High Prevalence and Resistance Patterns of Community-Associated Methicillin-Resistant Staphylococcus aureus in the Pomoravlje Region, Serbia

With a view to estimating the prevalence and resistance patterns of CA-MRSA in one region of Serbia, we performed an analysis of MRSA isolates from healthy people and hospitalised patients. The detection of CA-MRSA was carried out by SCCmec typing. In MRSA isolates from hospitalised patients SCCmec types IV and V were found in 76% of the strains. Similar percentage (80%) of CA-MRSA genotypes was present in healthy people. SCCmec type V harbouring MRSA was the most successful clone. Higher prevalence of type V in hospitalised patients to that in healthy people (70% vs 54%) may indicate nosocomial transmissions in at least some hospital units. All MRSA strains from hospitalised patients were resistant to one or more non-β-lactam antibiotics while 52% were multi-resistant. In isolates from healthy people, 16% were sensitive to all non-β-lactam antibiotics and 40% were multi-resistant. Similar percentage of multi-resistant CA- and HA-genotypes occurred in a particular environment (53% vs 50% in hospitalised patients, and 37.5% vs 37.5% in healthy people) indicating selective pressure of antibiotics as a leading force conferring antibiotic resistance. High prevalence of CA-MRSA and high resistance rate both in hospitals and the community suggest that this pathogen has been present in the Pomoravlje Region, central Serbia for years

Nalaz larvi nematoda kod grabljivih vrsta riba

Neamatode iz roda Eustrongylides su paraziti ribojedih ptica. Njihov razvojni ciklus obuhvata dva prelazna domaćina, akvatične oligohete i planktofagne i bentofagne ribe npr. Fundulus ili Gambosa, Neogobius, ali i neke vrste iz porodice Cyprinidae koje kada pojedu inficiranu oligohetu postaju drugi prelazni domaćin. Grabljive vrste riba, kakav je som i smuđ, kada pojedu inficiranu ribu postaju paratenični domaćini dok njih ne pojedu ptice. Ovakvu ulogu mogu imati i vodozemci i gmizavci. Ustanovljeno je da ovi paraziti imaju uspešnu strategiju razvoja, s obzirom da jaje ostaje infektivno i do dve godine u spoljnoj sredini, a u prelaznom domaćinu može biti i preko godinu dana. Čovek nije tipičan domaćin ali se može zaraziti ukoliko jede sirovo ili nedovoljno termički obrađeno meso riba. U ovom radu prikazane su nematode koje se javljaju kod grabljivih slatkovodnih riba u Republici Srbiji. Istraživanja su sprovedena u periodu od 2011-2012 godine na kanalu Dunav-Tisza-Dunav, u gradskom području Novog Sada. Prikupljen je 21 uzorak smuđa (Sander lucioperca) težine od 250-500 g i 52 uzorka soma (Siluris glanis) težine 250-450 g. Urađen je postmortalni pregled abdominalne duplje, digestivnog trakta i drugih visceralnih organa. Utvrđeno je prisustvo nematoda kod 4 jedinke smuđa i 6 jedinki soma, što predstavlja prevalencu od 14.26%, odnosno 11.54%. Larve su bile prisutne u abdomenu, muskulaturi, lumenu želuca i želudačnom zidu gde su paraziti bili inkapsulirani. Broj parazita po ribi kretao se od nekoliko pa sve do 256. Sakupljene nematode fiksirane su u 70% etanol. Nakon fiksacije, svaka nematoda prosvetljivana je u mlečnoj kiselini radi morfološke observacije i identifikacije vrste. Relativni morfometrijski parametri i identifikacija parazita sprovedeni su prema ključevima Bauera (1987), Moraveca (1994) i Andersona (2000). Nematode su identifikovane kao Eustrongylides sp.- larveni oblici. Larve su bile crvenkaste boje. Dužina tela larvi se kretala od 27 – 60.5 mm, a širina od 0.49 – 0.58 mm. Karakteriše ih prisustvo 12 papila na prednjem kraju, respoređenih u dva kruga po 6. Papile unutrašnjeg kruga su nešto izduženije Uzorci za patohistološki pregled uzeti su iz mišića i nodula koji su se nalazili u želudačnom zidu. Uzorci su bojeni standardnom metodom koristeći H&E. Paraziti konzumirani kod sveže i termički slabo obrađene ribe mogu predstavljati značajan rizik za ljude. Sveže meso ribe i tradicionalni riblji proizvodi pre nego što se nađu u prometu moraju biti pregledani na prisustvo nematoda. Adekvatna priprema ribljeg mesa jedna od najvažnijih mera opreza kao i podizanje javne svesti prilikom konzumacije ovih riba

