Pathogenicity of Enterohemorrhagic Escherichia coli in Neonatal Calves and Evaluation of Fecal Shedding by Treatment with Probiotic Escherichia coli

2003 ◽  
Vol 66 (6) ◽  
pp. 924-930 ◽  
Author(s):  
TONG ZHAO ◽  
SUZANA TKALCIC ◽  
MICHAEL P. DOYLE ◽  
BARRY G. HARMON ◽  
CATHY A. BROWN ◽  
...  

The pathogenicity and fecal shedding of enterohemorrhagic Escherichia coli (EHEC) O26:H11, O111:NM, and O157:H7 were compared in calves (<1 week of age) with or without prior treatment with probiotic bacteria (competitive exclusion E. coli). Three groups of 12 to 14 calves were used for these treatments. Half of the calves in each group were perorally administered 1010 CFU of probiotic bacteria per calf, and, 2 days thereafter, 108 CFU of a five-strain mixture with one of the three EHEC serotypes per calf were administered to each calf. None of the EHEC serotypes caused clinical disease, and neither gross nor microscopic lesions attributable to EHEC were detected in control or probiotic-treated calves at necropsy. In calves administered E. coli O157:H7, fecal shedding was greatly reduced (>6 log10 CFU/g) by 8 days after administration, and there was no significant difference (P > 0.05) in fecal shedding of E. coli O157:H7 between probiotic-treated and untreated control groups at that time. In contrast, control calves perorally administered E. coli of serotypes O111:NM or O26:H11 continued to shed substantial populations (102.1 to 106 CFU/g of feces and 102.5 to 104.9 CFU/g of feces, respectively) throughout 7 days postadministrationof EHEC. In both groups administered either E. coli O111:NM or O26:H11, significantly less (P < 0.05) EHEC was isolated from feces at 7 days postadministration of EHEC and at necropsy from the probiotic-treated group than from the untreated control group. Overall, neonatal calves shed in the feces from 1 to 7 days following peroral administration of EHEC greater populations of E. coli O111:NM and O26:H11 than E. coli O157:H7. In addition, treatment of calves with probiotic E. coli reduced fecal shedding of E. coli O111:NM and O26:H11 in most calves.

2003 ◽  
Vol 66 (7) ◽  
pp. 1184-1189 ◽  
Author(s):  
SUZANA TKALCIC ◽  
TONG ZHAO ◽  
BARRY G. HARMON ◽  
MICHAEL P. DOYLE ◽  
CATHY A. BROWN ◽  
...  

The fecal shedding and pathogenicity of enterohemorrhagic E. coli (EHEC) O26:H11, EHEC O111:NM, and EHEC O157:H7 in weaned calves (8 to 10 weeks of age) were compared with and without treatment with a three-strain mixture of probiotic bacteria (competitive-exclusion E. coli). Three groups of 12 calves were each perorally given a five-strain mixture of one of the EHEC serotypes (1010 CFU of total bacteria per calf). Seventy-two hours later, six calves from each group were each administered 1010 CFU of probiotic bacteria. None of the EHEC serotypes caused significant clinical disease, although a few calves developed mild transient diarrhea or pyrexia. Gross or microscopic lesions attributable to EHEC were not detected in control or probiotic-treated calves at necropsy. For probiotic-treated calves given E. coli O157:H7 and for probiotic-treated calves given E. coli O111:NM, fecal shedding was reduced compared with that for untreated calves. For the probiotic-treated calves given E. coli O157:H7, the reductions in fecal shedding on days 8, 12, 14, 16, 20, 22, 28, and 30 after peroral administration were statistically significant (P < 0.05). For probiotic-treated calves given E. coli O111:NM, there were statistically significant reductions (P < 0.05) in fecal shedding on days 6, 8, 10, and 12. In contrast, there was no reduction in fecal shedding for calves administered E. coli O26:H11 and treated with the probiotic bacteria. In fact, calves in both the treated and the nontreated groups continued to shed large populations of E. coli O26:H11 throughout the 32-day trial. At necropsy, E. coli O157:H7 was isolated from five of six untreated calves and from only two of six probiotic-treated calves. E. coli O111:NM was isolated from four of six untreated calves at necropsy and from two of six probiotic-treated calves. However, E. coli O26:H11 was isolated from five of six untreated calves and from all six probiotic-treated calves. The results obtained in this study indicate that probiotic E. coli substantially reduced or eliminated fecal shedding of E. coli O157:H7 and E. coli O111:NM 8 to 30 days and 6 to 12 days after the administration of the probiotic culture, respectively, and reduced the persistence of E. coli O157:H7 in the gastrointestinal tract at necropsy (31 to 33 days after the administration of the probiotic culture). The probiotic E. coli did not reduce fecal shedding or gastrointestinal persistence of E. coli O26:H11.


