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Mycotoxins are secondary metabolites of moulds that represent a significant food safety issue and pose a risk to health and wellbeing of humans and animals, having a negative impact on economies all over the world. They can cause intoxication in animals and affect the less productive performance and nutritional value of animal feed. Also, mycotoxins may contaminate different types of food through directly and indirectly way and cause negative health effects in humans. To estimate the potential of their harmful effect it is necessary to know the terms of mycotoxins occurrence and frequency, toxicity and biotransformation in different animal species and humans, possibility of contamination prevention, and their inactivation or reduction in the case when the incidence is inevitable. The relatively small number of comparable data on the incidence of mycotoxins in feed from European countries show the necessity of systematic control and the application of validated analytical methods in their determination. This paper gives the overview of incidence of grains and feed contamination with most important and common mycotoxins and the evidenced toxic effects in animals.

Diospyros kaki L. (Japanese persimmon) has been introduced from Asian countries to Herzegovina, where it is mostly consumed fresh. This study investigates the differences in nutritional, microbiological, and sensory values of persimmons consumed in one of the Herzegovinian regions, following drying processes at 70 °C/12 h, 85 °C/10.5 h, and 100 °C/5.5 h, together with their effects on fruit texture, mass, and composition. The notable increase in sugar content emphasises the potential of dried Japanese persimmon as a nutritious snack. No significant increase in microorganism representation was observed across all samples. Persimmons dried using a dehydrator exhibited the most preserved nutritional properties, and were rated highest in sensorial evaluations. However, due to its high water content and browning, the sample dried at 100 °C in an autoclave was excluded from sensory testing. [ABSTRACT FROM AUTHOR]

The aim of this study was to establish the differences in fungi and mycotoxin contamination and mycobiota biodiversity between wheat and maize after harvesting. Determination of fungal contamination of wheat and maize is crucial, given that these grains are extensively used in food and feed production. Some moulds can produce toxic secondary metabolites, such as mycotoxins, which can cause serious health problems in humans and animals. Due to the metabolic activities of moulds, undesirable changes in food and feed colour, taste, and smell can occur. In this study, samples of wheat and maize, harvested during 2021 in the Northern Croatia, were analysed for mycological and mycotoxin contamination before storage. Also, potentially mycotoxigenic mould genera were identified at the species level in order to uncover the presence of toxigenic species. The presence of the following mycotoxins was examined: zearalenone, deoxynivalenol, ochratoxin, aflatoxin B1, T-2, and HT-2 toxin. The results indicate that both wheat and maize samples were contaminated with yeasts, with maize exhibiting a higher level of contamination as compared to wheat (1.5 × 104 to 1.8 × 106 vs 1.7 × 103 to 9.0 × 105). In wheat, the predominant mycotoxigenic mould species was Alternaria, as opposed to maize in which the Penicillium genus was the most represented. Mycotoxin analysis revealed that both maize and wheat were contaminated with Fusarium mycotoxins. Aflatoxin B1 and ochratoxin were not determined in any of the tested samples. The results point out that grains carry a different risk of contamination with Fusarium mycotoxins and that maize is more susceptible to fungus spoilage.

The aim of this study was to investigate nutritive and sensorial quality of Istrian and Slavonian pork-meat dry-fermented sausages. Sensorial analysis resulted in statistically significant differences (p<0.05) between these products in 11 sensorial parameters analysed. Significantly higher protein content was determined in Istrian in comparison with Slavonian dry-fermented sausage, although the latter content significantly varied across the producing households. No significant differences in particular fatty acid esters and saturated, monounsaturated and polyunsaturated fatty acids were obtained. Although some of the nutritional quality indices failed to meet health recommendations, the obtained values are consistent with data published for pork meat products. [ABSTRACT FROM AUTHOR]

Nutritive composition, fatty acid profile and health-related lipid indices of natural-born European flat oyster (Ostrea edulis), variegated scallop (Chlamys varia) and smooth scallop (Flexopecten glaber) in 108 samples originating from the Adriatic sea, recovered on a monthly basis were investigated. Out of three shellfish species, the lowest share of saturated fatty acids, the most favourable ratio of polyunsaturated over saturated fatty acids, the most favourable atherogenic and thrombogenic index, and the most favourable ratio of hypocholesterolaemic over hypercholesterolaemic fatty acids were seen in oysters, sampled during springtime. No statistically significant inter-seasonal differences between basic chemical parameters and fat quality indices were established. [ABSTRACT FROM AUTHOR]

