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基于抗菌、抗病毒、免疫调节和抗生物膜活性筛选和选择用于抗牛呼吸道病原体的鼻内喷雾剂的精油。

Screening and selection of essential oils for an intranasal spray against bovine respiratory pathogens based on antimicrobial, antiviral, immunomodulatory, and antibiofilm activities.

作者信息

Amat Samat, Magossi Gabriela, Rakibuzzaman Agm, Holman Devin B, Schmidt Kaycie N, Kosel Luke, Ramamoorthy Sheela

机构信息

Department of Microbiological Sciences, North Dakota State University, Fargo, ND, United States.

Lacombe Research and Development Centre, Agriculture and Agri-Food Canada, Lacombe, AB, Canada.

出版信息

Front Vet Sci. 2024 Feb 7;11:1360398. doi: 10.3389/fvets.2024.1360398. eCollection 2024.

Abstract

INTRODUCTION

The rise in antibiotic resistant pathogens associated with bovine respiratory disease (BRD) poses a serious challenge, particularly to the beef feedlot industry, as they currently depend on antibiotics to prevent BRD to mitigate the financial burden (approx. $1 billion annual loss) inflicted by BRD-associated high mortality and morbidity in feedlot cattle. Thus, there is an impetus need for the development of antimicrobial alternative strategies against BRD. This study aimed to screen and select candidate essential oils (EOs) for the development of an intranasal EO spray that can inhibit BRD pathogens and promote microbiota-mediated respiratory health.

METHODS

The effects of selected EOs (ajowan, cinnamon leaf, citronella, grapefruit, fennel, and thyme) on a bovine nasopharyngeal microbiota culture were evaluated using 16S rRNA gene sequencing. The microbiota culture was enriched by incubating nasopharyngeal swabs obtained from finishing beef heifers in brain heart infusion broth with and without EOs (0.025%, v/v). These EOs were then also evaluated for their immunomodulatory effects on bovine turbinate (BT) cells by analyzing the concentrations of 15 cytokines and chemokines in cell culture after 24 h incubation. The crystal violet assay was done to assess the antibiofilm activity of EOs against UMN026 strain. Finally, 15 EOs were screened for their antiviral activity against the bovine viral diarrhea virus 1 (BVDV-1) using BT cells and a fluorescence-based method.

RESULTS

Ajowan, fennel, and thyme resulted in a moderate reduction of overall nasopharyngeal microbiota growth with significant alterations of both alpha and beta diversity, and the relative abundance of predominant bacterial families (e.g., increasing and decreasing ) compared to the control ( < 0.05). Co-incubation of BT cells with selected EOs resulted in minimal alterations in cytokine and chemokine levels ( > 0.05). Ajowan, thyme, fennel, and cinnamon leaf exhibited antibiofilm activity at concentrations of 0.025 and 0.05%. Reduction of BVDV-1 replication in BT cells was observed with thyme (strong), and ajowan and citronella (moderate) at 0.0125% concentration.

DISCUSSION

Accordingly, ajowan, thyme, fennel, cinnamon leaf, and citronella EOs were selected for further development as an intranasal EO spray to prevent and control of BRD pathogens in feedlot cattle.

摘要

引言

与牛呼吸道疾病(BRD)相关的抗生素耐药性病原体的增加构成了严峻挑战,尤其是对肉牛饲养场行业而言,因为他们目前依赖抗生素来预防BRD,以减轻BRD相关的高死亡率和发病率给饲养场牛造成的经济负担(每年约10亿美元损失)。因此,迫切需要开发针对BRD的抗菌替代策略。本研究旨在筛选和选择候选精油(EOs),以开发一种鼻内EO喷雾剂,该喷雾剂可抑制BRD病原体并促进微生物群介导的呼吸道健康。

方法

使用16S rRNA基因测序评估所选精油(阿育吠陀、肉桂叶、香茅、葡萄柚、茴香和百里香)对牛鼻咽微生物群培养物的影响。通过将从育肥牛小母牛获得的鼻咽拭子在含有和不含有EOs(0.025%,v/v)的脑心浸液肉汤中孵育来富集微生物群培养物。然后,通过分析孵育24小时后细胞培养物中15种细胞因子和趋化因子的浓度,评估这些EOs对牛鼻甲(BT)细胞的免疫调节作用。进行结晶紫试验以评估EOs对UMN026菌株的抗生物膜活性。最后,使用BT细胞和基于荧光的方法筛选15种EOs对牛病毒性腹泻病毒1(BVDV-1)的抗病毒活性。

结果

与对照相比,阿育吠陀、茴香和百里香导致总体鼻咽微生物群生长适度减少,α和β多样性以及主要细菌家族的相对丰度均有显著变化(例如,增加和减少)(P < 0.05)。BT细胞与所选EOs共孵育导致细胞因子和趋化因子水平变化最小(P > 0.05)。阿育吠陀、百里香、茴香和肉桂叶在0.025%和0.05%浓度下表现出抗生物膜活性。在0.0125%浓度下,观察到百里香(强)、阿育吠陀和香茅(中度)可减少BT细胞中BVDV-1的复制。

讨论

因此,选择阿育吠陀、百里香、茴香、肉桂叶和香茅EOs进行进一步开发,作为一种鼻内EO喷雾剂,用于预防和控制饲养场牛中的BRD病原体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/51ca/10879409/aa143e62cb2a/fvets-11-1360398-g001.jpg

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