Climate Change and Mycotoxins Trends in Serbia and Croatia: A 15-Year Review.

This review examines the 15-year presence of mycotoxins in food from Serbia and Croatia to provide a comprehensive overview of trends. Encompassing the timeframe from 2009 to 2023, this study integrates data from both countries and investigates climate change patterns. The results from Serbia focus primarily on maize and milk and show a strong dependence of contamination on weather conditions. However, there is limited data on mycotoxins in cereals other than maize, as well as in other food categories. Conversely, Croatia has a broader spectrum of studies, with significant attention given to milk and maize, along with more research on other cereals, meat, and meat products compared to Serbia. Over the investigated 15-year period, both Serbia and Croatia have experienced notable shifts in climate, including fluctuations in temperature, precipitation, and humidity levels. These changes have significantly influenced agriculture, consequently affecting the occurrence of mycotoxins in various food products. The results summarized in this 15-year review indicate the urgent need for further research and action to address mycotoxins contamination in Serbian and Croatian food supply chains. This urgency is further emphasized by the changing climatic conditions and their potential to exacerbate public health and food safety risks associated with mycotoxins.

Aflatoxins in maize, milk and dairy products from Serbia.

This study presents data on the occurrence of aflatoxins B1, B2, G1 and G2 in maize and aflatoxin M1 (AFM1) in milk and dairy products from Serbia in 2022. A total of 100 maize samples were analysed using liquid chromatography tandem mass spectrometry, while 107 cow and goat milk, besides dairy products were tested with an ELISA method. Aflatoxins contaminated 78% of the maize samples, with aflatoxin B1 as the most prevalent aflatoxin, also at the highest determined level. All milk samples tested positive for AFM1, while contamination in dairy products ranged from 14% to 100%. The hot and dry weather during the 2022 maize growing season, characterised by moderate to extreme drought conditions, significantly increased maize contamination with aflatoxins, impacting subsequent milk and dairy products contamination. The findings of this study confirm the continuous trend and persistent challenge in Serbia concerning the prevalence of aflatoxins in maize and milk, closely linked to climate change.

Sour Cherry Pomace Valorization as a Bakery Fruit Filling: Chemical Composition, Bioactivity, Quality and Sensory Properties.

Sour cherry pomace filling (SCPF) and commercial sour cherry filling (CSCF) produced on a semi-industrial scale were tested and compared in terms of food safety, chemical composition, bioactivity, quality, sensory properties and thermal stability. Both samples were safe for human consumption, thermally stable and there was a lack of syneresis. SCPF had a significantly higher fiber concentration (3.79 g/100 g) due to higher skin fraction and is considered a "source of fibers". The higher skin fraction in SCPF also resulted in a higher mineral quantity (Fe-3.83 mg/kg fw) in comparison to CSCF (Fe-2.87 mg/kg fw). Anthocyanins concentration was lower in SCPF (7.58 mg CGE/100 g fw), suggesting that a significant amount of anthocyanins was removed from SC skin during juice extraction. However, there was a lack of statistical differences in antioxidant activity between the two fillings. CSCF was more spreadable, not as firm and less sticky, with lower storage and loss modulus values than SCPF. However, both fillings exhibited acceptable rheological and textural behaviour for fruit fillings. According to the consumer pastry test, 28 participants preferred each pastry; thus, there was a lack of preference toward any of the tested samples. SCP could be used as a raw material for the bakery fruit fillings industry, which leads to the valorization of food industry by-products.

Microbiological Quality of Raw Donkey Milk from Serbia and Its Antibacterial Properties at Pre-Cooling Temperature.

