Ivan Literak

CTX-M-15-producing Escherichia coli clone B2-O25b-ST131 and Klebsiella spp. isolates in municipal wastewater treatment plant effluents

OBJECTIVESnThe global occurrence of antibiotic resistance genes in bacteria in water environments is an increasing concern. Treated wastewater was sampled daily over a 45 day period from the outflow of a municipal wastewater treatment plant in Brno, Czech Republic, and examined for extended-spectrum β-lactamase (ESBL)-producing bacteria.nnnMETHODSnWater samples were cultivated on MacConkey agar with cefotaxime (2 mg/L) and individual colonies were examined for ESBL production. Phenotypic ESBL-positive bacteria identified as Escherichia coli or Klebsiella spp. were tested for the presence of antibiotic resistance genes, the virulence gene afa/dra and the bla(CTX-M) upstream region. Genetic relatedness was analysed by PFGE, multilocus sequence typing and plasmid analysis.nnnRESULTSnA total of 68 ESBL-producing Enterobacteriaceae isolates were detected in 34 out of 45 wastewater samples. ESBL-producing isolates included 26 E. coli isolates, 4 Klebsiella pneumoniae isolates and 1 Klebsiella oxytoca isolate. The pandemic and multiresistant B2-O25b-ST131 clone was predominant, being detected among 19 E. coli isolates, and 17 of the B2-O25b-ST131 isolates were positive for the FIA replicon and the afa/dra operon and had an IS26 element flanking bla(CTX-M-15). Seventeen of the B2-O25b-ST131 isolates showed closely related PFGE profiles (defined by 84% band similarity) and belonged to identical clonal groups.nnnCONCLUSIONSnThe results highlight the inadequacy of the treatment process in removing multiresistant bacteria from municipal wastewater and point to a risk of transmission of clinically important multiresistant strains, such as the pandemic ST131 clone, to the environment. This is the first study demonstrating the pandemic ST131 clone in wastewater.

Plasmids carrying blaCTX-M-1 and qnr genes in Escherichia coli isolates from an equine clinic and a horseback riding centre

OBJECTIVESnThe aim of this study was to determine the occurrence of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli at an equine clinic and a horseback riding centre, and to discuss the impact of antimicrobial treatment on resistance selection.nnnMETHODSnFaeces from horses, environmental smears and flies were sampled at both the clinic and riding centre. Staff at the equine clinic were also examined. The samples were cultivated on MacConkey agar with cefotaxime (2 mg/L) to isolate ESBL-producing E. coli. The presence of bla and qnr genes was tested by PCR, and transferability was determined by conjugation. Replicon typing and restriction analysis of plasmids harbouring ESBL and qnr genes were performed.nnnRESULTSnE. coli with the blaCTX-M-1 gene were isolated from horses, staff, environmental smears and flies at the two sites. E. coli isolates from the equine clinic harboured an IncHI1 conjugative 235-285 kb plasmid containing blaCTX-M-1, catA1, strA, sul2 and tet(B) genes. Some of these were positive for qnrS1 and/or qnrB19, and were located on 40 or 45 kb IncN or IncX1 conjugative plasmids. The gene blaCTX-M-1 in isolates from the riding centre was carried by IncN (30 kb) and IncI1 (85 kb) conjugative plasmids. Horizontal gene transfer seems to be involved in disseminating E. coli with ESBL and qnr genes at the clinic and riding centre.nnnCONCLUSIONSnThe study illustrates that ESBL-producing E. coli, as well as plasmids carrying ESBL genes of clinical interest, can be easily transferred among horses, humans and flies living in close contact.

