Incidence and antibiotic susceptibility of genital mycoplasmas in sexually active individuals in Hungary.

The aim of this study was to examine the incidence and antibiotic sensitivity of Ureaplasma urealyticum and Mycoplasma hominis strains cultured from the genital discharges of sexually active individuals who attended our STD outpatient service. Samples were taken with universal swab (Biolab®, Budapest, Hungary) into the Urea-Myco DUO kit (Bio-Rad®, Budapest, Hungary) and incubated in ambient air for 48 h at 37 °C. The determination of antibiotic sensitivity was performed in U9 and arginin broth using the SIR Mycoplasma kit (Bio-Rad®, Budapest, Hungary) under the same conditions. Between 01.05.2008 and 31.12.2011, 373/4,466 (8.35 %) genito-urethral samples with U. urealyticum and 41/4,466 (0.91 %) genito-urethral samples with M. hominis infection were diagnosed in sexually active individuals in the National STD Center, Semmelweis University. U. urealyticum was isolated in 12.54 % in the cervix and 4.1 % in the male urethra, while M. hominis was isolated in 1.33 % in the cervix and 0.51 % in the male urethra. The affected age group was between 21 and 60 years old. U. urealyticum strains were sensitive to tetracycline (95.9 %), doxycycline (97.32 %), and azithromycin (85.79 %), and resistant to erythromycin (81.23 %), clindamycin (75.06 %), and ofloxacin (25.2 %). Cross-resistance occurred in 38.71 % of patients to erythromycin and clindamycin. M. hominis strains were sensitive to clindamycin, ofloxacin, and doxycycline in more than 95 %, to tetracycline in 82.92 %, and no cross-resistance was detected among the antibiotics. Our study confirms that the continuously changing antibiotic resistance of ureaplasmas and mycoplasmas should be followed at least in a few centers in every country, so as to determine the best local therapy options for sexually transmitted infection (STI) patients.

Significance of methicillin-teicoplanin resistant Staphylococcus haemolyticus in bloodstream infections in patients of the Semmelweis University hospitals in Hungary.

The purpose of this study was to quantify the impact of Staphylococcus haemolyticus in the epidemiology of the blood stream infection (BSI) and to characterize the rates and quantitative levels of resistance to antistaphylococcal drugs. During an eight-year period, 2967 BSIs of the patients hospitalized in different clinical departments of the Semmelweis University, Budapest, Hungary were analyzed. One hundred eighty-four were caused by S. haemolyticus, amounting to 6% of all infections. The antibacterial resistance of S. haemolyticus isolates was investigated by the broth microdilution method, vancomycin agar screen, population analysis profile and PCR for mecA, vanA and vanB genes detection. Epidemiological investigation was processed by determining phenotypic antibiotic resistance patterns and PFGE profiles. Extremely high MIC levels of resistance were obtained to oxacillin, erythromycin, clindamycin, gentamicin and ciprofloxacin. The incidence of teicoplanin reduced susceptibility revealed 32% without possessing either the vanA or vanB gene by the strains. PFGE revealed 56 well-defined genotypes indicating no clonal relationship of the strains. The propensity of S. haemolyticus to acquire resistance and its pathogenic potential in immunocompromised patients, especially among preterm neonates, emphasise the importance of species level identification of coagulase-negative staphylococci and routinely determine the MIC of proper antibacterial agents for these isolates.

Characterisation of verotoxin-producing Escherichia coli strains isolated from human patients in Hungary over a 7-year period.

The purpose of this study was to characterise verotoxigenic Escherichia coli (VTEC) strains isolated in Hungary from 2000 to 2006. Altogether, 33 human VTEC strains were investigated to define the O:H antigens, verotoxin 1, 2 (vtx1 and 2), intimin (eae), enteroaggregative heat-stable toxin (ast1), autoagglutinating adhesin (saa) and enterohaemolysin (ehlyA) genes and sensitivity to 11 antimicrobial agents. The strains belonged to 14 different O:H serotypes, among which O157:NM (non-motile) was the most prevalent (45%, 15/33). Patients infected with O157 more often presented bloody diarrhoea or haemorrhagic colitis (63%, 12/19) than those infected with non-O157 (46%, 6/14). Haemolytic uraemic syndrome evolved in two patients infected with O26:H11. The vtx1vtx2c toxin gene combination was found in 58% (11/19) and vtx2c alone in 31% (6/19) of the O157 strains. All of the O157 strains possessed gamma1, while two O26 strains had the beta1 intimin gene. Twenty strains (75%, 25/33) carried the ehlyA gene and five non-O157 strains had ast1. The majority of the strains (76%) were resistant to at least one antimicrobial agent, but none of them showed the extended-spectrum beta-lactamase (ESBL) phenotype.

