A novel PCR-based restriction fragment length polymorphism analysis of the Brachyspira nox gene was developed. The restriction patterns for Brachyspira hyodysenteriae, B. pilosicoli, B. intermedia, B. murdochii, and B. innocens were highly distinct with two restriction endonucleases only. The assay proved to be user-friendly and robust.The porcine intestinal tract is frequently colonized by different Brachyspira species. Brachyspira hyodysenteriae is the causative agent of swine dysentery, and B. pilosicoli causes intestinal spirochetosis (9). The other species, namely, B. intermedia, B. murdochii, and B. innocens, are considered nonpathogenic (9). Due to differences in pathogenicity (9) and the zoonotic potential of B. pilosicoli (15), rapid and user-friendly differentiation between species is important with respect to the economic use of medication, preharvest food safety, and animal trade.Currently, porcine Brachyspira isolates are differentiated based on phenotypic criteria, including intensity of hemolysis, indole production, hippurate hydrolysis, and activities of ␣-galactosidase, ␣-glucosidase, and -glucosidase (6, 14). In addition, a positive ring phenomenon is indicative of B. hyodysenteriae (11). Due to variable results in these tests and difficulty in correctly assessing the intensity of hemolysis, it seems desirable to replace phenotypic identification schemes with genotypic methods. The currently used sequencing analyses of 16S or 23S ribosomal DNA (rDNA), the nox gene, or unidentified DNA segments or PCR methods based on these data (1,4,5,10,12,13) either are not practical for routine veterinary diagnostic purposes or are confined to the identification of selected species only. Therefore, the purpose of the present study was to develop a robust and user-friendly method for the identification of all porcine Brachyspira species and to evaluate its performance in routine diagnostic work.
Transient bacteraemia of oral origin commonly occurs during dental extraction in horses. As none of the horses developed complications associated with bacteraemia during the observation period after surgery, the significance of this bacteraemia remains uncertain. The Summary is available in Chinese - see Supporting Information.
Swine dysentery (SD) is a mucohaemorrhagic colitis of grower/finisher pigs classically resulting from infection by the anaerobic intestinal spirochaete Brachyspira hyodysenteriae. This study aimed to determine whether B. hyodysenteriae isolates from pigs in three healthy German multiplier herds supplying gilts to other farms differed from isolates from nine German production herds with SD. Isolates were subjected to whole genomic sequencing, and in silico multilocus sequence typing showed that those from the three multiplier herds were of previously undescribed sequence types (ST132, ST133 and ST134), with all isolates from the same herd having the same ST. All isolates were examined for the presence of 332 genes encoding predicted virulence or virulence lifestyle associated factors, and these were well conserved. Isolates from one multiplier herd were atypical in being weakly haemolytic: they had 10 amino acid substitutions in the haemolysin III protein and five in the haemolysin activation protein compared to reference strain WA1, and had a disruption in the promoter site of the hlyA gene. These changes likely contribute to the weakly haemolytic phenotype and putative lack of virulence. These same isolates also had nine base pair insertions in the iron metabolism genes bitB and bitC and lacked five of six plasmid genes that previously have been associated with colonisation. Other overall differences between isolates from the different herds were in genes from three of five outer membrane proteins, which were not found in all the isolates, and in members of a block of six plasmid genes. Isolates from three herds with SD had all six plasmid genes, while isolates lacking some of these genes were found in the three healthy herds—but also in isolates from six herds with SD. Other differences in genes of unknown function or in gene expression may contribute to variation in virulence; alternatively, superior husbandry and better general health may have made pigs in the two multiplier herds colonised by “typical” strongly haemolytic isolates less susceptible to disease expression.
Streptococcus suis is an economically important pathogen of pigs as well as a zoonotic cause of human disease. Serotyping is used for further characterization of isolates; some serotypes seem to be more virulent and more widely spread than others. This study characterizes a collection of German field isolates of Streptococcus suis from pigs dating from 1996 to 2016 with respect to capsular genes (cps) specific for individual serotypes and pathotype by multiplex PCR and relates results to the clinical background of these isolates. The most prominent finding was the reduction in prevalence of serotype-2/serotype-1/2 among invasive isolates during this sampling period, which might be attributed to widely implemented autogenous vaccination programs in swine against serotype 2 in Germany. In diseased pigs (systemically ill; respiratory disease) isolates of serotype-1/serotype-14, serotype-2/serotype-1/2, serotype 3 to 5 and 7 to 9 were most frequent while in carrier isolates a greater variety of cps types was found. Serotype-1/serotype-14 seemed to be preferentially located in joints, serotype 4 and serotype 3 in the central nervous system, respectively. The virulence associated extracellular protein factor was almost exclusively associated with invasive serotype-1/serotype-14 and serotype-2/serotype-1/2 isolates. In contrast, lung isolates of serotype-2/serotype-1/2 mainly harbored the gene for muramidase-released protein. Serotype 4 and serotype 9 isolates from clinically diseased pigs most frequently carried the muramidase-released protein gene and the suilysin gene. When examined by transmission electron microscopy all but one of the isolates which were non-typable by molecular and serological methods showed various amounts of capsular material indicating potentially new serotypes among these isolates. Given the variety of cps types/serotypes detected in pigs, not only veterinarians but also medical doctors should consider other serotypes than just serotype 2 when investigating potential human cases of Streptococcus suis infection.
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