2004 ◽  
Vol 70 (10) ◽  
pp. 6053-6060 ◽  
Author(s):  
Gerry P. Schamberger ◽  
Ronald L. Phillips ◽  
Jennifer L. Jacobs ◽  
Francisco Diez-Gonzalez

ABSTRACT A cattle trial using artificially inoculated calves was conducted to determine the effect of the addition of colicinogenic Escherichia coli strains capable of producing colicin E7 (a 61-kDa DNase) to feed on the fecal shedding of serotype O157:H7. The experiment was divided into three periods. In period 1, which lasted 24 days, six calves were used as controls, and eight calves received 107 CFU of E. coli (a mixture of eight colicinogenic E. coli strains) per g of feed. Both groups were orally inoculated with nalidixic acid-resistant E. coli O157:H7 strains 7 days after the treatment started. In periods 2 and 3, the treatment and control groups were switched, and the colicinogenic E. coli dose was increased 10-fold. During period 3, which lasted as long as period 1, both groups were reinoculated with E. coli O157:H7. The numbers of E. coli O157:H7 were consistently greater in the control groups during the three periods, but comparisons within each time period determined a statistically significant (P < 0.05) difference only at day 21 of period 1. However, when the daily average counts were compared between the period 1 control group and the period 3 treatment group that included the same six animals, an overall reduction of 1.1 log10 CFU/g was observed, with a maximum decrease of 1.8 log10 CFU/g at day 21 (overall statistical significance, P = 0.001). Serotype O157:H7 was detected in 44% of the treatment group's intestinal tissue samples and in 64% of those from the control group (P < 0.04). These results indicated that the daily addition of 108 CFU of colicin E7-producing E. coli per gram of feed could reduce the fecal shedding of serotype O157:H7.


1998 ◽  
Vol 36 (3) ◽  
pp. 641-647 ◽  
Author(s):  
Tong Zhao ◽  
Michael P. Doyle ◽  
Barry G. Harmon ◽  
Cathy A. Brown ◽  
P. O. Eric Mueller ◽  
...  

Bacteria inhibitory to Escherichia coli O157:H7 were isolated from cattle and evaluated for their potential for reducing carriage of E. coli O157:H7 in calves. Eighteen of 1,200 bacterial isolates from cattle feces and intestinal tissue samples were screened and determined to inhibit the growth of E. coliO157:H7 in vitro. Seventeen of the isolates were E. coli and one was Proteus mirabilis. None produced Shiga toxin. Genomic DNA fingerprinting by pulsed-field gel electrophoresis revealed 13 distinguishable profiles among the 18 isolates. Two calves inoculated perorally with a mixture of all 18 isolates (1010 CFU) appeared to be normal and did not develop signs of clinical disease throughout a 25- to 27-day observation period. These bacteria colonized segments of the gastrointestinal tract and were in feces at the termination of the experiment (25 and 27 days postinoculation) at levels of 50 to 200 CFU/g. Fifteen cannulated calves were studied to determine the efficiency of the probiotic bacteria in reducing or eliminating the carriage of E. coli O157:H7. Nine calves served as controls, with each animal receiving perorally 1010 CFU ofE. coli O157:H7. E. coliO157:H7 was detected intermittently in the rumen samples from all control animals throughout 3 weeks postinoculation, whereasE. coli O157:H7 was shed at various levels in feces continuously throughout the experiment (mean, 28 days).E. coli O157:H7 was isolated from the rumens and colons of eight of nine and nine of nine calves, respectively, at the termination of the study. Six calves each received perorally 1010 CFU of probiotic bacteria and then 2 days later received 1010 CFU of E. coli O157:H7.E. coli O157:H7 was detected in the rumen for only 9 days postinoculation in two animals, for 16 days in one animal, for 17 days in two animals, and for 29 days in one animal. E. coli O157:H7 was detected in feces for only 11 days postinoculation in one animal, for 15 days in one animal, for 17 days in one animal, for 18 days in one animal, for 19 days in one animal, and for 29 days in one animal. At the end of the experiment (mean, 30 days), E. coli O157:H7 was not recovered from the rumen of any of the six animals treated with probiotic bacteria; however, E. coli O157:H7 was recovered from the feces of one of the animals. This animal was fasted twice postinoculation. These studies indicate that selected probiotic bacteria administered to cattle prior to exposure to E. coli O157:H7 can reduce the level of carriage ofE. coli O157:H7 in most animals.


