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Visual examination of visually recognisable substances, including microscopy, focus on targets or contaminants such as particles of animal origin, plant seeds, spore bodies of moulds, sclerotia, packaging material, microplastic and 'Besatz' (everything that differs from the norm). The two principal results are counts (numbers) and weights for macroscopic methods, or presence/absence for microscopic methods. The level of detection equals at least the size of one unit, usually with a weight exceeding 1 mg, which is in the range of parts per million (ppm). These parameters do not follow a normal distribution but Poisson (counts), lognormal (weights) or binomial (Booleans) distributions, with effect on the interpretation of validation parameters. As for other domains, examination methods for visual monitoring need to be properly validated and quality control during actual application is needed. In most cases procedures for validation of visual methods are based on principles adopted from other domains, such as chemical analysis. A series of examples from publications show inconsistent or not correct implementations of these validation procedures, which stress the need for dedicated validation procedures. Identification of legal ingredients and composition analysis in the domain of visual examination relies on the expertise of the laboratory staff, therefore validation of a method usually includes the validation of the expert. In the view of these specific circumstances, a Guidance for quality assurance and control of visual methods has been developed, which are being presented and discussed in this paper. The general framework of the Guidance is adopted from ISO standards (17023, 17043, 13528). Part 1 of the Guidance includes the general background, theory and principles. Part 2 presents the actual validation procedures with experimental designs and equations for calculating the relevant parameters, and can be used as blueprint for a SOP in a quality management system. An EURL and NRL network for physical hazards is strongly recommended.

Slavonska kobasica je trajna kobasica koje se u seoskim domaćinstvima u Slavoniji proizvodi tradicionalnom tehnologijom, pri kojoj se tijekom zrenja na ovitku proizvoda mogu razviti plijesni. Cilj je ovog rada bio identificirati površinske plijesni i utvrditi njihov utjecaj na kvalitetu i zdravstvenu sigurnost proizvoda, uključujući pojavnost mikotoksina okratoksina A (OTA) i aflatoksina B1 (AFB1). Tijekom rada komunicirano je sa šest proizvodnih lokaliteta na području Slavonije. Pripremljeno je 18 uzoraka slavonske kobasice koje su analizirane na kraju proizvodnog procesa u trajanju od tri mjeseca. Ukupno je identificirano 8 različitih vrsta plijesni, među kojima je dominirao rod Penicillium (88,89 %). Osim šest Penicillium vrsta, izolirana je po jedna vrste iz rodova Aspergillus (8,33 %) i Mucor (2,78 %). Identificirani su i mogući izvori OTA i AFB1, Penicillium verrucosum i Aspergillus flavus, a određena je samo prisutnost OTA i to u koncentraciji od 5,10 μg/kg., Slavonian sausage is a cured sausage produced on rural holdings using traditional technology, in which moulds are permitted to develop on the product surface during the maturation period. The purpose of this study was to identify the surface moulds and determine their influence on product quality and safety, including the occurrence of the mycotoxins ochratoxin A (OTA) and aflatoxin B1 (AFB1), from six production facilities in the Slavonia region. A total of 18 samples of Slavonian sausage were prepared and analysed at the end of the production process, i.e., after three months. A total of eight species of mould were identified, and the dominant genus was Penicillium (88.89%) with six species, one species from the genus Aspergillus (8.33%) and one species from the genus Mucor (2.78%). Possible causative agents of OTA and AFB1 were identified, Penicillium verrucosum and Aspergillus flavus, though only the presence of OTA was confirmed in a concentration of 5.10 μg/kg.

Mikotoksini predstavljaju prirodne toksine koji učestalo kontaminiraju hranu i hranu za životinje, a primarno ih sintetiziraju plijesni iz rodova Aspergillus, Penicillium, Fusarium, Alternaria i Claviceps. Ove tvari predstavljaju vrlo stabilne spojeve, otporne na različite uvjete proizvodnje, prerade i skladištenja hrane. Mikotoksini mogu prouzročiti niz štetnih učinaka po zdravlje ljudi i životinja, tzv. mikotoksikoze, uključujući karcinogenost, mutagenost, genotoksičnost, imunotoksičnost, neurotoksičnost, hepatotoksičnost, nefrotoksičnost, te druge, pri čemu često istovremeno djeluju na više ciljnih mjesta u organizmu, u ovisnosti o toksičnosti samog mikotoksina te dozi i vremenu izloženosti. Za veliki broj mikotoksina još uvijek nema dovoljno dokaza o njihovim štetnim učincima na ljudsko zdravlje, ali se temeljem utvrđene osjetljivosti različitih životinjskih vrsta pretpostavlja da i u ljudi mogu izazvati slična djelovanja. Ujedno, za većinu mikotoksina koji se učestalo zajednički pojavljuju u hrani i hrani za životinje nisu razjašnjeni njihovi učinci u organizmu koji mogu varirati od antagonističkih do sinergističkih. Ovaj rad daje pregled mehanizama djelovanja mikotoksina od najvećeg javnozdravstvenog značaja i njihovih toksičnih učinaka u organizmu.

Kvasci su česti zagađivači mliječnih proizvoda, međutim, koriste se i kao starter kulture za poboljšavanje svojstava finalnog proizvoda. Njihova prisutnost u povećanom broju može prouzročiti nepoželjne promjene mliječnih proizvoda, kao što su neugodan miris, okus i izgled. Klasična identifikacija kvasaca na temelju morfoloških i biokemijskih svojstava spora je i nepouzdana. Cilj rada bio je identificirati kvasce iz 30 uzoraka mlijeka i svježeg sira te ustvrditi pripadaju li izolirani kvasci u poželjne ili nepoželjne vrste, odnosno predstavljaju li potencijalni rizik po zdravlje ljudi. Izolirani kvasci identificirani su sekvenciranjem D1/D2 regije gena 28 S rRNK. Dobivenim sekvencama ustvrđeno je da izolirani kvasci pripadaju vrstama: Kluyveromyces marxianus, Candida tropicalis, Trichosporon coremiiforme, Rhodotorula mucilaginosa, Trichosporon ovoides, Pichia kudriavzevii, Issatchenkia orientalis, koje se smatraju kvascima zagađenja te Debaryomyces hansenii vrsti koja se može koristiti i kao starter kultura. S obzirom da kvasci nisu termorezistentni mikroorganizmi, trebali bi biti uklonjeni tijekom pasterizacije, no u tradicionalnom načinu proizvodnje sira mlijeko se termički ne obrađuje, stoga je povećan rizik od pojavnosti kvasaca u finalnom proizvodu. Izolirane se vrste vrlo često nalaze i u okolišu i/ili na ljudima, odnosno životinjama. Stoga se nameće zaključak kako se glavnina kontaminacije kvascima događa kao posljedica neadekvatne higijenske prakse.

