Occurrence and Health Risk Assessment of Cadmium Accumulation in Three Tricholoma Mushroom Species Collected from Wild Habitats of Central and Coastal Croatia.

This study deals with the biomonitoring of cadmium (Cd) heavy metal in the three selected Tricholoma mushroom species collected from wild habitats of central and coastal Croatia. For this, mushroom (T. columbetta: n = 38, T. portentosum: n = 35, and T. terreum: n = 34) and surface soil samples were collected from nine forest localities of Croatia and analyzed for Cd concentration using inductively coupled plasma−optical emission spectrometry (ICP−OES) through the acid digestion method. The findings revealed that Cd was present in Tricholoma spp. and surface soil. However, the maximum mean Cd concentration (mg/kg dry weight) was recorded in T. portentosum (cap: 0.98; stipe: 0.72), followed by T. columbetta (cap: 0.96; stipe: 0.73) and T. terreum (cap: 0.81; stipe: 0.63). The bioconcentration factor (BCF) value (>1) revealed that the selected Tricholoma spp. had the potential for Cd accumulation. Moreover, the principal component (PC) and hierarchical cluster (HC) analyses were used to derive the interactions and similarities between Cd levels Tricholoma spp. and sampling localities. The multivariate analysis suggested that central sampling localities had higher Cd levels as compared to coastal localities. However, the daily intake of metals (DIM < 0.426) and health risk index (HRI < 1) showed that there was no potential health risk associated with the consumption of selected Tricholoma spp. The findings of this study are helpful to understand the Cd accumulation behavior of wild edible Tricholoma spp. collected from Croatia.

The Occurrence of Five Unregulated Mycotoxins Most Important for Traditional Dry-Cured Meat Products.

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 B1 (AFB1), 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 AFB1 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.

Sustainable Use of Sewage Sludge as a Casing Material for Button Mushroom (Agaricus bisporus) Cultivation: Experimental and Prediction Modeling Studies for Uptake of Metal Elements.

The present study focused on the use of sewage sludge (SS) as a casing material amendment and the potential uptake of metal elements by the cultivated white button (Agaricus bisporus: MS-39) mushroom. Laboratory experiments were performed under controlled environmental conditions to grow A. bisporus on the composted wheat straw substrate for 50 days. Different treatments (0, 50, 100, 150, and 200 g/kg) of casing material were prepared by mixing garden and dried SS and applied on the mushroom substrate after proper sterilization. The results revealed that SS application was significant (p < 0.05) in accelerating mushroom yield with a biological efficiency of 65.02% for the mixing rate of 200 g/kg. Moreover, the maximum bioaccumulation of selected metal elements (Cu, Cr, Cd, Fe, Mn, and Zn) was observed using the same treatment. Additionally, the multiple regression models constructed for the uptake prediction of metal elements showed an acceptable coefficient of determination (R2 > 0.9900), high model efficiency (ME > 0.98), and low root mean square error (RMSE < 0.410) values, respectively. The findings of this study represent sustainable use of SS for the formulation of mushroom casing material contributing toward synergistic agro-economy generation and waste management.

Proteolytic Lactococcus lactis and Lipolytic Enterococcus durans of Dairy Origin as Meat Functional Starter Cultures.

RESEARCH BACKGROUND: As fermentation is an integral feature of both, dry sausage and cheese production, this has led to the evaluation of bacterial cultures Lactococcus lactis ssp. cremoris (LL8307) and Enterococcus durans (ED0207) originally isolated from artisanal Croatian hard type cheese to diversify the range of flavours of dry fermented sausages and to increase their microbiological safety. Both strains were chosen for their high or medium acidifying, proteolytic and/or lipolytic activity and bioprotective potential after step-by-step selection of wild isolates. Therefore, this study aims to evaluate the survival rate of selected starter cultures in wild boar meat sausages during the ripening period of 40 days at a local small-scale facility under artisanal conditions as well as their influence on sausage quality parameters. EXPERIMENTAL APPROACH: Safety, biotechnological and probiotic properties of twenty-three enterococcal and lactococcal isolates of dairy origin were studied. Based on the results, two best candidates were selected and added to the meat batter during the artisanal wild boar meat sausage preparation where their survival rate, effect on physicochemical, microbiological and sensorial properties and histamine content were evaluated. RESULTS AND CONCLUSIONS: As revealed by repetitive element-polymerase chain reaction (rep-PCR), native starter cultures survived up to 15 days of ripening and were either absent from (LL8307) or reduced by 80% (ED0207) in final products. The application of native starter cultures rapidly decreased pH (p<0.05), leading to the significantly lower load of E. coli, coliforms and Enterobacteriaceae in ready-to-eat sausages prepared by the addition of starter cultures (3.04-3.94 log CFU/g) than in the control (3.88-5.00 log CFU/g). Analysis of hedonic test data revealed that some of the sensory traits (odour, flavour, juiciness) of treatments with starter cultures were highly liked by the higher percentage of consumers. The results suggest that these starter cultures would represent a valuable tool to improve the homogeneity of artisanal manufacture and hygienic quality of fermented sausages and can be safely used for food application. NOVELTY AND CCIENTIFIC CONTRIBUTION: This is the first study to explore in depth the biotechnological potential of bacterial cultures isolated from artisanal Croatian cheese as functional starter cultures for high-quality game meat sausage production.

