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The presence of mycotoxins and other toxic metabolites in hops (Humulus lupulus L.) was assessed for the first time. In total, 62 hop samples were sampled in craft breweries, and analyzed by a multi-toxin LS-MS/MS method. The study collected samples from craft breweries in all of the Croatian counties and statistically compared the results. Based on previous reports on Alternaria spp. and Fusarium spp. contamination of hops, the study confirmed the contamination of hops with these toxins. Alternaria toxins, particularly tenuazonic acid, were found in all tested samples, while Fusarium toxins, including deoxynivalenol, were present in 98% of samples. However, no Aspergillus or Penicillium metabolites were detected, indicating proper storage conditions. In addition to the Alternaria and Fusarium toxins, abscisic acid, a drought stress indicator in hops, was also detected, as well as several unspecific metabolites. The findings suggest the need for monitoring, risk assessment, and potential regulation of Alternaria and Fusarium toxins in hops to ensure the safety of hop usage in the brewing and pharmaceutical industries. Also, four local wild varieties were tested, with similar results to the commercial varieties for toxin contamination, but the statistically significant regional differences in toxin occurrence highlight the importance and need for targeted monitoring.

Viticulture has been an important economic sector for centuries. In recent decades, global wine production has fluctuated between 250 and almost 300 million hectoliters, and in 2022, the value of wine exports reached EUR 37.6 billion. Climate change and the associated higher temperatures could favor the occurrence of ochratoxin A (OTA) in wine. OTA is a mycotoxin produced by some species of the genera Aspergillus and Penicillium and has nephrotoxic, immunotoxic, teratogenic, hepatotoxic, and carcinogenic effects on animals and humans. The presence of this toxin in wine is related to the type of wine—red wines are more frequently contaminated with OTA—and the geographical location of the vineyard. In Europe, the lower the latitude, the greater the risk of OTA contamination in wine. However, climate change could increase the risk of OTA contamination in wine in other regions. Due to their toxic effects, the development of effective and environmentally friendly methods to prevent, decontaminate, and degrade OTA is essential. This review summarises the available research on biological aspects of OTA prevention, removal, and degradation.

The search for unexplored plant resources that would provide a good basis for the development of novel probiotic functional foods is rapidly increasing. In this context, the strawberry tree fruit (Arbutus unedo L.) is particularly interesting, as it is rich in numerous antioxidant bioactive compounds that have been shown to be beneficial to health, but have not yet found industrial applications. In this work, the probiotic characterization of lactic acid bacteria strain Lactiplantibacillus plantarum DB2, isolated from strawberries (Fragaria ananassa × Duch.), was performed. The tested strain proved to be safe to use, displaying no antibiotic resistance or hemolytic activity. Due to its proven probiotic potential during simulated gastrointestinal transit, its antimicrobial activity, and its coaggregation with pathogens, it was selected for fermentation of an aqueous Arbutus unedo L. extract, which was subsequently microencapsulated and freeze-dried to extend its shelf life and preserve its functional properties. The antioxidant activity of the ferment obtained was maintained (80%), while after microencapsulation and freeze-drying, about 50% and 20% of the antioxidant activity was retained, respectively. In conclusion, this study demonstrates for the first time the application of probiotics isolated from strawberries in the fermentation of strawberry tree extract and monitors the antioxidant activity during post-fermentation formulation, paving the way for a potential industrial application of this underutilized plant.

Climate change, especially in the form of temperature increase and sea acidification, poses a serious challenge to the sustainability of aquaculture and shellfish farming. In this context, lactic acid bacteria (LAB) of marine origin have attracted attention due to their ability to improve water quality, stimulate the growth and immunity of organisms, and reduce the impact of stress caused by environmental changes. Through a review of relevant research, this paper summarizes previous knowledge on this group of bacteria, their application as protective probiotic cultures in mollusks, and also highlights their potential in reducing the negative impacts of climate change during shellfish farming. Furthermore, opportunities for further research and implementation of LAB as a sustainable and effective solution for adapting mariculture to changing climate conditions were identified.

