BACKGROUND:Along bacteria, yeasts are common in forages and forage fermentations as spoilage microbes or as additives, yet few studies exist with species-level data on these fungi's occurrence in feedstuff. Active dry yeast and other yeast-based products are also common feed additives in animal husbandry. Here, we aimed to characterize both fermented and non-fermented milking cow feedstuff samples from Hungary to assess their microbial diversity in the first such study from Central Europe. RESULTS:We applied long-read bacterial metabarcoding to 10 fermented and 25 non-fermented types of samples to assess bacterial communities and their characteristics, surveyed culturable mold and yeast abundance, and identified culturable yeast species. Fermented forages showed the abundance of Aerococcaceae, Bacillaceae, Brucellaceae, Lactobacillaceae, Staphylococcaceae, and Thermoactinomycetaceae, non-fermented ones had Cyanothecaceae, Enterobacteriaceae, Erwiniaceae, Gomontiellaceae, Oxalobacteraceae, Rhodobiaceae, Rickettsiaceae, and Staphylococcaceae. Abundances of bacterial families showed mostly weak correlation with yeast CFU numbers, only Microcoleaceae (positive) and Enterococcaceae and Alcaligenaceae (negative correlation) showed moderate correlation. We identified 14 yeast species, most commonly Diutina rugosa, Pichia fermentans, P. kudriavzevii, and Wickerhahomyces anomalus. We recorded S. cerevisiae isolates only from animal feed mixes with added active dry yeast, while the species was completely absent from fermented forages. The S. cerevisiae isolates showed high genetic uniformity. CONCLUSION:Our results show that both fermented and non-fermented forages harbor diverse bacterial microbiota, with higher alpha diversity in the latter. The bacterial microbiome had an overall weak correlation with yeast abundance, but yeasts were present in the majority of the samples, including four new records for forages as a habitat for yeasts. Yeasts in forages mostly represented common species including opportunistic pathogens, along with a single strain of Saccharomyces used as a feed mix additive.

Pleurotus ostreatus (Jacq. ex Fr.) P. Kumm mushrooms are cultivated on diverse by-products based on substrates that hold promise for mitigating antibiotic usage in the poultry industry and reducing environmental pollution. By incorporating agricultural by-products into mushroom cultivation, the functionality of the mushroom products can be increased, then the final product can be a more effective feed supplement. After mushroom cultivation, spent mushroom substrate (SMS) can be valorized, due to the presence of huge amounts of bioactive compounds like β-glucan, chitin, polyphenols, and flavonoids related to mycelia. As a prebiotic and antimicrobial feed supplement, these mushrooms positively influence gut microbiota, intestinal morphology, and thus overall poultry well-being. This article underscores the potential of solid-state fermentation (SSF) to enhance the bioactivity of oyster mushrooms and their derivatives, offering a cost-effective and efficient strategy for transforming unconventional feeding materials. Moreover, it emphasizes broader implications, including the reduction of antibiotic dependence in poultry farming, highlighting the promising integration of oyster mushrooms and their derivatives for sustainable and environmentally conscious poultry production.

Oyster mushrooms (Pleurotus ostreatus L.) are renowned for their antioxidant, antimicrobial, and prebiotic properties. This study explores the antioxidant characteristics, activity, and β-glucan content in freeze-dried mushroom samples, investigating the influence of sous-vide cooking. Uncooked freeze-dried P. ostreatus and three pre-cooked freeze-dried samples (70, 80, 90 °C through 4 hours) were analysed for Total Polyphenol Content (TPC), Total Flavonoid Content (TFC), Radical Scavenging (DPPH), Ferric Reducing Antioxidant Power (FRAP), and β-glucans content via HPLC and Total Dietary Fiber (TDF) via enzymatic gravimetric method. Results indicate that uncooked mushroom powder exhibited superior antioxidant capabilities compared to cooked samples. The sous-vide cooked (80 °C) mushrooms displayed the highest total phenolic and flavonoid content. Moreover, pre-cooked (70 °C) mushroom powder demonstrated the highest β-glucan content, significantly surpassing the uncooked control sample. Notably, pre-cooked groups (80, 90 °C) demonstrated significantly higher TDF levels compared to uncooked sample. This research offers valuable insights into the potential use of mushrooms as high-antioxidant, antimicrobial, and prebiotic food or feed supplements, with broad implications across various fields.