Sterigmatocistin je mikotoksin kojeg sintetiziraju uglavnom plijesni iz roda Aspergillus te u manjoj mjeri iz rodova Emericella, Chaetomium, Humicola i Botryotrichum. Ovaj mikotoksin je prekursor u biosintezi aflatoksina, njemu slične kemijske strukture, pa su tako i toksični učinci sterigmatocistina slični onima aflatoksina B1. Sterigmatocistin pokazuje brojne toksične učinke u organizmu, uključujući karcinogeno i mutageno djelovanje, te je i Međunarodna agencija za istraživanje raka (engl. International Agency for Research on Cancer, IARC) sterigmatocistin uključila u skupinu 2B potencijalnih karcinogena za ljude. Podaci o prisutnosti sterigmatocistina u hrani i hrani za životinje su ograničeni, te je od strane Europske agencije za sigurnost hrane (eng. European Food Safety Authority, EFSA), s ciljem procjene rizika izloženosti ljudi ovom mikotoksinu, dana preporuka za prikupljanje podataka o njegovoj pojavnosti uz primjenu visokoosjetljivih analitičkih metoda. Sterigmatocistin je do sada pronađen u hrani za životinje, žitaricama, kruhu, orašastim plodovima, zrnima kave, začinima, pivu te siru. Kako bi se spriječila kontaminacija namirnica te negativni učinci sterigmatocistina na zdravlje ljudi i životinja, jedan od ključnih čimbenika predstavlja prevencija rasta toksikotvornih plijesni.

Ergot alkaloids (EAs) are mycotoxins produced by several species of fungi of the genus Claviceps, among which Claviceps purpurea is the most widespread in Europe. This species has been found in many economically important cereal grains, such as rye, wheat, triticale, barley, millet and oats. The distribution of EA contamination has a sporadic incidence, with many factors involved in its occurrence, greatly varying between fungal strains, geographic regions, host plants and regional/local weather conditions. Cool, damp weather favours ergot by enhancing the germination of sclerotia. The aim of this study was to investigate the occurrence of ergot sclerotia and EAs in wheat and rye grain samples (n = 64) collected during 2021 from Croatian cereal producers in central and eastern Croatia. In two rye samples, the presence of ergot sclerotia was detected in the amount of 259 mg/kg and 536 mg/kg, whereas no wheat samples tested positive for ergot sclerotia. A higher contamination with EAs was determined in the rye samples (18% contaminated; max 167.4 μg/kg), while a lower frequency of contamination was determined in wheat, with only one positive sample (1.9%; 68.5 μg/kg). The results indicate low-level EA contamination of wheat and rye cultivated by Croatian producers during the study period. However, despite the low incidence of positive rye samples with EAs, the contents of ergot sclerotia in two samples were higher than permitted by the legislation for foodstuffs. Since the levels of these mycotoxins and ergot sclerotia content can vary depending on a number of factors, further research is required over a longer period of time and under different cereal cultivation and processing conditions., Ergot-alkaloidi (EA) su mikotoksini koje proizvodi nekoliko vrsta gljivica iz roda Claviceps, među kojima je i Claviceps purpurea najraširenija gljivica u Europi. Ova vrsta je pronađena u ekonomski važnim žitaricama, kao što su: raž, pšenica, tritikal, ječam, proso i zob. Distribucija kontaminacije EA ima sporadičnu pojavu s mnogo čimbenika koji su uključeni u njihovu pojavu, uvelike varirajući između sojeva gljivica, geografskih regija, biljaka domaćina i regionalnog vremena, što ukazuje da hladno i vlažno vrijeme pogoduje ergotu tako što potiče klijanje sklerocija. Cilj je ovog istraživanja bio istražiti pojavu ergot- sklerocija i EA u uzorcima pšenice i raži (n=64) uzorkovanih tijekom 2021. godine od hrvatskih proizvođača žitarica u središnjoj i istočnoj Hrvatskoj. U dva uzorka raži utvrđena je prisutnost ergot-sklerocija u količini od 259 mg/kg i 536 mg/kg. Niti u jednom uzorku pšenice nije pronađen ergot-sklerocij. Veća kontaminacija s EA utvrđena je u uzorcima raži (18 % kontaminirano, max 167,4 μg/kg), dok je manja učestalost kontaminacije, sa samo jednim pozitivnim uzorkom (1,9 %), utvrđena u pšenici (68,5 μg/kg). Rezultati su ukazali na nisku razinu kontaminacije s EA pšenice i raži koje su uzgojili hrvatski proizvođači tijekom cijelog istraživanog razdoblja. Međutim, iako je utvrđen nizak postotak pozitivno testiranih uzoraka raži na EA, sadržaj ergot-sklerocija u dva uzorka bio je veći nego što je to za hranu zakonodavstvom dopušteno. Budući da razine tih mikotoksina i ergot-sklerocija mogu varirati ovisno o brojnim čimbenicima, potrebno je njihovo daljnje istraživanje tijekom dužeg vremenskog razdoblja i pod različitim uvjetima uzgoja i obrade žitarica.

By increasing the resistance of seeds against abiotic and biotic stress, the possibility of cereal mold contamination and hence the occurrence of secondary mold metabolites mycotoxins decreases. The use of biological methods of seed treatment represents a complementary strategy, which can be implemented as an environmental-friendlier approach to increase the agricultural sustainability. Whereas the use of resistant cultivars helps to reduce mold growth and mycotoxin contamination at the very beginning of the production chain, biological detoxification of cereals provides additional weapons against fungal pathogens in the later stage. Most efficient techniques can be selected and combined on an industrial scale to reduce losses and boost crop yields and agriculture sustainability, increasing at the same time food and feed safety. This paper strives to emphasize the possibility of implementation of biocontrol methods in the production of resistant seeds and the prevention and reduction in cereal mycotoxin contamination.

Mycobiota grow on the surface of traditional meat products (TMP), which gives them a specific texture, flavour, and colour. However, in unfavourable conditions moulds produce mycotoxins if they possess the genes involved in their biosynthesis. The most toxic mycotoxins in TMP products are aflatoxin B1 (AFB) and ochratoxin A (OTA), while the incidence of cyclopiazonic (CPA) acid and citrinin (CIT) on meat products has until now not been researched. The aim of this study was to identify moulds species capable of producing the studied mycotoxins by proving the existence of the genes involved in their biosynthesis. This study analysed 250 TMPs sampled during 2020 and 2021 from five different climate regions of the Republic of Croatia (eastern, northern, central, western, and southern). The tested genes involved in the AFB1 biosynthesis are: aflD encoding norsolorinic acid reductase, aflM encoding versicolorin A dehydrogenase and aflP encoding sterigmatocystin-Omethyltransferase, OTA otanpsPN encoding OTA nonribosomal peptide synthetase and otapks encoding OTA polyketide synthase ; CIT pksCT gene encoding CIT polyketide synthase, and CPA dmaT encoding dimethylalyl tryptophan synthase. A total of 181 isolates of 9 different potentially toxic mould species were identified, out of which 50 % of the identified isolates of potential AFB1 producers (A. flavus), 58 % isolates of potential OTA producers (P. nordicum, A. niger, A. westerdijkiae, A. welwitschie, A. tubingensis), 71 % of isolates of potential CIT producers (P. citrinum) and 48 % of isolates of potential CPA producers (P. commune, A. flavus, P. polonicum) all had the investigative genes involved in mycotoxin biosynthesis. In 21 % of the samples contaminated with some of the detected mycotoxins (OTA or CPA), their producers (P. nordicum, P.commune and A. flavus) were determined, while contamination of the samples in which the producers were not identified can be explained either by some previous contamination of TMP ingredients or by the possibility that the producers had been covered and became overgrown by other mould species at the end of the ripening stage and therefore avoided detection. The incidence of the number and identified mould species has been linked to data on the weather conditions relating to the various Croatian regions. Therefore, the western region had the highest number of isolated toxic mould species, followed by the northern region, while the greatest diversity of toxic mould species was in the western and eastern regions.

