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

doi:10.1128/JCM.36.12.3480-3487.1998

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

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Table 1.

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

Farm	Date of sampling	No. of cattle O157 VTEC positive/total no. of cattle (%)	Phage type	Result of Vero cell assay for VT	PCR results for the following:			PFGE pattern	No. of isolates
Farm	Date of sampling	No. of cattle O157 VTEC positive/total no. of cattle (%)	Phage type	Result of Vero cell assay for VT	VT1	VT2	eae	PFGE pattern	No. of isolates
I	Sept 1996	27/140 (19.3)	34	+	+	+	+	A	23
			34	+	−	+	+	A	2
			31	+	−	+	+	B	1
			Rdnc^a	+	+	+	+	C	1
II	Sept 1996	15/67 (22.4)	8	+	+	+	+	D	15
III	Sept 1996	7/112 (6.3)	31	+	−	+	+	E	7
IV	Oct 1996	3/162 (1.9)	Rdnc	+	−	+	+	F	3
V	Oct 1996	17/83 (20.5)	14	+	−	+	+	G	17
VI	Oct 1996	1/120 (0.8)	14	+	−	+	+	H	1
VII	Nov 1996	5/100 (5.0)	8	+	+	+	+	I	5
VIII	Nov 1996	0/100 (0.0)
IX	Nov 1996	0/195 (0.0)
X	Nov 1996	0/73 (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

Farm	Date of sampling	No. of cattle O157 VTEC positive/total no. of cattle (%)	Phage type	Result of Vero cell assay for VT	PCR results for the following:			PFGE pattern	No. of isolates
Farm	Date of sampling	No. of cattle O157 VTEC positive/total no. of cattle (%)	Phage type	Result of Vero cell assay for VT	VT1	VT2	eae	PFGE pattern	No. of isolates
II	Sept 1996	15/67 (22.4)	8	+	+	+	+	D	15
	Dec 1996	8/106 (7.5)	8	+	+	+	+	J	7
			8	+	+	+	+	D	1^a
	Mar 1997	0/99 (0.0)
	Jun 1997	0/99 (0.0)
	Sept 1997	36/59 (61.0)	21	+	−	+	+	K	35^b
			54	+	−	+	+	L	1
	Nov 1997	15/104 (14.4)	21	+	−	+	+	K	15^c
V	Oct 1996	17/83 (20.5)	14	+	−	+	+	G	17
	Jan 1997	1/83 (1.2)	14	+	−	+	+	G	1^d
	Apr 1997	2/79 (2.5)	4	+	−	+	+	M	2^e
	Jun 1997	9/52 (17.3)	14	+	−	+	+	N	8^f
			48^g	+	−	+	+	N	1
	Sept 1997	2/38 (5.3)	14	+	−	+	+	O	1^h
			54	+	−	+	+	P	1
	Nov 1997	0/41 (0.0)
VIII	Nov 1996	0/100 (0.0)
	Jan 1997	0/129 (0.0)
	Apr 1997	1/131 (0.8)	4	+	−	+	+	M	1
	Jul 1997	0/99 (0.0)
	Sept 1997	0/81 (0.0)
	Nov 1997	0/110 (0.0)
X	Nov 1996	0/73 (0.0)
	Feb 1997	0/73 (0.0)
	Apr 1997	0/64 (0.0)
	Sept 1997	2/22 (9.1)	34	+	−	+	+	Q	2
	Nov 1997	1/74 (1.4)	34	+	−	+	+	Q	1

↵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

Farm	Date of sampling	No. of samples O157 VTEC positive/no. of samples tested
Farm	Date of sampling	Cattle^a	Pigs	Horses	Ponies	Sheep	Stable flies	Milk filter	Silage	Animal drinking water
II	Sept 1996			1^b/2^c	0/1
	Dec 1996				1^d/1
	Mar 1997	0/1		0/4	0/1			0/1		0/1
	Jun 1997			0/1	0/2			0/1
	Sept 1997	0/1		3^e/3^c	1^e/1		1^e/1	1^e/1		0/1
	Nov 1997			0/2	0/3		0/1	0/1	0/2	0/2
V	Oct 1996^f		0/10			1^g/1		1^g/1	0/2	0/1
	Jan 1997^h	0/2ⁱ	0/6			4^g/5		0/1	0/3
	Apr 1997	0/3^j	0/3			0/5		0/1
	Jun 1997	1^k,l/5	0/5			1^l/4		0/1
	Sept 1997	0/5^m	0/4				0/1	0/1
	Nov 1997	1ⁿ/7	0/5		0/2	0/9		0/1		0/2
VIII	Nov 1996		0/15					0/1	0/1	0/1
	Jan 1997							0/1		0/1
	Apr 1997	0/1	0/5					0/1		0/1
	Jul 1997		0/5					0/1		0/1
	Sept 1997	0/1	0/3				0/1	0/1		0/1
	Nov 1997		0/5					0/1	0/1	0/1
X	Nov 1996		0/7					0/1	0/1	0/1
	Feb 1997		0/4					0/1	0/1	0/1
	Apr 1997		0/3					0/1		0/1
	Sept 1997	14^o/18	0/4					0/1		0/2
	Nov 1997							0/1	0/1	0/1

↵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.