In Vitro Efficacy of Topical Antimicrobial Drugs

CILJ ISTRAŽIVANJA: Cilj ovoga rada je utvrditi osjetljivost bakterija koje najčešće izazivaju lokalizirane infekcije u KBC Split tijekom 2020. godine na pojedine topikalne antimikrobne lijekove kako bi se ustanovilo koji od testiranih antibiotika je najbolji izbor za empirijsko liječenje ovih infekcija. MATERIJALI I METODE: Retrospektivnim istraživanjem analizirani su podatci o osjetljivosti na topikalne antimikrobne lijekove (aminoglikozide, fluorokinolone, kloramfenikol, kolistin te mupirocin) pojedinih vrsta bakterija koje su najčešći uzročnici lokaliziranih infekcija: Escherichia coli i ESBL E. coli, Klebsiella pneumoniae (uključujući i ESBL sojeve), Pseudomonas aeruginosa, Acinetobacter baumannii te Staphylococcus aureus (uključujući MRSA). Osjetljivost je ispitivana metodom disk difuzije, a za kolistin metodom dilucije u bujonu. REZULTATI: Rezultati pokazuju visoku rezistenciju bakterije Escherichia coli na antibiotike ciprofloksacin i levofloksacin, a nisku na kloramfenikol i kolistin. U usporedbi s Escherichia coli, ESBL producirajući soj Escherichia coli pokazuje znatno visoku rezistenciju na sve ispitivane antibiotike osim na kolistin i kloramfenikol. Sojevi Klebsiella penumoniae su bili visoko rezistentni na antibiotik tobramicin, a nisko rezistentni na kloramfenikol i kolistin. ESBL producirajući sojevi ove bakterije su imali znatno visoku rezistenciju na sve ispitivane antibiotike osim na kloramfenikol i kolistin. Pseudomonas aeruginosa je bio visoko rezistentan na ciprofloksacin i levofloksacin, a nisko na kolistin. Sojevi Acinetobacter baumanni su bili visoko rezistentni na levofloksacin, a nisko na kolistin. Izolati Staphylococcus aureus su bili visoko rezistentni na levofloksacin, a dobro osjetljivi na kloramfenikol i norfloksacin. MRSA sojevi su bili visoko rezistentni na levofloksacin, u značajno većim postotcima od MSSA sojeva, a dobru osjetljivost pokazali su na mupirocin. ZAKLJUČAK: Podatci ove studije pokazuju visoke stope rezistencije ispitivanih bakterija na pojedine topikalne antimikrobne lijekove, što je u skladu s trendovima porasta rezistencije bakterija u svijetu, te možemo pretpostaviti da će se ta rezistencija u budućnosti još povećavati.AIM OF THE RESEARCH: The main aim of this research was to determine the susceptibility of bacterial isolates which are that most frueqent causative agents of localized infections in UHC Split in 2020 to certain topical antimicrobial drugs and to determine which of the antibiotics tested is the best choice for empirical treatment of these infections. MATERIAL AND METHODS: In this retrospective study the data of susceptibility to topical antimicrobial drugs (aminoglycosides, fluoroquinolones, chloramphenicol, colistin and mupirocin) of certain types of bacteria that are the most common causes of localized infections were analyzed. Bacterial isolates included were Escherichia coli and ESBL E. coli, Klebsiella pneumoniae (including Klebsiella pneumoniae ESBL strains), Pseudomonas aeruginosa, Acinetobacter baumannii, and Staphylococcus aureus (including MRSA). Sensitivity was examined by disk diffusion method, and for colistin by broth dilution method. RESULTS: The results showed high resistance rates of Escherichia coli to the antibiotics ciprofloxacin and levofloxacin, and the low rates to chloramphenicol and colistin. Compared with Escherichia coli, the ESBL-producing strain of Escherichia coli showed significantly high resistance to all tested antibiotics except colistin and chloramphenicol. Strains of Klebsiella penumoniae were high resistant to the antibiotic tobramycin, and low resistant to chloramphenicol and colistin. ESBL-producing strains of this bacterium were significantly high resistant to all tested antibiotics except chloramphenicol and colistin. Pseudomonas aeruginosa was high resistant to ciprofloxacin and levofloxacin, and low to colistin. Acinetobacter baumanni strains were high resistant to levofloxacin and low to colistin. Staphylococcus aureus isolates were high resistant to levofloxacin and sensitive to chloramphenicol and norfloxacin. MRSA strains were high resistant to levofloxacin in significantly higher percentages than MSSA strains, and showed the good sensitivity to mupirocin. CONCLUSION: The data from this study follow the global trends of increasing resistance, and can be assumed that this number will increase in the future

