Phage Typing Patterns Research Articles

Emerging Phage Resistance in Pseudomonas aeruginosa PAO1 Is Accompanied by an Enhanced Heterogeneity and Reduced Virulence.

Bacterial surface structures of a proteinic nature and glycoconjugates contribute to biofilm formation and provide shields to host defense mechanisms (e.g., the complement system and phagocytosis). A loss or alteration of these molecules, leading to phage resistance, could result in fewer virulent bacteria. In this study, we evaluate the biology and phenotype changes in Pseudomonas aeruginosa PAO1 phage-resistant clones, which emerge in phage-treated biofilms. We characterize these clones for phage-typing patterns, antibiotic resistance, biofilm formation, pathogenicity, and interactions with the innate immune system. Another important question that we address is whether phage-resistant mutants are also generated incidentally, despite the phage treatment-selective pressure, as the natural adaptation of the living biofilm population. It is found that the application of different phages targeting a particular receptor selects similar phage resistance patterns. Nevertheless, this results in a dramatic increase in the population heterogeneity, giving over a dozen phage-typing patterns, compared to one of the untreated PAO1 sessile forms. We also confirm the hypothesis that “phage-resistant bacteria are more susceptible to antibiotics and host-clearance mechanisms by the immune system”. These findings support phage application in therapy, although the overall statement that phage treatment selects the less virulent bacterial population should be further verified using a bigger collection of clinical strains.

Fluoroquinolone Therapy in Staphylococcus aureus Infections: Where Do We Stand?

Aim:The study aimed to evaluate the utility of various commonly used fluoroquinolones against Staphylococcus aureus isolates.Materials and Methods:A total of 250 isolates of S. aureus were studied from different clinical specimens like blood, pus, wound swabs, sputum, ear swabs, and body fluids between November 2009 and December 2011. All the isolates were tested for their susceptibility to fluoroquinolones and other antimicrobial agents by Kirby-Bauer disc diffusion method using criteria of standard zone of inhibition. Methicillin-resistant S. aureus (MRSA) detection was done by cefoxitin disk diffusion method. The MRSA isolates were tested for minimum inhibitory concentration (MIC) to vancomycin by E-test strips. All the MRSA strains were sent to National Staphylococcal Phage-typing Centre, Maulana Azad Medical College, New Delhi for phage typing.Results:A total of 107 strains of S. aureus (42.8%) were detected as MRSA. Multidrug resistance was observed among the MRSA strains more commonly than among the MSSA stains. Among the fluoroquinolones, maximum resistance in MRSA was seen to ciprofloxacin (92.5%), followed by ofloxacin (80.4%). None of the S. aureus isolates showed resistance to vancomycin and linezolid. The MICs of vancomycin for the MRSA tested ranged from 0.5 to 2 μg/ml. Phage typing pattern of 107 MRSA isolates revealed that 37 (34.6%) MRSA isolates were nontypeable and 70 (65.4%) were typeable.Conclusion:Ciprofloxacin can no longer be used in empirical therapy against MRSA infections. Use of other members of fluoroquinolone should be limited only to those strains that show laboratory confirmation of their susceptibility. Vancomycin remains the drug of choice to treat MRSA infections.

Prevalence and Characteristics of <i>Salmonella</i> spp. Isolated from Poultry Slaughterhouses in Korea

