Successful treatment of intractable epilepsy with ketogenic diet therapy in twins with ALG3-CDG.

BACKGROUND: Congenital disorders of glycosylation (CDG) is a heterogeneous group of congenital metabolic diseases with multisystem clinical involvement. ALG3-CDG is a very rare subtype with only 24 cases reported so far. CASE: Here, we report two siblings with dysmorphic features, growth retardation, microcephaly, intractable epilepsy, and hemangioma in the frontal, occipital and lumbosacral regions. RESULTS: We studied two siblings by whole exome sequencing. A pathogenic variant in ALG3 (NM_005787.6: c.165C > T; p.Gly55=) that had been previously associated with congenital glycolysis defect type 1d was identified. Their intractable seizures were controlled by ketogenic diet. CONCLUSION: Although prominent findings of growth retardation and microcephaly seen in our patients have been extensively reported before, presence of hemangioma is a novel finding that may be used as an indication for ALG3-CDG diagnosis. Our patients are the first reported cases whose intractable seizures were controlled with ketogenic diet. This report adds ketogenic diet as an option for treatment of intractable epilepsy in ALG3-CDG.

Microbial dynamics in mixed culture biofilms of bacteria surviving sanitation of conveyor belts in salmon-processing plants.

AIMS: The microbiota surviving sanitation of salmon-processing conveyor belts was identified and its growth dynamics further investigated in a model mimicking processing surfaces in such plants. METHODS AND RESULTS: A diverse microbiota dominated by Gram-negative bacteria was isolated after regular sanitation in three salmon processing plants. A cocktail of 14 bacterial isolates representing all genera isolated from conveyor belts (Listeria, Pseudomonas, Stenotrophomonas, Brochothrix, Serratia, Acinetobacter, Rhodococcus and Chryseobacterium) formed stable biofilms on steel coupons (12°C, salmon broth) of about 10(9) CFU cm(-2) after 2 days. High-throughput sequencing showed that Listeria monocytogenes represented 0·1-0·01% of the biofilm population and that Pseudomonas spp dominated. Interestingly, both Brochothrix sp. and a Pseudomonas sp. dominated in the surrounding suspension. CONCLUSIONS: The microbiota surviving sanitation is dominated by Pseudomonas spp. The background microbiota in biofilms inhibit, but do not eliminate L. monocytogenes. SIGNIFICANCE AND IMPACT OF THE STUDY: The results highlights that sanitation procedures have to been improved in the salmon-processing industry, as high numbers of a diverse microbiota survived practical sanitation. High-throughput sequencing enables strain level studies of population dynamics in biofilm.

The effects of different hygiene procedures in reducing bacterial contamination in a model domestic kitchen.

AIMS: Few studies have compared the effectiveness of hygienic cleaning under simulated use conditions. This study compares commonly used and novel cleaning methods for food contact and hand contact surfaces in kitchens. METHODS AND RESULTS: We report results from two surveys on Norwegian consumers' cleaning procedures. Laboratory models involving cutting boards, tap handles and mobile phones contaminated with Escherichia coli and Staphylococcus aureus were used to compare the hygiene efficacy of commonly used cleaning methods together with new technologies (sprays, single-use wipes, and chlorine-based disinfectants). Commonly used cleaning methods produced a mean log10 reduction (LR) in contamination of 1.5-2.5. The efficacy could be improved by drying or including a disinfection step (mean LR 3.1-4.6). Cleaning of mobile phones was common and was improved by including humidity (1.5-1.9 mean LR). CONCLUSIONS: In many situations, traditional methods used by consumers may be sufficient to hygienically clean surfaces. However, in some situations, such as where there are infected or immune-compromised individuals, or where high risk foods are being handled, hygiene practices resulting in higher LR should be recommended. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates that data from models simulating use conditions are required to estimate the effectiveness of detergent-based removal practices and how these can be enhanced by inactivation processes such as drying and disinfection to ensure that contamination from food-borne pathogens is reduced to acceptable levels to prevent infection transmission.

The persistence of Salmonella following desiccation under feed processing environmental conditions: a subject of relevance.

