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Strain Variation within Species in Fecal Samples from Dogs and Cats
Miriam G.J. Koene, Dirk J. Houwers, Jeroen R. Dijkstra, Birgitta Duim, Jaap A. Wagenaar
To cite this version:
Miriam G.J. Koene, Dirk J. Houwers, Jeroen R. Dijkstra, Birgitta Duim, Jaap A. Wagenaar. Strain
Variation within Species in Fecal Samples from Dogs and Cats. Veterinary Microbiology, Elsevier,
2008, 133 (1-2), pp.199. �10.1016/j.vetmic.2008.06.022�. �hal-00532447�
Accepted Manuscript
Title: Strain Variation within Campylobacter Species in Fecal Samples from Dogs and Cats
Authors: Miriam G.J. Koene, Dirk J. Houwers, Jeroen R.
Dijkstra, Birgitta Duim, Jaap A. Wagenaar
PII: S0378-1135(08)00257-5
DOI: doi:10.1016/j.vetmic.2008.06.022
Reference: VETMIC 4088
To appear in: VETMIC Received date: 1-4-2008 Revised date: 16-6-2008 Accepted date: 26-6-2008
Please cite this article as: Koene, M.G.J., Houwers, D.J., Dijkstra, J.R., Duim, B., Wagenaar, J.A., Strain Variation within Campylobacter Species in Fecal Samples from Dogs and Cats, Veterinary Microbiology (2007), doi:10.1016/j.vetmic.2008.06.022 This is a PDF file of an unedited manuscript that has been accepted for publication.
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Accepted Manuscript
Strain Variation within Campylobacter Species in Fecal Samples from Dogs and Cats 1
2
Miriam G.J. Koene a,c,d,1*, Dirk J. Houwers a, Jeroen R. Dijkstra b,2, Birgitta Duim b,3, 3
Jaap A. Wagenaar a,b,c,d 4
5 6
a. Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, 7
Utrecht University, P.O. Box 80165, 3508 TD Utrecht, The Netherlands.
8
b. Central Veterinary Institute of Wageningen UR, P.O. Box 65, 8200 AB, Lelystad, The 9
Netherlands.
10
c. OIE Reference Laboratory for campylobacteriosis, The Netherlands.
11
d. WHO Collaborating Centre for Campylobacter, The Netherlands.
12
1. present address: Central Veterinary Institute of Wageningen UR, P.O. Box 65, 8200 AB, 13
Lelystad, The Netherlands 14
2. present address: Dept. of Pathology, Radboud University Nijmegen Medical Centre, 15
Nijmegen, The Netherlands 16
3. present address: Academic Medical Centre, Dept. Medical Microbiology, Amsterdam, The 17
Netherlands 18
19
*Corresponding author:
20
M.G.J. Koene, 21
Phone: +31 320 238425 22
Fax: +31 320 238961 23
Electronic mail address: miriam.koene@wur.nl 24
Manuscript
Accepted Manuscript
Abstract 1
To investigate the incidence of co-colonization of different strains of Campylobacter species 2
present in canine and feline stool samples, isolates were recovered by culture from 40 samples 3
from dogs (n=34) and cats (n=6). Animals were of different ages, with diarrhoea or without 4
clinical signs. Three isolation procedures were used: two selective agars and a filtration 5
method. In each stool sample, multiple colonies were identified to the species level by PCR, 6
subsequently genotyped by AFLP and pattern similarities (451 isolates) were calculated to 7
investigate their phylogenetic relationships.
8
Genetic heterogeneity of strains in individual stool samples was detected within the species 9
Campylobacter jejuni , C. upsaliensis and C. helveticus, though to a different degree in dogs 10
and cats. In 3 of the 34 (9%) canine samples, more than one genotype of the same 11
Campylobacter species was present, while strain variation was detected in four of the six 12
feline samples. The results show that preferably, multiple colonies should be analysed in 13
molecular epidemiological and aetiological studies.
