West Nile virus 'circulation' in Vojvodina, Serbia: Mosquito, bird, horse and human surveillance.

Efforts to detect West Nile virus (WNV) in the Vojvodina province, northern Serbia, commenced with human and mosquito surveillance in 2005, followed by horse (2009) and wild bird (2012) surveillance. The knowledge obtained regarding WNV circulation, combined with the need for timely detection of virus activity and risk assessment resulted in the implementation of a national surveillance programme integrating mosquito, horse and bird surveillance in 2014. From 2013, the system showed highly satisfactory results in terms of area specificity (the capacity to indicate the spatial distribution of the risk for human cases of West Nile neuroinvasive disease - WNND) and sensitivity to detect virus circulation even at the enzootic level. A small number (n = 50) of Culex pipiens (pipiens and molestus biotypes, and their hybrids) females analysed per trap/night, combined with a high number of specimens in the sample, provided variable results in the early detection capacity at different administrative levels (NUTS2 versus NUTS3). The clustering of infected mosquitoes, horses, birds and human cases of WNND in 2014-2015 was highly significant, following the south-west to north-east direction in Vojvodina (NUTS2 administrative level). Human WNND cases grouped closest with infected mosquitoes in 2014, and with wild birds/mosquitoes in 2015. In 2014, sentinel horses showed better spatial correspondence with human WNND cases than sentinel chickens. Strong correlations were observed between the vector index values and the incidence of human WNND cases recorded at the NUTS2 and NUTS3 levels. From 2010, West Nile virus was detected in mosquitoes sampled at 43 different trap stations across Vojvodina. At 14 stations (32.56%), WNV was detected in two different (consecutive or alternate) years, at 2 stations in 3 different years, and in 1 station during 5 different years. Based on these results, integrated surveillance will be progressively improved to allow evidence-based adoption of preventive public health and mosquito control measures.

Testing for viral material in water of public bathing areas of the Danube during summer, Vojvodina, Serbia, 2014.

From August to September 2014 a water quality study was conducted on five popular public Danube beaches in Vojvodina, Serbia. To assess the safety of Danube water for bathing, physical, chemical, bacteriological tests were performed. While many parameters for monitoring the quality of water are regulated by law, there are neither national nor international legislations addressing the presence of viruses in recreational waters. In this study, we performed analyses that surpassed national requirements, and investigated if adenovirus, enterovirus or rotavirus genetic material was present in samples of recreational water collected for quality monitoring. Of 90 water samples obtained during the study, enterovirus material was not found in any sample, but adenovirus and rotavirus genetic materials were respectively detected in 60 and 31 samples. Statistical analyses showed a significant correlation between adenovirus DNA and total coliforms in the water. Even when water samples were adequate for recreational use, adenoviruses were detected in 75% (57/76) of such samples. Our results indicate that implementation of viral indicators in recreational water might be helpful to better assess public health safety. This might be particularly relevant in areas where urban wastewater treatment is insufficient and surface waters affected by wastewater are used for recreation.

Antibiotic Resistance and Probiotics: Knowledge Gaps, Market Overview and Preliminary Screening.

Probiotics are among those products, the use of which is increasing, and they are available primarily as food/dietary supplements, as well as in the form of medicines. This study aims to assess the attitudes and practices of health professionals and students of health sciences, give a short overview of the probiotics currently on the market, and conduct a screening of five food supplements and one drug with respect to antibiotic resistance. Nearly half of the respondents in our survey state that probiotics have no side effects, while only 6.3% believe that the use of probiotics can lead to antibiotic resistance. In addition, more than 40% of the participants throw unused probiotics into municipal waste. The market analysis results indicate that probiotic products on the Serbian market have highly variable CFU counts, while the declared health claims cover numerous beneficial health effects, and they are sometimes even registered as medicines. Lactobacilli are frequently present in probiotic supplements, and are sold in pharmacies and online. The experimental results showed that antibiotic resistance is present in different types of lactobacilli in probiotic products. The risk of using probiotics, regardless of their beneficial health effects, should be taken into account in the future. An update to the regulations governing probiotics, including a stipulation for antimicrobial resistance (AMR) testing, should be established, and guidelines for their proper use and disposal put into place.

Purple Urine Bag Syndrome in a Home-Dwelling Elderly Female with Lumbar Compression Fracture: A Case Report.

