Aim: During the pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), many countries reported a significant decrease in the prevalence of influenza virus cases. The study aimed to characterize the flu seasons from 2018 to 2023 in Sarajevo Canton, Bosnia and Herzegovina (B&H), and to assess the possible impact of the SARS-CoV-2 pandemic on the influenza A and B virus circulation.  Methods: The CDC Human Influenza Virus Real-Time RT-PCR Diagnostic Panels were used for the detection of influenza virus A and B, and subtyping of influenza virus A (H1pdm09 virus and H3). The data for this regis-try-based retrospective study were collected at the Clinical Centre of the University of Sarajevo, Unit for Clinical Microbiology (the laboratory that acts as a referral for the detection and characterization of influenza virus and SARS-CoV-2 in Federation B&H). Results: In the 2018/2019 and 2019/2020, an equal percentage of positive cases was recorded (148/410; 36%, and 182/504; 36%, respectively). The absence of the influenza virus was observed in 2020/2021. During 2021/2022, influenza virus was detected among 19/104 (18%) patients and slightly increased in 2022/2023 (45/269; 17%). The switch of the influenza B virus lineage was observed. Conclusion: The SARS-CoV-2 virus had an impact on the prevalence of influenza virus infection among the population of the Sarajevo Canton, B&H. Since the interactions between these two viruses are not entirely clear, awareness of a possible threat to public health is crucial.

Introduction. At the end of 2019 and the year before, there was a significant spread of measles in the World Health Organization (WHO) European Region.Gap statement. Among the countries that reported, a measles outbreak was Bosnia and Herzegovina (BiH).Aim. To describe the measles outbreak in BiH (an entity of the Federation of BiH, FBiH) in 2019.Methodology. Confirmatory IgM serology, measles nucleic acid detection by real-time RT-PCR and virus genotyping were done in the WHO-accredited laboratory for measles and rubella at the Clinical Center of the University of Sarajevo, Unit for Clinical Microbiology. Genotype was determined in all measles-RNA-positive cases by sequence analysis of the 450 nt fragment coding the C-terminal of measles virus nucleoprotein (N).Results. From 1 January to 31 December 2019, 1332 measles cases were reported, with the peak observed in April 2019 (413/1332, 31.01 %). Sarajevo Canton had the highest incidence, number of cases and percentage (206.4; 868/1332; 65.17 %) of measles cases. Around four-fifths of infected persons were unvaccinated (1086/1332, 81.53 %), while 4.58 % of the patients (61/1332) were immunized with one dose of measles-containing vaccine. The highest proportion of cases was found in children 0-6 years of age (738/1332, 55.41 %). Measles IgM positivity was determined in 75.88 % (346/456), while virus RNA was detected in 82.46 % (47/57) of the swab samples. All measles virus sequences belonged to genotype B3. SNP (position 216: C=>T) was detected in 1 of the 40 sequences obtained during this outbreak.Conclusion. Due to suboptimal immunization coverage, BiH belongs to countries at a high risk for measles outbreaks. Post-COVID-19 (coronavirus disease 2019) pandemic, targeted and tailored strategies are required to ensure routine vaccination demand and acceptance and broad partner and stakeholder group participation.

We report on an ongoing measles outbreak in the Federation of Bosnia and Herzegovina with 141 cases notified between week 52 2023 and week 6 2024. Among those with known vaccination status, 97% were unvaccinated and the most affected group is children under the age of 5 years (n = 87) who were not vaccinated during the pandemic years. Sixty-eight cases were hospitalised, the most common complications were measles-related pneumonia and diarrhoea. The sequenced measles viruses from four cases belonged to genotype D8.