Uticaj sporazuma o stabilizaciji i pridruživanju na uvoz i izvoz ribe i proizvoda od ribe

Sporazum o stabilizaciji i pridruživanju (SSP) je međunarodni ugovor, potpisan 29. aprila 2008. godine između Republike Srbije i Evropske unije (EU). Dve najvažnije obaveze koje Republika Srbija preuzimaovim sporazumom biće uspostavljanje zone slobodne trgovine i usklađivanje zakonodavstva Republike Srbije sa pravom EU. Sporazumom se stvara zona slobodne trgovine između Srbije i EU u prelaznom periodu od šest godina. Rok za liberalizaciju trgovine je određen u skladu sa sposobnošću srpske industrije i poljoprivrede da se prilagode slobodnoj trgovini ali i sa željom Srbije za što bržim završetkom reformi i pristupanjem Evropskoj uniji. Obaveza Srbije se sastoji u postepenom ukidanju carina na uvoz robe poreklom iz Evropske unije u prelaznom periodu. Sa druge strane, Evropska unija ovim ugovorom potvrđuje slobodan pristup robi iz Srbije tržištu Evropske unije. Tempo liberalizacije i stepen zaštite zavisi od stepena osetljivosti proizvoda za industriju Srbije. Definisane su tri grupe indistrijskih proizvoda, prema osetljivosti, za koje će liberalizacija biti ostvarena nakon perioda od dve, pet odnosno šest godina. Za proizvode koji se ne nalaze na ovim listama carine će biti ukinute momentom stupanja na snagu sporazuma. Sporazumom o stabilizaciji i pridruživanju predviđa se i obaveza Republike Srbije da u dogovorenim rokovima uskladi domaće zakonodavstvo sa propisima koji spadaju u pravne tekovine EU. S obzirom na obim pravnih tekovina EU određena su prioritetna područja koja imaju direktan uticaj na stvaranje zone slobodne trgovine između EU i Srbije: zaštita konkurencije i kontrola dodele državnih pomoći (subvencija), pravo intelektualne svojine, javne nabavke, standardizacija i zaštita potrošača. Narodna skupština Republike Srbije 9. septembra 2008. godine ratifikovala je Sporazum o stabilizaciji i pridruživanju (SSP) i Prelazni trgovinski sporazum. SSP i Prelazni sporazum potpisani su 29. aprila 2008.godine u Briselu. Od 1. februara 2009. Srbija jednostrano primenjuje prelazni trgovinski sporazum

Intestinal carriage of vancomycin-resistant Enterococcus spp. among high-risk patients in university hospitals in Serbia: first surveillance report

Background: The screening for intestinal carriage of vancomycin-resistant Enterococcus spp. (VRE) among high risk patients in the Balkan region and molecular epidemiology of VRE is insufficiently investigated, yet it could be of key importance in infection control. The aim of this study was to provide baseline data on VRE intestinal carriage among high-risk patients in Serbian university hospitals, to determine the phenotypic/genotypic profiles of the isolated VRE, to obtain knowledge of local resistance patterns and bridge the gaps in current VRE surveillance. Methods: The VRE reservoir was investigated using stool samples from 268 inpatients. Characterization of isolated VRE stains consisted of BD Phoenix system, genotypic identification, glycopeptide and quinupristin-dalfopristin (Q-D) resistance probing, virulence gene (esp, hyl, efaA, asa1, gelE, cpd) detection and MLVA. Biofilm formation was evaluated by the microtiter plate method. Results: VRE carriage prevalence among at-risk patients was 28.7%. All VRE strains were vanA positive multidrug-resistant Enterococcus faecium (VRfm), harboring ermB-1 (38.9%), esp (84%), efaA (71.2%), hyl (54.5%), asa1 (23.4%), gelE and cpd (11.6%) each. Ability of biofilm production was detected in 20.8%. Genetic relatedness of the isolates revealed 13 clusters, heterogeneous picture and 25 unique MTs profiles. Conclusion: The obtained prevalence of VRE intestinal carriage among high-risk inpatients in Serbia is higher than the European average, with high percentage of multidrug resistance. The emergence of resistance to Q-D is of particular concern. Close monitoring of pattern of resistance and strict adherence to specific guidelines are urgently needed in Serbia