Prvi hrvatski pedijatrijski bolesnik s Geotrichum capitatum respiratornom infekcijom

Geotrichum capitatum can cause infections in humans and its importance has recently been recognized in patients with immunosuppressive conditions. In this report we present the first Croatian pediatric patient with pneumonia in whom G. capitatum was isolated in three sequential bronchoalveolar lavage and tracheal aspiration specimens.Geotrichum capitatum može uzrokovati infekcije u ljudi i njegov značaj je nedavno prepoznat u imunosuprimiranih bolesnika. U ovom radu prikazujemo prvog hrvatskog pedijatrijskog bolesnika s pneumonijom u kojeg je G. capitatum izoliran u tri uzastopna bronhoalveolarna lavata i aspirata traheje

Kolonizacija vankomicin rezistentnom bakterijom Enterococcus faecium i Clostridium difficile infekcija u hematološke bolesnice

Vancomycin-resistant enterococci (VRE), especially Enterococcus faecium, have emerged as significant nosocomial pathogens and patients with impaired host defenses are at a particular risk of VRE infection. The most common occurrence is asymptomatic colonization of the gastrointestinal tract that can persist for a long time and serve as a reservoir for transmission of VRE to other patients. We present a case of a patient who was diagnosed with acute myelogenous leukemia and suffered from bone marrow aplasia following induction therapy. The patient received prolonged broad-spectrum antimicrobial therapy. During hospital stay, the patient developed Clostridium difficile infection (CDI) and was found to be colonized with a strain of Enterococcus faecium resistant to vancomycin during therapy for CDI. This case also highlights the role of risk factors that could contribute to development of resistance, particularly CDI. Early detection of VRE colonization or infection is a crucial component in hospital program designed to prevent transmission of nosocomial infections. Surveillance cultures of such patients should be mandatory.Vankomicin-rezistentni enterokoki (VRE), naročito Enterococcus faecium, spadaju među najznačajnije bolničke patogene, pri čemu su naročito ugroženi bolesnici oslabljenog imunosnog statusa. Pritom je najčešća pojava asimptomatske kolonizacije probavnog sustava koja može ustrajati duže vremena i biti rezervoar za širenje VRE na ostale bolesnike. Donosimo prikaz slučaja bolesnice s dijagnozom akutne mijelomonocitne leukemije praćene aplazijom koštane srži nakon indukcijske terapije. Bolesnica je liječena antibioticima širokog spektra. Tijekom hospitalizacije u bolesnice se razvila infekcija bakterijom Clostridium difficile (CDI) uz dokazanu kolonizaciju sojem Enterococcus faecium rezistentnog na vankomicin tijekom terapije zbog CDI. Također su prikazani čimbenici rizika koji su u navedenom slučaju mogli poslužiti kao potencijalni okidač za razvoj rezistencije, s osobitim naglaskom na CDI. Rano otkrivanje kolonizacije ili infekcije navedenim sojevima je iznimno značajan čimbenik bolničkog programa za prevenciju širenja bolničkih infekcija. Mikrobiološki nadzor uzimanjem nadzornih kultura mora biti obvezni dio protokola pri hospitalizaciji takvih bolesnika

Klinički prikaz genitourinarne tuberkuloze sa sumnjom na abdominalnu neoplazmu

Genital tuberculosis is a rare and unexpected disease in European countries including Croatia. Diagnosis of female genital tract tuberculosis is challenging and is rarely pin-pointed by clinical symptoms because of their low specificity. The authors decided to present a case of genitourinary tuberculosis in a young, immunocompetent fertile woman with high clinical suspicion of abdominal tumor mass. Although considered a desease of the past, rare clinical presentation of genital tuberculosis should be expected and taken into account.Genitalna tuberkuloza rijetka je i neočekivana bolest u zemljama Europske unije uključujući i Hrvatsku. Postavljanje dijagnoze tuberkuloze genitalnog sustava otežano je nespecifičnim simptomima. Prikazujemo slučaj genitourinarne tuberkuloze kod mlade imunokompetentne žene generativne dobi gdje je klinička dijagnoza upućivala na abdominalnu neoplazmu. U vremenu velikih društvenih promjena prisutnih posljednjih godina rijetke kliničke prezentacije tuberkuloze i dalje su moguće kao diferencijalna dijagnoza