UNLABELLED: Although Salmonella persistence has been predominantly linked to biofilm formation, the physiological state of Salmonella should also be considered as a possible pathway for persistence and survival in the feed industry. Hence, the purpose of this study was to assess the extent of viability of Salmonella cells through long-term desiccation periods under conditions typically found in feed processing environments, and whether these same cells could resuscitate and cause salmonellosis in vivo. We showed that upon desiccation, Salmonella Agona, a representative feed industry isolate and Salmonella Typhimurium ATCC 14028, a laboratory strain, were induced into a nonculturable state at 35 and 85% relative humidity conditions, at defined temperatures of 30 and 12°C, respectively. Although the reduction in culturable cells was more than 6 log10 , metabolic activity was found in more than 1% of the population. Desiccation-induced nonculturable Salm. Typhimurium could not be revived and were nonvirulent in a mouse model following infection through oral gavage. These results suggest that the specific conditions for reviving nonculturable Salmonella after long periods of desiccation are yet to be fully identified. The need for mapping key factors involved in the persistence of Salmonella would help better detect it and improve feed safety measures. SIGNIFICANCE AND IMPACT OF THE STUDY: While Salmonella has been shown to persist for years in feed processing environments, it is still unknown how temperature and humidity affect the persistence of Salmonella cells over time in terms of their metabolic states and cultivability. Here, we show that long-term exposure to feed processing environmental conditions induces Salmonella into a nonculturable state even though about 1% of the population remains metabolically active. This has significant implications when monitoring Salmonella from the environment which could yield false-negative results using conventional pre-enrichment detection methods.

Predicting the concentration of verotoxin-producing Escherichia coli bacteria during processing and storage of fermented raw-meat sausages.

A model to predict the population density of verotoxigenic Escherichia coli (VTEC) throughout the elaboration and storage of fermented raw-meat sausages (FRMS) was developed. Probabilistic and kinetic measurement data sets collected from publicly available resources were completed with new measurements when required and used to quantify the dependence of VTEC growth and inactivation on the temperature, pH, water activity (aw), and concentration of lactic acid. Predictions were compared with observations in VTEC-contaminated FRMS manufactured in a pilot plant. Slight differences in the reduction of VTEC were predicted according to the fermentation temperature, 24 or 34°C, with greater inactivation at the highest temperature. The greatest reduction was observed during storage at high temperatures. A population decrease greater than 6 decimal logarithmic units was observed after 66 days of storage at 25°C, while a reduction of only ca. 1 logarithmic unit was detected at 12°C. The performance of our model and other modeling approaches was evaluated throughout the processing of dry and semidry FRMS. The greatest inactivation of VTEC was predicted in dry FRMS with long drying periods, while the smallest reduction was predicted in semidry FMRS with short drying periods. The model is implemented in a computing tool, E. coli SafeFerment (EcSF), freely available from http://www.ifr.ac.uk/safety/EcoliSafeFerment. EcSF integrates growth, probability of growth, and thermal and nonthermal inactivation models to predict the VTEC concentration throughout FRMS manufacturing and storage under constant or fluctuating environmental conditions.

Microbial background flora in small-scale cheese production facilities does not inhibit growth and surface attachment of Listeria monocytogenes.

The background microbiota of 5 Norwegian small-scale cheese production sites was examined and the effect of the isolated strains on the growth and survival of Listeria monocytogenes was investigated. Samples were taken from the air, food contact surfaces (storage surfaces, cheese molds, and brine) and noncontact surfaces (floor, drains, and doors) and all isolates were identified by sequencing and morphology (mold). A total of 1,314 isolates were identified and found to belong to 55 bacterial genera, 1 species of yeast, and 6 species of mold. Lactococcus spp. (all of which were Lactococcus lactis), Staphylococcus spp., Microbacterium spp., and Psychrobacter sp. were isolated from all 5 sites and Rhodococcus spp. and Chryseobacterium spp. from 4 sites. Thirty-two genera were only found in 1 out of 5 facilities each. Great variations were observed in the microbial background flora both between the 5 producers, and also within the various production sites. The greatest diversity of bacteria was found in drains and on rubber seals of doors. The flora on cheese storage shelves and in salt brines was less varied. A total of 62 bacterial isolates and 1 yeast isolate were tested for antilisterial activity in an overlay assay and a spot-on-lawn assay, but none showed significant inhibitory effects. Listeria monocytogenes was also co-cultured on ceramic tiles with bacteria dominating in the cheese production plants: Lactococcus lactis, Pseudomonas putida, Staphylococcus equorum, Rhodococcus spp., or Psychrobacter spp. None of the tested isolates altered the survival of L. monocytogenes on ceramic tiles. The conclusion of the study was that no common background flora exists in cheese production environments. None of the tested isolates inhibited the growth of L. monocytogenes. Hence, this study does not support the hypothesis that the natural background flora in cheese production environments inhibits the growth or survival of L. monocytogenes.