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.

This study aimed to identify surface mould species overgrowing the Croatian protected meat products “Istarski pršut” and “Dalmatinski pršut” and their effect on sensory properties. Dry-cured hams were produced in 2018/2019 and obtained from annual fairs. The predominant surface species found on “Dalmatinski pršut” were Aspergillus chevalieri, Penicillium citrinum and Aspergillus cibarius, whereas those overgrowing “Istarski pršut” were Aspergillus proliferans, P. citrinum and Penicillium salamii. The results show species diversity, higher presence, and greater variety of Aspergillus species in “Dalmatinski pršut” in comparison to “Istarski pršut”, and significant variations in 9 of 20 sensory attributes. Principal component analysis revealed a clear distinction between the two, and a large contribution of P. salamii and Penicillium bialowienzense to one principal component. The texture traits, smoky odour, muscle and subcutaneous fatty tissue colour, and mould species found are valuable for product characterisation. The results also indicate that mould species may be responsible for some sensory traits, such as tenderness, juiciness, and lesser freshness.

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 B1 (AFB1), 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 AFB1 and its producers were not detected in any of the analysed samples (&lt, LOD).

Documented cases of mycotoxin occurrence in meat products call for further research into potential contamination sources, especially given an ever more increasing consumption of these nutritionally rich products. These foodstuffs can be contaminated with mycotoxins through three pathways: contaminated spices and other raw materials, mycotoxin-producing moulds present on the surface of dry-cured meat products, and carry-over effect from farm animals exposed to contaminated feed. In order to establish meat products’ mycotoxin contamination more precisely, the concentrations of all mycotoxins of relevance for these products should be determined. This manuscript reviews data on major mycotoxins present in different types of meat products, and discusses the contamination pathways, contamination levels and control & preventative measures.

Fermented sausages have been manufactured traditionally in the Mediterranean basin. Istrian sausages are one of the many regionally specific products distinguished by the specific traditional recipes and consequentially exceptional sensory characteristics. The aim of this study was to evaluate sensory, physico- chemical and microbiological changes occurring during production and ripening of Istrian sausages made at traditional households, and to compare the results obtained with the identical product produced in industrial conditions. Additional goal was to characterise the autochthonous lactic acid bacteria populations and to obtain the candidate starter strains. Production stages and product ripening parameters could be strictly defined in controlled industrial conditions of the production process, and the results of this study can serve to standardise the production of traditional Istrian sausages. The results from sensory and physico- chemical examination do not show differences in relation to sausages of a similar type investigated in other papers. Lactic acid bacteria were favoured during fermentation at rising salt concentrations, consisting mostly of Lactobacillus strains followed with the abundance of coagulase-negative cocci. Further, we targeted the autochthonous lactic LAB population to select the candidate starter strains for controlled sausage fermentation. Fifty Gram-positive strains with catalase-negative phenotype, rod-shape or cocci morphology, effective acidification kinetics and production of lactic acid identified by sequence analysis of 16S rRNA were considered LAB. The selected LAB strains were able to grow in a range of pH values and high salt concentration important for their metabolic activity during sausage fermentation. According to our results, Lactobacillus sakei, which is a prevalent strain during fermentation of various types of fermented sausages, was frequently detected. Lactobacillus plantarum and Lactobacillus curvatus were also detected in abundance, implying their possible competitiveness among the complex microbial population. Isolated autochthonous strains L. curvatus S30, L. sakei S32 and L. plantarum S50 produced high lactic acid concentrations, tolerated NaCl concentrations present during sausage fermentation, survived at high concentrations of hydrogen ions and inhibited six pathogens associated with contamination of meat products in a strain-specific manner.

During the curing period of dry-fermented sausages, the surface mycoflora changes according to the environmental conditions and intrinsic factors, such as nutrient content, water activity, pH value and the presence of antimicrobial substances (starter cultures). In this study, 25 samples of dry-fermented sausages from Croatian family small holdings were examined, the casings peeled off and examined for mycobiota before and after six months of refrigerated storage in plastic bags, at +4 °C in darkness, at a low humidity and evaporation rate. For mycobiota identification, traditional (macroscopic and microscopic) and molecular (Polymerase Chain Reaction, PCR) methods were combined. At the beginning of the study (after sausage curing), the prevalent isolated species were from the genera Penicillium (74%), Aspergillus 14% and Mucor (12%). After the six month period, a different mould structure was observed, with the dominant isolation of Aspergillus genus (64%), followed by the genera Penicillium (34%) and Mucor (2%). In comparison with the literature data, it can be concluded that the casing mycoflora on dry-fermented sausages stored in the refrigerator changed in the same way as if they not been refrigerated, though the process of mycoflora change was slower with refrigeration., Tijekom razdoblja zrenja trajnih fermentiranih kobasica mikoflora koja obrasta i prekriva njihovu površinu mijenja se u ovisnosti o uvjetima okoliša i “unutarnjim čimbenicima”, koji uključuju sadržaj hranjivih sastojaka, aktivitet vode, pH vrijednost i prisutnost antimikrobnih tvari (starter kulture). U ovom je istraživanju uzorkovano dvadeset i pet parova trajnih fermentiranih kobasica s malih hrvatskih obiteljskih poljoprivrednih gospodarstava, odljušten je njihov ovitak te je ispitivana mikoflora prije i nakon skladištenja u hladnjaku tijekom šest mjeseci, u plastičnim vrećicama, na +4 °C, u tami, pri maloj vlažnosti i brzini isparavanja. Za identifikaciju plijesni kombinirane su tradicionalne (makroskopske i mikroskopske) i molekularne (lančana reakcija polimeraze, PCR) metode. Na početku istraživanja (nakon sazrijevanja i uzimanja uzoraka kobasica) prevladavale su izolirane vrste iz roda Penicillium (74 %), Aspergillus 14 % i Mucor (12 %). Nakon razdoblja od šest mjeseci uočena je drugačija struktura plijesni, pri čemu je pretežno izoliran rod Aspergillus (64 %), a zatim Penicillium (34 %) i Mucor (2 %). Sagledavajući literaturne podatke može se zaključiti da se mikoflora na ovitcima trajnih fermentiranih kobasica pohranjenih u hladnjaku mijenjala na isti način kao što bi se i mijenjala bez pohranjene u hladnjaku, jedino što bi proces promjena mikoflore vremenski trajao kraće.

Cyclopiazonic acid (CPA) is a mycotoxin produced by several moulds of the Aspergillus and the Penicillium genus. Insofar, CPA has been detected in various foods of plant origin, such as peanut, corn, figs, rice, wheat and tomato, but also in food of animal origin, such as cheese, milk and ham. CPA levels in different types of dried meat products are generally unexplored, analytical methods suitable for CPA detection thereby being limited. In view of the above, the objectives of this study were to develop a sensitive analytical method of CPA detection in dry-fermented meat products, and to employ this method in the analysis of traditional dry-fermented sausages. The developed analytical method had the LOD (Limit of Detection) of 2.17 μg/kg and the LOQ (Limit of Quantification) of 7.15 μg/kg. The recoveries ranged from 95.49% to 102.05% with the linearity of R2 = 0.996, while the matrix effect in terms of ion enhancement was almost negligible (1.01%). The analyses of 47 traditional dry-fermented sausages revealed CPA presence in 7 samples, in the concentration ranging from 2.55 μg/kg to 59.80 μg/kg. The presented method can be further used for the analysis of other types of dry-fermented meat products, especially those longer ripen, on which a mould growth can be substantial.