14 15
Keywords 16
Campylobacter jejuni 17
Campylobacter upsaliensis 18
Campylobacter helveticus 19
Co-colonization 20
AFLP 21
strain variation 22
dogs 23
cats 24
25
Accepted Manuscript
Introduction 1
Most Campylobacter species, like C. jejuni, C. coli, C. lari and C. upsaliensis, show a 2
genetically diverse population. Natural transformability and intragenomic DNA 3
rearrangements contribute to this diversity (Boer et al., 2002). Co-colonization of multiple 4
genotypes of a Campylobacter species has been shown to occur in humans, pigs and poultry 5
(Richardson et al., 2001; Schouls et al., 2003; Weijtens et al., 1999) and is thought to facilitate 6
DNA exchange and therewith increase the genetic diversity.
7
In both dogs and cats, long term colonisation of Campylobacter (Bender et al., 2005; Hald et 8
al., 2004) and co-colonisation of distinct Campylobacter species (Koene et al., 2004; Shen et 9
al., 2001) have been described. However, data on the incidence and extent of genotypic 10
variation of Campylobacter species in companion animals are currently lacking.
11
Such information is important for epidemiological purposes and could help in assessing the 12
role of Campylobacter as a pathogen in these animals. Campylobacter is regarded as a 13
possible cause of diarrhoea in dogs and cats, although prevalence studies, experimental 14
infections and response to antibiotic therapy have been inconclusive (Fox et al., 1983; Hald 15
and Madsen, 1997; Sandberg et al., 2002, Weber et al., 1983a; Weber et al., 1983b). In 16
addition to its clinical relevance, the presence of Campylobacter in companion animals is of 17
interest in view of public health. Contact with pet animals has been shown to be a risk factor 18
for Campylobacter infections in humans and actual transmission has been demonstrated 19
(Wolfs et al., 2001).
20
The presence of multiple strains of a Campylobacter species in a host should be taken in 21
consideration when performing molecular epidemiological and aetiological studies. The aim 22
of this survey is to study the extent of co-colonization of different genotypes of 23
Campylobacter in stool samples from dogs and cats by using AFLP.
24
25
Accepted Manuscript
Material and methods 1
A total of 40 Campylobacter-culture positive faecal samples from dogs (n=34) and cats (n=6) 2
were examined that were either submitted for routine bacteriological analysis (diarrheal 3
animals), samples taken from dogs in a boarding kennel on their day of arrival, and from a 4
veterinary practice (healthy animals). Age distribution of animals was as follows: twenty-two 5
dogs (65%) were adult, 12 dogs (35%) were younger than one year. Cat samples were 6
obtained from one adult, three juveniles, and two for which no information about their age 7
was provided. Diarrhoea was reported in 16 dogs (47%) and 5 cats (83%). The remaining 8
animals had no clinical signs reported by their owner.
9
Samples, with average size of about 25 g (minimum, 5 g), were transported to the laboratory 10
without transport medium or cooling and were collected over a period of two years (August 11
1999-July 2001). The time interval between sampling and processing of the samples ranged 12
from two hours to several days (Table 1a, 1b). Species identification of Campylobacter 13
isolates in some of these samples was reported previously (Koene et al., 2004), and these 14
samples are indicated in Table 1a.
15
Three procedures for the isolation of Campylobacter were used, (i) modified charcoal 16
cefoperazone deoxycholate agar (mCCDA), a blood-free selective agar base supplemented 17
with 32 μg/ml cefoperazone and 10 μg/ml amphotericin (Oxoid CM 739 with Oxoid 18
supplement SR 155), (ii) Karmali, a blood-free charcoal based agar containing 32 μg/ml 19
cefazolin, 20 μg/ml vancomycin, and 100 μg/ml cycloheximide (Oxoid supplement SR 167), 20
and (iii) a filtration method, for which 10 to 12 drops of a fecal suspension in brain heart 21
infusion were placed on a 0.65-μm-pore-size cellulose acetate filter (Sartorius, Goettingen, 22
Germany) on blood agar base no. 2 (Oxoid) supplemented with 5% sheep blood. After 1 h of 23
incubation at 37°C under aerobic conditions, the filter was removed. All plates were 24
incubated in jars under microaerobic conditions (6% O
2, 7% CO
2, 7% H
2and 80% N
2) (Mart,
25
Accepted Manuscript
Anoxomat, Lichtenvoorde, the Netherlands) at 37°C for up to six days. From plates showing 1
growth suspect of Campylobacter (colony morphology and Gram stain), multiple separate 2
colonies (2-33), depending on the number of colonies found in the primary isolation, were 3
subcultured on blood agar base no. 2 (Oxoid) supplemented with 5% sheep blood. These 4
isolates were stored as glycerol stocks at –80°C until further analysis.
