Effect of the growth promoter avilamycin on emergence and persistence of antimicrobial resistance in enteric bacteria in the pig

AIM: To assess the effect of the growth promoter avilamycin on emergence and persistence of resistance in enteric bacteria in the pig. METHODS AND RESULTS: Pigs (treated with avilamycin for 3 months and controls) were challenged with multi-resistant Salmonella Typhimurium DT104 and faecal counts were performed for enterococci, Escherichia coli, S. Typhimurium and Campylobacter (before, during and 5 weeks post-treatment). Representative isolates were tested for antibiotic resistance and for the presence of resistance genes. Avilamycin-resistant Enterococci faecalis (speciated by PCR) were isolated from the treated pigs and continued to be detected for the first week after treatment had ceased. The avilamycin-resistance gene was characterized by PCR as the emtA gene and speciation by PCR. MIC profiling confirmed that more than one strain of Ent. faecalis carried this gene. There was no evidence of increased antimicrobial resistance in the E. coli, Salmonella and Campylobacter populations, although there was a higher incidence of tetB positive E. coli in the treated pigs than the controls. CONCLUSION: Although avilamycin selects for resistance in the native enterococci population of the pig, no resistant isolates were detected beyond 1 week post-treatment. This suggests that resistant isolates were unable to persist once selective pressure was removed and were out-competed by the sensitive microflora. SIGNIFICANCE AND IMPACT OF THE STUDY: Our data suggest the risk of resistant isolates becoming carcass contaminants and infecting humans could be minimized by introducing a withdrawal period after using avilamycin and prior to slaughter.     

National survey for Salmonella in pigs, cattle and sheep at slaughter in Great Britain (1999-2000)

AIMS: The objective of these surveys was to estimate the prevalence of faecal carriage of Salmonella in healthy pigs, cattle and sheep at slaughter, and of pig carcase contamination with Salmonella. These data can be used as a baseline against which future change in Salmonella prevalence in these species at slaughter can be monitored. METHODS AND RESULTS: In this first randomized National Survey for faecal carriage of Salmonella in slaughter pigs, cattle and sheep in Great Britain, 2509 pigs, 891 cattle and 973 sheep were sampled in 34 pig abattoirs and 117 red meat abattoirs in England, Scotland and Wales. Carriage of Salmonella in 25 g caecal contents was identified in 578 (23.0% pigs) but in only 134 (5.3%) of carcase swabs. The predominant Salmonella serovars found in both types of sample were S. Typhimurium (11.1% caeca, 2.1% carcases) and S. Derby (6.3% caeca, 1.6% carcases). The main definitive phage types (DT) of S. Typhimurium found were DT104 (21.9% of caecal S. Typhimurium isolates), DT193 (18.7%), untypable strains (17.6%), DT208 (13.3%) and U302 (13.3%). Three isolates of S. Enteritidis (PTs 13A and 4) and one enrofloxacin-resistant S. Choleraesuis were also isolated. A positive 'meat-juice ELISA' was obtained from 15.2% of pigs at 40% optical density (O.D.) cut-off level and 35.7% at 10% cut-off. There was poor correlation between positive ELISA results or carcase contamination and the caecal carriage of Salmonella. The ratio of carcase contamination to caecal carriage rates was highest in abattoirs from the midland region of England and in smaller abattoirs. In cattle and sheep 1 g samples of rectal faeces were tested. Two isolates (i.e. 0.2%) were recovered from cattle, one each of S. Typhimurium, DT193 and DT12. One sheep sample (0.1%) contained a Salmonella, S. Typhimurium DT41. In a small subsidiary validation exercise using 25 g of rectal faeces from 174 cattle samples, three (1.7%) isolates of Salmonella (S. Typhimurium DT104, S. Agama, S. Derby) were found. CONCLUSIONS: The carriage rate of Salmonella in prime slaughter cattle and sheep in Great Britain was very low compared with pigs. This suggests that future control measures should be focused on reduction of Salmonella infection on pig farms and minimizing contamination of carcases at slaughter. SIGNIFICANCE AND IMPACT OF THE STUDY: This work has set baseline figures for Salmonella carriage in these species slaughtered for human consumption in Great Britain. These figures were collected in a representative way, which enables them to be used for monitoring trends and setting control targets.     

