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Electrotechnologies gained increased popularity as non-thermal processing of foods as they offer many advantages over conventional thermal processing. These technologies effectively inactivate microorganisms, enhance the processing efficacy at lower treatment intensity (e.g. pressing, pre-treatment for drying, extraction etc.) with preservation of nutritional, functional and sensory characteristics of processed foods. Moreover, these technologies can be applied for targeted and controlled enzymatic treatments. Structural and conformational changes under these treatments may influence enzymatic inactivation, but also activate enzymes or enhance their activity. Therefore, the enzyme characteristics and treatment conditions could be finely tuned toward desirable properties and effects in food production and processing.

The aroma of a cheese is one of the key quality parameters and signs of distinguishing certain cheese types by consumers. Therefore, the aim of this paper is review the existing knowledge of the aromatic profile of cheeses ripening in an animal skin. Cheeses that ripen in an animal skin and produced in similar technology in Croatia (mišina cheese in a sack), in Bosnia and Herzegovina (Cheese in a sack (local name: mjeh), Montenegro (Cheese in a sack (local name: mjeh), Turkey (Tulum cheese), Lebanon (Darfiyeh cheese) and eastern Algeria (Bouhezza cheese). The specific characteristic of these cheeses is ripening in an animal skin, as in a sack (of ewe or goat). The aromatic profile of these cheeses has not been sufficiently investigated and has been conducted primarily on Tulum and to a smaller extent of Lebanese Darfiyeh cheese, and cheese in a sack. There is no data about other cheeses that ripen in an animal skin. Since very little research has been made on the aromatic profile of these cheeses, further research should be conducted., Aroma sira jedan je od ključnih parametara kvalitete i znakova raspoznavanja pojedine vrste sira od strane potrošača. Stoga je cilj ovog rada pregled postojećeg znanja o aromatskom profilu sireva, koji se proizvode na tradicionalan način i zriju u životinjskoj koži. Sirevi, koji zriju u životinjskoj koži i imaju sličnu tehnologiju, proizvode se u Hrvatskoj (Sir iz mišine), na području Bosne i Hercegovine (Sir iz mijeha), Crne Gore (Sir iz mijeha), Turske (Tulum sir), Libanona (Darfiyeh) i istočnog Alžira (Bouhezza sir). Specifičnost ovih sireva je zrenje u životinjskoj koži, odnosno mišini (ovčjoj, kozjoj). Aromatski profil navedenih sireva je slabo istražen, a istraživanja su najvećim dijelom provedena na Tulum siru i manjim dijelom na libanonskom Darfiyeh siru, siru iz mišine, dok za ostale sireve,koji zriju u životinjskoj koži nema nikakvih podataka. Obzirom da je vrlo malo istraživanja o aromatskom profilu ovih sireva do sada napravljeno, potrebno je provoditi daljnja istraživanja u ovom pravcu.

This study focused on monitoring changes in aflatoxin M1 (AFM1) concentrations during produc- tion and storage of different fermented milks using selected probiotic and nonprobiotic combined cultures. Milk samples intended for fermentation were intentionally contaminated by adding a stan- dard of AFM1. All of the tested cultures caused remarkable reductions in AFM1 concentrations during the fermentation process. Probiotic cultures were more effective than nonprobiotic cultures, with Lactobacillus casei LC-01 strain being the most efficient, achieving a reduction level of approximately 58%. Among the nonprobiotic cultures, yoghurt culture YC- 380 was the most effi- cient, with a reduction level of approximately 41%.

The aim of this study was to investigate into 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 analyzed 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.

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 (103 ± 102 µg/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 (429 µg/kg), while across the feedstuffs, the highest concentration was found in pig feed (63 µg/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.

Traditionally, equid milk has been used as an alternative to human milk for infants or children mainly because of its similarities in nutrient composition, hypoallergenicity, immune modulation and antimicrobial activity. As a highly nutritious substrate, it possesses great potential for probiotic bacteria isolation. The aim of this study was to assess the probiotic potential of Lactobacillus plantarum M2 and L. plantarum KO9 isolated from donkey and mare milk, respectively, and determine TNF-α suppression attributed to their extracellular metabolites in lipopolysaccharide- (LPS-)stimulated human peripheral blood mononuclear cells (PBMCs). Resistance to simulated gastric and intestinal conditions, antimicrobial activity against pathogens, autoaggregation, coaggregation, biofilm quantification, antioxidative potential and haemolytic activity were determined. The isolated strains demonstrated great probiotic potential and their extracellular metabolites with molecular mass smaller than 2000 Da supressed TNF-α production up to 67% in LPS-stimulated PBMCs exerting high anti-inflammatory activity. Extracellular metabolites did not show any cyto/genotoxic effects toward PBMCs.

