The inhibitory effects of plant additives on biogenic amine formation in fermented foods - a review.

Fermented food has unique properties and high nutritional value, and thus, should constitute a basic element of a balanced and health-promoting diet. However, it can accumulate considerable amount of biogenic amines (BAs), which ingested in excess can lead to adverse health effects. The application of plant-derived additives represents a promising strategy to ensure safety or enhance the functional and organoleptic properties of fermented food. This review summarizes currently available data on the application of plant-origin additives with the aim to reduce BA content in fermented products. The importance of ensuring fermented food safety has been highlighted considering the growing evidence of beneficial effects resulting from the consumption of this type of food, as well as the increasing number of individuals sensitive to BAs. The examined plant-origin additives reduced the BA concentration to varying degrees, and their efficacy depended on the type of additive, matrix, autochthonous, and inoculated microorganisms, as well as the manufacturing conditions. The main mechanisms of action include antimicrobial effects and the inhibition of microbial decarboxylases. Further research on the optimization of bioactive substances extraction, standardization of their chemical composition, and development of detailed procedures for its use in fermented products manufacturing are needed.

Thyme essential oil inhibits intracellular replication of CyHV-3 and inactivates extracellular virus. An in vitro study.

Cyprinid herpesvirus 3 (CyHV-3) can induce up to 100% mortality among carp populations. To date, there has been no safe method to prevent the consequences of the activity of CyHV-3. Thyme is widely used in cooking due to its flavour. Both thyme and thyme essential oil (TEO) are used in traditional herbal medicine, mainly to treat respiratory system disorders. In this study, TEO containing predominantly cymene and thymol was applied to explore its antiviral effect. The toxicity of TEO was examined in MTT and crystal violet assays. The anti-CyHV-3 activity of TEO in the intracellular and extracellular stages of the viral replication cycle was explored in a plaque assay and TaqMan qPCR. TEO interfered with the intracellular stages of the CyHV-3 replication cycle with selectivity indexes (SI) of around 5. It also displayed virucidal activity in a dose- and time-dependent manner. Two-hour preincubation of CyHV-3 with TEO generated SI, ranging from 13.37 to 18.47 depending on cell line and method of examination. Preincubation of cells with TEO at a safe concentration did not decrease the intracellular viral DNA copy number, which suggests that TEO does not disturb the attachment of the virus to the cells. Further research regarding the antiviral activity of compounds of TEO is required in order to indicate the most potent molecules that could be considered candidates for application in aquaculture.

The Development, Validation, and Application of a UHPLC-HESI-MS Method for the Determination of 17 Cannabinoids in Cannabis sativa L. var. sativa Plant Material.

Cannabinoids are an important group of secondary metabolites found in the plant Cannabis sativa L. The growing interest in the use of hemp in food production (e.g., hemp teas, hemp cookies) makes it necessary to develop a method for determining these compounds in the plant, both fresh and dried. The selection of a suitable extraction liquid for the extraction of cannabinoids and the development of a method for the determination of 17 cannabinoids is a prelude to the development of an effective method for the extraction of these compounds. In the present study, a novel, simple, and efficient method was developed and validated for the determination of up to 17 cannabinoids in fresh plant parts (inflorescences and leaves) of Cannabis sativa L. and in dried material, including hemp teas. Analyses were performed using ultra-high-performance liquid chromatography-Q-Exactive Orbitrap mass spectrometry setup operating with a heated electrospray interface (UHPLC-HESI-MS). Based on the comparison, methanol was selected as the best for the extraction of cannabinoids from fresh and dried material. The efficiency and validity of the method were assessed using certified reference material (dried Cannabis) and confirmed by z-score from participation in an international proficiency test conducted by ASTM International for dried hemp.

Changes in the microbiological quality and content of biogenic amines in chicken fillets packed using various techniques and stored under different conditions.

