Fitness cost associated with resistance to fluoroquinolones is diverse across clones of Klebsiella pneumoniae and may select for CTX-M-15 type extended-spectrum ß-lactamase.

Lowered fitness cost associated with resistance to fluoroquinolones was recently demonstrated to influence the clonal dynamics of methicillin-resistant Staphylococcus aureus (MRSA) in the health care setting. We investigated whether or not a similar mechanism impacts Klebsiella pneumoniae. The fitness of K. pneumoniae isolates from major international hospital clones (ST11, ST15, ST147) already showing high-level resistance to fluoroquinolones and of strains from three minor clones (ST25, ST274, ST1028) in which fluoroquinolone resistance was induced in vitro was tested in a propagation assay. Strains from major clones showed significantly less fitness cost than three of four fluoroquinolone-resistant derivatives of minor clone isolates. In addition, plasmids with CTX-M-15 type extended-spectrum ß-lactamase (ESBL) genes were all retained in both major and minor clone isolates, irrespective of the strains' level of fluoroquinolone resistance, while each plasmid harboring SHV-type ESBLs had been lost during the induction of resistance. Major clone K. pneumoniae strains harbored more amino acid substitutions in the quinolone resistance determining regions (QRDRs) of the gyrA and parC genes than minor clone isolates. The presence of an active efflux system could be demonstrated in all fluoroquinolone-resistant derivatives of originally SHV-producing minor clone isolates but not in any CTX-M-15-producing strain. Further investigations are needed to expand and confirm our findings on a larger sample. In addition, a long-term observation of our ciprofloxacin-resistant minor clone isolates is required in order to elucidate whether or not they are capable of restoring their fitness while concomitantly retaining high minimum inhibitory concentration (MIC) values.

Multidisciplinary investigation of a multicountry outbreak of Salmonella Stanley infections associated with turkey meat in the European Union, August 2011 to January 2013.

Between August 2011 and January 2013, an outbreak of Salmonella enterica serovar Stanley (S. Stanley) infections affected 10 European Union (EU) countries, with a total of 710 cases recorded. Following an urgent inquiry in the Epidemic Intelligence Information System for food- and waterborne diseases (EPIS-FWD) on 29 June 2012, an international investigation was initiated including EU and national agencies for public health, veterinary health and food safety. Two of three local outbreak investigations undertaken by affected countries in 2012 identified turkey meat as a vehicle of infection. Furthermore, routine EU monitoring of animal sources showed that over 95% (n=298) of the 311 S. Stanley isolates reported from animal sampling in 2011 originated from the turkey food production chain. In 2004­10, none had this origin. Pulsed-field gel electrophoresis (PFGE) profile analysis of outbreak isolates and historical S. Stanley human isolates revealed that the outbreak isolates had a novel PFGE profile that emerged in Europe in 2011. An indistinguishable PFGE profile was identified in 346 of 464 human, food, feed, environmental and animal isolates from 16 EU countries: 102 of 112 non-human isolates tested were from the turkey production chain. On the basis of epidemiological and microbiological evidence, turkey meat was considered the primary source of human infection, following contamination early in the animal production chain.

Investigation into an unusual increase of human cases of Salmonella Goldcoast infection in Hungary in 2009.

We describe the outbreak investigation associated with an unusual increase in Salmonella Goldcoast cases in Hungary observed in autumn 2009, which included descriptive and analytical epidemiological studies and microbiological and veterinary investigations. Sixty cases were identified between 1 January 2009 and 1 March 2010, 50 of them from late July 2009 to January 2010. Of 50 S. Goldcoast isolates, 44 showed an indistinguishable pulsed-field gel electrophoresis profile. We conducted a matched case-control study that indicated a statistically significant association between S. Goldcoast infection and the consumption of pork cheese. The majority of cases (seven of nine) reporting consumption of this product belonged to a single family cluster. After removing six cases of this cluster, pork cheese still showed an elevated but non-significant risk for being a case in the univariable analysis (Mantel-Haenszel odds ratio (MH OR): 3.87, 95% confidence interval (CI): 0.38-39.47). A single S. Goldcoast isolate was identified during routine veterinary surveillance activities in 2009 in minced beef from a butcher's shop, originating from an abattoir where also pigs were slaughtered. We conclude that the outbreak was probably due to multiple sources of contaminated meat, probably pork, released on the market over a period of several months in 2009.

