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Clinical Veterinary Microbiology

Occurrence of Verocytotoxin-ProducingEscherichia coli O157 on Dutch Dairy Farms

A. E. Heuvelink, F. L. A. M. van den Biggelaar, J. T. M. Zwartkruis-Nahuis, R. G. Herbes, R. Huyben, N. Nagelkerke, W. J. G. Melchers, L. A. H. Monnens, E. de Boer
A. E. Heuvelink
Departments of Medical Microbiology and
Pediatrics, University Hospital Nijmegen, 6500 HB Nijmegen,
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F. L. A. M. van den Biggelaar
Inspectorate for Health Protection, Food Inspection Service, 7200 GN Zutphen,
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J. T. M. Zwartkruis-Nahuis
Inspectorate for Health Protection, Food Inspection Service, 7200 GN Zutphen,
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R. G. Herbes
Veterinary Public Health Inspectorate, 6800 DR Arnhem,
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R. Huyben
Animal Health Service, 7400 AA Deventer, and
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N. Nagelkerke
National Institute of Public Health and the Environment, Methodological Consultancy Unit, 3720 BA Bilthoven, The Netherlands
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W. J. G. Melchers
Departments of Medical Microbiology and
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L. A. H. Monnens
Pediatrics, University Hospital Nijmegen, 6500 HB Nijmegen,
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E. de Boer
Inspectorate for Health Protection, Food Inspection Service, 7200 GN Zutphen,
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DOI: 10.1128/JCM.36.12.3480-3487.1998
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    Fig. 1.

    Fecal excretion of O157 VTEC by cattle from the seven farms that tested positive in the prevalence study relative to age of cattle. □, percentage of cattle found to be O157 VTEC positive in the age group; •, estimated percentage of O157 VTEC-positive cattle in the age group by GEE analysis; *, statistically significantly different (P < 0.05). Values above the bars are number of cattle found to be O157 VTEC positive/total number of cattle examined in the age group.

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    Fig. 2.

    Fecal excretion of O157 VTEC by cattle from the four farms examined in the follow-up study relative to season and age of cattle. Heifers ages 1 to 2 years could not be sampled in the period from July to September because they were continuously at pasture. Bars, total percentage of cattle found to be O157 VTEC positive in the quarter; •, estimated total percentage of O157 VTEC-positive cattle in the quarter by GEE analysis; *, statistically significantly different (P < 0.05). Values above the bars are number of cattle found to be O157 VTEC-positive/total number of cattle examined in the quarter.

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    Fig. 3.

    Fecal excretion of O157 VTEC by cattle from the four farms examined in the follow-up study relative to age of cattle. □, percentage of cattle found to be O157 VTEC positive in the age group; •, estimated percentage of O157 VTEC-positive cattle in the age group by GEE analysis; ↔, statistically significant difference (P < 0.05) between estimated percentage of O157 VTEC-positive cattle in the age groups. Values above the bars are number of cattle found to be O157 VTEC positive/total number of cattle examined in the age group.

Tables

  • Figures
  • Table 1.

    Isolation and characterization of fecal O157 VTEC strains from cattle from the 10 Dutch dairy farms investigated in the prevalence study

    FarmDate of samplingNo. of cattle O157 VTEC positive/total no. of cattle (%)Phage typeResult of Vero cell assay for VTPCR results for the following:PFGE patternNo. of isolates
    VT1VT2eae
    ISept 199627/140 (19.3)34++++A23
    34+−++A2
    31+−++B1
    Rdnca++++C1
    IISept 199615/67 (22.4)8++++D15
    IIISept 19967/112 (6.3)31+−++E7
    IVOct 19963/162 (1.9)Rdnc+−++F3
    VOct 199617/83 (20.5)14+−++G17
    VIOct 19961/120 (0.8)14+−++H1
    VIINov 19965/100 (5.0)8++++I5
    VIIINov 19960/100 (0.0)
    IXNov 19960/195 (0.0)
    XNov 19960/73 (0.0)
    • ↵a Rdnc, reacted with the phage set but did not correspond to a recognized phage type.

  • Table 2.

    Isolation and characterization of fecal O157 VTEC strains from cattle from the four Dutch dairy farms investigated in the follow-up study

    FarmDate of samplingNo. of cattle O157 VTEC positive/total no. of cattle (%)Phage typeResult of Vero cell assay for VTPCR results for the following:PFGE patternNo. of isolates
    VT1VT2eae
    IISept 199615/67 (22.4)8++++D15
    Dec 19968/106 (7.5)8++++J7
    8++++D1a
    Mar 19970/99 (0.0)
    Jun 19970/99 (0.0)
    Sept 199736/59 (61.0)21+−++K35b
    54+−++L1
    Nov 199715/104 (14.4)21+−++K15c
    VOct 199617/83 (20.5)14+−++G17
    Jan 19971/83 (1.2)14+−++G1d
    Apr 19972/79 (2.5)4+−++M2e
    Jun 19979/52 (17.3)14+−++N8f
    48g+−++N1
    Sept 19972/38 (5.3)14+−++O1h
    54+−++P1
    Nov 19970/41 (0.0)
    VIIINov 19960/100 (0.0)
    Jan 19970/129 (0.0)
    Apr 19971/131 (0.8)4+−++M1
    Jul 19970/99 (0.0)
    Sept 19970/81 (0.0)
    Nov 19970/110 (0.0)
    XNov 19960/73 (0.0)
    Feb 19970/73 (0.0)
    Apr 19970/64 (0.0)
    Sept 19972/22 (9.1)34+−++Q2
    Nov 19971/74 (1.4)34+−++Q1
    • ↵a Isolate from a calf 4 to 12 months of age which also tested positive in September 1996.