In vitro efficacy of amphotericin B, 5-fluorocytosine, fluconazole, voriconazole and posaconazole against Candida dubliniensis isolates using time-kill methodology.

Candida dubliniensis is a recently described yeast that causes infections in mucosal surfaces as well as sterile body sites. Candida dubliniensis develops resistance to fluconazole (FLC) more rapidly than the closely related species C. albicans. The killing activity of amphotericin B (AMB), 5-fluorocytosine (5FC), FLC, voriconazole (VRC) and posaconazole (POS) was determined against six C. dubliniensis clinical isolates, identified using molecular biological methods and C. dubliniensis CD36 reference strain. Minimum inhibitory concentrations (MICs) were determined using the Clinical and Laboratory Standards Institute standard procedure. Time-kill assays were performed using RPMI-1640 as test media over a 48-h period. AMB proved to be fungicidal at >or=0.5 microg ml(-1) against all clinical isolates after 48 h. 5FC was only fungicidal at 32-64x MIC (4-8 microg ml(-1)) against all C. dubliniensis isolates. FLC, VRC and POS were fungistatic; decrease in colony number was observed only at the highest concentrations tested (8, 4 and 4 microg ml(-1), respectively). Triazoles invariably showed fungistatic effect at concentrations attainable in the serum. In clinical situations when a fungicidal antifungal is desirable, AMB may be used.

Phenotypic and genetic characterisation of methicillin-resistant Staphylococcus aureus strains isolated from the university hospitals of Debrecen.

The purpose of this study was to characterise methicillin-resistant Staphylococcus aureus (MRSA) strains isolated in 2005 at the university hospitals of Debrecen, Hungary. Three hundred and thirty-nine MRSA strains were isolated from 102 patients at 18 different clinics. Their sensitivity to oxacillin and ten other antibiotics was determined. For genotypic analysis, phage typing and pulsed-field gel electrophoresis (PFGE) was performed. The rate of MRSA strains increased to 7.2% in 2005, especially at the clinics of surgery, pulmonology and paediatrics. No vancomycin- or teicoplanin-resistant strains were found. The resistance to erythromycin, clindamycin and ciprofloxacin was nearly 100% and multi-resistance was very frequent. Fifty-eight percent of the isolates belonged to mixed phage types and 8% was non-typable. One PFGE clone contained 58.2% of all strains and two further major clones were found at a separately located clinical block, indicating intra-hospital spread. We can conclude that MRSA exhibits an increasing nosocomial problem also in Hungary.

Evaluation of the new Micronaut-Candida system compared to the API ID32C method for yeast identification.

A new system, Micronaut-Candida, was compared to API ID32C to identify 264 yeast (Candida albicans, C. parapsilosis, C. tropicalis, C. krusei, C. inconspicua, C. norvegensis, C. lusitaniae, C. guilliermondii, C. dubliniensis, C. pulcherrima, C. famata, C. rugosa, C. glabrata, C. kefyr, C. lipolytica, C. catenulata, C. neoformans, Geotrichum and Trichosporon species, Rhodotorula glutinis, and Saccharomyces cerevisiae) clinical isolates. Results were in concordance in 244 cases. Eighteen out of the 20 of discordant results were correctly identified by Micronaut-Candida but not by API ID32C, as confirmed by PCR ribotyping.

Is MRSA more virulent than MSSA?

Numerous clinical studies have indicated, based on mortality rates, that methicillin-resistant Staphylococcus aureus (MRSA) strains are more virulent than methicillin-susceptible S. aureus (MSSA) strains. In contrast, quantitative laboratory examinations of the presence and magnitude of pathogenic mechanisms and virulence factors in strains of MRSA and MSSA have generated conflicting data. The most important reason for these conflicting results is probably the heterogeneic nature of the resistant population. A comparison of selected and congenic MRSA and MSSA sub-populations of the same strain is required to resolve this issue.

The first steps towards fluoroquinolone resistance in Hungarian pneumococci.

The incidence of fluoroquinolone resistance among Hungarian routine laboratory Streptococcus pneumoniae isolates, collected in 2000-2002, in common with other European countries, was very low; only 5/304 strains (1.64%) were resistant to ciprofloxacin (MIC = 4 microg/ml), and the other fluoroquinolones showed full efficacy. However, we could identify the Lys-137-Asp amino acid change, caused by a point mutation in the QRDR of the parC gene, in five strains. Additionally, we observed a definite shift in the minimum inhibitory concentrations (MICs) of all fluoroquinolones towards higher values throughout the study period. These two findings, coupled with the increasing consumption figures of fluoroquinolones, suggest that pneumococcal resistance looks poised to develop in Hungary.

