scholarly journals Quantitative PCR Profiling of Escherichia coli in Livestock Feces Reveals Increased Population Resilience Relative to Culturable Counts under Temperature Extremes

2016 ◽  
Vol 50 (17) ◽  
pp. 9497-9505 ◽  
Author(s):  
David M. Oliver ◽  
Clare Bird ◽  
Emmy Burd ◽  
Michael Wyman
Keyword(s):  
Escherichia Coli ◽  
Quantitative Pcr ◽  
Temperature Extremes ◽  
Population Resilience ◽  
Livestock Feces

Multiplex Quantitative PCR Assays for the Detection and Quantification of the Six Major Non-O157 Escherichia coli Serogroups in Cattle Feces†

2016 ◽  
Vol 79 (1) ◽  
pp. 66-74 ◽  
Author(s):  
P. B. SHRIDHAR ◽  
L. W. NOLL ◽  
X. SHI ◽  
B. AN ◽  
N. CERNICCHIARO ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Quantitative Pcr ◽  
Foodborne Pathogens ◽  
Virulence Genes ◽  
Target Genes ◽  
Culture Methods ◽  
Fecal Samples ◽  
E Coli ◽  
Cattle Feces ◽  
Before And After

ABSTRACT Shiga toxin–producing Escherichia coli (STEC) serogroups O26, O45, O103, O111, O121, and O145, called non-O157 STEC, are important foodborne pathogens. Cattle, a major reservoir, harbor the organisms in the hindgut and shed them in the feces. Although limited data exist on fecal shedding, concentrations of non-O157 STEC in feces have not been reported. The objectives of our study were (i) to develop and validate two multiplex quantitative PCR (mqPCR) assays, targeting O-antigen genes of O26, O103, and O111 (mqPCR-1) and O45, O121, and O145 (mqPCR-2); (ii) to utilize the two assays, together with a previously developed four-plex qPCR assay (mqPCR-3) targeting the O157 antigen and three virulence genes (stx1, stx2, and eae), to quantify seven serogroups and three virulence genes in cattle feces; and (iii) to compare the three mqPCR assays to a 10-plex conventional PCR (cPCR) targeting seven serogroups and three virulence genes and culture methods to detect seven E. coli serogroups in cattle feces. The two mqPCR assays (1 and 2) were shown to be specific to the target genes, and the detection limits were 4 and 2 log CFU/g of pure culture–spiked fecal samples, before and after enrichment, respectively. A total of 576 fecal samples collected from a feedlot were enriched in E. coli broth and were subjected to quantification (before enrichment) and detection (after enrichment). Of the 576 fecal samples subjected, before enrichment, to three mqPCR assays for quantification, 175 (30.4%) were quantifiable (≥4 log CFU/g) for at least one of the seven serogroups, with O157 being the most common serogroup. The three mqPCR assays detected higher proportions of postenriched fecal samples (P < 0.01) as positive for one or more serogroups compared with cPCR and culture methods. This is the first study to assess the applicability of qPCR assays to detect and quantify six non-O157 serogroups in cattle feces and to generate data on fecal concentration of the six serogroups.

scholarly journals Rapid Onset of Ulcerative Typhlocolitis in B6.129P2-IL10tm1Cgn (IL-10−/−) Mice Infected with Helicobacter trogontum Is Associated with Decreased Colonization by Altered Schaedler's Flora

2006 ◽  
Vol 74 (12) ◽  
pp. 6615-6623 ◽  
Author(s):  
M. T. Whary ◽  
S. J. Danon ◽  
Y. Feng ◽  
Z. Ge ◽  
N. Sundina ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Quantitative Pcr ◽  
Body Condition ◽  
Experimental Infection ◽  
Clinical Signs ◽  
Interleukin 10 ◽  
Rapid Onset ◽  
Lower Bowel ◽  
Severe Diarrhea ◽  
First Time

ABSTRACT Infection with Helicobacter trogontum, a urease-positive helicobacter isolated from subclinically infected rats, was evaluated in B6.129P2-IL10 tm1Cgn (interleukin-10−/− [IL-10−/−]) and C57BL/6 (B6) mice. In a first experiment, IL-10−/− mice naturally infected with Helicobacter rodentium had subclinical typhlocolitis but developed severe diarrhea and loss of body condition with erosive to ulcerative typhlocolitis within 1 to 3 weeks of experimental infection with H. trogontum. A second experiment demonstrated that helicobacter-free IL-10−/− mice dosed with H. trogontum also developed severe clinical signs and typhlocolitis within 2 to 4 weeks, whereas B6 mice colonized with H. trogontum were resistant to disease. In a third experiment, using helicobacter-free IL-10−/− mice, dosing with H. trogontum resulted in acute morbidity and typhlocolitis within 8 days. Acute typhlocolitis was accompanied by signs of sepsis supported by degenerative hemograms and recovery of Escherichia coli and Proteus spp. from the livers of infected mice. Quantitative PCR data revealed that H. rodentium and H. trogontum may compete for colonization of the lower bowel, as H. trogontum established higher colonization levels in the absence of H. rodentium (P < 0.003). H. trogontum-induced typhlocolitis was also associated with a significant decrease in the levels of colonization by five of eight anaerobes that comprise altered Schaedler's flora (P < 0.002). These results demonstrate for the first time that H. rodentium infection in IL-10−/− mice causes subclinical typhlocolitis and that infection with H. trogontum (with or without H. rodentium) induces a rapid-onset, erosive to ulcerative typhlocolitis which impacts the normal anaerobic flora of the colon and increases the risk of sepsis.

scholarly journals Comparison of quantitative PCR assays for Escherichia coli targeting ribosomal RNA and single copy genes

2011 ◽  
Vol 52 (3) ◽  
pp. 298-306 ◽  
Author(s):  
E.C. Chern ◽  
S. Siefring ◽  
J. Paar ◽  
M. Doolittle ◽  
R.A. Haugland
Keyword(s):  
Escherichia Coli ◽  
Quantitative Pcr ◽  
Ribosomal Rna ◽  
Single Copy ◽  
Pcr Assays

scholarly journals Diversity and Abundance of Zoonotic Pathogens and Indicators in Manures of Feedlot Cattle in Australia

2010 ◽  
Vol 76 (20) ◽  
pp. 6947-6950 ◽  
Author(s):  
Marcus Klein ◽  
Leearna Brown ◽  
Robyn W. Tucker ◽  
Nicholas J. Ashbolt ◽  
Richard M. Stuetz ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Quantitative Pcr ◽  
Campylobacter Jejuni ◽  
Dry Weight ◽  
Feedlot Cattle ◽  
Fecal Indicators ◽  
Zoonotic Pathogens

ABSTRACT The occurrence of 10 pathogens and three fecal indicators was assessed by quantitative PCR in manures of Australian feedlot cattle. Most samples tested positive for one or more pathogens. For the dominant pathogens Campylobacter jejuni, Listeria monocytogenes, Giardia spp., Cryptosporidium spp., and eaeA-positive Escherichia coli, 102 to 107 genome copies g−1 (dry weight) manure were recovered.

Real time quantitative PCR of Bacteroides vulgatus and Escherichia coli in mucosal samples from healthy humans and patients with IBD

2000 ◽  
Vol 118 (4) ◽  
pp. A1347
Author(s):  
Xander W. Huijsdens ◽  
Ronald K. Linskens ◽  
Mariette Mak ◽  
Jeroen Stoof ◽  
Paul Hm Savelkoul ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Real Time ◽  
Quantitative Pcr ◽  
Real Time Quantitative Pcr ◽  
Healthy Humans
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