Adaptive responses of Salmonella enterica serovar Typhimurium DT104 and other S. Typhimurium strains and Escherichia coli O157 to low pH environments

AIMS: Cattle are a known main reservoir for acid-resistant Escherichia coli O157 and Salmonella enterica serovar Typhimurium DT104. We studied the response of S. Typhimurium DT104 to extreme low pH environments and compared their response to that of acid-resistant E. coli O157 and other S. Typhimurium phage types. METHODS AND RESULTS: Bacteria were grown in nutrient-rich medium and subsequently acid challenged at pH 2.5. We found that stationary phase cultures of various S. Typhimurium strains were able to survive a challenge for 2 h at pH 2.5. As in E. coli, the ability of S. Typhimurium to survive at pH 2.5 was shown to be dependent on the presence of amino acids, specifically arginine. The amount of proton pumping H+/ATPase, both in E. coli O157 and S. Typhimurium strains, was lower when grown at pH values <6 than after growth at pH 7.5. Cyclo fatty acid content of membranes of bacteria grown at pH values <6 was higher than that of membranes of bacteria grown at pH 7.5. CONCLUSIONS: Various S. Typhimurium strains, both DT104 and non-DT104, are able to survive for a prolonged period of time at pH 2.5. Their response to such low pH environment is seemingly similar to that of E. coli O157. SIGNIFICANCE AND IMPACT OF THE STUDY: Food-borne pathogens like S. Typhimurium DT104 and E. coli O157 form a serious threat to public health since such strains are able to survive under extreme low pH conditions as present in the human stomach. The emergence these acid-resistant strains suggests the presence of a selection barrier. The intestinal tract of ruminants fed a carbohydrate-rich diet might be such a barrier.     

The prevalence of Salmonella spp. in bovine faecal,rumen and carcass samples at a commercial abattoir

AIMS: To determine the prevalence, serotype and antibiotic resistance profile of Salmonella isolates in cattle and on carcasses at a commercial Irish abattoir. METHODS AND RESULTS: Faecal, rumen and carcass samples were collected from a beef abattoir over a 12-month period and examined for the presence of Salmonella spp. Isolates were serotyped, phage typed (when serotype was found to be S. Typhimurium) and tested for susceptibility to a panel of antibiotics. Salmonella was isolated from 2% of faecal, 2% of rumen and 7.6% of carcass samples. Salmonella was most frequently isolated from samples taken during the period August to October. S. Dublin was isolated from 72% of positive samples. S. Agona and S. Typhimurium definitive type (DT)104 were each isolated from 14% of positive samples. All S. Typhimurium DT104 isolates were resistant to ampicillin, chloramphenicol, streptomycin, sulphafurazole and tetracycline (ACSSuT). On occasion, from a single animal, the same serotype was isolated from more than one sample (i.e. faeces and rumen; faeces and carcass; rumen and carcass; faeces, rumen and carcass). CONCLUSIONS: Salmonella is present in cattle at slaughter and on beef carcasses at an Irish abattoir, with a higher frequency of occurrence during the period August to October. Most isolates from the study are not commonly associated with human clinical infection, with the exception of S. Typhimurium DT104 (R-type ACSSuT). SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides epidemiological data that is necessary for the understanding of beef as a source of human Salmonella infection.     

Utilization of carbonate and ammonia-based treatments to eliminate Escherichia coli O157:H7 and Salmonella Typhimurium DT104 from cattle manure

AIMS: The objective of this study was to investigate alkaline treatments of cattle manure to kill coliforms, Escherichia coli O157:H7 and Salmonella Typhimurium DT104 based on their inhibition by carbonate ion and ammonia. METHODS AND RESULTS: Pure cultures of S. Typhimurium DT104 and E. coli O157:H7 strains were treated with sodium carbonate and ammonia to determine threshold inhibitory concentrations. Fresh cattle manure samples were inoculated with the same strains and their survival was determined after addition of sodium hydroxide, ammonium sulphate, sodium carbonate and/or urea. Control of CO and NH3 concentrations in manure by pH adjustment to 9.5 with sodium hydroxide to more than 5 and 30 mmol l-1, respectively, killed more than 106 cells g-1 in 7 days. Addition of sodium carbonate enhanced the killing effect of NaOH by increasing the CO and NH3 concentrations. Addition of 100 mmol l-1 urea, produced high levels of CO and NH3 and decreased all bacterial counts by at least 106 cells g-1 after 7 days. CONCLUSIONS: Reduction of food-borne pathogens in manure can be achieved by a combination of high concentrations of CO and NH3 which are pH-dependent parameters. SIGNIFICANCE AND IMPACT OF STUDY: Addition of urea could provide a simple manure treatment by combining both antimicrobial factors.     