This study aimed to examine the impact of mushroom extract-based solid media on the β-glucan content, growth rate, density, and biomass content of Pleurotus ostreatus (oyster mushroom) mycelia. Fresh, high-quality raw P. ostreatus were washed, sliced, and heated in a sealed pressure cooker at 90°C for 4 h in the drying cabinet. Following the heating process, centrifugation was carried out. Different concentrations of Pleurotus ostreatus extract were mixed with distilled water (0%, 25%, 50%, 75%, and 100%) and prepared for a sterile solid media. A malt extract-based medium was maintained as a control. This study focuses on the growth performance of P. ostreatus mycelium on its own mushroom extract-based culture medium which holds considerable economic and environmental significance. During the six-day observation period, the mycelium exhibited consistent growth across all tested media, maintaining a steady growth rate of 15 mm. The increased content of mushroom extract resulted from the enhanced density of the mycelia and biomass content. It can be inferred that when media containing less than 25% of mushroom extract dilution is used, β-glucan can be formed in smaller amounts. Further research is needed to explore mushroom extract derived from different types of mushroom byproducts, which fail to meet commercial standards.

Sour cherry is a non-climacteric fruit that quickly loses its quality after harvest, so effective storage and packaging are essential to minimize postharvest decay. Our study aimed to (i) evaluate fruit decay incidence during shelf-life for both freshly harvested and six-week-cold-stored sour cherry fruits, comparing normal atmospheric conditions with modified atmosphere packaging across three sour cherry cultivars (‘Érdi bőtermő’, ‘Újfehértói fürtös’, and ‘Petri’); and (ii) assess postharvest fruit decay incidence across the following three production systems: conventional, integrated pest management (IPM), and reduced IPM, for the cultivar ‘Érdi bőtermő’. The results showed that modified atmosphere packaging effectively preserved or slightly increased fruit firmness (ranging from 27.1 to 46%) compared to control fruits across all cultivars. Fruit weight loss (ranging from 1.18 to 26.1%) was also significantly reduced under modified atmosphere packaging compared to normal atmospheric storage. Over a 14-day period, the decay incidence of harvested fruits consistently increased, with major losses from day 6 onward (ranging from 17.7 to 77.0% at shelf-life day 14) across all cultivars and years. After six weeks of normal atmospheric cold storage, decay incidence remained low (ranging from 0 to 9.4%) and was comparable across all cultivars. Both normal atmospheric and modified atmosphere packaging storage maintained sour cherry quality during cold storage, but the shelf-life of the fruits was significantly shorter at room temperature, regardless of the storage methods. Fruit decay incidence at harvest was highest in the reduced IPM system (ranging from 3.2 to 6.4%), significantly exceeding those observed in the conventional and IPM systems. In post-MAP cold storage, decay incidence followed the following order: conventional < IPM < reduced IPM, with significant differences observed only between the conventional and reduced IPM systems. In conclusion, our results suggest that sour cherries produced under conventional or IPM systems and stored in modified atmosphere packaging offer an effective postharvest strategy for preserving fruit quality.

Pleurotus ostreatus (oyster mushroom) holds excellent promise worldwide, bringing several opportunities and augmenting the tool sets used in the biotechnology field, the food industry, and medicine. Our study explores the antimicrobial and probiotic growth stimulation benefits of freeze-dried P. ostreatus powders (OMP-TF, oyster mushroom powder from the total fresh sample; OMP-CSR, oyster mushroom powder from the cooked solid residue; OMP-CL, oyster mushroom powder from the cooked liquid), focusing on their bioactive compounds and associated activities. Our research examined polysaccharide fractions—specifically total glucans and α- and β-glucans—alongside secondary metabolites, including polyphenols and flavonoids, from freeze-dried mushroom powders. Additionally, carbon nanodots (CNDs) were also characterized. The growth inhibition was tested against Escherichia coli and Staphylococcus epidermidis, while the capacity for stimulating probiotic growth was evaluated using Lactobacillus plantarum and Lactobacillus casei. Evidence indicates that OMP-CL and OMP-CSR exhibit significant antimicrobial properties against S. epidermidis Gram-positive bacteria. OMP-CL notably promoted the growth of L. casei. OMP-CL, containing the most significant number of CNDs, has shown to be a valuable source for gut microbiota modulation, with its antimicrobial and probiotic-stimulating efficacy. However, further in vitro and in vivo studies should be performed to explore CNDs and their behavior in different biological systems.

