•Resistance is prevalent among slow growing nontuberculous mycobacteria from animals.•Nearly all isolates showed multidrug resistance.•Humans are at risk of zoonosis that may be difficult to treat ...with current therapy.
Non-tuberculous mycobacteria are opportunistic pathogens that cause disease mainly in immunocompromised hosts. The present study assessed the prevalence of antibiotic resistance among such mycobacteria from domestic and wild animals in Croatia sampled during several years within a national surveillance program.
A total of 44 isolates belonging to nine slow-growing species were genotyped and analyzed for susceptibility to 13 antimicrobials often used to treat non-tuberculous mycobacterial infections in humans.
Most prevalent resistance was to moxifloxacin (77.3%), doxycycline (76.9%), and rifampicin (76.9%), followed by ciprofloxacin (65.4%), trimethoprim-sulfamethoxazole (65.4%), and linezolid (61.4%). Few isolates were resistant to rifabutin (7.7%) or amikacin (6.8%). None of the isolates was resistant to clarithromycin. Nearly all isolates (86.4%) were resistant to multiple antibiotics.
Our findings suggest substantial risk that human populations may experience zoonotic infections with non-tuberculous mycobacteria that will be difficult to treat using the current generation of antibiotics. Future work should clarify how resistance emerges in wild populations of non-tuberculous mycobacteria.
Non-tuberculous mycobacteria (NTM) are opportunistic pathogens capable of causing infections in humans and animals. The aim of this study was to demonstrate the potential role of domestic and wild ...animals as a reservoir of multiple resistant, rapidly growing NTM strains representing a potential zoonotic threat to humans. A total of 87 animal isolates belonging to 11 rapidly growing species (visible colonies appear within three to seven days) were genotyped and tested for susceptibility to the 15 most commonly used antibiotics in the treatment of such infections in a human clinic. By determining the antimicrobial susceptibility, the most prevalent resistance was found to cephalosporins (>50%), followed by amoxicillin–clavulanate (31.0%), clarithromycin (23.0%), tobramycin (14.9%) and doxycycline (10.3%). Resistance to imipenem, ciprofloxacin, minocycline and linezolid was notably lower (<7.0%). All tested isolates were susceptible to amikacin and moxifloxacin. The most frequent resistance was proved in the most pathogenic species: M. fortuitum, M. neoaurum, M. vaccae and M. porcinum. Meanwhile, other species displayed a higher sensitivity rate. No significant resistance differences between domestic and wild animals were found. The established significant frequency of resistance highlights the significant zoonotic potential posed by circulating rapidly growing NTM strains, which could lead to challenges in the treatment of these infections.
To investigate the degree of genetic variability of bovine leukemia virus (BLV) strains circulating in Croatia, 29 isolates from the six largest dairy farms were examined by PCR for a segment of the ...gp51
env
gene, followed by DNA sequencing and phylogenetic analysis. The nucleotide sequences were compared with other previously characterized BLV strains from different geographical areas, comprising all seven known BLV genotypes. The Croatian sequences showed six to eight nucleotide substitutions: six silent substitutions and two amino acid changes. Four of those substitutions were within epitopes. In comparison to the sequences of other BLV genotypes, our isolates showed the closest relationship to genotype 1 isolates PL-3252 (FJ808585) and AL-148 (FJ808573) from Argentina. The degree of variation between our sequences and those of genotype 1 was 0.2- 4.6 %. In phylogenetic trees based on 400-nt and 519-nt sequences, all of the Croatian sequences clustered separately from the other sequences, revealing a new genotype.
