Tetracycline resistance in Moraxella catarrhalis clinical strains isolated in Poland.

Moraxella catarrhalis is considered an important, exclusively human respiratory tract pathogen which, along with Streptococcus pneumoniae and Haemophilus influenzae, is classified as one of the most frequent bacterial etiological factors causing upper respiratory tract infections. In this manuscript, we report the existence of five tetracycline-resistant M. catarrhalis strains with confirmed presence of tetracycline resistance tetB gene. The strains were isolated from children under the age of three with signs of upper respiratory tract infections. Our research also investigated the occurrence of virulence genes in these strains and involved the analysis of drug resistance to five antibiotic groups. It is the first description of clinical strains with confirmed presence of drug resistance tetB genes isolated in Europe.

A Novel Approach to Study the Effect of Ciprofloxacin on Biofilms of Corynebacterium spp. Using Confocal Laser Scanning Microscopy.

Non-diphtherial corynebacteria are Gram-positive rods that cause opportunistic infections, what is supported by their ability to produce biofilm on artificial surfaces. In this study, the characteristic of the biofilm produced on vascular and urological catheters was determined using a confocal microscopy for the most frequently involved in infections diphtheroid species. They were represented by the reference strains of Corynebacterium striatum ATCC 6940 and C. amycolatum ATCC 700207. The effect of ciprofloxacin on the biofilm produced by the antibiotic-susceptible C. striatum strain was evaluated using three concentrations of the antimicrobial agent (2 ×, 4 ×, and 6 × the MIC - the Minimum Inhibitory Concentration). The basis for the interpretation of results was the statistical analysis of maximum points readings from the surface comprising a total of 245 areas of the biofilm image under the confocal microscope. It was observed that ciprofloxacin at a concentration equal to 4 × MIC paradoxically caused an enlargement of areas with live bacteria within the biofilm. Biofilm destruction required the application of ciprofloxacin at a concentration higher than 6 × MIC. This suggests that the use of relatively low doses of antimicrobial agents may increase the number of live bacteria within the biofilm, and further facilitate their detachment from the biofilm's structure thus leading to the spread of bacteria into the bloodstream or to the neighboring tissues. The method of biofilm analysis presented here provides the original and novel approach to the investigation of the diphtheroid biofilms and their interaction with antimicrobial agents.Non-diphtherial corynebacteria are Gram-positive rods that cause opportunistic infections, what is supported by their ability to produce biofilm on artificial surfaces. In this study, the characteristic of the biofilm produced on vascular and urological catheters was determined using a confocal microscopy for the most frequently involved in infections diphtheroid species. They were represented by the reference strains of Corynebacterium striatum ATCC 6940 and C. amycolatum ATCC 700207. The effect of ciprofloxacin on the biofilm produced by the antibiotic-susceptible C. striatum strain was evaluated using three concentrations of the antimicrobial agent (2 ×, 4 ×, and 6 × the MIC ­ the Minimum Inhibitory Concentration). The basis for the interpretation of results was the statistical analysis of maximum points readings from the surface comprising a total of 245 areas of the biofilm image under the confocal microscope. It was observed that ciprofloxacin at a concentration equal to 4 × MIC paradoxically caused an enlargement of areas with live bacteria within the biofilm. Biofilm destruction required the application of ciprofloxacin at a concentration higher than 6 × MIC. This suggests that the use of relatively low doses of antimicrobial agents may increase the number of live bacteria within the biofilm, and further facilitate their detachment from the biofilm's structure thus leading to the spread of bacteria into the bloodstream or to the neighboring tissues. The method of biofilm analysis presented here provides the original and novel approach to the investigation of the diphtheroid biofilms and their interaction with antimicrobial agents.

Alternations in DNA gyrase genes in low-level fluoroquinolone-resistant Moraxella catarrhalis strains isolated in Poland.

PURPOSE: The purpose of this study was to investigate the molecular mechanisms of fluoroquinolone resistance in Moraxella catarrhalis clinical strains isolated in Lublin, Poland. MATERIALS AND METHODS: A total of 150 non-duplicate clinical strains of M. catarrhalis were obtained from individuals with signs of upper respiratory tract infection. Bacterial identification was corroborated on the basis of phenotypic and biochemical characteristics as well as with the use of molecular tests. The antimicrobial susceptibility of M. catarrhalis isolates was determined using the disk diffusion method and Etest. Mutations in the gyrase (gyrA and gyrB) and topoisomerase (parC and parE) genes were determined by polymerase chain reaction and sequencing. RESULTS: It was observed that 16.7% of the studied isolates were drug resistant. Resistance to tetracycline was detected for 12% of the strains. Resistance to nalidixic acid, moxifloxacin, and levofloxacin was exhibited by 2.7% of the strains; 1.3% of the strains were resistant to trimethoprim/sulfamethoxazole and 0.7% to erythromycin. Minimum inhibitory concentration values of the four strains demonstrating fluoroquinolone resistance were: 6-12 mg/L for nalidixic acid, 1-1.5 mg/L for levofloxacin, 1 mg/L for moxifloxacin, and 0.25-0.5 mg/L for ciprofloxacin. The research resulted in the detection of mutations in 4 strains, in gyrase gyrA and gyrB genes. In gyrA gene, there occurred mutation G412C as well as four silent transition mutations. Within gyrB gene, there occurred mutation, substitution A1481G, as well as two identical silent mutations. CONCLUSION: Our findings reveal that resistance to fluoroquinolones in M. catarrhalis is connected with amino acid substitutions in gyrA and gyrB genes. To our knowledge, this work is the first description of fluoroquinolone-resistant clinical strains of M. catarrhalis with described mutations in gyrA and gyrB genes isolated in Poland and in Europe.

Human infections caused by free-living amoebae.

[b]Abstract Introduction[/b]. Among free-living amoebae that are widely distributed in nature only four genera/species are known as agents of human infections:[i] Acanthamoeba spp., Naegleriafowleri, Balamuthia mandrillaris[/i] and[i] Sappiniapedata[/i]. These amoebae are not well adapted to parasitism, and could exist in the human environment without the need for a host. Infections due to these amoebae, despite low morbidity, are characterized by relatively high mortality rate and pose serious clinical problems. [b]Objectve[/b]. This review study presents and summarizes current knowledge about infections due to pathogenic and opportunistic free-living amoebae focused on epidemiology, clinical manifestations, diagnosis and treatment based on global literature. [b]State of knowledge[/b]. All four genera have been recognized as etiologic factors of fatal central nervous system infections and other serious diseases in humans. [i]N. fowleri[/i] causes an acute fulminating meningoencephalitis in children and young adults. [i]Acanthamoeba spp[/i]. and [i]B.mandrillaris[/i] are opportunistic pathogens causing granulomatous amoebic encephalitis and disseminated or localized infections which could affect the skin, sinuses, lungs, adrenals and/or bones. [i]Acanthamoeba spp[/i]. is also the main agent of acute eye infection -[i] Acanthamoeba keratitis, [/i]mostly in contact lens wearers. However, there is only one recognized case of encephalitis caused by [i]S. pedata. [/i] [b]Conclusions[/b]. Amoebic diseases are difficult to diagnose which leads to delayed treatment, and result in a high mortality rate. Considering those issues, there is an urgent need to draw more attention to this type of diseases.