Document Detail

Susceptibility of Escherichia coli isolated from uteri of postpartum dairy cows to antibiotic and environmental bacteriophages. Part I: Isolation and lytic activity estimation of bacteriophages.
MedLine Citation:
PMID:  20059908     Owner:  NLM     Status:  MEDLINE    
The objective of this study was to isolate bacteriophages from environmental samples of 2 large commercial dairy farms using Escherichia coli isolated from the uteri of postpartum Holstein dairy cows as hosts. A total of 11 bacteriophage preparations were isolated from manure systems of commercial dairy farms and characterized for in vitro antimicrobial activity. In addition, a total of 57 E. coli uterine isolates from 5 dairy cows were phylogenetically grouped by triplex PCR. Each E. coli bacterial host from the uterus was inoculated with their respective bacteriophage preparation at several different multiplicities of infections (MOI) to determine minimum inhibitory MOI. The effect of a single dose (MOI=10(2)) of bacteriophage on the growth curve of all 57 E. coli isolates was assessed using a microplate technique. Furthermore, genetic diversity within and between the different bacteriophage preparations was assessed by bacteriophage purification followed by DNA extraction, restriction, and agarose gel electrophoresis. Phylogenetic grouping based on triplex PCR showed that all isolates of E. coli belonged to phylogroup B1. Bacterial growth was completely inhibited at considerably low MOI, and the effect of a single dose (MOI=10(2)) of bacteriophage preparations on the growth curve of all 57 E. coli isolates showed that all bacteriophage preparations significantly decreased the growth rate of the isolates. Bacteriophage preparation 1230-10 had the greatest antimicrobial activity and completely inhibited the growth of 71.7% (n=57) of the isolates. The combined action of bacteriophage preparations 1230-10, 6375-10, 2540-4, and 6547-2, each at MOI=10(2), had the broadest spectrum of action and completely inhibited the growth (final optical density at 600 nm <or=0.1) of 80% of the E. coli isolates and considerably inhibited the growth (final optical density at 600 nm <or=0.2) of 90% of the E. coli isolates. Restriction profile analysis demonstrated that all 4 phage preparations contained bacteriophages that were genetically distinct from each other according to the banding pattern of the fragments. The combination of several different bacteriophages can improve the spectrum of action, and the results of this study suggest that bacteriophages 1230-10, 6375-10, 2540-4, and 6547-2 should be used in combination as a cocktail.
R C Bicalho; T M A Santos; R O Gilbert; L S Caixeta; L M Teixeira; M L S Bicalho; V S Machado
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Journal of dairy science     Volume:  93     ISSN:  1525-3198     ISO Abbreviation:  J. Dairy Sci.     Publication Date:  2010 Jan 
Date Detail:
Created Date:  2010-01-11     Completed Date:  2010-04-19     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  2985126R     Medline TA:  J Dairy Sci     Country:  United States    
Other Details:
Languages:  eng     Pagination:  93-104     Citation Subset:  IM    
Copyright Information:
Copyright 2010 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.
Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853, USA.
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MeSH Terms
Bacteriophages / classification,  physiology*
Cattle Diseases / microbiology*
Environmental Microbiology
Escherichia coli / classification,  growth & development,  isolation & purification,  virology*
Escherichia coli Infections / microbiology*
Genetic Variation
Manure / virology
Postpartum Period
Uterus / microbiology*
Reg. No./Substance:

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