2008 ◽  
Vol 71 (3) ◽  
pp. 539-544 ◽  
Author(s):  
EBOT S. TABE ◽  
JAMES OLOYA ◽  
DAWN K. DOETKOTT ◽  
MARC L. BAUER ◽  
PENELOPE S. GIBBS ◽  
...  

The effect of direct-fed microbials (DFM) on fecal shedding of Escherichia coli O157:H7 and Salmonella in naturally infected feedlot cattle was evaluated in a clinical trial involving 138 feedlot steers. Following standard laboratory methods, fecal samples collected from steers were evaluated for change in the detectable levels of E. coli O157:H7 and Salmonella shed in feces after DFM treatment. Sampling of steers was carried out every 3 weeks for 84 days. A significant reduction (32%) in fecal shedding of E. coli O157:H7 (P &lt; 0.001), but not Salmonella (P = 0.24), was observed among the treatment steers compared with the control group during finishing. The probability of recovery of E. coli O157:H7 from the feces of treated and control steers was 34.0 and 66.0%, respectively. Steers placed on DFM supplement were almost three times less likely to shed E. coli O157:H7 (odds ratio, 0.36; 95% confidence interval, 0.25 to 0.53; P &lt; 0.001) in their feces as opposed to their control counterparts. The probability of recovery of Salmonella from the feces of the control (14.0%) and the treated (11.3%) steers was similar. However, the DFM significantly reduced probability of new infections with Salmonella among DFM-treated cattle compared with controls (nontreated ones). It appears that DFM as applied in our study are capable of significantly reducing fecal shedding of E. coli O157:H7 in naturally infected cattle but not Salmonella. The factors responsible for the observed difference in the effects of DFM on E. coli O157:H7 and Salmonella warrants further investigation.


2020 ◽  
Vol 8 (2) ◽  
pp. 263 ◽  
Author(s):  
Rodney A. Moxley ◽  
Tom W. Bargar ◽  
Stephen D. Kachman ◽  
Diane R. Baker ◽  
David H. Francis

Enterohemorrhagic Escherichia coli (EHEC), a pathogenic subset of Shiga toxin-producing E. coli (STEC), is an important cause of hemorrhagic colitis and hemolytic–uremic syndrome (HUS), and a rare cause of urinary tract infections (UTIs) with associated HUS. EHEC strains attach intimately to intestinal epithelium with formation of actin pedestals (attaching-effacing (A/E) lesions); however, the mechanism of EHEC attachment to the uroepithelium is unknown. We conducted a retrospective study on archived urinary bladder specimens from gnotobiotic piglets that naturally developed cystitis associated with EHEC O157:H7 infection following oral inoculation and fecal shedding. Paraffin-embedded bladder tissues from three piglets with cystitis and immunohistochemical evidence of EHEC O157:H7 adherence to the uroepithelium were processed for and examined by transmission electron microscopy. EHEC O157:H7 bacteria were found in one of three piglets, intimately attached to pedestals on the apical surfaces of the superficial urothelium (umbrella cells). Cystitis was significantly associated with the length of survival of the piglets post-inoculation (p = 0.0339; estimated odds ratio = 2.6652). This is the first report of E. coli causing A/E-like lesions in the uroepithelium, and also evidence of the utility of the gnotobiotic piglet as a model for studies of the pathogenesis of EHEC UTIs.


2005 ◽  
Vol 71 (1) ◽  
pp. 93-97 ◽  
Author(s):  
J. Christopher Low ◽  
Iain J. McKendrick ◽  
Caroline McKechnie ◽  
David Fenlon ◽  
Stuart W. Naylor ◽  
...  