Introduction. Bosnia and Herzegovina (B and H) has been recognized for decades as a country with a high risk of diseases caused by hantaviruses.Gap statement. The severe acute respiratory syndrome-associated coronavirus 2 (SARS-CoV-2) pandemic has diverted attention from many pathogens, including hantavirus.Aim. To provide a socio-demographic, temporal, geographical and clinical laboratory overview of the expansion of hantavirus infection cases during the SARS-CoV-2 pandemic in B and H in 2021.Methodology. The RecomLine HantaPlus IgG, IgM immuno-line assay (Mikrogen, Germany) was used to detect IgG and IgM antibodies to hantavirus serotypes in human sera from clinically suspected cases.Results. In 2021 (January-October), the number of confirmed cases of hantavirus infection and tested persons (92/140; 65,71 %) was higher than in the previous 2 years, 2020 (2/20; 10.00 %) and 2019 (10/61; 16.39 %). Most of the infected persons were men (84/92; 91.30 %). Hantavirus infections were recorded from January to October 2021, and the peak was reached in July (25/92; 27.17 %). Six out of 10 cantons in the Federation of Bosnia and Herzegovina (FB and H) were affected, namely Sarajevo Canton, Central Bosnia Canton, Neretva Canton, Zenica-Doboj Canton, Posavina Canton and Bosnian-Podrinje Canton Goražde, in descending order. Of the 38/92 (41.30 %) infected patients with characteristic clinical manifestations of haemorrhagic fever, including renal (mainly) or pulmonary syndrome, 32/92 (34.78 %) were hospitalized in the Clinical Center of the University of Sarajevo. Two cases were detected with dual infection, hantavirus (Puumala) with Leptospira in one and SARS-CoV-2 in another case. The largest number of infections was related to Puumala (PUUV) (83/92; 90.22 %), while the rest of the infections were caused by the hantavirus Dobrava serotype (DOBV).Conclusion. The reported infections were probably caused by exposure of individuals to at-risk areas inhabited by contaminated rodents as natural reservoirs of hantavirus. As a highly endemic area, B and H requires continuous monitoring and increased awareness of this problem.

This study evaluated the performance of the COVID-19 Ag-RDT compared to the real-time reverse transcription-polymerase chain reaction (rtRT-PCR) for SARS-CoV-2 detection and its use among patients referred for emergency admission. A total of 120 nasopharyngeal swabs were collected from patients referred for emergency admission and immediately preceded for testing to the Unit of Clinical Microbiology. Out of 60 Ag positive tests, 53 (88.3%) were confirmed by rtRT-PCR, while 7 (11.7%) tested negative (false positives). Out of 60 Ag negative tests, 56 (93.3%) were confirmed negative by rtRT-PCR, and 4 (6.7%) were positive (false negatives). Ct value comparison was performed for 53 samples that were positive by both methods: 8 (15.1%) isolates had Ct value up to 20; 37 (69.8%) 21 to 30 and 8 (15.1%) 31 to 40, respectively. The sensitivity of the analyzed rapid Ag test was 92.9%, and specificity 88.9%. The accuracy of the Ag test was 90.8%. This study has shown that rapid Ag tests can be used in emergency admissions to healthcare facilities. However, rtRT-PCR should be considered after negative antigen test results in symptomatic patients, and after positive antigen test results in asymptomatic persons.

Aim Phlebotominae sandflies are primary vectors of phleboviruses, causing the sandfly fever disease. The aim of this study was to detect and report the presence of flaviviruses in Phlebotominae sandflies captured in Bosnia and Herzegovina. Methods After a microscopic and morphometric analysis, the final identification of collected Phlebotomus specimens was confirmed by PCR, using a hemi-nested polymerase chain reaction on extracted and reversely transcribed RNA. Results We obtained a 155 nt long fragment of the viral non-structural protein 5 (NS5) gene (GenBank accession no. MN090154). The acquired nucleotide sequence, provisionally named as Drežnica, showed a maximum of 70-80% identity in 70-88% (110-137 nucleotides) of the query coverage with several Anopheles, Sabethes, Calbertado and Culex flaviviruses. Maximum likelihood phylogenetic analysis showed that the new flavivirus Drežnica clusters together with the flavivirus isolated from Culiseta annulata mosquitos. Conclusion We report the presence of flavivirus in Phlebotominae sandflies, captured in Drežnica, Herzegovina for the first time. The next phase of research will be directed towards virus cultivation, obtaining a longer or complete virus sequence and clarifying the medical and epidemiological importance of the Drežnica virus.