Varying fitness cost associated with resistance to fluoroquinolones governs clonal dynamic of methicillin-resistant Staphylococcus aureus.

The purpose of this study was to investigate the impact of fluoroquinolone resistance on the existence and dynamic of MRSA clones. Resistance to ciprofloxacin was induced in strains of community-acquired (CA) MRSA from various sequence types and the fitness cost suffered by mutant derivatives measured in a propagation assay. In addition, the fitness of fluoroquinolone resistant health care-associated (HA) MRSA isolates from major clones prevalent in Hungary were compared with each other and with those of the CA-MRSA derivatives. The genetic background of fluoroquinolone resistance and fitness cost in CA-MRSA was investigated. The fitness cost observed in the CA-MRSA derivatives proved diverse; the derivatives of the ST30-MRSA-IV strain suffered significantly greater fitness cost than those of the ST8-MRSA-IV and ST80-MRSA-IV isolates. Strains from the New York-Japan (ST5-MRSA-II), South German (ST228-MRSA-I) and EMRSA-15 (ST22-MRSA-IV) HA-MRSA clones proved more viable than CA-MRSA derivatives with similar MIC values to ciprofloxacin and HA-MRSA strains from the Hungarian/Brazilian clone (ST239-MRSA-III). Our strains from the New York-Japan, South-German and EMRSA-15 clones seem to have a competitive edge over the tested CA-MRSA isolates in the health care setting. The greater fitness observed in our New York-Japan and South-German strains could account for the replacement by them of the Hungarian/Brazilian clone in Hungary about ten years ago. Alterations in relevant genes were detected. The Ser80 → Phe mutation in the grlA gene may have seriously compromised viability. Surprisingly silent nucleotide substitutions in the grlB gene seemed to impact fitness in derivatives of the ST30-MRSA-IV isolate.

Emergence of a colistin-resistant KPC-2-producing Klebsiella pneumoniae ST258 clone in Hungary.

Nine Klebsiella pneumoniae isolates showing non-susceptibility to carbapenems were collected from three centres in the north-eastern region of Hungary. The minimum inhibitory concentrations (MICs) of antibiotics were determined by Etest. The putative production of a carbapenemase was tested by the modified Hodge test. The presence of bla (KPC) genes was verified by polymerase chain reaction (PCR) and sequencing. Furthermore, molecular typing was performed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). All isolates showed extensively drug-resistant (XDR) phenotype, and of these, eight isolates were highly resistant to colistin. The isolates carried bla (KPC-2), bla (SHV-12), bla (TEM-1) and bla (SHV-11). PFGE analysis of the nine KPC-2-producing Hungarian ST258 K. pneumoniae isolates, two KPC-2-producing Norwegian ST258 isolates and 33 CTX-M-15-producing ST11 isolates revealed the existence of one genetic cluster at an 88% similarity level. The overall results of the PFGE clustering, MLST and the presence of SHV-11 in both ST11 and ST258 suggest that this is the first hyperepidemic clonal complex of multidrug-resistant K. pneumoniae, probably CC258/CC340, possibly undergoing worldwide spread.

Prevalence and characterization of Salmonella infantis isolates originating from different points of the broiler chicken-human food chain in Hungary.