Poultry products have consistently been identified as important sources of Salmonella infection in humans, because vertical transfer of infection from breeding hens to progeny is an important aspect of the epidemiology of Salmonella spp. infection within the poultry industry. The aim of this study was to estimate the prevalence of Salmonella contamination in poultry products from 15 different located geographical areas from among the 50 poultry slaughterhouses authorized to operate in Korea and to characterize all the isolates by genotyping, phage typing and antibiotic resistance pattern. Salmonella was isolated from 10 (66.7%) of the first and 5 (33.3%) of the last chilling waters and from 32 (42.7%) carcasses originating from 9 slaughterhouses. The major prevalent serotypes of Salmonella originating from 2 duck slaughterhouses and 13 chicken slaughterhouses tested were S. Typhimurium and S. Enteritidis, respectively. Regarding the characteristics of their antibiotic resistance, 8 of the 11 ampicillin resistant (AmR) isolates carried blaTEM only, two carried blaTEM and blaCTX-M-14 and one carried blaCTX-M-3 and only one AmR isolate with the blaCTX-M-3 β-lactamase gene was an ESBL-producing Salmonella strain. Twenty-seven Salmonella isolates showed nalidixic acid resistance with a mutation at amino acid codon Asp87 in gyrA and no mutation in the parC gene. In all the phenotypic and genotypic properties of the 18 S. Enteritidis and 8 S. Typhimurium based on PFGE, phage types and antibiotic resistance pattern, the predominant patterns were XEI/BEI-PT32a-NaR (n = 5) and XTI/BTI-RNDC-no resistant antibiotics (n = 6), respectively.

Emergence of multiple drug resistance Vibrio cholerae O1 in East Delhi

Considering the changing geographical and temporal occurrence of Vibrio cholerae, there is a continuing need to monitor the strain characteristics and antibiotic resistance patterns of this pathogen. The present study was conducted to document the changing biology of V. cholerae isolates in and around Delhi, India, and the development of antibiotic resistance. A total of 1,424 stool samples or rectal swabs from patients with acute secretory diarrhoea admitted to Guru Teg Bahadur Hospital, Delhi, between January 2007 and December 2009 were processed using standard bacteriological methods. Strains identified as V. cholerae were further subjected to serogrouping, phage typing and antimicrobial susceptibility testing. Minimum inhibitory concentration (MIC) of gentamicin and tetracycline was determined. V. cholerae was isolated in 242/1,424 (17.0%) specimens. Of these, the majority were V. cholerae O1 serotype (98.3%) and serovar Ogawa. The drugs to which V. cholerae O1 isolates showed high levels of resistance were nalidixic acid, furazolidone, and cotrimoxazole throughout the study period, whereas strains were usually susceptible to chloramphenicol and cefotaxime. In 2007, there was a sudden increase of resistance to gentamicin and tetracycline, followed by a slow reversal to previous levels in subsequent years. The phage typing pattern (Basu and Mukherjee scheme) showed a dominance of phage type 2 throughout the study period. The importance of reporting all cases of V. cholerae, should be greatly emphasized, with the ultimate goal of understanding the constantly changing resistance patterns of this pathogen.

The role of transport, lairage and slaughter processes in the dissemination of Salmonella spp. in pigs in Ireland

Salmonella is an important foodborne pathogen worldwide and is commonly isolated from pigs and pig products in Ireland. Pigs, reared in an environment free of Salmonella spp. or with low levels of infection, may acquire infection or become contaminated during transport, lairage or post-slaughter. The main objective of this study was to determine the role of the abattoir as a potential factor that contributes to the dissemination of Salmonella spp. in slaughter pigs from herds with a low Salmonella seroprevalence (≤10%). A total of 128 pigs from eight herds were monitored from farm through the slaughter process in three separate abattoirs. The prevalence of Salmonella spp. was determined in samples collected from trucks, lairage pens and the slaughterline before pigs entered, from pigs after slaughter (caecal contents and ileocaecal lymph nodes) and carcass surfaces post-evisceration. Isolates were characterised by serotype, phage type and pulsed-field gel electrophoresis (PFGE) patterns. Of the swabs taken from the trucks, lairage and slaughterline, before the pigs entered, 4.3% (3/70), 80% (64/80) and 16.7% (4/27) were positive for Salmonella spp., respectively. The proportion of pigs showing serological evidence of infection was 3.1% (4/128). Salmonella spp. were isolated from the ileocaecal lymph nodes and caecal contents of 14.8% (19/128) and 11.7% (15/128) of pigs, respectively, and 13/128 (10.2%), 5/128 (3.9%), 2/111 (1.8%) and 8/111 (7.2%) carcass swabs pre wash, post wash, post chill and belly-strip samples, respectively, were Salmonella-positive. There was only slight agreement between serological and bacteriological data at the pig level. Salmonella isolates from 45% of all positive pig samples and 82% of positive carcass samples were indistinguishable, based on PFGE patterns, from salmonellae isolated from the lairage and slaughterline. Based on these results it is concluded that the lairage and the slaughterline provide a substantial source for Salmonella contamination of pigs and carcasses.

