Dynamics of Campylobacter colonization of a natural host, Sturnus vulgaris (European starling)

Wild European Starlings ( Sturnus vulgaris ) shed Campylobacter at high rates, suggesting that they may be a source of human and farm animal infection. A survey of Campylobacter shedding of 957 wild starlings was undertaken by culture of faecal specimens and genetic analysis of the campylobacters isolated: shedding rates were 30.6% for Campylobacter jejuni , 0.6% for C. coli and 6.3% for C. lari. Genotyping by multilocus sequence typing (MLST) and antigen sequence typing established that these bacteria were distinct from poultry or human disease isolates with the ST-177 and ST-682 clonal complexes possibly representing starling-adapted genotypes. There was seasonal variation in both shedding rate and genotypic diversity, both exhibiting a maximum during the late spring/early summer. Host age also affected Campylobacter shedding, which was higher in younger birds, and turnover was rapid with no evidence of cross-immunity among Campylobacter species or genotypes. In nestlings, C. jejuni shedding was evident from 9 days of age but siblings were not readily co-infected. The dynamics of Campylobacter infection of starlings differed from that observed in commercial poultry and consequently there was no evidence that wild starlings represent a major source of Campylobacter infections of food animals or humans.     

Prevalence of Campylobacter spp. in a subset of intensive poultry flocks in Ireland

Aim:  The aim of this study was to investigate the prevalence of Campylobacter species in a subset of intensive poultry flocks by examining samples collected in geographically disparate areas on the island of Ireland.
Methods and Results:  Faecal, water and environmental samples were collected from the interior of poultry houses on nine farms. Three cultural methods were used for Campylobacter isolation: direct plating, enrichment culture and a recovery method for emerging Campylobacter spp. Presumptive Campylobacter isolates were confirmed using biochemical tests and further identified to species level by multiplex PCR. All flocks sampled in this study were found to be contaminated with Campylobacter at the time of sampling. Structural and air samples taken from the interior of broiler houses were also found to be Campylobacter positive. All water samples were found to be Campylobacter negative. The Campycheck method was used for the isolation of emerging Campylobacter spp.
Conclusions:  Campylobacter spp. were recovered (as contaminants) from the poultry house interior, air and environmental samples in all intensive poultry flocks surveyed.
Significance and Impact of the Study:  This study highlights the need for improved biosecurity on selected poultry farms.     

Genetic instability is associated with changes in the colonization potential of Campylobacter jejuni in the avian intestine

Aims:  A panel of pulsed field gel electrophoresis (PFGE) type variants of Campylobacter jejuni , previously identified as of clonal origin, were investigated to determine whether genomic instability could be observed during competitive growth.
Methods and Results:  Upon recovery from frozen storage, some variants had undergone alterations in PFGE profiles, but subsequent culture produced constant genotypes. Individual variants did not display differences in colonization potential when tested in orally challenged 1-day-old chickens. However, competitive colonization using mixtures of two or three PFGE types generally resulted, by 4 weeks postchallenge, in one predominant PFGE type in all birds. For some variant mixtures, a minor population of novel PFGE types was detected in individual birds. The creation of new variants appeared to be dependent on the extent of competition and of the individual host. Genomic rearrangements most likely explain this increase in genetic diversity, apparently without the involvement of natural transformation or plasmid acquisition. In vitro cultivation of mixed inoculations were again selected for particular variants; but genetic diversity was not generated, suggesting that the selection pressures in vitro differed from those active in vivo .
Conclusion:  These observations support the hypothesis that by generating genetic diversity, C. jejuni can improve its phenotypic fitness to survive and colonize subsequent hosts.
Significance and Impact of the Study:  The consequences of such observations for the development of campylobacter control strategies for poultry may be substantial.     

Characterization of vanA-type Enterococcus faecium isolates from urban and hospital wastewater and pigs

Aims:  In this study we analysed urban, hospital wastewater and pig faeces samples to investigate the presence of vancomycin-resistant Enterococcus faecium strains (VREF) and to determine potential links among the strains originating from the above sources and VREF strains causing clinical infections.
Methods and Results:  Urban, hospital wastewater and pig faeces exhibited high VREF prevalence of 52%, 87% and 85%, respectively. Pulsed field gel electrophoresis (PFGE) clustering of VREF genotypes as well as discriminant analysis of antibiotic resistance patterns of VREF strains revealed their source specificity while strains isolated from hospitalized humans were genetically distinct.
Conclusions:  PFGE genotypes and antimicrobial resistance patterns in VREF isolates are distinguishable by each sample origin. The observed high genetic diversity of VREF suggests horizontal transfer of genetic elements among VREF. Phenotypic and genotypic data indicate that VREF isolates of hospital-treated wastewater might pass to the urban wastewater system.
Significance and Impact of the Study:  This study provides information to understand the origin and the mechanism of circulation of vancomycin resistance in food animals and wastewater treatment plants for minimizing the risk of transmission of VRE in human population.     