Evaluation of the pH-dependent,stationary-phase acid tolerance in Listeria monocytogenes and Salmonella Typhimurium DT104 induced by culturing in media with 1% glucose: a comparative study with Escherichia coli O157:H7

AIMS: To comparatively evaluate the adaptive stationary-phase acid tolerance response (ATR) in food-borne pathogens induced by culturing in glucose-containing media, as affected by strain variability and antibiotic resistance, growth temperature, challenge pH and type of acidulant. METHODS AND RESULTS: Antibiotic resistant or sensitive strains of Listeria monocytogenes, Salmonella including S. Typhimurium DT104, and Escherichia coli O157:H7 were cultured (30 degrees C for 24 h; 10 degrees C for up to 14 days) in trypticase soya broth with yeast extract (TSBYE) with 1% or without glucose to induce or prevent acid adaptation, respectively. Cultures were subsequently exposed to pH 3.5 or 3.7 with lactic or acetic acid at 25 degrees C for 120 min. Acid-adapted cultures were more acid tolerant than nonadapted cultures, particularly those of L. monocytogenes and Salmonella. No consistent, positive or negative, influence of antibiotic resistance on the pH-inducible ATR or acid resistance (AR) was observed. Compared with 30 degrees C cultures, growth and acid adaptation of L. monocytogenes and S. Typhimurium DT104 at 10 degrees C markedly reduced their ATR and AR in stationary phase. E. coli O157:H7 had the greatest AR, relying less on acid adaptation. A 0.2 unit difference in challenge pH (3.5-3.7) caused great variations in survival of acid-adapted and nonadapted cells. CONCLUSIONS: Culturing L. monocytogenes and Salmonella to stationary phase in media with 1% glucose induces a pH-dependent ATR and enhances their survival to organic acids; thus, this method is suitable for producing acid-adapted cultures for use in food challenge studies. SIGNIFICANCE AND IMPACT OF THE STUDY: Bacterial pathogens may become acid-adapted in foods containing glucose or other fermentable carbohydrates. Low storage temperatures may substantially decrease the stationary-phase ATR of L. monocytogenes and S. Typhimurium DT104, but their effect on ATR of E. coli O157:H7 appears to be far less dramatic.     

Effect of quinolone treatment on selection and persistence of quinolone-resistant Escherichia coli in swine faecal flora

AIMS: To study the effect of oral administration of a quinolone on emergence of resistance in an indicator bacterial species from faecal flora. METHODS AND RESULTS: Quinolone resistance was studied in Escherichia coli obtained from the faecal contents of pigs housed in nine commercial farrow-to-finish herds in France after administration of flumequine to sows. The percentage of quinolone-resistant E. coli increased in the faeces of sows after administration of flumequine (mean 21.78% at day 7 vs 6.42% before treatment for nalidixic acid) and then decreased (mean 12.6 and 10.4 at days 30 and 60, respectively for nalidixic acid), being not significantly different from initial values 1 month post-treatment. In young pigs, the proportion of resistant strains was lower and decreased over rearing period. Moreover, changes over time of both total E. coli and the proportion of resistant bacteria exhibited great inter-individual variability. CONCLUSIONS: Restoration of susceptible faecal flora occurred within 2 months after flumequine treatment. SIGNIFICANCE AND IMPACT OF THE STUDY: Effect of flumequine treatment of sows on the quinolone resistance of faecal E. coli of both sows and their progeny is noticeable but transitory.     

Acid resistance variability among isolates of Salmonella enterica serovar Typhimurium DT104