An outbreak ofSalmonellaTyphimurium DT191a associated with reptile feeder mice

In December 2008 an increase of tetracycline-resistant Salmonella Typhimurium definitive phage-type 191a (DT191a) was identified in England and Wales by the reference laboratory. This was confirmed to have a phage-typing pattern that had not previously been seen. Strong statistical evidence for an association between illness and keeping reptiles was demonstrated by a matched case-case study (mOR 16·82, 95% CI 2·78-∞). Questionnaires revealed an association with frozen reptile feeder mice, and mice representing 80% of the UK supply lines were tested for the presence of Salmonella. DT191a was found in three pools of sampled mice, which were traced back to a single supplier in the USA. Imports from this supplier were halted, and tighter regulations are now in place. A leaflet detailing how to prevent contracting Salmonella from pet reptiles has been published as well as updated advice on the Health Protection Agency's website.

Characteristics of Persistent Salmonella Enteritidis Strains in Two Integrated Broiler Chicken Operations of Korea

The objectives of the study were to investigate the phenotypic and genotypic characterization of the persistent Salmonella Enteritidis (S. Enteritidis) isolates in two integrated broiler chicken operations, with attention focused mainly on the epidemiological approach. In the distribution of virulence genes, Salmonella enterotoxin (stn), invading host cell (invA), and Salmonella plasmid virulence (spvC) genes were widely distributed among the S. Enteritidis irrespective of their source of isolation, and Salmonella fimbrial (sefC) and plasmid encoded fimbrial (pef) genes were present in 28 and 20 S. Enteritidis strains, respectively. A total of 5 different XbaI-PFGE types were obtained from 31 S. Enteritidis isolates. Twenty-one types were divided on the basis their PFGE pattern, phage type and antimicrobial resistance pattern determined. There was a significant difference in phenotypic and genotypic characterization by two integrated broiler operations. Also, 8 isolates shown susceptible to all antimicrobials and 11 isolates with resistance to nalidixic acid were partly classified by XbaI PFGE pattern and by the phage type.

Variation in Salmonella Resistance to Poultry Chemical Decontaminants, Based on Serotype, Phage Type, and Antibiotic Resistance Patterns

Chemical decontaminants are currently under review for final approval by the European Union authorities with the aim of reducing the number and/or prevalence of pathogenic microorganisms on poultry. The purpose of the research being reported here was to determine the association, if any, of decontaminant resistance with the serotype, phage type, and antibiotic resistance of Salmonella strains. Sixty poultry isolates of Salmonella enterica (serotypes Enteritidis: phage types 1, 4, 4b, 6a, 14b, and 35; Typhimurium; Newport; Infantis; Poona; Virchow; Agona; Derby; and Paratyphi B) showing resistance to none (sensitive), one (resistant), two, three, four, five, six, seven, or nine (multiresistant) antibiotics were screened for resistance to 1,000 ppm acidified sodium chlorite, 1.2% trisodium phosphate, or 25% citric acid. D-values (seconds required for 1-log reduction in the number of bacteria) in peptone water, using a linear regression, of Salmonella in the presence of acidified sodium chlorite varied widely with serotype (the highest resistance levels were shown by serotypes Typhimurium, Newport, and Derby) and antibiotic resistance pattern (average values of 8.37±1.69 s for multiresistant strains as compared with 5.96±0.54 s for sensitive, P < 0.05). A positive relationship (0.775, P < 0.001) was found between acidified sodium chlorite D-values and the number of antibiotics to which strains were resistant. Both serotype and antibiotic resistance had only a slight influence over Salmonella resistance to trisodium phosphate, with average D-values from 12.44±0.91 s (sensitive strains) to 13.28±0.77 s (multiresistant) (P < 0.05). Neither serotype nor antibiotic profile was associated with Salmonella resistance to citric acid (average D-value of 12.20±0.81 s). Minimal differences in resistance to decontaminants were found among Salmonella Enteritidis phage types. Results in the present study highlight the importance of selecting an adequate strain (serotype and antibiotic resistance pattern) when acidified sodium chlorite and trisodium phosphate are tested against Salmonella to ensure that concentrations capable of inactivating all strains are used.