Delineation of Pseudomonas savastanoi pv. savastanoi strains isolated in Tunisia by random-amplified polymorphic DNA analysis

Aims:  To investigate the genetic diversity of Pseudomonas savastanoi pv. savastanoi strains and to look whether these strains were distributed to geographical location.
Methods and Results:  Random amplification of polymorphic DNA (RAPD) was used to discriminate between 58 Tunisian strains and 21 strains from various other countries of P. savastanoi pv. savastanoi , the causal agent of olive knot disease. Isolates were separated into three groups by cluster analysis and principal coordinate analysis of RAPD fingerprint data obtained with three primers (OPR-12, OPX-7 and OPX-14). Group 1 contained isolates from the southeast of Tunisia and European strains. Group 2 comprised strains isolated from the north of Tunisia exclusively while group 3 encompassed the majority of isolates obtained from five orchards located in the centre of Tunisia.
Conclusions:  The results indicated that isolates of P. savastanoi pv. savastanoi were genetically distinct according to geographic regions. RAPD grouped isolates derived from the same orchard as identical.
Significance and Impact of the Study:  This is the first application of RAPD in the delineation of P. savastanoi pv. savastanoi strains.     

A genotyping investigation of the colonization of piglets by Campylobacter coli in the first 10 weeks of life

Aims:  To examine the Campylobacter genotypes colonizing a litter of piglets during the first 10 weeks of life and compare them with those of the sow.
Methods and Results:  Campylobacters were isolated by direct plating of anal swabs. Piglets ( n  = 6) were sampled six times and five isolates per piglet obtained each time. The sow was also sampled but 20 isolates per sampling obtained. Isolates were genotyped by random amplification of polymorphic DNA, pulsed field gel electrophoresis and polymerase chain reaction/restriction fragment length polymorphism of the flagellin gene.
Initially piglets were colonized by Campylobacter coli genotypes from the mother but after 66 days 33% of piglet isolates (based on a mean of the three-genotyping methods) were from other sources. The sow died after 14 days and the initial Campylobacter flora of the foster sow was subsequently replaced by genotypes from the piglets mother. However these constituted only a minor part of her flora after 52 days. Both foster sow and piglets carried multiple genotypes of Camp. coli : up to four in a single piglet sample and seven in one from the sow.
Significance and Impact of the Study:  Piglets are initially colonized by Camp. coli genotypes from their mother but later other genotypes displace them. Colonization is dynamic with the sow able to acquire genotypes from the piglets.
Conclusions:  The large numbers of Camp. coli genotypes carried by pigs, and frequent successions of dominant types, will render epidemiological studies problematic.     

Molecular subtype analyses of Campylobacter spp. from Arkansas and California poultry operations

Campylobacter isolates from diverse samples within broiler production and processing environments were typed by using flaA short variable region DNA sequence analysis. Sixteen flocks from four different farms representing two broiler producers in Arkansas and California were analyzed. Fourteen of the flocks (87.5%) were Campylobacter-positive; two remained negative throughout the 6-week rearing period. In general, multiple clones were present within a flock. Additionally, clones found within a flock were also present on the final product, although the diversity of Campylobacter spp. on the final product appeared to be reduced relative to that observed within the flock. Comparison of clones between flocks on the same farm revealed that some clones of Campylobacter persisted in multiple flocks. Furthermore, some clones were identified across the two farms that were under the same management. In two sampling periods, environmental isolates were positive for Campylobacter prior to flock shedding. Environmental samples associated with five additional flocks were positive for Campylobacter concomitantly with recovery of Campylobacter from the birds. Analysis of the environmental isolates that were positive prior to flock shedding demonstrated that in some instances the environmental isolates possessed genotypes identical to those of isolates originating from the flock, while in other cases the environmental isolates possessed genotypes that were distantly related to isolates obtained from the flock. Analyses of environmental isolates that tested positive concurrently with the positive isolates from the flocks demonstrated varied results; in some instances the environmental isolates possessed genotypes identical to those of isolates originating from the flock, while in other cases the environmental isolates possessed genotypes that were distantly related to isolates obtained from the flock. These data suggest that the external environment may contribute to Campylobacter contamination during poultry production and processing. However, environmental contamination with Campylobacter does not appear to be the sole contributing factor.     