During the 10-month study period Salmonella contamination of broiler houses and the flocks reared in three farms (A, B and C), the slaughter houses where the flocks were slaughtered, as well as the carcass and retail raw meat products originating from them was investigated. In the broiler farm A five consecutive flocks, in the B and C farms one flock was sampled. Environmental samples were taken prior to the introductions. Environmental, drinking water, feed and faecal samples were collected regularly using standard methods. Before and during processing of the flocks, environmental and carcass samples were taken at the abattoirs. Salmonella contamination of the carcass, retail meat, as well as stool samples of farm and abattoir workers and from human illnesses registered in the same period and region were also examined. Isolation, sero-, phage- and antibiotic resistance typing, class 1 integron and plasmid profiling of the strains were performed; their genetic relationship was assessed by PFGE. Although the broiler house and the faecal samples of the 5 flocks of the farm A were negative for Salmonella, S. infantis was isolated from 20-100% of the abattoir carcass samples. The retail raw meat samples were 0-100% S. infantis positive. The environmental samples of farm B were Salmonella negative, but the examined flock was contaminated: S. infantis was identified from 43% of the faecal samples. This serotype was identified in 100% of the carcass and retail raw meat samples. From environmental samples taken before the arrival of the 1-day-old chicks in the broiler house C, S. infantis was cultured. S. infantis prevalence in the faecal samples was 35% and all the carcass and retail raw meat samples were S. infantis contaminated. Altogether 164 S. infantis strains were isolated out of which 145 were further characterized. The vast majority (142/145) of the strains belonged to phage types 217 and 213. All but one were characterized by the nalidixic acid-streptomycin-sulphonamide-tetracycline resistances, had an 885 bp class 1 integron and a large plasmid of > 168 kb in size. The strains showed > or = 88.7% genetic similarity. The results obtained shows that the same multi-drug resistant S. infantis clone was spread from the examined broiler farms contaminating the slaughter and the retail meat and appeared in the human illnesses of the examined region that was earlier detected as the dominant clone characteristic of the broiler and human population of the whole country.

Replacement of methicillin-resistant Staphylococcus aureus clones in Hungary over time: a 10-year surveillance study.

The prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in Hungary has been increasing and is now close to 20% among invasive isolates of S. aureus. In order to understand the evolution of MRSA in Hungary, two collections of isolates were studied: 22 representatives of a collection of 238 MRSA isolates recovered between 1994 and 1998, and a collection of 299 MRSA isolates recovered between 2001 and 2004. The isolates were first characterised by pulsed-field gel electrophoresis (PFGE) and were distributed into 19 different PFGE patterns. Representatives of each pattern were further characterised by spa typing, multilocus sequence typing (MLST) and staphylococcal cassette chromosome mec (SCCmec) typing. The Hungarian clone that was predominant in 1994-1998 (PFGE E, ST239-III) had almost disappeared in 2003-2004, being replaced by the Southern German clone (PFGE B, ST228-I) and the New York/Japan epidemic clone (PFGE A, ST5-II), which represented c. 85% of the 2001-2004 isolates. Thus, this study describes, for the first time, the co-dominance and extensive spread of the New York/Japan clone in a European country.

Virulence and antimicrobial resistance determinants of verotoxigenic Escherichia coli (VTEC) and of multidrug-resistant E. coli from foods of animal origin illegally imported to the EU by flight passengers.

The aim of this study was to reveal phenotype/genotype characteristics of verotoxigenic Escherichia coli (VTEC) and multidrug resistant E. coli in food products of animal origin confiscated as illegal import at Austrian, German and Slovenian airports. VTEC isolates were obtained by using ISO guidelines 16654:2001 for O157 VTEC or ISO/ TS13136:2012 for non-O157 VTEC, with additional use of the RIDASCREEN® Verotoxin immunoassay. The testing of 1526 samples resulted in 15 VTEC isolates (1.0%) primarily isolated from hard cheese from Turkey and Balkan countries. Genotyping for virulence by using a miniaturized microarray identified a wide range of virulence determinants. One VTEC isolate (O26:H46) possessing intimin (eae) and all other essential genes of Locus of Enterocyte Effacement (LEE) was designated as enterohemorrhagic E. coli (EHEC). None of the other VTEC strains belonged to serogroups O157, O145, O111, O104 or O103. VTEC strains harbored either stx(1) (variants stx1(a) or stx(1c)) or st(x2) (variants stx(2a), stx(2b), stx(2a/d) or stx(2c/d)) genes. Pulsed field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) demonstrated high genetic diversity and identified three new sequence types (STs): 4505, 4506 and 4507. Food samples collected from the Vienna airport were also tested for E. coli quantities using the ISO 16649:2001, and for detection of multidrug resistant phenotypes and genotypes. The resulting 113 commensal E. coli isolates were first tested in a pre-screening against 6 selected antimicrobials to demonstrate multidrug resistance. The resulting 14 multidrug resistant (MDR) E. coli isolates, representing 0.9% of the samples, were subjected to further resistance phenotyping and to microarray analyses targeting genetic markers of antimicrobial resistance and virulence. Genotyping revealed various combinations of resistance determinants as well as the presence of class 1, class 2 integrons. The isolates harbored 6 to 11 antibiotic resistance genes as well as 1 to 14 virulence genes. In this panel of 14 MDR E. coli two strains proved to carry CTX-M type ESBLs, and one single isolate was identified as enteropathogenic E. coli (EPEC). In general, isolates carrying a high number of resistance determinants had lower number of virulence genes and vice versa. In conclusion, this first pilot study on the prevalence of VTEC and of MDR/ESBL E. coli in illegally imported food products of animal origin suggests that these strains could represent reservoirs for dissemination of potentially new types of pathogenic and MDR E. coli in Europe.