Effects of post-processing treatments on sensory quality and Shiga toxigenic Escherichia coli reductions in dry-fermented sausages.

The effects of post-processing treatments on sensory quality and reduction of Shiga toxigenic Escherichia coli (STEC) in three formulations of two types of dry-fermented sausage (DFS; salami and morr) were evaluated. Tested interventions provided only marginal changes in sensory preference and characteristics. Total STEC reductions in heat treated DFS (32°C, 6days or 43°C, 24h) were from 3.5 to >5.5 log from production start. Storing of sausages (20°C, 1month) gave >1 log additional STEC reduction. Freezing and thawing of sausages in combination with storage (4°C, 1month) gave an additional 0.7 to 3.0 log reduction in STEC. Overall >5.5 log STEC reductions were obtained after storage and freezing/thawing of DFS with increased levels of glucose and salt. This study suggests that combined formulation optimisation and post-process strategies should be applicable for implementation in DFS production to obtain DFS with enhanced microbial safety and high sensory acceptance and quality.

Heterogeneity in resistance to food-related stresses and biofilm formation ability among verocytotoxigenic Escherichia coli strains.

This study assessed the resistance of ten verocytotoxigenic Escherichia coli (VTEC) isolates of commonly encountered serogroups/-types and two non-pathogenic E. coli strains to various food-related stresses (acid, alkaline, heat and high hydrostatic pressure treatments) and their biofilm formation ability. In addition, the global changes in the cellular composition in response to the exposure to these adverse environments were monitored by Fourier Transform Infrared (FT-IR) spectroscopy for two of the strains. Large inter-strain variations in stress resistance were observed. The most tolerant strains belonged to serogroup O157 which included both the O157:H7 type strain EDL933 and a representative isolate of the sorbitol fermenting O157:H- VTEC clone (strain MF3582). Strain C-600, a non-pathogenic laboratory strain, was sensitive to multiple stresses. Although wide variation in biofilm-forming ability was observed among VTEC isolates, no consistent relationships between biofilm-forming ability and capacity to withstand stress exposures were found. Analysis of the allelic status of the rpoS gene, involved in the general stress response of stationary-phase cells, allowed detection of loss-of-function mutations for two strains, E218/02 and MF2411, both of them showing as common features a high sensitivity to alkaline and heat treatments and a poor ability to form mature biofilms. Evidences found in this study confirm rpoS as a highly mutable gene in nature, and suggest its relevance not only for the mount of an active stress response but also for the establishment of mature biofilm communities. Our findings contribute to increase the knowledge on the resistance of VTEC to environmental stresses commonly encountered in the food chain, which can lead to improved strategies for preventing VTEC infections.

Reduction of verotoxigenic Escherichia coli by process and recipe optimisation in dry-fermented sausages.

Outbreaks of verotoxigenic Escherichia coli (VTEC) linked to dry-fermented sausages (DFSs) have emphasized the need for DFS manufacturers to introduce measures to obtain enhanced safety and still maintain the sensory qualities of their products. To our knowledge no data have yet been reported on non-O157:H7 VTEC survival in DFS. Here, the importance of recipe and process variables on VTEC (O157:H7 and O103:H25) reductions in two types of DFS, morr and salami, was determined through three statistically designed experiments. Linear regression and ANOVA analyses showed that no single variable had a dominant effect on VTEC reductions. High levels of NaCl, NaNO(2), glucose (low pH) and fermentation temperature gave enhanced VTEC reduction, while high fat and large casing diameter (a(w)) gave the opposite effect. Interaction effects were small. The process and recipe variables showed similar effects in morr and salami. In general, recipes combining high batter levels of salt (NaCl and NaNO(2)) and glucose along with high fermentation temperature that gave DFS with low final pH and a(w), provided approximately 3 log(10) reductions compared to approximately 1.5 log(10) reductions obtained for standard recipe DFS. Storage at 4°C for 2months provided log(10) 0.33-0.95 additional VTEC reductions and were only marginally affected by recipe type. Sensory tests revealed only small differences between the various recipes of morr and salami. By optimisation of recipe and process parameters, it is possible to obtain increased microbial safety of DFS while maintaining the sensory qualities of the sausages.

Application of high pressure processing to reduce verotoxigenic E. coli in two types of dry-fermented sausage.

