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.

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 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