The aim of this paper was to examine the effect of different OTA concentrations on the parameters of oxidative stress (glutathione (GSH) and malondialdehyde (MDA) concentrations) and glucose utilization in ethanol production by wine yeasts. In addition to the above, artificial neural networks (ANN) were used to predict the effects of different OTA concentrations on the fermentation ability of yeasts and oxidative stress parameters. The obtained results indicate a negative influence of OTA (4 µg mL−1) on ethanol production after 12 h. For example, K. marxianus produced 1.320 mg mL−1 of ethanol, while in the control sample 1.603 µg mL−1 of ethanol was detected. However, after 24 h, OTA had no negative effect on ethanol production, since it was higher (7.490 and 3.845 mg mL−1) in comparison to control samples. Even low concentrations of OTA affect GSH concentrations, with the highest being detected after 12 and 24 h (up to 16.54 µM), while MDA concentrations are affected by higher OTA concentrations, with the highest being detected at 24 h (1.19 µM). The obtained results with the use of ANNs showed their potential for quantification purposes based on experimental data, while the results of ANN prediction models have shown to be useful for predictions of what outcomes different concentrations of OTA that were not part of experiment will have on the fermentation capacity and oxidative stress parameters of yeasts.

Pomegranate (Punica granatum L.) is well known for its high content of bioactives, including polyphenols, flavonoids, and tannins, which have been shown to exhibit a wide range of biological activities, such as antioxidant, antimicrobial, and anticancer effects. It is worth noting that the majority of these molecules are found in the peels, which are usually disposed of after processing, causing a significant amount of waste, amounting to more than 3.6 million t/y. This work investigates microwave-assisted extraction (MAE) in water for the recovery of antioxidants from pomegranate peels (PP), including the optimisation of temperature and extraction times. The total phenolic, anthocyanin, flavonoid, and tannin contents of the recovered extracts were determined, as well as their antioxidant activities, which were found to be 356.35 mgGAE/gExtr, 303.97 µgCy3G/gExtr, 37.28 mgQE/gExtr, 56.48 mgGAE/gExtr, and 5.72 mmolTE/gExtr, respectively (according to the adopted reference). All results were compared with those obtained using a conventional protocol. In addition, the potential for water recycling by means of downstream nanofiltration in optimised MAE was investigated, leading to overall water reuse of approx. 75%. Power consumption (20.92 W/mgGAE) and common green metrics, Reaction Mass Efficiency (RME), E-Factor, and the Process Mass Intensiti/efficiency (PMI, PME), were considered in evaluating the proposed PP valorisation strategy. Finally, the biological activities of the main products were assessed. The antimicrobial properties of the PP extracts against three Gram-positive and three Gram-negative bacteria and their antiproliferative activity towards human cancer cells were tested. S. aureus bacteria was the most susceptible to the PP extracts. All tested products displayed antiproliferative activity against HeLa cells when higher concentrations were tested, with D-PP/NF (obtained from dried PP and sequential nanofiltration) being the most effective. This result was also confirmed via clonogenic analysis, which generally indicated the possible anti-cancer activity of pomegranate peel extracts obtained using this green approach.

The objective of this research was to isolate and identify lactic acid bacteria living in the digestive tract of the queen scallop Aequipecten opercularis and further test it as a probiotic culture on the growth of the bivalve during one month in captivity. Classic microbiological techniques (API and MALDI TOF) were used and four different strains of Lactiplantibacillus and one Lactococcus were identified and further analyzed, namely: Lactiplantibacillus plantarum I, Lactiplantibacillus plantarum 1, Lactiplantibacillus plantarum 2, Lactococcus lactis, and Lactiplantibacillus brevis. The isolated cultures were further tested for the highest antimicrobial activity towards the most common marine pathogens and survival at different temperatures and pH levels. The strain Lactiplantibacillus plantarum I had the best results, with the highest antimicrobial activity (77–85% pathogen growth inhibition) and the best safety standards showing no antibiotic resistance, and no ability to synthesize biogenic amine and degrade red blood cells. Considering all the abovementioned characteristics, the strain Lpb. plantarum I was further tested on the growth and health status of the bivalve Aequipecten opercularis during a month of cultivation in captivity. The results showed that the incorporation of Lpb. plantarum I led to a decrease in the quantity of test microorganisms in the bivalve and an increase in both the growth rate weight and growth rate length of the queen scallop A. opercularis. Furthermore, the scallops fed with the algae culture + Lpb. plantarum I gained significantly higher meat yield (33.15 ± 2.63%) compared with the control scallops fed only with the algae culture (29.66 ± 2.87%). To conclude, the results from this research indicate that including Lpb. plantarum I as a dietary supplement could enhance growth performance and serve as a feasible approach to reduce pathogen levels while cultivating A. opercularis in captivity.

