Leptospirosis and characterization of Leptospira isolates from patients in Koprivnica-Krizevci County, Croatia from 2000-2004.

Introduction: Leptospirosis, caused by spirochetes of the genus Leptospira , is present in the Koprivnica-Krizevci County area, Croatia. Clinical manifestation can range from asymptomatic, short-term mild, non-specific febrile disease, to severe forms with high mortality rates. Aim: The aim of the study was to valuate culture in front of microscopic agglutination test (MAT) for diagnosis of infection, and to evaluate clinical and laboratory features of the disease. Moreover, we want to characterize Leptospira strains involved in infection in Koprivnica-Krizevci County, Croatia. Methods: We included 68 patients with clinical presentation consistent with leptospirosis collected in a 5-year period (2000-2004). Clinical samples (blood, urine and cerebrospinal fluid, CSF) were inoculated in Kolthoff's medium; species of isolated Leptospira strains was determined with Tm of real-time PCR, serogroup/serovar with MAT and NotI-RFLP analysis. Demonstration of specific antibodies in patients' sera was done using microscopic agglutination test. Results: Leptospira was isolated from the blood of 14/51(27.5 %) patients and the most often identified serogroup/serovar was Icterohaemorrhagiae (8/10, 80%) followed by Grippotyphosa (10%). Regarding to species level, 8/10 isolated belonged to L. interrogans sensu stricto and one to L. kirschneri species. MAT was carried out on 51 patients with suspected leptospirosis, and was positive in 11/51(21.5 %) patients. Most of our patients presented with moderate severe symptoms, were hospitalized from August to October, and were infected mainly during the work or recreation in our county. The frequency of particular clinical features and pathological laboratory findings correlated with the severity of the clinical condition. Conclusions: Leptospirosis can be confirmed microbiologically, culture and MAT contributed almost equally to the diagnosis of infection. Serovar Icterohaemorrhagiae was found as the dominant one, and L. interrogans sensu stricto as dominant species in our county. Epidemiological data shown that leptospirosis occurs seasonally, affects the rural population, and most commonly is presented with moderate severe clinical course.

Haematospirillum jordaniae Cellulitis and Bacteremia.

We isolated Haematospirillum jordaniae from a positive blood culture from a 57-year-old man in Slovenia who had bacteremia and bullous cellulitis of lower extremities. The infection was successfully treated with ciprofloxacin. Our findings signal the need for increased awareness about the clinical course of H. jordaniae and its potential effects as a human pathogen.

Ecology and prevalence of Borrelia burgdorferi s.l. in Ixodes ricinus (Acari: Ixodidae) ticks.

Weather conditions greatly affect tick population densities and activity, on which depends the occurrence of tick-borne diseases (TBDs). During the spring months from 2017 to 2019, 1,357 specimens of Ixodes ricinus ticks were collected at 9 localities in the vicinity of Novi Sad (Serbia). The number of collected ticks varied considerably among the different sampling sites and years. Also, a statistically significant difference was found between months and observed number of ticks for each stadium. By statistical analysis of tick activity depending on microclimatic conditions, a positive and statistically significant relationship between temperature and the number of ticks for each life stage was established, but not for humidity. Dew had a statistically significant impact only on nymphs but not on adults. The infection rate of Borrelia burgdorferi s.l. was the highest in March (46.5-51.2%) and the lowest in May (32.9-34.8%). The highest prevalence was detected in males and the lowest in nymphs. Since there is a positive and statistically significant correlation between tick number and prevalence, the ability to provide weather-based predictions of the seasonal patterns of current tick activity is important for the risk assessment of TBDs such as Lyme borreliosis.

Legionella pneumophila-Epidemiology and Characterization of Clinical Isolates, Slovenia, 2006-2020.

Legionella pneumophila is the causative agent of severe Legionnaires' disease (LD). Although an increasing number of LD cases have been observed, published data from Slovenia are very limited and data on molecular epidemiology are even scarcer. The present retrospective study (2006-2020) reports the results of the microbiological diagnosis of LD, as well as the epidemiology and characterization of the Legionella clinical isolates. We tested urine samples from 15,540 patients with pneumonia symptoms for L. pneumophila infection by urine antigen test, of which 717 (4.6%) tested positive. Isolation of L. pneumophila was successfully performed from 88 clinical specimens, with 82 (93.2%) being identified as L. pneumophila sg 1 and six (6.8%) as L. pneumophila sg 2-14. Sequence-based typing (SBT) identified 33 different sequence types (STs), the most frequent being ST1 and ST23. Sequence type 1 mainly comprised isolates belonging to the Philadelphia subgroup, and ST23 mostly to Allentown/France. The standard SBT scheme, as well as Dresden phenotyping for L. pneumophila, presented a high diversity among isolates.

Evaluation of resazurin-based rapid test to detect colistin resistance in Acinetobacter baumannii isolates.

Acinetobacter baumannii primarily causes colonization, yet it can be an opportunistic pathogen associated with hospital-acquired infections. Many countries report rapid spread of carbapenem-resistant Acinetobacter baumannii (CRAb) which limits treatment options, with colistin frequently being the last line treatment option. The aim of our study was to evaluate a recently developed rapid method, namely the Rapid ResaPolymyxin test, for detection of colistin resistance (ColR) in Acinetobacter baumannii. This test was used for rapid screening of colistin resistance in a clinical setting where there is endemicity of CRAb isolates. A total of 82 A. baumannii clinical isolates were included in the evaluation. The majority of them were resistant to carbapenems (75/82, 91.5%). A total of 37 isolates (45.1%) were resistant to colistin, all being resistant to carbapenems. None of the ColR isolates carried the plasmid-mediated mcr-1 to -5 genes. The Rapid ResaPolymyxin NP test reached a 95.1% categorical agreement with results of reference broth microdilution method, with 93.3% sensitivity and specificity, and positive and negative predictive values being respectively at 92.3% and 97.7%. The Rapid ResaPolymyxin NP test performed well on our collection of clinical and surveillance CRAb isolates from the Central Slovenia region. The test is inexpensive and easy to integrate into laboratory workflow. The main value of the test is rapid categorization of susceptibility and resistance which has important implications with respect to the treatment strategy as well as the infection control measures.