Effect of precultivation conditions on colicin susceptibility in Escherichia coli.

The effect of 20 colicins and cloacin was studied after various precultivations. Nutrient agar supplemented with subinhibitory concentration of EDTA used for precultivation or elevating the growth-temperature of the inoculum from 37 degrees C to 42 degrees C increased the susceptibility of wild-type (smooth) Escherichia coli strains to the inhibitory action of some colicins. There were great differences among the colicins in respect to these effects. In case of rough mutants, their sensitivities did not change or eventually decrease after EDTA or heat pretreatment. The LPS pattern in SDS-PAGE of smooth cells grown in EDTA-containing nutrient medium changed in some degree towards the rough character. In case of precultivation at 42 degrees C this change was less considerable. It is supposed that both factors applied during precultivation have influence on colicin sensitivity by means of the change of receptor activity caused by LPS modification.

Epidemiology and characterization of animal Salmonella enterica subspecies Enterica serotype typhimurium DT104 in Hungary.

Reports on the internationally emerging significance of multiresistant zoonotic Salmonella in animals and man prompted studies to estimate the significance of multiresistant Salmonella enterica subspecies enterica serotype Typhimurium (S. Typhimurium) phage type DT104 of animal origin in Hungary. A collection of 231 strains (primarily of goose, turkey, poultry and porcine origin from the years 1997-1998) was tested for resistance against 7 selected antibiotics (ampicillin, chloramphenicol, enrofloxacin, nalidixic acid, streptomycin, tetracycline and sulphamethoxazole). Strains with resistance against 3 or more were defined as multiresistant. All strains were phage typed using Felix-Callow's S. Typhimurium phage typing system, and 91 of them (suspect DT104) were also typed according to Anderson's definitive typing (DT) system. In this study, 14% of animal strains from 1997-1998 was classified as DT104, for which turkey, pig and duck seemed to be the main carriers, and the multiresistant non-DT104 strains represented a further 6% of this collection. The prevalence of DT104 was highest among strains of turkey origin (50%), followed by strains of pig (29%), chicken (25%), duck (19%), and goose (3%) origin. The other DT104 related phage types (DT12 and U302) were only detected in the case of 4 strains (2 of porcine, and one each of turkey and of goose origin). The DT104 corresponded to the Felix-Callow types 2/3 or 2c/3 in each case, except in the case of 3 turkey strains where they corresponded to type 35/3. Nalidixic acid resistance was detected in all multiresistant turkey strains and in some of other animal origin but none of these strains were resistant to enrofloxacin. A retrospective analysis (based on the above relationship) indicated that S. Typhimurium strains corresponding to DT104 could be present and increase in the Hungarian farm animal population from about 2% to 20% between 1985 and 1990, in a manner similar to the emergence of human DT104, as reported elsewhere (Pászti et al., 2000). The 91 suspect DT104 strains were also tested for plasmid profile and for spvC gene indicating the presence of the large serotype specific plasmid (Ssp). No characteristic plasmid profile could be attributed to S. Typhimurium DT104. The serovar-specific large plasmid was detected by PCR for spvC in 100% of DT104 strains and in 77% of the non-DT104 strains. The virulence of two DT104 strains was tested in orally infected day-old chicks and compared with virulence of 4 non-DT104 strains. Higher colonizing virulence of DT104 strains could be established as compared to the other strains.

The cyclohexane tolerance and Phe-Arg-ß-naphtylamide susceptibility of multidrug-resistant Enterobacter cloacae clinical isolates, and the predominance of one PFGE clone in Hungary.

