Croatian Veterinary Institute Zagreb

Zagreb, Croatia

Croatian Veterinary Institute Zagreb

Zagreb, Croatia
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Lugomer M.D.,Croatian Veterinary Institute Zagreb | Pavlicek D.,Croatian Veterinary Institute Zagreb | Bilandzic N.,Croatian Veterinary Institute Zagreb | Majnaric D.,Croatian Veterinary Institute Zagreb
Mljekarstvo | Year: 2017

A new analytical method using high performance liquid chromatography-diode array detector (HPLC-DAD) was developed for the analysis of 18 benzimidazoles and their metabolites in milk. Samples were extracted with acetonitrile and n-hexane and purified by polymer cation exchange (PCX) solid phase extraction cartridges. LC separation was performed on Xbridge C18 with gradient elution using acetonitrile and ammonium acetate buffer. The DAD detection was set at 298, 312, 254 and 290 nm. The method was validated according to the criteria of Commission Decision 2002/657/EC. The following validation parameters were set: accuracy (expressed as recovery) 31.7-137.6 %, limit of decision (CCα) 6.0-120.6 µg kg-1, detection capability (CCβ) 6.1-120.8 µg kg-1, limit of detection (LOD) 1-4 µg kg-1, limit of quantification (LOQ) 4-18 µg kg-1, precision as CV 7.0-22.5 %, withinlaboratory reproducibility expressed as CV 8.8-30.6 %. Finally, the developed method was applied to the analysis of collected milk samples. A total of 50 milk samples was analysed for benzimidazole residues. All obtained concentrations for all compounds were below the LOQ values. © 2017, Hrvatska Mljekarska Udruga. All rights reserved.


Pleadin J.,Croatian Veterinary Institute Zagreb | Frece J.,University of Zagreb | Lesic T.,Croatian Veterinary Institute Zagreb | Zadravec M.,Croatian Veterinary Institute Zagreb | And 3 more authors.
Food Additives and Contaminants: Part B Surveillance | Year: 2017

The aim of this study was to investigate the occurrence of deoxynivalenol (DON) and zearalenone (ZEN) in unprocessed cereals and soybean sampled in 2014 and 2015 from different fields located in Croatian regions. A total of 306 samples were analysed for DON and 415 samples for ZEN concentrations using quantitative ELISA methods. In 2014, DON and ZEN were determined in all samples in the mean concentrations of 1,461 ± 2,265 µg/kg and 656 ± 853 µg/kg, respectively, while in 2015 these means were 2,687 ± 2,731 µg/kg and 1,140 ± 1,630 µg/kg, respectively. The cultivation year significantly (p < 0.05) influenced mycotoxin concentrations, whereas the influence of cultivation region was seen with ZEN for all cereals except soybean, and not seen with DON at all. A higher contamination determined during 2015 could be explained by high to extreme humidity evidenced in the period of cereals’ growth and harvesting. © 2017 Informa UK Limited, trading as Taylor & Francis Group


PubMed | Croatian Veterinary Institute Zagreb, e Veterinary Institute Vinkovci, University of Zagreb and University of Mostar
Type: Journal Article | Journal: Food additives & contaminants. Part B, Surveillance | Year: 2016

The aim of this study was to investigate the occurrence of citrinin in different cereals (n=341) and feedstuffs (n=67) coming from farms and feed factories established in Croatia and Bosnia & Herzegovina. The highest mean citrinin concentration (103102g/kg) was observed in cereals sampled in Bosnia & Herzegovina during 2014, although significantly different levels between the two countries were not observed. Across the cereal samples, the maximal citrinin concentration was determined in wheat (429g/kg), while across the feedstuffs, the highest concentration was found in pig feed (63g/kg). Despite of the increased citrinin levels found in some samples, especially wheat, the obtained values cannot be compared against the maximum limits, since no such levels are stipulated under the applicable legislation. But, given that data on citrinin are very scarce, they can serve as an indicator of cereal and feed contamination in this part of Europe.


