Ribotypes Research Articles

Diversity of Listeria Ribotypes Recovered from Dairy Cattle, Silage, and Dairy Processing Environments

Listeria strains isolated over the past 10 years from farms and dairy processing environments were subjected to strain-specific ribotyping using the automated Riboprinter microbial characterization system, alpha version (E. I. du Pont de Nemours & Co., Inc.). A total of 388 Listeria isolates from 20 different dairy processing facilities were examined along with 44 silage, 14 raw milk bulk tank, and 29 dairy cattle (26 udder quarter milk, 1 brain, 1 liver, and 1 aborted fetus) isolates. These 475 isolates included 93 L. monocytogenes, 362 L. innocua, 11 L. welshimeri, 6 L. seeligeri, 2 L. grayi, and 1 L. ivanovii strains. Thirty-seven different Listeria ribotypes (RTs) comprising 16 L. monocytogenes (including five known clinical RTs responsible for foodborne listeriosis), 12 L. innocua, 5 L. welshimeri, 2 L. seeligeri, 1 L. ivanovii, and 1 L. grayi were identified. Greatest diversity was seen among isolates from dairy processing facilities with 14 of 16 (87.5%) of the L. monocytogenes RTs (including five clinical RTs) and 19 of 21 (90.5%) of the non-L. monocytogenes RTs detected. Sixty-five of the 93 L. monocytogenes isolates belonged to a group of five clinical RTs. These five clinical RTs included one RT unique to dairy processing environments, two RTs common to dairy processing environments and silage, and one RT common to dairy processing environments, silage, and dairy cattle with the last RT appearing in dairy processing environments, silage, raw milk bulk tanks, and dairy cattle. These findings, which support the link between on-farm sources of Listeria contamination (dairy cattle, raw milk, silage) and subsequent contamination of dairy processing environments, stress the importance of farm-based HACCP programs for controling listeriae.

Identification of IS1356, a new insertion sequence, and its association with IS402 in epidemic strains of Burkholderia cepacia infecting cystic fibrosis patients.

Burkholderia cepacia is now recognized as an important opportunistic pathogen in cystic fibrosis (CF) and other compromised patients. Epidemicity among CF patients has been attributed to at least one particularly infectious strain (strain ET12), and both genetic evidence and anecdotal evidence suggest that this strain, currently endemic in Ontario, and those causing an epidemic in the United Kingdom, are indeed the same. Our study was conducted to determine whether there was any association between the presence of various insertion sequence (IS) elements, the cable pilin subunit gene (cblA), electrophoretic type (ET), and ribotype (RT) in a collection of 97 clinical and 2 environmental isolates of B. cepacia. No apparent linkage was found for IS elements IS401, IS402, IS406, IS407, and IS408 with ET or RT. The cblA target, said to be a marker for high infectivity, was detected in 100% (38 of 38) of strains of B. cepacia ET12 and in a single strain of ET13 that differed in a single enzyme allele. A new IS, IS1356, identified during the investigation, was present in 71.7% of all isolates, and 50.7% of these isolates harbored IS1356 as a hybrid IS element inserted into IS402. IS1356 is 1,353 bp in length, and when it is inserted into IS402 it results in a 10-bp duplication at the site of insertion. IS1356 contains one major open reading frame of 1,260 bp coding for a putative transposase which has significant homology to ISRm3 in Rhizobium meliloti (59%) and to an undesignated IS element in Corynebacterium diphtheriae (49%). The IS402-IS1356 element was found exclusively in the epidemic strains from Ontario and the United Kingdom, being detected in 94.7% (36 of 38 isolates) of B. cepacia ET12 isolates. Of the two ET12 isolates found to be devoid of the IS402-IS1356 element, both contained IS1356 unassociated with IS402, one was temporally unrelated to the epidemic, and the other was from a CF patient in a geographic area remote from Ontario and the United Kingdom. It is evident that the IS402-IS1356 hybrid element, the cblA pilin subunit gene, and the allelic suite represented by multilocus enzyme electrophoretic type ET12 may provide useful markers for the epidemic, highly transmissible transatlantic strain isolated in Ontario and the United Kingdom.

Molecular polymorphism of the lyme disease agent Borrelia garinii in northern Europe is influenced by a novel enzootic Borrelia focus in the North Atlantic.

Lyme disease Borrelia species are distributed in temperate areas of North America and Eurasia. To elucidate the distribution of borreliae in subarctic regions, strains isolated from Ixodes ricinus and Ixodes uriae ticks found on islands in the northern Atlantic and Baltic Sea were molecularly characterized. All isolates were verified as Borrelia garinii by 16S rRNA gene analysis and immunoblotting with monoclonal antibodies specific for the outer surface proteins A and C. Three ribotypes (RTs) of B. garinii were delineated. I. ricinus complex-associated RT1 was phenotypically most heterogeneous. Two newly identified ribotypes were shared by different tick species and conformed to two established OspA serotypes. RT2 was restricted to the islands in the northern Baltic Sea, whereas RT3 was recovered also from ticks found in the North Atlantic. In conclusion, molecular polymorphism of the studied borrelia isolates suggests a complex enzootic potential of B. garinii in northern Europe and implies a novel, seabird tick I. uriae-associated enzootic focus of Lyme disease borreliae in the North Atlantic.