Prophage Sequences Defining Hot Spots of Genome Variation inSalmonella entericaSerovar Typhimurium Can Be Used To Discriminate between Field Isolates

Sixty-one Salmonella enterica serovar Typhimurium isolates of animal and human origin, matched by phage type, antimicrobial resistance pattern, and place of isolation, were analyzed by microbiological and molecular techniques, including pulsed-field gel electrophoresis (PFGE) and plasmid profiling. PFGE identified 10 profiles that clustered by phage type and antibiotic resistance pattern with human and animal isolates distributed among different PFGE profiles. Genomic DNA was purified from 23 representative strains and hybridized to the composite Salmonella DNA microarray, and specific genomic regions that exhibited significant variation between isolates were identified. Bioinformatic analysis showed that variable regions of DNA were associated with prophage-like elements. Subsequently, simple multiplex PCR assays were designed on the basis of these variable regions that could be used to discriminate between S. enterica serovar Typhimurium isolates from the same geographical region. These multiplex PCR assays, based on prophage-like elements and Salmonella genomic island 1, provide a simple method for identifying new variants of S. enterica serovar Typhimurium in the field.

Distribution, genetic richness and phage sensitivity of Vibrio spp. from coastal British Columbia

This study examined the distribution, susceptibility to viral infection and genetic diversity of Vibrio spp. in the coastal waters and sediments of British Columbia during summer (July and August). Abundances of presumptive Vibrio spp. ranged from 1.5 to 346 ml(-1) within the water column (1-291 m); whereas, abundances at the water-sediment interface were much higher (up to approximately 3 x 10(4)Vibrio spp. cc(-1)), and decreased with sediment depth (down to 30 cm). The genetic diversity of Vibrio spp. isolates was not tied to the location from which they originated and was only influenced in a minor way by the type of environment. However, the environment had a greater effect on phage-typing patterns. Vibrio parahaemolyticus isolates from environments with high abundances of cells (sediments and oysters) were generally more susceptible to viral infection than those from the water column which were highly resistant. Therefore, although Vibrio spp. were widespread in the areas investigated, the results show that there is segregation of bacterial host strains in different environments, under differing selection pressures, which ultimately will affect in situ phage production.

Characterization of &lt;i&gt;Salmonella&lt;/i&gt; spp. Isolated from an Integrated Broiler Chicken Operation in Korea

