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大肠杆菌O157:H7在被该病原体污染并经化学或微生物添加剂处理的玉米青贮饲料中的命运及细菌多样性

Fate of Escherichia coli O157:H7 and bacterial diversity in corn silage contaminated with the pathogen and treated with chemical or microbial additives.

作者信息

Ogunade I M, Jiang Y, Kim D H, Cervantes A A Pech, Arriola K G, Vyas D, Weinberg Z G, Jeong K C, Adesogan A T

机构信息

Department of Animal Sciences, Institute of Food and Agricultural Sciences, University of Florida, Gainesville 32608.

Division of Applied Life Science (BK21Plus, Institute of Agriculture and Life Science), Gyeongsang National University, Jinju 660-701, South Korea.

出版信息

J Dairy Sci. 2017 Mar;100(3):1780-1794. doi: 10.3168/jds.2016-11745. Epub 2016 Dec 29.

DOI:10.3168/jds.2016-11745
PMID:28041727
Abstract

Inhibiting the growth of Escherichia coli O157:H7 (EC) in feeds may prevent the transmission or cycling of the pathogen on farms. The first objective of this study was to examine if addition of propionic acid or microbial inoculants would inhibit the growth of EC during ensiling, at silo opening, or after aerobic exposure. The second objective was to examine how additives affected the bacterial community composition in corn silage. Corn forage was harvested at approximately 35% dry matter, chopped to a theoretical length of cut of 10 mm, and ensiled after treatment with one of the following: (1) distilled water (control); (2) 1 × 10 cfu/g of EC (ECCH); (3) EC and 1 × 10 cfu/g of Lactobacillus plantarum (ECLP); (4) EC and 1 × 10 cfu/g of Lactobacillus buchneri (ECLB); and (5) EC and 2.2 g/kg (fresh weight basis) of propionic acid, containing 99.5% of the acid (ECA). Each treatment was ensiled in quadruplicate in laboratory silos for 0, 3, 7, and 120 d and analyzed for EC, pH, and organic acids. Samples from d 0 and 120 were also analyzed for chemical composition. Furthermore, samples from d 120 were analyzed for ammonia N, yeasts and molds, lactic acid bacteria, bacterial community composition, and aerobic stability. The pH of silages from all treatments decreased below 4 within 3 d of ensiling. Escherichia coli O157:H7 counts were below the detection limit in all silages after 7 d of ensiling. Treatment with L. buchneri and propionic acid resulted in fewer yeasts and greater aerobic stability compared with control, ECCH, and ECLP silages. Compared with the control, the diversity analysis revealed a less diverse bacterial community in the ECLP silage and greater abundance of Lactobacillus in the ECLP and ECA silages. The ECLB silage also contained greater abundance of Acinetobacter and Weissella than other silages. Subsamples of silages were reinoculated with 5 × 10 cfu/g of EC either immediately after silo opening or after 168 h of aerobic exposure, and EC were enumerated after 6 or 24 h, respectively. All silages reinoculated with EC immediately after silo opening (120 h) had similar low pH values (<4.0) and EC counts were below the detection limit. The ECCH and ECLP silages reinoculated with EC after 168 h of aerobic exposure had relatively high pH values (>5.0) and EC counts (5.39 and 5.30 log cfu/g, respectively) 24 h later. However, those treated with L. buchneri or propionic acid had lower pH values (4.24 or 3.96, respectively) and lower EC counts (1.32 log cfu/g or none, respectively). During ensiling, EC was eliminated from all silages at pH below 4.0. During aerobic exposure, the growth of EC was reduced or prevented in silages that had been treated with L. buchneri or propionic acid at ensiling, respectively.

摘要

抑制饲料中大肠杆菌O157:H7(EC)的生长可能会阻止该病原体在农场中的传播或循环。本研究的首要目标是检验添加丙酸或微生物接种剂是否会在青贮过程中、开窖时或有氧暴露后抑制EC的生长。第二个目标是研究添加剂如何影响玉米青贮饲料中的细菌群落组成。玉米青贮原料在干物质含量约为35%时收割,切碎至理论切割长度10毫米,并在经过以下处理之一后进行青贮:(1)蒸馏水(对照);(2)1×10 cfu/g的EC(ECCH);(3)EC和1×10 cfu/g的植物乳杆菌(ECLP);(4)EC和1×10 cfu/g的布氏乳杆菌(ECLB);以及(5)EC和2.2 g/kg(鲜重基础)的丙酸,其中酸含量为99.5%(ECA)。每个处理在实验室青贮窖中重复青贮4次,分别青贮0、3、7和120天,并分析其中的EC、pH值和有机酸。对第0天和120天的样品还进行了化学成分分析。此外,对第120天的样品分析了氨态氮、酵母和霉菌、乳酸菌、细菌群落组成以及有氧稳定性。所有处理的青贮饲料在青贮3天内pH值均降至4以下。青贮7天后,所有青贮饲料中的大肠杆菌O157:H7数量均低于检测限。与对照、ECCH和ECLP青贮饲料相比,布氏乳杆菌和丙酸处理使酵母数量减少,有氧稳定性提高。与对照相比,多样性分析显示ECLP青贮饲料中的细菌群落多样性较低,而ECLP和ECA青贮饲料中乳酸菌的丰度更高。ECLB青贮饲料中不动杆菌属和魏斯氏菌属的丰度也高于其他青贮饲料。青贮饲料的子样本在开窖后立即或有氧暴露168小时后分别重新接种5×10 cfu/g的EC,然后分别在6小时或24小时后对EC进行计数。所有在开窖后立即(120小时)重新接种EC的青贮饲料pH值均较低(<4.0),EC数量低于检测限。有氧暴露168小时后重新接种EC的ECCH和ECLP青贮饲料24小时后的pH值相对较高(>5.0),EC数量分别为5.39和5.30 log cfu/g。然而,用布氏乳杆菌或丙酸处理的青贮饲料pH值较低(分别为4.24或3.96),EC数量较少(分别为1.32 log cfu/g或无)。在青贮过程中,pH值低于4.0时所有青贮饲料中的EC均被消除。在有氧暴露期间,青贮时用布氏乳杆菌或丙酸处理过的青贮饲料中EC的生长分别受到抑制或阻止。

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