A total of 3543 raw cow milk samples were collected in three regions of Croatia: western, eastern and other regions during four seasons. Samples were measured for aflatoxin M1 (AFM1) concentrations ...using the enzyme immunoassay method. Elevated levels (>50 ng/kg) of AFM1 were analysed by validated liquid chromatography with triple quadruple mass spectrometry (LC-MS/MS). The limits of detection (LOD) and quantification (LOQ) of the LC-MS/MS method were 7.3 and 28 ng/kg, respectively. The mean AFM1 levels measured in the three regions over four seasons were in the ranges (ng/kg): eastern Croatia 7.25–26.6; western Croatia 5.91–9.26; other regions of Croatia 7.17–13.6. The highest incidence of samples exceeding the EU MRL (50 ng/kg) of 9.32% was measured in autumn (October–December) in the eastern region. Only eight samples were found to exceed the EU MRL in winter. The highest AFM1 levels were measured in December (764.4 ng/kg) and January (383.3 ng/kg). Elevated AFM1 levels were found in summer in only four samples from the western and other regions, and two samples in the eastern region. This can be attributed to localized and random usage of contaminated feed for dairy cows in those regions. The much lower incidence of elevated AFM1 in comparison to a previous study showed that the outbreak of the crisis due to elevated AFM1 levels in 2013 resulted in a more careful approach to the control of supplementary feedstuff for lactating cows.
•AFM1 concentrations in milk were monitored during one year period in Croatia.•During autumn 9.32% samples exceeding the EU MRL in the eastern region.•In other seasons there were no differences in AFM1 levels between three regions.•Only a few samples with elevated concentrations were found during the summer.
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•One of the highest prevalence of dleptospirosis in Europe with human incidence rate 1.53/100000.•Increased risk for human infections was related to defined parameters.•Variations in ...seroprevalence and changing trends in prevailing serogroups were recorded.•Importance of continuous monitoring with constant multidisciplinary communication.
Leptospirosis is a geographically widespread and globally underestimated zoonosis that affects humans and variety of animals. To identify trends and possible risk factors, joined medical and veterinary teams investigated epidemiology and epizootiology of leptospirosis in Croatia. Retrospective analysis of data obtained from referent diagnostic laboratories included a total of 1917 human and 123964 animal sera tested in the period from 2009 to 2014. We found high human leptospirosis average incidence rate of 1.53/100000 with clear predominance of male patients older than 40 years (sex ratio M/F:3.2; median age 51±15.1years). Statistical analysis revealed seasonal and annual variations of incidence in humans that were primarily associated with favourable weather conditions (temperature 10–19, 9°C and precipitation above 100mm/m2). Majority of infections in humans were caused by serogroups Sejroe, Australis and Icterohaemorrhagiae. Notable variations in seroprevalence and changing trends in prevailing serogroups were recorded in most of the domestic animals and during the entire period of investigation. All of the observed findings underline leptospirosis as a significant human and veterinary public health threat and emphasize the importance of continuous multidisciplinary surveillance. We also argue that only input from both professions improves our overall knowledge on leptospirosis and leads to better and more efficient prevention and control strategies.
This study was conducted to evaluate withdrawal time of levamisole in eggs after oral administration in laying hens at different doses. Sampling of eggs was conducted for 37 days after the end of ...treatment, and levamisole concentrations were measured by liquid chromatography-tandem mass spectrometry validated according to the Commission Decision 2002/657/EC. Estimated validation parameters were as follows: decision limit, 0.54 μg/kg; detection capability, 0.56 μg/kg; limit of detection, 0.04 μg/kg; limit of quantification, 0.15 μg/kg; accuracy (recovery), between 92.9 and 102.3%; precision (relative standard deviation), ≤4.62%; and within-laboratory precision (relative standard deviation), ≤5.19%. Levamisole residue levels were significantly higher in egg yolks than in egg whites. The highest levels of levamisole were detected on day 2 posttreatment in groups receiving 50 mg/kg of body weight (556.2 μg/kg in egg yolks and 166.5 μg/kg in egg whites). Significant elimination occurred within 5 days after the cessation of treatment in all groups, with an elimination half-life of 1.3 days. Levamisole was still detectable on day 30 after the end of treatment in egg whites (0.06 μg/kg) and on day 37 in egg yolks (0.06 μg/kg). The longest withdrawal time for levamisole in eggs (14.9 days) was determined in a group treated with 25 mg of levamisole per kg of body weight for two consecutive days. According to the results, oral treatment of laying hens with levamisole may result in noncompliant egg samples even 14 days after treatment.
•Bosnia and Herzegovina is an endemic for brucellosis in Southeastern Europe.•B. melitensis shows low rates of resistance to commonly used antimicrobials.•A high rate of resistance was observed with ...azithromycin in all three broth media.•The high rate of Brucella resistance to trimethoprim-sulfamethoxazole (T/S) is abolished in alternative broths.•Thymine- and thymidine-depleted broths should be used to test susceptibility for T/S.