Metabolic transformation of zearalenone (ZEA), a mycotoxin which can contaminate both food and feed, results in the formation of five metabolites, one of them being zeranol (α-ZAL), which can be abused in farm animals as a growth promoter. To the best of our knowledge, there is no analytical method that can distinguish whether α-ZAL is present in an animal urine sample as a result of ZEA biotransformation or as a result of anabolic abuse. This study aimed at monitoring resorcylic acid lactones (RALs) concentration in urine of farm animals over several years. Six hundred and three cattle and pig urine samples were collected on farms in different Croatian regions and analyzed for RAL presence. Based on the testing results, all RAL-positive samples were considered to be consequential to feed contamination. The difference in primary ZEA metabolites' ratio (α-zearalenol/β-zearalenol) was observed between cattle (0.03–0.41) and pig (2.05–17.39) urine samples. If the animals are treated with α-ZAL and fed on ZEA-contaminated feed, α-ZAL and taleranol found in their organisms could come from two sources, so that the reliability of the statistical model might be questionable. Based on these findings, there exists the need for improving the approach to the distinction between α-ZAL abuse and ZEA feed contamination. [ABSTRACT FROM AUTHOR]

We investigated the nutritional quality of commercially important farmed fish species: sea bream (Sparus aurata), sea bass (Dicentrarchus labrax), dentex (Dentex dentex), and turbot (Scophthalmus maximus). The omega-3 fatty acid content of dentex was twice as high as that of any other fish species, and its eicosapentaenoic (EPA) and docosahexaenoic (DHA) acid contents were 2-4 times higher. The recommended n-3/n-6 ratio was present in all the fish species, but the recommended polyunsaturated/saturated fatty acid ratio was not present in the turbot. All the fish species, except turbot, met the recommended atherogenic index, thrombogenic index, and ratio of hypocholesterolaemic to hypercholesterolaemic fatty acids, whereas the highest flesh lipid quality value was observed in the dentex. [ABSTRACT FROM AUTHOR]

The liquid chromatography-electrospray ionization tandem mass spectrometry method was developed for determination of clenbuterol residues in liver tissue as a regulatory matrix for the control of clenbuterol abuse as an anabolic. To investigate the level of clenbuterol residues during the withdrawal period, male food-producing pigs were exposed to subchronic repeat oral administration of a clenbuterol growth-promoting dose for 28 days. The analytical procedure showed acceptable validation results for all liver spiked samples analyzed and proved to be useful as a quantification and confirmation method in supporting regulatory enforcement programs of clenbuterol misuse monitoring. The highest level of clenbuterol in the liver of treated animals was recorded on day 0 of treatment cessation (21.58±14.29 ng/g), followed by 6.59±3.11 ng/g on day 3, 0.83±0.27 ng/g on day 7, and 0.44±0.08 ng/g on day 14 of withdrawal. At the end of the study period (day 35), the concentration of clenbuterol was below the method limit of detection (

Changes in physicochemical, colour, textural, microbiological and sensory attributes occurring during the processing of Horse Salami and established on manufacturing days 0, 7, 14, 21, 28, 42, 60, 90 were studied. Significant changes (P<0.05) in physicochemical parameters attributable to moisture loss, as well as changes in colour and textural properties were observed during the fermentation and ripening stage. Proteolysis and lipolysis, coming as a result of endogenous enzymatic activity and high lactic acid bacteria and staphylococci counts, contributed to specific organoleptic properties of the final product. Sensorial profiling showed a significant (P<0.05) acid taste, lactic acid odour and flavour intensity, and low fat/lean ratio and smokiness and saltiness values. Final Horse Salami products were microbiologically safe, the dominant microbial population thereby being Lactobacillus plantarum, Lactococcus lactis ssp. lactis, Enterococcus faecium and Staphylococcus xylosus. [ABSTRACT FROM AUTHOR]

The study aims to evaluate the presence of 15 polycyclic aromatic hydrocarbons (PAHs) in fishery products, shellfish, meat products and spices (n = 140). Benzo[a]pyrene (BaP) was detected in the mean concentration of 0.11 µg/kg in mussels to 4.85 µg/kg in spices. However, none of the samples exceeded the maximal BaP and ∑PAH4 limit, set out under the European legislation, although high values determined in some food, especially dried herbs and spices, pointed towards a heavy contamination and the necessity for systematic controls. The study showed that processed food samples contained significantly higher (p<0.05) PAH levels in comparison to food coming from environmental sources. [ABSTRACT FROM AUTHOR]

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.