The purpose of this study was to investigate the biological and genetic characterization of persistent Salmonella isolates in an integrated broiler chicken operation, in an attempt to elucidate the source of contamination. From the breeder farm, the hatchery, the broiler farm and the chicken slaughter house of an integrated broiler chicken operation, a total of 6 serotypes were observed. Although S. Heidelberg was not detected in the broiler farm, it was consistently found in the breeder farm, the hatchery and the chicken slaughter house. Also, S. Enteritidis and S. Senftenberg were found in the hatchery and the chicken slaughter house, and the hatchery and the broiler farm, respectively. S. Gallinarum and S. Blockley were found only in the broiler farm, and S. Virchow was only recovered in the chicken slaughter house. Isolated S. Heidelberg, S. Enteritidis and S. Senftenberg strains were divided into 3, 5 and 7 types, respectively, on the basis of all properties. Especially, S. Senftenberg isolates, divided into four types by their antimicrobial resistance patterns, were all obviously the XbaI PFGE pattern. Also, four S. Enteritidis isolates resistant to nalidixic acid showed a difference in phage type and PFGE pattern. Such a different pattern was shown despite Salmonella isolates originating from an integrated broiler operation, suggesting that further epidemiological studies on many integrated chicken companies in Korea are needed.

Shigella sonneiOutbreak among Homosexual Men, London

<i>Shigella sonnei</i>Outbreak among Homosexual Men, London

Phenotypic and genotypic differentiation of porcine Salmonella enterica subsp. enterica serovar Derby isolates

Sixty-two Salmonella enterica subsp. enterica serovar Derby isolates from slaughter pigs and meat products isolated in Southern Brazil were analyzed for their genomic relationships and for the presence of antimicrobial resistance genes. Twenty-four S. Derby isolates were indistinguishable by their subtracted restriction fingerprinting (SRF) pattern, XbaI- and BlnI-macrorestriction patterns, phage type, plasmid profile, and resistance pattern. In contrast to the BlnI-macrorestriction patterns, the XbaI-macrorestriction patterns were in good agreement with the results of SRF analysis and phage typing. Among the four phage types detected, PT10 and PT21 were the most common. The combination of all typing methods revealed a great diversity among the S. Derby isolates. All strains carried plasmids and the 60 resistant isolates showed at least tetracycline resistance. The resistance genes found were sul1 and/or sul2 (sulfonamide resistance), aadA2 (streptomycin/spectinomycin resistance), tet(A) (tetracycline resistance), tet(B) (tetracycline/minocycline resistance), bla TEM (ampicillin resistance), and dfrA14 (trimethoprim resistance). A correlation of the geno- and phenotypic characteristics with the origin of the isolates revealed a substantial temporal variation in the occurrence of specific S. Derby isolates in different independent pig production lines in Southern Brazil. The large number of resistant isolates underlined the potential risk that S. Derby isolates can pose to human health when they enter the food chain.

Antimicrobial susceptibility, phage types, and pulsetypes of Salmonella Typhimurium, in São Paulo, Brazil

A total of 283 Salmonella Typhimurium strains isolated from cases of human infections and non human sources, were examined for antimicrobial susceptibility and the incidence of resistance was 38% and multiple resistance (to three or more antimicrobials) was 15%. All 43 multidrug-resistant strains (MDR) and 13 susceptible ones were characterized by phage typing and pulsed- field gel electrophoresis (PFGE). The strains encompassed 14 definitive phage types (DT), three were untypable (UT), and 18 atypicals or reaction does not conform (RDNC), which belonged to 21 PFGE patterns, A1-A21. The predominant phage types were DT49, DT193, and RDNC and two strains belonging to DT 104 and 104b were also identified. The most common PFGE patterns were A1 and A8. Analysis by PFGE and phage typing demonstrated that the most of the MDR were multiclonal and association among multiresistance, phage typing, and PFGE patterns was not so significant.

Pulsed-field gel electrophoresis, plasmid profiles and phage types for the human isolates of Salmonella enterica serovar Enteritidis obtained over 13 years in Taiwan