The report concerns the molecular epidemiology, cyclohexane tolerance and Phe-Arg-ß-naphtylamide (PAßN) susceptibility of multidrug-resistant Enterobacter cloacae isolates, with high-level fluoroquinolone resistance collected from healthcare facilities in a nationwide survey. A total of 113 multidrug-resistant E. cloacae isolates (recovered in 1997-2005) were subjected to disc diffusion tests, ERIC-PCR and XbaI PFGE. Representatives of the ERIC-types (n = 67) were tested further with cyclohexane and PAßN, using ciprofloxacin as the substrate. Forty-four per cent of the isolates were derived from the urinary tract, 19% from the bloodstream, 17% from the respiratory tract, and 15% from wound infections. Four ERIC-types (A, B, C and D) were distinguished, but 109 isolates were found to belong to a single, epidemic ERIC type: A. PFGE results suggested that the epidemic-type isolates were of monoclonal origin. Forty-two patients were involved in four outbreaks caused by the epidemic-type strains. Eighty-one cases were found to be nosocomial. At least fourfold reduction in ciprofloxacin MICs was found in the presence of PAßN in 79% of representative isolates (representing types A, C and D); an eightfold or greater reduction in ciprofloxacin MICs in the presence of PAßN (PAßN+) was found in 37% of representative isolates, representing types A and C. Eighty-five per cent of the representative isolates were found to be cyclohexane-tolerant, representing types A, C and D. This is the first report of a wide distribution of cyclohexane-tolerant or PAßN+ strains of E. cloacae. These feature-indicators of adaptive mechanisms that help bacteria to survive in hospital wards may have contributed to the nationwide spread of type A strains.

Dissemination of ST274 Klebsiella pneumoniae epidemic clone in newborn and adult hospital settings harbouring SHV-2A or CTX-M-15 type extended spectrum ß-lactamases-producing known plasmids.

Molecular epidemiology and genetic features of an extended-spectrum ß-lactamase (ESBL) producing Klebsiella pneumoniae epidemic clone (KP-EC) with elevated ciprofloxacin MIC (minimum inhibitory concentration) values from multiple nosocomial outbreaks and sporadic cases between 2006 and 2008 in Hungary were investigated. As a result of continuous monitoring of ESBL-producing KP-ECs, 27 isolates collected from five healthcare facilities were selected for macrorestriction profile analysis by PFGE (pulsed field gel electrophoresis). Of these, 12 strains were isolated from adult inpatients, while 15 strains were from newborns. The MIC values for several antibiotics were determined by agar dilution technique. Molecular typing was further performed by PCR (polymerase chain reaction) and sequencing of several antibiotic resistance genes, plasmid profile analysis, transfer of resistance determinants and multilocus sequence typing (MLST). All isolates showed moderate resistance to ciprofloxacin (MICs ranged from 0.5 to 8 mg L(-1)). PFGE revealed the existence of only one genetic cluster defined as EC IV. PstI digestion of plasmid DNA revealed two highly diverse restriction patterns in "adult" and "newborn" isolates corresponding to plasmids from the Hungarian Epidemic Clone and plasmids isolated from a neonatal nosocomial outbreak in 1998, respectively. Sequence analysis of ß-lactamase genes from plasmids of 14 selected isolates detected bla SHV-2a in strains isolated exclusively from newborns and bla CTX-M-15 in strains isolated exclusively from adult inpatients. MLST established that strains of the PFGE cluster belonged to a novel sequence type ST274. ESBL-producing K. pneumoniae isolates belonging to the novel sequence type ST274 appeared in the newborn and adult hospital settings in Hungary and acquired SHV-2a or CTX-M-15 type enzymes, respectively. Thus, a new antimicrobial resistance strategy for successful conformation to distinct hospital settings was found.

From farm to fork follow-up of thermotolerant campylobacters throughout the broiler production chain and in human cases in a Hungarian county during a ten-months period.

