The spread of carbapenem-resistant A. baumannii in Lithuanian hospitals is strongly associated with strains belonging to ECII and carrying a GR2 plasmid encoding two blaOXA-72 genes. The genetic environment of pAB120 supports the role of site-specific recombination associated with the acquisition of carbapenem-hydrolysing class D β-lactamases.

In this study, the genetic organization of three novel genomic antibiotic resistance islands (AbaRs) in Acinetobacter baumannii isolates belonging to group of European clone II (EC II) comM integrated sequences of 18-, 21-, and 23-kb resistance islands were determined. These resistance islands carry the backbone of AbaR-type transposon structures, which are composed of the transposition module coding for potential transposition proteins and other genes coding for the intact universal stress protein (uspA), sulfate permease (sul), and proteins of unknown function. The antibiotic resistance genes strA, strB, tetB, and tetR and insertion sequence CR2 element were found to be inserted into the AbaR transposons. GenBank homology searches indicated that they are closely related to the AbaR sequences found integrated in comM in strains of EC II (A. baumannii strains 1656-2 and TCDC-AB0715) and AbaR4 integrated in another location of A. baumannii AB0057 (EC I). All of the AbaRs showed structural similarity to the previously described AbaR4 island and share a 12,008-bp backbone. AbaRs contain Tn1213, Tn2006, and the multiple fragments which could be derived from transposons Tn3, Tn10, Tn21, Tn1000, Tn5393, and Tn6020, the insertion sequences IS26, ISAba1, ISAba14, and ISCR2, and the class 1 integron. Moreover, chromosomal DNA was inserted into distinct regions of the AbaR backbone. Sequence analysis suggested that the AbaR-type transposons have evolved through insertions, deletions, and homologous recombination. AbaR islands, sharing the core structure similar to AbaR4, appeared to be distributed in isolates of EC I and EC II via integration into distinct genomic sites, i.e., pho and comM, respectively.

A total of 456 non-repetitive Escherichia coli isolates from human clinical specimens (urinary, n5134; cervix, vagina and prostate, n552; blood, pus and wounds, n545), healthy animals (cattle, n545; poultry, n520) and diseased animals (cattle, n553; swine, n564; poultry, n543) obtained in Lithuania during the period 2005-2008 were studied for trimethoprim (TMP) resistance and the prevalence of dfr genes. A TMP resistance rate in the range of 18-26 % respective to the origin was found in clinical isolates, 23-40 % in isolates from diseased animals and 9-20 % in isolates from healthy animals. Of 112 TMP-resistant isolates, 103 carried at least one of the six dfrA genes (dfrA1, dfrA5, dfrA8, dfrA12, dfrA14 and dfrA17) as determined by multiplex PCR and RFLP. The dfrA1 and dfrA17 genes were found most frequently in clinical isolates (17 and 19 isolates, respectively), whilst dfrA1 and dfrA14 genes dominated in isolates of animal origin (25 and 13 isolates, respectively). The dfrA5, dfrA12 and dfrA8 genes were detected at lower frequencies. The association with class 1/class 2 integrons was confirmed for 73-100 % of dfr genes found in most groups of isolates, except for the isolates from diseased swine. In this group, the majority of dfr-positive isolates (67 %, 8/12) carried dfrA8 (6/12) or dfrA14 genes (2/12) that were not associated with integrons. Non-integron location was also confirmed for the remaining dfrA8 genes (six clinical isolates and one isolate from diseased cattle) and for dfrA14 genes (two isolates from diseased cattle and swine each). All cassetteindependent dfrA14 genes were found to be located within the strA gene. This study on the prevalence and distribution of TMP resistance genes among E. coli isolates of human and animal origin in Lithuania demonstrates that dfr genes are carried most frequently as gene cassettes within class 1 and/or class 2 integrons. However, TMP resistance in some of the isolates was found to be mediated by non-integron-associated dfrA8 and dfrA14 genes, indicating the existence of alternative sources for the spread of resistance.

The objective of the study was to evaluate the antimicrobial susceptibility of enterococci spread in raw products of poultry origin intended for human consumption in Lithuania. Samples were obtained from retail markets all over the country. Fifty-eight samples (83%) from a total of 70 tested were positive for Enterococcus spp.Enterococcus faecium (36.2%), E. faecalis (29.3%) and E. hirae (17.2%) were the most prevalent species. Susceptibility testing was based on a microdilution test. Results were interpreted according to clinical breakpoints. The most frequent resistance was demonstrated to tetracycline (84.5%), tylosin (64.5%), erythromycin (63.8%) and ciprofloxacin (36.2%). No resistant strains were found to vancomycin, linezolid and tigecycline. Susceptibility to antimicrobials varied among different species of enterococci. E. faecalis demonstrated more frequent resistance to tylosin, E. hirae -to nitrofuranes. E. faecium was more frequently resistant to penicillin and tetracycline. Both E. faecium and E. hirae demonstrated more frequent resistance to fluoroquinolones than E. faecalis. Susceptibility to other antimicrobials statistically was unreliable between these species of enterococci. PRACTICAL APPLICATIONSEnterococci are widely spread in raw poultry products. Antimicrobial resistance is highly important for antimicrobials that are used in human 3 Corresponding Journal of Food Safety 30 (2010) 902-915. 902

