Abstract Book of EAVLD2012 - eavld congress 2012

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DETECTION OF BRACHYSPIRA HYODYSENTERIAE AND BRACHYSPIRA PILOSICOLI IN SWISS PIGS
USING A COMBINATION OF CULTURE AND PCR
Sarah Prohaska 1 , Helen Huber 1 , Titus Sydler 2 , Max M. Wittenbrink 1
1 Institute of Veterinary Bacteriology, Vetsuisse Faculty, University of Zurich, Switzerland
2
Institute of Veterinary Pathology, Vetsuisse Faculty, University of Zurich, Switzerland
Introduction
Swine dysentery and porcine intestinal spirochaetosis caused by
pathogenic Brachyspira (B.) spp. (B. hyodysenteriae and
B. pilosicoli) lead to considerable economic loss and are
therefore of increasing importance to the pig industry.
B. hyodysenteriae colonise the epithelium of the large intestine
and subsequently induce severe mucohaemorrhagic colitis
showing the typical signs of diarrhoea containing blood and plugs
of mucus (1). Infections with B. pilosicoli however are
characterised by moderate growth loss and mild diarrhoea (2).
The goal of this study was to evaluate the presence of
B. hyodysenteriae and B. pilosicoli in Swiss pig herds by a
combination of culture and PCR.
Swine, Brachyspira spp., culture, PCR
Materials & methods
Over a time period of three years (2009-2011) ligated colon
sections (n=140) from dissected pigs as well as faecal swabs
(n= 661) collected on 228 Swiss pig farms were subjected to a
cultural assay and subsequent PCR for the detection of
B. hyodysenteriae and B. pilosicoli. Faecal swabs were taken
from pigs suffering from acute diarrhoea.
For culture, the selective BJ-Agar (Trypticase soy agar
supplemented with 5% cattle blood and Colistin, Vancomycin,
Spectinomycin, Spriramycin and Rifampicin) was used (3). Plates
were incubated under anaerobic conditions at 42°C for 4-6 days.
To evaluate the presence of Brachyspira spp., native
preparations of colony material were investigated by darkfield
microscopy. If spirochaetes were found, DNA was extracted from
suspicious colonies. Subsequently, PCR using two primer pairs
for the detection of B. hyodysenteriae and B. pilosicoli was
performed (4).
Results
In two of the 140 (1.4%) tested colon sections B. hyodysenteriae
could be detected, whereas B. pilosicoli was not found in these
samples (Figure 1).
In terms of faecal swabs, cultivation and subsequent confirmation
by PCR yielded B. hyodysenteriae in 122 (18.5%) samples and
B. pilosicoli in 33 samples (5.0%). However, analysis of seven
(1.0%) swabs revealed a co-infection of B. hyodysenteriae and
B. pilosicoli (Figure 2).
Figure 2: Results of faecal swabs
Discussion & conclusions
The combination of culture and PCR for diagnosis of Swine
dysentery and intestinal spirochaetosis proofed to be a
successful tool.
The results of this study show a remarkable presence of
pathogenic Brachyspira spp. in Switzerland. B. hyodysenteriae
was detected in samples taken from pigs from 12 of 26 cantons
in Switzerland. Due to this situation, the Swiss pig health service
implemented a monitoring system of pathogenic Brachyspira spp.
starting 2012.
References
1. Hampson D.J., Atyeo R.F., Combs B.G., 1997. Swine Dysentery 175-
209. In: Intestinal Spirochaetes in Domestic Animals and Humans
(Hampson D.J. and Stanton T.B.), CAB International, Wallingford, UK.
2. Taylor D.J., Simmons J.R., Laird H.M., 1980. Production of diarrhoea
and dysentery in pigs by feeding pure cultures of a spirochaete differing
from Treponema hyodysenteriae. Vet. Rec. 106, 326-332.
3. Dünser M., Schweighardt H., Pangerl R., Awad-Masalmeh M., Schuh
M., 1997. Schweinedysenterie und Sprirochaetendiarrhoe – vergleichende
Untersuchungen serpulinenbedingter Enteritiden. Wien. Tierärztl. Mschr.
84, 151-161.
4. La T., Philips N.D., Hampson D.J. 2003. Development of a Duplex PCR
Assay for Detection of Brachyspira hyodysenteriae and Brachyspira
pilosicoli in Pig Feces. J.Clin. Microbiol. 41, 3372-3375.
Figure 1: Results of ligated colon samples