This study aimed to identify surface mould species overgrowing the Croatian protected meat products “Istarski pršut” and “Dalmatinski pršut” and their effect on sensory properties. Dry-cured hams were produced in 2018/2019 and obtained from annual fairs. The predominant surface species found on “Dalmatinski pršut” were Aspergillus chevalieri, Penicillium citrinum and Aspergillus cibarius, whereas those overgrowing “Istarski pršut” were Aspergillus proliferans, P. citrinum and Penicillium salamii. The results show species diversity, higher presence, and greater variety of Aspergillus species in “Dalmatinski pršut” in comparison to “Istarski pršut”, and significant variations in 9 of 20 sensory attributes. Principal component analysis revealed a clear distinction between the two, and a large contribution of P. salamii and Penicillium bialowienzense to one principal component. The texture traits, smoky odour, muscle and subcutaneous fatty tissue colour, and mould species found are valuable for product characterisation. The results also indicate that mould species may be responsible for some sensory traits, such as tenderness, juiciness, and lesser freshness.

Morphological characteristics of whale bones (Order: Cetacea) have shapedas a result of adaptation to life in the aquatic environment and whale diving habits. This study deals with the postnatal bone development and microscopic characteristics of bones of bottlenose dolphins (Tursiops truncatus). Bone sections of the right humerus and the fifth right rib of 106 dolphins aged less than 1 to 21 years were used to determine the density of the primary, secondary and remodelling osteons, the magnitude of the area occupied by the osteons, and the number of osteon lamellae. Bone sections were analysed using a compound microscope with standard and polarized light. Our study showed the primary osteons of the humerus and the rib of the bottlenose dolphin to occupy a smaller area than the secondary ones. The secondary osteon area in the humerus is significantly larger than that in the rib. All rib osteons are significantly denser than those of the humerus. In both bones, the number of osteon lamellae is higher in secondary than in primary osteons. The number of secondary and remodelling osteons and the areas occupied by them increase with age, as oppose to primary osteons in which a decrease was noted. The number of lamellae in primary osteons decreases with age, while in secondary osteons their number increases. Within the study frame, morphometric osteon values of the bottlenose dolphin were determined, to the best of our knowledge, for the first time everas concerns the aquatic mammals.

Kvasci su česti zagađivači mliječnih proizvoda, međutim, koriste se i kao starter kulture za poboljšavanje svojstava finalnog proizvoda. Njihova prisutnost u povećanom broju može prouzročiti nepoželjne promjene mliječnih proizvoda, kao što su neugodan miris, okus i izgled. Klasična identifikacija kvasaca na temelju morfoloških i biokemijskih svojstava spora je i nepouzdana. Cilj rada bio je identificirati kvasce iz 30 uzoraka mlijeka i svježeg sira te ustvrditi pripadaju li izolirani kvasci u poželjne ili nepoželjne vrste, odnosno predstavljaju li potencijalni rizik po zdravlje ljudi. Izolirani kvasci identificirani su sekvenciranjem D1/D2 regije gena 28 S rRNK. Dobivenim sekvencama ustvrđeno je da izolirani kvasci pripadaju vrstama: Kluyveromyces marxianus, Candida tropicalis, Trichosporon coremiiforme, Rhodotorula mucilaginosa, Trichosporon ovoides, Pichia kudriavzevii, Issatchenkia orientalis, koje se smatraju kvascima zagađenja te Debaryomyces hansenii vrsti koja se može koristiti i kao starter kultura. S obzirom da kvasci nisu termorezistentni mikroorganizmi, trebali bi biti uklonjeni tijekom pasterizacije, no u tradicionalnom načinu proizvodnje sira mlijeko se termički ne obrađuje, stoga je povećan rizik od pojavnosti kvasaca u finalnom proizvodu. Izolirane se vrste vrlo često nalaze i u okolišu i/ili na ljudima, odnosno životinjama. Stoga se nameće zaključak kako se glavnina kontaminacije kvascima događa kao posljedica neadekvatne higijenske prakse., Yeasts are a common contaminant of dairy products, but they can also be used as a starter culture to improve the characteristics of the final product. Their presence in an increased number causes undesirable changes in dairy products, such as an unpleasant appearance, acrid smell and foul taste. The classical identification of yeasts based on morphological and biochemical properties is slow and unreliable. The aim of this study was to identify isolated yeasts from 30 dairy products, and to determine if they belong to desired or undesirable species. Isolated yeasts were identified by sequencing the D1/D2 domain of the gene 28S rRNA. The obtained sequences determined that the isolated yeasts belong to the species: Kluyveromyces marxianus, Candida tropicalis, Trichosporon coremiiforme, Rhodotorula mucilaginosa, Trichosporon ovoides, Pichia kudriavzevii, Issatchenkia orientalis, which are considered contaminants, and Debaryomyces hansenii, a species that can be used as a starter culture. Since yeasts are not thermoresistant microorganisms, they should be removed during pasteurization. However, milk is not heat-treated in traditional cheese-making production, thus increasing the risk of yeast in the final product. These isolated species are very often found in the environment and/or on humans and animals therefore, it can be concluded that the majority of yeast contamination occurs as a result of inadequate hygiene.

Tijekom posljednjeg desetljeća znanstvenici sve više ukazuju na globalno zagrijavanje, dovodeći klimatske promjene u poveznicu s novom strukturom vrsta plijesni producenata mikotoksina po geografskim područjima svijeta. Izraženije klimatske promjene mogu u budućnosti izmijeniti otpornost domaćina i interakciju domaćin-patogen te imati znatan utjecaj na razvoj toksikogenih plijesni i produkciju mikotoksina kao njihovih sekundarnih metabolita. Novonastala klimatološka situacija izaziva sve veću pozornost i zahtijeva nove dijagnostičke alate, kao i nove spoznaje o biološkim karakteristikama poljoprivrednih kultura koje se uzgajaju i genetici toksikogenih plijesni. Zbog sve povoljnijih klimatskih uvjeta za rast Aspergillus flavus na području Europe, u narednom razdoblju postoji povećani rizik od onečišćenja žitarica izrazito toksičnim aflatoksinima. Ujedno, profil mikotoksigenih vrsta Fusarium počinje se sve znatnije mijenjati, a posebno je zabrinjavajući porast onečišćenja s F. graminearum u sjevernom i središnjem dijelu Europe. S obzirom na očigledne klimatske promjene, potrebna su daljnja istraživanja koja uključuju „prediktivne modele“ s više varijabli, a koji bi u bliskoj budućnosti omogućili kvalitetni pristup strategiji prevencije kontaminacije toksikotvornim plijesnima i mikotoksinima u hrani i hrani za životinje., Over the last decade, scientists have increasingly pointed to global warming, linking climate change to a new structure of mycotoxin-producing mould species across the world. In the future, more pronounced climate changes may alter host resistance and host-pathogen interaction, and have a significant impact on the development of toxicogenic moulds and the production of mycotoxins as their secondary metabolites. The newly emerging climatological situation is attracting increasing attention, demanding new diagnostic tools and new insights into the biological characteristics of cultivated crops and the genetics of toxicogenic moulds. In Europe, the climate conditions are highly favourable for the growth of Aspergillus flavus, bringing increased future risk of contamination of cereals with highly toxic aflatoxins. Meanwhile, the profile of mycotoxigenic species of Fusarium is beginning to change significantly, and increased reports of contamination with F. graminearum in northern and central Europe are particularly worrisome. Given the obvious climate change, further research is needed that will include predictive models with several variables, to provide an appropriate future approach to the strategy of preventing contamination by toxic moulds and mycotoxins in food and feed.