Canine respiratory coronavirus is a relatively new addition to the list of pathogens causing canine infectious disease complex. The virus is highly contagious, with a high prevalence in the dog ...population worldwide, especially in shelters. This study aimed to establish the presence and risk factors associated with infection in privately owned dogs and breeding colonies. This study was the first to demonstrate the presence of canine respiratory coronavirus in Croatia. Out of the 257 serum samples, 35.03% of dogs from breeding kennels and 43% of pet dogs tested enzyme-linked immunoassay positive, but the difference was not statistically significant. Sex was not an important risk factor, but the seropositivity rate increased with age. Mixing of dogs during hunting, training and dog shows was not associated with a higher seroprevalence in the breeding colonies. Daily cleaning and disinfection showed little effect on the infection spread. The study was done on a limited sample. However, it still provides evidence that the epizootiology of this disease is complex. There is no available vaccine for canine respiratory coronavirus, and further studies on environmental and risk factors will give the valuable data needed to prevent this disease. Keywords: canine respiratory coronavirus; seroprevalence; breeding colonies; ELISA; Croatia Pseci respiratorni koronavirus nedavno je dodan na listu patogena koji uzrokuju zarazni kašalj legla. Zbog svoje izrazite kontagioznosti proširio se diljem svijeta, a osobito je cest u pasa u skloništima za nezbrinute životinje. Osnovni je cilj ovog istraživanja bio ispitati proširenost infekcije u pasa u privatnom vlasništvu i uzgajivacnicama te ustanoviti cimbenike rizika koji pogoduju infekciji. Ovo je bilo prvo istraživanje koje je dokazalo prisutnost pseceg respiratornog koronavirusa u Hrvatskoj. Od 257 pretraženih uzoraka seruma 35,03% uzoraka pasa privatnih vlasnika i 43% uzoraka pasa iz uzgajivacnica dalo je pozitivan rezultat imunoenzimnog testa. Važno je napomenuti da razlika u seroprevalenciji izmedu ovih dviju populacije nije bila statisticki znakovita. Spol takoder nije bio znakovit cimbenik rizika, ali je seroprevalencija rasla s porastom dobi životinja. Miješanje životinja iz razlicitih uzgoja, prilikom lova, dresure ili izložbi pasa, nije dovelo do porasta seroprevalencije u uzgajivacnicama. Ni provodenje dnevne dezinfekcije nije znakovito utjecalo na seroprevalenciju. Ovo je istraživanje provedeno na ogranicenom broju uzoraka, ali ipak dokazuje složenost epizootiologije infekcije psecim respiratornim koronavirusom. Kako cjepivo još uvijek nije dostupno, dodatna istraživanja cimbenika okoliša i domacina, važnih za epizootiologiju ove bolesti, dat ce važne podatke nužne za provodenje mjera prevencije. Kljucne rijeci: pseci respiratorni koronavirus; seroprevalencija; uzgajivacnice; ELISA; Hrvatska
West Nile Virus Neuroinvasive Disease (WNV NID) requires prolonged intensive care treatment, resulting in high mortality and early disability. Long-term results are lacking. We have conducted an ...observational retrospective study with a prospective follow-up of WNV NID patients treated at the Intensive Care Unit (ICU), University Hospital for Infectious Diseases, Zagreb, Croatia, 2013-2018. Short-term outcomes were vital status, length of stay (LOS), modified Rankin Scale (mRS), and disposition at discharge. Long-term outcomes were vital status and mRS at follow-up. Twenty-three patients were identified, 78.3% males, median age 72 (range 33-84) years. Two patients (8.7%) died in the ICU, with no lethal outcomes after ICU discharge. The median ICU LOS was 19 days (range 5-73), and the median hospital LOS was 34 days (range 7-97). At discharge, 15 (65.2%) patients had moderate to severe/mRS 3-5, 6 (26.0%) had slight disability/mRS 2-1, no patients were symptom-free/mRS 0. Ten (47.6%) survivors were discharged to rehabilitation facilities. The median time to follow-up was nine months (range 6-69). At follow-up, seven patients died (30.5%), five (21.7%) had moderate to severe/mRS 3-5, one (4.3%) had slight disability/mRS 2-1, six (26.1%) had no symptoms/mRS 0, and four (17.4%) were lost to follow-up. Briefly, ten (43.5%) survivors improved their functional status, one (4.3%) was unaltered, and one (4.3%) aggravated. In patients with severe WNV NID, intensive treatment in the acute phase followed by inpatient rehabilitation resulted in significant recovery of functional status after several months.
In a total of 1536 blood serum samples analysed by ELISA, antibodies for IAV nucleoprotein (NP) were detected in 30.3%. Results from HI show that the most common subtype of swIAV in the Croatian pig ...population was H1N1 (44.6%), followed by H3N2 (42.7%) and H1N2 (26.3%). Antibodies to at least one subtype were detected in 62.19% of blood serum samples. Detection of swIAV antigen was performed by IHC and detected in 8 of 28 lung samples collected post-mortem. The matrix (M) gene was detected in nine of one hundred and forty-two lung tissue samples and in seven of twenty-nine nasopharyngeal swabs. Phylogenetic analysis of amplified HA and NA gene fragments in Croatian isolates suggests the presence of swIAV H1avN1av.
The data on the coronavirus disease (COVID-19) in solid-organ transplant recipients (SOTRs) in Croatia is unknown. The aim of this study was to analyze the seroprevalence of severe acute respiratory ...syndrome coronavirus 2 (SARS-CoV-2) in Croatian SOTRs.
From 7 September to 27 November 2020 (beginning of the second COVID-19 pandemic wave), a cross-sectional screening for COVID-19 was performed in the adult outpatient liver (LTRs; N = 280) and kidney transplant recipients (KTRs; N = 232). Serum samples were initially tested for SARS-CoV-2 IgG antibodies using a commercial enzyme-linked immunosorbent assay (ELISA; Vircell Microbiologists, Granada, Spain). All positive samples were confirmed using a virus neutralization test (VNT). Data on risk exposure and COVID-19 related symptoms were collected using a questionnaire.
