Between March 5 and July 25, 2020, the total number of SARS-CoV-2 confirmed cases in Bosnia and Herzegovina (BH) was 10 090 corresponding to a cumulative incidence rate of 285.7 per 100 000 population. Demographic and clinical information on all the cases along with exposure and contact information was collected using a standardized case report form. In suspected SARS-CoV-2 cases, respiratory specimens were collected and tested by real-time reverse-transcriptase polymerase chain reaction (rRT-PCR) assay. The dynamic of the outbreak was summarized using epidemiological curves, instantaneous reproduction number Rt and interactive choropleth maps for geographical distribution and spread. The rate of hospitalization was 14.0% (790/5646) in Federation of Bosnia and Herzegovina (FBH) and 6.2% (267/4299) in Republic of Srpska (RS). The death rate was 2.2% (122/5646) in FBH and 3.6% in the RS (155/4299). After the authorities lifted mandatory quarantine restrictions, the basic reproduction number increased from 1.13 on May, the 20th to 1.72 on May the 31st. The outbreak concerns both entities, Federation of Bosnia and Herzegovina and Republika Srpska, and it is more pronounced in those aged 20-44 years. It is important to develop the communication and emergency plan for the SARS-CoV-2 outbreak in BH, including the mechanisms to allow the ongoing notification and updates at the national level.

Aim The damage caused by the COVID-19 pandemic has made the prevention of its further spread at the top of the list of priorities of many governments and state institutions responsible for health and civil protection around the world. This prevention implies an effective system of epidemiological surveillance and the application of timely and effective control measures. This research focuses on the application of techniques for modelling and geovisualization of epidemic data with the aim of simple and fast communication of analytical results via geoportal. Methods The paper describes the approach applied through the project of establishing the epidemiological location-intelligence system for monitoring the effectiveness of control measures in preventing the spread of COVID-19 in Bosnia and Herzegovina. Results Epidemic data were processed and the results related to spatio-temporal analysis of the infection spread were presented by compartmental epidemic model, reproduction number R, epi-curve diagrams as well as choropleth maps for different levels of administrative units. Geovisualization of epidemic data enabled the release of numerous information from described models and indicators, providing easier visual communication of the spread of the disease and better recognition of its trend. Conclusion The approach involves the simultaneous application of epidemic models and epidemic data geovisualization, which allows a simple and rapid evaluation of the epidemic situation and the effects of control measures. This contributes to more informative decision-making related to control measures by suggesting their selective application at the local level.

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.

Aim To identify E. coli from chicken meat, establish their antibiotic resistance profiles and to confirm ESBL isolates with real time PCR, as well as to identify risk factors and farming practice associated with the antimicrobial resistance E. coli. Methods The study included 100 chicken skin samples collected randomly from retail supermarkets, butcheries and slaughterhouses. Disk susceptibility testing was performed using the Kirby-Bauer method. Detection of ESBL-producing isolates was performed with double disk synergy test. Molecular analysis of phenotypic ESBL-producing Escherichia coli strains was performed at 7500 real time PCR System. Molecular-genetic analysis included detection of CTX-M 1, 2, and 9 gene families and mutations in the TEM and SHV encoding extended spectrum β-lactamases. Results Prevalence of the phenotypic ESBL-producing E. coli isolates was 29%, and they exhibited remarkable sensitivity to carbapenems (100%) as well as to amikacin (93.10%). All ESBL-producing strains were multidrug resistant. Molecular analysis was performed as the final confirmation of the production of extended spectrum β - lactamases for 24 isolates out of 29 phenotypicaly ESBL-producing E. coli isolates. Conclusion It is important to pay attention to people's awareness of bacterial antimicrobial resistance in food chain, as well as to understand its effects on human health and the environment. Phenotypic and molecular analysis demonstrated the presence of ESBL-producing E. coli isolates from chicken skin samples.