Brucellosis is a ubiquitous emergent bacterial zoonotic disease causing significant human morbidity in Bosnia and Herzegovina. So far, a high rate of resistant Brucella has been found worldwide. This study prospectively analysed the rates of resistance among human Brucella melitensis strains isolated in Bosnia and Herzegovina.
This study included 108 B. melitensis isolates from 209 patients diagnosed at five medical centres in Bosnia and Herzegovina. The resistance profiles of the B. melitensis isolates for the 13 most commonly used antimicrobials were studied in standard Brucella broth (BB) and cation-adjusted Mueller-Hinton broth (CAMHB) supplemented with 4% lysed horse blood or 5% defibrinated sheep blood.
Of the 209 patients, B. melitensis blood cultures were positive for 111 (53.1%). Among the 108 isolates investigated, 91 (84.3%) were resistant to trimethoprim-sulfamethoxazole on BB, but not on either CAMHB. Nearly all isolates (>90%) were resistant to azithromycin on BB and both CAMHBs.
We observed a high rate of B. melitensis resistance to azithromycin. The high rate of resistance to trimethoprim-sulfamethoxazole that we observed was related to BB, so an alternative broth should be used, such as the enriched CAMHBs in this study, for evaluating resistance to trimethoprim-sulfamethoxazole. Whole-genome sequencing studies are needed to understand the development of antimicrobial resistance in B. melitensis strains isolated from humans.
Aflatoxin M1 (AFM1) concentrations were determined in raw and UHT cow milk samples collected in different regions of Bosnia and Herzegovina and Croatia during the autumn months of 2014. The mean AFM1 ...levels in the raw milk samples were (ng/kg): 6.22 in Bosnia and Herzegovina, 5.65 in Croatia. In all except one milk sample, AMF1 levels were below the LOQ value of 34.2 ng/kg (ELISA method). In four milk samples, AFM1 concentrations exceeded the EU MRL of 50 ng/kg. Samples were subjected to LC-MS/MS analysis which confirmed elevated values determined by ELISA. Elevated levels were in the range 56.6–132.6 ng/kg. Two positive milk samples from Bosnia and Herzegovina originated from Una Sana Canton, two from Croatia from eastern Croatia. The highest AFM1 levels of 132.6 ng/kg was measured in milk from eastern Croatia. In 214 samples of processed UHT milk from Bosnia and Herzegovina and Croatia, AFM1 ranged from 2.29 ng/kg to 21.4 ng/kg, all below the LOQ value. AFM1 exceeded the EU MRL value in only 0.62% of milk samples, indicating the sporadic use of contaminated feedstuff at farms in both countries.
•Raw and UHT milk were collected in Bosnia and Herzegovina and Croatia.•During autumn 2014 only 0.62% raw milk exceeded the maximum EU limit for AFM1.•Two samples from Bosnia and Herzegovina and two from Croatia exceeded EU limit.•Four elevated AFM1 concentrations were confirmed by LC-MS/MS.•In all UHT milk samples AFM1 levels were below LOQ of the ELISA method.
Brucellosis is an emergent and endemic zoonotic disease in Bosnia and Herzegovina. In this report we have diagnosed the first case of human brucellosis in Bosnia and Herzegovina, using molecular and ...microbiological tests, caused by live attenuated Brucella melitensis Rev.1 strain. The infection was caused through unintentional exposure to vaccination of small ruminants in Bosnia and Herzegovina and without any prior accidental self-injection of vaccine suspension.
Background: Klebsiella pneumoniae is a bacterial species that often causes infections in humans. Infections occur most frequently in hospitalised or immunocompromised patients and are treated with ...antimicrobials. In recent decades, K. pneumoniae has developed significant resistance to many antimicrobials. Objective: The main goal of this study was to determine the frequency of resistance of isolated K. pneumoniae strains from urine samples of hospital patients and outpatients, and to find evidence of ESBL strains and their resistance to certain antibiotics. Methods: During the study period, Klebsiella pneumonia was isolated from the urine samples of 430 patients. The procedure for processing of urine samples, identification, susceptibility toward antimicrobials and evidence of ESBL strains were carried out according to the recommended standards. Results: Of the total K. pneumoniae isolates, 153 (35.6%) were isolated from hospital patients and 277 (64.4%) from outpatients. Strains isolated from hospital patients were resistant to each tested antibiotic. ESBL strains were detected in 169 (39.30%) samples, 92 (60.13%) from hospital patients and 77 (27.8%) from outpatients. Conclusion: Strains of K. pneumoniae isolated from the urine of hospital patients and outpatients have developed significant resistance against all tested antibiotic substances. A higher occurrence of ESBL strains was observed in hospital patients than in outpatients. ESBL strains were resistant to all penicillins and almost all cephalosporins. Highly effective antimicrobials were amikacin, colistine, carbapenem and fosfomycin. The best therapeutic results were achieved when patients were treated with fosfomycin and imipenem.
