饲喂不同抗生素对肉牛废弃粪便中四环素耐药基因数量及持久性的影响

›› 2012, Vol. 39 ›› Issue (10): 101-105.

饲喂不同抗生素对肉牛废弃粪便中四环素耐药基因数量及持久性的影响

金鑫, 张文广, 张燕军, 苏蕊, 王瑞军, 李金泉

内蒙古农业大学动物科技学院,内蒙古呼和浩特 010018

收稿日期:2012-03-15 出版日期:2012-10-20 发布日期:2012-10-19
通讯作者: 李金泉(1957-),男,内蒙古人,博士生导师,教授。E-mail:lijinquan_nd@126.com E-mail:lijinquan_nd@126.com
作者简介:金鑫(1983-),女,内蒙古人,博士生,研究方向:微生物耐药性。

Abundance and Persistence of Tetracycline Resistance Genes in Fecal Depositsy from Beef Cattle Fed Different Subtherapeutic Antibiotics

JIN Xin, ZHANG Wen-guang, ZHANG Yan-jun, SU Rui, WANG Rui-jun, LI Jin-quan

College of Animal Science, Inner Mongolia Agricultural University,Huhhot 010018,China

Received:2012-03-15 Online:2012-10-20 Published:2012-10-19

摘要/Abstract

摘要： 通过对肉牛饲喂低于治疗水平的不同种类抗生素,研究其对肉牛废弃粪便微生物群落中四环素耐药基因数量与持久性的影响。试验肉牛被分成不同抗生素处理组,即氯四环素组、氯四环素和磺胺甲嘧啶组及对照组。将每个围栏中所有动物的新鲜粪便混匀为一份混合样品作为模式样品(每个处理组3份),分别露天放置,在第7、14、28、42、56、70、84、98、112、126和175 天时采样并提取DNA,利用Real-time PCR方法测定四环素耐药基因tet(B),tet(C),tet(L),tet(M),tet(W)及16S rRNA的浓度。结果显示,16S rRNA的浓度在不同处理间相似,在56 d内均有增加的趋势(P＜0.05);总体上看,四环素组的耐药基因初始浓度较高(P＜0.05);所有处理组的tet(B)和tet(C)浓度到56 d时均增长了1~2个对数级,到175 d时又降低到初始水平,而tet(M)与tet(W)的浓度与其他耐药基因相比较高。因此,四环素耐药基因可以在废弃粪便中持续存在超过175 d,而某些基因的最初数量可能会导致错估其后的变化,其浓度的暂时改变并不能归因于微生物群落数量的变化。

关键词: 肉牛粪便; 四环素; 耐药性基因

Abstract: This study was investigated the effects of administering beef cattle antimicrobials at subtherapeutic concentrations on the abundance and persistence of tetracycline resistance genes within the whole microbial community of fecal waste.Cattlwere administered chlortetracycline, chlortetracycline plus sulfamethazine and no antimicrobials. Model fecal deposits (n=3) were prepared by mixing fresh feces from pens into a single composite sample. Real-time PCR was used to measure concentrations of tetracycline resistance genes tet(B), tet(C), tet(L), tet(M), tet(W) and 16S rRNA in DNA extracted from composite feces after 7, 14, 28, 42, 56, 70, 84, 98, 112, 126 and 175 days in the field. The concentrations of 16S rRNA in feces were similar across treatments and increased by day 56. Generally, initial concentrations of tetracycline resistance genes were greater (P＜0.05) in fecal pats from animals fed chlortetracycline. For all fecal treatments, tet(B) and tet(C) increased 1 to 2 log units by 56 d, and then decreased to the initial 7 d levels by 175 d (P＞0.05). The concentrations of tet(M) and tet(W) were greater than other tetracycline resistance determinants. Tetracycline resistance genes could persist in fecal waste from cattle beyond 175 days and the initial load of some genes might underestimate concentrations at later time points. Temporal changes in the concentrations of resistance genes were likely due to shifts in microbial populations.

