Quantification of aflatoxin M1 in raw milk by a core-shell column on a conventional HPLC with large volume injection and step gradient elution

The immunoaffinity column cleanup method coupled with HPLC separation and fluorescence detection is still one of the most frequently used quantitative methods for routine analyses of AFM1. In this study, aflatoxin M1 (AFM1) was quantified with a chromatography column packed with 2.6 μm core-shell particles on a conventional HPLC. A large volume of solvent (100 μL) was injected into the highly efficient column without any noticeable reduction in separation performance with the help of stepwise gradient elution. The instrumental conditions were optimized by response surface analysis methodology (RSM) with a three-level three-factor Box–Behnken design.The use of core-shell columns on conventional HPLC for AFM1 analysis under the optimized instrumental conditions leads to increased analytical performance compared with traditional totally porous columns without the heavy costs associated ultra-high-pressure instruments. Moreover, the improved instrumental sensitivity enables simplified sample preparation by avoiding any solvent replacement. The method could be easily applied to enhance the sensitivity of HPLC-FLD for AFM1 analysis that is based on isocratic elution and is more widely used. The method was successfully applied to 40 raw milk samples collected in summer from 20 cattle ranches located in two different provinces in southwestern China.     

Detection of aflatoxin M1 in milk products from China by ELISA using monoclonal antibodies

A rapid and sensitive indirect competitive enzyme-linked immunosorbent assay (ELISA) method using monoclonal antibody for measuring aflatoxin M1 (AFM1) in milk and milk products has been described. One monoclonal antibody was isolated and characterized after fusion of myeloma cells with spleen cells isolated from BALB/c mice that had been immunized with AFM1 carboxymethyl oxime conjugated with bovine serum albumin (BSA). Cross-reactivities of the anti-AFM1 monoclonal antibody clone were 100, 13.9, 6.7 and <1% against AFM1, aflatoxin B1 (AFB1), aflatoxin G1 (AFG1) and deoxynivalenol (DON), respectively. Assays of milk samples mixed with AFM1 ranging in concentration from 0.1 to 3.2 ng/ml gave mean ELISA recovery of 98%. The limit of detection concentration of AFM1 was 0.04 ng/ml. AFM1 contamination was measured in 12 samples of raw milk, 15 samples of powdered milk, 104 samples of liquid milk and four cheese samples collected from different supermarkets in Northeast of China. Of 135 milk samples tested, 55 (41%) samples contained AFM1 at levels that ranged from 0.32–0.50 ng/ml, 24 (18%) samples contained 0.16–0.32 ng/ml, and 18 (13%) samples contained 0–0.16 ng/ml; in 38 (28%) samples AFM1 was not detected. The results indicate that the necessary precaution will have to be taken to minimize the AFM1 contamination in milk and milk products from Northeast of China.     

Assessment of aflatoxin M1 levels in pasteurized and UHT milk consumed in Prishtina,Kosovo

To assess public health hazards associated with the occurrence of AFM1 residues in pasteurized milk and UHT milk a survey was carried out, in Prishtina, capital city of Kosovo. In the present study, a total of 178 samples, 84 pasteurized milk and 94 UHT milk were collected during 6 months (July to December 2013). They were obtained from retail outlets in Prishtina city (Kosovo). The occurrence and concentration range of AFM1 in the samples were investigated by competitive enzyme-linked immunosorbent assay (ELISA) method. There was a high incidence of AFM1 (81.0%) in both pasteurized and UHT milk samples. Eighty three percent (83.3%) of the pasteurized milk samples and seventy eight percent (78.7%) of the UHT milk samples contained AFM1. The positive incidence of AFM1 in the pasteurized milk and the UHT milk samples ranged from 5.16 to 110 ng/L and from 5.02 to 62 ng/L, respectively. AFM1 levels in 18 (21.4%) pasteurized milk samples and 4 (4.2%) UHT milk samples exceeded the maximum tolerable limit of the EC according to the European Union regulation limits of 50 ng/L. AFM1 levels in the samples show that there is a presence of high AFM1 level that constitutes a human health risk in Kosovo. The results of this study imply that more emphasis should be given to the routine AFM1 inspection of milk and dairy products in the Prishtina region.     