Whole Genome Sequence of four samples from COVID-19 outbreaks was done in two laboratories in Bosnia and Herzegovina (Veterinary Faculty Sarajevo and Alea Genetic Center). All four BiH sequences cluster mainly with European ones (Italy, Austria, France, Sweden, Cyprus, England). The constructed phylogenetic tree indicates probable multiple independent introduction events. The success of future containment measures concernig new introductions will be highly challenging for country due to the significant proportion of BH population living abroad.

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.

Biofilms have become a major issue in different spheres of medicine and industry. Source of the issue revolves around increasing resistance of microorganisms towards conventional antibiotics which is even more elevated within biofilms that are difficult to eradicate by means of biocides and antibiotics. Multiple approaches were applied to deal with this issue amongst which green nanotechnology gave promising results and the search for molecules that “freeze bacteria” in the planktonic state. Since silver was used as an antimicrobial agent since ancient times. Considering this a study which deals with the effect of sub-inhibitory concentrations of silver nitrate and silver nanoparticles on biofilm forming capacity of bacteria would yield valuable information to evaluate the effect these substances have on the phenotypical expression of biofilm formation in bacteria as potential biofilm gene expression

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.

Aim To develop an online biofilm calculation tool (Biofilm Classifier), which calculates the optical density cut off value and accordingly determines the biofilm forming categories for the tested isolates by standardized formulas, as well as to compare the results obtained by Biofilm Classifier to manual calculations and the use of predefined values. Methods The biofilm forming capacity of tested strains was evaluated using tissue culture plate method in 96 well plates, and optical density (OD) value of the formed biofilm was measured on an ELISA Microplate reader at 595 nm on a total of 551 bacterial isolates from clinical specimen. Results Comparative analysis indicated that the manual calculation was 100% in accordance with results obtained by the designed software as opposed to the results obtained by use of predefined values for biofilm categorization. When using predefined values compared to manual biofilm categorization for the determination of biofilm positive and biofilm negative strains the specificity was 100%, sensitivity 97.81%, positive predictive value 100%, negative predictive value 96.04% and accuracy 98.57%. Conclusion Considering obtained results, the use of the designed online calculator would simplify the interpretation of biofilm forming capacity of bacteria using tissue culture plate method.

Aim To develop an online biofilm calculation tool (Biofilm Classifier), which calculates the optical density cut off value and accordingly determines the biofilm forming categories for the tested isolates by standardized formulas, as well as to compare the results obtained by Biofilm Classifier to manual calculations and the use of predefined values. Methods The biofilm forming capacity of tested strains was evaluated using tissue culture plate method in 96 well plates, and optical density (OD) value of the formed biofilm was measured on an ELISA Microplate reader at 595 nm on a total of 551 bacterial isolates from clinical specimen. Results Comparative analysis indicated that the manual calculation was 100% in accordance with results obtained by the designed software as opposed to the results obtained by use of predefined values for biofilm categorization. When using predefined values compared to manual biofilm categorization for the determination of biofilm positive and biofilm negative strains the specificity was 100%, sensitivity 97.81%, positive predictive value 100%, negative predictive value 96.04% and accuracy 98.57%. Conclusion Considering obtained results, the use of the designed online calculator would simplify the interpretation of biofilm forming capacity of bacteria using tissue culture plate method.

Spices, which are plant substances used to enhance flavor are at the same time, the most commonly used natural antimicrobial agents in food. Besides this they have shown to effect the biofilm forming capacity of bacteria at different concentrations. In our study we tested the antibacterial effect of different w/V solutions of commercially available spices: cinnamon, curcuma and ginger and investigated their effect on biofilm formation of Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922 and Enterococcus faecalis ATCC 19433. The results of our study indicate that cinnamon had an antibacterial effect on gram positive bacteria, ginger only on E.coli and curcuma did not exhibit any antibacterial properties. Results of the effect of different w/V solutions of spices on biofilm formation of the tested bacteria indicate that the spices had different effects on the tested bacteria and that the applied spice w/V solution did modify biofilm formation of bacteria. Hereby it is evident that the finding of novel antimicrobial compounds should be accompanied by biofilm formation studies since biofilms represent the natural state of bacteria and as such must be taken into consideration. Keywords— Spices, Antimicrobial Effect, Antibiofilm Effect