ABSTRACT Escherichia coli O157:H7 is an important cause of diarrhea, hemorrhagic colitis, and potentially fatal human illness. Cattle are considered a primary reservoir of infection, and recent experimental evidence has indicated that the terminal rectum is the principal site of bacterial carriage. To test this finding in naturally colonized animals, intact rectum samples from 267 cattle in 24 separate lots were obtained immediately after slaughter, and fecal material and mucosal surfaces were cultured for E. coli O157 by direct and enrichment methods. Two locations, 1 and 15 cm proximal to the recto-anal junction, were tested. In total, 35 animals were positive for E. coli O157 at at least one of the sites and 232 animals were negative as determined by all tests. The frequency of isolation and the numbers of E. coli O157 cells were higher at the site closer to the recto-anal junction, confirming our previous experimental findings. We defined low- and high-level carriers as animals with E. coli O157 levels of <1 � 103 CFU g−1 or <1 � 103 CFU ml−1 and animals with E. coli O157 levels of ≥1 � 103 CFU g−1 or ≥1 � 103 CFU ml−1 in feces or tissues, respectively. High-level carriage was detected in 3.7% of the animals (95% confidence interval, 1.8 to 6.8%), and carriage on the mucosal surface of the terminal rectum was associated with high-level fecal excretion. In summary, our results support previous work demonstrating that the mucosal epithelium in the bovine terminal rectum is an important site for E. coli O157 carriage in cattle. The data also support the hypothesis that high-level fecal shedding (≥1 � 103 CFU g of feces−1) of enterohemorrhagic E. coli O157 results from colonization of this site.


1999 ◽  
Vol 62 (7) ◽  
pp. 747-750 ◽  
Author(s):  
KUMAR S. VENKITANARAYANAN ◽  
TONG ZHAO ◽  
MICHAEL P. DOYLE

The antibacterial activity of lactoferricin B on enterohemorrhagic Escherichia coli O157:H7 in 1% peptone medium and ground beef was studied at 4 and 10°C. In 1% peptone medium, 50 and 100 μg of lactoferricin B per ml reduced E. coli O157:H7 populations by approximately 0.7 and 2.0 log CFU/ml, respectively. Studies comparing the antibacterial effect of lactoferricin B on E. coli O157:H7 in 1% peptone at pH 5.5 and 7.2 did not reveal any significant difference (P &gt; 0.5) at the two pH values. Lactoferricin B (100 μg/g) reduced E. coli O157:H7 population in ground beef by about 0.8 log CFU/g (P &lt; 0.05). No significant difference (P &gt; 0.5) was observed in the total plate count between treatment and control ground beef samples stored at 4 and 10°C. The antibacterial effect of lactoferricin B on E. coli O157:H7 observed in this study is not of sufficient magnitude to merit its use in ground beef for controlling the pathogen.


2010 ◽  
Vol 76 (21) ◽  
pp. 7210-7216 ◽  
Author(s):  
Lucia Rivas ◽  
Brid Coffey ◽  
Olivia McAuliffe ◽  
Mary J. McDonnell ◽  
Catherine M. Burgess ◽  
...  

ABSTRACT This study investigated the effect of bacteriophages (phages) e11/2 and e4/1c against Escherichia coli O157:H7 in an ex vivo rumen model and in cattle in vivo. In the ex vivo rumen model, samples were inoculated with either 103 or 106 CFU/ml inoculum of E. coli O157:H7 and challenged separately with each bacteriophage. In the presence of phage e11/2, the numbers of E. coli O157:H7 bacteria were significantly (P < 0.05) reduced to below the limit of detection within 1 h. Phage e4/1c significantly (P < 0.05) reduced E. coli O157:H7 numbers within 2 h of incubation, but the number of surviving E. coli O157:H7 bacteria then remained unchanged over a further 22-h incubation period. The ability of a phage cocktail of e11/2 and e4/1c to reduce the fecal shedding of E. coli O157:H7 in experimentally inoculated cattle was then investigated in two cattle trials. Cattle (yearlings, n = 20 for trial one; adult fistulated cattle, n = 2 for trial two) were orally inoculated with 1010 CFU of E. coli O157:H7. Animals (n = 10 for trial one; n = 1 for trial two) were dosed daily with a bacteriophage cocktail of 1011 PFU for 3 days postinoculation. E. coli O157:H7 and phage numbers in fecal and/or rumen samples were determined over 7 days postinoculation. E. coli O157:H7 numbers rapidly declined in all animals within 24 to 48 h; however, there was no significant difference (P > 0.05) between the numbers of E. coli O157:H7 bacteria shed by the phage-treated or control animals. Phages were recovered from the rumen but not from the feces of the adult fistulated animal in trial two but were recovered from the feces of the yearling animals in trial one. While the results from the rumen model suggest that phages are effective in the rumen, further research is required to improve the antimicrobial effectiveness of phages for the elimination of E. coli O157:H7 in vivo.