The aim of this study was to determine the biogenic amines (BAs) formed in chicken breast meat packaged using different techniques (AP, Hi-O2-MAP or VP) during the storage under different conditions (cold room or display case), to correlate the microbiological quality (TPC, LAB, Pseudomonas spp. and Enterobacteriaceae) of chicken meat with BAs formation and to assess the suitability of selected biogenic amines as indicators of chicken meat spoilage. The initial TPC of chicken fillets was 2.57-3.04 log cfu/g. Over time a systematic significant (p ≤ 0.05) increase in TPC was observed to >7.5 log cfu/g (AP and VP; display case) determined on day 9. It was found that cadaverine and tyramine dominated in quantitative terms in chicken fillets, regardless of packaging technique and storage conditions (166.00 mg/kg in AP meat in cold room on day 9 and 175.03 mg/kg on day 9 in MAP meat in display case, respectively). Taking into account the BAI, high and significant (p ≤ 0.05) correlation coefficients (from 0.51 to 0.95) were obtained with all analyzed indicators of microbiological quality. The concentration of cadaverine, putrescine contents or BAI can potentially serve as chemical quality indicator for freshness of chicken meat.

Cannabinoids-Characteristics and Potential for Use in Food Production.

Scientific demonstrations of the beneficial effects of non-psychoactive cannabinoids on the human body have increased the interest in foods containing hemp components. This review systematizes the latest discoveries relating to the characteristics of cannabinoids from Cannabis sativa L. var. sativa, it also presents a characterization of the mentioned plant. In this review, we present data on the opportunities and limitations of cannabinoids in food production. This article systematizes the data on the legal aspects, mainly the limits of Δ9-THC in food, the most popular analytical techniques (LC-MS and GC-MS) applied to assay cannabinoids in finished products, and the available data on the stability of cannabinoids during heating, storage, and access to light and oxygen. This may constitute a major challenge to their common use in food processing, as well as the potential formation of undesirable degradation products. Hemp-containing foods have great potential to become commercially popular among functional foods, provided that our understanding of cannabinoid stability in different food matrices and cannabinoid interactions with particular food ingredients are expanded. There remains a need for more data on the effects of technological processes and storage on cannabinoid degradation.

Time Evolution of Microbial Composition and Metabolic Profile for Biogenic Amines and Free Amino Acids in a Model Cucumber Fermentation System Brined with 0.5% to 5.0% Sodium Chloride.

Salt concentrations in brine and temperature are the major environmental factors that affect activity of microorganisms and, thus may affect formation of biogenic amines (BAs) during the fermentation process. A model system to ferment cucumbers with low salt (0.5%, 1.5% or 5.0% NaCl) at two temperatures (11 or 23 °C) was used to study the ability of indigenous microbiota to produce biogenic amines and metabolize amino acid precursors. Colony counts for presumptive Enterococcus and Enterobacteriaceae increased by 4 and up to 2 log of CFU∙mL-1, respectively, and remained viable for more than 10 days. 16S rRNA sequencing showed that Lactobacillus and Enterobacter were dominant in fermented cucumbers with 0.5% and 1.5% salt concentrations after storage. The initial content of BAs in raw material of 25.44 ± 4.03 mg∙kg-1 fluctuated throughout experiment, but after 6 months there were no significant differences between tested variants. The most abundant BA was putrescine, that reached a maximum concentration of 158.02 ± 25.11 mg∙kg-1. The Biogenic Amines Index (BAI) calculated for all samples was significantly below that needed to induce undesirable effects upon consumption. The highest value was calculated for the 23 °C/5.0% NaCl brine variant after 192 h of fermentation (223.93 ± 54.40). Results presented in this work indicate that possibilities to control spontaneous fermentation by changing salt concentration and temperature to inhibit the formation of BAs are very limited.

Background levels of polycyclic aromatic hydrocarbons and legacy organochlorine pesticides in wheat sampled in 2017 and 2018 in Poland.

Both polycyclic aromatic hydrocarbons (PAHs) and legacy organochlorine insecticides (OCPs), including DDT, are dangerous chemical contaminants. The aims of this study were to (i) determine background levels of PAHs and legacy OCPs for wheat samples collected in 2017 and 2018 in Poland, (ii) identify differences between levels in wheat harvested in various regions of Poland, (iii) evaluate differences in contamination sources manifested by the profiles of the identified chemicals, (iv) identify possible correlations between different classes of chemicals present in wheat, and (v) assess the health risks associated with the presence of PAHs and OCPs in Polish wheat. Average concentrations found in the samples were 0.09 ± 0.03 µg kg-1 for benzo[a]pyrene (BaP) (formerly used as a single PAH marker), 0.43 ± 0.16 for the more recently introduced collective PAH 4 marker (benzo[a]anthracene + benzo[a]pyrene + chrysene + benzo[b]fluoranthene), and 1.07 ± 0.68 µg kg-1 for DDT and its metabolites. The PAH profiles indicated contamination from combustion-related emission sources (liquid fossil fuels, coal, biomass). Health risks associated with the presence of PAHs and OCPs in cereals were assessed using the margin of exposure (MOE) approach. The MOE values calculated based on the highest concentrations found in this study exceeded 50,000 for both BaP and PAH 4. The calculated worst-case scenario value for DDT and metabolites was as low as 0.3% of the respective tolerable daily intake (TDI) value. Assessment of dietary risk has shown that the presence of the two contaminant classes in Polish wheat grains is of low concern.