Genetic characterization and antibiotic resistance of Campylobacter spp. isolated from poultry and humans in Senegal

AIMS: The main objectives of this study were to investigate the diversity of Campylobacter genotypes circulating in Senegal and to determine the frequency of antibiotic resistance. METHODS AND RESULTS: Strains of Campylobacter jejuni isolated from poultry (n = 99) and from patients (n = 10) and Campylobacter coli isolated from poultry (n = 72) were subtyped by pulsed-field gel electrophoresis (PFGE). The pulsotypes obtained after digestion by SmaI and KpnI revealed a significant genetic diversity in both species, but without any predominant pulsotypes. However, farm-specific clones were identified in the majority of poultry houses (76.5%). Human and poultry isolates of C. jejuni had common PFGE patterns. High quinolone-resistance rates were observed for C. jejuni (43.4%) and C. coli (48.6%) isolates obtained from poultry. CONCLUSIONS: The results showed a genetic diversity of Campylobacter between farms indicating multiple sources of infection; but specific clones had the ability to colonize the broiler farms. The antimicrobial resistance patterns were not related to any specific PFGE pattern suggesting that resistance was due to the selective pressure of antibiotic usage. Campylobacter with similar genotypes were circulating in both human and poultry. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is important for the understanding of the epidemiology of Campylobacter in broiler farms in Senegal. It also emphasizes the need for a more stringent policy in the use of antimicrobial agents in food animals.     

Antimicrobial resistance profiling and fla-typing of Irish thermophillic Campylobacter spp. of human and poultry origin

AIMS: To determine the genetic relationship between a random collection of Irish human and poultry Campylobacter isolates and to determine the frequency of antibiotic resistance. METHODS AND RESULTS: Sixty-six Campylobacter isolates (34 human and 32 poultry) were typed by restriction fragment length polymorphism of flagellin-A (flaA) PCR products (flaA-RFLP) and by sequencing of the short variable region of flaA (flaA-SVR). FlaA-RFLP identified 58 distinct profiles, while flaA-SVR identified 28 different alleles. The highest level of antibiotic resistance was found for ampicillin (48.5%) followed by nalidixic acid (42.2%) and ciprofloxacin (31.8%). In general, poultry isolates displayed a higher incidence of resistance to the antimicrobials tested than the human isolates. CONCLUSIONS: A high level of genetic diversity existed among the Campylobacter strains confirming the weak clonality of this species. There was no relationship between antimicrobial resistance and specific genotypes determined by flaA typing suggesting that resistance was due to pressures of antimicrobial therapy. SIGNIFICANCE AND IMPACT OF THE STUDY: This study emphasizes the need for increased surveillance of antimicrobial susceptibility among Campylobacter isolates in Ireland and the implementation of more stringent control policies relating to the use of antimicrobials in the poultry industry.     

Genomic diversity of Campylobacter coli and Campylobacter jejuni isolates recovered from free-range broiler farms and comparison with isolates of various origins

In many industrialized countries, the incidence of campylobacteriosis exceeds that of salmonellosis. Campylobacter bacteria are transmitted to humans mainly in food, especially poultry meat products. Total prevention of Campylobacter colonization in broiler flocks is the best way to reduce (or eliminate) the contamination of poultry products. The aim of this study was to establish the sources and routes of contamination of broilers at the farm level. Molecular typing methods (DNA macrorestriction pulsed-field gel electrophoresis and analysis of gene polymorphism by PCR-restriction fragment length polymorphism) were used to characterize isolates collected from seven broiler farms. The relative genomic diversity of Campylobacter coli and Campylobacter jejuni was determined. Analysis of the similarity among 116 defined genotypes was used to determine clusters within the two species. Furthermore, evidence of recombination suggested that there were genomic rearrangements within the Campylobacter populations. Recovery of related clusters from different broiler farms showed that some Campylobacter strains might be specifically adapted to poultry. Analysis of the Campylobacter cluster distribution on three broiler farms showed that soil in the area around the poultry house was a potential source of Campylobacter contamination. The broilers were infected by Campylobacter spp. between days 15 and 36 during rearing, and the type of contamination changed during the rearing period. A study of the effect of sanitary barriers showed that the chickens stayed Campylobacter spp. free until they had access to the open area. They were then rapidly colonized by the Campylobacter strains isolated from the soil.     