2000 ◽  
Vol 63 (11) ◽  
pp. 1467-1474 ◽  
Author(s):  
S. J. BUCHKO ◽  
R. A. HOLLEY ◽  
W. O. OLSON ◽  
V. P. J. GANNON ◽  
D. M. VEIRA

Three groups of six yearling steers (three rumen fistulated plus three nonfistulated) fed one of three different grain diets (85% cracked corn, 15% whole cottonseed and 70% barley, or 85% barley) were inoculated with 1010 CFU of Escherichia coli O157:H7 strain 3081, and the presence of the inoculated strain was followed in the rumen fluid and feces for a 10-week period. E. coli O157:H7 was rapidly eliminated from the rumen of the animals on all three diets but persisted in the feces of some animals up to 67 days after inoculation, suggesting that the bovine hindgut is the site of E. coli O157:H7 persistence. A significant difference existed in the levels of E. coli O157:H7 shed by the animals among diets on days 5, 7, 49, and 63 after inoculation (P &lt; 0.05). No significant difference was found between the levels shed among diets on days 9 through 42 and on day 67 (P &gt; 0.05). The number of animals that were culture positive for E. coli O157:H7 strain 3081 during the 10-week period was significantly higher for the barley fed group (72 of 114 samplings) as opposed to the corn fed group (44 of 114 samplings) (P &lt; 0.005) and the cottonseed and barley fed group (57 of 114 samplings) (P &lt; 0.05). The fecal pH of the animals fed the corn diet was significantly lower (P &lt; 0.05) than the fecal pH of the animals fed the cottonseed and barley and barley diets, likely resulting in a less suitable environment for E. coli O157:H7 in the hindgut of the corn fed animals. E. coli O157:H7 strain 3081 was present in 3 of 30 (corn, 1 of 10; cottonseed, 1 of 10; barley, 1 of 10) animal drinking water samples, 3 of 30 (corn, 1 of 10; cottonseed, 0 of 10; barley, 2 of 10) water trough biofilm swabs, 5 of 30 (corn, 0 of 10; cottonseed, 2 of 10; barley, 3 of 10) feed samples, and 30 of 30 manure samples taken from the pens during the entire experimental period. Mouth swabs of the steers were also culture positive for E. coli O157:H7 strain 3081 in 30 of 180 samples (corn, 7 of 60; cottonseed, 4 of 60; barley, 19 of 60) taken during the 10-week period. Minimizing environmental dissemination of E. coli O157:H7 in conjunction with diet modification may reduce numbers of E. coli O157:H7–positive cattle.


2009 ◽  
Vol 89 (2) ◽  
pp. 285-293 ◽  
Author(s):  
S J Bach ◽  
R P Johnson ◽  
K. Stanford ◽  
T A McAllister

Bacteriophage biocontrol has potential as a means of mitigating the prevalence of Escherichia coli O157:H7 in ruminants. The efficacy of oral administration of bacteriophages for reducing fecal shedding of E. coli O157:H7 by sheep was evaluated using 20 Canadian Arcott rams (50.0 ± 3.0) housed in four rooms (n = 5) in a contained facility. The rams had ad libitum access to drinking water and a pelleted barley-based total mixed ration, delivered once daily. Experimental treatments consisted of administration of E. coli O157:H7 (O157), E. coli O157:H7+bacteriophages (O157+phage), bacteriophages (phage), and control (CON). Oral inoculation of the rams with 109 CFU of a mixture of four nalidixic acid-resistant strains of E. coli O157:H7 was performed on day 0. A mixture of 1010 PFU of bacteriophages P5, P8 and P11 was administered on days -2, -1, 0, 6 and 7. Fecal samples collected on 14 occasions over 21 d were analyzed for E. coli O157:H7, total E. coli, total coliforms and bacteriophages. Sheep in treatment O157+phage shed fewer (P < 0.05) E. coli O157:H7 than did sheep in treatment O157. Populations of total coliforms and total E. coli were similar (P < 0.05) among treatments, implying that bacteriophage lysis of non-target E. coli and coliform bacteria in the gastrointestinal tract did not occur. Bacteriophage numbers declined rapidly over 21 d, which likely reduced the chance of collision between bacteria and bacteriophage. Oral administration of bacteriophages reduced shedding of E. coli O157:H7 by sheep, but a delivery system that would protect bacteriophages during passage through the intestine may increase the effectiveness of this strategy as well as allow phage to be administered in the feed.Key words: Escherichia coli O157:H7, bacteriophage, sheep, environment, coliforms


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