Plant Carotenoids as Pigment Sources in Laying Hen Diets: Effect on Yolk Color, Carotenoid Content, Oxidative Stability and Sensory Properties of Eggs.

The aim of this study was to evaluate the effect of a supplementation diet for hens consisting of dried basil herb and flowers of calendula and dandelion for color, carotenoid content, iron-induced oxidative stability, and sensory properties of egg yolk compared with commercial pigment (control) and marigold flower. The plant parts were supplemented in diets at two levels: 1% and 3%. In response to dietary content, yolks from all diets differed in carotenoid profile (p < 0.001). The 3% supplementation level resulted in a similar total carotenoid content as the control (21.25 vs. 21.79 µg/g), but by 3-fold lower compared to the 3% marigold (66.95 µg/g). The tested plants did not achieve yolk color fan values as the control (13.47) or 3% marigold (11.47), and among them, calendula had the highest values (9.73). Despite the low carotenoid content in diets supplemented with basil herb, iron-induced malondialdehyde (MDA) concentration was low as for marigold (on average 106.83 vs. 92.68 ng/g after 250 min). The treatments differed in sensory color scores for fresh and hard-boiled yolks and flavor while other sensory properties were similar. In conclusion, the supplementation of plants in a hen diet may result in yolks containing carotenoids and other compounds showing a high antioxidant effect.

The survival rate and efficiency of non-encapsulated and encapsulated native starter cultures to improve the quality of artisanal game meat sausages.

This study addresses the application of native, multiple strain starter cultures for standardization of game meat sausages production. The designed starter cultures consisting of two indigenous Lactobacillus sakei and one Leuconostoc mesenteroides strains. These strains were used in both, the encapsulated and non-encapsulated form, in the game meat dough, individually or in combination, with eight treatments in total. Microbiological and physicochemical characteristics of the sausages were monitored throughout the manufacturing process, while sensory properties, biogenic amine content, and volatile compounds were evaluated in the final products. As revealed by rep-PCR, native starter cultures, encapsulated or non-encapsulated, had survived the whole sausage production process; however, to varying degrees. The application of indigenous decarboxylase negative Lb. sakei strains significantly (P < 0.05) reduced tyramine content, rapidly decreased pH and promoted the number reduction of Enterobacteriaceae and elimination of E. coli, L. monocytogenes and coliforms in ready-to-eat products. A total of 84 volatile compounds were identified by SPME-GC-MS in the eight treatment batches of game meat sausages, with only minor differences between the treatments. No significant differences in sensory traits (P > 0.05) between tested treatments were found, although treatment with the Lb. sakei strains received the highest scores for the sensory traits including cross-section, odour, hardness, aroma, and overall acceptability. Combination of multi-strain Lb. sakei starter cultures resulted in growth prevention of undesirable microbiota, reduction of tyramine content and increased the acceptability parameters of full-ripened sausages, which make them good candidates for industrial as well as artisanal application.

Effect of Different Method of Drying of Five Varieties Grapes (Vitis vinifera, L.) on the Bunch Stem on Physicochemical, Microbiological, and Sensory Quality.

The influence of the drying technique on certain quality attributes of dried grapes was analysed. Five varieties of grape were used in this study (Bezsemenné, Perlette, Vrboska, Beauty seedless, and Jakubské). All the varieties were dried using four methods: drying at 40 °C, combined drying (70, 65, and 60 °C), drying at 70 °C, and lyophilisation. The quality attributes such as total soluble solids, water activity, content of vitamin C, the colour parameters (L*, a*, b*), and microbiological (colony forming units, moulds, yeasts) and sensory (smoothness, gloss, colour, odour, chewability, juiciness, flavour, and overall impression) analysis were determined, depending on the method of drying. Lyophilised grapes from the Jakubské variety had the highest vitamin C content. They contained 0.58 mg of vitamin C per 100 g of dried grapes. However, no significant differences between varieties in vitamin C content were found. This confirmed concerns about the negative impact on the nutritional quality of dried grapes with increasing temperature. The lowest total count of microorganisms (CFU), moulds, and yeasts were determined after using highest drying temperature (70 °C). Higher CFU values were determined in lyophilised grapes, and then followed by grapes dried at 40 °C. Lyophilised grapes and grapes dried at 40 °C achieved the best sensory rating in general. The final product was intended to have a unique appearance while maintaining the evaluated quality parameters.