3D printing technology (3DP) as additive manufacturing is an innovative design technology that can meet the individual nutritional and sensory needs of consumers. Therefore, the aim of this work was to apply 3DP in the production of a strawberry-based functional product with the addition of two hydrocolloids (corn and wheat starch) in three proportions (10, 15 and 20%) and to investigate the influence of 3DP process parameters on physico-chemical and textural properties, as well as the bioactive and antioxidant potential and microbiological stability, with(out) the addition of natural antimicrobial agents. Starch type had a significant effect on all tested bioactive compounds, as well as on starch content, except for total phenolic and hydroxycinnamic acid contents. Considering the content of bioactive compounds and antioxidant capacity, program 2 proved to be more suitable than program 1. All samples exhibited good textural properties, a high degree of stability and minimal geometric deviations. Regarding microbiological safety, no pathogenic bacteria were found in the 3DP samples during storage. The 3DP sample with added citral at a concentration of 75 mg L−1 showed the best microbiological quality. Ultimately, 3DP can be successfully used for the production of new strawberry-based functional products.

Research background. Cheese in a sack is a traditional cheese produced in Croatia. Types of cheese with similar production technology are made in other countries but chemical and microbiological composition varies between regions. Traditionally, cheese in a sack is produced without the addition of starter cultures. Addition of beneficial probiotic cultures to numerous dairy products has documented advantages. Effects that the addition of probiotic bacteria to traditional cheese have on aroma compounds and sensory properties have not been fully investigated. The aim of this study is to determine the sensory properties and differences in the aromatic profiles between cheese samples ripened in a lambskin sack, produced traditionally without the addition of any starter culture, or with the addition of probiotic bacteria. Experimental approach. In this study, cheese in a sack was produced with the addition of probiotic cultures Lactobacillus plantarum B and L. lactis ssp. lactis S1. During ripening volatile aroma compounds were analysed with a solid-phase microextraction gas chromatography-mass spectrometry. Sensory properties were evaluated by trained tasters who are familiar with the traditional taste of the cheese from a sack. The results of aroma composition and taste scores were then compared using factorial and principal component analyses. Results and conclusions. Chromatography showed differences in the composition of aroma compounds and the sensory properties between the cheese produced with Lactobacillus starter cultures and the control cheese, traditionally produced without a starter culture. The addition of probiotic cultures L. plantarum B and L. lactis ssp. lactis S1 resulted in products with better sensory properties and chemical profile of volatile aromatic compounds. Novelty and scientific contribution. This study investigates the usage of naturally present probiotic cultures as starter cultures in cheese in a sack production. Their effects on aroma profiles and sensory characteristics have been compared for the first time using factorial and principal component analyses.

During ripening, the surface of dry traditional meat products (TMPs) becomes overgrown by fungi of the Penicillium spp., Aspergillus spp. and Eurotium spp. whose spores mostly come from the environment in which the ripening chambers are placed. Certain fungi species is often responsible for the occurrence of toxic compounds termed the mycotoxins, among which of the outermost importance in connection with meat products are aflatoxin B1 (AFB1) and ochratoxin A (OTA). Besides, some other mycotoxins such as citrinin (CIT), cyclopiazonic acid (CPA) and sterigmatocystin (STC) can also be present, but their impact on the quality and safety of meat products, and therefore also on human health, has still not been fully clarified. As control and prevention of toxicogenic fungi growth are key factors to the prevention of mycotoxin presence in dry-cured TMPs, levels of mycotoxin contamination, mycotoxin-producing mould species and factors of relevance for mycotoxin production, such as climate, should be determined.

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.

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.

Lactobacillus plantarum O1 was isolated from the gut of sea bream (Sparus aurata) and identified with the API biochemical test and MALDI TOF MS. This strain was further characterised according to the selection criteria for lactic acid bacteria as starter cultures for aquatic food production. L. plantarum O1 showed good antimicrobial activity against pathogenic test microorganisms. Further investigation confirmed it as the producer of the bacteriocin plantaricin. This strain also showed good growth at a wide range of temperatures (from 4 to 45 °C) and a wide range of pH (2–12), even in the presence of 3.5 % NaCl. Its viability was also good after lyophilisation and in simulated gastric and small intestinal juice. The strain is a promising probiotic, and our further research will focus on its application in the biopreservation of fresh fish and shellfish.

