Pneumococcal carriage in adults aged 50 years and older in outpatient health care facility during pandemic COVID-19 in Novi Sad, Serbia.

BACKGROUND: Data related to carriage of Streptococcus pneumoniae (Spn) and antimicrobial resistance patterns in middle-aged and older adults are limited. We assessed the carriage of Spn, and its antibiotic resistance patterns, among participants ≥50 years of age living in the city of Novi Sad during the second year of COVID-19 pandemic. METHODS: Analysis of prospectively collected data among participants with or without symptoms of upper respiratory tract infection who visited their elected physicians in the Primary Health Care Centre of Novi Sad (outpatient facility) was conducted from May 18, 2021 to December 7, 2021. Both nasopharyngeal (NP) and oropharyngeal (OP) samples from each participant were collected. RESULTS: A total of 1042 samples from 521 study subjects (1 NP and 1 OP sample from each person) were collected. Sixteen samples from the same number of persons (3.1%, 95% confidence interval: 1.76%-4.94%) were culture positive for the presence of Spn. Overall, the median age of study participants was 71 years (range, 50-93 years; 90th percentile, 77 years), and most (197/521, 37.8%) of them were 70-79 years of age. A majority of the study subjects were: females (324/521; 62.2%), sampled during May and June 2021 (376/521, 72.2%), those who did not have contact with children aged 0-10 years in the family (403/521; 77.4%), without smokers in the household (443/521; 85.0%), and those who did not receive vaccine against Spn (519/521; 99.6%). Out of 16 Spn positive samples, for six participants, Spn carriage serotypes were obtained and there were four vaccine (6A, 11A, 15B, and 18C) serotypes, and two (6C and 35F) non-vaccine serotypes. Remaining 10 (62.50%) samples were non-typeable isolates of pneumococci. Among four vaccine serotypes, two (6A and 18C) were represented in PCV13, and 18C along with the other two (11A and 15B) in PPSV23 vaccine. The highest level of resistance of Spn isolates was observed for erythromycin, (10 or 62.50%), and tetracycline, (7 or 43.75%), one isolate showed resistance to penicillin, ampicillin, and amoxicillin/amoxicillin-clavulanic acid, while none of them were resistant to ceftriaxone, trimethoprim/sulfamethoxazole and levofloxacin. There were three multi-drug resistant isolates; one was identified as 6C (non-vaccine serotype), and two other were non-typeable isolates of Spn. CONCLUSIONS: In this first study conducted in Serbia on Spn carriage in adults ≥50 years of age, we found low prevalence of Spn carriage and identified 6 serotypes of Spn, four of which were represented in vaccines. These results may support future Spn colonization studies among middle-aged and older adults.

Pneumococcal nasopharyngeal carriage in children under 5 years of age at an outpatient healthcare facility in Novi Sad, Serbia during the COVID-19 pandemic.

Objectives: To assess whether pneumococcal nasopharyngeal carriage among children aged 24-60 months reduced during the coronavirus disease 2019 (COVID-19) pandemic in Novi Sad, Serbia, and to investigate the overall prevalence of carriage, serotype distribution and dominant serotypes 2-3 years after the introduction of pneumococcal conjugate vaccine 10. Design and methods: This prospective, observational study was conducted in February-March 2020, September-November 2020 and April-June 2021, enabling the comparison of results in the pre-pandemic/early pandemic period with two periods during the COVID-19 pandemic. Pneumococci were identified by standard microbiological methods. Serotype identification was performed using conventional multiplex polymerase chain reaction assays. Results: Among 1623 children tested, 515 (31.7%, 95% confidence interval 29.4-34.0%) carried pneumococci. A significant increase in prevalence was found between February-March 2020 and September-November 2020 (P=0.0085), with no difference found between September-November 2020 and April-June 2021 (P=0.0524). Pneumococcal colonization was significantly higher in children who were fully vaccinated and among children who attended day care centres. The dominant serotypes were 15B, 6B, 19F, 11A, 6C, 6A, 3, 23F and 19A, representing 66.4% of all isolates. Conclusions: This study found that pneumococcal nasopharyngeal carriage in children aged 24-60 months was high before the COVID-19 pandemic, and then increased during the pandemic. This rules out a major role of COVID-19 in the suppression of carriage and, probably, transmission.

Seroprevalence of antibodies against SARS-CoV-2 virus in Northern Serbia (Vojvodina): A four consecutive sentinel population-based survey study.