Toxicity of gamma irradiated mycotoxins aflatoxin B1 (AFB1) and ochratoxin A (OTA) was investigated in vitro. AFB1 and OTA stock solutions (50 mM, in methanol) were gamma irradiated (5 and 10 kGy) and non-irradiated and irradiated mycotoxins solutions were tested for cytotoxicity on Pk15, HepG2 and SH-SY5Y cell lines (MTT assay, 1–500 lM concentration range ; 24 h exposure). Degradation of mycotoxin molecules was examined by liquid chromatography tandem mass spectrometry (HPLC-MS/MS). AFB1 and OTA radiolytic products were less toxic than the parent mycotoxins to all of the tested cell lines. Gamma irradiation even at 5 kGy had effect on AFB1 and OTA molecules however, this effect was dependent on chemical structure of mycotoxin. Since gamma irradiation at low dose reduced initial level of both mycotoxins, and gamma irradiated mycotoxins had lower toxicity in comparison to non-irradiated mycotoxins, it can be concluded that gamma irradiation could be used as decontamination method.

The aim of this study was to investigate ochratoxin A (OTA) and citrinin (CIT) co- occurrence in different unprocessed cereals (n=189) originating from Croatia during a three- year investigation period (2014-2016) using validated enzyme immunoassay (ELISA) methods. CIT and OTA were determined in 49% and 7% of samples, respectively. Significantly higher (p < 0.05) overall mean concentrations were determined for CIT (66.8 ± 76.0 µg/kg) in comparison to OTA (5.2 ± 1.1 µg/kg). Based on the analysis of all investigated cereals, CIT was found about 15 times more frequently than OTA and in similarly (15-fold) higher concentrations, irrespective of the cultivation year. The results revealed a moderately positive correlation between OTA and CIT concentrations in maize (rs = 0.44) and wheat (rs = 0.59), whereas in barley and oat this correlation (p > 0.01) was not significant.

In this paper the effect of aflatoxin B1, ochratoxin A and zearalenon on morphology, growth parameters and metabolic activity of yeasts Saccharomyces cerevisiae, Saccharomyces uvarum, Candida utilis and Kluyveromyces marxianus was determined. The results showed that the three mycotoxins affected the morphology of all these yeasts, primarily the cell diameter, but not their final cell count. Fourier transform infrared spectroscopy showed that the yeast membranes bound the mycotoxins, C. utilis in particular. The cell membranes of most yeasts underwent denaturation, except S. uvarum exposed to ochratoxin A and zearalenone. In the early stage of fermentation, all mycotoxin-exposed yeasts had lower metabolic activity and biomass growth than controls, but fermentation products and biomass concentrations reached the control levels by the end of the fermentation, except for C. utilis exposed to 20 μg/mL of zearalenone. The adaptive response to mycotoxins suggests that certain yeasts could be used to control mycotoxin concentrations in the production of fermented food and beverages.

