PCR ribotyping of Clostridium difficile isolates originating from human and animal sources

Arroyo, Luis; Kruth, Stephen A.; Willey, Barbara M.; Staempfli, H. R.; D, Low; Weese, J. Scott

doi:10.1099/jmm.0.45805-0

Cited by 119 publications

(63 citation statements)

References 23 publications

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“…In vitro toxin production results from our limited sample suggest that toxinotype V strains have the potential to cause increased severity of human disease, although further studies are needed to corroborate this association. Although they share similar clinical features, evidence suggests that the predominant strains causing CDAD in humans and different animal species are distinct (8,38). Nonetheless, our fi nding of similarity between relatively widespread animal strains of C. diffi cile and strains responsible for occasional human disease raises the possibility of interspecies transmission.…”

Section: Discussionmentioning

confidence: 40%

Toxinotype VClostridium difficilein Humans and Food Animals

Jhung

Thompson

Killgore

et al. 2008

Emerg. Infect. Dis.

197

155

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Clostridium diffi cile is a recognized pathogen in neonatal pigs and may contribute to enteritis in calves. Toxinotype V strains have been rare causes of human C. diffi cileassociated disease (CDAD). We examined toxinotype V in human disease, the genetic relationship of animal and human toxinotype V strains, and in vitro toxin production of these strains. From 2001 through 2006, 8 (1.3%) of 620 patient isolates were identifi ed as toxinotype V; before 2001, 7 (<0.02%) of ≈6,000 isolates were identifi ed as toxinotype V. Six (46.2%) of 13 case-patients for whom information was available had community-associated CDAD. Molecular characterization showed a high degree of similarity between human and animal toxinotype V isolates; all contained a 39-bp tcdC deletion and most produced binary toxin. Further study is needed to understand the epidemiology of CDAD caused by toxinotype V C. diffi cile, including the potential of foodborne transmission to humans. R ecent evidence suggests that the epidemiology of Clostridium diffi cile-associated disease (CDAD) is increasing in incidence and severity (1-3). These changes are due, at least in part, to the emergence of a more virulent C. diffi cile strain, designated NAP1 (based on its pulsed-fi eld gel electrophoresis [PFGE] pattern), BI (by restriction endonuclease analysis [REA]), toxinotype III (by PCR characterization of the pathogenicity locus), and 027 (by PCR ribotyping) (4). However, the emergence of BI/NAP1/027 may not be solely responsible for changes in CDAD epidemiology, and the origin of this and other virulent strains is still largely unknown. C. diffi cile has also recently emerged as a pathogen or commensal in food animals such as neonatal pigs and beef and dairy calves (5-7); most of these animal isolates are toxigenic. Although several ribotypes have been identifi ed in calves, the predominant ribotype in both calves and pigs is a toxinotype V strain (8,9). Moreover, recent reports suggest that C. diffi cile strains recognized as causes of human disease may contaminate retail meats (10).To better understand whether food sources could be a source of infection for humans, we investigated recent and past human CDAD caused by toxinotype V C. diffi cile and compared isolates from these cases with those recovered from neonatal pigs and calves. We documented apparent changes in the frequency with which these toxinotype V strains cause human CDAD and compared the molecular characterization and toxin production of these strains with those of recent epidemic (i.e., BI/NAP1/027) and nonepidemic isolates. Materials and Methods Human Case Finding and Defi nitionsCase fi nding was performed by reviewing recent and past human isolates of interest from 2 sources. Cases were defi ned as patients with clinical isolates identifi ed as toxinotype V by analysis of restriction fragment length polymorphisms (RFLPs) of toxin-encoding genes. First, we reviewed 620 C. diffi cile human isolates sent to the Centers for Disease Control and Prevention (CDC) from healthcare facilities and ...

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Section: Discussionmentioning

confidence: 40%

Toxinotype VClostridium difficilein Humans and Food Animals

Jhung

Thompson

Killgore

et al. 2008

Emerg. Infect. Dis.