5
Chromosomal DNA was isolated using a commercial kit (Promega, Madison, WI, USA.) and 6
DNA concentrations were estimated by agarose gel electrophoresis.
7
Species were identified through PCR-Restriction Fragment Length Polymorphism analysis, 8
using two methods. PCR-RFLP of the 16S rRNA (Marshall et al., 1999) identified C.
9
helveticus, C. hyointestinalis, C. lari and C. upsaliensis. C. jejuni was identified by 10
amplifying a highly polymorphic part of the 23S rRNA gene and subsequently digesting the 11
PCR product with two restriction enzymes, AluI and Tsp509I (Fermer and Engvall, 1999).
12
Campylobacter isolates were genotyped using Amplified Fragment Length Polymorphism 13
analysis (AFLP) as described by Duim (Duim et al., 1999). After electrophoresis, the AFLP 14
patterns were recorded and evaluated with Gene Scan software (PE Applied Bio systems).
15
Densitometric values were transferred to the Bionumerics 3.5 software (Applied Maths, Sint- 16
Martens-Latem, Belgium), and gels were normalized using an internal size standard that was 17
added to each lane. A similarity matrix was created by determining the Pearson product 18
moment correlation coefficient (r). The unweighted pair group method using average linkage 19
(UPGMA) was used to cluster the patterns. A cut-off level of 90% homology was used to 20
discriminate different genotypes.
21 22
Results 23
Statistical analysis of the AFLP results of 451 isolates from 40 Campylobacter positive stools 24
(34 dogs and 6 cats) showed the different Campylobacter species forming distinct clades
25
Accepted Manuscript
correlating with the patterns from reference strains and this was confirmed by the results of 1
the species identification by PCR-RFLP (data not shown).
2
Of the 34 examined dogs, three were shown to be colonized with more than one 3
Campylobacter genotype per species while this was found in four of the six cats. In samples 4
with multiple genotypes, next to a predominant genotype, other genotypes of the same 5
Campylobacter species were present at a lower frequency as identified by AFLP. In canine 6
samples, C. upsaliensis was represented by three AFLP genotypes in sample P1 and by two 7
genotypes in sample N3. C. jejuni showed two different genotypes in one sample (D12) 8
(Table 1a). In four of the six stool samples from cats, strains of distinct Campylobacter 9
species showed variation in genetic fingerprints. This involved C. jejuni (K1, K2) and C.
10
helveticus (K3, K5) (table 1b).
11
A total of 48 unique AFLP genotypes (showing less than 90% homology) were recovered as 12
shown in Figure 1, represented by C. upsaliensis (U1-U25), C. jejuni (J1-J16), C. lari (L1- 13
L2), C. helveticus (He1-He4) and C. hyointestinalis (Hy1). Occasionally, isolates with 14
identical or closely related AFLP patterns (90-100% homology) were isolated from 15
epidemiological unrelated animals as shown in Table 1a and 1b. This included three C. jejuni 16
genotypes (J1, isolated from sample N15 and sample D17; J3, isolated from samples D12 and 17
K2; and J11, from samples N3, N4, N5 and D14) and one C. lari genotype (L1, isolated from 18
samples N5 and N1).
19
The use of multiple isolation methods improved the detection of genetic diversity. In samples 20
with multiple genotypes, variants were distributed over different media, rather than being 21
limited to a single method (data not shown).
