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变性梯度凝胶电泳-聚合酶链反应比较壳聚糖对肉鸡盲肠细菌和鼠伤寒沙门氏菌厌氧培养的影响。

Denaturing Gradient Gel Electrophoresis-Polymerase Chain Reaction Comparison of Chitosan Effects on Anaerobic Cultures of Broiler Cecal Bacteria and Salmonella Typhimurium.

机构信息

1 Food and Feed Safety Research Unit, Southern Plains Area Research Center , Agriculture Research Service, United States Department of Agriculture, College Station, Texas.

2 Department of Physiology, Government College University , Faisalabad, Pakistan .

出版信息

Foodborne Pathog Dis. 2018 Apr;15(4):246-252. doi: 10.1089/fpd.2017.2365. Epub 2018 Jan 10.

Abstract

Enteropathogen colonization and product contamination are major poultry industry problems. The emergence of antibiotic resistance, and associated risks to human health, is limiting the use of antibiotics as first-line defense against enteropathogens in poultry. The chitin derivative, chitosan, has drawn substantial attention for its bactericidal properties. Different molecular weight (MW) chitosans can have varied effects against different bacteria in monoculture. In the current study, cecal contents from each of three market-age broilers and Salmonella Typhimurium, as indicator enteropathogen, were exposed to in vitro anaerobic culture to three chitosan preparations (0.08%, wt/vol), low (LMW), medium (MMW), and coarse (CMW). Effects of chitosan and the carrier solvent acetic acid, on cecal bacteria and Salmonella, were examined by denaturing gradient gel electrophoresis (DGGE) and Salmonella enumeration. Bacterial profiles for the three cecal contents were shown by DGGE to be very different. Each of the three cecal contents grown in the presence of 0.08% acetic acid was very different from the same contents grown without the chitosan solvent. Culturing cecal contents in the presence of chitosan altered the bacterial DGGE profiles from the control and acetic acid-only cultures. The DGGE chitosan-treated profiles for all three cecal sources were identical to each other regardless of the MW chitosan in the culture medium. Compared with Salmonella in monoculture, Salmonella decreased (p < 0.05) by about 1.5 log CFU/mL when grown in mixed culture with cecal contents. Salmonella monocultures in the presence of 0.08% of the chitosan solvent acetic acid decreased (p < 0.05) counts by almost 3.5 log CFU/mL. Combining acetic acid and cecal contents reduced (p < 0.05) Salmonella by 7 log CFU/mL. Adding the chitosan preparations to the mixtures reduced (p < 0.05) Salmonella by 8 log CFU/mL.

摘要

肠病原体定植和产品污染是家禽业的主要问题。抗生素耐药性的出现及其对人类健康的相关风险,限制了抗生素作为家禽中肠病原体一线防御的使用。壳聚糖是一种甲壳素衍生物,因其杀菌特性而受到广泛关注。不同分子量 (MW) 的壳聚糖在单一培养物中对不同细菌可能有不同的作用。在本研究中,从三只市场年龄的肉鸡的盲肠内容物和肠炎沙门氏菌(作为指示性肠病原体)中,在体外厌氧培养中暴露于三种壳聚糖制剂(0.08%,重量/体积)、低分子量 (LMW)、中分子量 (MMW) 和粗分子量 (CMW)。壳聚糖和载体溶剂醋酸对盲肠细菌和肠炎沙门氏菌的影响通过变性梯度凝胶电泳 (DGGE) 和肠炎沙门氏菌计数进行了检查。DGGE 显示,三种盲肠内容物的细菌图谱差异很大。在 0.08%醋酸存在下培养的三种盲肠内容物与没有壳聚糖溶剂的相同内容物非常不同。在壳聚糖存在下培养盲肠内容物改变了对照和仅含醋酸的培养物中的细菌 DGGE 图谱。无论培养基中的 MW 壳聚糖如何,所有三种盲肠来源的 DGGE 壳聚糖处理图谱彼此相同。与单一培养的肠炎沙门氏菌相比,当与盲肠内容物混合培养时,肠炎沙门氏菌减少(p<0.05)约 1.5 log CFU/mL。在含有 0.08%壳聚糖溶剂醋酸的肠炎沙门氏菌单一培养物中,细菌数量减少(p<0.05)近 3.5 log CFU/mL。将醋酸和盲肠内容物结合使用可使肠炎沙门氏菌减少(p<0.05)7 log CFU/mL。将壳聚糖制剂添加到混合物中可使肠炎沙门氏菌减少(p<0.05)8 log CFU/mL。

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