CHARACTERISTICS OF CHILDREN TUBERCULOSIS IN UNIVERSITY HOSPITAL CENTRE SPLIT FROM 1990 TO 2012

Tuberkuloza u dječjoj dobi posebna je bolest po svojim epidemiološkim i kliničkim osobitostima. Najučinkovitija mjera u suzbijanju širenja bolesti jest brzo otkrivanje te neodgodivo i učinkovito liječenje oboljelih. Kako najveći dio djece ima mikroskopski negativne iskašljaje, brzo postavljanje dijagnoze može biti problem. U ovom radu prikazali smo učestalost, liječenje, dijagnostiku i kliničke osobitosti djece liječene u KBC-u Split tijekom 22 godine.Childhood tuberculosis is a unique disease regarding epidemiological and clinical features. The most effective measures to combat the spread of the disease are rapid detection and prompt and effective treatment. As most of the children have negative sputum microscopy, rapid diagnosis can be challenging. In this paper, the data on incidence, treatment, diagnostic and clinical characteristics of children treated for tuberculosis in the University Hospital Centre Split during 22 years are presented

Comparison of the novel Uroquattro HB&L™ system and classical phenotypic method for rapid screening of multidrug-resistant organism colonization at the University Hospital Centre Split, Croatia

Uvod: Infekcije uzrokovane višestruko otpornim organizmima (MDRO) se teško liječe i često imaju nepovoljan ishod za bolesnika. Budući da infekciji najčešće prethodi kolonizacija, rana identifikacija koloniziranih bolesnika je neophodna u prevenciji nastanka infekcije. Za probir MDRO najčešće se koriste kultivacijske metode kojima vrijeme potrebno za izdavanje nalaza (TAT) iznosi od 48 do 72 h. Cilj ove studije bio je usporediti rezultate nove, brze, poluautomatske metode za detekciju MDRO (HB&L Uroquattro, Alifax) sa standardnom kultivacijom na selektivnim podlogama. Metode/materijali: Tijekom dva nasumično odabrana dana, bolesnicima hospitaliziranima u Kliničkom bolničkom centru Split, uzorkovano je ukupno 21 aksilarnih, 19 rektalnih te 20 briseva vestibuluma nosa. Svaki obrisak je prikupljen u duplikatu, jedan za kultivacijsku, a drugi za brzu poluatomatsku metodu. Brisevi su kultivirani i MDRO izolati identificirani na selektivnim podlogama (Chromagar MRSA i Chromagar ESBL) prema standardnom laboratorijskom protokolu. Nova Alifaxova metoda je provedena prema uputama proizvođača. Rezultati: U usporedbi s klasičnom metodom, TAT nove metode je bio značajno kraći (6,5 h). Klasičnom metodom je na Chromagar ESBL selektivnom mediju izolirano 10 sojeva koji stvaraju ß-laktamaze proširenog spektra (ESBL) iz 10 različitih rektalnih uzoraka. Isti uzorci su bili pozitivni na prisustvo ESBL- producirajućih bakterija brzom Alifaxovom metodom. Kultivacijom na selektivnom Chromagar MRSA mediju i brzom metodom otkriveno je 11 uzoraka pozitivnih na prisustvo MDRO. Zaključak: Visoki stupanj podudarnosti otkrivanja pozitivnih i negativnih uzoraka dobiven klasičnom i novom metodom, kao i značajno skraćivanje TAT-a novom metodom je ohrabrujući. Budući je studija provedena na relativno malom uzorku, potrebno je istraživanje proširiti na veći broj ispitanika.Background. Infections caused by multidrug-resistant organisms (MDRO) are difficult to treat and associated with poor outcomes for patients. Therefore, early identification and management of colonization are essential as first steps in infection prevention. Culture-based methods have been widely used for MDRO screening. The turnaround time (TAT) for the identification of carriers varies between 48-72 h with this method. The aim of our study was to compare the performance of the new rapid semiautomatic method for detection of MDRO (HB&L Uroquattro, Alifax) with standard cultivation on selective media. Methods. Twenty-one axillary, 20 nose and 19 rectal swabs were taken in duplicate on two selected days at the University Hospital Centre Split, Croatia. Swabs were cultivated and MDRO isolates were identified on selective media (Chromagar MRSA and Chromagar ESBL) according to the standard operating procedure. Novel Alifax method was performed according to manufacturer’s instructions. Results. TAT for the new method was significantly lower (6.5 h) in comparison to the classical method. With classical method, 10 extended spectrum ß-lactamases (ESBL) producing strains from 10 different rectal specimens were isolated on Chromagar ESBL media. Exactly the same specimens were positive for the presence of ESBL-producing bacteria by rapid Alifax method. On selective Chromagar MRSA media, 11 MDRO were isolated, while rapid method detected 11 MDRO from the same specimens. Conclusions. High concordance of positive and negative results obtained with classical and rapid method is encouraging. However, our study was performed on a small sample size and further research with larger sample size is needed

Distribution of rotavirus genotypes in three Croatian regions among children ≤5 years of age (2012–2014)