Since adulteration can have serious consequences on human health, it affects market growth by destroying consumer confidence. Therefore, authentication of food is important for food processors, retailers and consumers, but also for regulatory authorities. However, a complex nature of food and an increase in types of adulterants make their detection difficult, so that food authentication often poses a challenge. This review focuses on analytical approaches to authentication of food of animal origin, with an emphasis put on determination of specific ingredients, geographical origin and adulteration by virtue of substitution. This review highlights a current overview of the application of target approaches in cases when the compound of interest is known and non-target approaches for screening issues. Papers cited herein mainly concern milk, cheese, meat and honey. Moreover, advantages, disadvantages as well as challenges regarding the use of both approaches in official food control but also in food industry are investigated.

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

Mycotoxins may contaminate food of animal origin due to the carry-over effect and represent a potential risk to human health. The problem of Fusarium mycotoxin contamination becomes an issue especially during rainy years characterised by substantial temperature changes. The aim of this study was to investigate into the level of Fusarium mycotoxins zearalenone (ZEN), deoxynivalenol (DON) and fumonisins (FUM) in maize silage (n=21), concentrated dairy cattle feeds (n=56) and cow milk samples (n=105), taken during 2015 from households located in four Croatian regions. The presence of mycotoxins was determined using validated ELISA methods. A high level of feedstuffs’ contamination was evidenced, especially with ZEN, with values higher than recommended observed in 9.5 % of maize silage samples. Fourteen point three percent (14.3 %) of milk samples were DON positive, with the toxin concentrations ranging from 5.4 to 67.3 μg/L. ZEN was determined in 94.3 % of milk samples, ranging from 0.3 to 88.6 μg/L. FUM were not detected in any of the analysed milk samples. Given the tolerable daily intakes (TDIs) defined for these mycotoxins, human health risks arising from the consumption of cow milk can generally be considered low, even in times characterised by weather conditions that facilitate the production of Fusarium mycotoxins in cereals subsequently used as dairy cattle feed. The exception represents particular milk samples in which high ZEN concentrations were found.

This study investigated the surface-to-interior mycotoxin contamination of the regional speciality meat product ‘slavonski kulen’ that occurred during a 12-month period and identified moulds present on its surface during the final production stage. In total, 15 pieces of slavonski kulen were produced and sampled at production months 0, 3, 6, 9 and 12 (three samples per piece). Concentrations of aflatoxin B1 (AFB1) and ochratoxin A (OTA) were determined using the ELISA method, while the identification of moulds made use of both traditional and molecular methods. A 12-month- long production of dry-fermented sausages under controlled conditions that did not include product cleaning at any point caused significant mycotoxin contamination, with determined mean AFB1 and OTA concentrations of 11.79 ± 2.34 and 16.13 ± 3.32 μg kg–1, respectively. Significantly higher mycotoxin levels were observed in the external layers of the products in comparison with their interior and, even more so, central parts. In total, five Penicillium species (P. polonicum, P. terrigenum, P. solitum, P. jugoslavicum and P. corylophilum) and two Aspergillus species (A. versicolor and A. sydowii) were isolated. Moulds responsible for the production of AFB1 and OTA in the later production stages had probably been overgrown by non- toxicogenic moulds, so that the former did not last until the end of the sausages’ production process and were therefore not identified.

Cilj rada je istražiti mikrobiološku kvalitetu uzoraka meda, uzorkovanih na području sjeverozapadne Hrvatske. Tijekom 2018. godine uzorkovano je ukupno 40 uzoraka različitih vrsta meda te je provedena analiza aerobnih mezofilnih bakterija, bakterija iz porodice Enterobacteriaceae, sulfitreducirajućih klostridija te kvasaca i plijesni prema kriterijima Vodiča za mikrobiološke kriterije za hranu. Od ukupnog broja pretraženih uzoraka, prisutnost kvasaca utvrđena je u 25,0% uzoraka meda, dok je u 27,5% uzoraka utvrđena prisutnost plijesni u koncentraciji od 10 do 100 cfu/g. Na izdvojenim izolatima plijesni koji su bili nesukladni u odnosu na preporučeni kvantitativni kriterij (10 cfu/g), provedena je daljnja identifikacija, čime je u uzorcima meda utvrđena prisutnost sedam rodova plijesni: Cladosporium, Penicillium, Alternaria, Mucor, Acremonium, Paecylomyces i Pestalotiopsis. Iako se rodovi plijesni izdvojeni u ovom istraživanju smatraju čestim onečišćivačima pčelinjih proizvoda, njihova prisutnost upućuje na nužnost proizvodnje sukladno pravilima dobre pčelarske prakse uz sustavno provođenje mikrobiološke kontrole meda i drugih pčelinjih proizvoda., The aim of this study was to examine the microbiological quality of honey sampled in the northwest region of Croatia. During 2018, a total number of 40 honey samples were taken and analysed for the number and/or presence of aerobic mesophilic bacteria, bacteria from the Enterobacteriaceae family, sulphite- reducing clostridia, yeasts and moulds, according to Croatian National Guidelines on microbiological criteria for foodstuffs. Out of the total number of analysed honey samples, the presence of yeasts was noted in 25% of the samples, whereas moulds were determined in 27.5% of the samples, ranging from 10 to 100 cfu/g. Isolated moulds that were unsuitable according to the recommended quantitative criterion (10 cfu/g) were singled out and subjected to further identification, whereby seven genera of moulds were recovered: Cladosporium, Penicillium, Alternaria, Mucor, Acremonium, Paecylomyces and Pestalotiopsis. Although the majority of moulds isolated in this study are common contaminants of bee products, their presence indicates the need for production according to the guidelines for good beekeeping practices with systematic microbiological controls of honey and other bee products.

Mikotoksini predstavljaju toksične tvari koje učestalo kontaminiraju hranu i hranu za životinje, uzrokujući štetne učinke na zdravlje ljudi i životinja te značajne ekonomske gubitke diljem svijeta. Ovaj rad daje pregled dosadašnjih istraživanja i spoznaja o metodama biološke detoksifikacije mikotoksina. Biološka detoksifikacija podrazumijeva primjenu mikroorganizama i njihovih enzima s ciljem razgradnje ili modifikacije strukture mikotoksina, a rezultira transformacijom izvornih spojeva i nastankom manje toksičnih produkata, odnosno dekontaminacijom mikotoksina. Međutim, dosadašnja istraživanja i dobivene spoznaje još uvijek nisu rezultirale primjenom u prehrambenoj i stočarskoj industriji odnosno odobravanjem njihove uporabe od strane zakonodavstva. Potrebna su daljnja istraživanja o mehanizmima biorazgradnje, sigurnosti mikroorganizama, odnosno utjecaju primjene bioloških metoda na nutritivna svojstva i nastale produkte degradacije, kako bi metode biološke detoksifikacije mikotoksina mogle biti učinkovito primjenjene u industriji.