Persi N.,Croatian Veterinary Institute Zagreb | Pleadin J.,Croatian Veterinary Institute Zagreb | Kovacevic D.,Josip Juraj Strossmayer University of Osijek | Scortichini G.,Instituto Zooprofilattico Sperimentale Dellabruzzo E Del Molise G Caporale | Milone S.,Instituto Zooprofilattico Sperimentale Dellabruzzo E Del Molise G Caporale
Meat Science | Year: 2014

The aim of this study was to determine ochratoxin A (OTA) concentrations in the raw materials and cooked meat products made from pigs sub-chronically exposed to OTA. The treated animal group (n. = 5) was administered with 300. μg OTA/kg of feed for 30. days, whereas the control group (n. = 5) was left untreated. OTA concentrations were quantified using immunoassay (ELISA) and high performance liquid chromatography with fluorescence detection (HPLC-FD). OTA concentration was the highest in the kidney, followed by the lungs, liver, blood, spleen, heart, and adipose tissue. As for the final meat products, the highest average OTA concentration was detected in black pudding sausages (14.02. ±. 2.75. μg/kg), then in liver sausages (13.77. ±. 3.92. μg/kg), while the lowest was found in pâté (9.33. ±. 2.66. μg/kg). The results pointed out that a sub-chronic pig exposure leads to the accumulation of OTA in raw materials and consequently in meat products, whose level of contamination is directly dependent on OTA contents in raw materials used for their production. © 2013 Elsevier Ltd.


Lesic T.,Croatian Veterinary Institute Zagreb | Pleadin J.,Croatian Veterinary Institute Zagreb | Kresic G.,University of Rijeka | Vahcic N.,University of Zagreb | And 3 more authors.
Journal of Food Composition and Analysis | Year: 2016

In this study, the basic chemical and fatty acid compositions of autochthonous Croatian cow and sheep cheeses in a lamb skin sack (local name: "sir iz mišine") were determined. Also, the influences of ripening period and different starter cultures on chemical and fatty acid composition of these cheeses were investigated. Samples of cow ( n= 20) and sheep ( n= 20) milk cheeses were produced in three different ways: from raw milk without the addition of a starter culture, from pasteurized milk with commercial starter cultures and with previously isolated autochthonous starter cultures ( Lactococcus lactis S1 or Lactobacillus plantarum B or a mixture of both). Samples were taken during a 45-day ripening period (on days 0, 15, 30 and 45). The ripening time significantly affected all basic chemical parameters, while different starter cultures significantly ( p< 0.05) influenced protein, fat and ash content. Ripening time had no significant effect on the representation of the investigated fatty acid groups ( p> 0.05), but a significant difference was found depending on the starter cultures used and the type of cheese analysed, in terms of statistically higher proportion ( p< 0.05) of polyunsaturated fatty acids in the finished sheep's milk cheese (2.58-2.97%) in comparison to the cow's milk cheese (1.93-2.14%). Fatty acids most represented in the analysed cheeses were palmitic, oleic and stearic acid. © 2015 Elsevier Inc.


Domijan A.-M.,University of Zagreb | Pleadin J.,Croatian Veterinary Institute Zagreb | Mihaljevic B.,Ruder Boskovic Institute | Vahcic N.,University of Zagreb | And 2 more authors.
Food Additives and Contaminants - Part A Chemistry, Analysis, Control, Exposure and Risk Assessment | Year: 2015

This study investigated the efficiency of gamma (γ)-irradiation in the reduction of ochratoxin A (OTA) present in dry-cured meat products prepared from intentionally contaminated raw materials from OTA-treated pigs. OTA concentrations determined in the samples (n = 24) ranged from 25.8 μg kg–1 in bacon to 17.8 μg kg–1 in smoked ham. After γ-irradiation at doses of 3, 7 and 10 kGy (i.e. the doses used in the food industry), a dose-depended OTA reduction was observed; however, it was not statistically significant. The mean OTA reduction achieved with 3-, 7- and 10-kGy γ-doses was approximated to 8.5%, 13.9% and 22.5%, respectively. The storage of irradiated samples (1 month, 4°C) did not significantly affect OTA levels. Based on the correlation between the OTA reduction level and basic chemical composition of dry-cured meat samples, OTA reduction may be linked to the samples’ fat content. The results indicate that γ-irradiation can reduce OTA levels in dry-cured meat products, but only to a limited extent due to the complexity of the matrix. © 2015 Taylor & Francis.


Pleadin J.,Croatian Veterinary Institute Zagreb | Persi N.,Croatian Veterinary Institute Zagreb | Kovacevic D.,Josip Juraj Strossmayer University of Osijek | Vulic A.,Croatian Veterinary Institute Zagreb | And 2 more authors.
Food Additives and Contaminants: Part B Surveillance | Year: 2014

The aim of this study was to investigate the possibilities of ochratoxin A (OTA) reduction in home-made meat products. Meat sausages (n = 50) produced from raw materials coming from pigs exposed to OTA-contaminated feed, were subject to common heat processes practiced in households (cooking, frying and baking). Concentrations of OTA in pre- and post-processed products were quantified using a validated immunoassay method, enzyme-linked immunosorbent assay, and confirmed using a high-performance liquid chromatography with fluorescence detection. In line with the differences in recipes used and the degree of OTA accumulation in raw materials, OTA concentrations established in Mediterranean and roast sausages were lower than those found in liver and blood sausages. Baking of contaminated sausages at the temperatures of 190–220°C (for 60 min) resulted in significant reduction of OTA levels (75.8%), while 30-min cooking (at 100°C) and frying (at 170°C) proved to be significantly less effective (e.g. yielding OTA reductions of 7.4% and 12.6%, respectively). The results pointed out that despite high OTA stability, heat processes are capable of reducing its concentration in home-made meat products, depending on the processing modality used. © 2014, © 2014 Taylor & Francis.