Plasmid profile, phage typing, and pulsed-field gel electrophoresis (PFGE) patterns of 124 Salmonella Enteritidis strains isolated in 1998-2002 in Taiwan were analysed and the results were compared with those of the 63 strains obtained in 1991-1997, so that molecular subtypes and epidemic strains for Salmonella Enteritidis over a 13-year period (1991-2002) could be elucidated. A total of 124 strains of Salmonella Enteritidis isolated from human in Taiwan between 1998 and 2002 were analysed by PFGE, plasmid analysis and phage typing. The results obtained were compared with those of the 63 strains obtained in 1991-1997, so that the clonal relationships for a total of 187 strains obtained over 13 years could be elucidated. For PFGE, restriction enzymes XbaI, SpeI and NotI were used for chromosomal DNA digestion. Results showed 28 PFGE pattern combinations for the 187 Salmonella strains. Of them, pattern X3S3N3 was the major subtype as 130 strains isolated from different locations during 1991-2002 showed this PFGE pattern. For all these 187 strains, the genetic similarity was higher than 80%. Plasmid analysis showed 17 distinct types, which consist of one to four plasmids and the predominant phage type of those strains was PT4 (71.6%) and PT6a (13.4%). The three methods identified different degrees of polymorphism in the following order: plasmid profile (18 types, D = 0.659) > PFGE (28 types, D = 0.512) > phage typing (13 types, D = 0.438). As PFGE patterns, phage type and plasmid profile were combined for subtyping, the 187 strains could be grouped into 46 subtypes and the discriminatory index was raised to 0.795. For these 46 subtypes, the predominant one was X3S3N3/P1/PT4, which contained 77 (41%) isolates. Most of the Salmonella Enteritidis strains from sporadic cases were with pattern X3S3N3. They were the prevalent and may be the epidemic strains found in Taiwan during 1991-2002. The present study suggested that the several variants were derived from a single clonal line and the genome for strains of Salmonella Enteritidis are highly conserved over a 13-year period (1991-2002). The results obtained here are useful for epidemiolgical study of salmonellosis caused by Salmonella Enteritidis in Taiwan. Comparing the data of the present study with those obtained for strains from other countries, the major subtypes for Salmonella Enteritidis infection in the world can be elucidated.

A foodborne outbreak of Salmonella infection due to overproduction of egg-containing foods for a festival

A large outbreak of gastroenteritis occurred in Catalonia in June 2002 with 1435 cases and 117 hospitalizations. Consumption of a hard pastry with vanilla cream was strongly associated with illness. Stool samples from cases and food-handlers were analysed. The premises of the food manufacturer were inspected and food samples were taken for microbiological analysis. Salmonella serotype Enteriditis was isolated from 154 cases, three food-handlers and nine food samples. Outbreak-associated strains showed a coincident phage type, antibiotype and pulse-field gel electrophoresis pattern. Inadequate handling of foods containing eggs occurred because the establishment exceeded its safe food production capacity to meet demand for the pastry, which was consumed on the day of a traditional festival. Excessive production of foods for holidays or special events represents a potential public health threat.

Asymptomatic carriage of verocytotoxin-producing Escherichia coli O157 in farm workers in Northern Italy.

Faecal samples from 350 farm workers on 276 dairy farms and 50 abattoir employees from seven different operations were examined for the presence of Verocytotoxin-producing Escherichia coli 0157 (VTEC O157) by an O157-specific enzyme-linked fluorescent assay followed by immunoconcentration. VTEC O157 was isolated from four (1.1%) of the farm workers. A second stool sample was obtained from the positive farm workers as well as from their household contacts. VTEC O157 was isolated from the wife of one of them. The strains from the same household shared the same Verocytotoxin genes profile, phage type and pulsed-field gel electrophoresis pattern. The VTEC O157-positive subjects had neither intestinal symptoms at the moment of sampling nor a history of bloody diarrhoea or renal failure. Our study seems to confirm the hypothesis that farm residents often develop immunity to VTEC O157 infection, possibly due to recurrent exposure to less virulent strains of VTEC.

Diversity of phage types among archived cultures of the Demerec collection of Salmonella enterica serovar Typhimurium strains.