Linkage analysis of geographic and clinical clusters in Pseudomonas cepacia infections by multilocus enzyme electrophoresis and ribotyping.

Multilocus enzyme electrophoresis and ribotyping were used to characterize 83 strains of Pseudomonas cepacia, mostly isolated from cystic fibrosis (CF) patients, although a number of isolates from non-CF nosocomial infections and reference environmental strains were represented. Twenty enzyme electrophoretic types (ETs) were determined; of these, one clone (ET12) was associated with six of nine ribotypes (RTs) said to be geographically representative of the United Kingdom and all of the Ontario (Canada) isolates from CF patients. This clone was not associated with nosocomial infections or environmental strains and was never found in CF isolates from British Columbia or Nova Scotia, Canada, or a center in the eastern United States. Individual isolate EcoRI RT signatures did not cluster geographically as did the ET signatures by clonal analysis. Frequently RTs occurred in more than a single ET. Known point source focal nosocomial outbreaks were typified by single ETs and stable RTs. Dendrographic analysis of the strains grouped those strains from CF patients, nosocomial outbreaks, and environmental sources into separate ET families, and diversity analysis indicated that, with the exception of ET17, CF isolates clustered in unique and closely related ETs different from those from nosocomial and environmental sources. This study has also shown the potential of multilocus enzyme electrophoresis to monitor the intercontinental spread of P. cepacia strains in CF patients, and this may have a significant impact on plans for CF patient summer camps and design of infection control practices. Whether the intercontinental ET12 clone, which predominates in the United Kingdom and the province of Ontario, linked by summer camp acquisition, has increased virulence for CF patients remains to be established.

СОВРЕМЕННЫЕ СТРАТЕГИИ ДИАГНОСТИКИ И ЛЕЧЕНИЯ CLOSTRIDIUM DIFFICILE-ИНФЕКЦИИ (ОБЗОР ЛИТЕРАТУРЫ)

The incidence and mortality rate of C. difficile infection (CDI) has increased remarkably over the last 10-20 years. CDI is a leading cause of healthcare-associated infections and antibiotic-associated diarrhoea. Increasing incidence and severity of CDI may be related with frequent antibiotic use and the emergence of a hypervirulent C. difficile strain. The epidemic strain NAP1 / BI / ribotype 027 is more resistant to antibiotics and produces more toxins and causes outbreaks around the world with increasing mortality and severity. Recent reports have documented that C. difficile infections occur among patients without traditional risk factors and the incidence of community-acquired CDI has increased. The clinical presentation of CDI ranging from the asymptomatic carrier to pseudomembranous colitis and toxic megacolon. The rate of recurrent CDI is 20-30 %. Early surgical consultation should be considered in all patients with severe or fulminant CDI. The preferred method for diagnosis CDI is stool testing using enzyme immunoassay (EIA) screening to look for glutamate dehydrogenase (GDH) antigen and toxins A and B. Metronidazole and vancomycin are recommended antibiotics for the treatment of CDI. Novel approaches include fidaxomicin therapy, monoclonal antibodies, and fecal microbiota transplantation. Fecal microbiota transplantation has shown relevant efficacy to overcome C. difficile infection and reduce it.

Wider bridges for roads: U. S. Department of Agriculture Press Service No. 193

Epidemiology of Clostridium difficile is characterized by worldwide increase of C. difficile infections (CDI) and the emergence of new epidemic outbreak strains with the capacity for global spreading. Long-term local surveillance at the University of Saarland Medical Center between 2000 and 2013 shows that the incidence rate of laboratory-confirmed CDI was influenced by local epidemiology as well as by testing strategies. Since 2008, molecular typing of C. difficile was regularly performed for symptomatic hospitalized patients by surface-layer protein A sequence typing (slpAST), which is an established highly standardized technique for genotyping of C. difficile. The results were assigned to known ribotypes for better comparison to international data. It could be demonstrated that distribution of genotypes was different between age groups. Older patients were predominantly infected with ribotype 001 and 027, whereas ribotype 027 was not detected in the pediatric population. Molecular typing of German isolates sent to the advisory laboratory between 2011 and 2013 revealed that ribotype 027 is present with high percentages in most German regions except for the very North.In conclusion, optimized testing of all hospitalized patients with diarrhea should be generally implemented to avoid under-diagnosis of C. difficile infection. Ribotype 027 is highly prevalent in Germany, but its infections are restricted to older patients, while absent in children. Molecular typing of suspected hospital outbreaks and of patients with severe or recurrent disease may help to better understand virulence and epidemic spreading of C. difficile.