Purple urine bag syndrome (PUBS) is an uncommon, but usually benign, underrecognized clinical condition with the distressing presentation of purple, blue or reddish discoloration of a patient's catheter bag and tubing in the setting of catheter-associated urinary tract infections (UTIs). PUBS is the result of the complex metabolic pathway of the dietary essential amino acid tryptophan. Its urinary metabolite, indoxyl sulfate, is converted into red and blue byproducts (indirubin and indigo) in the presence of the bacterial enzymes indoxyl sulfatase and phosphatase. The typical predisposing factors are numerous and include the following: female gender, advanced age, long-term catheterization and immobilization, constipation, institutionalization, dementia, increased dietary intake of tryptophan, chronic kidney disease, alkaline urine, and spinal cord injury (SCI). Here, we present a case of PUBS in a home-dwelling elderly female patient with a history of long-term immobility after a pathological spinal fracture, long-term catheterization, constipation, and malignant disease in remission. Urine culture was positive for Proteus mirabilis. This state can be alarming to both patients and physicians, even if the patient is asymptomatic. Healthcare professionals and caregivers need to be aware of this unusual syndrome as an indicator of bacteriuria in order to initiate proper diagnostics and treatment.

The Effects of Resveratrol-Rich Extracts of Vitis vinifera Pruning Waste on HeLa, MCF-7 and MRC-5 Cells: Apoptosis, Autophagia and Necrosis Interplay.

Resveratrol is a well-studied plant-derived molecule in cancer biology, with a plethora of documented in vitro effects. However, its low bioavailability and toxicity risk hamper its wider use. In this study, vine shoots after pruning were used as a source of resveratrol (RSV). The activity of subcritical water extract (SWE) and dry extract (DE) is examined on three cell lines: HeLa, MCF-7 and MRC-5. The cytotoxic effect is assessed by the MTT test and EB/AO staining, levels of apoptosis are determined by Annexin V assay, autophagia by ULK-1 expression using Western blot and NF-kB activation by p65 ELISA. Our results show that both resveratrol-rich extracts (DE, SWE) have a preferential cytotoxic effect on malignant cell lines (HeLa, MCF-7), and low cytotoxicity on non-malignant cells in culture (MRC-5). Further experiments indicate that the investigated malignant cells undergo different cell death pathways. MCF-7 cells died preferentially by apoptosis, while the HeLa cells died most likely by necrosis (possibly ferroptosis). Protective autophagia is diminished upon treatment with DE in both HeLa and MCF-7 cells, while SWE does not influence the level of autophagia. The extracts are effective even at low concentrations (below IC50) in the activation of NF-kB (p65 translocation).

Medicine Shortages in Serbia: Pharmacists' Standpoint and Potential Solutions for a Non-EU Country.

Medicine shortages in Serbia have evidently been present for several decades, but literature data are scarce. The aim of our study was to get an insight on the present situation in Serbia, review the EU actions when managing shortages, and discern a set of potential measures. A short survey was conducted among 500 pharmacists in public pharmacies, in 23 cities in Serbia. The survey questions addressed frequency of drug shortages, professional actions in the event of shortages, main consequences to patients and pharmacies, putative causes, and pivotal measures for the prevention/mitigation of drug shortages under current conditions. Moreover, a Panel of Experts was organized, whose suggestions and opinions were used to analyze the present situation and to form a set of potential solutions and effective measures to mitigate shortages of medicines. In-depth analysis of current Serbian legislation was conducted, with emphasis on specific steps to be made within the actual legal framework. Examples of good practice in the EU, applicable to a country such as Serbia, were examined. Our research showed that although Serbia is, in some aspects, behind EU countries regarding the approaches to overcome medicine shortages, progress can be made within short period of time, by specific well-targeted actions. Both patients and pharmacists would benefit from it.

West Nile Virus Lineage 2 Associated with Human Case in Republic of Serbia.

A suspicion on West Nile virus (WNV) in Serbia was first reported in 1972 by a seroprevalence study, after which no data were available for four decades. We report full sequence of the isolate obtained for the first time from a human sample in Serbia. The closest clustering was obtained with lineage 2 WNV identified in Greece in 2010. Since WNV lineage 2 emerged in Europe in 2004, a cocirculation of lineages 1 and 2-as observed in Hungary and Italy-cannot be excluded. The reinforcement of surveillance will be required to investigate the possible cocirculation of the two lineages and the burden of WNV in the local population.