A genetically homogenous population of Fusarium circinatum causes pitch canker of Pinus radiata in the Basque Country, Spain

Pitch canker, caused by Fusarium circinatum, is a destructive disease of Pinus species and has recently been shown to represent a substantial threat to natural and commercial forests in northern Spain. The genetic diversity of F. circinatum in the Basque Country of Spain was assessed by characterising 96 isolates based on vegetative compatibility groups (VCGs), mating type assays, polymorphic DNA-markers and amplified fragment length polymorphism (AFLP) analyses. For this purpose, F. circinatum isolates were collected from diseased Pinus radiata as well as from insects associated with this host. Overall, a low level of diversity was detected in the population. The isolates represented only two VCGs and they were all of the same mating type. AFLP analyses revealed three genotypes and polymorphic DNA-markers specific for F.?circinatum showed nine genotypes. The most common genotypes represented 97% of all isolates for AFLP analysis and 68% of isolates for the polymorphic DNA-marker sets. Over all, this indicates that pitch canker in the Basque Country of Spain is caused by a clonally propagating population of F. circinatum, typical of a recently introduced pathogen.     

Genetic diversity of Campylobacter jejuni isolates from farm animals and the farm environment

The genetic diversity of Campylobacter jejuni isolates from farm animals and their environment was investigated by multilocus sequence typing (MLST). A total of 30 genotypes, defined by allelic profiles (assigned to sequence types [STs]), were found in 112 C. jejuni isolates originating in poultry, cattle, sheep, starlings, and slurry. All but two of these genotypes belonged to one of nine C. jejuni clonal complexes previously identified in isolates from human disease and retail food samples and one clonal complex previously associated with an environmental source. There was some evidence for the association of certain clonal complexes with particular farm animals: isolates belonging to the ST-45 complex predominated among poultry isolates but were absent among sheep isolates, while isolates belonging to the ST-61 and ST-42 complexes were predominant among sheep isolates but were absent from the poultry isolates. In contrast, ST-21 complex isolates were distributed among the different isolation sources. Comparison with MLST data from 91 human disease isolates showed small but significant genetic differentiation between the farm and human isolates; however, representatives of six clonal complexes were found in both samples. These data demonstrate that MLST and the clonal complex model can be used to identify and compare the genotypes of C. jejuni isolates from farm animals and the environment with those from retail food and human disease.     

Genetic variability of Campylobacter jejuni isolated from fresh and frozen broiler carcasses

AIMS: The aim of this study was to determine the genetic variability of Campylobacter jejuni isolates from poultry before and after freezing treatment in order to identify genotypes that would survive the treatment. METHODS AND RESULTS: C. jejuni was isolated from both fresh and frozen halves of the same carcass after freezing for 2 or more than 20 days at -20 degrees C. From 36 carcasses, representing five unrelated flocks in Norway, a total of 209 isolates were included in the study. Thirty-two of the isolates were recovered with a qualitative method while the remaining 177 were isolated using a quantitative method. Isolates were genotyped with fluorescent amplified fragment length polymorphism using MfeI and BglII restriction enzymes. Nine different genotypes were identified, however, one genotype was shown to be dominant in three different flocks. This genotype and the dominant genotype of another flock were found among isolates from fresh and frozen broiler halves. They were also shown to be identical to genotypes frequently identified among strains isolated from humans, cattle and poultry flocks in previous years. CONCLUSIONS: Freezing treatment or isolation method appeared not to select for a particular genotype. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of the present study indicate that the freezing tolerance of strains is not genotype dependent.     

Comparison of genotypes and antibiotic resistance of Campylobacter jejuni isolated from humans and slaughtered chickens in Switzerland

