Chemical composition and evaluation of antioxidant, antimicrobial and antiproliferative activities of Tuber and Terfezia truffles.

Ten truffle species of Tuber and Terfezia genera were chemical characterized, assessing their proximate composition, individual nutrient compounds and some bioactive molecules. The bioactive properties of these species were also evaluated, namely their antioxidant, antimicrobial and cytotoxic potential. Carbohydrates were the main macronutrients present in truffles, followed by proteins. Furthermore, the levels of polyunsaturated fatty acids (PUFA), subsequently presented as a percentage, were higher in truffles (38.2-79.3%) except in Tuber magnatum and Terfezia arenaria, which have a more saturated fatty acids (SFA) profile (70.7% and 53.7%, respectively). Comparing the species, T. magnatum revealed the highest levels of total phenolic compounds (TPC) (290 mg GAE/100 g truffle), as also the best results in the four methods used to evaluate the antioxidant activity. On the other hand, only five extracts obtained from some studied truffle species (Terfezia magnusii, Tuber aestivum, Tuber gennadii, and Tuber melanosporum) showed a slight inhibition of microbial growth, tested against different bacteria. Terfezia and T. gennadii extracts, showed potential to inhibit the cellular growth of NCI-H460, HeLa, HepG2, and MCF-7 cell lines (GI50 concentrations range: 19-78, 33-301, 83-321 and 102-321 µg/mL, respectively), indicating anti-proliferative activity. Nevertheless, T. arenaria revealed some potential hepatotoxicity, inhibiting the growth of PLP2 cells (GI50 concentration of 220 µg/mL), a primary cell culture obtained from porcine liver.

Effects of gamma irradiation on the shelf-life and bioactive compounds of Tuber aestivum truffles packaged in passive modified atmosphere.

The effects of gamma irradiation (0.5, 1.0, 1.5 and 2.5 kGy doses) on Tuber aestivum packaged under modified atmosphere was evaluated. The respiration rate, microbial populations, sensory characteristics and content of bioactive compounds (total carbohydrates, chitins, ß-glucans, proteins, total phenols and sterols) were monitored from immediately after treatment up to day 42 of storage at 4 °C. All the irradiation treatments tested reduced the microbial groups studied by more than 3 log cfu/g. Increasing irradiation doses slowed down the subsequent microbial development throughout the conservation period for all the groups studied. The irradiation treatments did not negatively affect truffle sensory characteristics. Only a slight visible superficial yeast growth was detected at the end of the shelf-life in all doses applied. Total carbohydrate content, chitins, ß-glucans and proteins levels were not affected after irradiation. However, sterols, particularly stigmasterol, slightly decreased after irradiation, while levels of phenolic compounds doubled during storage. Gamma irradiation (2.5 kGy) could be used to extend the shelf-life of summer truffles packaged under modified atmosphere, since no remarkable reduction of bioactive compounds were noticed after 42 days of storage, and their sensory and microbial parameters were of higher quality than those of non-irradiated controls.

Effects of electron-beam and gamma irradiation treatments on the microbial populations, respiratory activity and sensory characteristics of Tuber melanosporum truffles packaged under modified atmospheres.

The effects of electron-beam or gamma irradiation (doses of 1.5 kGy and 2.5 kGy of either one) on the microbial populations, respiratory activity and sensory characteristics of Tuber melanosporum packaged under modified atmospheres were monitored immediately after treatment, and subsequently every seven days during 35 days of storage at 4 °C. Treatments with 1.5 and 2.5 kGy reduced the total mesophilic aerobes counts respectively by 4.3 and 5.6 log cfu/g for electron-beam treatment, and by 6.4 and 6.6 log cfu/g for gamma irradiation. Other microbial groups studied (Pseudomonas genus, Enterobacteriaceae family, lactic acid bacteria, mesophilic aerobic spores, molds and yeasts) were not detected after the treatments. A decrease in the respiratory activity was detected in all the irradiated batches, indicating that the carbon dioxide levels were lower and the oxygen levels higher than those of the non-irradiated ones. Two species of yeasts, Candida sake and Candida membranifaciens var. santamariae, survived the irradiation treatments and became the dominant microbial populations with counts of up to 7.0 log cfu/g. The growth of these microorganisms was visible on the surface of irradiated truffles from day 21 onwards, affecting the flavor and the general acceptability of the ascocarps. Moreover, a watery exudate was detected in the treated truffles from the third week onwards, so the application of irradiation treatments in doses equal to or above 1.5 kGy did not preserve the quality characteristics of T. melanosporum truffles beyond 28 days.

Effects of electron-beam irradiation on the shelf life, microbial populations and sensory characteristics of summer truffles (Tuber aestivum) packaged under modified atmospheres.

