，四氯康唑，以不同的浓度给药。乳杆菌Rhamnosus gr-1，L。Plantarum Lp39，L。KunkeeiBr1，L。ReuteriRC-14，L。Rhamnosus GG和非洲蜜蜂的菌株将用于抵抗农药并确定它们是否可以实际降解化学物质。我们将比较从健康蜂巢中的菌群与感染了幼虫的蜂窝，梅洛氏菌Plutonius，N。Apis，N。Ceranae，Ascosposphaera apis，Crithidia bombi或Varroa Destructor，病原体，将在体外检查lactobacacilli inshiminibali insibnibhi insibhibAcilli insibniby insibhibaCilli insibniby insibhibAcilli in nimighibAcillia。 果蝇模型还将用于测试各种乳杆菌的调节途径的能力。 意义 最终结果将是对乳酸菌菌株如何通过降解以及通过调节昆虫免疫反应，微生物群弹性和对抗关键的蜂蜜蜜蜂病原体来应对最广泛使用的农药的理解。