Seasonal effect on aflatoxin M1 contamination in raw and UHT milk from Croatia

Aflatoxin M1 contamination was examined in raw milk (3716) and UHT milk (706) samples collected from farms of eastern Croatia and markets of central Croatia from February to July 2013. A maximal mean AFM1 of 1135.0 ng/L was measured in raw milk in March. The AFM1 levels exceeded the European Union maximum residue permitted amount (EU MRL) in 45.9% raw and 36.2% UHT milk samples in February. In total, AFM1 levels exceeded the EU MRL values in 27.8% of raw and 9.64% of UHT milk samples. A slight decrease in the number of samples exceeding the EU MRLs was recorded in the period March to June. The results showed significant statistical differences between the mean AFM1 concentrations of raw and UHT milk samples collected during February, March, May and June (P < 0.05, all). Also, statistical differences in AFM1 concentrations were found between months for raw and UHT milk (P < 0.001, both). In conclusion, the frequency of control of feed and milk samples should increase and should strive to educate breeders and those involved in milk production about the harmful effect of aflatoxins to animal feed.     

Occurrence and factors associated with aflatoxin contamination of raw peanuts from Lusaka district's markets,Zambia

Peanuts, one of the most susceptible crops to aflatoxin (AF) contamination, are widely produced and consumed in Zambia. This cross-sectional study was designed to determine the levels of AFs in raw peanuts sold in Lusaka district's markets as well as identify factors associated with increased AF presence. Raw peanut samples were collected from open markets and supermarkets and analyzed for aflatoxin contamination using high performance liquid chromatography (HPLC). A questionnaire was also administered to the peanut vendors to investigate factors contributing to increased levels of AFs in peanuts. Of the 92 samples, 51 (55.4%; 95% CI: 44.9–65.4) tested positive for presence of AFs. The overall median and geometric mean ± standard deviation (SD) concentration for AF were 0.23 ppb (range: 0.014–48.67 ppb) and 0.43 ± 9.77 ppb, respectively. The association between market types and presence of AFs was not statistically significant (Pearson Χ² = 0.0587, p = 0.809). Of 51 samples that tested positive to AF, 6.5% and 12% were above the maximum permissible limits (MPLs) set by the Codex Alimentarius Commission and European Union standards, respectively. There was a significant difference in the levels of AF between Chalimbana and Kadononga (p<0.0001), and also Chalimbana and Makulu red (p<0.0001). Chalimbana was the most at risk of AF contamination, when compared to other peanut varieties. The high level of AFs in raw peanuts from both supermarkets and open markets samples constitutes a health hazard for the population of Lusaka district. Therefore, intervention strategies that reduce the levels of AF contamination in peanuts should be given priority.     

Aflatoxin contamination in unrecorded beers from Kenya – A health risk beyond ethanol

Samples of unrecorded opaque beers (n = 58; 40 based on maize, 5 on sorghum and 13 on other plants) and recorded wines (n = 8) in Kenya were screened for aflatoxins using a rapid ELISA technique followed by confirmation using liquid chromatography-tandem mass spectrometry. Six of the maize beers were obtained from Kibera slums in Nairobi County. Aflatoxin contamination was detected in six unrecorded beers (10%), but in none of the recorded wines. Remarkably, three of the aflatoxin positive samples were from the Kibera slums.The mean concentration of aflatoxins in the positive samples was 3.5 μg/L (range 1.8–6.8 μg/L), corresponding for an average consumption of 500 mL (1 standard drink) to a margin of exposure (MOE) of 36 (range: 15–58), which is considered as ‘risk’. On the other hand, the alcoholic strength of the aflatoxin positive samples had a mean of 4.3% vol (range 3.5–4.8%) corresponding to a MOE of 2.5 (range of 2.2–3.0) for the equivalent consumption volume. While aflatoxins pose a risk to the consumer, this risk is about 10 times lower than the risk of ethanol.The Joint FAO/WHO Expert Committee on Food Additives sets no acceptable daily intake for aflatoxins since they are genotoxic carcinogens and instead recommends for the reduction of aflatoxin dietary exposure as an important public health goal, particularly in populations who consume high levels of any potentially aflatoxins-contaminated food. Nevertheless, ethanol still posed a considerably higher risk in the unrecorded beers examined. However, consumers should be informed about aflatoxins, as these are an involuntary and unknown risk to them. In addition, producers should be educated about measures to reduce aflatoxins in alcoholic beverages.     