Insight into diversity of bacteria belonging to the order Rickettsiales in 9 arthropods species collected in Serbia.

Rickettsiales bacteria in arthropods play a significant role in both public health and arthropod ecology. However, the extensive genetic diversity of Rickettsiales endosymbionts of arthropods is still to be discovered. In 2016, 515 arthropods belonging to 9 species of four classes (Insecta, Chilopoda, Diplopoda and Arachnida) were collected in Serbia. The presence and genetic diversity of Rickettsiales bacteria were evaluated by characterizing the 16S rRNA (rrs), citrate synthase (gltA) and heat shock protein (groEL) genes. The presence of various Rickettsiales bacteria was identified in the majority of tested arthropod species. The results revealed co-circulation of five recognized Rickettsiales species including Rickettsia, Ehrlichia and Wolbachia, as well as four tentative novel species, including one tentative novel genus named Neowolbachia. These results suggest the remarkable genetic diversity of Rickettsiales bacteria in certain arthropod species in this region. Furthermore, the high prevalence of spotted fever group Rickettsia in Ixodes ricinus ticks highlights the potential public health risk of human Rickettsia infection

Ralstonia pickettii bacteremia in a cardiac surgery patient in Belgrade, Serbia

Ralstonia pickettii is an opportunistic bacterium found in the water environment with an increasing incidence as a nosocomial pathogen. The objectives of this study were to describe R. pickettii bacteremia in a cardiac surgery patient and to evaluate its ability to grow in a saline solution and to form biofilm. The patient in this study underwent mitral and aortic valve replacement surgery with two aortocoronary bypasses. She developed signs of respiratory and renal failure, therefore hemodialysis was started. After 25 days in an intensive care unit, the patient had recurrent episodes of fever with signs of bacteremia. R. pickettii was identified from blood cultures by MALDI-TOF MS. Antimicrobial susceptibility testing was performed using disc diffusion and broth microdilution methods in accordance with EUCAST methodology and results were interpreted following clinical breakpoints for Pseudomonas spp. The isolate was susceptible to all tested antimicrobial agents except aminoglycosides and colistin. Survival of R. pickettii was analyzed in saline solution with four different starting concentrations at 25 degrees C and 37 degrees C for six days. Biofilm capacity was tested using the microtiter plate method. R. pickettii showed substantial growth in saline solution, with starting concentration of 2 CFU ml(-1) reaching 107 CFU ml(-1) after six days. There was no significant difference between growth at 25 degrees C and 37 degrees C. This indicates that storage of contaminated solutions at room temperature can enhance the count of R. pickettii. Our strain did not show the capacity to form biofilm. The patient responded well to adequate treatment with ceftazidime, and after 48 days in ICU she was discharged to convalesce

Evolution and Population Dynamics of Clonal Complex 152 Community-Associated Methicillin-Resistant Staphylococcus aureus