22
No correlation was demonstrated between the occurrence of strain variation and clinical 23
status, age of the host or the time interval between sampling and processing of the samples.
24
25
Accepted Manuscript
Discussion 1
We isolated 48 unique Campylobacter genotypes from 40 stool samples (Figure 1). Analysis 2
of the genotypic results indicated that the Campylobacter population in the studied dogs and 3
cats is genetically highly diverse, which is in agreement with studies performed elsewhere 4
(Hald et al., 2004; Moser et al., 2001; Shen et al., 2001; Stanley et al., 1994).
5
Four homologous Campylobacter genotypes were isolated from multiple samples; C. lari 6
genotype 1 (L1) was isolated from two epidemiologically unrelated, clinically healthy dogs.
7
In three cases, homologous genotypes of C. jejuni (J1, J3 and J11) were isolated from 8
epidemiologically unrelated samples. J1 and J11 were isolated from both diarrheic and 9
healthy dogs, while J3 was cultured from two clinical samples from a dog and a cat.
10
Interestingly, in three cases of animals sharing the same household, each individual animal 11
shed Campylobacter with distinct genotypic patterns, or even different Campylobacter 12
species (samples N1+N3, N7+N8 and K2+K3) as is shown in Figure 1. This may indicate that 13
either no animal to animal transmission of strains occurs or that strains alter during prolonged 14
colonization in different individuals.
15
The genetically diverse population shown by most Campylobacter species like C. jejuni, C.
16
coli, C. lari and C. upsaliensis is affected by the occurrence of natural transformation and 17
intragenomic rearrangements. In the present study, in 3 of the 34 (9%) canine samples, more 18
than one genotype of the same Campylobacter species was present, while strain variation was 19
detected in four of the six feline samples. Co-colonization of strains within one host may lead 20
to an increase in the genetic diversity if DNA exchange takes place and new chimeric strains 21
can develop. Both in humans, and in animals like chickens (Boer et al., 2002; Hanninen et al., 22
1999), pigs (Weijtens et al., 1999) and dogs (Hald et al., 2004) time related differences in 23
molecular fingerprints have been described.
24
Accepted Manuscript
It has been shown that host species differ with regard to strain variation within a single host.
1
Mixed infections of Campylobacter in human patients are considered unusual (Richardson et 2
al., 2001; Steinbrueckner et al., 2001) in contrast to the situation in poultry and pigs (Camarda 3
et al., 2000; Jensen et al., 2006; Schouls et al., 2003; Weijtens et al., 1999). However, the 4
mechanisms behind these differences in host-microbe interaction remain largely unidentified.
5
We found no obvious correlation between clinical status and the shedding of multiple strains 6
of Campylobacter. Little data is available on the nature and duration of immunity after 7
Campylobacter colonization in pets, but long term infection and reinfection with different 8
strains without clinical symptoms (Hald et al., 2004; Stanley et al., 1994) in dogs and in cats 9
(Bender et al., 2005) indicates a lack of protective immunity.
10
Although the number of positive cats in this study is limited, the results suggest that 11
Campylobacter strains in feline fecal samples possess a higher degree of genetic diversity 12
than those found in dogs, as strain variation was detected in four of the six samples whereas in 13
canine samples Campylobacter species were generally represented by a single genotype 14
(Table 1a, b). This difference cannot be explained by technical factors such as differences in 15
time interval between sampling and processing of samples or age of the animals. It may 16
indicate that cats are subject to a higher infection pressure than dogs or that there may be 17
differences in host immune responses or colonisation dynamics. Due to the limited number of 18
samples, this warrants further investigation.
19
The detection of genetic diversity was enhanced by the use of multiple isolation methods, but 20
none of the employed methods was superior in detecting different species or genotypes. For 21
routine diagnosis the use of multiple media is not practical and unnecessarily expensive.
22
However, for specific epidemiological studies, the use of multiple media is strongly 23
recommended.