The existence of several thousand Salmonella enterica serovar Typhimurium LT2 and LT7 cultures originally collected by M. Demerec and sealed in agar stab vials for 33 to 46 years is a resource for evolutionary and mutational studies. Cultures from 74 of these vials, descendants of cells sealed and stored in nutrient agar stabs several decades ago, were phage typed by the Callow and Felix, Lilleengen, and Anderson systems. Among 53 LT2 archived strains, 16 had the same phage type as the nonarchival sequenced LT2 strain. The other 37 archived cultures differed in phage typing pattern from the sequenced strain. These 37 strains were divided into 10 different phage types. Among the 19 LT7 strains, only one was similar to the parent by phage typing, while 18 were different. These 18 strains fell into eight different phage types. The typing systems were developed to track epidemics from source to consumer, as well as geographic spread. The value of phage typing is dependent upon the stability of the phage type of any given strain throughout the course of the investigation. Thus, the variation over time observed in these archived cultures is particularly surprising. Possible mechanisms for such striking diversity may include loss of prophages, prophage mosaics as a result of recombination events, changes in phage receptor sites on the bacterial cell surface, or mutations in restriction-modification systems.

Alterations in phage-typing patterns in vancomycin-intermediate Staphylococcus aureus.

The ability of phage-typing and SmaI chromosomal RFLPs to conclude appropriate strain relatedness between a collection of 12 well-characterized in vitro-selected vancomycin-intermediate Staphylococcus aureus (VISA) strains and their seven vancomycin-susceptible parent strains is reported. Generally, no SmaI RFLP alterations were observed in VISA strains when they were compared with their respective parent strains, and clonal relationships between isogenic strains were clearly evident. Unlike the SmaI RFLP results, parent strains and VISA derivatives generally did not share similar phage-typing profiles. Depending on the phage set investigated, some VISA strains even became untypable by this method. Loss of phage infectivity is probably due to cell wall (phage receptor) alterations that are expressed by the VISA strains investigated. Collectively, these findings indicate that inappropriate relationships between VISA and vancomycin-susceptible parents might be drawn if only phage-typing and antibiotic susceptibility are utilized to determine epidemiological relationships.

The prevalence and spread of Escherichia coli O157:H7 at a commercial beef abattoir.

To investigate the prevalence and virulence characteristics of Escherichia coli O157:H7 after a number of beef process operations at a commercial Irish abattoir. Two 12-month studies were carried out. The first study (study 1) examined the prevalence of E. coli O157:H7 at up to six sites on carcasses at eight stages of the dressing, washing, chilling and boning process. The second study (study 2) examined the prevalence of E. coli O157:H7 in bovine faeces and rumen contents post-slaughter and on dressed, washed carcasses. Isolates from both studies were phage-typed and the presence of genes encoding verocytotoxin, enterohaemolysin and intimin production was determined. E. coli O157:H7 was isolated from four of 36 carcasses in study 1. E. coli O157:H7 was detected during hide removal and was detected at multiple carcass sites and multiple process stages, including boning. On two carcasses, contamination was first detected at the bung following its freeing and tying. All isolates from study 1 were phage type (PT) 2, eaeAO157 and ehlyA positive, but were verocytotoxin 1 (VT1) and verocytotoxin 2 (VT2) negative. In study 2, E. coli O157:H7 was isolated from 2.4% of faecal, 0.8% of rumen and 3.2% of carcass samples. In some cases, isolates recovered from the faeces of a particular animal, the resulting carcass and adjacent carcasses on the line had the same phage typing and virulence characteristic profile patterns. All isolates from study 2 were eaeAO157 and ehlyA positive and only one isolate was VT1 and VT2 negative. Most isolates were PT 32. A higher frequency of positive isolations was noted from samples taken during spring and late summer. These studies show that in a typical Irish beef abattoir, carcass contamination with E. coli O157:H7 can occur during hide removal and bung tying and this contamination can remain on the carcass during subsequent processing. This study provides data that is necessary for the understanding of how E. coli O157:H7 contamination of beef occurs.