Since the late 1990s, changes in the epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) were recognized with the emergence of community-associated MRSA (CA-MRSA). CA-MRSA belonging to clonal complex 152 (CC152), carrying the small staphylococcal cassette chromosome mec (SCCmec) type V and encoding the Panton-Valentine leukocidin (PVL), has been observed in Europe. The aim of this study was to investigate its origin, evolution, and dissemination. Whole-genome sequencing was performed on a global collection of 149 CC152 isolates spanning 20 years (93 methicillin-susceptible S. aureus [MSSA] and 56 MRSA isolates). Core genome phylogeny, Bayesian inference, in silico resistance analyses, and genomic characterization were applied. Phylogenetic analysis revealed two major distinct clades, one dominated by MSSA and the other populated only by MRSA. The MSSA isolates were predominately from sub-Saharan Africa, whereas MRSA was almost exclusively from Europe. The European MRSA isolates all harbored an SCCmec type V (5C2&5) element, whereas other SCCmec elements were sporadically detected in MRSA from the otherwise MSSA-dominated clade, including SCCmec types IV (2B), V (5C2), and XIII (9A). In total, 93% of the studied CC152 isolates were PVL positive. Bayesian coalescent inference suggests an emergence of the European CC152-MRSA in the 1990s, while the CC152 lineage dates back to the 1970s. The CA-MRSA CC152 clone mimics the European CC80 CA-MRSA lineage by its emergence from a PVL-positive MSSA ancestor from North Africa or Europe. The CC152 lineage has acquired SCCmec several times, but acquisition of SCCmec type V (5C2&5) seems associated with expansion of MRSA CC152 in Europe. IMPORTANCE Understanding the evolution of CA-MRSA is important in light of the increasing importance of this reservoir in the dissemination of MRSA. Here, we highlight the story of the CA-MRSA CC152 lineage using whole-genome sequencing on an international collection of CC152. We show that the evolution of this lineage is novel and that antibiotic usage may have the potential to select for the phage-encoded Panton-Valentine leukocidin. The diversity of the strains correlated highly to geography, with higher level of resistance observed among the European MRSA isolates. The mobility of the SCCmec element is mandatory for the emergence of novel MRSA lineages, and we show here distinct acquisitions, one of which is linked to the successful clone found throughout Europe today

Origin and Evolution of European Community-Acquired Methicillin- Resistant Staphylococcus aureus

ABSTRACT Community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) was recognized in Europe and worldwide in the late 1990s. Within a decade, several genetically and geographically distinct CA-MRSA lineages carrying the small SCCmec type IV and V genetic elements and the Panton-Valentine leukocidin (PVL) emerged around the world. In Europe, the predominant CA-MRSA strain belongs to clonal complex 80 (CC80) and is resistant to kanamycin/amikacin and fusidic acid. CC80 was first reported in 1993 but was relatively rare until the late 1990s. It has since been identified throughout North Africa, the Middle East, and Europe, with recent sporadic reports in sub-Saharan Africa. While strongly associated with skin and soft tissue infections, it is rarely found among asymptomatic carriers. Methicillin-sensitive S. aureus (MSSA) CC80 strains are extremely rare except in sub-Saharan Africa. In the current study, we applied whole-genome sequencing to a global collection of both MSSA and MRSA CC80 isolates. Phylogenetic analyses strongly suggest that the European epidemic CA-MRSA lineage is derived from a PVL-positive MSSA ancestor from sub-Saharan Africa. Moreover, the tree topology suggests a single acquisition of both the SCCmec element and a plasmid encoding the fusidic acid resistance determinant. Four canonical SNPs distinguish the derived CA-MRSA lineage and include a nonsynonymous mutation in accessory gene regulator C (agrC). These changes were associated with a star-like expansion into Europe, the Middle East, and North Africa in the early 1990s, including multiple cases of cross-continent imports likely driven by human migrations. IMPORTANCE With increasing levels of CA-MRSA reported from most parts of the Western world, there is a great interest in understanding the origin and factors associated with the emergence of these epidemic lineages. To trace the origin, evolution, and dissemination pattern of the European CA-MRSA clone (CC80), we sequenced a global collection of strains of the S. aureus CC80 lineage. Our study determined that a single descendant of a PVL-positive methicillin-sensitive ancestor circulating in sub-Saharan Africa rose to become the dominant CA-MRSA clone in Europe, the Middle East, and North Africa. In the transition from a methicillin-susceptible lineage to a successful CA-MRSA clone, it simultaneously became resistant to fusidic acid, a widely used antibiotic for skin and soft tissue infections, thus demonstrating the importance of antibiotic selection in the success of this clone. This finding furthermore highlights the significance of horizontal gene acquisitions and underscores the combined importance of these factors for the success of CA-MRSA