The effect of high pressure processing (HPP) on the survival of verotoxigenic Escherichia coli (VTEC) in two types of Norwegian type dry-fermented sausages was studied. Two different types of recipes for each sausage type were produced. The sausage batter was inoculated with 6.8 log(10) CFU/g of VTEC O103:H25. After fermentation, drying and maturation, slices of finished sausages were vacuum packed and subjected to two treatment regimes of HPP. One group was treated at 600 MPa for 10 min and another at three cycles of 600 MPa for 200 s per cycle. A generalized linear model split by recipe type showed that these two HPP treatments on standard recipe sausages reduced E. coli by 2.9 log(10) CFU/g and 3.3 log(10) CFU/g, respectively. In the recipe with higher levels of dextrose, sodium chloride and sodium nitrite E. coli reduction was 2.7 log(10) CFU/g in both treatments. The data show that HPP has a potential to make the sausages safer and also that the effect depends somewhat on recipe.

Reduction of verotoxigenic Escherichia coli by process and recipe optimisation in dry-fermented sausages.

Outbreaks of verotoxigenic Escherichia coli (VTEC) linked to dry-fermented sausages (DFSs) have emphasized the need for DFS manufacturers to introduce measures to obtain enhanced safety and still maintain the sensory qualities of their products. To our knowledge no data have yet been reported on non-O157:H7 VTEC survival in DFS. Here, the importance of recipe and process variables on VTEC (O157:H7 and O103:H25) reductions in two types of DFS, morr and salami, was determined through three statistically designed experiments. Linear regression and ANOVA analyses showed that no single variable had a dominant effect on VTEC reductions. High levels of NaCl, NaNO(2), glucose (low pH) and fermentation temperature gave enhanced VTEC reduction, while high fat and large casing diameter (a(w)) gave the opposite effect. Interaction effects were small. The process and recipe variables showed similar effects in morr and salami. In general, recipes combining high batter levels of salt (NaCl and NaNO(2)) and glucose along with high fermentation temperature that gave DFS with low final pH and a(w), provided approximately 3 log(10) reductions compared to approximately 1.5 log(10) reductions obtained for standard recipe DFS. Storage at 4 degrees C for 2 months provided log(10) 0.33-0.95 additional VTEC reductions and were only marginally affected by recipe type. Sensory tests revealed only small differences between the various recipes of morr and salami. By optimisation of recipe and process parameters, it is possible to obtain increased microbial safety of DFS while maintaining the sensory qualities of the sausages.

Factors affecting survival of Shigatoxin-producing Escherichia coli on abiotic surfaces.

Shigatoxin-producing Escherichia coli (STEC) causes severe infections, and has been the cause of a number of foodborne outbreaks. Knowledge on the survival of STEC is crucial in order to limit the risk of cross contamination and transfer of STEC to food during processing. In this study survival of STEC and non-STEC on surfaces under various humidities, temperatures and in the presence of different types of soil was investigated. A model system with controlled relative humidity and temperature was established by using saturated salt solutions. All the 12 STEC strains had a reduction in viable count during incubation at 70% RH at 12 degrees C. The reduction was 2-3.5 log and 4.5-5.5 log after 1 and 7 days of incubation, respectively. Surviving cells were observed after 19 days of incubation. The STEC strains were more resistant to desiccation than non-STEC strains. STEC survived better at 12 degrees C, compared to 20 degrees C. The survival of STEC was much lower than the survival of a Staphylococcus simulans strain tested, which showed less than 1 log reduction until day 7 at 70% RH at 12 degrees C, while several STEC strains had comparable survival to a Salmonella Agona strain. The survival of two STEC strains tested was highest at 98% RH. The lowest survival was observed at 85% RH, with better survival at drier conditions. Presence of proteins and glucose protected the cells at dry conditions. Two commercial disinfectants tested at in-use concentration had limited effect (0.8-2.5 log reduction) against STEC on stainless steel, especially for cells incubated at high relative humidity (98% RH). STEC surviving on surfaces in the food industry may impose a risk for cross contamination. Cleaning and use of suitable disinfectants will reduce the survival of STEC, but surfaces should be allowed to dry completely since humid conditions will promote the survival and growth of STEC.

Characterization of the bacterial spoilage flora in marinated pork products.