Day by day, several innovative applications of mushrooms can be noticed in their farming and cultivation. These applications mainly focus on the medicinal and prebiotic attributes, besides their role for producing the food and energy. This mini-review is an attempt to highlight the cultivation of mushrooms and their requirements under different kinds of farming of mushrooms. These farming types may include mushroom-worm system, mushroom-bee farming, smart mushroom farming, forest-mushrooms farming, urban mushroom farming, and mushroom-livestock farming. This study included the condition of cultivation of several mushroom species and their edibility. Edible/medicinal mushrooms are well-known with their high content of many beneficial bioactive ingredients for human health such as antioxidants, ergosterols, lectins, phenolics/polyphenolics, polysaccharides, and terpenoids. These bioactives of mushrooms have a potential capability to treat and/or prevent several chronic diseases. The medicinal and prebiotic attributes of mushrooms still need more investigations, and this topic of "mushrooms in medicine" stills have several open questions are needed to be answer in the future.

Walnut (Juglans regia L.) production is a developing sector in Hungarian horticulture, where preharvest fruit rot increasingly causes economic losses. Previously, fungi from the Diaporthaceae and Botryosphaeriaceae families were isolated with a high incidence from rotten fruits. Here, we identify these pathogens from different plant parts (twigs, buds, and shoots) in orchards in the north-east of Hungary, and test their pathogenicity on immature nuts. Diaporthe eres Nitschke and Diplodia seriata De Notaris were identified in isolates from different symptomatic and asymptomatic plant parts based on their ITS (nuclear ribosomal internal transcribed spacer) and tef1 (translation elongation factor EF-1-alpha gene) DNA sequences. Purified monocultures caused rot of immature nuts following in vitro inoculation. Our results suggest that D. eres Nitschke and D. seriata De Notaris lingering on buds and overwintering woody parts may affect the seasonal nuts through wound infection. Infection by Diaporthe and Botryosphaeriaceae species present on woody plant parts, as well as on/in buds has been reported in Mediterranean countries. This is the first report of such stepwise aetiology from a region with continental weather. Climate change, associated weather patterns and the limitations of fungicide use in the European Union, among other factors, could be responsible for the increasing number of infections and economic damage caused by these pathogens.

MAILLARD reactions (MRs) is a non-enzymatic browning process, which helps to produce flavor and aroma, and different compounds with different health effects in the food and feed. The reaction is between reducing carbohydrates (reducing sugars) and amino compounds and it can be classified into three stages (initial, intermediate, and final). Depending on the reaction and storage conditions, activity and complexity of reactants used tools for environment control, and applied biotechnological applications, both beneficial and toxic MRPs can be produced. This review aims to analyze the activities and mechanisms of components forming by the Maillard reaction and the most common bioactive Maillard reaction products (MRPs) with prebiotic, antifungal, and antibacterial activity. Moreover, the study of the possible formation of toxic chemicals and having a look at the elimination ways of this potential problem was also an important goal in this review. The main aim of this review is to address the benefits of Maillard reaction products, which have been identified in previous studies to have antimicrobial and prebiotic effects, for the treatment of human pathogens. The present study will help to understand the positive and negative sides of Maillard reaction.

In the first part of this two-piece publication, the isolation, identification and in vitro characterization of ten endophytic Trichoderma isolates were reported. Here we report the ability of two different mixes of some of these isolates (Trichoderma simmonsii, Trichoderma orientale and Trichoderma gamsii as well as of Trichoderma afroharzianum and T. simmonsii) to colonize and stimulate the growth of grapevines. Two commercial vineyards about 400 km away from the site of isolation were used as experimental fields, from which the strains of three Trichoderma species were re-isolated up to four years after rootstock soaking treatment with conidiospores, performed before planting. The treatments decreased the overall percentage of lost plants of about 30%, although a low number of lost plants (about 5%) were observed also in the control plot. For all cultivars and clones, the Trichoderma treatments significantly increased both the bud burst ratio and bud burst vigor index. In addition, the grape must parameters such as the Brix degrees, as well as the extract, the D-glucose and the D-fructose concentrations all appeared to be improved, suggesting a potentially higher ethanol content of the produced wine. We conclude that grapevine-endophytic Trichoderma isolates promote plant growth, which could be a useful feature for sustainable agriculture in general and integrated plant production in particular.