24
25
Accepted Manuscript
Conclusions 1
We conclude that genetic diversity among isolates of C. jejuni and C. upsaliensis within one 2
host occurs, and that the degree may differ in dogs and cats. In all cases in which strain 3
diversity was detected, one genotype dominated, which means that co-colonization can be 4
missed even if multiple colonies are analyzed. This may frustrate epidemiological, 5
aetiological and pathogenetic studies of Campylobacter in dogs and cats, particularly in 6
relation to zoonotic issues.
7 8
Acknowledgements 9
The help of the technicians from the Veterinary Microbiological Diagnostic Center of the 10
Veterinary Faculty is gratefully acknowledged. We also thank Martine Carrière from the 11
Ermelo Animal Clinic for kindly providing us with fecal samples.
12 13
References 14
Bender, J.B., Shulman, S.A., Averbeck, G.A., Pantlin, G.C., Stromberg, B.E., 2005.
15
Epidemiologic features of Campylobacter infection among cats in the upper midwestern 16
United States. J Am Vet Med Assoc 226, 544-547.
17
Boer, P., Wagenaar, J.A., Achterberg, R.P., Putten, J.P., Schouls, L.M., Duim, B., 2002.
18
Generation of Campylobacter jejuni genetic diversity in vivo. Mol Microbiol 44, 351-359.
19
Camarda, A., Newell, D.G., Nasti, R., Di Modugnoa, G., 2000. Genotyping Campylobacter 20
jejuni strains isolated from the gut and oviduct of laying hens. Avian Dis 44, 907-912.
21
Duim, B., Wassenaar, T.M., Rigter, A., Wagenaar, J., 1999. High-resolution genotyping of 22
Campylobacter strains isolated from poultry and humans with amplified fragment length 23
polymorphism fingerprinting. Appl Environ Microbiol 65, 2369-2375.
24
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Fermer, C., Engvall, E.O., 1999. Specific PCR identification and differentiation of the 1
thermophilic campylobacters, Campylobacter jejuni, C. coli, C. lari, and C. upsaliensis. J Clin 2
Microbiol 37, 3370-3373.
3
Fox, J.G., Moore, R., Ackerman, J.I., 1983. Campylobacter jejuni-associated diarrhea in dogs.
4
J Am Vet Med Assoc 183, 1430-1433.
5
Hald, B., Madsen, M., 1997. Healthy puppies and kittens as carriers of Campylobacter spp., 6
with special reference to Campylobacter upsaliensis. J Clin Microbiol 35, 3351-3352.
7
Hald, B., Pedersen, K., Waino, M., Jorgensen, J.C., Madsen, M., 2004. Longitudinal study of 8
the excretion patterns of thermophilic Campylobacter spp. in young pet dogs in Denmark. J 9
Clin Microbiol 42, 2003-2012.
10
Hanninen, M.L., Hakkinen, M., Rautelin, H., 1999. Stability of related human and chicken 11
Campylobacter jejuni genotypes after passage through chick intestine studied by pulsed-field 12
gel electrophoresis. Appl Environ Microbiol 65, 2272-2275.
13
Jensen, A.N., Dalsgaard, A., Baggesen, D.L., Nielsen, E.M., 2006. The occurrence and 14
characterization of Campylobacter jejuni and C. coli in organic pigs and their outdoor 15
environment. Vet Microbiol 116, 96-105.
16
Koene, M.G., Houwers, D.J., Dijkstra, J.R., Duim, B., Wagenaar, J.A., 2004. Simultaneous 17
presence of multiple Campylobacter species in dogs. J Clin Microbiol 42, 819-821.
18
Marshall, S.M., Melito, P.L., Woodward, D.L., Johnson, W.M., Rodgers, F.G., Mulvey, M.R., 19
1999. Rapid identification of Campylobacter, Arcobacter, and Helicobacter isolates by PCR- 20
restriction fragment length polymorphism analysis of the 16S rRNA gene. J Clin Microbiol 21
37, 4158-4160.
