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- aggregation classification "C3".
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- aggregation date "2013".
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- aggregation language "eng".
- aggregation subject "Veterinary Sciences".
- aggregation title "Importance of microbial culture to identify (novel) highly beta-hemolytic Brachyspira species".
- aggregation abstract "Brachyspira hyodysenteriae is considered the primary etiological agent of swine dysentery, causing severe mucohemorrhagic diarrhea. Up till now, the presence of strong beta-hemolysis and ring-phenomenon differentiates the highly pathogenic B. hyodysenteriae from weak hemolytic - and less pathogenic - species (B. innocens, B.murdochii and B. intermedia). However, recent data suggest that also other strongly beta-hemolytic Brachyspira species, identified as B. intermedia, can induce a dysentery-like disease in pigs (1). In a routine monitoring protocol, two gilts were presented for necropsy. Soft, watery colonic content was found. Microbial culture of fecal content revealed strongly beta-hemolytic, ring phenomenon-positive spirochetes, indicative for B. hyodysenteriae. In additional samples taken at the farm, strongly beta-hemolytic Brachyspira isolates, with ring phenomenon were again found on microbial culture, whereas PCR analysis (ADIAVET BRACHY) did not confirm the presence of B. hyodysenteriae. In order to elucidate this discrepancy, additional PCR’s were performed targeting the 16S rRNA, 23S rRNA, nox, hlyA and tlyA genes of different swine-related Brachyspira spp (2,4). PCR’s targeting nox and 23s rRNA specific for B. intermedia were positive. However, all isolates were also positive for tlyA and hlyA gene, hemolysins considered being virulence factors of B. hyodysenteriae. The hlyA gene was properly located between fabG en fabF genes encoding an ACP-reductase and –synthetase(4). A sequencing protocol, targeting a highly variable region of the Brachyspira nox gene(3), was performed, identifying the isolates as B. intermedia. Additionally, the obtained sequence showed high (>99%) similarity with sequences of previously published strongly hemolytic B. intermedia isolates(1). This case report highlights the fact that strongly hemolytic isolates, belonging to the “less pathogenic” Brachyspira species can be isolated from field cases in Europe. While the presence of strong beta-hemolysis and ring phenomenon might indicate the pathogenicity (1), intense genotypic testing is required to correctly identify these isolates to the species level. References 1. Burrough et al. (2012) Comparative virulence of clinical Brachyspira spp. isolates in inoculated pigs. Journal of Veterinary Diagnostic Investigation 24(6) 1025-1034. 2. Backhäns et al. (2010) Phenotypic and molecular characterization of Brachyspira spp isolated from wild rodents. Environmental Microbiology Reports 2(6) 720–727. 3. Weissenböck et al. (2005) Amplification and sequencing of Brachyspira spp. specific portions of nox using paraffin-embedded tissue samples from clinical colitis in Austrian pigs shows frequent solitary presence of Brachyspira murdochii. Veterinary Microbiology 111(1-2) 67-75. 4. Barth et al. (2012) Demonstration of genes encoding virulence and virulence life-style factors in Brachyspira spp. isolates from pigs. Veterinary Microbiology 155(2-4) 438-434.".
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