Pleurotus ostreatus (Jacq. ex Fr.) P. Kumm mushroom contains bioactive compounds with both antimicrobial and prebiotic properties, which are distributed in the mushroom mycelium, fruiting body, and spent substrate. The mushroom is rich in nondigestible carbohydrates like chitin and glucan, which act as prebiotics and support the growth and activity of beneficial gut bacteria, thereby maintaining a healthy balance of gut microbiota and reducing the risk of antibiotic resistance. The bioactive compounds in P. ostreatus mushrooms, including polysaccharides (glucans, chitin) and secondary metabolites (phenolic compounds, terpenoids, and lectins), exhibit antibacterial, antiviral, and antifungal activities. When mushrooms are consumed, these compounds can help preventing the growth and spread of harmful bacteria in the gut, reducing the risk of infections and the development of antibiotic resistance. Nonetheless, further research is necessary to determine the efficacy of P. ostreatus against different pathogens and to fully comprehend its prebiotic and antimicrobial properties. Overall, consuming a diet rich in mushroom-based foods can have a positive impact on human digestion health. A mushroom-based diet can support a healthy gut microbiome and reduce the need for antibiotics.

Aflatoxin contamination can appear in various points of the food chain. If animals are fed with contaminated feed, AFB1 is transformed—among others—to aflatoxin M1 (AFM1) metabolite. AFM1 is less toxic than AFB1, but it is still genotoxic and carcinogenic and it is present in raw and processed milk and all kinds of milk products. In this article, the chronic exposure estimation and risk characterization of Hungarian consumers are presented, based on the AFM1 contamination of milk and dairy products, and calculated with a probabilistic method, the two-dimensional Monte-Carlo model. The calculations were performed using the R plugin (mc2d package) integrated into the KNIME (Konstanz Information Miner) software. The simulations were performed using data from the 2018–2020 food consumption survey. The AFM1 analytical data were derived from the Hungarian monitoring survey and 1,985 milk samples were analyzed within the framework of the joint project of the University of Debrecen and the National Food Chain Safety Office of Hungary (NÉBIH). Limited AFM1 concentrations were available for processed dairy products; therefore, a database of AFM1 processing factors for sour milk products and various cheeses was produced based on the latest literature data, and consumer exposure was calculated with the milk equivalent of the consumed quantities of these products. For risk characterization, the calculation of hazard index (HI), Margin of Exposure, and the hepatocellular carcinoma incidence were used. The results indicate that the group of toddlers that consume a large amount of milk and milk products are exposed to a certain level of health risk. The mean estimated daily intake of toddlers is in the range of 0.008–0.221 ng kg−1 bw day−1; the 97.5th percentile exposure of toddlers is between 0.013 ng kg−1 bw day−1 and 0.379 ng kg−1 bw day−1, resulting in a HI above 1. According to our study, the exposure of older age groups does not pose an emergent health risk. Nevertheless, the presence of carcinogenic compounds should be kept to a minimum in the whole population.

Aflatoxins (AFs) are toxic secondary metabolites produced mostly by Aspergillus species. AF contamination entering the feed and food chain has been a crucial long-term issue for veterinarians, medicals, agroindustry experts, and researchers working in this field. Although different (physical, chemical, and biological) technologies have been developed, tested, and employed to mitigate the detrimental effects of mycotoxins, including AFs, universal methods are still not available to reduce AF levels in feed and food in the last decades. Possible biological control by bacteria, yeasts, and fungi, their excretes, the role of the ruminal degradation, pre-harvest biocontrol by competitive exclusion or biofungicides, and post-harvest technologies and practices based on biological agents currently used to alleviate the toxic effects of AFs are collected in this review. Pre-harvest biocontrol technologies can give us the greatest opportunity to reduce AF production on the spot. Together with post-harvest applications of bacteria or fungal cultures, these technologies can help us strictly reduce AF contamination without synthetic chemicals.