Dietary risk evaluation for 28 polycyclic aromatic hydrocarbons (PAHs) in tea preparations made of teas available on the Polish retail market.

The aim of this work was to assess dietary risk resulting from consumption of polycyclic aromatic hydrocarbons (PAHs) with tea infusions. To this end, levels of 28 PAHs in black, green, red and white teas available on the Polish retail market have been assessed. Profiles and correlation between concentrations of individual PAHs have been identified. A model study on transfer of PAHs from tea leaves into tea preparations has been conducted. Relatively high concentrations of 28 evaluated PAHs have been found in 58 tested samples of black, green, red and white teas sampled on the Polish retail market. Total concentration ∑28PAH ranged from 57 to 696 µg kg-1 with mean 258 µg kg-1 (dry tea leaves). The most mature tea leaves fermented to a small degree contained relatively the highest PAH levels among all four tested tea types. Relatively low PAH transfer rates into tea infusions and limited volumes of the consumed tea keep the risks associated with PAH dietary intake at a safely low level. The worst-case scenario dietary intake values were 7.62/0.82/0.097 ng kg-1 b.w. day-1 (estimated on the basis of the maximum found concentrations 696/113/23 µg kg-1 and maximum observed transfer rates 24/16/9%) for ∑28PAH/∑PAH4/B[a]P, respectively. MOE values calculated using the above worst case estimates exceeded 700,000 and 400,000 (BMDL10 0.07 and 0.34 mg kg-1 b.w. day-1) for B[a]P and PAH4, respectively. Both B[a]P and PAH4 concentrations may be used as indicators of total PAH concentration in tea leaves; PAH4 slightly better fits low molecular weight PAHs. Several correlations between various PAHs/groups of PAHs have been identified, the strongest one (R2 = 0.92) between PAH4 and EU PAH 15+1.

Optimized yeast-based in vitro bioassay for determination of estrogenic and androgenic activity of hydroxylated / methoxylated metabolites of BDEs / CBs and related lipophilic organic pollutants.

Persistent organic pollutants (POPs) are known to show endocrine disrupting (ED) activity, including interactions with hormone receptors. The aim of this work was to develop a bioassay applicable for evaluation of ED potency of highly lipophilic metabolites of POPs. To that end, a yeast-based bio-assay protocol was used. Estrogenic / androgenic activity of some native brominated biphenyl ethers (BDEs) / chlorinated biphenyls (CBs), and their hydroxylated / methoxylated metabolites was assessed. Since data (including potency compared to reference native hormones) obtained using different protocols vary, the possibility that yeast transforms POPs into some more potent compounds was first checked; it seems that no such transformation is important from the test applicability standpoint. The developed method was sensitive with EC50 values 6.5*10-11 M and 4.5*10-9 M calculated for E2 and DHT, respectively. Both CBs and BDEs show weak estrogenic activity negatively correlated with the degree of their halogenation, but their metabolites are significantly more potent xenohormones. 4-OH-2,2',4',6'-TeCB was the most potent estrogen receptor (ER) agonist among all tested compounds; its activity was only 1,000 times lower than that of native E2.

Application of Liquid Chromatography/Ion Trap Mass Spectrometry Technique to Determine Ergot Alkaloids in Grain Products.

A liquid chromatography/ion trap mass spectrometry-based method to determine six ergot alkaloids and their isomers is presented. The samples were cleaned on neutral alumina-based solid-phase extraction cartridges. The following method parameters were obtained (depending on the analyte and spiking level): method recovery from 63.0 to 104.6%, relative standard deviation below 18%, linear range from 1 to 325 µg/kg, linear correlation coefficient not less than 0.98. The developed analytical procedure was applied to determine the levels of ergot alkaloids in 65 samples of selected rye-based food products (flour - 34 samples, bran - 12 samples, rye - 18 samples, flakes - 1 sample). Measurable levels of alkaloids were found in majority of the analysed samples, particularly in rye flour. Additionally, alkaloids were determined in ergot sclerotia isolated from rye grains. Total content was nearly 0.01% (97.9 mg/kg). However, the alkaloid profile was dominated by ergocristine at 45.6% (44.7 mg/kg), an alkaloid not commonly found in the tested food products. Ergocorninine at 0.2% (0.2 mg/kg) was the least abundant alkaloid.