AIMS: Acid resistance could be an indicator of virulence since acid resistant strains are able to better survive the human stomach passage and in macrophages. We studied the acid resistance of several Salmonella Typhimurium DT104 strains isolated from food and humans and identified cellular parameters contributing to the enhanced acid resistance of these isolates. METHODS AND RESULTS: Acid resistance was tested in 37 Salmonella enterica Typhimurium serovar DT104 (S. Typhimurium DT104) strains. Acid adaptation at pH 5 followed by exposure for 2 h at pH 2.5 in the 27 human, nine nonhuman, and in two reference strains, revealed strong variation of acid survival. After 2 h at pH 2.5 six strains of S. Typhimurium DT104 were considered high acid resistant as they displayed a level of survival >10%, 14 strains were considered intermediate acid resistant (level of survival was <10% and >0.01%) and 19 strains were considered low acid resistant (level of survival <0.01%). Six strains were selected for further studies and proteomics revealed a relatively high amount of phase 2 flagellin in an acid-sensitive strain and a relatively high amount of the beta component of the H(+)/ATPase in an acid-resistant strain. Two strains were slightly more heat resistant possibly as the result of increased levels of DnaK or GroEL. CONCLUSIONS: A significant difference could be detected between human and food isolates regarding their acid resistance; all high acid-resistant strains were human isolates. SIGNIFICANCE AND IMPACT OF THE STUDY: S. Typhimurium DT104 is known for two decades and has a great impact on human health causing serious food-borne diseases. Our results suggest the existence of a positive correlation between acid resistance and pathogenicity in S. Typhimurium DT104 as all high acid-resistant strains were isolated from humans.     

Surface decontamination of beef inoculated with Salmonella Typhimurium DT104 or Escherichia coli O157:H7 using dry air in a novel heat treatment apparatus

AIMS: To determine the effectiveness of a novel dry air decontamination apparatus in the deactivation of Salmonella serotype Typhimurium DT104 or Escherichia coli O157:H7 on beef surfaces. METHODS AND RESULTS: A laboratory scale dry air decontamination apparatus, capable of producing repeatable and known heating time-temperature cycles on food surfaces was used in decontamination trials. Beef samples were surface inoculated with 7-8 log10CFU cm(-2) of S. Typhimurium DT104 or E. coli O157:H7 and heated at 60, 75, 90 and 100 degrees C using fast and slow heating rates and subsequently held at these temperatures for up to 600 s. A substantial reduction in pathogen numbers was achieved at higher temperatures (90 and 100 degrees C, 4.18-6.06 log10CFU cm(-2)) using both heating rates, but cell survival at these temperatures was also observed. At the lower temperatures, deactivation was small at 60 degrees C in particular it was less than one log unit after 3 min heating. No significant differences were observed when total reductions in pathogen counts were compared for all the temperature/heat up time combinations tested. During slow heating at 90 degrees C, and both heating rates at 100 degrees C, the pattern of deactivation of S. Typhimurium DT104 or E. coli O157:H7 was triphasic. CONCLUSIONS: This study has shown that heating meat surfaces with dry air can achieve substantial reductions in S. Typhimurium DT104 or E. coli O157:H7. As surface decontamination of beef surfaces with dry air had a negative effect on beef colour and appearance, such a decontamination apparatus would be unsuitable for producing meat for retail sale but it could be used to produce safer meat for use in the catering trade. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides researchers and food processors with data on the dynamic changes in S. Typhimurium DT104 and E. coli O157:H7 counts on intact beef surfaces during heating with dry air under realistic (time-varying) temperature conditions.     

Identification of novel Salmonella enterica serovar Typhimurium DT104-specific prophage and nonprophage chromosomal sequences among serovar Typhimurium isolates by genomic subtractive hybridization

Genomic subtractive hybridization was performed between Salmonella enterica serovar Typhimurium LT2 and DT104 to search for novel Salmonella serovar Typhimurium DT104-specific sequences. The subtraction resulted mainly in the isolation of DNA fragments with sequence similarity to phages. Two fragments identified were associated with possible virulence factors. One fragment was identical to irsA of Salmonella serovar Typhimurium ATCC 14028, which is suggested to be involved in macrophage survival. The other fragment was homologous to HldD, an Escherichia coli O157:H7 lipopolysaccharide assembly-related protein. Five selected DNA fragments-irsA, the HldD homologue, and three fragments with sequence similarity to prophages-were tested for their presence in 17 Salmonella serovar Typhimurium DT104 isolates and 27 non-DT104 isolates by PCR. All five selected DNA fragments were Salmonella serovar Typhimurium DT104 specific among the serovar Typhimurium isolates tested. These DNA fragments can be useful for better detection and typing of Salmonella serovar Typhimurium DT104.     