A study tracking thermotolerant campylobacters from the setting of the broilers throughout the whole rearing period, slaughter and sale of chicken products in five consecutive broiler rotations of the same henhouse as well as in two different other farms was conducted in a well-defined geographic area (Hajdú-Bihar county, Hungary) between March 2006 and Feb 2007. All notified cases of human campylobacteriosis in this area during the study period were also included. One hundred and one, 44, 23 and 282 Campylobacter jejuni and 13, 15, 20 and 60C. coli were isolated from broiler houses, slaughterhouses, retail shops and human samples, respectively. Sixty-two isolates collected from broilers or their environment selected from different flocks (57C. jejuni, 5C. coli), 92 isolates collected from abattoirs and retail shops (72C. jejuni, 20C. coli), as well as 85 randomly selected human isolates (74C. jejuni, 11C. coli) were subjected to PFGE analysis using restriction enzymes KpnI and SmaI. Sixty-six of the isolates produced unique Sma-Kpn profiles; the majority (46) of these were of human origin. The remaining isolates formed PFGE clusters of between 2-25 isolates with 14 (12C. jejuni and 2C. coli) main clusters comprised of five or more isolates with identical KpnI-SmaI patterns. Two genetic clones of C. jejuni (clone A, n=25; clone B, n=20) included 18% of isolates from different sources. Generally, isolates from one cluster were found in 1-3 different flocks, notably, clone B was present in three rotations including those from the two independent farms. Six of the seven investigated flocks had one or two characteristic prevalent clones. Transmission of clones between consecutive flocks was frequently seen. Spread of both C. jejuni and C. coli was traced multiple times along the food chain; eight C. jejuni, but no C. coli clones were detected both in broilers and humans. These data suggest that broilers were the major source for C. jejuni but not for C. coli in the studied area and period. For C. jejuni the carryover of strains between consecutive flocks may be a common event, but the strain is eventually replaced by another and consecutive carryover events seem to be infrequent. The majority of the human disease was due to nonepidemic strains; some clones were transmitted from more than one broiler flocks (including epidemiologically unrelated flocks) to humans multiple times.

Molecular characterisation of invasive Streptococcus pyogenes isolates from Hungary obtained in 2004 and 2005.

Our aim was to characterise by molecular techniques group A streptococci isolated from invasive infections in Hungary in 2004-2005. Twenty-six nonduplicate invasive GAS isolates were selected and examined. The mortality rate proved high (52.3%) for those cases (n = 21) where data were available. Predominant emm types were emm1 (n = 13, 50%) and emm80 (n = 5, 19.2%), but other M types (emm4, emm28, emm66, emm81.1, emm82, emm84) were also identified. Eight different PFGE types were distinguished, and each emm type showed an individual PFGE pattern. Our results show that--similarly to results obtained in several other countries--emm type 1 strains predominate among invasive GAS isolates, and that emm 1 type strains recovered from severe streptococcal infections were associated with the presence of the speA gene. The rate for macrolide resistance proved low: only two isolates showed elevated MICs for erythromycin.

Typing of coagulase-negative staphylococci isolated from immunocompromised patients.

A total of 3121 coagulase-negative staphylococcal strains sourced from clinical samples were characterized during a 4-year period. Biotype, antibiotic resistance pattern, phage pattern and slime production was determined. Plasmid profile analysis was performed on related isolates. Thirty percent of strains originated from the Bone Marrow Transplantation Unit, The National Institute of Haematology, Blood Transfusion and Immunology (NIHBTI), Budapest. Staphylococcus epidermidis occurred most frequently (48.8% in total, 58.2% source from NIHBTI). Total bacteriophage typability was 75.9%, and 603 phage patterns were observed. NIHBTI isolates differed in the incidence of multiply resistance, slime positivity and average frequency of phage patterns from the total suggesting spread of a selected hospital population. Statistical analysis of data obtained by typing showed no predominance of any endemic clone: the strains colonizing the immunocompromised patients and isolated from staff and inanimated environment differed from each other in biotype, phage pattern, antibiotic susceptibility, slime production and/or plasmid profile.

Change in phage-sensitivity of Shigella flexneri strains. I. Changes in virulence and phage-sensitivity.

In the course of repeated passaging of virulent Shigella flexneri cultures in liquid media, avirulent variants and variants of reduced virulence appeared. They were easily recognizable by their colonial morphology. The avirulent variants became sensitive to certain phages not lysing the original virulent strains. The change of phage-sensitivity among the variants with no or decreased virulence was significant: chi 2 = 64.25; P less than 0.001. The amplification of phage-sensitivity developed in 15% of the originally avirulent cultures. No association was found between the loss of virulence and a specific phage-sensitivity pattern. In the majority of the cases sensitivity to phage Ms2 developed during the passages, but it was observed in some of the original strains, too. With few exceptions, sensitivity to phage Ms2 was associated with the total or partial degradation of the specific antigen and the antigenic structure detectable by S. flexneri factor sera. These exceptions were cultures with maintained or decreased virulence.