The effects of two doses of electron-beam irradiation (1.5 kGy and 2.5 kGy) on the microbial populations (total mesophilic aerobes, Pseudomonas genus, Enterobacteriaceae family, molds and yeasts) and sensory characteristics of Tuber aestivum packaged under modified atmospheres were monitored immediately after treatment, and weekly during 42 days of storage at 4 °C. Treatment with 1.5 and 2.5 kGy reduced the pseudomonads populations by 4.3 and 5.5 logs, respectively. Enterobacteriaceae counts decreased by 5.4 logs with the 1.5 kGy dose and counts below the detection limit (<1.0 log cfu/g) were obtained with the 2.5 kGy dose. Lactic acid bacteria and yeasts were less affected by the ionizing radiation treatments and they became the dominant microbial populations throughout storage with microbial counts up to 7.1 log cfu/g. The carbon dioxide levels inside the packages containing irradiated truffles were lower than those of the non-irradiated ones, suggesting a decrease in the respiration rate of the treated ascocarps. The treatments with 1.5 and 2.5 kGy e-beam did not negatively affect the sensory characteristics of truffles, but a visible superficial yeast growth was detected in truffles irradiated with 1.5 kGy at the end of their shelf life (day 28). Treatment with 2.5 kGy e-beam has prolonged the shelf life to 42 days, compared with 21 days for the untreated samples.

Shelf-life extension of fresh Tuber aestivum and Tuber melanosporum truffles by modified atmosphere packaging with microperforated films.

The aim of this study was to design a modified atmosphere packaging suitable for Tuber melanosporum and Tuber aestivum truffles that extend their shelf life and their availability as a fresh product. Their respiration rates were determined by O(2) depletion and CO(2) formation in closed systems performed at different temperatures: 4, 10, and 23 degrees C. The results were fitted by exponential equations and derivatives of these equations were used to obtain the experimental respiration rates. Our results revealed high respiration rates in both species of truffles and respiratory quotients (RQ) higher than 1 in all the cases studied. A linear dependence of respiration rate, both R(O2) and R(CO2), on O(2) concentration was revealed. A mathematical model was used to predict the evolution of the gaseous composition at 4 degrees C in the interior of polypropylene trays (250 mL) heat sealed with 4 microperforated films of different transmission rates. A microperforated film with 2 holes (90 x 50 microm) was selected to produce an internal atmosphere of 15%CO(2)/7%O(2) at 4 degrees C. The predicted atmosphere composition was confirmed by the experimental results. The quality and microbiological characteristics of fresh truffles, packaged in these conditions, revealed that the microbial counts of pseudomonads and Enterobacteriaceae were decreased, the weight loss was reduced, the typical hard texture was maintained, and the development of mycelium growth was delayed, enabling good scores for aroma and flavor, and therefore prolonging the shelf life of T. melanosporum and T. aestivum truffles to 28 and 21 d, respectively. Practical Application: This study describes the benefits of using MAP with microperforated films in the postharvest storage of Tuber melanosporum and Tuber aestivum fresh truffles. The shelf life of T. aestivum is prolonged to 21 d and of T. melanosporum to beyond 28 d increasing the possibilities for a foreign market.

Diversity of culturable microorganisms and occurrence of Listeria monocytogenes and Salmonella spp. in Tuber aestivum and Tuber melanosporum ascocarps.

The aim of this study was to investigate the total mesophilic microorganisms, Pseudomonas genus, Enterobacteriaceae family, mold and yeast counts and the presence of Listeria monocytogenes and Salmonella spp on Tuber aestivum and Tuber melanosporum ascocarps. The results confirmed that the major percentage of the microorganisms, approximately 9.0 log ufc/g, were present in the peridium, the glebas of healthy truffles being practically free of microorganisms. The predominant microbial group was the Pseudomonas averaging 8.3 and 8.4 log cfu/g on T. aestivum and T. melanosporum whole ascocarps, respectively. The Enterobacteriaceae also achieved high populations, especially in T. aestivum truffles, with 6.3 log cfu/g. Molds and yeasts never exceeded 5.0 log cfu/g. The characterization of the isolates revealed that the fluorescens pseudomonads were the most prevalent. Raoultella terrigena and Enterobacter intermedius were the dominant Enterobacteriaceae. The identification of the yeast isolates revealed five species: Debaryomyces hansenii, Issatchenkia scutulata, Rhodotorula aurantiaca, Saccharomyces dairensis and Trichosporon beigelii subspecies A and B. The mold genera detected in both species of truffles were Aspergillus, Cladosporium, Penicillium and Fusarium, Trichoderma being present only in T. aestivum. L. monocytogenes was found in 10% of the samples of T. aestivum analysed but Salmonella spp. was not detected. Knowledge of the microbial population in terms of possible food borne and pathogen microorganisms is very useful for establishing successful disinfection and storage methods to prolong the shelf-life of ascocarps of T. aestivum and T. melanosporum.