During the last decade, scientists have given increasingly frequent warnings about global warming, linking it to mycotoxin-producing moulds in various geographical regions across the world. In the future, more pronounced climate change could alter host resilience and host-pathogen interaction and have a significant impact on the development of toxicogenic moulds and the production of their secondary metabolites, known as mycotoxins. The current climate attracts attention and calls for novel diagnostic tools and notions about the biological features of agricultural cultivars and toxicogenic moulds. Since European climate environments offer steadily rising opportunities for Aspergillus flavus growth, an increased risk of cereal contamination with highly toxic aflatoxins shall be witnessed in the future. On top of that, the profile (representation) of certain mycotoxigenic Fusarium species is changing ever more substantially, while the rise in frequency of Fusarium graminearum contamination, as a species which is able to produce several toxic mycotoxins, seen in northern and central Europe, is becoming a major concern. In the following paper, a high-quality approach to a preventative strategy is tailored to put a stop to the toxicogenic mould- and mycotoxin-induced contamination of foods and feeds in the foreseeable future.

The aim of this study was to investigate the microbial and physico-chemical characteristics of cold smoked sea bass (CSSB), a novel italian fish product. The microbiological analyses showed the presence of bacterial contamination from the raw material, the environment, and the production process. The microbial spoilage population was dominated by lactic acid bacteria (LAB) associated with Gram-negative fermenting bacteria, including Photobacterium phosphoreum and psychrotrophic Enterobacteriaceae. Brochotrix thermospacta and Aeromonas spp. were also present; in contrast, mould and yeast were not detected (<2 CFU/g). High levels (6-7 log CFU/g) of LAB and total bacteria count (TBC) were observed from day 45 of storage; however, their presence does not seem to have influenced the total volatile basic nitrogen (TVB-N), which always remained below 35 mg N/100 g. Consequently, the product is acceptable until day 60 of storage, considering that the malonaldehyde index (TBARS) was lower than 6.5 nmol/g. Pathogenic bacteria such as Salmonella spp. and Listeria monocytogenes were not detected. Currently, there is a growing demand for seafood due to its high quality and nutritional value. Cold smoked sea bass offers a source of macro- and micronutrients essential for the proper functioning of the human body. It is also rich in protein and omega-3 fatty acids. The WHO and FAO evaluated the benefits and risks and concluded that there is convincing evidence of health benefits from fish consumption, such as a reduction in the risk of heart failure and improved neurodevelopment in infants and young children when fish is consumed by the mother before and during pregnancy. The CSSB analysed in this study demonstrated to have health benefits due to long-chain omega-3 PUFAs and other nutrients, such as proteins, minerals, and vitamin D, which are sometimes difficult to obtain from other sources. The results show that CSSB has a high nutritional value and excellent microbial quality.

Ongoing climate change may affect the susceptibility of plants to attacks by pathogenic, mostly mycotoxigenic fungi with a consequent increase in the presence of mycotoxins. Fusarium fungi represent one of the most important producers of mycotoxins, and are also important pathogens of agricultural crops. Therefore, the main aim of the study was to estimate the impact of weather parameters on the natural occurrence of Fusarium mycotoxins, such as deoxynivalenol (DON), fumonisins B 1 and B 2 (FUMs), zearalenone (ZEN), T-2, and HT-2 toxins (T-2/HT-2) in maize samples harvested from two neighboring countries, Serbia and Croatia, during a four-year production period (2018-2021). The frequency and contamination level of examined Fusarium mycotoxins varied by maize year of production and could be linked to weather conditions per investigated country. Among them, FUMs were found to be the most common contaminants (84-100%) of maize in both Serbia and Croatia. Additionally, a critical assessment of Fusarium mycotoxins occurrence in the last 10 years (2012-2021), for both Serbia and Croatia, was done. Results pointed out the highest contamination of maize from 2014, especially with DON and ZEN, in connection to extreme levels of precipitation observed in both Serbia and Croatia, whereas FUMs occurred with high prevalence from each of the ten investigated years.