Phenotypic and genotypic properties of methicillin resistant Staphylococcus aureus strains isolated in Hungary, 1997-2000.

An account is given on the activity of the National Center for Phage Typing of Staphylococci in Hungary in the period between 1997 and 2000 related to methicillin resistant Staphylococcus aureus (MRSA) strains originating mainly from hospital infections and sporadic cases. The rate of multiresistant MRSA strains has decreased gradually from 98.1% in 1997 to 74.6% in 2000, accordingly the typability by phages showed a considerable improvement by the international basic phages. Resistance pattern of MRSA strains became narrower in the period of the examinations. With the exception of erythromycin the rate of resistance decreased probably as a consequence of the increased use of erythromycin. The typing method was completed with the phenotypic and genotypic characterization of macrolide resistance. Among 73 MRSA strains type A was the most frequent macrolide resistance group, while type B, C1 and C2 occurred rarely. Type A was frequent also among the few MSSA and CNS strains. Out of the 168 examined S. aureus strains ermA genes occurred in 81.5%; in MSSA and CNS strains ermC1 genes were frequent, both genes are responsible for the target modification. The msrA gene, encoding the increased efflux, occurred only in CNS strains. Comparing the results obtained by phenotyping (phage typing) and genotyping (AP-PCR) methods it is of note that MRSA strains which proved non-typable by phage typing gave suitable results by the AP-PCR.

Phage types and epidemiological significance of Salmonella enteritidis strains in Hungary between 1976 and 1983.

In Hungary, 14 819 human Salmonella enteritidis strains were isolated between 1976 and 1983. Phage type was determined of 10 132 human strains originating from 6852 foci, and of 711 strains isolated from animals and water in this period. The human strains were typable in 99.4% and they belonged into 21 phage types. Five phage types (1, 4, 7, 16 and 17) were more frequent than 1%. Phage type 7 predominated among the strains isolated between 1976 and 1980, including 65.6%-89.3% of the strains. There was a change in the prevalence of phage types from 1980-1981, as phage type 7 was ousted by phage type 1. The date of the change in the predominance of phage types coincided with the considerable increase of S. enteritidis isolates; the number of isolates was nearly fivefold in 1980 of that in 1976. Phage type 7 frequent in the first period proved to be homogeneous; the strains could not be subdivided either by the temperate phages carried by them or by other phages. The incidence of phage types 1 and 7 was nearly the same among the strains derived from animals, food, water and hygienic control examinations, and there was no temporal difference in the frequency of the two phage types as it was observed among the human strains. The human strains originated in 49.5% from outbreaks and in 50.5% from sporadic cases in the country. Of the strains examined for phage type during the eight-year period, 41.9% were isolated from 23 field epidemics, 84 community outbreaks and 757 family infections. Analysing the regional spread of S. enteritidis, the increase in the number of isolates was the highest in counties Tolna, Bács-Kiskun, Somogy and Györ-Sopron. The predominance of phage type 1 was observed in counties Békés, Borsod, Csongrád, Györ-Sopron, Hajdu-Bihar, Pest and Tolna. It was obvious in the case of county Tolna that the source of infection was contaminated egg and baby chicken. Phage type 7 predominated in counties Komárom, Vas and Veszprém. Phage type 4 circulated in counties Csongrád and Pest, phage type 17 in county Fejér and phage type 2 in county Hajdú-Bihar.

Changes in phage-sensitivity of Shigella flexneri strains. II. The F-like character of Ms2-sensitive variants.

Variants sensitive to male specific phage Ms2 appeared among Ms2 non-sensitive Shigella flexneri cultures in the course of liquid medium passages. Sensitivity to Ms2 was lost on acridine orange treatment and was transferable into Ms2 non-sensitive variants. The Ms2 sensitive variants had fimbrial antigen. Electron microscopy showed that Ms2 phages were adsorbed on the F-like fimbriae of these variants. It was assumed that F-like plasmids determining F-like fimbrial antigen were carried by these variants.