197

155

View full text Add to dashboard Cite

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“…4) (41). Patients with CDI acquire the organism from the environment (31) and interspecies transmission of C. difficile may be possible (4). Because the spores of C. difficile are heat resistant, a role for food (especially meats) in the transmission of C. difficile may be possible.…”

Section: Resultsmentioning

confidence: 99%

Microarray Identification of Clostridium difficile Core Components and Divergent Regions Associated with Host Origin

et al. 2009

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Clostridium difficile is a gram-positive, spore-forming enteric anaerobe which can infect humans and a wide variety of animal species. Recently, the incidence and severity of human C. difficile infection has markedly increased. In this study, we evaluated the genomic content of 73 C. difficile strains isolated from humans, horses, cattle, and pigs by comparative genomic hybridization with microarrays containing coding sequences from C. difficile strains 630 and QCD-32g58. The sequenced genome of C. difficile strain 630 was used as a reference to define a candidate core genome of C. difficile and to explore correlations between host origins and genetic diversity. Approximately 16% of the genes in strain 630 were highly conserved among all strains, representing the core complement of functional genes defining C. difficile. Absent or divergent genes in the tested strains were distributed across the entire C. difficile 630 genome and across all the predicted functional categories. Interestingly, certain genes were conserved among strains from a specific host species, but divergent in isolates with other host origins. This information provides insight into the genomic changes which might contribute to host adaptation. Due to a high degree of divergence among C. difficile strains, a core gene list from this study offers the first step toward the construction of diagnostic arrays for C. difficile.

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“…This Gram-positive, spore-forming, anaerobic bacterium also causes infections in a number of animal species, especially horses, piglets, calves, and dogs (2). In humans, the clinical symptoms range from mild watery diarrhea and abdominal pain to fulminant pseudomembranous colitis and are caused by two main exotoxins, TcdA and TcdB, encoded by the pathogenicity locus of toxigenic isolates.…”

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confidence: 99%

Characterization of Temperate Phages Infecting Clostridium difficile Isolates of Human and Animal Origins

Sekulović

Garneau

Néron

et al. 2014

Appl Environ Microbiol

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Clostridium difficile is a Gram-positive pathogen infecting humans and animals. Recent studies suggest that animals could represent potential reservoirs of C. difficile that could then transfer to humans. Temperate phages contribute to the evolution of most bacteria, for example, by promoting the transduction of virulence, fitness, and antibiotic resistance genes. In C. difficile, little is known about their role, mainly because suitable propagating hosts and conditions are lacking. Here we report the isolation, propagation, and preliminary characterization of nine temperate phages from animal and human C. difficile isolates. Prophages were induced by UV light from 58 C. difficile isolates of animal and human origins. Using soft agar overlays with 27 different C. difficile test strains, we isolated and further propagated nine temperate phages: two from horse isolates (CD481-1 and CD481-2), three from dog isolates (CD505, CD506, and CD508), and four from human isolates (CD24-2, CD111, CD146, and CD526). Two phages are members of the Siphoviridae family (CD111 and CD146), while the others are Myoviridae phages. Pulsed-field gel electrophoresis and restriction enzyme analyses showed that all of the phages had unique double-stranded DNA genomes of 30 to 60 kb. Phages induced from human C. difficile isolates, especially the members of the Siphoviridae family, had a broader host range than phages from animal C. difficile isolates. Nevertheless, most of the phages could infect both human and animal strains. Phage transduction of antibiotic resistance was recently reported in C. difficile. Our findings therefore call for further investigation of the potential risk of transduction between animal and human C. difficile isolates.

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PCR ribotyping of Clostridium difficile isolates originating from human and animal sources

Cited by 119 publications

References 23 publications

Toxinotype VClostridium difficilein Humans and Food Animals

Toxinotype VClostridium difficilein Humans and Food Animals

Microarray Identification of Clostridium difficile Core Components and Divergent Regions Associated with Host Origin

Characterization of Temperate Phages Infecting Clostridium difficile Isolates of Human and Animal Origins

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