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.

As in the traditional production of cheese in lambskin sacks raw cow’s or sheep’s milk is mostly used, the purpose of this study is to see how the production affects the microbiological quality of the cheese. To do that, we tested 39 samples of raw cow’s and sheep’s milk, curd, ripened cheese (15, 30 and 45 days) and lambskin sacks for native microbial population. Two-thirds of the milk, curd and cheese samples had higher counts of staphylococci and enterobacteria than permitted by regulations. Not a single sample had Salmonella and Listeria monocytogenes, but we did find Escherichia coli in sheep’s milk and cheese, and yeast and mould in both types of milk and cheese. Staphylococcus xylosus prevailed in lambskin sacks. Despite the high incidence of S. aureus, even in the final product, staphylococcal enterotoxin was detected in only two sheep’s cheese samples. Among the lactic acid bacteria, Lactococcus lactis and Lactobacillus paracasei prevailed in cow’s cheese, whereas Leuconostoc mesenteroides and Lactobacillus plantarum prevailed in sheep’s cheese. In the lambskin sacks Leuconostoc mesenteroides and Lactobacillus plantarum were predominant. Our findings give an important insight into the fermentation and microbial ecology of the cheese in lambskin sacks.

Cheese is a highly valued dairy product worldwide, with a special focus on traditional cheeses, not only for their basic nutritive purposes but also as a part of the culture and characteristics of a certain country. Owing to the geographical location as well as the climate and vegetation diversity, in certain regions of Croatia the production of various traditional cheeses using spices was developed. Spices are either added to the cheese curd which is then formed, or cheese is wrapped into plant leaves. Sometimes spices are also applied onto the surface of the cheese, and only in rare cases spices are added into the curd. Spices added to cheese improve sensory characteristics, increase the stability and shelf life but also increase the nutritional value. The chemical composition of spices is very complex and every spice has a specific and dominant ingredient that contributes to the flavour of the product and/or its antimicrobial and antioxidant activity. This paper provides an overview of spices and aromatic herbs as natural preservatives that are used in the production of traditional cheeses.

Limbal Stem Cell Deficiency (LSCD) is a very serious and painful disease that often results in impaired vision. Cultivation of limbal stem cells for clinical application is usually performed on carriers such as amniotic membrane or surgical fibrin gel. Transplantation of these grafts is associated with the risk of local postoperative infection that can destroy the graft and devoid therapeutic benefit. For this reason, electrospun scaffolds are good alternatives, as proven to mimic the natural cells surroundings, while their fabrication technique is versatile with regard to polymer functionalization and scaffolds architecture. This study considers the development of poly(ε-caprolactone) (PCL) immune-compatible and biodegradable electrospun scaffolds, comprising cefuroxime (CF) or titanium dioxide (TiO2) active components, that provide both bactericidal activity against eye infections and support of limbal stem cells growth in vitro. The PCL/CF scaffolds were prepared by blend electrospinning, while functionalization with the TiO2 particles was performed by ultrasonic post-processing treatment. The fabricated scaffolds were evaluated in regard to their physical structure, wetting ability, static and dynamic mechanical behaviour, antimicrobial efficiency and drug release, through scanning electron microscopy, water contact angle measurement, tensile testing and dynamic mechanical analysis, antimicrobial tests and UV-Vis spectroscopy, respectively. Human limbal stem cells, isolated from surgical remains of human cadaveric cornea, were cultured on the PCL/CF and PCL/TiO2 scaffolds and further identified through immunocytochemistry in terms of cell type thus were stained against p63 marker for limbal stem cells, a nuclear transcription factor and cytokeratin 3 (CK3), a corneal epithelial differentiation marker. The electrospun PCL/CF and PCL/TiO2 successfully supported the adhesion, proliferation and differentiation of the cultivated limbal cells and provided the antimicrobial effect against Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans.