BACKGROUND: Monitoring changes of seroprevalence over time is important at the beginning and during of COVID-19 outbreak to anticipate its dynamics and plan an adequate public health response. METHODS: We conducted a repeated cross-sectional investigation among asymptomatic outpatient subjects and covered 0.1% of total population of Northern Serbia (Autonomous Province of Vojvodina). Each participant was tested for anti-SARS-CoV-2 antibodies using an immunochromatographic qualitative test (point-of-care rapid test). In the last round of survey IgG antibodies targeting the S1 subunit of the spike protein and the nucleocapsid protein of SARS-CoV-2 virus were assessed. RESULTS: During the four rounds of survey (between the end of April and the end of September), anti-SARS-CoV-2 seropositivities based on immunochromatographic test results were 2.60% (95% CI 1.80-3.63), 3.93% (95% CI 2.85-5.28), 6.11% (95% CI 4.72-7.77) and 14.60% (95% 12.51-16.89), respectively. After adjusting with results obtained from the Line immunoassay test, the estimated overall seroprevalence increased to 16.67% (95% CI 14.45-19.08) corresponding to 322,033 infections in total by the end of September 2020 in Vojvodina's population. Throughout the course of the study, for every RT-PCR confirmed case of COVID-19, there were 39-87 additional infections in Vojvodina. No significant difference (p>0.05) in SARS-CoV-2 seropositivity regarding gender and between age subgroups was observed over the course of the survey. CONCLUSIONS: The population prevalence of SARS-CoV-2 antibodies implies much more widespread infection in Vojvodina than indicated by the number of confirmed cases. However, our results suggest that the population of Vojvodina has not reached a desirable level of protection from SARS-CoV-2 virus by the end of September 2020.

Endemicity of OXA-23 and OXA-72 in clinical isolates of Acinetobacter baumannii from three neighbouring countries in Southeast Europe.

According to the World Health Organization, bacterium Acinetobacter baumannii is the first on the critical priority list of pathogens in urgent need for new antibiotics. The increasing resistance of A. baumannii to the last-line treatment options, including carbapenems, is a global problem. We report the molecular epidemiology of 12 carbapenem-resistant clinical isolates of A. baumannii collected from hospitalised patients in three neighbouring countries in Southeast Europe: Croatia, Serbia, and Bosnia and Herzegovina, giving an insight into the molecular characterisation and evolutionary history of the acquisition of resistance genes. Besides the blaOXA-23 gene, the endemic presence of OXA-72 oxacillinase of the same origin for more than a decade as the leading mechanism of carbapenem resistance in Southeast Europe was confirmed. To the best of our knowledge, this is the first paper that investigates and analyses the phylogenetic association of the most common mechanisms of resistance to carbapenems in clinical isolates of A. baumannii originating from three neighbouring countries in Southeast Europe.

Aristolochic Acids as Persistent Soil Pollutants: Determination of Risk for Human Exposure and Nephropathy from Plant Uptake.

Exposure to aristolochic acids (AAs) from Aristolochia plants is one of the major global causes of nephropathy, including Balkan endemic nephropathy (BEN); renal failure; and urothelial cancer. The high incidence of BEN on the Balkan Peninsula is assumed to result from consumption of Aristolochia clematitis L. seeds coharvested with crops. Here, we show that AAs are long-lived soil contaminants that enter wheat and maize plants by root uptake with strong pH dependence. Soil and crops from Serbian farms in areas endemic for A. clematitis were found to be extensively contaminated with AAs, with contamination strongly correlated with local incidence of BEN. The persistence of AAs as soil contaminants suggests that weed control for A. clematitis plants is needed to reduce the incidence of BEN and aristolochic acid nephropathy, systematic surveys of soil and crop AA levels would identify high-risk regions, and it is imperative to research soil-remediation methods.

Quantitation of Aristolochic Acids in Corn, Wheat Grain, and Soil Samples Collected in Serbia: Identifying a Novel Exposure Pathway in the Etiology of Balkan Endemic Nephropathy.

While to date investigations provided convincing evidence on the role of aristolochic acids (AAs) in the etiology of Balkan endemic nephropathy (BEN) and upper urothelial cancer (UUC), the exposure pathways by which AAs enter human bodies to cause BEN and UUC remain obscure. The goal of this study is to test the hypothesis that environmental pollution by AAs and root uptake of AAs in the polluted soil may be one of the pathways by which AAs enter the human food chain. The hypothesis driving this study was that the decay of Aristolochia clematitis L., a AA-containing herbaceous plant that is found growing widespread in the endemic regions, could release free AAs to the soil, which could be taken up by food crops growing nearby, thereby transferring this potent human nephrotoxin and carcinogen into their edible parts. Using the highly sensitive and selective high-performance liquid chromatography coupled with fluorescence detection method, we identified and quantitated in this study for the first time AAs in corn, wheat grain, and soil samples collected from the endemic village Kutles in Serbia. Our results provide the first direct evidence that food crops and soil in the Balkans are contaminated with AAs. It is possible that the presence of AAs in edible parts of crops originating from the AA-contaminated soil could be one of the major pathways by which humans become exposed to AAs.