Heterogeneity in expression of lipopolysaccharide by strains of Salmonella enterica serotype Typhimurium definitive phage type 104 and related phage types

AIMS: To investigate lipopolysaccharide (LPS) expression in Salmonella enterica serotype Typhimurium definitive phage type 104 (Salmonella Typhimurium DT104) and related phage types. METHODS AND RESULTS: Isolates were examined for the expression of LPS by SDS-PAGE and silver staining and subtyped by Pulsed Field Gel Electrophoresis (PFGE). The 100 isolates expressed one of two LPS profiles designated A (72%) and B (28%). LPS profiling was able to discriminate between isolates of identical PFGE type. Among 10 groups of outbreak isolates examined, each group was of a single LPS profile: A, 8/10 and B, 2/10. All 10 outbreaks were identical by PFGE analysis. CONCLUSIONS: Isolates of Salmonella Typhimurium DT104 and related phage types expressed one of two distinct LPS profiles. The two LPS profiles appear similar but shifted and in phase with one another, suggesting that the heterogeneity is due to changes in the LPS core region rather than among the repeating oligosaccharide units of the long-chain LPS. SIGNIFICANCE AND IMPACT OF THE SUTDY: LPS profiling provides a useful adjunct to PFGE and other molecular methods for the subtyping of this group of bacteria in epidemiological investigations.     

Transmission routes of Salmonella Typhimurium DT 104 between 14 cattle and pig herds in Denmark demonstrated by molecular fingerprinting

AIMS: Salmonella Typhimurium DT 104 is generally assumed to be spread by contact between live animals, e.g. by trading. The aim of the present study was to assess the importance of other routes of transmission in the dissemination of this bacterium. METHODS AND RESULTS: An outbreak among 14 cattle and pig herds located in a geographically narrow area in Denmark was investigated. Epidemiological information and disease history of the herds was obtained through interviews. Based on this, the hypothesis for horizontal spread was proposed, and these were confirmed by comparison of the pulsed field gel electrophoresis (PFGE) and the plasmid profiles of isolates obtained by continuous sampling over a period of almost 3 years. CONCLUSIONS: The study indicated that other routes might play an important role, than the trading of live animals, in the spread of S. Typhimurium DT 104 among livestock. SIGNIFICANCE AND IMPACT OF THE STUDY: Salmonella Typhimurium DT 104 infected herd might pose a significant risk to herds located within the same geographic area. In advising on how to avoid the spread of this bacterium, factors like person contacts, sharing of equipment and contaminated slurry should be focussed on in addition to infected animals.     

The effect of copper on the death rate of Salmonella typhimurium DT104:30 in food substrates acidified with organic acids

AIMS: The objective of this study was to investigate the effect of copper ions on the survival of Salmonella typhimurium DT104:30 in acidified liquid food substrates. METHODS AND RESULTS: The decimal reduction time (Dvalue) of Salm. typhimurium DT104:30 was determined in acidified liquid pig feed (LPF) and skimmed milk (SM) containing a range of copper concentrations (0-50 ppm). As copper concentration increased, the death rate of Salm. typhimurium DT104:30 increased. In LPF acidified with 150 mmol l-1 lactic acid the presence of 50 ppm copper resulted in a 10-fold increase in the death rate. CONCLUSIONS: The presence of copper salts in acidic liquid food substrates significantly increases the death rate of Salm. typhimurium DT104:30. SIGNIFICANCE AND IMPACT OF THE STUDY: This finding could influence policy on levels of copper in pig feed.     

Antibiotic resistant Escherichia coli and Salmonella in Russian rooks (Corvus frugilegus) wintering in the Czech Republic

AIMS: To characterize antibiotic resistant Escherichia coli and Salmonella isolates in rooks wintering in the Czech Republic. METHODS AND RESULTS: Three hundred and sixty-three faeces samples from rooks were examined for antibiotic resistant Escherichia coli and Salmonella. Altogether 13.7%E. coli isolates were resistant to antimicrobial agents tested. The dominant type of resistance was to tetracycline. Resistant E. coli isolates were examined for antibiotic resistance genes and class 1 integrons. Five of 29 antibiotic resistant isolates possessed the int1 gene. Nine Salmonella isolates (2.5%) were found in rook faeces. All the isolates belonged to serotype Salmonella enterica serovar Enteritidis phage type PT8 and PT23. CONCLUSIONS: The study suggests that rooks can be infected by antibiotic resistant E. coli and Salmonella isolates, probably reflecting the presence of such isolates in their sources of food and/or water in the environment. SIGNIFICANCE AND IMPACT OF THE STUDY: Rooks can serve as reservoirs and vectors of antibiotic resistant E. coli and Salmonella isolates and potentially transmit these isolates over long distances.     