Pleadin J.,Croatian Veterinary Institute Zagreb | Vulic A.,Croatian Veterinary Institute Zagreb | Persi N.,Croatian Veterinary Institute Zagreb | Skrivanko M.,Croatian Veterinary Institute | And 2 more authors.
Food Control | Year: 2014

The aim of the study was to determine the level of aflatoxin B1 (AFB1) in maize sampled from farms and feed factories situated in Northern, Central and Eastern Croatia during 2013, following the occurrence of cow milk AFM1 contamination. Maize samples (n=633) were analysed using Enzyme-Linked Immunosorbent Assay (ELISA) as a screening method and High Performance Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS) as a confirmatory method. Mean AFB1 value found in maize coming from all investigated regions equalled to 81μg/kg, with the maximal value of 2072μg/kg found in maize obtained from Eastern Croatia. The observed contamination might have arisen on the grounds of extremely hot (>98%) and dry (<2%) weather witnessed from May to September 2012 during the maize growth and harvesting period, which might have favoured AFB1 production and consequently the contamination of dairy cattle feeds. In order to prevent the adverse effects of AFB1 on humans and animals, and also to reduce losses in agricultural production, systematic monitoring and further investigations of AFB1 contamination are necessary. © 2013 Elsevier Ltd.


Pleadin J.,Croatian Veterinary Institute Zagreb | Vulic A.,Croatian Veterinary Institute Zagreb | Persi N.,Croatian Veterinary Institute Zagreb | Skrivanko M.,Croatian Veterinary Institute | And 2 more authors.
Food Control | Year: 2015

The aim of the study was to investigate annual and regional differences in the level of aflatoxin B1 (AFB1) in grains and dairy cattle feed. Maize (n=972), wheat (n=201), barley (n=147), oat (n=136), grain mixtures (n=168), and dairy cattle feed (n=325) were sampled from 2009 to 2013 on different farms and in different farm factories situated in four Croatian regions. The samples were analysed for AFB1 using the validated ELISA immunoassay. AFB1 was determined in 16.4% of all investigated samples, among which maize was proven to be the most contaminated, with 21.7% of the samples recovered during 2013 harbouring AFB1 in concentrations over the permissible ones. Levels higher than permitted were observed in 17.9% and 12.3% of grain mixtures and dairy cattle feed, respectively, whereas concentrations of AFB1 determined in other crops throughout the investigated period met the stipulated requirements. The results revealed the AFB1 occurrence to be significantly (p<0.05) dependent on the cultivation region, with the highest levels generally found in maize harvested in 2013 and consequently in grain mixtures and cattle feed that can most likely be associated with climatic conditions as the most critical factor for mould formation, and thus also AFB1 production. © 2014 Elsevier Ltd.


PubMed | Croatian Veterinary Institute Zagreb
Type: Journal Article | Journal: Food additives & contaminants. Part B, Surveillance | Year: 2014

The aim of this study was to investigate the possibilities of ochratoxin A (OTA) reduction in home-made meat products. Meat sausages (n=50) produced from raw materials coming from pigs exposed to OTA-contaminated feed, were subject to common heat processes practiced in households (cooking, frying and baking). Concentrations of OTA in pre- and post-processed products were quantified using a validated immunoassay method, enzyme-linked immunosorbent assay, and confirmed using a high-performance liquid chromatography with fluorescence detection. In line with the differences in recipes used and the degree of OTA accumulation in raw materials, OTA concentrations established in Mediterranean and roast sausages were lower than those found in liver and blood sausages. Baking of contaminated sausages at the temperatures of 190-220C (for 60min) resulted in significant reduction of OTA levels (75.8%), while 30-min cooking (at 100C) and frying (at 170C) proved to be significantly less effective (e.g. yielding OTA reductions of 7.4% and 12.6%, respectively). The results pointed out that despite high OTA stability, heat processes are capable of reducing its concentration in home-made meat products, depending on the processing modality used.

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