Fate of PBDEs during food processing: Assessment of formation of mixed chlorinated/brominated diphenyl ethers and brominated dioxins/furans.

The aim of this study was to evaluate effects of food processing on PBDE levels, in particular influence of heat treatment on degradation of PBDEs, including possible formation of chlorinated diphenyl ethers or brominated dioxins/furans as degradation products. It was shown that PBDEs heated in the presence of chlorine (from either organic or inorganic sources) formed mixed chlorinated/brominated diphenyl ethers. However, no PCDEs were formed in the presence of lipids. Lipid medium increased stability of PBDEs exposed to UV irradiation. Profile of congeners formed in result of the debromination reaction was significantly different than profiles observed by some other authors in aliphatic organic solvents. Grilling processes increased concentrations (calculated on the fresh product basis) of the studied compounds by 4-8/22-34% for electric/coal grill, respectively. Depending on the congener and on the applied heat treatment, PBDE mass in pork meat after grilling dropped by 26-53%. No detectable quantities of either brominated dioxins or furans were formed during thermal processing of food containing typical levels of PBDEs.

Intersex gonad differentiation in cultured Russian (Acipenser gueldenstaedtii) and Siberian (Acipenser baerii) sturgeon.

Among sturgeons, the occurrence of individuals with gonads containing both testis and ovary components is considered pathological, and such fish are described as intersex individuals or intersexes. Intersexes are observed in both wild and cultured populations of sturgeon, usually at low frequencies. In the present study, intersex Russian (Acipenser gueldenstaedtii) and Siberian (Acipenser baerii) sturgeons constituted 30% of the studied populations. Macroscopically, intersex gonads were recognizable from 500 days posthatching (dph). Initially, gonads with predominantly male characteristics (testis-ova) were observed, but in older fish gonads with predominantly female traits (ova-testis) were more frequent. Using microscopic analysis, intersex gonads were discernible by 130-200 dph. Observations of intersex germinal epithelium development and analysis of sex distribution in the study populations indicated that feminization was occurring. Histological analysis revealed that differentiation of the germinal epithelium in such gonads was accompanied by various morphological alterations (transformations) that were described using quantitative and localization criteria. The most common type of transformations, massive subepithelial transformations, was manifested by the presence of abundant female germinal tissue located under the gonad surface epithelium in the developing testis. These transformations were identified in the early development stage (100-200 dph). In this type of transformation, differentiation of female germinal tissue at the gonad surface and male tissue at the mesorchium/mesovarium resulted in complete formation of both male and female germinal epithelia within the same gonad.

Seasonal variability of polychlorinated biphenyls (PCBs) and polychlorinated diphenyl ethers (PBDEs) congener profiles in butter in Poland: dietary risk evaluation.

Various statistical methods have been employed to analyse in details seasonal diversification of polychlorinated biphenyl (PCB)/polybrominated diphenyl ether (PBDE) congener profiles found in butter fat. The variability of the PCB/PBDE congener profiles indicates the presence of various sources of the milk fat contamination. The obtained results suggest that the environmental chemical background has the highest share in the contamination sources pattern. Ion trap mass spectrometry coupled to high-resolution gas chromatography with semi-permeable membrane dialysis sample cleanup was used for determination of PCBs and PBDEs in milk fat. Determined butter fat PCB profiles were similar to the profiles characteristic for Aroclor 1254 technical mixture. Our data indicate that dietary intake of PCB/PBDE with milk and milk products may be estimated to be about 717.5 pg kg b.w.(-1) day(-1) for six-indicator PCBs, 0.329 (equivalent toxicity, TEQ) pg kg b.w.(-1) day(-1) for 12 DL PCBs and 50 pg kg b.w.(-1) day(-1) for PBDEs (sum of 14 congeners).

Heterogeneity of transport and metabolism of Tormentillae rhizoma constituents across human intestinal epithelium cellular model.