The study focuses on developing novel cott age cheese containing spices with acceptable sensory properties, increased biological value and extended shelf life. Thirty types of cheese with added fresh or dried parsley, dill, pepper, garlic and rosemary were produced. Characterisation of phenolic compounds, antioxidant capacity and antibacterial activity of spices and cheese samples were evaluated. The cheese containing fresh pepper and fresh and dried herbs showed excellent sensory properties, with the best results obtained with fresh sweet red pepper. Dry rosemary had the highest antioxidant and antibacterial activity due to high mass fractions of caffeic and rosmarinic acids as well as high mass fractions of flavones and phenolic diterpenes. The plant extracts examined in vitro and in situ effectively reduce numbers of foodborne pathogens like Salmonella typhimurium, Escherichia coli, Staphylococcus aureus and Listeria monocytogenes, and therefore have potential as natural preservatives and antioxidants.

The aim of this research is to use isolated and characterized autochthonous functional starter cultures from traditional Croatian dry sausages and to evaluate their capacity for industrial production of five sausages (Čajna sausage, Zimska sausage, Bečka sausage, Srijemska sausage and Slavonski kulen). These defined autochthonous functional starter cultures (combination of Lactobacillus and Staphylococcus strains) were used to produce five different industrial sausages which were compared by a panel. The viability of introduced autochthonous Lactobacillus and Staphylococcus strains and their effect on the final product characteristics, namely microbiological, physicochemical and sensory properties were monitored. The obtained results indicate that autochthonous starter cultures survived industrial production of sausages and can be used for production of sausages under controlled conditions. Autochthonous starter cultures obtained better results in the organoleptic evaluation, microbial safety and prolonged shelf-life in comparison with commercial starter cultures.

The aim of this study is to determine the effect of high intensity ultrasound (amplitude, temperature and treatment time) on the inactivation of food spoilage bacteria Escherichia coli 3014, Staphylococcus aureus 3048, Salmonella sp. 3064, Listeria monocytogenes ATCC 23074 and Bacillus cereus 30. The model suspensions of bacteria were treated with 12.7-mm ultrasonic probe operated at 600 W nominal power (ultrasonic treatment implemented at 20 kHz) and at amplitudes of 60, 90 and 120 µm. Also, treatment time of 3, 6 and 9 min and temperature of 20, 40 and 60 °C were used. The results were statistically processed with STATGRAPHICS Centurion computer program and response surface methodology. All three parameters studied seem to substantially affect the inactivation of bacteria in pure culture. The results also indicate increased inactivation of microorganisms under longer period of treatments, particularly in combination with higher temperature and/or amplitude. After ultrasonic treatment at 60 °C, 9 min and 120 μm, the viability of cells was not confirmed for Escherichia coli 3014, Staphylococcus aureus 3048, Salmonella sp. 3064 and Listeria monocytogenes ATCC 23074. Under the mentioned conditions the highest inactivation (3.48 log CFU/mL) of Bacillus cereus 30 was obtained.

The purpose of this research was to determine whether the cream cheese and cream that are produced in the traditional manner at home and are free to sale on Zagreb markets, meet microbiological requirements for foodstuffs (OG 46/94, 20/01, 40/01). Particular attention is given to research of bacteria Listeria monocytogenes presence in these foods, because of its exceptional hazards to human health. It was found that a majority of 64 (53 %) from a total of 120 studied dairy products samples were contaminated with microbial pathogens, of which 16 % are waste in the cream cheese, and 37 % in cream samples. 39 samples of cheese and 50 samples of cream did not fulfil the conditions prescribed by the Croatian Guidelines, primarily due to the contamination with yeasts and moulds. In 10 cheese and cream samples where L. monocytogenes is proven by classical microbiological methods, PCR method confirmed L. monocytogenes in only one cream sample.

The objective of this paper is to emphasize, branding as one of the possible ways to take concrete steps towards reaffirming the local values and traditional knowledge of rural populations in Serbia. Based on information gathered in the field, as well as from existing literature sources, the main goal of this study is to promote products from the Visok microregion. Due to very alarming demographic data, this may be the last chance for revitalizing this region. In this study, we highlighted the following products: Pirot kilim (Pirot rug), Pirot/Stara Planina lamb, Pirot Kachkaval cheese, Stara Planina honey, and Stara Planina potato. Besides agriculture, micr oregion Visok also has exceptional tourist potential. The conclusion of this paper emphasizes that the only viable path to revitalization is through planned investment. Such investment would facilitate the development of local infrastructure and help restore the population, thus enabling the creation of sustainable development.