Determination of melamine in milk using surface plasma effect of aggregated Au@SiO2 nanoparticles by SERS technique

A simple and rapid method to detect melamine in liquid milk by Surface-enhanced Raman Scattering (SERS) technique was presented. The pretreatment procedure of milk samples only contains hydrochloric acid treatment and twice centrifugation. In order to reduce the distortion about the Raman signals originating from charge transfer and electronic tunnelling effect, SiO₂ shell-isolated gold nanoparticles (Au@SiO₂ NPs) instead of Au NPs were employed to enhance signal intensity. Aggregation occurs when the Au@SiO₂ NPs colloid is mixed with the melamine solution or the treated milk containing melamine. Different from aggregated Au NPs, these aggregated Au@SiO₂ NPs on Cu substrate can undistortedly enhance the Raman signals of melamine using surface plasma effect. Quantitative analysis was tried and the results showed a good linearity (R² ≈ 0.99) when the melamine concentration was between 0.5 mgL⁻¹ and 5 mgL⁻¹. The detection sensitivity satisfies the requirement of the Codex Alimentarius Commission and this method can be practically used for melamine detection in milk.     

Two year survey on the occurrence and seasonal variation of aflatoxin M1 in milk and milk products in Serbia

The incidence of contamination of aflatoxin M1 (AFM1) in milk and milk products samples collected in Serbia was investigated by using the competitive enzyme linked immunosorbent assay (ELISA) technique. A total of 1438 samples composed of 678 raw milk, 438 heat treated milk and 322 milk product samples that were analyzed during the period of 2013–2014, including all seasons. The AFM1 levels exceeded the European Union maximum residue permitted amount (EU MRL) in 56.3% raw milk, 32.6% heat treated milk and 37.8% of milk product samples. Milk powders had the highest mean AFM1 concentration (0.847 μg/kg) of all types of milk products examined. Mean concentration of AFM1 in raw milk samples during the period of winter in Serbia was 0.358 μg/kg and did not significantly differ from the mean concentrations of 0.375 μg/kg during the spring. However, the AFM1 raw milk concentration in the summer (0.039 μg/kg) and autumn season (0.103 μg/kg) was significantly lower. Seasonal variation of AFM1 concentrations in heat treated milk samples followed the trend observed in raw milk. Mean raw milk AFM1 concentration has dropped down by 10 fold from 0.314 μg/kg in 2013 to 0.035 μg/kg in 2014. The fraction of raw milk samples exceeding the EU MRL has decreased from 62.3% to 11.5% by the end of 2014.     

Incidence of aflatoxin M1 in human breast milk in Tehran, Iran

This study examined the exposure of infants to aflatoxin M1 (AFM1) and of lactating mothers to aflatoxin B1 (AFB1), using AFM1 in breast milk as a biomarker for exposure to AFB1. An enzyme-linked immunosorbent assay (ELISA) was modified for the analysis of AFM1 in breast milk samples from 160 women in Tehran, Iran. AFM1 was detected in 157 samples by average concentration of 8.2 ± 5.1 ng/kg (range 0.3–26.7 ng/kg).The concentration of AFM1 in one sample was higher than the maximum tolerance limit accepted by European Union and USA (25 ng/kg), but in 55 samples was higher than the maximum concentration recommended by Australia and Switzerland (10 ng/kg).Logistic regression Analysis failed to show significant correlation between AFM1 and gestational age, education, postnatal age, gender, nationality, clinical condition, the number of family member, the number of children, type and amount of dairy consumption, vegetable, fruits, oil and meat. But it was significant relation to the cereal consumption, also to the height at birth.     

Three years of monitoring of PCDD/F,DL-PCB and NDL-PCB residues in bovine milk from Lombardy and Emilia Romagna regions (Italy): Contamination levels and human exposure assessment

In the three-years period 2012–2014, 160 cow milk samples from farms located in Lombardy and Emilia Romagna regions (Italy) were analyzed during the implementation of the Italian National Residues Monitoring Plan to assess the presence of PCDD/F, DL-PCB and NDL-PCB residues. The obtained contamination data were combined with cow milk consumption data from the Italian national dietary survey to estimate PCDD/F, DL-PCB and NDL-PCB human dietary exposure through the consumption of whole, semi skimmed and skimmed bovine milk. The exposure assessment was carried out separately for children, teenagers, adults and elderly. Average contamination levels of the analyzed samples were found to be 1.26 pg WHO-TEQ/g fat for the sum of PCDD/Fs and DL-PCBs and 9.30 ng/g fat for the sum of the 6 NDL-PCB indicators. PCB 126 was found to be the main contributor to the total WHO-TEQ. Using the upper bound approach, the estimated mean dietary intakes ranged from 0.07 pg WHO-TEQ/kg bw per day to 0.39 pg WHO-TEQ/kg bw per day for the sum of PCDD/Fs and DL-PCBs and considering exposure from whole milk. NDL-PCB mean dietary intakes resulted between 0.52 ng/kg bw per day and 2.86 ng/kg bw per day for consumption of whole milk. Children and teenagers were found to be the most exposed groups. This is the first time that Italian consumers exposure to NDL-PCBs is assessed using contamination data of cow milk produced in Italy.     