The aim of this study was to examine the microbiological quality of raw donkey milk of an indigenous Serbian breed as well as the changes in the microbial populations during storage at 4 °C. In addition, antibacterial activity of donkey milk against E. coli, L. monocytogenes and S. aureus at 15 °C as well as the content of the two main antibacterial proteins lysozyme and lactoferrin were investigated. Microbiological examination of 137 individual milk samples collected over a period of 21 months showed good microbiological quality since foodborne pathogens such as Salmonella spp. and L. monocytogenes were not detected in any of the analyzed samples, while the number of E. coli, Enterobacteriaceae, total coliform bacteria, sulfite-reducing Clostridia and aerobic sporogenic bacteria was below the limit of quantification (<1 cfu mL-1). During the six-days storage at 4 °C, total bacterial counts and the counts of lactic acid bacteria remained at the initial level while pathogenic bacteria were not detected. The strongest antibacterial activity of the tested milk was observed against E. coli, while S. aureus was the least sensitive to milk antibacterial compounds. Although further research is needed to fully elucidate the antibacterial mechanism and synergistic activity of different compounds in donkey milk, the high content lysozyme (2.63 ± 0.03 g L-1) and lactoferrin (15.48 mg L-1) observed in tested milk could contribute to its strong antibacterial activity and extension of the storage period during which it can be safely consumed.

Potential health benefits of blueberry and raspberry pomace as functional food ingredients: Dietetic intervention study on healthy women volunteers.

The fruit juice industry generates pomace as a valuable by-product especially rich in polyphenols, dietary fibers, vitamins, minerals, and unsaturated fatty acids. In the cookies used in this study, 30% of the gluten-free flour was replaced with dried and ground blueberry and raspberry pomace, rich source of polyphenols, dietary fibers, linoleic and alpha-linolenic acid. In order to examine whether the addition of blueberry and raspberry pomace in cookie formulation can have beneficial effects on certain blood parameters and anthropometric measurements, the designed cookies were tested in 20 healthy, normally fed female subjects, aged 30-50 years (41.35 ± 8.58 years) over four-week dietetic intervention study. Significant changes in the composition of fatty acids serum phospholipids, decrease in LDL-cholesterol level (20.16%), increase in adiponectin level (25.52%) and decrease in ALT and AST values were observed, thus indicating that inclusion of cookies containing blueberry and raspberry dried and ground pomace to usual diet might have positive effects on certain cardiovascular risk factors and liver function indicators.

Antibacterial Potential of Allium ursinum Extract Prepared by the Green Extraction Method.

The antimicrobial activity of Allium ursinum aqueous extract prepared using high pressure extraction was evaluated. Minimal inhibitory concentrations (MIC) and minimal bactericidal concentrations (MBC) of A. ursinum extract for six bacterial pathogens were determined using the broth macrodilution method. Although the A. ursinum extract was shown to be effective toward all investigated foodborne bacteria, its antimicrobial activity depended on its concentration and bacterial strain. Listeria monocytogenes was the most sensitive to antimicrobial activity of A. ursinum extract among all tested pathogens. Accordingly, the lowest MIC and MBC of A. ursinum extract were determined for L. monocytogenes (28 and 29 mg/mL). The tested extract showed a similar antimicrobial potential to other examined bacterial strains (Salmonella Enteritidis, Proteus hauseri, Enterococcus faecalis and two strains of Escherichia coli) with MIC and MBC values at concentrations of 29 and 30 mg/mL, respectively. The dependence of the antimicrobial activity of the A. ursinum extract on the level of contamination of tested pathogens was also observed. The increase in the contamination level caused an intense reduction in antibacterial potential of the A. ursinum extract. The composition of the A. ursinum extract was analyzed and found to be a good source of polyphenols and sulfur compounds. However, considering the applied extraction method and the HPLC analysis of bioactive compounds, the antimicrobial potential may be attributed more to polyphenol content. The obtained results that the extracts have shown toward food pathogens open the possibility of using the tested extracts as natural additives in a variety of food products.

Fiber concentrates from raspberry and blueberry pomace in gluten-free cookie formulation: Effect on dough rheology and cookie baking properties.