The transplanted cohort's seroprevalence detected by ELISA and VNT was 20.1% and 3.1%, respectively. Neutralizing (NT) antibodies developed in 15.6% of anti-SARS-CoV-2 ELISA IgG positive SOTRs. The difference in seropositivity rates between LTRs and KTRs was not statistically significant (ELISA 21.1%
. 19.0%, P = 0.554; VNT 3.6%
. 2.6%, P = 0.082). Overall VNT positivity rates were higher in patients who reported participation in large community events (5.9%
. 1.0%; P = 0.027) as well as in patients who reported COVID-19 related symptoms in the past six months. In addition, symptomatic VNT positive patients showed significantly higher (P = 0.031) NT antibody titers (median 128, interquartile range (IQR) = 32-128) compared to asymptomatic patients (median 16, IQR = 16-48).
This study showed that 15.6% of anti-SARS-CoV-2 ELISA positive Croatian SOTRs developed NT antibodies indicating protective immunity. Further studies are needed to determine the dynamic of NT antibodies and COVID-19 immunity duration in immunocompromised populations such as LTRs and KTRs.
West Nile virus (WNV) is an emerging zoonotic pathogen with rapid global expansion. The virus circulation is confirmed in many countries of Mediterranean Basin and Southern and Central Europe. In our ...study detection of specific WNV antibodies was performed in horses and cattle sera samples collected from October 2010 to April 2011. Serum samples were randomly taken from different parts of Croatia and tested by IgG and IgM ELISA. Positive serological results were confirmed by virus neutralization assay (VN-assay) and plaque reduction neutralization test (PRNT). Results showed that WNV antibodies were present in 72 out of 2098 horse sera (3.43%) and 3 of 2695 cattle sera (0.11%). The highest seroprevalence was found in Eastern Croatia in counties next to Hungarian, Serbian and Bosnia and Herzegovinian state borders. In Adriatic part of Croatia positive animals were found only in the westernmost county, near Slovenian and Italian borders. Geographic distribution and number of positive horses indicated that WNV is highly present in Croatia and spreading from East to West. However, positive horses in westernmost part of country indicate possible second origin of spreading. Location of serological positive cattle supports the hypothesis that seropositive cattle could be indicators of high WNV activity in the respective geographic regions.
Hepatitis E (HE) is an emerging public health problem in developed countries worldwide. Nowadays, it is known that the hepatitis E virus (HEV) has zoonotic potential, whereas domestic pigs and wild ...boars are recognized as the true reservoirs of the disease. Therefore, the aim of our study was to define the seroprevalence and viral persistence of HEV in pigs and wild boars in Croatia. In total, 1424 samples originating from domestic pigs from 14 counties, and 1000 samples of wild boar origin, collected from 16 counties, were tested during 2016 for the presence of HEV antibodies using a commercially available enzyme linked immunosorbent assay (ELISA). All positive samples, as well as samples originating from in-contact domestic pigs and wild boars, were further tested for the presence of HEV RNA using a real time RT-PCR protocol, followed by a nested RT-PCR protocol as a confirmatory test. In 11 of 14 counties, 469 seropositive domestic pigs were found with an overall seroprevalence of 32.94% (95% CI 30.55-35.42), ranging from 8.33% to 60.00%, depending on their geographical origin. Seropositive wild boars were found in six out of 16 counties, showing an overall seroprevalence of 31.10% (95%CI 28.31-34.04%), ranging from 7.70% to 50.60%. The highest seroprevalence was found in the Vukovar-Srijem and Osijek-Baranja Counties (North-East Croatia) where pig breeding dominates and the highest density of wild boars is recorded. While no positive HEV RNA samples were detected in tested domestic pigs, seventeen serologically positive wild boars (11.33%; 95%CI 7.19-17.14) younger than one year of age were found to be HEV RNA positive, indicating chronic infection and possible prolonged virus spread into the environment. The detected viral presence despite the immune response indicates that wild boars have a key role in HEV epidemiology. Key words: hepatitis E, domestic pig, wild boar, seroprevalence, viraemia
This study was performed on 29 domestic cats with a variety of clinical signs, possibly related to FIV infection. Blood samples were tested by a rapid immunochromatographic (ICA) procedure for ...detection of FIV antibodies. Subsequently, polymerase chain reaction (PCR) was performed to amplify a portion of the proviral gag gene. All 11 positive PCR products were sequenced and compared with previously reported FIV sequences. Croatian proviral isolates that could be amplified were clustered within subtype B, and additional heterogeneity was confirmed by the formation of three separate clusters. Phylogenetic analysis of circulating strains in Croatia and in southeast Europe is necessary to improve diagnostic methods and selection of the appropriate vaccinal strains.
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