Tormentilla erecta (L.) Raeusch is a widespread plant in Europe and Western Asia. Its rhizomes (Tormentilae rhizoma) are the main ingredient of herbal alcoholic beverages and can be used as a natural preservative in beer production. Apart from its unique taste qualities, therapeutic properties in gastrointestinal tract ailments are attributed to the tincture obtained from Tormentillae rhizoma. The presented research aimed to determine the mutual relationship between the components of Tormentillae tincture, present in popular alcoholic beverages, and intestinal epithelium (Caco-2 cell monolayers). A comprehensive qualitative and quantitative analysis of the tincture was performed, including the determination of condensed and hydrolyzable tannins as well as triterpenoids (UHPLC-DAD-MS/MS). Incubation of the tincture with Caco-2 monolayers has shown that only triterpenes pass through the monolayer, while condensed tannins are mainly bound to the monolayer surface. Ellagic acid derivatives were the only components of the Tormentillae tinctura being metabolized by cell monolayers to the compounds not previously described in the literature, which may be crucial in the treatment of intestinal diseases with inflammatory background.

Evaluating birch leaf tea as a functional herbal beverage: Beneficial impact on the urinary tract, and metabolism in human organism.

Herbal teas are considered as a potential constituent of novel functional beverages consumed daily. One of the commonly used herbal teas is silver birch (Betula pendula Roth) leaf infusion, traditionally used in urinary tract diseases. In this study, the potential of birch leaf infusion as a functional beverage, emphasizing its active ingredients' bioavailability, anti-inflammatory, and antiadhesive properties concerning urinary tract health, was investigated. A complex approach was proposed, which included phytochemical screening, bioavailability, gut microbiota biotransformation, and an in vivo test for urine metabolomics assessment. The bioassays confirmed significant anti-inflammatory (interleukins IL-6 and IL-8 secretion) and anti-adhesive (Uropathogenic Escherichia coli and T24 bladder cells) activities. The high-resolution mass spectrometry metabolomics studies linked gut microbiota metabolites and the metabolites present in the urine. Several metabolites connected with phenolics' consumption were detected in the urine, e.g., glucuronides and sulfates of caffeic acid and dihydroxyphenyl-γ-valerolactones. Based on the presented results, the birch leaf should be considered useful in designing functional beverages, especially targeted to the groups at high risk of urinary diseases.

Content of amino acids and biogenic amines in stored meat as a result of a broiler diet supplemented with ß-alanine and garlic extract.

Poultry meat is a highly esteemed product among consumers. However, the emphasis on increasing body weight has led to a rise in the proportion of rapidly shrinking fibers, adversely affecting the quality and shelf life of poultry meat. With a growing awareness of dietetics among consumers, there is an increasing challenge to produce chicken meat that is not only free of antibiotics but also beneficial for dietary and health reasons. Biogenic amines (BA) can serve as indicators of meat freshness and quality. While they play vital roles in the body, excessive consumption of BA can have toxic and carcinogenic effects. The objective of this study was to examine the impact of supplementing feed with garlic extract and ß-alanine (ß-Ala) on the formation of BA and amino acid (AA) levels in the breast and leg muscles of chickens stored under aerobic chilling conditions. The muscles were obtained from chickens fed with garlic extract and ß-Ala in quantities of 0.5 and 2% for each additive, as well as 0.5 and 2% of their combination. Analyses were conducted on d 1, 3, 5, 7, and 10 of storage. ß-Ala supplementation increased the proportion of this AA in breast (P < 0.01) and leg muscles (P < 0.01), along with a rise in the proportion of nonessential AA (NEAA; sum of aspartic, aspartic acid, glutamic, glutamic acid, serine, ß-Ala, and proline) (P < 0.01). The levels of BA changed during storage in breast and leg muscles (P < 0.001). The applied diet significantly influenced the formation of putrescine (P = 0.030), phenylethylamine (P = 0.003), agmatine (P = 0.025), and total BA (P < 0.001) in breast muscles. On the 10 d of storage, the breast muscles exhibited the lowest BA index (BAI) in the group, with a diet supplemented with 0.5% garlic extract and 0.5% ß-Ala (P < 0.05). The leg muscles showed a similar BA trend as the breast muscles. These supplements may be utilized in production to augment the protein content of chicken muscles and potentially decrease the BAI index during meat storage.