Mycotoxin patulin is one of the quality indicators for apple juice. Like other mycotoxins, it raises consumer health concerns. The issue of low quality is particularly relevant for apples provided by small producers, whose quality control may not be standardised. As sulphur dioxide (SO2) is common in fruit preservation against fungi, the aim of this study was to determine how efficient it is in degrading patulin in apple juices stored in real-life conditions. This included refrigerated (4 °C) and non-refrigerated warehouses/environments (30 °C) over 8, 10, 12, and 20 weeks of storage. Apple juice was diluted to 0.010 μg g-1, 0.050 μg g-1, and 0.100 μg g-1 of patulin. SO2 was added to each sample in the amounts of 250 μg mL-1 and 50 μg mL-1. Untreated juice samples for each patulin concentration served as controls under the same experimental conditions. Patulin content was determined with high performance liquid chromatography. The best degradation was observed with 250 μg mL-1 of SO2 at 30 °C regardless of the patulin baseline concentration. Although treatment with SO2 and refrigeration did not fully remove patulin, it was highly efficient over twelve weeks of storage. Our results suggest that patulin levels can be reduced between 33 and 100 % at 30 °C and up to 100 % at 4 °C., Mikotoksin patulin jedan je od pokazatelja kakvoće jabučnog soka. Poput drugih mikotoksina, prisutnost patulina predstavlja zdravstveni problem. Pitanje loše kakvoće jabuka odnosi se na mala obiteljska gospodarstva, gdje se kakvoća korištenih jabuka ne može standardizirati. Prisutnost plijesni i mikotoksina zdravstveni je problem, a SO2 pospješuje razgradnju patulina. Svrha ove studije bila je odrediti razgradnja patulina u jabučnom soku tretiranim sa SO2 pri različitim temperaturama skladištenja. Utjecaj SO2 na razgradnju patulina ispitivana je u uzorcima pohranjenima u svakodnevnim uvjetima. To uključuje hladnjak (4 °C) i nehlađena skladišta/okoliš (30 °C) tijekom 8, 10, 12 i 20 tjedana. Jabučni sok razrijeđen je na koncentracije 0,010 μg g-1, 0,050 μg g-1 i 0,100 μg g-1 patulina. U svaki uzorak dodan je SO2 u količini od 250 μg mL-1 i 50 μg mL-1. Uzorci soka u koji nije dodan SO2 za svaku koncentracijsku razinu patulina poslužili su kao kontrola i bili su pod istim eksperimentalnim uvjetima. Patulin u uzorcima određivan je pomoću tekućinskog kromatografa visoke djelotvornosti (HPLC). Najveća razgradnja patulina zabilježena je u termostatskim uvjetima s koncentracijom SO2 od 250 μg mL-1, bez obzira na koncentraciju patulina kontrolnog uzorka. Iako hlađeni uvjeti i dodatak SO2 nisu potpuno uklonili patulin iz uzoraka, nakon dvanaest tjedana skladištenja uočena je visoka učinkovitost razgradnje. Rezultati pokazuju da se razine patulina mogu smanjiti 33-100 % pri temperaturi od 30 °C i 0-100 % pri 4 °C.

In this study, the occurrence of Fusarium mycotoxins deoxynivalenol (DON), zearalenone (ZEA) and fumonisins (FUM) was investigated in a total of 257 samples of unprocessed cereals (maize, wheat and barley), sampled annually in Bosnia and Herzegovina throughout the 2013-2015 harvesting period. The levels of mycotoxins were determined using a validated ELISA method. The results showed maize to be the most contaminated cereal among the three, with DON detected in 85%, ZEA in 73% and FUM in 67% of samples, the mean concentrations of the toxins observed during the study period thereby being 984±957 μg/kg, 326±314 μg/kg, and 1, 259±1, 161 μg/kg, respectively. Twenty samples (7.8%) were proven inadmissible for consumption as foodstuffs (Commission Regulation 1881/2006), and 3 samples (1.2%) were proven inadmissible even as feedstuffs (Commission Recommendation 2006/576/EC). Significantly higher (p < 0.05) mycotoxin levels determined in samples harvested in 2014 as compared to those harvested in 2013 and 2015 could be associated with heavy rainfall periods witnessed in 2014 that could favour the formation of moulds, and consequently also the increased production of Fusarium mycotoxins.

In this study, the levels of aflatoxin B1 (AFB1), ochratoxin A (OTA), zearalenone (ZEN), deoxynivalenol (DON) and fumonisins (FUM) in unprocessed cereals (n = 189) and cereal-based products (n = 61) were determined using validated ELISA methods. All samples originated from either conventional or organic production corresponded to the 2015 harvest in Croatia. Based on the mean mycotoxin concentrations, the risk for the consumer to exceed the tolerable daily intake (TDI) for these toxins by the consumption of both types of cereals and cereal-based products was assessed. Mycotoxin contamination of organic cereals and organic cereal-based products was not significantly different (p > 0.05). Given that the exposure assessment resulted in a small fraction of the TDI (maximum: DON, 12% of TDI), the levels of the investigated mycotoxins in both types of cereals and cereal-based products from the 2015 harvest did not pose a human health hazard.

In this study, the effect of an isolated and well-characterized autochthonous starter culture (ASC), various sugars (glucose and maltodextrin, w = 0.8 %) and different fermentation temperatures (12 and 20 oC) on fermentation and quality of Slavonian Kulen produced using the traditional technology and recipe were investigated. Physicochemical and microbiological analyses were carried out after 20 days of fermentation. Upon the completion of the production process (90 days), a sensory analysis was carried out, too. Furthermore, pH valuewas continuously measured throughout the twenty-day fermentation period. The addition of an ASC and sugars and fermentation temperature variations significantly (P < 0.05) affected the instrumental colour and texture parameters of the Slavonian Kulen. The fermentation process was most intense in the samples supplemented with an ASC and 0.8 % - glucose and fermented at 20 °C. Microbiological analysis showed samples supplemented with an ASC and fermented at higher temperatures to contain a higher number of lactic acid bacteria (LABs) and coagulase- negative staphylococci (CNS) and to be safe. Sensorial evaluation confirmed the outcomes of physicochemical and microbiological analyses and showed differences between samples fermented at different temperatures and supplemented with sugars and an autochthonous starter culture.