Characterization of Traditional Croatian Household-Produced Dry-Fermented Sausages.

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.

Analysis of phthalic acid esters in agricultural soils.

The aim of the study was monitoring of phthalic acid esters in agricultural soils of the Czech Republic over the period of 6 years, namely dibutyl phthalate (DBP) and di-2-ethylhexyl phthalate (DEHP). Monitoring took place in twelve regions of the Czech Republic. Soil samples were taken evenly from the top and bottom soil horizons using a zig-zag pattern. Soil samples were taken from arable land, permanent grassland, and hop field. Lyophilisation of the samples was done by ultrasound-assisted extraction using a mixture acetone-hexane (1:1). Phthalic acid esters were analysed using high-performance liquid chromatography with UV detection. Subsequently, the results were statistically compared by analysing the principal components (PCA) to determine the effect of individual factors on the content of phthalic acid esters in agricultural soil. Factors such as precipitation, distance from a pollution source, amount of pesticides, and amount of artificial and organic fertilizers were taken into account. If we compare the concentrations established in this study with the limits set out in the Methodological Instruction of the Ministry of the Environment of the Czech Republic based on RSLs (Regional Screening Levels) issued by the USEPA (United States Environmental Protection Agency), none of these values were exceeded.

The Influence of Meat Batter Composition and SausageDiameter on Microbiota and Sensory Traits of ArtisanalWild Boar Meat Sausages.

In this study, the influence of meat batter composition and sausage diameter on the development of microbiota and sensory traits of traditional, spontaneously fermented wild boar meat sausages are evaluated. This research also demonstrates how principal component analysis (PCA) can be used to relate product sensory properties to particular microbial genotype and to select potential starter or adjunct culture. Generally, similar microbiological results were obtained in all types of products. The undesirable microbiota was either not detected at any sausage production stage or its number decreased below the detection limit in ripened sausages. The low growth rate of lactic acid bacteria (LAB) was consistent with the obtained pH and slow acidification rate. Although no differences in the composition of LAB species were noticed between sausage types (50S=50% wild boar meat in small casing, 50L=50% wild boar meat in large casing, 100S=100% wild boar meat in small casing), a clear separation based on LAB genotypes could be observed. Upon quantitative descriptive analysis, significant differences in sensory attributes between sausage types were established. According to the PCA, the overall acceptability traits of sausages are closely linked to one Leuconostoc mesenteroides genotype (LM_4). Of all tested technological properties, LM_4 strains showed remarkable acidification ability, lowering the pH from pH=5.41 to 3.74, and pronounced proteolytic activity on skimmed milk as well as antagonistic activity against Staphylococcus aureus (DSM 20231) and Brochothrix thermosphacta (LMG 17208). Lipolytic and haemolytic activities were not detected, and all analyzed strains were susceptible to tested antibiotics and possessed no biogenic amine genes.

Heavy metal bioaccumulation by wild edible saprophytic and ectomycorrhizal mushrooms.

Heavy metals cause serious problems in the environment, and they can be accumulated in organisms, especially in the higher fungi. The concentration of Ni, Cr, Pb, Cd, and Hg in 10 species of edible mushrooms in Medvednica Nature Park, Croatia was therefore determined. In addition, the similarity between the studied species was determined by cluster analysis based on concentrations of the aforementioned metals in the fruiting bodies. The contents of nickel, chromium, lead, cadmium, and mercury in the fruiting bodies of mushrooms were obtained by X-ray fluorescence spectrometry. The highest concentrations of Ni (3.62 mg kg(-1)), Cr (3.01 mg kg(-1)), and Cd (2.67 mg kg(-1)) were determined in Agaricus campestris. The highest concentration of Pb (1.67 mg kg(-1)) was determined in Macrolepiota procera, and the highest concentration of Hg (2.39 mg kg(-1)) was determined in Boletus edulis. The concentration of all heavy metals significantly differed (p < 0.001) between examined saprophytic and ectomycorrhizal mushrooms. Considering anatomical part of the fruiting body (cap-stipe), a considerably higher concentration of the analyzed elements was found in the cap for all mushroom species. According to calculated bioconcentration factors, all the examined species were found to be bioexclusors of Ni, Cr, and Pb and bioaccumulators of Cd and Hg. Cluster analysis performed on the basis of the accumulation of the studied metals revealed great phenotypic similarity of mushroom species belonging to the same genus and partial similarity of species of the same ecological affiliation.