Aflatoxin M1 contamination in raw milk from major milk-producing areas of China during four seasons of 2016

This survey was performed to determine the frequency with which raw milk from the major milk-producing areas of China was contaminated with aflatoxin M1 (AFM1) in 2016. In total, 5650 raw milk samples produced during the four seasons of 2016 were collected from the major milk-producing areas of China, including Hebei, Heilongjiang, Henan, Inner Mongolia, Shandong, and Xingjiang provinces. Contamination of AFM1 was detected in 267 of the 5650 raw milk samples in totally, with the incidence of 4.7%. Only 1.1% of raw milk samples exceeded the European Union legal limit (50 ng/L), and none of samples exceeded the Chinese and United States legal limit (500 ng/L). The incidence of AFM1 contamination in raw milk samples was much higher during winter (10.2%) than in spring, summer, or autumn (3.0%, 2.1%, and 4.4%, respectively) in China. Thus, it is particularly important to monitor AFM1 contamination in raw milk during the winter season. This comprehensive study will facilitate future risk analysis and the management of AFM1 contamination in raw milk in China.     

Levels of aflatoxin M1 in different types of milk collected in Serbia: Assessment of human and animal exposure

The level of aflatoxin M1 (AFM1) in 50 milk samples collected from February to June 2013 from Serbian market or domestically produced was determined using simple non-specific sample preparation method based on solid phase extraction (Oasis HLB, Waters) and ultra-high performance liquid chromatography with heated electrospray ionization triple quadrupole mass spectrometry (UHPLC/HESI-MS/MS). The range of detection was between < LOD and 1.44 μg/kg with mean value of 0.30 μg/kg. Thirty-eight samples (76%) exceed the maximum level of 0.05 μg/kg sets by EU. The highest level of 1.44 μg/kg was found in raw sample of domestically produced milk while the lowest one in organic produced milk. The evaluation of the exposure degree of AFM1 through the milk consumption by the average Serbian citizen was estimated at levels of 1.420, 0.769 and 0.503 ng/kg bw/day during February, April and May, respectively. Estimation of the corresponding concentration of AFB1 in feedstuffs was evaluated as 18.75 μg/kg. The calculated hazard index of 7.1, 3.8 and 2.5 for February, April and May, respectively, was higher than 1 indicated serious risk of AFM1 to Serbian consumers. This work presents the first insight in the occurrence of AFM1 in milk collected in Serbia as well as mycotoxin intake through milk consumption by Serbian adult population.     

Occurrence and co-occurrence of Fusarium mycotoxins in wheat grains and wheat flour from Romania

In this study, the presence of fourteen Fusarium mycotoxins, legislated by the European Union – deoxynivalenol, zearalenone, HT-2 and T-2 toxins (EC/1881/2006; 2013/165/EU), or non-legislated (five trichothecens and five “emerging” mycotoxins), was evaluated in 31 whole unprocessed wheat samples and 35 white wheat flour samples from different areas of Romania. For this purpose, a validated multi-mycotoxins liquid chromatography tandem mass spectrometry method was applied. Seventy three percent of the analyzed samples contained at least one mycotoxin. The highest occurrence was for enniatin B, 71% of the analyzed samples being positive (21–407 μg kg⁻¹). Regarding the legislated mycotoxins, deoxynivalenol was detected in 14% (111–1787 μg kg⁻¹) of the samples, while zearalenone was detected in 9% (51–1135 μg kg⁻¹). Only one sample was positive for neosolaniol. Concerning the co-occurrence, 42% of the samples were contaminated with two to five mycotoxins, the most frequent being the binary or tertiary combinations of enniatins. This is the first study applied to Romanian wheat grains and flour samples using a high sensitive multi-mycotoxins method, and which included also “emerging” mycotoxins.     

Rapid multiresidue and multi-class screening for antibiotics and benzimidazoles in feed by ultra high performance liquid chromatography coupled to tandem mass spectrometry

An analytical strategy was developed for high-throughput screening of multiple antibiotics and two benzimidazoles in feed. Generic sample processing was applied without any purification step. After methanol extraction, the samples were centrifuged, concentrated, and analysed by ultra-high-performance liquid chromatography hyphenated to tandem mass spectrometry in the multiple reaction monitoring mode. Qualitative validation was carried out for more than 50 antibacterials of various classes, including aminocoumarin, amphenicols, beta-lactams, lincosamide, macrolides, diaminopyrimidine, quinolones, sulfonamides, streptogramin, pleuromutilin, polypeptide, quinoxaline, and tetracyclines, and also some benzimidazoles in feed at μg/kg level. Validation was done in accordance with the guidelines laid down in European Commission Decision 2002/657/CE for qualitative screening methods.This convenient, reliable, and sensitive method has been used successfully to monitor antibiotic residues in feeds.     