Tijekom dugotrajnog zrenja površinu trajnih tradicionalnih mesnih proizvoda (TMP) uobičajeno obrastaju plijesni. Među njima, toksikotvorne vrste produciraju sekundarne metabolite mikotoksine, koji imaju izrazite štetne posljedice po zdravlje ljudi i životinja. Aflatoksini i okratoksini imaju dokazane karcinogene i mutagene učinke u organizmu, a njihova pojavnost u TMP najčešće je posljedica prekomjernog rasta površinske plijesni. Osim ovim izravnim putem, mikotoksini u TMP mogu biti prisutni i neizravno, putem kontaminirane stočne hrane, čija uporaba na farmskim životinjama rezultira prijenosom (carry-over efektom) u finalne mesne proizvode. Zbog učestalosti kontaminacije važan je razvoj potvrdnih analitičkih metoda za detekciju mikotoksina te primjena metoda za identifikaciju površinskih plijesni. Kako bi se spriječila kontaminacija TMP i mogući štetni učinci po zdravlje potrošača, potrebno je poduzeti sve preventivne radnje i provoditi sustavnu kontrolu ovih kontaminanata. Ovaj rad daje pregled svojstava aflatoksina i okratoksina kao mikotoksina od najvećeg značaja za pojavnost u mesnim proizvodima, plijesni koje ih produciraju te metoda detekcije, prevencije i redukcije kontaminacije TMP., During long-lasting ripening, the surface of dry traditional meat products (TMP) is usually owergrown by moulds. Among them, toxicogenic species produce mycotoxins as secondary metabolites, which have significant adverse effects on human and animal health. Aflatoxins and ochratoxins have proven carcinogenic and mutagenic effects in the body, and their occurrence in TMP is the most common consequence of overexpression of surface moulds. In addition to this direct route, mycotoxins in TMP may also be present indirectly through contaminated animal feed, whose use in farm animals results in carry-over effects in final meat products. Due to the frequency of contamination, development of confirmatory analytical methods for mycotoxin detection and the application of surface moulds identification methods is important. To prevent TMP contamination and possible adverse effects on consumer health, all preventive actions should be taken and systematic control of these contaminants should be undertaken. This paper gives an overview of the properties of aflatoxins and ochratoxins as mycotoxins of the greatest importance for the occurrence of meat products, the moulds which produce tham and the methods of detection, prevention and reduction of TMP contamination., Während einer langen Reifung wird die Oberfläche traditioneller Dauerfleischprodukte (TMP) normalerweise durch Schimmelpilze befallen. Unter ihnen produzieren toxikogenetische Spezies Mykotoxine als Sekundärmetabolite, die sich sehr nachteilig auf die Gesundheit von Mensch und Tier auswirken. Aflatoxine und Ochratoxine haben im Körper nachweislich karzinogene und mutagene Wirkungen; ihr Auftreten in Dauerfleischprodukten ergibt sich am häufigsten aus einem übermäßigen Wachstum von Schimmelpilzen. Neben diesem direkten Weg können Mykotoxine in Dauerfleischprodukten auch indirekt durch kontaminierte Futtermittel vorhanden sein, deren Verwendung bei Nutztieren zu Verschleppungseffekten in Fleischerzeugnissen führt. Aufgrund der Häufigkeit der Kontamination sind die Entwicklung von Analyseverfahren zum Nachweis von Mykotoxinen und die Anwendung von Methoden zur Identifizierung von Oberflächenschimmel von großer Bedeutung. Um eine Fleischkontamination und mögliche nachteilige Auswirkungen auf die Gesundheit der Verbraucher zu verhindern, sollten alle vorbeugenden Maßnahmen ergriffen und eine systematische Kontrolle dieser Kontaminanten vorgenommen werden. Diese Arbeit gibt einen Überblick über die Eigenschaften von Aflatoxinen und Ochratoxinen, die als Mykotoxine von größter Bedeutung für das Auftreten in Fleischprodukten sind, die Schimmelpilze, die diese produzieren, sowie die Methoden zum Nachweis, zur Verhinderung und Verringerung der Kontamination von Dauerfleischprodukten., Nel corso del lungo processo di stagionatura, la superficie dei tradizionali prodotti secchi a base di carne (TPC) viene solitamente ricoperta di muffa. Tra esse, le specie tassicogene producono metaboliti secondari delle micotossine che hanno conseguenze estremamente nocive per la salute dell’uomo e degli animali. Le aflatossine e le ocratossine hanno comprovati effetti cancerogeni e mutageni nell’organismo, e la loro presenza nei TPC è il più delle volte conseguenza dell’eccessiva crescita delle muffe superficiali. Oltre che direttamente, le micotossine possono essere presenti nei TPC anche indirettamente, mediante il mangime contaminato assunto degli animali, il cui impiego in zootecnia comporta la trasmissione (effetto carry-over) nei prodotti finali a base di carne. A causa della frequenza delle contaminazioni, è importante lo sviluppo dei metodi analitici di conferma per la rilevazione delle micotossine e l’applicazione del metodo per l’identificazione delle muffe superficiali. Per prevenire la contaminazione dei TPC e i possibili effetti nocivi per la salute del consumatore, è necessario adottate tutte le misure preventive e eseguire un controllo sistematico di questi agenti contaminanti. Questo studio dà un quadro delle proprietà delle aflatossine e delle ocratossine quali micotossine di maggior rilievo per la loro presenza nei prodotti a base di carne, delle muffe che le producono e dei metodi di rilevazione, prevenzione e riduzione della contaminazione dei TPC., Durante la larga maduración, la superficie de los productos cárnicos tradicionales crudo-curados (PCT) normalmente está cubierto de molde. Entre ellos, los tipos de molde toxicogénicos producen las micotoxinas como metabolitos secundarios, los cuales tienen efectos sumamente adversos sobre la salud humana y animal. Las aflatoxinas y ocratoxinas tienen efectos carcinogénicos y mutagénicos en el organismo comprobados su manifestación en los PCT en general es la consecuencia del crecimiento excesivo del molde sobre la superficie. Además de esta manera directa, las micotoxinas pueden aparecer en PCT indirectamente a través del pienso contaminado usado para los animales de granja y a través del efecto residual (carry-over effect) en los productos cárnicos finales. Debido a la frecuencia de la contaminación es importante desarrollar los métodos analíticos confirmatorios para la detección de las micotoxinas, y la aplicación de los métodos de identificación de los moldes en la superficie. Para evitar la contaminación de los PCT y posibles efectos nocivos sobre la salud de los consumidores, es necesario tomar medidas de prevención y llevar a cabo un control sistemático de estos contaminantes. Este trabajo da un resumen de las propiedades de las aflatoxinas y ocratoxinas como tipos de micotoxinas de mayor importancia para la manifestación en los productos cárnicos, de los moldes que les producen y de los métodos de detección, de prevención y de reducción de la contaminación de los PCT.

