Clostridium difficile Genotypes in Piglet Populations in Germany

Abstract

dClostridium difficile was isolated from 147 of 201 (73%) rectal swabs of piglets from 15 farms of Lower Saxony and North Rhine-Westphalia. In 14 farms, 14 to 100% (mean, 78%) of the animals tested were culture positive. The rate of isolation was 68% postpartum, increased to 94% in animals 2 to 14 days of age, and declined to 0% for animals 49 days of age and older. There was no link between isolation and antibiotic treatment or diarrhea of piglets. Strains were assigned to 10 PCR ribotypes, and up to 4 PCR ribotypes were found to be present at the same time on a farm. The closely related PCR ribotypes 078 (55%) and 126 (20%) were most frequently recovered and were present in 13 of the 14 positive farms. The comparison of multilocus VNTR (variable number of tandem repeats) analysis (MLVA) data from this study and previously published data on human, porcine, and bovine PCR ribotype 078 isolates from 5 European countries revealed genetic differences between strains of different geographic origin and confirmed the relatedness of human and porcine C. difficile isolates. This study demonstrated that the human-pathogenic PCR ribotypes 078 and 126 are predominant in piglets in Germany. The results suggest that presence of C. difficile is correlated with animal age but not with antibiotic treatment or clinical disease. MLVA indicated that strains of the same geographical origin are often genetically related and corroborated the hypothesis of a close epidemiological connection between human and porcine C. difficile isolates.

Nationwide Surveillance Study of Clostridium difficile in Australian Neonatal Pigs Shows High Prevalence and Heterogeneity of PCR Ribotypes

“…C. difficile prevalence in piglets aged less than 7 days was 67.2%. This is higher than reported in the United States (29.6%) (6), Slovenia (50.9%) (7), and the Czech Republic (56.7%) (8) and similar to recent reports from Sweden (67.2%) (11) and Germany (73%) (24). Recovery of C. difficile by enrichment culture was significantly better than direct culture on ChromID (52.4% versus 67.2%; P ϭ 0.001), in accordance with studies of human CDI (25).…”

DNA Microarray-Based PCR Ribotyping of Clostridium difficile

“…Knetsch et al (27) found that the PCR ribotypes 078, 126, 033, and 045 are highly related in MLST, as they displayed the same sequence type, ST-11, and contained a deletion (⌬39 bp) in the tcdC gene. Recently published data on the sequence similarities of conserved genes (e.g., gyrA, tcdB, and RNA polymerases) in C. difficile PCR ribotypes showed that strains of ribotypes 033, 078, and 126 were very similar, forming a distinct ribotype group or cluster (19,20,28). Accordingly, the results of the array typing confirm the relatedness of ribotypes assigned to the 033-045-078 group.…”

“…A collection of 168 C. difficile reference, field, and type strains representing 48 seq-PCR ribotypes was used to validate the microarraybased assay (Table 1). Eighty-six field strains isolated from animal (calf, piglet, cat, and dog) and human (2 strains) sources were obtained from previous investigations (18)(19)(20) …”

“…Bacterial culture, genomic DNA extraction, toxin gene detection, and seq-PCR ribotyping were performed as previously described (20).…”

Investigation of Clostridium difficile interspecies relatedness using multilocus sequence typing, multilocus variable-number tandem-repeat analysis and antimicrobial susceptibility testing

“…These were considered clonal by MLVA, but in this case the isolates were obtained on two different sampling days and the animals proceeded from different herds. As previously suggested, this finding may indicate either an epidemiological connection between farms (Scheeberg et al, 2013) or slaughterhouse contamination. However, clonal strains were not detected among C. difficile isolates from pig and cattle origins, which indicates that crosscontamination between the two slaughter lines (bovine and porcine) during processing is unlikely.…”