The Importance of Ixodes arboricola in Transmission of Rickettsia spp., Anaplasma phagocytophilum, and Borrelia burgdorferi Sensu Lato in the Czech Republic, Central Europe

Wild birds are known to be a reservoir of infectious disease agents and disseminatory hosts of ticks. The purpose of this work was to obtain information about the occurrence of rickettsial, anaplasmal, and borrelial infections in some ticks that parasitize wild birds in the Czech Republic. A total of 549 subadult ticks of three species Ixodes arboricola (75.0%), Ixodes ricinus (23.1%), and Haemaphysalis concinna (1.8%) were collected from 20 species of birds (Passeriformes). Rickettsiae were detected in 44.0% larvae and 24.5% nymphs of I. arboricola collected from Parus major, Poecile palustris, and Sitta europaea. Rickettsiae-positive I. ricinus larvae (13.7%) were collected from P. major, Cyanistes caeruleus, and S. europaea, and 2.6% of nymphs from Erithacus rubecula and Prunella modularis. Comparison of sequences of a gltA gene fragment with data available in GenBank identified Rickettsia helvetica, a spotted fever rickettsia associated with human infections, and other Rickettsia spp. Anaplasma phagocytophilum was found only in two I. ricinus nymphs collected from E. rubecula and P. major. Infections with Borrelia burgdorferi sensu lato were recorded in 1.3% larvae of I. arboricola acquired from P. palustris and P. major and in 11.8% larvae and 25.0% nymphs of I. ricinus collected from P. major, P. palustris, C. caeruleus, Acrocephalus schoenobaenus, Turdus merula, Carpodacus erythrinus, Sylvia atricapilla, P. modularis, and Phylloscopus collybita. Reverse-line blot hybridization showed infections with Borrelia garinii and Borrelia valaisiana and mixed infections with these two genospecies. This is the first record of a high rate of rickettsial infection in I. arboricola subadult ticks acquired from birds in the Czech Republic and in central Europe. Our study suggests that I. arboricola, P. major, and P. palustris play important roles in circulating rickettsiae.

Collyriclosis in Central European hirundines

Cutaneous monostome trematode Collyriclum faba (Bremser in Schmalz 1831) is a digenetic flatworm with unknown life cycle. Here, we provide the first compelling evidence that despite low prevalence of the parasite, European hirundines are parasitized by this species. First host record for sand martin (Riparia riparia) and first European host record for barn swallow (Hirundo rustica) is provided. The birds were captured and checked in ten European and Middle Eastern countries, stretching from Ireland to Bahrain, but only samples from Central Europe (Czech Republic, Hungary and Poland) were positive for C. faba. In total, 164,582 sand martins and 100,443 barn swallows were examined, and seven and two birds had cutaneous C. faba cysts. Even though over 40% of the birds captured were juveniles, all but one infected individuals were adults, equally both males and females. The prevalence of the parasite on Central European hirundines were calculated as one in 20,641 for sand martins and one in 4,484 for barn swallows. All the infected birds were captured in close vicinity of water bodies. All the cysts were found close to the vent or on the legs. No adverse effects on its bird hosts were observed.

Antimicrobial Resistance in Fecal Escherichia coli Isolates from Healthy Urban Children of Two Age Groups in Relation to Their Antibiotic Therapy

ABSTRACT The study was performed in the Czech Republic during 2007 to 2009. Of Escherichia coli isolates from 275 children aged 6 weeks, 36% (n = 177) were resistant to 1 to 7 antibiotics. Of isolates from 253 children aged 6 to 17 years, 24% (n = 205) were resistant to 1 to 5 antibiotics. There was no significant difference in the prevalences of antibiotic-resistant E. coli isolates between these groups of children, even though the consumptions of antibiotics were quite different.

Surface-enhanced laser desorption ionization/time-of-flight (SELDI-TOF) mass spectrometry (MS) as a phenotypic method for rapid identification of antibiotic resistance.

Based on experiments with 10 defined strains of Escherichia coli, we present a new method for bacterial phenotyping using SELDI-TOF mass spectrometry. Changes in bacterial protein profiles in the context of the time of cultivation and the antibiotic environment were minimal. Proteom subprofiling may further distinguish between strains with specific susceptibility to antimicrobials. Mass spec-based methods may become common in the future of bacterial pathogen identification in clinical microbiology diagnostics.