The relationship between serotypes and PFGE genotypes in isolates of Streptococcus pneumoniae from Hungary.

The relatedness of 112 penicillin-non-susceptible isolates of Streptococcus pneumoniae from Hungary was determined by pulsed-field gel electrophoresis (PFGE), serotyping and antibiotic susceptibility tests. The differences in PFGE patterns closely mirrored the changes in resistance. Some genotypes comprised multiple serotypes, and the genetic diversity among certain serotypes was considerable. Generally, serotyping alone was insufficient for epidemiological mapping of pneumococcal isolates. There was considerable serotype diversity, but the five most frequent international serotypes (6, 9, 14, 23, 19) were the most prevalent. In addition, the presence of some well-defined resistant international pneumococcal clones in the Hungarian population was identified.

Pathogenicity and virulence of coagulase negative staphylococci in relation to adherence, hydrophobicity, and toxin production in vitro.

AIMS: To study the pathogenicity and virulence characteristics of Staphylococcus epidermidis, Staphylococcus haemolyticus, and Staphylococcus sapro-phyticus. METHODS: BALB/c mice were challenged intraperitoneally with graded doses of three strains belonging to each species. LD50s were measured for each strain. Haemolysin (alpha- and delta-) and enzyme (DNAase, lipase, and esterase) production in vitro were measured qualitatively and quantitatively. Adhesion to plastic was measured and related to cell surface hydrophobicity among the strains. RESULTS: S saprophyticus proved the most virulent (LD50 = 2.7-2.9 x 10(7) cfu/g body weight) while S epidermidis was the least virulent (LD50 = 6-8 x 10(7) cfu/g body weight). An enlarged spleen was the most common macroscopic pathological feature. Kidney, liver, and more rarely peritoneal abscesses were also seen in the infected animals. No direct correlation was found between adherence in vitro, cell surface hydrophobicity, or toxin/enzyme biosynthesis and virulence in mice. CONCLUSION: The results show that coagulase negative staphylococci are pathogenic in BALB/c mice. It is clear that these bacteria can cause invasive disease. However, the in vitro characteristics of coagulase negative staphylococci are not related to the pathogenicity of the organisms in mice.

A novel labeling procedure of anteiso-fatty acid-containing lipids in staphylococci for investigating the effect of penicillin on lipid release.

L-[4,5-3H]isoleucine was introduced to label anteiso-fatty acid (AIFA)-containing lipids in Staphylococcus aureus SG 511. After an overnight incubation in peptone broth in the presence of 37 kBq L-[4,5-3H]isoleucine/ml, 8.5-13% of the total radioactivity applied was found to be incorporated into the cells. 22.4-25.6% of the incorporated radioactivity was found in AIFA-containing lipids extracted by chloroform-methanol-water (2:1:0.2, v/v/v) at pH 2. The interphase contained 70-75% of the incorporated radioactivity. Lipoteichoic acid, extracted by phenol-water (80:20, w/v) contained less than 1% of the incorporated radioactivity, as measured after purification by hydrophobic interaction chromatography on octyl sepharose gel. Within 1 h after addition of 10 micrograms/ml penicillin G to exponentially growing cultures of S. aureus, that led to non-lytic death of the cells, 11.9-18.1% of the incorporated L-[4,5-3H]isoleucine label were released. Lipids containing AIFA were excreted to 5.4-8.4% of total incorporated activity; this amount represents more than 1/4 of the labeled cellular lipids.

Opsonic requirements and surface hydrophobicity of novobiocin-resistant, coagulase-negative staphylococci.

The opsonic requirement for phagocytosis and killing and cell-surface hydrophobicity of five strains of Staphylococcus saprophyticus isolated from clinical sources were studied. Phagocytosis and killing of bacteria by human granulocytes were measured in suspension. Bacterial aggregating cell-surface hydrophobicity was determined by salt aggregation, and the absorptive hydrophobicity was measured by hydrophobic interaction chromatography. All strains were well opsonised by pooled normal human serum 10%. Ingestion of these bacteria could be detected to a variable extent in the absence of extracellular opsonins; heat-inactivated serum 10% or intravenous IgG concentrate 1 mg/ml improved phagocytosis of all strains. Significantly increased rates of both the ingestion and killing of one of the five strains occurred in the presence of IgG or in the absence of opsonins, compared to those found with each of the other four. This particular strain had significantly stronger adsorptive surface hydrophobicity than the other four strains, and with all strains there was a correlation between hydrophobicity and phagocytosis by granulocytes in the absence of opsonins.

beta-Lactam susceptibility patterns and investigation of cephalosporin hydrolysing beta-lactamases of Gram-negative extraintestinal clinical isolates.