The purpose of this study was to investigate the effect of non-thermal technology, high power ultrasound (HPU) on inactivation of Aspergillus ochraceus 318, Penicillium expansum 565, Rhodotorula sp. 74, Saccharomyces cerevisiae 5 and Alicyclobacillus acidoterrestris DSM 3922 in clear juices and nectars from apple, blueberry and cranberry juice concentrate. Inoculated juice and nectars were treated by high power ultrasound (20 kHz) according to procedure set by central composite design (CCD). Three operational parameters, amplitude (60, 90 and 120 µm), temperature (20, 40 and 60 °C), and treatment time (3, 6 or 9 min) were varied in order to observe the influence of ultrasound and combination of ultrasound and slight heating (thermosonication) on growth and inactivation of selected microorganisms. Number of vegetative cells of A. acidoterrestris DSM 3922 were not significantly reduced by high power ultrasound (p > 0.05), except in apple juice, where statistical significant (p < 0.05) influence of quadratic interaction of amplitude on bacteria reduction were observed. In all samples of fruit juices and nectars in terms of ultrasonic treatment at 60 °C and times of 3, 6 and 9 min, regardless of the value of the amplitude, complete inactivation of the growth of yeasts and moulds were achieved, while at 20 and 40 °C it is not observed. The value of reduction of cells of selected yeasts and moulds for ultrasound treatments at 60 °C and the duration of the 3, 6 and 9 min ranged from 3.556 to 5.934 log units, depending on the initial number of selected yeasts and moulds before treatment.

Urine Ochratoxin A and Sphinganine/Sphingosine Ratio in Residents of the Endemic Nephropathy Area in CroatiaThe most plausible theory of the aetiology of endemic nephropathy links it with exposure to nephrotoxic mycotoxin ochratoxin A (OTA). In this study, the concentration of OTA and sphinganine/sphingosine (Sa/So) ratio, the biomarker of another nephrotoxic mycotoxin fumonisin B1 exposure, were analysed in 45 human urine samples collected in the endemic village of Kaniža in Croatia and in 18 samples from control village. Samples were collected twice from the same persons in 2000 and 2005. In both years the frequency of OTA-positive samples was higher in Kaniža (43 % and 18 %, respectively) than in the control village (28 % and 6 %, respectively). OTA concentrations in samples collected in Kaniža were higher in 2000 than in 2005 (p1 at the same time, while in Kaniža four such samples were collected in 2000 and one in 2005.

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.

Svrha je ovoga istraživanja bila identificirati i okarakterizirati potencijalne autohtone funkcionalne starter kulture izolirane iz domaće kobasice proizvedene od konjskog mesa. Dominantnu su mikrofloru u uzorcima kobasica činile bakterije mliječne kiseline (BMK), a zatim mikrokoki. Od bakterija mliječne kiseline prevladavale su vrste Lactococcus lactis ssp. lactis i Lactobacillus plantarum. Vrsta Lactococcus lactis ssp. lactis nije uobičajena u fermentiranim kobasicama, pa smo ju okarakterizirali kao potencijalnu funkcionalnu starter kulturu. Vrsta Lactococcus lactis ssp. lactis proizvela je značajnu količinu mliječne kiseline, a uspješno je rasla pri 12, 18 i 22 ºC, te u prisutnosti 5 % NaCl. Također je dobro podnijela liofilizaciju i simulirane uvjete želučanog soka i soka tankog crijeva. Osim toga, uspješno je suzbila rast patogena i pokazala je dobra adhezijska svojstva in vitro., The aim of this study is to identify and characterise potential autochthonous functional starter cultures in homemade horsemeat sausage. The dominant microflora in the samples of horsemeat sausage were lactic acid bacteria (LAB), followed by micrococci. Among the LAB, Lactococcus lactis ssp. lactis and Lactobacillus plantarum were the dominant species, and since the first is not common in fermented sausages, we characterised it as a potential functional starter culture. Lactococcus lactis ssp. lactis produced a significant amount of lactic acid, displayed good growth capability at 12, 18 and 22 °C, growth in the presence of 5 % NaCl, good viability after lyophilisation and in simulated gastric and small intestinal juice, antimicrobial activity against test pathogens, and good adhesive properties in vitro.