Objectives: Rotavirus is the major cause of severe diarrhea in young children worldwide. In countries like Croatia, where rotavirus vaccine has not been introduced in the national immunization program, prospective surveillance is necessary to establish the diversity of rotavirus strains. The aim of this study was to describe the prevalence and geographical distribution of rotavirus strains in Croatia and to detect the possible emergence of novel strains. ----- Methods: The study was conducted among children ≤5 years of age with acute gastroenteritis at three hospitals located in different geographical regions of Croatia, during the years 2012 to 2014. Rotavirus was detected in stools using an immunochromatographic assay and then sent for further molecular analysis. ----- Results: Genotyping of 822 rotaviruses showed that the predominant circulating strain was G1P[8] (61.9%), followed by G2P[4] (19.5%), G1P[4] (3.9%), and G3P[8] (2.9%). A high prevalence of reassortants among common human rotavirus genotypes was detected (7.7%). Possible zoonotic reassortants were found, including G8 and G6 strains. The latter is described for the first time in Croatia. ----- Conclusions: This study represents pre-vaccination data that are important for decisions regarding immunization strategies in Croatia. The high prevalence of 'common' rotavirus strains circulating in Croatia may advocate for rotavirus vaccine introduction, but further surveillance is necessary to monitor the possible emergence of novel genotypes

Characterization of the extended-spectrum β-lactamases and determination of the virulence factors of uropathogenic Escherichia coli strains isolated from children

BACKGROUND AND AIM: The aim of the study was to characterize ESBL-producing uropathogenic Escherichia coli (UPEC) strains isolated in children. That included the investigation of virulence factors and the analysis of the types of β-lactamases at the molecular genetic level. ----- MATERIAL AND METHODS: During the 2-year study period, 77 ESBL-producing E. coli strains were recovered from urine samples of febrile children with significant bacteriuria hospitalized at one Croatian hospital. Susceptibility of isolates to bactericidal serum activity was tested by Shiller and Hatch method, while adhesin expression was determined by agglutination methods. Characterization of ESBLs was performed by PCR with specific primers for ESBLs and by sequencing of bla (ESBL) genes. Genotyping of the E. coli isolates was performed by pulsed-field gel electrophoresis (PFGE). ----- RESULTS: Twenty-seven (35.1 %) and 50 (64.9 %) ESBL-producing UPEC strains were isolated in neonates and infants, respectively. Of 70 strains investigated for the presence of virulence factors, adhesins were detected in 48.6 % strains (8.6 % in the neonate and 40 % in the infants group) giving a statistically significant difference in adhesin expression between the two groups (p < 0.01). Hemolysin was produced by 84.3 %, whereas 70 % of strains were serum-resistant. The bla (TEM) gene was detected in 22 (28 %) and bla (SHV) gene in 57 strains (74 %), whereas bla (CTX-M) gene was detected in only two isolates (2.5%). In ten isolates, bla (TEM) and bla (SHV) were simultaneously detected. Sequencing of bla (SHV) genes revealed that SHV-5 β-lactamase was by far the most prevalent and was found in 51 strains (66 %). The strains were clonally related as demonstrated by PFGE and assigned into ten clusters. ----- CONCLUSIONS: Infection control measures should be employed and the consumption of expanded-spectrum cephalosporins in the hospital should be restricted

Measles cases in Split-Dalmatia County (a Croatian tourist region), in May–July 2019: outbreak report and lessons learnt

Background: Measles elimination was accomplished in Croatia in 2016. Split-Dalmatia County, with population of ca. 425 000 inhabitants, is among the most important Croatian tourist areas with numerous seasonal workers coming during summer months. In both 2018 and 2019, more than 3 million tourists visited this county. In 2000-2018, there were no measles cases in this county, or their number was low (1-3 cases per year). Methods: After measles was clinically suspected, all contacts were traced and contacted. Detection of specific IgM/IgG antibodies and real-time reverse transcription-polymerase chain reaction detection of viral RNA were used for laboratory confirmation. Sequencing and genotyping were performed for strains' molecular epidemiology analysis. Results: Six epidemiologically unlinked measles virus occurrences happened in Split-Dalmatia County in 15 May-19 July 2019. Causative viral strains belonged to genotypes B3 and D8. Four were single imported cases. Ten patients belonged to two separate clusters within domicile population. Multiple individual and public health measures were implemented. In total, 483 contacts were identified, 64.2% within healthcare system where two persons contracted the disease. Conclusions: Besides the importance of timely vaccination of children, the lessons learned from this outbreak point to the need of stricter implementation of other aspects of Croatian measles prevention programme, such as checking of vaccination status in early adulthood. Despite the fact that measles elimination within domicile population in this tourist region has been accomplished and maintained for years, continuous public health workers' efforts are still necessary for identification and diminishment of populational pockets of susceptibility