Human papillomavirus infection prevalence in female university students in Novi Sad, Serbia.

Introduction: Cervical cancer, attributed to human papillomavirus (HPV) infection, represents the fourth most common and lethal cancer in Serbian women, and the second most common cancer in women aged 15­44. Objective: The aim of the study was to determine the presence of high-risk and low-risk HPV types in population of unvaccinated female university students in Novi Sad, Serbia, and to evaluate possible risk factors for HPV infection. Methods: Sample consisted of 250 young women (19­26 years of age) attending outpatient clinics for screening gynecological examination. All participants in the study completed a specially designed anonymous questionnaire. For the detection of HPV DNA, two commercial kits ­ High Risk HPV Real-TM and Low Risk HPV 6/11 Real-TM (Sacace Biotechnologies, Como, Italy) were used. Thirty positive samples were retested by GenoFlow HPV Array Test (DiagCor Bioscience Incorporation Limited, Hong Kong, China). Results: The overall prevalence rate of HPV was 61.6%. The most common HPV types in the present study were as follows: HPV 16, 31, 51, 52, and 18. Female students with only one sexual partner had significantly lower chance of having HPV infection. Other variables describing lifestyle did not show statistical significance. Conclusion: The present paper provides data on the prevalence of high- and low-risk HPV genotypes among university students in Novi Sad. Obtained results indicate the need for educational activities on sexually transmitted infections, including HPV, together with promotion of healthy lifestyles. According to our results, bivalent and quadrivalent prophylactic vaccines have the potential to prevent over 50% of infections. Percentage of protection with a second-generation prophylactic nonavalent vaccine would be more than 80%.

Frequency of twelve carcinogenic human papilloma virus types among women from the South Backa region, Vojvodina, Serbia.

BACKGROUND/AIM: The aim of this study was to determine the presence and age distribution of different oncogenic human papilloma virus (HPV) types in women in the South Backa region and its relationship to Pap results. MATERIALS AND METHODS: In a group of 1087 women with normal and abnormal cytology, the commercial HR HPV Real-TM kit (Sacace Biotechnologies, Italy) was used. RESULTS: Overall, 50.5% of the women were HPV positive. The presence of HPV types 18, 31, 51, and 58 was significantly influenced by age, while the presence of HPV types 16 and 45 was significantly influenced by cervical cytology. Results of the LSD test show a wide spectrum of high risk HPV among women with normal cytology and women with a low grade cervical lesion rate (atypical squamous cell of undetermined significance (ASCUS) and low grade squamous intraepithelial lesions (LSIL). The most prevalent HPV types found were 16, 31, 51, 18, and 52. In the HSIL group the most prevalent HPV types were 16 and 45. CONCLUSION: The reported results provide new data on the circulation of oncogenic HPV genotypes and frequency of multiple infections among women in Vojvodina and suggest that a prophylactic vaccine against HPV 16 and 18 has the potential to prevent approximately half of the high-grade lesions.

Risk factors for severe influenza A virus infections in post-2009 pandemic period.

Introduction: Literature data concerning risk factors for severe influenza in post-2009 pandemic period, from low- and middle-income Central and Eastern European countries are very limited. Objective: The aim of this study was to investigate the risk factors for severe A(H1N1)pdm09 and A(H3N2) influenza during the post-2009 pandemic period. Methods: During four consecutive seasons of 2010/2011­2013/2014, nasopharyngeal or nasal and pharyngeal swab samples from 153 patients with mild and 147 patients with severe influenza were tested using real-time reverse transcription polymerase chain reaction (real-time RT PCR) assays. Results: The study indicated three statistically significant risk factors of influenza severity, including presence of chronic underlying illness/condition [odds ratio (OR) of 15.2, 95% confidence interval (CI) of 1.8­125.4, p = 0.001), age ≥15 years (OR 9.2, 95% CI 3.5­24.1, p < 0.001), and delay in medical care of more than two days after the symptoms onset (OR 3.2, 95% CI 1.6­6.4, p = 0.001). Conclusion: Obtained results confirmed that patients with chronic underlying illness/condition and older than 15 years had the highest risk for serious complications from influenza and highlighted the importance of start of antiviral therapy within the first two days of illness in order to reduce the risk for the most severe outcomes of influenza, such as acute respiratory distress syndrome and lethal outcome.