Epidemija španjolske gripe smatra se najtežom i najopasnijom pandemijom modernog vremena. Nijedna bolest, rat, glad ili drugi događaj u povijesti ljudskoga roda nije prouzročio toliko smrtnosti ljudi ...u tako kratkom vremenu. Započela je u proljeće 1918. godine i zahvatila je čitavi svijet. Po svemu sudeći bila je najveća prirodna katastrofa početkom XX. stoljeća. Prvi pandemijski val započeo je u proljeće 1918. godine (Kansas, SAD) kao blagi oblik bolesti, smrtni slučajevi bili su rijetkost pa zbog toga nije izazvao naročitu pozornost. Drugi val je bio iznimno smrtonosan, a krenuo je u jesen 1918. godine. Oboljelo je više od 30% svjetskog pučanstva (oko 500 milijuna ljudi), a smatra se da je umrlo od 50 do 100 milijuna ljudi. Unatoč toga španjolska gripa je nestala iz povijesti. Bila je zasjenjena Prvim svjetskim ratom i njegovim vojnim te društveno političkim posljedicama. Španjolska gripa može se smatrati „povijesnom nezgodom” i još jednom okrutnom posljedicom strašnog Prvog svjetskog rata.
Brucellosis in swine was surveyed from 2011 to 2015 in 13 counties in Croatia. A total of 3230 breeding males were tested serologically, and positive reactions were confirmed in 42 (1.3%) males from ...17 farms. A total of 641 sows with abortion or reproductive problems were tested, and positive reactions were confirmed in 34 (5.3%). Organs from 68 swine were tested for bacteria, and Brucella spp. was isolated from 47 (69.1%). B. suis was identified in 45 isolates from domestic swine and 2 isolates from wild boar in six counties in Croatia, and all isolates were found to be B. suis biovar 2 based on Bru-up/Bru-low, Bruce-ladder, Suis-ladder and RFLP-based PCR assays. These results indicate that brucellosis is difficult to eradicate in free-range and semi-free-range swine farming, particularly in areas where contact with wild boar is possible. Further disease control measures are required.Key words: Brucella suis biovar 2; swine; abortion; prevalence; CroatiaBRUCELOZA PRAŠIČEV, POVZROČENA Z BAKTERIJO Brucella suis BIOVAR 2 NA HRVAŠKEMBrucelozo pri prašičih smo spremljali od leta 2011 do leta 2015 v 13 hrvaških občinah. S serološko analizo smo preverili prisotnost bruceloze pri 3230 samcih v razplodu. Pozitivne reakcije smo ugotovili pri skupno 42 merjascih s 17 farm, kar predstavlja 1,3 % živali. S serološkimi testi smo preverili prisotnost protiteles proti bruceli tudi pri 641 plemenskih svinjah, ki so zvrgle ali imele težave z zabrejitvijo. Pozitivna reakcija je bila ugotovljena pri 34 svinjah, kar predstavlja 5,3 % vseh testiranih živali. Notranje organe 68 svinj iz šestih občin smo uporabili za osamitev bakterij Brucella spp. Bakterije smo ugotovili pri 47 vzorcih (69,1 %). Bakterijo Brucella suis smo odkrili v vseh 47 vzorcih, izmed katerih jih je bilo 45 od domačih plemenskih svinj, dva vzorca pa sta bila od divjih svinj. Vse izolirane bakterije so pripadale sevu B. suis biovar 2, kot so pokazale dodatne analize z uporabo metod Bruup/Bru-low, Bruce-ladder, Suis-ladder in RFLP. Ti rezultati kažejo, da je popolno izkoreninjenje bruceloze težavno, še posebej v prostih rejah prašičev, kjer obstajajo možnosti stika z divjimi prašiči. Zato bi bilo v prihodnje potrebno razmisliti o dodatnih načinih nadzora nad to nevarno boleznijo prašičev.Ključne besede: Brucella suis biovar 2; prašiči; zvrg; pojavnost; Hrvaška
Hepatitis E is becoming a growing health concern in European countries as an increase of sporadic human cases of unknown origin has been recorded lately. Its causative agent, Hepatitis E virus (HEV), ...is known to have zoonotic potential and thus the role of domestic and wild animals in the chain of viral spread should be considered when investigating risk factors and the epidemiology of the disease. A comprehensive survey based on viral RNA detection was carried out in Croatia including blood, spleen and liver samples originating from 1816 different domestic and wild animals and digestive gland samples from 538 molluscs. A high HEV prevalence was detected in domestic pigs (24.5 %) and wild boars (12.3 %), whereas cattle, molluscs, ruminant and carnivore wildlife samples tested negative. Molecular characterization of both ORF1 and ORF2 genomic regions confirmed the phylogenetic clustering of the obtained sequences into genotype 3, previously reported in Europe. Furthermore, our results proved the presence of identical sequence variants in different samples, regardless of their origin, age or habitat of the host, suggesting transmission events between domestic swine, as well as between domestic swine and wild boars in the country. Moreover, a close genetic relationship of Croatian animal strains and known human HEV strains from GenBank opens the question of possible cross-species HEV transmission in Croatia, especially in the areas with an intensive swine production.
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