Application of molecularly imprinted polymers to determine B1, B2, and B3 fumonisins in cereal products.

The aim of this study was to develop an analytical method for qualitative and quantitative determination of the B(1), B(2,) and B(3) fumonisins in cereal products. A LC coupled to an IT-MS was used as the analytical instrument. The AFFINIMIP FumoZON Molecularly Imprinted Polymer SPE cartridges (Polyintell) were used to isolate fumonisins from the analyzed samples and the clean-up step. Statistical parameters evaluated in some validation experiments were as follows: mean recovery 95-106%, precision <17% (expressed as recovery RSD). The developed method was used to determine fumonisins in 49 cereals (42 maize-based and seven wheat-based products). In most cases, concentrations of the studied compounds found in the analyzed samples were low. The highest total concentration of the B(1), B(2), and B(3) fumonisins was found in maize flour samples (range, 26-1102 µg/kg, mean 498 µg/kg).

Chemical Stability of the Lipid Phase in Concentrated Beverage Emulsions Colored with Natural ß-Carotene.

The aim of this study was to examine the oxidation of selected plant oils in concentrated beverage emulsions colored with natural ß-carotene. Carotenoid preparations obtained from carrots were dissolved in cold-pressed linseed oil, refined canola oil, and refined palm olein. Oxidative stability of the lipids was examined with and without addition of the pigment to the oil/water (O/W) emulsion. Carotenoid/lipid hydro peroxide (LOOH) concentration was evaluated using two different methods: LOOH + Fe2+ reaction connected with a colored complex of ammonium thiocyanate determined with the help of a spectrophotometer, and LOOH determined with the help of a chemiluminometer. It was shown that oxidation rate of lipids in the O/W emulsions strongly depended on chemical composition of the lipid fraction (type of oil used). Presence of the carotenoid pigment increased the rate. Therefore, if a carotenoid-containing emulsion is to be stable, it should be based on oils of a high oxidative stability.

Recent advances and opportunities related to the use of bee products in food processing.

Nowadays, natural foods that can provide positive health effects are gaining more and more popularity. Bees and the products they produce are our common natural heritage that should be developed. In the article, we presented the characteristics of bee products and their use in industry. We described the development and importance of beekeeping in the modern world. Due to their high nutritional value and therapeutic properties, bee products are of great interest and their consumption is constantly growing. The basis for the use of bee products in human nutrition is their properties and unique chemical composition. The conducted research and opinions confirm the beneficial effect of bee products on health. The current consumer awareness of the positive impact of food having a pro-health effect on health and well-being affects the increase in interest and demand for this type of food among various social groups. Enriching the daily diet with bee products may support the functioning of the organism. New technologies have appeared on the market to improve the process of obtaining bee products. The use of bee products plays a large role in many industries; moreover, the consumption of bee products and promotion of their medicinal properties are very important in shaping proper eating habits.

Effect of in vitro cultivation on human gut microbiota composition using 16S rDNA amplicon sequencing and metabolomics approach.

Gut microbiota (GM) plays many key functions and helps maintain the host's health. Consequently, the development of GM cultivation under in vitro stimulating physiological conditions has gained extreme interest in different fields. In this study, we evaluated the impact of four culture media: Gut Microbiota Medium (GMM), Schaedler Broth (SM), Fermentation Medium (FM), and Carbohydrate Free Basal Medium (CFBM) on preserving the biodiversity and metabolic activity of human GM in batch in vitro cultures using PMA treatment coupled with 16S rDNA sequencing (PMA-seq) and LC-HR-MS/MS untargeted metabolomics supplemented with GC-MS SCFA profiling. Before the experiments, we determined the possibility of using the pooled faecal samples (MIX) from healthy donors (n = 15) as inoculum to reduce the number of variables and ensure the reproducibility of in vitro cultivation tests. Results showed the suitability of pooling faecal samples for in vitro cultivation study. Non-cultured MIX inoculum was characterized by higher α-diversity (Shannon effective count, and Effective microbial richness) compared to inocula from individual donors. After 24 h of cultivation, a significant effect of culture media composition on GM taxonomic and metabolomic profiles was observed. The SM and GMM had the highest α-diversity (Shannon effective count). The highest number of core ASVs (125) shared with non-cultured MIX inoculum and total SCFAs production was observed in the SM. These results might contribute to the development of standardized protocols for human GM in vitro cultivation by preventing methodological bias in the data.