Aflatoxins (AFs) represent the most important mycotoxin group, whose presence in food and feed poses significant global health and economic issues. The occurrence of AFs in maize is a burning problem worldwide, mainly attributed to droughts. In recent years, Serbia and Croatia faced climate changes followed by a warming trend. Therefore, the main aim of this study was to estimate the influence of weather on AFs occurrence in maize from Serbia and Croatia in the 2018-2021 period. The results indicate that hot and dry weather witnessed in the year 2021 resulted in the highest prevalence of AFs in maize samples in both Serbia (84%) and Croatia (40%). In maize harvested in 2018-2020, AFs occurred in less than, or around, 10% of Serbian and 20% of Croatian samples. In order to conduct a comprehensive study on the implications of climate change for the occurrence of AFs in maize grown in these two countries, the results of available studies performed in the last thirteen years were searched for and discussed.

The influence of semi-hard (C1), hard (C2), and soft whey cheese (C3) immersed in extra virgin olive oil (EVOO) on its oxidative and hydrolytic parameters, fatty acids, and phenolic composition during two months of simultaneous storage was investigated. Accelerated hydrolytic and oxidative degradation was noted in EVOO stored with the immersed cheese compared to control oil. Oxidation indicator (K 232 ), myristic (C 14:0), and trans -oleic fatty acid (C18:1t) exceeded the prescribed limit for the EVOO category in oils stored with immersed C1 and C2, which indicated that standard analytical parameters are ineffective as tools to examine the declared quality and authenticity of such topping oils. The noted changes in fatty acid profile were primarily prescribed to the migration of fats. C1 and C2 influenced a comparable reduction in EVOO total identified phenolic content (-92.1% and -93.5%, respectively), despite having a different content of total proteins and moisture, whereas C3 influenced a slightly lower reduction (-85.0%). Besides the protein profile, other cheese compounds (e.g., moisture, carbohydrates) have been shown to have a considerable role in the development of the EVOO phenolic profile. Finally, compositional changes in EVOO used as a medium for cheese preservation are under significant influence of the cheese's chemical composition.

Today's increased demand and consumption of fish would be impossible to ensure without aquaculture. Farmed fish, however, is often considered inferior among consumers in comparison to its wild counterparts. The aim of this study was to profile Croatian fishery consumers based on their intention to consume farmed fish. The participants in this study were a nationally representative sample of people responsible for food purchasing within the household (n = 977), whose responses were collected by the CAWI (computer-aided web interviewing) method. Four clusters were identified and described: farmed fish enthusiasts (21.1%), farmed fish supporters (17.4%), indifferents (44.7%), and farmed fish sceptics (16.8%). Results showed that consumer segments differed significantly with respect to age, income, employment status, living region, and physical activity. Furthermore, intention to consume farmed fish is related to fish consumption in general (those with higher intention are more frequent fish consumers). Interestingly, prejudices against farmed fish are present in all clusters; however, these prejudices are more pronounced among those with the weakest intention to consume farmed fish. Differences between clusters were observed also in respect to product information and preferences, knowledge about fish, places of usual purchase, and source of information about fishery products. The obtained results could be used in designing marketing strategies to promote farmed fish consumption.

This study investigated the occurrence of 5 unregulated mycotoxins in a total of 250 traditional dry-cured meat products sampled in 2020 and 2021 in five Croatian regions (eastern, northern, central, western, and southern). Aflatoxin B 1 (AFB 1 ), ochratoxin A (OTA), sterigmatocystin (STC), citrinin (CIT), and cyclopiazonic acid (CPA) concentrations were related to the geographical region of the product's origin and to local weather. The results revealed the contamination of 27% of samples, namely, STC in 4% of samples in concentrations of up to 3.93 µg/kg, OTA in 10% of samples in concentrations of up to 4.81 µg/kg, and CPA in 13% of samples in concentrations of up to 335.5 µg/kg. No AFB 1 or CIT contamination was seen. Although no statistically significant differences in concentrations of individual mycotoxins across the production regions were found, differences in mycotoxin occurrence were revealed. The eastern and western regions, with moderate climate, delivered the largest number of contaminated samples, while the southern region, often compared with subtropics, delivered the smallest, so that the determined mycotoxins were probably mainly produced by the Penicillium rather than the Aspergillus species. Due to the interaction of various factors that may affect mycotoxin biosynthesis during production, the detected concentrations cannot be related solely to the weather.