Aims: To get an overview of genotypes and antibiotic resistances in Swiss Campylobacter jejuni implicated in human gastroenteritis and to examine the association with isolates from chickens. Methods and Results: Multilocus sequence typing (MLST) and flaB typing were applied to 136 human clinical isolates. Phenotypic resistance to 12 antimicrobials and genotypic resistance to macrolides and quinolones were determined. MLST resulted in 35 known and six new sequence types (ST). The flaB analysis revealed 35 different types, which – in combination with MLST – increased the resolution of the typing approach. Resistance to quinolones, tetracycline and ampicillin was found in 37·5, 33·1 and 8·1% of the isolates, respectively, whereas macrolide resistance was found only once. Genotypic and phenotypic resistance correlated in all cases. A comparison to Camp. jejuni isolated from slaughtered chickens was performed. While 86% of the quinolone‐sensitive human isolates showed overlapping MLST‐flaB types with those of chicken origin, resistant strains showed only 39% of matching types. Conclusion: Mainly quinolone‐sensitive Camp. jejuni strains implicated in human campylobacteriosis showed matching genotypes with isolates originating from chickens. Significance and Impact of the Study: A large proportion of human cases in Switzerland are likely to originate from domestic chickens, confirming that prevention measures in the poultry production are important.     

Comparison of Lactococcus garvieae strains isolated in northern Italy from dairy products and fishes through molecular typing

Aims:  To investigate the genetic relatedness between Lactococcus garvieae strains isolated from fish and dairy samples collected in northern Italy, using random-amplified polymorphic DNA (RAPD)-polymerase chain reaction (PCR), Sau -PCR and amplified fragment length polymorphism (AFLP).
Methods and Results:  Eighty-one isolates from bovine and caprine dairy products ( n  = 53) and from diseased rainbow trouts and other fishes ( n  = 28) were examined. All methods showed a typeability of 100%, repeatability ranging from 84·4% to 97·5% and discriminatory powers from 0·798 to 0·986. Dairy and fish strains revealed a low genetic relatedness as they are often grouped into distinct clusters. RAPD analysis discriminated 52 genotypes when primer M13 was used, whereas with primer P5 only 27 genotypes were identified. When Sau -PCR was performed, 13 genotypes were detected while AFLP analysis allowed the differentiation of 32 genotypes.
Conclusion:  L. garvieae strains isolated from dairy samples are generally not related to those collected from fish lactococcosis outbreaks.
Significance and Impact of the Study:  L. garvieae strains exhibit a genetic diversity related to the specific animal host they colonize. RAPD M13 fingerprinting proved to be a molecular tool for comparing isolates, whereas Sau -PCR and AFLP analyses were useful techniques to investigate the distribution of L. garvieae populations in the environment.     

Genetic diversity in Helicobacter pullorum from human and poultry sources identified by an amplified fragment length polymorphism technique and pulsed-field gel electrophoresis

Helicobacter pullorum was first isolated from the faeces and carcasses of poultry and has been associated with human gastroenteritis. The aim of this study was to examine interstrain genetic diversity within H. pullorum. Two fingerprinting techniques were used: amplified fragment length polymorphism (AFLP) and pulsed field gel electrophoretic (PFGE) analysis. The 20 strains examined were from four countries and comprised 13 human isolates and seven poultry isolates. Their identity was confirmed by a species-specific PCR assay. The human and poultry isolates had distinct genotypes and most strains showed a high degree of genetic diversity. Genotyping also indicated a clonal origin for two strains from the same poultry flock, and established a close relatedness between three chicken carcass isolates from a processing plant. It is concluded that these two genotyping techniques will provide a useful basis for future epidemiological investigations of H. pullorum in poultry, and may provide a link with its possible causal role in human gastrointestinal infections.     

Effect of using a stand-off pad on Campylobacter jejuni strain diversity in a herd of dairy cows

Aim:  To determine the effect of stand-off pad (SOP) use on the prevalence and strain diversity of Campylobacter jejuni in a small herd of dairy cows.
Methods and Results:  Faecal samples were collected from 21 cows on four sampling occasions (events), one in each season, over 1 year. The cows usually grazed on pasture but during winter they spent 18 h a day on a SOP. Campylobacter prevalence ranged from 48–52% on pasture but was 62% on the SOP. The diversity of 386 C. jejuni isolates was determined using Enterobacterial Repetitive Intergenic Consensus polymerase chain reaction (ERIC/PCR). There were 11 ERIC types identified for the herd over the course of the study. Of those 11, four to seven (per event) were present when the cows were grazing pasture but only two during SOP use.
Conclusions:  The use of the SOP was associated with an increase in prevalence and a reduction in diversity of C. jejuni.
Significance and Impact of the Study:  The reduction in ERIC types on the SOP indicated an increase in transfer of only some strains of C. jejuni among the cows. One of these strains persisted throughout the study. The zoonotic potential of this strain warrants further investigation.     