Of more than 3500 isolates of enterobacteriaceae, 48-69% were resistant to aminopenicillins and 11-45% to amoxycillin+clavulanic acid. Resistance to second and third generation cephalosporins was present in 11-17 and 3-8% of Escherichia coli, 47-56 and 15-52% of Klebsiella-Enterobacter, 36-57 and 16-27% of Proteus, Providencia and Morganella isolates. Pseudomonas aeruginosa strains varied in their resistance to antipseudomonal beta-lactams. Isoelectric points, inhibitor profiles and substrate profiles of beta-lactamases extracted from representatives of the resistant strains indicated that the resistance was mainly due to the hyperproduction of chromosomally encoded AmpC beta-lactamases. This was confirmed by plasmid profile and PCR investigations. Extended-spectrum beta-lactamase and metallo-penicillinase producing strains were not found. One Pseudomonas maltophilia strain produced an oxacillinase.

Planning of empirical antibiotic therapy for women with pelvic inflammatory diseases: a geographical area-specific study.

OBJECTIVE: Elaboration of an empiric antibiotic regimen for women with pelvic inflammatory disease (PID) for a geographical area in eastern Hungary. STUDY DESIGN: Pathogens were identified by culturing or polymerase chain reaction (PCR) from 2215 patients with suspected PID between 1 January 1999 and 31 December 2001. Empiric guidelines for PID treatment were based on susceptibility testing of the recovered bacteria, patient acceptance and cost-effectiveness of drugs and recommendations of earlier studies. RESULTS: Chlamydia trachomatis was detected in 11%, Neisseria gonorrhoeae in 2%, Streptococcus spp. in 17%, Enterococcus spp. 9%, genital mycoplasmas in 25%, all obligate anaerobic pathogens in 30% of the patients. All antibiotics chosen for our regimen were effective in vitro against one or more recovered pathogens at least in 80%; this regimen produced 98% clinical cure rate in mild cases of PID. CONCLUSION: Early detection and prompt empirical antimicrobial therapy adapted to the local microflora and its resistance pattern can lead to good clinical results.

Effects of alkali metal ions on some virulence traits of Candida albicans.

The effects of the alkali metal ions (Li+, Na+ and K+) on the growth and on certain virulence factors (adhesion, cell-surface hydrophobicity and germinating ability) of Candida albicans were determined. High concentrations of these ions displayed an inhibitory effect on the growth of the Candida cells; preincubation in their presence showed a negative effect on all virulence factors studied. The changes induced during the preincubation remained there even when high concentration of the ions was removed from the cell suspension. In contrast, a considerable growth was found at high Na+ and K+ concentrations. Although alkali metal ions significantly decreased certain virulence traits of the fungus they did not totally inhibit adhesion and germ-tube formation. This suggests that C. albicans may represent a health hazard even at a high salt concentration.

Prevalence of intestinal parasites in dogs in some urban and rural areas of Hungary.

A total of 490 canine faecal specimens collected in the eastern and northern regions of Hungary were examined for helminth eggs. From the results it appears that more than 50% of the dogs were infected with at least one parasite species. The prevalence of eggs (%) in the two regions was as follows: Toxocara canis (24.3-30.1); Trichuris vulpis (20.4-23.3); Ancylostomatidae (8.1-13.1); Capillaria spp. (0-7.3); Toxascaris leonina (2.1-0); Taenia-type (2.8-2.4); Dipylidium caninum (0.4-1); coccidia (3.5-3.4). Of the positive dogs, 8.5-18.1% harboured two or more species of parasites. The prevalence of parasitic infection was also evaluated according to the maintenance, feeding, and age of the animals. The significance of zoonotic diseases (echinococcosis, toxocarosis, ancylostomatidosis) caused by intestinal helminths makes it necessary to know the infection status of domestic dogs and to take measures for control.

[Epidemiology and public health consequences of human toxocariasis as a frequently occurring urban zoonosis]. / A human toxocarosis mint egy gyakori urbanizált zoonosis epidemiológiája és közegészségügyi kihatásai.