Background: Increasing demand for high-value fish species and pressure on forage fish is challenging aquaculture to ensure sustainable growth by replacing protein sources in aquafeeds with plant and terrestrial animal proteins, without compromising the economic value and quality of the final fish product. In the present study, the effects of a plant protein-based diet (CV), two plant-based diets in which graded amounts of plan protein mixtures were replaced with Hermetia illucens meal alone (VH10) or in combination with poultry by-product meal (PBM) (VH10P30), a fishmeal (FM) diet (CF) and an FM diet supplemented with H. illucens (FH10) on growth performance, gut health and homeostasis of farmed subadult European seabass were tested and compared., Results: Fish fed the VH10 and VH10P30 diets showed the highest specific growth rates and lowest feed conversion ratios among the tested groups. Expectedly, the best preservation of PI morphology was observed in fish fed the CF or FH10 diets, while fish fed the CV diet exhibited significant degenerative changes in the proximal and distal intestines. However, PBM supplementation mitigated these effects and significantly improved all gut morphometric parameters in the VH10P30 group. Partial substitution of the plant mixture with insect meal alone or PBM also induced most BBM genes and activated BBM enzymes, suggesting a beneficial effect on intestinal digestive/absorption functions. Regarding intestinal microbiota, fish fed diets containing H. illucens meal (FH10, VH10, VH10P30) had the highest richness of bacterial communities and abundance of beneficial genera such as Lactobacillus and Bacillus. On the other hand, fish fed CV had the highest microbial diversity but lost a significant component of fish intestinal microbiota, the phylum Bacteroidetes. Finally, skin pigmentation most similar to that of farmed or even wild seabass was also observed in the fish groups fed CF, FH10 or VH10P30., Conclusion: Plant-based diets supplemented with PBM and H. illucens pupae meal have great potential as alternative diets for European seabass, without affecting growth performance, gut homeostasis, or overall fitness. This also highlights the importance of animal proteins in diets of European seabass, as the addition of a small amount of these alternative animal protein sources significantly improved all measured parameters., (© 2022. The Author(s).)

Due to its numerous health benefits, fish consumption should be strongly encouraged. Fish consumption, however, is a complex phenomenon influenced by various factors. The aim of this research is to examine the influence of knowledge, product information, and satisfaction with product attributes on fish consumption in a nationally representative sample of people responsible for food purchasing within households in Croatia (n = 977) and Italy (n = 967). Fish consumption was well predicted (R2 = 15%) by the proposed structural model, using the partial least squares structural equation modelling method (PLS-SEM). The obtained results confirm that subjective knowledge (β = 0.277, p < 0.001) and satisfaction with product attributes (β = 0.197, p < 0.001) are predictors of fish consumption. Subjective knowledge was influenced by product information (β = 0.161, p < 0.001), as well as by satisfaction with product attributes (β = 0.282, p < 0.001), while objective knowledge had an influence on product information (β = 0.194, p < 0.001). Although satisfaction with product attributes was the strongest predictor of subjective knowledge in both countries (βCRO = 0.244, βIT = 0.398), it had a greater effect among Italians (p = 0.001), while the impact of product information (βCRO = 0.210, βIT = 0.086) was more pronounced among Croatians (p = 0.010). Since the mediating role of subjective knowledge in all models was confirmed, action focused on enhancing subjective knowledge should be taken to increase fish consumption.