Antibiotic-resistant Escherichia coli (n = 191) and Salmonella enterica (n = 87) isolates of human and animal origin obtained in Lithuania during 2005-2008 were characterized for the presence and diversity of class 1 and 2 integrons. E. coli isolates were obtained from patients with urinary tract infections (UTIs) (n = 59) and both healthy and diseased farm animals, including poultry (n = 54), swine (n = 35), and cattle (n = 43). Isolates of non-typhoidal S. enterica were recovered from salmonellosis patients (n = 37) and healthy animals, including poultry (n = 31) and swine (n = 19). The presence of integrons, their gene cassette structure, and genome location were investigated by polymerase chain reaction, restriction fragment-length polymorphism, DNA sequencing, Southern blot hybridization, and conjugation experiments. Forty percent of the E. coli and 11% of the S. enterica isolates carried class 1 integrons, whereas class 2 integrons were found in E. coli isolates (9%) only. The incidence of integrons in human UTIs and cattle isolates was most frequent (p < 0.01). A total of 23 different gene cassettes within 15 different variable regions were observed. Seven different integron types, all of them transferable by conjugation, were common for isolates from human infections and for one or more groups of animal isolates. The most prevalent integron types contained arrays dfrA1-aadA1 (36%), dfrA17-aadA5 (23%), and dfrA1-sat1-aadA1 (78%). Two E. coli isolates from humans with UTIs harbored class 1 integron on conjugative plasmid with the novel array type of 4800 bp/dfrA17-aadA5Δ-IS26-ΔintI1-aadB-aadA1-cmlA residing on the Tn21-like transposon. Three S. enterica isolates from swine contained class 1 integron with the newly observed array type of 1800 bp/aadA7-aadA7. Integrons of 10 different types of both classes were located on transferable plasmids in E. coli and S. enterica. Our study demonstrated the existence of a considerable and common pool of transferable integrons in E. coli and S. enterica present in clinical and livestock environment in Lithuania.

Tigecycline is a semisynthetic analogue of earlier tetracyclines and represents the first member of a novel class of antimicrobials – glycylcyclines – recently approved for clinical use. It is active against a broad range of gram-negative and gram-positive bacterial species including clinically important multidrug-resistant nosocomial and community-acquired bacterial pathogens. The exact molecular basis of tigecycline action is not clear at present, although similarly to the tetracyclines, it has been shown to inhibit the translation elongation step by binding to the ribosome 30S subunit and preventing aminoacylated tRNAs to accommodate in the ribosomal A site. Importantly, tigecycline overcomes the action of ribosomal protection proteins and is not a substrate for tetracycline efflux pumps of most bacteria – well-known and prevalent cellular mechanisms of microbial tetracycline resistance. The present review summarizes current knowledge on the molecular mechanism of the tigecycline action, antibacterial activity against various bacteria, clinical application, development of resistance to glycylcyclines.

ABSTRACT:The aim of this study was to test and analyse the antimicrobial susceptibility of Enterococcus isolates from Lithuanian poultry farms. Investigations were carried out during the years 2008-2009. The sampling sites, located all over the country, included eight poultry farms of large capacity. All samples were collected from broilers. Enterococcus spp. were isolated from intestines immediately after slaughtering. A total of 160 samples were collected, 20 samples from each farm. The MICs (Minimum Inhibitory Concentrations) of eleven antimicrobial agents were determined for each of the isolates using the broth microdilution method with specific microtitre plate panels (Trek Diagnostic Systems, Inc.). Susceptibility according to clinical breakpoints of chloramphenicol, linezolid, erythromycin, penicillin, quinupristin/dalfopristin, tetracycline, vancomycin, ciprofloxacin and nitrofurantoin was evaluated. One hundred and forty seven samples (92%) from a total of 160 tested samples were positive for Enterococcus spp., however, only 74 strains were selected as non-duplicate isolates. The most predominant species were identified as E. faecium (38%), E. faecalis (17.5%), E. gallinarum (12%) and E. casseliflavus (12%). The most frequent resistance properties were resistances to tetracycline (75.6%), erythromycin (56.8%) and ciprofloxacin (41.9%). No strains resistant to vancomycin and linezolid were found. High percentages of susceptibility to chloramphenicol (82.4%) and penicillin (71.6%) were also observed. A high MIC of tigecycline (≥ 1 mg/l) to 12.2% of enterococci was determined during this study. 44.6% of tested strains had a high MIC (≥ 64 mg/l) to tylosin. There was no significant correlation found between resistances of different species to different antimicrobial agents in vitro.

(2009). Nationwide survey of extendedspectrum b-lactamase-producing Enterobacteriaceae in the French community setting.