Efficacy of solar disinfection of Escherichia coli, Shigella flexneri, Salmonella Typhimurium and Vibrio cholerae

AIMS: To determine the efficacy of solar disinfection (SODIS) for enteric pathogens and to test applicability of the reciprocity law. METHODS AND RESULTS: Resistance to sunlight at 37 degrees C based on F99 values was in the following order: Salmonella Typhimurium>Escherichia coli>Shigella flexneri>Vibrio cholerae. While F90 values of Salm. Typhimurium and E. coli were similar, F99 values differed by 60% due to different inactivation curve shapes. Efficacy seemed not to be dependent on fluence rate for E. coli stationary cells. Sensitivity to mild heat was observed above a temperature of 45 degrees C for E. coli, Salm. Typhimurium and Sh. flexneri, while V. cholerae was already susceptible above 40 degrees C. CONCLUSIONS: Salmonella Typhimurium was the most resistant and V. cholerae the least resistant enteric strain. The reciprocity law is applicable for stationary E. coli cells irradiated with sunlight or artificial sunlight. SIGNIFICANCE AND IMPACT OF THE STUDY: Escherichia coli might not be the appropriate indicator bacterium to test the efficacy of SODIS on enteric bacteria and the physiological response to SODIS might be different among enteric bacteria. The applicability of the reciprocity law indicates that fluence rate plays a secondary role in SODIS efficacy. Stating inactivation efficacy with T90 or F90 values without showing original data is inadequate for SODIS studies.     

Immunoassays to detect the serum antibody response of cattle to infection with Salmonella Typhimurium definitive type 104 and following vaccination with Bovivac S

AIMS: To use ELISA and immunoblotting assays to examine the serum antibody response of cattle infected with Salmonella Typhimurium DT104 and following vaccination with Bovivac S. METHODS AND RESULTS: Three hundred and twenty-nine cattle, including 16 shedding multiresistant Salmonella Typhimurium DT104, were screened for serum antibodies binding to O=1, 4, 5, 12 lipopolysaccharide (LPS) antigens before and after vaccination with Bovivac S. Sera with an ELISA reading of 0.9A405 or above were shown to contain antibodies, of the IgG-class only, to the LPS of Salmonella Typhimurium using immunoblotting. Prior to vaccination, only 11 cattle had serum IgG-class antibodies to the O=4, 5 LPS antigens, and of these one also had antibodies to outer membrane proteins and H=i flagellar antigens. Following vaccination, 87 out of 315 cattle developed serum antibodies to the LPS of Salmonella Typhimurium. CONCLUSIONS: Evidence of infection of cattle with Salmonella Typhimurium was readily obtained with an LPS-based ELISA in association with an immunoblotting procedure, supplementing existing bacteriological procedures. This enabled the detection of an increase in the number of cattle with serum antibodies to Salmonella Typhimurium LPS following vaccination with Bovivac S. SIGNIFICANCE AND IMPACT OF THE STUDY: The immunoassays described provided evidence of infection with Salmonella Typhimurium and served as a valuable adjunct to established bacteriology.     

Contamination of Pig Carcasses at Two Abattoirs by Escherichia coli with Special Reference to O-serotypes and Antibiotic Resistance

Evidence of the source of carcass contamination of pigs at slaughter was obtained by determining presumptive coliform counts on faeces and on carcass surfaces, and comparing the O-serotypes and antibiotic sensitivity patterns of Escherichia coli from both sites. All of the 16 pig carcasses from the slaughter line of a commercial abattoir were contaminated with presumptive coliform bacilli on most sites examined; the carcasses of six out of eight pigs slaughtered at the Meat Research Institute (MRI) abattoir were also contaminated, but only small numbers of coliforms could be detected on a few of the sites. The proportion of O-serotypes of E. coli present in faeces which were also detected on carcass surfaces, indicating faecal contamination, varied between 0 and 8.6% in MRI slaughtered pigs but reached 66.6% in one group of commercially slaughtered pigs. O-serotypes found on carcass surfaces but not in the faeces of the pigs, were used as an indication of environmental contamination and this was very evident in the commercially slaughtered pigs. A high proportion of E. coli O-serotypes in the gut were resistant to antibiotics and these were also often found on the carcass surface and, since the range of O-serotypes in the pig is similar to that reported in man, the pig must be considered to be a potential reservoir of antibiotic resistant E. coli for man.     