As filter feeders, bivalves and ascidians can accumulate contaminants present in the environment and pass them on to higher food chain levels as vectors. The consumption of bivalves contaminated with the potent neurotoxin domoic acid (DA) can cause amnesic shellfish poisoning in humans. The aim of this study was to determine seasonal differences in occurrence and accumulation of this phycotoxin in European oysters ( Ostrea edulis Linnaeus, 1758) ( n = 46), Queen scallops ( Aequipecten opercularis Linnaeus, 1758) ( n = 53), and edible ascidians of the Microcosmus spp. ( n = 107), originating from the same harvesting area in the Northern Adriatic Sea. The quantification was performed using ultra-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) preceded by derivatization with dansyl chloride. DA was found in very low concentrations throughout the year, with a maximum value of 810 μg/kg in Queen scallops. This study reveals differences in the occurrence and accumulation of DA between Queen scallops and the other two investigated species (oysters and ascidians) and the highest concentrations during the colder part of the year. Even though DA was detected in all of them, Queen scallops showed higher DA accumulation compared to the other two ( p < 0.001), hence representing a sentinel species suitable for the monitoring of DA level in seafood.

The aim of this study was to identify and compare surface mycobiota of traditional and industrial Croatian dry-fermented sausage Kulen , especially toxicogenic species, and to detect contamination with mycotoxins recognized as the most important for meat products. Identification of mould species was performed by sequence analysis of beta- tubulin and calmodulin gene, while the determination of mycotoxins aflatoxin B 1 (AFB 1 ), ochratoxin A (OTA), and cyclopiazonic acid (CPA) was carried out using the LC-MS/MS (liquid chromatography-tandem mass spectrometry) method. The results showed a significantly higher number of mould isolates and greater species (including of those mycotoxigenic) diversity in traditional Kulen samples in comparison with the industrial ones. P. commune , as a potential CPA-producer, was the most represented in traditional Kulen (19.0%), followed by P. solitum (16.6%), which was the most represented in industrial Kulen samples (23.8%). The results also showed that 69% of the traditional sausage samples were contaminated with either CPA or OTA in concentrations of up to 13.35 µg/kg and 6.95 µg/kg, respectively, while in the industrial samples only OTA was detected (in a single sample in the concentration of 0.42 µg/kg). Mycotoxin AFB 1 and its producers were not detected in any of the analysed samples (

Background: The objective of this study was to evaluate the efficacy of modified clinoptilolite (Minazel Plus ® , MZ) as a mycotoxin adsorbent for preventing the negative the effects of ochratoxin A (OTA) on performance, pathohistological changes, and OTA residue in the eggs of laying hens., Methods: Forty eight ( n = 48) laying hens (27 weeks old) were equally divided into six groups and depending on the type of addition were allocated to the following experimental treatments for 7 weeks: E-I group-1 mg/kg OTA; E-II group 0.25 mg/kg OTA; E-III group 1 mg/kg OTA + 0.2% of MZ; E-IV group 0.25 mg/kg OTA + 0.2% of MZ; MZ group supplemented with 0.2% of the adsorbent; and control (K, without feed additive)., Results: Overall, the addition of 0.2% MZ to laying hen feed mitigated the harmful effects of OTA on target organs and reduced the presence of OTA residue in eggs. The groups that received 0.2% of MZ achieved better production results in terms of body weight, number of eggs, and feed consumption, compared to the other treatments., Conclusions: The current findings confirm the efficacy of MZ in preventing performance losses in laying hens exposed to OTA, as well as for improving the welfare and health of food producing animals.

To investigate into the T-2 and HT-2 toxin occurrence, 240 samples of unprocessed cereals (maize, wheat, barley, and oats) were sampled from different fields located in three Croatian regions during 2017-2018. In all samples, sum concentrations of T-2/HT-2 toxin were determined using the ELISA method, while the LC-MS/MS was used as a confirmatory method for both mycotoxins in positive samples (>LOD) and the establishment of T-2 over HT-2 toxin ratios. The results showed oats to be the most contaminated cereal, with T-2/HT-2 toxins detected in 70.0% of samples, followed by barley (40.9%), maize (26.8%) and wheat (19.2%), with the mean T-2/HT-2 ratio ranging from 1:2.7 in maize to 1:4.4 in oats. Sum T-2/HT-2 concentrations in two maize samples were higher than the indicative level recommended by the European Commission, necessitating subsequent investigations into the conditions under which these poorly investigated mycotoxins are produced. Statistically significantly ( p < 0.05) higher concentrations of T-2/HT-2 toxin were determined in oats throughout study regions as compared to those found in wheat, but not maize and barley, while the concentrations of these mycotoxins were related to the regional weather in Croatia.