Mites of the genus Neharpyrhynchus Fain (Acariformes, Harpirhynchidae) from Neotropical birds

Abstract Three new species of parasitic mites of the genus Neharpyrhynchus Fain (Acariformes, Harpirhynchidae) are described from Neotropical birds: Neharpyrhynchus chlorospingus sp. n. from Chlorospingus pileatus (Passeriformes, Emberizidae) from Costa Rica, Neharpyrhynchus mironovi sp. n. from Dacnys cayana (Passeriformes, Thraupidae) and Neharpyrhynchus tangara sp. n. from Tangara cayana (Thraupidae) both from Brazil. Neharpyrhynchus trochilinus (Fain) is recorded from 3 new host species of the family Trochilidae (Apodiformes), Panterpe insignis and Eugenes fulgens from Costa Rica, and Amazilia lactea from Brazil. Emended diagnosis of the genus and a key to species are provided; all records of Neharpyrhynchus species are summarized.

Mesocestoides litteratus (Batsch, 1786) (Cestoda: Cyclophyllidea: Mesocestoididae) from the red fox: morphological and 18S rDNA characterization of European isolates

SummaryMesocestoides litteratus (Batsch, 1786) (Cestoda: Cyclophyllidea: Mesocestoidae) is a common parasite of the red fox (Vulpes vulpes) and other carnivores across Europe. There has been considerable debate as to the validity of M. litteratus and other closely related, often sympatric species of Mesocestoides. We examine isolates of M. litteratus from red foxes in the Czech Republic, Slovakia and Spain both morphometrically and by characterization of 18S rDNA. Morphometric ranges of all isolates confirmed their identity as M. litteratus and were usually within the range published formerly. The sequences of 18S rDNA of one or two isolates from each country were analysed. The sequences were the same and distinct from all published Mesocestoides 18S sequences with the exception of tetrathyridia from a lizard in the Czech Republic, which was identical to those of M. litteratus.

Cutaneous trematode Collyriclum faba in wild birds in Costa Rica

SummaryAn adult Collyriclum faba was found in a cutaneous cyst at the base of the lower mandible in a blue-gray tanager (Thraupis episcopus, Thraupidae) examined in the Zona Protectora Las Tablas, Costa Rica on 18 August 2010. A total of 1878 birds belonging to 217 species were examined in the country during 2004, 2009, and 2010. C. faba was found for the first time in blue-gray tanager and for the first time in Costa Rica as well.

Microfilariae in birds in the Czech Republic, including a note on adult nematodes Eufilaria delicata in a song thrush Turdus philomelos

Blood samples of more than 1,100 passerineform birds of 40 species were investigated for the occurrence of microfilariae. In the year 2005, 3 out of 677 birds of 31 species (prevalence 0.4%) were infected with microfilariae during the post-nesting period. During the pre-nesting period in the year 2007, 11 out of 438 birds of 31 species were infected with microfilariae (prevalence 2.5%). Both the pre-nesting and post-nesting examinations were conducted at the same location in the northeastern part of the Czech Republic. The microfilariae of the Eufilaria delicata and Ornithofilaria mavis species were found in Turdus merula, Turdus philomelos, and Erithacus rubecula (Passeriformes, Turdidae). Single individual of Poecile montanus (Passeriformes, Paridae) was infected with undetermined microfilariae. The morphometric variability of microfilariae found in T. philomelos, E. rubecula, and Poecile montanus were recorded. Infections caused by microfilariae E. delicata were more frequent than infections caused by O. mavis. Seven adult nematodes E. delicata were found in a subcutaneous cyst on the heel joint in one T. philomelos, which is the first record of adult E. delicata nematodes in birds in the Czech Republic.