Raspberry and blueberry pomace, by-products from fruit juice industry, were dehydrated to obtain fruit fiber concentrates and valorized as ingredients in gluten-free (GF) cookies formulation. The aim was to evaluate the effect of substituting 30% (w/w) of GF flour mixture with different ratios of blueberry/raspberry fiber concentrates (0/30, 15/15, and 30/0) on rheological properties of cookie dough and cookie baking quality. Generally, the incorporation of the fiber concentrates increased the elastic and the viscous moduli thus producing harder doughs and consequently firmer cookies in comparison to the control. Although having similar total, soluble and insoluble fiber content, blueberry fiber concentrates were characterized with higher water absorption capacity and consequently yielded cookies with more rigid dough structure, higher water loss during baking, lower cookie thickness, higher spread ratio, more dense inner structure, and increased cookie hardness than those containing raspberry fiber concentrates. Enrichment of GF cookies with raspberry and blueberry fiber concentrates resulted in products with dietary fiber content higher than 6 g per 100 g, thus making them candidates for bearing "high fiber" nutrition claim. PRACTICAL APPLICATIONS: Currently available GF bakery products mostly lack dietary fibers (DFs) and other important nutrients, because they are usually manufactured from refined flour and/or starches. As the products with long shelf-life, very popular in daily diet of almost all consumer profiles, cookies could represent good matrices for the addition of functional ingredients, such as DFs. In this paper, GF cookies were enriched with the fiber concentrates recovered from blueberry and raspberry pomace. Application of these fiber sources enabled recycling of fruit processing industry by-products into novel, "high fiber" GF product with improved nutritive profile.

Aflatoxins in maize harvested in the Republic of Serbia over the period 2012-2016.

Aflatoxins (AFs) are one of the most frequent contaminants in maize. The Republic of Serbia represents a leader in terms of maize production and exports in Europe. In recent years, Serbia faced climate changes. Therefore, the main objective of this study was to examine AF occurrence in 3000 maize samples collected during 2012-2016. In maize samples from 2012, 2013, 2015, and 2016, AFs were detected in 72.3%, 24.7%, 36.7%, and 5.0% of the samples, in concentration ranges of 1.0-111.2, 1.2-65.2, 1.1-76.2, and 1.3-6.9 µg kg-1, respectively. Contrary to this, in maize samples from 2014, no AFs were detected. Analysis of weather conditions indicates that 2012 was characterised by extreme drought conditions, 2013 and 2015 by lack of precipitation and high air temperatures, 2014 as the rainiest year in these years, while 2016 had moderate weather conditions.

Aflatoxins contamination of maize in Serbia: the impact of weather conditions in 2015.

In recent years climate changes recorded in temperate regions of Europe have led to aflatoxin (AF) contamination of maize. Thus, the aim of this study was to investigate the influence of weather conditions on levels of aflatoxin B1 (AFB1), aflatoxin B2 (AFB2), aflatoxin G1 (AFG1) and aflatoxin G2 (AFG2) in 180 maize samples collected from the main maize-growing regions (Western Backa, North Banat, South Banat and Central Serbia) in Serbia after harvest in 2015. The concentrations of AFs were determined by a validated HPLC method with post-column derivatisation and fluorescence detection (HPLC-FLD). The presence of AFB1, AFB2, AFG1 and AFG2 was detected in 57.2%, 13.9%, 5.6% and 2.8% of maize samples in the concentration ranges of 1.3-88.8 µg kg-1, 0.60-2.8 µg kg-1, 1.8-28.5 µg kg-1 and 2.1-7.5 µg kg-1 respectively. The recorded smaller amount of precipitation and especially higher air temperatures during the summer of 2015 were favourable for AF production, which resulted in 32.2% and 21.1% of samples being unsuitable for human consumption, since AFB1 and the sum of AFs concentrations were above 5.0 and 10.0 µg kg-1 respectively. Furthermore, the findings in this study indicate that the microclimate conditions in the investigated regions had a great influence on the contamination frequency of maize with AFs. The highest percentage of samples unsuitable for human consumption, considering both AFB1 and total AFs content were 72.5% and 51.5% respectively from Central Serbia, whilst the lowest percentages of 15.6% and 6.2% respectively were found in Western Backa. These findings confirmed that maize should be continuously monitored in order to protect human and animal health from the harmful effects caused by AFs contamination.