Performance of the potentially invasive Siberian moth Dendrolimus superans sibiricus on coniferous species in Europe

1 The native range of the Siberian moth extends from the Pacific Ocean (Russian Far East, Japan and Northern Korea) across Siberia, Northern China and Mongolia to the Ural Mountains. At the beginning of the 21st Century, this species was documented west of the Ural Mountains in the Republic of Mari El, indicating range extension toward the west.
2 The Siberian moth has recently been suggested for regulation as a quarantine pest for European and Mediterranean Plant Protection Organization member countries. However, no specific report on European host plants for this pest has been published so far.
3 In the present study, larval host plant choice and performance was tested for the first time on coniferous tree species that are widely distributed and of commercial value in Europe.
4 Based on dual-choice tests on neonates and mortality, developmental duration and relative growth rates of the first- to sixth-instar larvae, we found European larch Larix decidua to be the most suitable host for the moth larvae, whereas European black pine Pinus nigra and Scots pine Pinus sylvestris were the poorest hosts. The remaining conifer species tested, European silver fir Abies alba , Nordmann fir Abies nordmanniana , and Norway spruce Picea abies , were intermediate host plants. Douglas-fir Pseudotsuga menziesii , originating from North America, was chosen by the larvae to the same extend as European larch, and was also highly suitable for larval development.
5 If the moth is introduced to European countries, it will become damaging in stands of European larch and Douglas-fir, mixed stands of fir and spruce; however, it will be less damaging in forests dominated by two-needle pines.
6 We predict that Dendrolimus superans sibiricus will be able to survive and develop on the main European coniferous tree species, including non-native coniferous tree species, resulting in severe damage to large areas of forests.     

Comparison of genotypes and serotypes of Campylobacter jejuni isolated from Danish wild mammals and birds and from broiler flocks and humans.

The incidence of human infection with Campylobacter jejuni is increasing in most developed countries and the reason for this is largely unknown. Although poultry meat is considered to be a major source, it is evident that other reservoirs exist, possibly common to humans and poultry. Environmental sources are believed to be important reservoirs of Campylobacter infection in broiler chicken flocks. We investigated the potential importance of wildlife as a source of infection in commercial poultry flocks and in humans by comparing the serotype distributions, fla types, and macrorestriction profiles (MRPs) of C. jejuni isolates from different sources. The serotype distribution in wildlife was significantly different from the known distributions in broilers and humans. Considerable sero- and genotype diversity was found within the wildlife collection, although two major groups of isolates within serotype O:12 and the O:4 complex were found. Common clonal lines among wildlife, chicken, and/or human isolates were identified within serotype O:2 and the O:4 complex. However, MRPs of O:12 and O:38 strains isolated from wildlife and other sources indicated that some clonal lines propagated in a wide selection of animal species but were not detected in humans or broilers in this study. The applied typing methods successfully identified different clonal groups within a strain collection showing large genomic diversity. However, the relatively low number of wildlife strains with an inferred clonal relationship to human and chicken strains suggests that the importance of wildlife as a reservoir of infection is limited.     

Genotypic characterization of hospital Enterococcus faecalis strains using multiple-locus variable-number tandem-repeat analysis

Aims:  The level of genetic diversity and relationships between the specific genotypes and the distribution of virulence determinants among Enterococcus faecalis strains isolated from patients hospitalized in different wards of two hospitals were investigated.
Methods and Results:  Fifty-six clinical strains of E. faecalis , isolated from patients hospitalized in the period of 1999–2004 in several wards in Wrocław (Poland), were analysed by multiple-locus variable-number tandem-repeat analysis (MLVA). Analysis of seven genomic loci identified 40 novel genotypes among the analysed E. faecalis strains, with two major genomic groups, designated I and II, distinguished at a cut-off of 35%. With a similarity cut-off of 85·7%, the genotypes could be combined into 12 clusters (C1–C12), containing at least two isolates. The remaining 18 MLVA types were represented by a single isolate.
Conclusions:  Based on the data obtained by MLVA, it was found that (i) many E. faecalis isolates recovered from patients from the wards whose location allowed the potential transmission of micro-organisms, belonged to closely related MLVA types and (ii) possible relationships between specific E. faecalis genotype and the virulence factors lipase, haemolysin and esp gene can exist.
Significance and Impact of the Study:  Our study confirms that MLVA is a suitable method for the epidemiological study of E. faecalis and for the first time shows possible relationships between specific genotypes and such virulence determinants, i.e. lipase, haemolysin and esp gene.