Assessment of microbiological quality of honey is very important, because of its use as a supplement in the human diet. Fungal contamination of food poses a challenge to public health and global food security. Although honey is known for its antifungal activity, there are not many species of fungi that have been tested. In this study, 30 honey samples collected in various parts of Croatia during the period of 2012-2017 were tested for the presence of yeasts and moulds through mycological analysis. The mycological identification showed that out of nine genera recovered, Cladosporium was the most frequent genus, followed by Penicillium and Alternaria, being found in 47.37% (9 samples), 10.53% (2 samples) and 10.53% (2 samples). In addition, genera of Mucor, Aureobasidium, Acremonium, Botrytis, Stachybotrys and Paecylomyces were isolated in 5.26% samples. The results showed that most of the moulds identified in this study are commonly found in honey.

The aim of this study was to investigate the occurrence of citrinin in different cereals (n = 341) and feedstuffs (n = 67) coming from farms and feed factories established in Croatia and Bosnia & Herzegovina. The highest mean citrinin concentration (103 ± 102 µg/kg) was observed in cereals sampled in Bosnia & Herzegovina during 2014, although significantly different levels between the two countries were not observed. Across the cereal samples, the maximal citrinin concentration was determined in wheat (429 µg/kg), while across the feedstuffs, the highest concentration was found in pig feed (63 µg/kg). Despite of the increased citrinin levels found in some samples, especially wheat, the obtained values cannot be compared against the maximum limits, since no such levels are stipulated under the applicable legislation. But, given that data on citrinin are very scarce, they can serve as an indicator of cereal and feed contamination in this part of Europe.

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 &amp, 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 &lt, 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 &amp, fatty acid composition and, to a lesser extent, in salt content, however not at the expense of product safety, quality and acceptability.

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

Although many countries prohibit whaling, it remains a significant cause of whale population decline. The meal of whales, likely killed during illegal whaling or caught accidentally, can ap- pear in a fish meal as a contaminant detectable by microscopic examination of bone fragments. In order to provide a rigorous basis for such a detection, microscopic characterization of bone fragments of 10 female and 10 male, less than a year to 21 years old bottlenose dolphins (Tur- siops truncatus), a widespread cetacean species herein used as a whale model, was performed and compared to the reference bone fragments of fish, ruminant, poultry and pig. The processing of bones mimicked that used in the production of meat and bone meals, while their description was based on qualitative characteristics, i.e. the shape of a bone fragment ; the shape, density and distribution of osteocyte lacunae, and the distribution and density of canaliculae. Bottlenose dol-phin bone fragments are smoothly contoured ; their elliptical osteocyte lacunae are clearly visible, while the canaliculae radiate from the lacunae in all directions. The comparison of these qualita-tive bone characteristics with that of other vertebrata under study revealed that bottlenose dol-phin bone fragments can definitely be differentiated from that of fish, may be differentiated from that of poultry and pig, but cannot be differentiated from that of ruminants. Measurements of the osteocyte lacunae showed the lacunar length to be strongly associated with animal’s age and lacunar shape, while their width was strongly associated with animal’s gender lacunar shape. The results indicate the possibility of detecting a whale meal admixture in a fish meal using light microscopy, which should be followed by PCR to enable the identification of the admixture source.

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

Cilj rada bio je ispitati kemijski sastav i pojavnost mikotoksina u tradicionalnim mesnim proizvodima (TMP) podrijetlom s domacinstava Bosne i Hercegovine. U razdoblju od 2015. do 2016., ukupno je uzorkovano 55 tradicionalnih mesnih proizvoda, od toga 25 kobasica proizvedenih od svinjskog mesa, 15 kobasica proizvedenih od mjesavine svinjskog i goveđeg mesa, 7 uzoraka pancete i 8 uzoraka prsuta. Osnovni kemijski i masno-kiselinski sastav određen je primjenom ISO metoda, dok su razine aflatoksina B1 (AFB1) i okratoksina A (OTA) određene primjenom imunoenzimskih ELISA metoda. Rezultati su pokazali varijacije u vrijednostima kemijskih parametara između TMP-a, s znacajno vecim (p < 0, 05) udjelom stearinske kiseline, te manjim udjelom linolne kiseline u kobasicama proizvedenim od mjesavine svinjskog i goveđeg mesa, u odnosu na TMP proizvedene u cijelosti od svinjetine. OTA je određen u 7 uzoraka razlicitih vrste proizvoda s najvecom koncentracijom od 6, 20 μg/kg, dok je AFB1 određen samo u jednoj kobasici od svinjskog mesa s koncentracijom neznatno vecom od limita kvantifikacije metode (1, 91 μg/kg). Utvrđena kontaminacija TMP-a mikotoksinima, posebno sa OTA, pokazuje da bi u cilju sprjecavanja kontaminacije meso i mesni proizvodi iz domacinstava trebali biti proizvedeni pod standardiziranim i dobro kontroliranim uvjetima.

Mycotoxins may contaminate food of animal origin due to the carry-over effect and represent a potential risk to human health. The problem of Fusarium mycotoxin contamination becomes an issue especially during rainy years characterised by substantial temperature changes. The aim of this study was to investigate into the level of Fusarium mycotoxins zearalenone (ZEN), deoxynivalenol (DON) and fumonisins (FUM) in maize silage (n=21), concentrated dairy cattle feeds (n=56) and cow milk samples (n=105), taken during 2015 from households located in four Croatian regions. The presence of mycotoxins was determined using validated ELISA methods. A high level of feedstuffs’ contamination was evidenced, especially with ZEN, with values higher than recommended observed in 9.5 % of maize silage samples. Fourteen point three percent (14.3 %) of milk samples were DON positive, with the toxin concentrations ranging from 5.4 to 67.3 μg/L. ZEN was determined in 94.3 % of milk samples, ranging from 0.3 to 88.6 μg/L. FUM were not detected in any of the analysed milk samples. Given the tolerable daily intakes (TDIs) defined for these mycotoxins, human health risks arising from the consumption of cow milk can generally be considered low, even in times characterised by weather conditions that facilitate the production of Fusarium mycotoxins in cereals subsequently used as dairy cattle feed. The exception represents particular milk samples in which high ZEN concentrations were found., Mikotoksini mogu kontaminirati hranu životinjskog podrijetla putem carry-over efekta i predstavljaju potencijalni rizik za ljudsko zdravlje. Problem kontaminacije mikotoksinima iz roda Fusarium izražen je naročito tijekom kišovitih godina, koje ujedno karakteriziraju i značajne promjene temperature. Cilj ovog istraživanja bio je ispitati razinu fuzarijskih mikotoksina zearalenona (ZEN), deoksinivalenola (DON) i fumonizina (FUM) u kukuruznoj silaži (n=21), koncentriranoj hrani za mliječne krave (n=56) i uzorcima kravljeg mlijeka (n=105), uzorkovanih tijekom 2015. godine sa obiteljskih poljoprivrednih gospodarstava iz četiri hrvatske regije. Prisutnost mikotoksina određena je primjenom ELISA validiranih metoda. Utvrđena je visoka razina kontaminacije stočne hrane posebno za ZEN, s vrijednostima većim od preporučenih u 9,5 % uzoraka kukuruzne silaže. Na DON je bilo pozitivno 14,3 % uzoraka mlijeka, s koncentracijom u rasponu od 5,4 do 67,3 μg/L. ZEN je određen u 94,3 % uzoraka mlijeka, u rasponu od 0,3 do 88,6 μg/L. FUM nije određen u niti jednom od analiziranih uzoraka mlijeka. S obzirom na prihvatljiv dnevni unos (TDI) definiran za ove mikotoksine, zdravstveni rizik za ljude koji proizlazi iz konzumacije kravljeg mlijeka općenito se može smatrati niskim, čak i tijekom razdoblja koja karakteriziraju vremenski uvjeti pogodni za proizvodnju fuzarijskih mikotoksina u žitaricama, a koje se nadalje koriste kao hrana za muzne krave. Izuzetak predstavljaju pojedini uzorci mlijeka u kojima su određene visoke koncentracije ZEN.