Comparison of ELISA, HPLC-FLD and HPLC-MS/MS Methods for Determination of Aflatoxin M1 in Natural Contaminated Milk Samples.

Presence of aflatoxin M1 (AFM1) in milk should be continuously controlled in order to protect the population from risks associated with its proven toxicity and carcinogenicity. During recent years, there has been an increase in demand for development of sensitive, accurate, simple and fast method which is reliable for detection of AFM1 at low concentrations found in milk samples. For that purpose, enzyme linked immunosorbent asssay (ELISA), high performance liquid chromatography with fluorescence detector (HPLC-FLD) and high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) were optimized and validated in order to apply them for AFM1 analysis in naturally contaminated milk samples, and to assess the closeness of agreement between results of three different methods. The obtained validation parameters indicate that all three methods are suitable for determination of AFM1 in milk samples. The statistical analysis of variance between the methods and the obtained correlation coefficients indicate that there is a strong correlation between methods. All three methods are satisfactory in meeting the requirements for official control purposes. To the best of author's knowledge, this study represents the first report of an investigation and comparison of ELISA, HPLC-FLD and HPLC-MS/MS methods for determination of AFM1 in naturally contaminated milk samples.

Valorisation of raspberry and blueberry pomace through the formulation of value-added gluten-free cookies.

Successful valorisation of raspberry and blueberry pomace was achieved through their application, as dried and ground powders, in the formulation of value-added gluten-free cookies. Simplex-lattice mixture design approach was applied to obtain the product with the best sensory properties, nutritional profile and antioxidant activity. The highest desirability of 90.0 % was accomplished with the substitution of gluten-free flour mixture with 28.2 % of blueberry and 1.8 % of raspberry pomace. The model was verified. Optimized cookies had similar protein (3.72 %) and carbohydrate (66.7 %) contents as gluten-containing counterparts used for comparison, but significantly lower fat content (10.97 %). Daily portion of the optimized cookies meets: 5.00 % and 7.73 % of dietary reference intakes (DRIs) for linoleic acid, 23.6 % and 34.3 % of DRIs for α-linolenic acid and 10.3 % and 15.6 % of DRIs for dietary fibers, for male and female adults, respectively. The nutritional profile of the optimized formulation makes it comparable with added-value gluten-containing counterparts.

Antioxidant and Physicochemical Properties of Hydrogen Peroxide-Treated Sugar Beet Dietary Fibre.

The aim of the present work was to examine if hydrogen peroxide treatment of sugar beet fibre that aimed at improving its physicochemical properties would impair its antioxidant potential. Three different sugar beet fibres were obtained from sugar beet - non-treated fibre (NTF) from sugar beet cossettes extracted with sulphurous acid, treated fibre (TF) from NTF treated with hydrogen peroxide in alkaline solution and commercially available Fibrex(®) . The antioxidant activity of extractable and non-extractable fibre fractions in ethanol/water mixture (80:20, v/v) of three fibre samples was estimated. Non-extractable fractions obtained after alkaline treatment of investigated fibres were much higher in phenolic compounds and possessed higher antioxidant potential than extractable fractions. Ferulic acid was proven to be the dominant phenolic acid. Regarding both extractable and non-extractable fractions, Fibrex(®) had the highest antioxidant activity in chemical tests, while NTF was superior in comparison with TF. Based on the results of Caco-2 cells-based test, all non-extractable fractions possessed potential for reactive oxygen species inhibition. Regarding the extractable fractions, only the TF manifested this effect.Copyright © 2016 John Wiley & Sons, Ltd.

Sugar beet molasses as an ingredient to enhance the nutritional and functional properties of gluten-free cookies.