A methicillin-rezisztens Staphylococcus aureus (MRSA) törzsek élelmiszerláncban előforduló jelenlétét számos tanulmány igazolta az Európai Unióban, azonban Magyarországon kevés adat áll rendelkezésünkre ezzel kapcsolatban. Jelen vizsgálat célja az élelmiszerekből izolált Staphylococcus törzsek antibiotikum rezisztenciá-jának vizsgálata klasszikus mikrobiológiai, molekuláris biológiai módszerekkel és MALDI-TOF-MS technikával, továbbá az antibiotikum rezisztens törzsek multilókusz szekvencia tipizálása (MLST). A vizsgálat során 47 koaguláz-pozitív (CPS) és 30 koaguláz-negatív Staphylococcus (CNS) izolátumot gyűjtöttünk. A MALDI-TOF-MS vizsgálat során minden CPS izolátum (n=47) S. aureus fajnak bizonyult, míg a CNS törzsek esetében 8 különböző fajt azonosítottunk. Két S. aureus törzs esetében állapítottunk meg methicillin-rezisztenciát, amelyek közül az egyik izolátum eddig még nem ismert szekvencia típusba, míg a másik MRSA törzs az ST398 típusba tartozott, amely a mezőgazdasági haszonállatokból izolált MRSA törzsek leggyakoribb típusa az EU/EGT területén. (Az „MRSA” rövidítést köznapi szóhasználatban, de esetenként a szakirodalomban is gyakran a „multirezisztens Staphylococcus aureus” megjelölésére használják. A szerzők kéziratában - helyesen a methicillin-rezisztens kórokozót jelölik így. A Szerk.)

The objective of this study was to use matrix-assisted laser desorption ionisation–time of flight mass spectrometry (MALDI-TOF MS) for the identification of ovine-associated staphylococci. Presumptive Staphylococcus isolates were recovered from ovine udder surface (US), individual raw milk, bulk tank milk, and cheese samples and were characterised by conventional phenotypic methods. A total of 69 bacterial isolates were further confirmed by MALDI-TOF MS. Forty-two (60.9%) of 69 isolates were successfully identified on genus and species level. Two thirds (n = 28) of the 42 identified isolates were shown to be Staphylococcus spp. These 28 staphylococcal isolates formed two clusters, one consisting of 22 Staphylococcus aureus strains and the other composed of 6 non-aureus staphylococci, including S. simulans (n = 3), S. auricularis, S. equorum, and S. haemolyticus. MALDI-TOF MS has proven to be a reliable tool for the identification of staphylococci from raw sheep's milk, especially bulk tank milk; however, currently it appears to be less useful for the identification of bacterial isolates originating from ovine US samples. This is the first study to evaluate the applicability of MALDI-TOF MS for identification of Staphylococcus spp. in ovine raw milk, cheese, and US samples in Hungary.

Aflatoxins (AFs) are among the most harmful fungal secondary metabolites imposing serious health risks on both household animals and humans. The more frequent occurrence of aflatoxins in the feed and food chain is clearly foreseeable as a consequence of the extreme weather conditions recorded most recently worldwide. Furthermore, production parameters, such as unadjusted variety use and improper cultural practices, can also increase the incidence of contamination. In current aflatoxin control measures, emphasis is put on prevention including a plethora of pre-harvest methods, introduced to control Aspergillus infestations and to avoid the deleterious effects of aflatoxins on public health. Nevertheless, the continuous evaluation and improvement of post-harvest methods to combat these hazardous secondary metabolites are also required. Already in-use and emerging physical methods, such as pulsed electric fields and other nonthermal treatments as well as interventions with chemical agents such as acids, enzymes, gases, and absorbents in animal husbandry have been demonstrated as effective in reducing mycotoxins in feed and food. Although most of them have no disadvantageous effect either on nutritional properties or food safety, further research is needed to ensure the expected efficacy. Nevertheless, we can envisage the rapid spread of these easy-to-use, cost-effective, and safe post-harvest tools during storage and food processing.