Characterization of five types of traditional Croatian dry-fermented sausages produced by family farms was performed via identification of superficial mycobiota, physicochemical, sensory, instrumental color, fatty acids & fat quality indices. Detailed characterization of these sausages aimed to achieve standardization of their production and composition and to establish and/or improve their specification protocols. Traditional sausages varied significantly ( p < 0.05) in all analyzed parameters except for the number of mold isolates. Sausages coming from eastern Croatia had a greater mold species diversity, with the highest number of isolated mycotoxigenic species in Slavonian domestic sausage. Sensory evaluation showed good acceptability of all sausages. According to health recommendations, Kulenova Seka showed the most representable values for most of fat quality indices. The results suggest the need for certain modifications in fat & fatty acid composition and, to a lesser extent, in salt content, however not at the expense of product safety, quality and acceptability.

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., Competing Interests: CONFLICT OF INTEREST: None to declare.

In this paper the effect of aflatoxin B 1 , 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.

In this study, the effect of an isolated and well-characterised autochthonous starter culture, glucose and maltodextrin ( w =0.8%) and temperatures of 12 and 20 °C 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. Furthermore, pH value was continuously measured throughout the twenty-day fermentation period. The addition of an autochthonous starter culture and sugars and different fermentation temperatures significantly (p<0.05) affected the instrumental colour and texture parameters of the Slavonian kulen. The fermentation was most intense in the samples with added autochthonous starter culture and 0.8% glucose, and fermented at 20 °C. Microbiological analysis showed that samples with added autochthonous starter culture and fermented at higher temperature contained a higher number of lactic acid bacteria and coagulase-negative staphylococci and were safe. Sensory evaluation confirmed the outcomes of physicochemical and microbiological analyses and showed differences among samples fermented at two different temperatures and with added glucose or maltodextrin and an autochthonous starter culture.

The aim of this study was to evaluate protective effects of vitamin E (50 -150 μM) in ovary cells upon cytotoxic effects induced by two structurally distinct PCB congeners - planar "dioxin-like" PCB 77 and non-planar di-ortho-substituted PCB 153 with an emphasis on identifying differences in the mechanism of vitamin E action depending on the structure of congeners. Application of three bioassays confirmed that PCBs decrease ovarian cell proliferation with slightly profound effects of PCB 77. PCB - induced ROS production and lipid peroxidation were significant for both congeners with also more noticeable effect for PCB 77. Vitamin E pre-incubation has improved viability of cells, reduced ROS formation and lipid peroxidation induced by PCBs' treatment. Preincubation with vitamin E was more effective when cells where treated with non-planar PCB 153. Altogether, vitamin E action was protective, congener specific and more effective when ovary cells were exposed to ortho-substituted PCB congener., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

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.

The aim of this study was to evaluate in vitro toxicity of clenbuterol and its metabolite 4-amino-3,5-dichlorobenzoic acid. Cytotoxicity and pro-oxidative effect of both compounds were studied on human colon adenocarcinoma cell line SW 480. No significant cytotoxic effect of either compound was observed. Results of an Ames test on Salmonella typhimurium did not indicate mutagenic activity of clenbuterol on TA 98 and TA 100 strains, regardless of metabolic activation. Potential mutagenic effects of the highest clenbuterol concentration (2500 ng/ml) were observed on the TA 1535 strain. The obtained results of alkaline comet assay on isolated human lymphocytes suggested that both compounds induced an increase of primary DNA damage in a concentration-dependent manner. 4-ADBA was a slightly more potent inducer of primary DNA damage as compared to clenbuterol. Chromosomal aberration analysis showed that clenbuterol caused a statistically significant increase in the total number of aberrant cells only at the highest concentration tested (3% vs. 0.7% in the negative control). The results of this study might represent a solid frame for designing and planning future studies with both compounds, which should further clarify their mechanisms of action and genotoxic/cytogenetic effects relevant for human risk assessment., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