22
Moser, I., Rieksneuwohner, B., Lentzsch, P., Schwerk, P., Wieler, L.H., 2001. Genomic 23
heterogeneity and O-antigenic diversity of Campylobacter upsaliensis and Campylobacter 24
helveticus strains isolated from dogs and cats in Germany. J Clin Microbiol 39, 2548-2557.
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Richardson, J.F., Frost, J.A., Kramer, J.M., Thwaites, R.T., Bolton, F.J., Wareing, D.R., 1
Gordon, J.A., 2001. Coinfection with Campylobacter species: an epidemiological problem? J 2
Appl Microbiol 91, 206-211.
3
Sandberg, M., Bergsjo, B., Hofshagen, M., Skjerve, E., Kruse, H., 2002. Risk factors for 4
Campylobacter infection in Norwegian cats and dogs. Prev Vet Med 55, 241-253.
5
Schouls, L.M., Reulen, S., Duim, B., Wagenaar, J.A., Willems, R.J., Dingle, K.E., Colles, 6
F.M., Van Embden, J.D., 2003. Comparative genotyping of Campylobacter jejuni by 7
amplified fragment length polymorphism, multilocus sequence typing, and short repeat 8
sequencing: strain diversity, host range, and recombination. J Clin Microbiol 41, 15-26.
9
Shen, Z., Feng, Y., Dewhirst, F.E., Fox, J.G., 2001. Coinfection of enteric Helicobacter spp.
10
and Campylobacter spp. in cats. J Clin Microbiol 39, 2166-2172.
11
Stanley, J., Jones, C., Burnens, A., Owen, R.J., 1994. Distinct genotypes of human and canine 12
isolates of Campylobacter upsaliensis determined by 16S rRNA gene typing and plasmid 13
profiling. J Clin Microbiol 32, 1788-1794.
14
Steinbrueckner, B., Ruberg, F., Kist, M., 2001. Bacterial genetic fingerprint: a reliable factor 15
in the study of the epidemiology of human Campylobacter enteritis? J Clin Microbiol 39, 16
4155-4159.
17
Weber, A., Lembke, C., Schafer, R., Seifert, U., Berg, H., 1983a. Demonstration of 18
Campylobacter jejuni in fecal samples from dogs. Berl Munch Tierarztl Wochenschr 96, 232- 19
234.
20
Weber, A., Schafer, R., Lembke, C., Seifert, U., 1983b. Occurrence of Campylobacter jejuni 21
in cats. Berl Munch Tierarztl Wochenschr 96, 48-50.
22
Weijtens, M.J., Reinders, R.D., Urlings, H.A., Van der Plas, J., 1999. Campylobacter 23
infections in fattening pigs; excretion pattern and genetic diversity. J Appl Microbiol 86, 63- 24
70.
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Wolfs, T.F., Duim, B., Geelen, S.P., Rigter, A., Thomson-Carter, F., Fleer, A., Wagenaar, 1
J.A., 2001. Neonatal sepsis by Campylobacter jejuni: genetically proven transmission from a 2
household puppy. Clin Infect Dis 32, E97-99.
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Table 1a Genotyping results by AFLP from Campylobacter positive samples from dogs
1
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Table 1b Genotyping results by AFLP from Campylobacter positive samples from cats
1
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Figure legends 1
2
Figure 1. Representative AFLP patterns of Campylobacter species from individual samples 3
isolated from fecal samples of dogs and cats. The black line indicates the cut-off level (less 4
than 90 % genetic similarity) for pattern differentiation. Epidemiologically related animals - 5
by sharing the same household - are indicated by symbols (■,▼,‡).