Antimicrobial resistance of Salmonella spp. from pigs at slaughter in Spain in 1993 and 2001

AIM: To compare the incidence of antimicrobial resistance among Salmonella serotypes isolated in a pig slaughterhouse in Zaragoza (Spain) during 1993 and 2001. METHODS AND RESULTS: A total of 168 isolates representing 10 serotypes were examined by disc diffusion method using 17 antibiotics. Data showed that the majority of the strains were resistant to streptomycin (97%), sulfadiazine (93.4%) and tetracycline (83.3%). A large proportion of the collection was multidrug resistant (MDR, resistance to four or more antibiotics) with a greater incidence in 2001. The findings imply an increasing incidence of MDR amongst S. Typhimurium, and all S. Typhimurium-definitive phage type (DT) 104 isolates were resistant to ampicillin, chloramphenicol, streptomycin, sulphonamide and tetracycline (R-ACSSuT). This resistance phenotype had spread among other phage and serotypes. Salmonella Ohio was also a MDR serotype and this is not a serotype normally associated with drug resistance. CONCLUSIONS: A large proportion of the strains were MDR and this showed that pork products could be a potential vehicle of MDR Salmonella food-borne infections. SIGNIFICANCE AND IMPACT OF THE STUDY: The findings may have significant public health consequences and could contribute to the development of useful practices aimed at limiting the transmission of MDR Salmonella serotypes through the food chain.     

Genomic relationship of Salmonella enterica serovar typhimurium DT104 isolates from Korea and the United States

Salmonella enterica serovar Typhimurium DT104 (Salmonella Typhimurium DT104 or DT104) has been emerging as a common pathogen for human in Korea since 1997. In order to compare the genomic relationship and to search for the dominant strains in Korea, we conducted pulsed-field gel electrophoresis (PFGE) and IS200 fingerprinting of 25 epidemiological unrelated isolates from human and animals from Korea and cattle from America. Two Salmonella Typhimurium DT104 isolates from human in Korea and all 8 isolates from American cattle had indistinguishable patterns from the PFGE and IS200 fingerprinting but multidrug-resistant Salmonella Typhimurium isolates, including DT104, from Korean animals had diverse genetic patterns. The data suggest that a dominant DT104 strain might have circulated between Korean and American cattle and that it had a high level of clonality.     

Multiple-locus variable-number tandem repeat analysis of Salmonella enterica subsp. enterica serovar Typhimurium: comparison of isolates from pigs, poultry and cases of human gastroenteritis

AIMS: Pulsed-field gel electrophoresis (PFGE) and variable number tandem repeat (VNTR) profiles of 195 epidemiologically unrelated Salmonella Typhimurium strains isolated in 1997-2004 from pigs were analysed and the results compared to establish the discriminatory ability of each method. In order to investigate the epidemiology of S. Typhimurium from different populations, the VNTR profiles from pigs were compared with those obtained from 190 S. Typhimurium strains isolated from poultry and 186 strains isolated from human cases of gastroenteritis. METHODS AND RESULTS: A total of 195 strains of S. Typhimurium were tested by PFGE and VNTR. For PFGE, the restriction enzyme XbaI was used, and for VNTR, the number of repeats at five loci (STTR 9, 5, 6, 10pl and 3) were counted and assigned an allele number based on an established VNTR scheme. The results obtained showed improved discrimination of VNTR when compared with PFGE with 34 PFGE profiles identified compared with 96 different VNTR profiles for the pig isolates and 56 different VNTR types within the most common PFGE type. Within the three different populations, VNTR showed distinct subpopulations of VNTR type related not only to source, but also demonstrated common VNTR types within samples obtained from humans, poultry and pigs, especially in strains of phage type DT104. CONCLUSIONS: VNTR has taken the discrimination to a further level than that obtained through PFGE, and demonstrated an overlap in the genetic diversity of isolates tested across the three different populations, confirming previous suggestions that animals have an involvement in the dissemination of S. Typhimurium through the food chain. SIGNIFICANCE AND IMPACT OF THE STUDY: Salmonella Typhimurium remains an important concern as a food-borne zoonotic agent. The VNTR strategy described provides an accurate method of tracing strain dissemination, and adds a further level of discrimination to the PFGE type, providing potential benefits to epidemiological studies and the possibility of deciphering source attribution of cases.