Detection of carbapenemase genes in Klebsiella pneumoniae isolates.

Introduction: Klebsiella pneumoniae is one of the leading causes of serious hospital-acquired infections worldwide among Enterobacteriaceae species. It is the most common producer of carbapenemases in many parts of the world. Objective: The aim of the study was to determine which enzymes were responsible for resistance to carbapenems in Klebsiella pneumoniae strains isolated at the Centre of Microbiology of Public Health Institute of Vojvodina. Methods: A total of 29 Klebsiella pneumoniae non-duplicated strains resistant to at least one carbapenem isolated from clinical samples of hospitalized patients between November 1st 2013 and April 30th 2014 were studied. The species identification and susceptibility were done using VITEK 2 (bioMérieux, Marcy-l'Étoile, France) system. Phenotypic conformation of carbapenemase production was done by double-disc synergy test. PCR technique was performed for detection of genes encoding production of carbapenemases (bls(KPC), bla(VIM), bla (NDM), bla(OXA-48)). Results: Isolates of Klebsiella pneumoniae resistant to at least one carbapenem showed positive on double- disc synergy test between meropenem and dipicolinic acid. All strains positive in phenotypic test contained bla(NDM) gene. In isolates resistant only to ertapenem, neither production of carbapenemases nor presence of genes encoding these enzymes were detected. Among these isolates, nine produced extended-spectrum ß-lactamase. Conclusion: The presence of NDM metallo-ß-lactamase was determined in all Klebsiella pneumoniae isolates resistant to at least one carbapenem.

[Frequency of vancomycin-resistant enterococci isolated from blood cultures from 2008 to 2010].

INTRODUCTION: Enterococci are important hospital-acquired pathogens. The most commonly isolated species of the genus, Enterococcus faecalis and Enterococcus faecium are the third to fourth-most prevalent nosocomial pathogens worldwide. The aim of this study was to determine the frequency of resistance to vancomycin and other antimicrobial agents of Enterococcus spp strains isolated from blood cultures of hospitalized patients. MATERIAL AND METHODS: During the three-year period, from 2008 to 2010, 132 strains of Enterococcus spp isolated from blood cultures of hospitalized patients were tested for their susceptibility to ampicillin, vancomycin, gentamycin (high-level resistance), erythromycin, chloramphenicol, teicoplanin, ciprofloxacin by disc diffusion method according to the Clinical and Laboratory Standards Institute recommendations. Susceptibility of vancomycin resistant E. faecium to the same antibiotics and to linezolid, quinopristin/dalfopristin and tigecyclin was determined using VITEK system. RESULTS AND DISCUSSION: Resistance to vancomycin was detected in 21 (15.9%) Enterococcus spp strains. The percentage of resistance to other antimicrobial agents varied from 23. l% for chloramphenicol to 81.3% for ciproflxacin. All vancomycin resistant enterococci were identified as E. faecium and belonged to phenotype VanA. The resistance to other antibiotics was very high, except for linezolid and quinopristin/dalfopristin (4.7%). The high-level aminoglycoside resistance was 87.6% for gentamycin and 95.2% for streptomycin. All isolates were resistant to ampicillin, teicoplanin and ciprofloxacin. CONCLUSION: The detected high frequency of multidrug-resistant isolates among vancomycin resistant enterococci is of great importance and suggests the need for further monitoring of susceptibility in order to take adequate measures to prevent and control spreading of resistant strains.

[Carbapenems resistance of Acinetobacter spp strains isolated from wound swabs during 2009-2010].

INTRODUCTION: Acinetobacter spp has become an important cause of nosocomial infections due to its great ability to survive and spread in a hospital setting and to develop resistance to many antibiotics. The aim of this study was to examine the resistance to carbapenems and other commonly used antibiotics in strains of Acinetobacter isolated from wound swabs. MATERIAL AND METHODS: In the Laboratories of Microbiology Center at the Institute for Public Health in Novi Sad wound swabs were taken from the patients hospitalized at the Institutes and Departments of the Clinical Center of Vojvodina in Novi Sad. A total of 271 Acinetobacter spp strains were tested to susceptibility on carbapenems and other most commonly used antibiotics by disk diffusion method according to the recommendation of the Clinical and Laboratory Standards Institute. RESULTS AND DISCUSSION: Acinetobacter spp (271 isolates) developed resistance to imipenem and meropenem (67.4% and 64.4%). The resistance to both cephalosporins III and IV generation and ciprofloxacin was 100%. The resistance to ampicillin-sulbactam was 71.4%. CONCLUSION: Our results show high resistance to carbapenems in Acinetobacter spp strains isolated from wound swabs. These facts suggest the need for continuous monitoring of susceptibility in order to take adequate measures to prevent and control spreading of resistant strains.