Cilj rada bio je ispitati kemijski sastav i pojavnost mikotoksina u tradicionalnim mesnim proizvodima (TMP) podrijetlom s domaćinstava Bosne i Hercegovine. U razdoblju od 2015. do 2016., ukupno je uzorkovano 55 tradicionalnih mesnih proizvoda, od toga 25 kobasica proizvedenih od svinjskog mesa, 15 kobasica proizvedenih od mješavine svinjskog i goveđeg mesa, 7 uzoraka pancete i 8 uzoraka pršuta. Osnovni kemijski i masno-kiselinski sastav određen je primjenom ISO metoda, dok su razine aflatoksina B1 (AFB1) i okratoksina A (OTA) određene primjenom imunoenzimskih ELISA metoda. Rezultati su pokazali varijacije u vrijednostima kemijskih parametara između TMP-a, s značajno većim (p < 0,05) udjelom stearinske kiseline, te manjim udjelom linolne kiseline u kobasicama proizvedenim od mješavine svinjskog i goveđeg mesa, u odnosu na TMP proizvedene u cijelosti od svinjetine. OTA je određen u 7 uzoraka različitih vrste proizvoda s najvećom koncentracijom od 6,20 µg/kg, dok je AFB1 određen samo u jednoj kobasici od svinjskog mesa s koncentracijom neznatno većom od limita kvantifikacije metode (1,91 µg/kg). Utvrđena kontaminacija TMP-a mikotoksinima, posebno sa OTA, pokazuje da bi u cilju sprječavanja kontaminacije meso i mesni proizvodi iz domaćinstava trebali biti proizvedeni pod standardiziranim i dobro kontroliranim uvjetima., Ziel dieser Arbeit bestand in der Untersuchung der chemischen Zusammensetzung und der Anwesenheit von Schimmelpilzgiften in traditionellen Fleischprodukten, die von Bauernhöfen aus Bosnien und Herzegowina stammen. Im Zeitraum von 2015 bis 2016 wurden insgesamt 55 Stichproben von traditionellen Fleischprodukten genommen, davon 25 Schweinefleischwürste, 15 Würste aus gemischtem Schweine- und Rindfleisch, 7 Speckproben und 8 Proben von luftgetrocknetem Rohschinken. Die grundlegende chemische Zusammensetzung und der Anteil an Fettsäuren wurde anhand von ISO-Methoden ermittelt, während der Gehalt an Aflatoxin B1 (AFB1) und Ochratoxin A (OTA) durch die Anwendung von antikörperbasierten Nachweisverfahren (ELISA) ermittelt wurde. Die Ergebnisse wiesen Variationen in Bezug auf die Werte chemischer Parameter bei traditionellen Fleischprodukten auf, mit einem bedeutend höheren (p < 0,05) Anteil der Stearinsäure und einem niedrigeren Anteil der Linolsäure in Würsten, die aus einer Mischung von Schweine- und Rindfleisch hergestellt wurden im Vergleich zu reinen Schweinefleischprodukten. Ochratoxin A wurde in 7 Proben unterschiedlicher Produkte mit einer maximalen Konzentration von 6,20 µg/kg ermittelt, während AFB1 nur in einer Schweinefleischwurst mit einer Konzentration kaum über dem Limit der Quantifizierungsmethode (1,91 µg/kg) festgestellt wurde. Die festgestellte Kontaminierung von traditionellen Fleischprodukten durch Schimmelpilzgifte, insbesondere OTA, weist darauf hin, dass zur Vorbeugung der Kontaminierung das Fleisch und Fleischprodukte auf den Bauernhöfen unter standardisierten und streng kontrollierten Bedingungen hergestellt werden sollten., Lo studio aveva lo scopo di analizzare la composizione chimica e la presenza delle micotossine nei prodotti tradizionali a base di carne (PTC) prodotti in ambito domestico nella Bosnia e Erzegovina. Tra il 2015 e il 2016 è stato eseguito il campionamento di 55 prodotti tradizionali a base di carne, di cui 25 salsicce prodotte con carne suina, 15 salsicce prodotte con carne suina e bovina, 7 campioni di pancetta e 8 campioni di prosciutto crudo. La composizione chimica e degli acidi grassi è stata stabilita con l’applicazione del metodo ISO, mentre i livelli dell’aflatossina B1 (AFB1) e dell’ocratossina A (OTA) sono stati accertati mediante il test immunoenzimatico ELISA. I risultati hanno evidenziato variazioni nei valori dei parametri chimici tra PTC, con una presenza molto significativa (p < 0,05) di acido stearico ed una minore presenza dell’acido linoleico nelle salsicce prodotte con carne suina e bovina, rispetto ai PTC prodotti integralmente con carne suina. La presenza della OTA è stata accertata in 7 campioni di differenti tipi di prodotto, con una concentrazione massima di 6,20 µg/kg, mentre la presenza della AFB1 è stata riscontrata solo in un campione di salsiccia prodotta con carne suina, avente una concentrazione di pochissimo superiore al limite del metodo impiegato per la quantificazione (1,91 µg/kg). Accertata la presenza di micotossine nei PTC, in particolare delle OTA, si ritiene necessario che la carne e i prodotti a base di carne prodotti in ambito domestico, al fine di prevenire la contaminazione, siano prodotti in condizioni standardizzate e rigidamente controllate., El fin de este trabajo fue investigar la composición química y la manifestación de los micotoxinas en productos cárnicos tracidionales (PCT) de los hogares de Bosnia y Herzegovina. En el período entre 2015 y 2016 fueron tomadas 55 muestras de productos cárnicos tradicionales, entre cuales fueron 25 salchichas de carne de cerdo, 15 salchichas de la mezcla de carne de cerdo y carne bovina, 7 muestras de panceta y 8 muestras de jamón. La composición básica química y la composición de ácidos grasos fueron determinadas por el método ISO, mientras los niveles del aflatoxina (AFB1) y del ocratoxina A (OTA) fueron determinados por los métodos inmunoenzimáticos ELISA. Los resultrados mostraron fluctuaciones en los valores de los parámetros químicos entre los PCT, con la proporción del ácido esteárico significativamente más alta (p < 0,05) y con la poporción más baja del ácido linoléico en las salchichas hechas de la mezcla de carne de cerdo y carne bovina, con respecto a los PCT hechos enteramente de la carne de cerdo. El OTA fue determinado en 7 muestras de diferentes tipos de productos con la concentración más alta 6,20 µg/kg, mientras el AFB1 fue encontrado solamente en una salchicha de carne de cerdo, con ligera diferencia entre la concentración y el límite de cuantificación (1,91 µg/kg). La determinada contaminación de los PCT por los micotoxinas, especialmente por el OTA, muestra que la carne y los productos cárnicos de los hogares deberían ser producidos bajo condiciones estrandarizadas y bien controladas con el fin de prevenir la contaminación.