The aim of the study was to compare the accumulation of β-adrenergic agonist residues clenbuterol (CLB) and salbutamol (SAL) in internal tissues, non-pigmented eyes and hair of laboratory animals repeatedly administered with CLB and SAL during 7 days. Experimental albino guinea pigs (n = 20) were treated with CLB (n = 10) and SAL (n = 10) in anabolic doses of 0.25 and 2.5 mg/kg, whereas the control animal group (n = 10) was left untreated. Methodology validation showed that the ELISA assay to be suitable for β-agonists' semiquantitative determination. The results revealed a significantly higher (P < 0.05) accumulation potential of CLB in comparison with SAL in all investigated tissues. Despite of their lack of pigmentation and the applied dose, the highest residual CLB concentrations were determined in the eyes of the studied animals, followed by their hair, liver, lungs, kidney, heart and adipose and muscle tissue, whereas residual SAL concentrations found in the eyes and hair of the administered animals did not significantly differ (P > 0.05) from those obtained in their internal tissues., (© The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

In the European Union, β(2)-adrenergic agonists like clenbuterol and salbutamol are banned from use as growth promoters. Although clenbuterol and salbutamol both accumulate in the liver, differences in the accumulation rate can be seen among animal species due to different β(2)-adrenoreceptor distributions. The aim of this study was to compare the accumulation of the two in the liver tissue of two different mouse strains. The study included 200 8-week-old BALB/c and C57/BL/6 mice. One group of BALB/c (40) and one group of C57/BL/6 (40) mice were treated with 2.5 mg/kg body mass clenbuterol per os for 28 days. The remaining two animal groups were treated with salbutamol in the same manner. The animals were then randomly sacrificed on day 1, 15 and 30 post treatments. Despite of the same treatment dose, the results revealed clenbuterol to persist in the liver tissue longer than salbutamol. On post treatment day 30, the concentration of clenbuterol residue in C57/BL/6 and BALB/c mice liver tissue were 0.23 ± 0.02 and 0.21 ± 0.03 ng/g, respectively, while residues of salbutamol were not detected. When comparing the accumulation of both compounds between the two mouse strains, it becomes apparent that no significant difference (P > 0.05) in the accumulation rate can be found.

Hair has been shown to be an excellent site for the accumulation of different drugs including β-agonists, and therefore, it would be an appropriate matrix for surveillance for the presence of drug residues. The aim of this study was to determine concentrations and to compare accumulation of two different β-agonists in black and white mice hair by use of ELISA as a screening quantitative method. The study included 200 8-week-old white and black mice. One group of black mice and one group of white mice were treated with clenbuterol in a dose of 2.5 mg/kg body mass per os for 28 days. Other animals were treated in the same way with salbutamol. The highest (±SD) clenbuterol concentration of 631.4 ± 23.5 ng/g in black hair and 228.5 ± 156.2 ng/g in white hair was determined on day 1 of treatment withdrawal. Study results revealed the black-to-white hair ratio of clenbuterol accumulation to be 1:2-1:4 and of salbutamol accumulation 1:1.4. The mean (±SD) salbutamol concentrations determined on day 1 of treatment withdrawal was 23.9 ± 0.9 ng/g and 16.4 ± 1.1 ng/g in black and white hair samples, respectively. The study demonstrated that residues could be determined in hair samples even after a 30-day withdrawal period.

The aim of the present study was to assess the persistence of clenbuterol residues in retinal tissue of pigs after repeated administration in a growth-promoting dose, using enzyme-linked immunosorbent assay (ELISA) as a screening method for quantitative determination. A growth-promoting dose of clenbuterol (20 μg/kg body mass per day) was administered orally to the experimental group (n = 6) for 21 days, whereas control animals (n = 3) were left untreated. Clenbuterol-treated pigs were randomly sacrificed (n = 3) on days 0 and 45 of treatment discontinuation, and clenbuterol residues were determined in retinal tissue dissected from the eye. ELISA was found to be acceptable for quantitative determination of clenbuterol in retinal samples because previous method validation yielded mean recovery values of 84.3-96.5% with variation coefficients < 14%. The mean (± SD) retinal clenbuterol concentration was 1874 ± 114 ng/g immediately upon clenbuterol withdrawal (day 0) and 73 ± 4 ng/g on the last day post-withdrawal (day 45). Study results pointed to a very high potential of clenbuterol accumulation in retinal tissue and marked persistence of clenbuterol residues upon anabolic dose administration, suggesting retinal tissue to be a very useful matrix for effective control of residual clenbuterol in food-producing pigs.