    • ↵b Including two isolates from adult cows >3 years of age which also tested positive in September 1996 and three isolates from cattle ages 2 to 3 years which also tested positive in December 1996.

    • ↵c Including one isolate from a calf younger than age 4 months and three isolates from calves 4 to 12 months of age which all also tested positive in September 1997, one isolate from a heifer 1 to 2 years of age which also tested positive in December 1996, and one isolate from a heifer 1 to 2 years of age which also tested positive in September 1996.

    • ↵d Isolate from a calf 4 to 12 months of age which also tested positive in October 1996.

    • ↵e Including an isolate from an adult cow which also tested positive in October 1996.

    • ↵f Including an isolate from a calf 4 to 12 months of age which also tested positive in October 1996.

    • ↵g Phage type 48 differs from phage type 14 in only one phage reaction.

    • ↵h Isolate from an adult cow which also tested positive in October 1996 and April 1997.

  • Table 3.

    Isolation and characterization of O157 VTEC strains from additional samples collected at the four Dutch dairy farms investigated in the follow-up study

    FarmDate of samplingNo. of samples O157 VTEC positive/no. of samples tested
    CattleaPigsHorsesPoniesSheepStable fliesMilk filterSilageAnimal drinking water
    IISept 19961b/2c0/1
    Dec 19961d/1
    Mar 19970/10/40/10/10/1
    Jun 19970/10/20/1
    Sept 19970/13e/3c1e/11e/11e/10/1
    Nov 19970/20/30/10/10/20/2
    VOct 1996f0/101g/11g/10/20/1
    Jan 1997h0/2i0/64g/50/10/3
    Apr 19970/3j0/30/50/1
    Jun 19971k,l/50/51l/40/1
    Sept 19970/5m0/40/10/1
    Nov 19971n/70/50/20/90/10/2
    VIIINov 19960/150/10/10/1
    Jan 19970/10/1
    Apr 19970/10/50/10/1
    Jul 19970/50/10/1
    Sept 19970/10/30/10/10/1
    Nov 19970/50/10/10/1
    XNov 19960/70/10/10/1
    Feb 19970/40/10/10/1
    Apr 19970/30/10/1
    Sept 199714o/180/40/10/2
    Nov 19970/10/10/1
    • ↵a Fecal samples from cattle which could not be identified individually since either the cattle wore no ear tag numbers or the samples were taken from fresh droppings in farm building (adult steers) or in pastures (heifers).

    • ↵b The isolate was characterized as being phage type 8, Vero cell assay positive, VT1 positive, VT2 positive,eae positive, and PFGE pattern D.

    • ↵c Including one fowl.

    • ↵d The isolate was characterized as being phage type 32, Vero cell assay positive, VT1 positive, VT2 positive,eae positive, and PFGE pattern S.

    • ↵e The isolates were characterized as being phage type 21, Vero cell assay positive, VT1 negative, VT2 positive,eae positive, and PFGE pattern K.

    • ↵f Additionally, fecal samples from a dog, a rabbit, and the pig’s dung were collected. All tested negative.

    • ↵g The isolates were characterized as being phage type 14, Vero cell assay positive, VT1 negative, VT2 positive,eae positive, and PFGE pattern G.

    • ↵h Additionally, fecal samples from a dog and the pig’s dung were collected. Both tested negative.

    • ↵i Including one adult steer.

    • ↵j All adult steers.

    • k An adult steer.

    • ↵l The isolate was characterized as being phage type 14, Vero cell assay positive, VT1 negative, VT2 positive,eae positive, and PFGE pattern N.

    • ↵m Including three steers.

    • ↵n Sample taken from a fresh dropping in the pasture. The isolate was characterized as being phage type 14, Vero cell assay positive, VT1 negative, VT2 positive, eaepositive, and PFGE pattern N.

    • ↵o Samples taken from fresh droppings in the pasture. Twelve of the isolates were characterized as being phage 34, Vero cell assay positive, VT1 negative, VT2 positive, eaepositive, and PFGE pattern Q, and the remaining two were characterized as being phage type 2, Vero cell assay positive, VT1 negative, VT2 positive, eae positive, and PFGE pattern T.

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Occurrence of Verocytotoxin-ProducingEscherichia coli O157 on Dutch Dairy Farms
A. E. Heuvelink, F. L. A. M. van den Biggelaar, J. T. M. Zwartkruis-Nahuis, R. G. Herbes, R. Huyben, N. Nagelkerke, W. J. G. Melchers, L. A. H. Monnens, E. de Boer
Journal of Clinical Microbiology Dec 1998, 36 (12) 3480-3487; DOI: 10.1128/JCM.36.12.3480-3487.1998

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Occurrence of Verocytotoxin-ProducingEscherichia coli O157 on Dutch Dairy Farms
A. E. Heuvelink, F. L. A. M. van den Biggelaar, J. T. M. Zwartkruis-Nahuis, R. G. Herbes, R. Huyben, N. Nagelkerke, W. J. G. Melchers, L. A. H. Monnens, E. de Boer
Journal of Clinical Microbiology Dec 1998, 36 (12) 3480-3487; DOI: 10.1128/JCM.36.12.3480-3487.1998
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KEYWORDS

Bacterial Toxins
cattle
Escherichia coli

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