Klebsiella pneumoniae is the second most prevalent gram-negative rod that causes nosocomial infections in hospitalized or otherwise immunocompromised patients. It can develop multiple drug resistance that results in limited treatment options and increased use of carbapenems. Various mechanisms are related to the development of carbapenem resistance in K. pneumoniae. The aim of this study was to perform phenotypic and molecular characterization of clinical isolates of carbapenemase-producing K. pneumoniae from two outbreaks recorded in 2017 and 2018 in Clinical Center University of Sarajevo, Bosnia and Herzegovina. Identification of K. pneumoniae isolates was carried out on the basis of morphological, cultural, and biochemical characteristics. Interpretation of antimicrobial resistance was performed according to EUCAST breakpoints. There were four different resistotypes of carbapenemase-producing K. pneumoniae in this study and all were confirmed positive for blaOXA-48 carbapenemase. Rep-PCR fingerprinting of these strains showed the presence of the two different genetic patterns with no similarity between them. The monitoring, surveillance, and molecular typing are essential to control the emergence of multidrug-resistant strains in nosocomial settings, and to reduce the frequency of outbreak occurrence.

Aim To present combined measles cases data and phylogenetic analysis of the virus circulated in 2018 in the Federation of Bosnia and Herzegovina (FB&H, the entity of Bosnia and Herzegovina), in order to analyse endemic transmission patterns of circulating strains and its implications for elimination efforts. Methods The data were derived from epidemiological case investigations and laboratory diagnoses based on serology, molecular detection and genotyping of the measles virus. Results During 2018 16 measles cases were reported in FB&H, of which five were classified as laboratory confirmed cases, one was an epidemiologically linked case and 10 were clinically compatible cases. Among them 12 (75.00%) cases were unvaccinated or had unknown vaccination status. The most affected population was up to 14 years of age (13/16; 81.25%). None of the cases was fully vaccinated. Viruses of other genetic lineages had been introduced in FB&H in the recent period. Two virus lineages of genotype B3 were identified. Phylogenetic analysis indicated the presence of a unique sequence of measles B3 virus in FB&H (Sarajevo). Conclusion Further strengthening of measles surveillance system and renewed efforts to increase vaccination levels are necessary to prevent disease and for elimination setting.

Aim This cross-sectional study of a group of women with abnormal cytology and high-risk human papillomavirus (hrHPV) infection compared genotyping HPV DNA and mRNA assays according to two age categories of women (S1: ≤30 and S2: >30 years). Methods The hrHPV DNA positive results of 105 cervical samples of women were pooled and those harbouring HPV-16, 18, 31, 33 and/or 45 DNA were tested for the type specific HPV oncogene E6/E7 overexpression (mRNA). Results Although HPV DNA testing showed a higher proportion of women infected by any of five hrHPVs in S1 group, total agreement of hrHPV DNA and mRNA positive results was higher in S2 group of women (75.8% v. 83.9%). The most prevalent type in both age groups was HPV-16. A 100% agreement of positivity of both tests was noted for HPV-18 and 33 in S1 group, and for HPV-18 in S2 group. Increasing concordance of HPV-16 and 31 DNA and mRNA positive results with the severity of cervical cytology was observed in S1 group of women. Absolute matching (100.0%) of positivity of both diagnostic tests was recorded in S2ASCUS group (for HPV-16, 18 and 33), in S1HSIL (for HPV-16, 18, 31 and 33), in S1LSIL category (for HPV-18 and 33) and in S2HSIL group (for HPV-18). Conclusion The results indicate the possibility of predicting the risk of persistent infection only by HPV DNA typing test, with no need for additional RNA testing in categories of infected women showing a high (absolute) agreement of positivity of both tests.