A tej összetételének és higiéniai tulajdonságainak változásai befolyásolják a termelői árat, ezért elengedhetetlen, hogy a felelős tejtermelő információt gyűjtsön ezen paraméterek különböző tényezők hatására bekövetkező változásairól. A szerzők a tanulmányukban arra keresik a választ, hogy tapasztalható-e változás a tehenek napi tejmennyiségében, illetve a nyers tehéntej összetételében (zsír- és fehérjetartalom) és mikrobiológiai tulajdonságaiban (szomatikus sejtszám, összcsíraszám, coliform- és S. aureus-szám) az egyszer és többször ellett teheneknél, illetve a laktációjuk különböző stádiumaiban. Egy hazai nagyüzemi tejtermelő telep adatai alapján megállapították, hogy a tej zsír- és fehérjetartalmában ugyan nem volt különbség, de a többször ellett teheneknél nagyobb volt a napi tejhozam, valamint az egyszer ellett tehenek tejéhez képest a tejben magasabb volt a szomatikus sejtszám és nagyobb mennyiségben fordultak elő coliform baktériumok. A napi tejmennyiség a laktáció egymást követő stádiumaiban csökkent, viszont a tej zsír- és fehérjetartalma növekedést mutatott, amely feltételezhetően a csökkenő tejmennyiség koncentráló hatásának tulajdonítható. A mikroorganizmusok telepszámában a laktáció különböző stádiumaiban nem tapasztaltak jelentős változást.

Changes in the composition and hygienic properties of milk affect producer price, so it is essential for the responsible dairy farmer to collect information on changes in these parameters due to various factors. In their study, the authors seek to answer the question whether there is a fluctuation in the daily milk yield of cows and in the composition (fat and protein content) and microbiological properties (somatic cell count, total plate count, coliform and S. aureus count) of raw cow’s milk in primiparous and multiparous cows or at different stages of their lactation. Based on the data of a Hungarian large-scale dairy farm, it was found that there was no difference in the fat and protein content of the milk, but the daily milk yield was higher in the case of multiparous cows and, compared to the milk of primiparous cows, their milk had a higher somatic cell count and larger amounts of coliform bacteria. The daily milk yield decreased in the successive stages of lactation, but the fat and protein content of the milk increased, which is presumably due to the concentrating effect of the decreasing milk yield. No significant change was observed in the colony count of microorganisms at the different stages of lactation.

In this research, our aim was to assess the occurrence of Staphylococcus aureus in a Hungarian large-scale dairy farm during the S. aureus control program conducted in the course of our studies. Furthermore, the phenotypic and genotypic properties of the isolates (type of haemolysis, antibiotic susceptibility, staphylococcal enterotoxin (SE) gene carrying ability and spa type) were determined. S. aureus was detected in all bulk tank milk samples collected during this study. Two different spa types were identified among the 17 strains isolated in the farm. A total of 14 of the 17 studied strains (82%) showed β-haemolysis on blood agar, 2/17 strains (12%) expressed double zone and 1/17 strains (6%) showed weak β-haemolysis. All strains were susceptible to most antibiotics tested (cefoxitin, chloramphenicol, clindamycin, erythromycin, gentamicin, tetracycline and trimethoprim/sulphamethoxazole), but all strains were resistant to penicillin G. A total of 11 of the 17 strains (65%) were found to harbour seg, sei, selm, seln, selo genes; 4/17 strains (24%) harboured sei, selm, seln, selo genes and 2/17 strains (11%) harboured sei gene. Since the new SEs/SEls can also cause foodborne outbreaks potentially and all strains were found to be resistant to penicillin G, it is essential to decrease and keep the prevalence of S. aureus low in the dairy farm and the implementation of the S. aureus control program is also highly justified. The results showed that the S. aureus count decreased by the end of our studies, so the control program was proved to be effective.

Fungal mycotoxins are secondary metabolites that can be present in green forage, hay, or silage. Consumption of contaminated plants or agricultural products can cause various animal and human diseases, which is why problems associated with mycotoxins have received particular attention. In addition, public pressure to produce healthy food and feed is also increasing. As the results of several surveys indicate that yeasts can decrease toxic effects by binding or converting secondary metabolites or control growth of harmful fungi, this article provides an overview of the yeast species that can have great potential in detoxification. The most important antagonistic yeast species against toxigenic fungi are described and the mode of their inhibitory mechanisms is also discussed. We provide an insight into toxin binding and biotransformation capacities of yeasts and examples of their use in silo. Issues requiring further study are also mentioned.