AIMS: To investigate the microbiota in marinated, vacuum-packed pork and to characterize isolated bacteria with regard to their spoilage potential. METHODS AND RESULTS: Laboratory marinated pork meat and commercial products from three Norwegian producers were examined. Lactic acid bacteria dominated in all products at the expiration date. The flora in marinated products was similar only for products from the same plant. Strains of Lactobacillus algidus, Lactobacillus sakei, Lactobacillus curvatus, Carnobacterium divergens, Carnobacterium maltaromaticum, Leuconostoc mesenteroides, Leuconostoc carnosum and Leuconostoc sp. were isolated and tested for their spoilage potential. Samples inoculated with Lact. algidus or Leuc. mesenteroides were rated as most unpleasant by randomly selected people. A sensory panel scored samples with Lact. algidus highest for sour and intense odour. Lactobacillus algidus was found in products from two out of three production plants. Culture-independent DNA isolation confirmed that cultivation on Blood agar at 20 degrees C yielded a representative picture of the total flora in marinated flintsteak. CONCLUSIONS: Lactobacillus algidus may be an important, but underestimated, spoilage organism that needs to be focused on more when spoilage of vacuum-packed meat is considered. SIGNIFICANCE AND IMPACT OF THE STUDY: Routine microbial testing may have to be revised in order to detect spoilage LAB that are unable to grow under currently used conditions.

Molecular epidemiology of Salmonella spp. isolates from gulls, fish-meal factories, feed factories, animals and humans in Norway based on pulsed-field gel electrophoresis.

The molecular epidemiology of 98 isolates of Salmonella serovar Agona (n = 27), S. Montevideo (n = 42) and S. Senftenberg (n = 29) from wild-living gulls, fish-meal factories, feed factories, humans and domestic animals was investigated using pulsed-field gel electrophoresis (PFGE) and computerized numerical analysis. Two of the S. Agona profiles were identified both in gulls and in two of the factories. In addition, one of these profiles was detected in two infected poultry farms. Two of the S. Montevideo profiles were also identified both in gulls and in two of the factories, and one of these profiles was observed in a human isolate. Four factories shared an identical S. Senftenberg profile. The S. Senftenberg profile found in gulls was not identified in any other source investigated. The presence of isolates with identical PFGE profiles indicates potential epidemiological links between different factories, as well as between gulls and factories.

Outbreak of Salmonella Livingstone infection in Norway and Sweden due to contaminated processed fish products.

In Europe, the number of reported sporadic human cases of Salmonella Livingstone infection is low, and outbreaks are rare. We report the largest S. Livingstone outbreak described in the literature having an identified source of infection. In February 2001, an increased incidence of infection caused by S. Livingstone was observed in Norway and Sweden. By July 2001, 44 cases were notified in Norway and 16 in Sweden. The median age was 63 years, and 40 were women. There were three deaths, and 22 patients were hospitalized. Based on standardized questionnaires and retrospective studies of S. Livingstone strains in Norway and Sweden, food items with egg powder were suspected, and S. Livingstone was subsequently recovered from a processed fish product at the retail level. Analysis by pulsed-field gel electrophoresis documented that isolates from the fish product belonged to the same clone as the outbreak strain.

Molecular epidemiology and population genetics of Salmonella subspecies diarizonae in sheep in Norway and Sweden.

Fifty-four isolates of Salmonella enterica subsp. diarizonae (IIIb) in Norway, Sweden, England, the United States, France and Australia were characterized by pulsed-field gel electrophoresis (PFGE). This study focuses on serovar 61:k:1,5,(7) [S. IIIb 61:k:1,5,(7)] isolated from sheep. Digestion of the bacterial DNA with restriction enzyme XhaI yielded 15 distinct PFGE profiles comprising 12-16 fragments in the range 48.5-630.5 kbp. Four different profiles were identified in Norwegian sheep isolates and a single profile in Swedish isolates. The spatial and temporal distribution of profiles is discussed.

Isolation of Shiga toxin-producing Escherichia coli O103 from sheep using automated immunomagnetic separation (AIMS) and AIMS-ELISA: sheep as the source of a clinical E. coli O103 case?

AIMS: To investigate whether a sheep flock was the original reservoir of a Shiga toxin-producing Escherichia coli (STEC) O103 strain causing a clinical human case and to compare the two diagnostic methods automated immunomagnetic separation (AIMS) and AIMS-ELISA. METHODS AND RESULTS: AIMS detected Escherichia coli O103 in 36.5% of the samples and AIMS-ELISA detected E. coli O103 in 52.1% of the samples. Polymerase chain reaction detected stx1 and eae in three of 109 E. coli O103 isolates. Pulsed field gel electrophoresis showed that the sheep and human STEC O103 were characterized by distinctly different profiles. CONCLUSIONS: The sheep flock was shown to carry STEC O103, although an association between the sheep flock and the clinical human case could neither be proven nor eliminated. Substantial agreement was found between AIMS and AIMS-ELISA, but AIMS-ELISA was less time consuming and resulted in a higher recovery of E. coli O103. SIGNIFICANCE AND IMPACT OF THE STUDY: The study shows that sheep may be carriers of STEC that are associated with human disease and that the methods described can be used to increase the sensitivity of STEC detection.