The authors give a review mainly about that worm species of the endoparasites of dogs and cats which are more frequently dangerous for the people living in urban circumstances because of the dissemination faeces of these animals. The life cycle of the nematode Toxocara species in dogs and cats as the final hosts, in inadequate++ hosts like man and the consequences of the zoonoses caused by these worms are briefly discussed. The authors survey the epidemiology, clinical appearances and the prevalence of human infections according to the data of the Hungarian and foreign literature. The clinical symptoms and the danger of this parasitosis running with the increasing number of the pet animals are pointed out. It is emphasized that the wide-range explanatory work and the role of the owners of pets in the decrease of the risks are very important, moreover that organizing the struggle against zoonoses such as toxocarosis is mainly veterinary task, however, its control is common medical and veterinary interest. The recognition, identification, diagnosis, treatment and prevention of human infections can only be effective with the proper knowledge in this zoonosis. Toxocarosis is one of the most frequent helminthozoonoses of townspeople in Hungary.

[Distribution and antibiotic resistance of aerobic bacteria isolated from infected wounds caused by burns]. / Egési sebfertözésekböl izolált aerob baktériumok megoszlása és antibiotikum rezisztenciája.

Distribution according to species and genus and resistance to antibacterial agents was studied with 420 bacterial isolates cultured from the wound excretion of 282 burned-infected patients. Of the isolates 68.2% was Gram-positive and 31.8% Gram-negative. This latter high rate may be due to fecal infection as 21% of the burned patients was between the age of 0-4 years. Of the total isolates 38.3% was Staphylococcus aureus, 16.7% coagulase-negative staphylococcus, 10.7% Pseudomonas strain, 6.9% Escherichia coli and 4.8% Klebsiella. Vancomycin resistant Staphylococcus strain was not found. On the other hand 30-35% of the strains was cross-resistant to methicillin- oxacillin-cefuroxim-clindamycin though these agents are the most potent following the vancomycine. Ceftazidime is the most effective agent for Pseudomonas strains being followed by amikacin, carbenicillin, tobramycin and ceftriaxon. Other Gram-negative bacteria showed strongest sensitivity to ceftazidime and ceftriaxone and these are followed by amikacin.

[Pathogenic properties of coagulase-negative staphylococci in experimental infections]. / Koaguláz-negatív staphylococcusok patogenetikai tulajdonságai experimentalis fertözésekben.

The frequency of persistence of three Staphylococcus epidermidis, Staphylococcus haemolyticus and Staphylococcus saprophyticus strains, respectively, was studied in BALB/c mice at the 10th day of intraperitoneal (ip) challenge. 245 out of 416 mice survived after infections with four bacterial suspensions of different colony forming units (CFU) of each strain. Staphylococci persisted in 61 mice (24,9%). The main sites of persistence were the kidneys, while cocci were rarely isolated from the spleen and the liver. S. epidermidis persisted with a significantly higher rate than the other two species, because S. epidermidis in 28,8%, S. haemolyticus in 4,9%, and S. saprophyticus in 3,6% were reisolated from the organs of the respective infected and surviving animals. The organ persistence was proportional to the amount of bacteria injected. The persistence resulted in subacute microabscesses in the organs. Reisolates of persisting bacteria remained stable in phenotype and genotype concerning antibiotic resistance patterns and biochemical activities for the taxonomic implication, whereas cell surface properties characterizable with phage types altered considerably during persistence. It is concluded that cocci of all three Staphylococcus species are invasive and can persist to a certain extent in the organs of animals with normal immune system, too, after artificial inoculation into the peritoneum i. e. to the serosal surfaces.

[Immunosuppressive effect of cyclophosphamide in experimental coagulase-negative staphylococcal infection]. / Cyclophosphamid immunszuppresszió hatása experimentális koaguláz-negatív staphylococcus fertözésekre.

BALB/c mice were given 200 mg/kg cyclophosphamide intraperitoneally. Three days later, mice were infected intraperitoneally with Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus saprophyticus at 4 inocula ranging between 2 x and 20 x 10(8) CFU/ml suspended in dextran microcarrier solution. Controls were treated only with bacteria. Lethality rates and organ persistence of cocci were significantly higher, and more peritoneal abscesses and adhesions developed in the mice pretreated with cyclophosphamide than in the untreated controls, regardless of the species of Staphylococcus injected. Splenomegaly was also more pronounced indicating a probably enhanced compensatory reactivity of the immune system liberated from suppression 13 days after the administration of cyclophosphamide. Our results show that cyclophosphamide treatment increases the susceptibility of mice to infection by coagulase-negative staphylococci and it is also responsible for a more severe course of the diseases provoked.