Epidemiology and Clinical Course of Haemorrhagic Fever with Renal Syndrome in New Endemic Area for Hantavirus Infection in Croatia.

BACKGROUND: Hantaviruses remain an important case of emerging and re-emerging infections in human medicine. This study aimed to analyse the epidemiology, clinical presentation, and outcome of hantavirus infections in the western part of Republic of Croatia, a new geographical area for hantavirus infections. METHODS: Retrospective analysis of medical records of patients treated for hemorrhagic fever with renal syndrome (HFRS) at the infectious diseases Clinic of the Clinical Hospital Center in Rijeka, Croatia, from 1 January 2014, to 31 December 2021. RESULTS: During the eight-year period, 251 patients were hospitalized and treated for HFRS, with epidemic outbreaks in years 2014 and 2021. Most patients had a typical clinical course of HFRS and received supportive care. Serological analysis revealed the Puumala Virus (PUUV) as the predominant etiology of the disease. Epidemiological analysis revealed clustering of infections in the region of Gorski Kotar and spread to the area on the Mediterranean coast (Adriatic Sea), which was previously considered an area free from hantavirus infections. CONCLUSIONS: The presented results indicate the spread of hantavirus infections in Croatia from the central low-lying parts of the country to the tourist-attractive western area adjacent to the Mediterranean coast, which was previously considered free of hantavirus infections.

An outbreak of ertapenem-resistant, carbapenemase-negative and porin-deficient ESBL-producing Klebsiella pneumoniae complex.

INTRODUCTION: Carbapenem-resistant Klebsiella pneumoniae is an emerging healthcare-associated pathogen with dynamic molecular epidemiology. This study presents a retrospective analysis of the distribution and antibiotic resistance patterns of ertapenem-resistant ESBL-producing K. pneumoniae strains recovered during an outbreak from 2012 to 2014 in a Croatian University hospital. METHODS: We aimed to estimate genetic relatedness of clinical isolates and underlying mechanisms that conferred the ertapenem-resistant phenotype. RESULTS: Expression analysis of genes involved in the antibiotic resistance showed reduced expression of major non-selective porin channel OmpK35. Reduced expression of OmpK36 porin channel in isolates resistant to at least one more carbapenem, apart from the ertapenem, was found to a lesser degree. Pulsed-field gel electrophoresis analysis of genomic DNA revealed that almost all isolates belonged to the same genetic clone. CONCLUSIONS: Caution regarding ertapenem-resistant, carbapenemase-negative porin-deficient mutants of K. pneumoniae is required as they are widespread, and under selective pressure this could result in a local clonal outbreak.

Clinical, Virological, and Immunological Findings in Patients with Toscana Neuroinvasive Disease in Croatia: Report of Three Cases.

Toscana virus (TOSV) is an arthropod-borne virus, transmitted to humans by phlebotomine sandflies. Although the majority of infections are asymptomatic, neuroinvasive disease may occur. We report three cases of neuroinvasive TOSV infection detected in Croatia. Two patients aged 21 and 54 years presented with meningitis, while a 22-year old patient presented with meningoencephalitis and right-sided brachial plexitis. Cerebrospinal fluid (CSF), serum, and urine samples were collected and tested for neuroinvasive arboviruses: tick-borne encephalitis, West Nile, Usutu, TOSV, Tahyna, and Bhanja virus. In addition, CSF and serum samples were tested for the anti-viral cytokine response. High titers of TOSV IgM (1000-3200) and IgG (3200-10,000) antibodies in serum samples confirmed TOSV infection. Antibodies to other phleboviruses (sandfly fever Sicilian/Naples/Cyprus virus) were negative. CSF samples showed high concentrations of interleukin 6 (IL-6; range 162.32-2683.90 pg/mL), interferon gamma (IFN-γ; range 110.12-1568.07 pg/mL), and IL-10 (range 28.08-858.91 pg/mL), while significantly lower cytokine production was observed in serum. Two patients recovered fully. The patient with a brachial plexitis improved significantly at discharge. The presented cases highlight the need of increasing awareness of a TOSV as a possible cause of aseptic meningitis/meningoencephalitis during summer months. Association of TOSV and brachial plexitis with long-term sequelae detected in one patient indicates the possibility of more severe disease, even in young patients.

Tick-borne encephalitis outbreak following raw goat milk consumption in a new micro-location, Croatia, June 2019.

In June 2019, the Croatian Institute of Public Health was informed of a cluster of patients with laboratory confirmed tick-borne encephalitis (TBE) from the Gorski Kotar region. Five of the six patients with TBE reported consuming raw (unpasteurized) goat milk in the two week period before symptom onset, and one reported a recent tick bite. To assess risk factors for infection, we selected six control individuals from among healthy family and community members, and conducted a case-control analysis. None of the cases or controls were vaccinated against TBE. Individuals with TBE (cases) had 25 (95 % CI 0.8-1410.2, p = 0.021) times higher odds of raw goat milk consumption compared to healthy controls. Milk samples from 12 goats from the implicated farm were tested for the TBE virus (TBEV) using RT-PCR. TBEV RNA was not detected in the milk, but serological testing of goats and other farm animals yielded evidence of exposure to the virus: Six goats from the flock had TBEV neutralizing antibodies. Our findings suggest that the vehicle for the outbreak was raw goat milk from a single farm. Following public health advice to cease consumption of raw dairy products, no further cases have been reported.

Challenges to antimicrobial susceptibility testing of plant-derived polyphenolic compounds.

As multidrug resistance gains momentum, the last two decades have seen an ever-growing interest in the antimicrobial properties of plant extracts and plant-derived compounds. Most of the focus is on polyphenols - a large and diverse group of phytochemicals with strong antibacterial activity. Testing methods provide reliable results as long as they follow standard procedures. However, methods and procedures used in antimicrobial susceptibility testing (AST) are often too diverse to allow comparison of results. The lack of uniformity and comparability is much owed to the absence of guidelines. The focus of this review is to give a critical overview of different methods used in the assessment of polyphenols antimicrobial efficacy and to highlight the importance of their standardisation.