Molecular epidemiology of Salmonella typhimurium isolates from human sporadic and outbreak cases.

The molecular epidemiology of a representative collection of sporadic foreign and domestically acquired Salmonella Typhimurium (S. Typhimurium) isolates from Norwegian patients in 1996-9 was studied by numerical analysis of pulsed-field gel electrophoresis (PFGE) profiles. Three subclusters (E5, F1 and G1) comprised 47% of the 102 sporadic isolates investigated and 45% of the domestically acquired isolates fell in subclusters E5 and F1. Distinct seasonal and geographic variations were evident for these strains which have been responsible for both local outbreaks (E5) and a national epidemic (F1) where salmonella-infected hedgehogs and birds constituted the suggested primary source of infection. Subcluster G1 was dominated by imported multi-resistant definitive type (DT) 104 isolates. All multi-resistant isolates contained integron-associated gene cassette-structures. This study presents valuable information on the relative significance, geographic distribution and antibiotic resistance features of distinct S. Typhimurium clones causing human salmonellosis among Norwegians.

Escherichia coli O157:H7 in faeces from cattle, sheep and pigs in the southwest part of Norway during 1998 and 1999.

During a 2-year period from January 1998 to December 1999, intestinal content from 1541 cattle, 665 sheep and 1976 pigs were analysed for Escherichia coli O157:H7 using the immunomagnetic separation procedure. The animals originated from 848, 605 and 832 herds from the southwest part of Norway, respectively. E. coli O157:H7 was present in three samples from cattle from different herds, giving a herd prevalence of 0.35% and an animal prevalence of 0.19%. From pigs, E. coli O157:H7 was isolated from two pigs from different herds, giving a herd prevalence of 0.24% and an animal prevalence of 0.1%. A follow-up study revealed another positive testing pig from one of these herds. E. coli O157:H7 was not found from any of the 665 investigated sheep. By PCR analysis, all six E. coli O157:H7 isolates were shown to contain the genes encoding Shiga toxin 2 (stx2), the intimin protein (eae) and the H7 flagellum (fliC-H7). One of the cattle isolates also harboured the Shiga toxin 1 encoding (stx1) gene. The six isolates were differentiated into three pulse-field gel electrophoresis profiles. The results indicate that the occurrence of E. coli O157:H7 in cattle, sheep and pigs in the southwest part of Norway is low compared to other European countries.

Genetic linkage between resistance to quaternary ammonium compounds and beta-lactam antibiotics in food-related Staphylococcus spp.

Little is known about the occurrence of antimicrobial resistance determinants in staphylococci isolated from food and food processing industries. Quaternary ammonium compound (QAC)-resistant coagulase-negative staphylococci (CNS) isolated from food and food-processing industries were investigated for the presence of genetic determinants (qacA/B and qacC/smr) encoding resistance to the QAC benzalkonium chloride (BC), several antibiotic resistance genes, and staphylococcal insertion sequences IS257 and IS256. Six qacA/B-harboring strains were resistant to penicillin and hybridized to a blaZ probe. The qacA/B and blaZ probes hybridized to plasmids of similar size in three isolates. Molecular and genetic characterization of the 23-kb plasmid (pST6) of Staphylococcus epidermidis St.6 revealed the presence of qacB adjacent to an incomplete beta-lactamase transposon Tn552 encoding the gene cluster blaZ, blaR, and blaI. Sequence analysis of flanking regions and the intergenic region between blaZ and qacB revealed the presence of IS257 downstream of blaZ as well as sin and binR between blaZ and qacB. In the three other BC and penicillin-resistant strains, the qacA/B and blaZ genes were located on separate plasmids. A qacC harboring S. epidermidis strain (St.17) also hybridized to tetK (tetracycline resistance) and ermB (erythromycin resistance) genes. The individual genes were located on separate plasmids, suggesting no linkage between QAC and antibiotic resistance determinants. Plasmid-free Staphylococcus aureus RN4220 allowed uptake of the pST6 plasmid DNA, indicating that the resistance genes could potentially be transferred to pathogens under selective stress. In conclusion, presence of both resistance determinants could lead to co-selection during antimicrobial therapy or disinfection in hospitals or in food industries.