Sugar beet molasses is a raw material with high potential to be a functional ingredient in baked goods. This paper investigated the nutritional and functional properties of gluten-free cookies enriched with sugar beet molasses. At all enrichment levels and forms tested (liquid and dry), the addition of beet molasses improved the micronutrient pattern and antioxidative status of gluten-free cookies. The cookies prepared with molasses were significantly higher in potassium, magnesium, calcium, iron, betaine, total phenolics and DPPH radical scavenging abilities. Molasses contributed to wider spectra of phenolic compounds. The dominating phenolic compounds in the molasses-enriched cookies were catechin, ferulic, syringic and vanillic acid. Molasses also contributed to the presence of p-hydroxybenzoic acid in the cookies. Addition of molasses increased the content of hydroxymethyfurfural in the cookies, but not above values commonly reported for this product type. Molasses addition improved the overall acceptance of gluten-free cookies up to 30% enrichment level.

Characterization of byproducts originating from hemp oil processing.

Valorization of hemp seed meal, a byproduct of hemp oil processing, was performed by measuring the distribution of nutritional and antinutritional compounds in different hemp seed meal fractions. According to chemical composition, two cotyledon-containing fractions (>180 and <180 µm) were significantly richer in protein (p < 0.05) (41.2% ± 0.04% and 44.4% ± 0.02%, respectively), lipid (15.1% ± 0.02% and 18.6% ± 0.04%, respectively), and sugar content (4.96% ± 0.11% and 3.46% ± 0.08%, respectively) in comparison to the hull-containing fractions (>350 and >250 µm), which were significantly richer in crude fiber content (29.5% ± 0.04% and 21.3% ± 0.03%, respectively). The free radical scavenging capacity (IC50) of fraction extracts increased (p < 0.05) with increasing mean particle size (from 17.18 ± 0.59 to 5.29 ± 0.30 mg/mL). Cannabisin B and N-trans-caffeoyltyramine were the most abundant phenolic compounds in the hull fractions (from 267 ± 15.9 to 287 ± 23.1 mg/kg), while cotyledon fractions had higher content of catechin (from 313 ± 12.4 to 744 ± 22.2 mg/kg) and p-hydroxybenzoic acid (from 124 ± 6.47 to 129 ± 8.56 mg/kg (P < 0.05). Well-balanced ω-6 to ω-3 fatty acid ratio (3:1) was determined in all fractions. Antinutrients (trypsin inhibitors, phytic acid, glucosinolates, and condensed tannins) were mostly located in the cotyledon fractions. These findings indicate that the separation of hemp seed meal into different fractions could be used to concentrate valuable target compounds and consequently facilitate their recovery.

Multi-residue method for determination of selected neonicotinoid insecticides in honey using optimized dispersive liquid-liquid microextraction combined with liquid chromatography-tandem mass spectrometry.

The objective of this study was to develop analytical method based on optimized dispersive liquid-liquid microextraction (DLLME) as a pretreatment procedure combined with reversed phase liquid chromatographic separation on C18 column and isocratic elution for simultaneous MS/MS determination of selected neonicotinoid insecticides in honey. The LC-MS/MS parameters were optimized to unequivocally provide good chromatographic separation, low detection (LOD, 0.5-1.0 µg kg(-1)) and quantification (LOQ, 1.5-2.5 µg kg(-1)) limits for acetamiprid, clothianidin, thiamethoxam, imidacloprid, dinotefuran, thiacloprid and nitenpyram in honey samples. Using different types (chloroform, dichloromethane) and volumes of extraction (0.5-3.0 mL) and dispersive (acetonitrile; 0.0-1.0 mL) solvent and by mathematical modeling it was possible to establish the optimal sample preparation procedure. Matrix-matched calibration and blank honey sample spiked in the concentration range of LOQ-100.0 µg kg(-1) were used to compensate the matrix effect and to fulfill the requirements of SANCO/12495/2011 for the accuracy (R 74.3-113.9%) and precision (expressed in terms of repeatability (RSD 2.74-11.8%) and within-laboratory reproducibility (RSDs 6.64-16.2%)) of the proposed method. The rapid (retention times 1.5-9.9 min), sensitive and low solvent consumption procedure described in this work provides reliable, simultaneous, and quantitative method applicable for the routine laboratory analysis of seven neonicotinoid residues in real honey samples.