Key words: cattle fecal deposits; tetracycline; antimicrobial resistance gene

中图分类号:

S816.73

金鑫, 张文广, 张燕军, 苏蕊, 王瑞军, 李金泉. 饲喂不同抗生素对肉牛废弃粪便中四环素耐药基因数量及持久性的影响[J]. , 2012, 39(10): 101-105.

JIN Xin, ZHANG Wen-guang, ZHANG Yan-jun, SU Rui, WANG Rui-jun, LI Jin-quan. Abundance and Persistence of Tetracycline Resistance Genes in Fecal Depositsy from Beef Cattle Fed Different Subtherapeutic Antibiotics[J]. China Animal Husbandry & Veterinary Medicine, 2012, 39(10): 101-105.

参考文献

1 金鑫,苏蕊,张文广,等. 3种四环素耐药基因型肉牛肠道大肠杆菌对四环素类抗生素的耐药性分析[J].中国畜牧兽医,2012,39(5):187~191.
2 Alexander T W, Reuter T,Sharma R,et al. Longitudinal characterization of resistant Escherichia coli in fecal deposits from cattle fed subtherapeutic levels of antimicrobials. Appl Environ Microbiol,2009,75:7125~7134.
3 Barton M D. Antibiotic use in animal feed and its impact on human health. Nutr Res Rev,2000,13:279~299.
4 Blake D P,Hillman K,Fenlon D R,et al. Transfer of antibiotic resistance between commensal and pathogenic members of the Enterobacteriaceae under ileal conditions. Journal of Applied Microbiology,2003,95:428~436.
5 Chen J,Yu Z,Michel F C,et al. Development and application of real-time PCR assays for quantification of erm genes conferring resistance to macrocodes-lincosamides-streptogramin B in livestock manure and manure management systems. Appl Environ Microbiol, 2007,73(14):4407~4416.
6 Ghosh S, LaPara T M. The effects of subtherapeutic antibiotic use in farm animals on the proliferation and persistence of antibiotic resistance among soil bacteria.The ISME Journal, 2007, 1:191~203.
7 Lay C,Sutren M,Rochet V,et al. Design and validation of 16S rRNA probes to enumerate members of the Clostridium leptum subgroup in human faecal microbiota. Environ Microbiol,2005,7:933~946.
8 Patterson A J,Colangeli R,Spigaglia P,et al. Distribution of specific tetracycline and erythromycin resistance genes in environmental samples assessed by macroarray detection. Environ Microbiol,2007,9:703~715.
9 Peak N,Knapp C W,Yang R K,et al. Abundance of six tetracycline resistance genes in wastewater lagoons at cattle feedlots with different antibiotic use strategies. Environ Microbiol Letters,2007,9:143~151.
10 Roberts M C. Update on acquired tetracycline resistance genes. FEMS Microbiol Lett,2005,245:195~203.
11 Schmitt H,Stoob K,Hamscher G,et al. Tetracyclines and tetracycline resistance in agricultural soils: Microcosm and field studies. Microbiol Ecol, 2006, 51:267~276.
12 Schuster C J,Ellis A G,Robertson W J,et al. Infectious disease outbreaks related to drinking water in Canada, 1974~2001. Can J Pub Health,Revue Canadienne De Sant Publique, 2005,96(4):254~258.
13 Seville L A,Patterson A J,Scott K P,et al. Distribution of tetracycline and erythromycin resistance genes among human oral and fecal metagenomic DNA. Microb Drug Resist,2009,15:159~166.
14 Shery P G,Chery L W, Josee H,et al. Associations between antimicrobial resistance genes in fecal generic Escherichia coli isolates from cow-calf herds in western Canada. Appl Environ Microbiol,2008,74(12): 3658~3666.
15 Sinton L W,Braithwaite R R,Hall C H,et al. Survival of indicator and pathogenic bacteria in bovine feces on pasture. Appl Environ Microbiol,2007,73:7917~7925.
16 Unc A,Goss M J. Transport of bacteria from manure and protection of water resources. Appl Soil Ecol,2004,25:1~18.

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