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Table 1a Genotyping results by AFLP from Campylobacter positive samples from dogs
C. upsaliensis C. jejuni C. lari C. hyointestinalis
P1 11 U21 (3), U22 (1), U25 (6) Hy1 (1) 8 no 9 months
N3* 11 U1 (6), U2 (1) J11 (3) L2 (1) 4 no 3 years
N5* 18 U15 (14) J11 (3) L1 (1) 4 no 10 years
N4* 10 U18 (1) J11 (9) 4 no 2 years
N9* 23 U4 (18) J2 (5) 4 no 4 years
N8* 19 U16 (19) 4 no 4 years
N10* 18 U7 (18) 4 no 7 months
N11* 7 U8 (7) 2 no 9 months
N12* 9 U17 (9) 2 no 3 years
P2 12 U20 (12) 8 no 9 months
D2 27 U19 (27) 12 yes 3 years
D4 13 U14 (13) 36 yes 4 months
D9 17 U9 (17) 36 yes 1 year
D10 2 U10 (2) 36 yes 3 years
D11 12 U11 (12) 12 yes 3 months
D13 33 U24 (33) 36 yes 6 months
D15* 5 U13 (5) 36 yes 5 years
D16* 3 U23 (3) 36 yes 3 months
G57* 9 U12 (9) 60 no adult
G61* 4 U5 (4) 108 yes adult
G70* 6 U6 (6) 60 no adult
D12 9 J3 (8), J10 (1) 36 yes 5 months
N2* 2 J16 (2) 4 no 7 years
N7* 8 J11 (8) 4 no 4 years
N15* 2 J1 (2) 6 no 7 months
D3 2 J7 (2) 60 yes 10 years
D6 12 J5 (12) 84 yes 2 months
D7 21 J6 (21) 12 yes 1 year
D8 24 J8 (24) 12 yes 1 year
D14 4 J11 (4) 84 yes 6 months
D17* 8 J1 (8) 6 yes adult
G50-1 6 J4 (6) 60 no adult
G48* 7 J12 (7) 108 no adult
N1* 2 L1 (2) 4 no 2 years
* Speciation of the isolates by PCR is described in Koene et al, 2004
time interval sampling- processing
(hours)
diarrhoea age sample ID
No of isolates
tested
AFLP-type isolated (number of isolates)
Table 1a
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Table 1b Genotyping results by AFLP from Campylobacter positive samples from cats
C. upsaliensis C. jejuni C. helveticus
G44-1
6
U3 (6) 48 no adultK1
21
J13 (1), J14 (20) 36 yes 6 weeksK2
13
J3 (11), J9 (2) 12 yes not reportedK7
21
J15 (21) 36 yes 3 monthsK3
2
He1 (1), He4 (1) 12 yes not reportedK5
12
He2 (1), He3 (11) 12 yes 3 monthsdiarrhoea age
sample ID No of isolates tested
AFLP-type isolated (number of isolates) time interval sampling- processing
(hours)
Table 1b
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Pearson correlation (Opt:1.00%) [0.0%-100.0%]
AFLP ABI373 GC
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90
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40
30
20
10
AFLP ABI373 GC
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C.
jejuni jejuni jejuni jejuni jejuni jejuni jejuni jejuni jejuni jejuni jejuni jejuni jejuni jejuni jejuni jejuni helveticus helveticus helveticus helveticus hyointestinalis lari lari upsaliensis upsaliensis upsaliensis upsaliensis upsaliensis upsaliensis upsaliensis upsaliensis upsaliensis upsaliensis upsaliensis upsaliensis upsaliensis upsaliensis upsaliensis upsaliensis upsaliensis upsaliensis upsaliensis upsaliensis
D17.4 N9.10 K2.8 G50Dj6 D6.5 D7.10 D3.2 D8.17 K2.2 D12.24 N7.9 G48.3 K1.5 K1.14 K7.5 N2.1 K3.2 K5.1 K5.13 K3.1 P1.5 N1.15 N3.13 N3.2 N3.15 G44.5 N9.9 G61.3 G70.3 N10.3 N11.3 D9.10 D10.1 D11.5 G57.4 D15.2 D4.7 N5.2 N8.1 N12.5 N4.1 D2.10 P2.7
J1 J2 J3 J4 J5 J6 J7 J8 J9 J10 J11 J12 J13 J14 J15 J16 He1 He2 He3 He4 Hy1 L1 L2 U1 U2 U3 U4 U5 U6 U7 U8 U9 U10 U11 U12 U13 U14 U15 U16 U17 U18 U19 U20
species sample genotype
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