Multiple mycotoxin co-occurrence in maize grown in three agro-ecological zones of Tanzania

In this study, the co-occurrence of multiple mycotoxins in maize kernels collected from 300 households' stores in three agro-ecological zones in Tanzania was evaluated by using ultra high performance liquid chromatography/time-of-flight mass spectrometry (TOFMS) with a QuEChERS-based procedure as sample treatment. This method was validated for the analysis of the main eleven mycotoxins of health concern that can occur in maize: aflatoxin B₁ (AFB₁), aflatoxin B₂ (AFB₂), aflatoxin G₁ (AFG₁), aflatoxin G₂ (AFG₂), ochratoxin A (OTA), deoxynivalenol (DON), fumonisin B₁ (FB₁), fumonisin B₂ (FB₂), HT-2 toxin, T-2 toxin and zearalenone (ZEN). From each zone one major maize producing district for home consumption was chosen and 20 villages for each district were randomly selected for sampling. All mycotoxins of health concern, except for T-2 toxin, were detected in the maize samples. Particularly high levels of AFB₁ (50%; 3–1,081 μg kg⁻¹), FB₁ (73%; 16–18,184 μg kg⁻¹), FB₂ (48%; 178–38,217 μg kg⁻¹) and DON (63%; 68–2,196 μg kg⁻¹) were observed. Some samples exceeded the maximum limits set in Tanzania for aflatoxins or in European regulations for other mycotoxins in unprocessed maize. Eighty seven percent of samples were contaminated with more than one mycotoxin, with 45% of samples co-contaminated by carcinogenic mycotoxins, aflatoxins and fumonisins. Significant differences in contamination pattern were observed among the three agro-ecological zones. The high incidence and at high levels (for some) of these mycotoxins in maize may have serious implications on the health of the consumers since maize constitute the staple food of most Tanzanian population. Effective strategies targeting more than one mycotoxin are encouraged to reduce contamination of maize with mycotoxins.     

Screening of aflatoxin M1 occurrence in selected milk and dairy products in Terengganu,Malaysia

The study was conducted to screen the occurrence of aflatoxin M₁ (AFM₁) in 53 selected milk and dairy product samples (11 liquid milk, 12 powdered milk, 8 3-in-1 beverages, 6 condensed sweetened milk, 2 evaporated milk, 7 cultured milk drink, 5 yogurt and 2 cheese samples). These samples were purchased from selected markets in Terengganu, Malaysia in January 2014 based on a questionnaire survey among 212 respondents on the types and brands of milk and dairy products that were frequently consumed. Based on the responses, 53 milk and dairy products were purchased and the competitive enzyme-linked immune-absorbent assay (ELISA) method was used to determine the level of AFM₁ in the samples. Of 53 samples, 19 samples were positive with AFM₁ (35.8%) ranging from 3.5 to 100.5 ng/L. Although 4/53 (7.5%) of the tested samples had the contamination level greater than the European Commission (EC) limit (>50 ng/L), the contamination levels were still below the Malaysia Food Regulation 1985 limit (less than 500 ng/L). This study provided a pioneering data on the occurrence of AFM₁ in milk and dairy products in Malaysia.     

Verifying the red wines adulteration through isotopic and chromatographic investigations coupled with multivariate statistic interpretation of the data

The assessment of wine traceability and authenticity is a major concern that has gained a lot of interest internationally since the wine has always been subjected to various fraudulent practices. Practiced since ancient times, wine fraud has become more sophisticated in the present day, taking many forms. Consumers, regulatory bodies and manufacturers are all interested to have reliable analytical tools and information to allow the authentication and detection of wine adulteration or incorrect labelling. This research study evaluates and proposes a possible strategy for the detection of adulterated sweet or medium sweet red table wines using appropriate chemical parameters which can reveal prohibited practices in the winemaking process. The work is performed on 29 table wine samples, bought from the market, packed in PET bottles. Exogenous addition of sugar and water in the counterfeited table red wines was detected by the measurement of stable isotopes content (δ¹³C and δ¹⁸O) known as origin markers and supplementary confirmed by other classical parameters, as the alcoholic strength of the wines (‰ vol.) and the presence of 5-(hydroxymethyl)-2-furaldehyde (HMF) and of synthetic sweeteners or synthetic red dyes used to correct deficiencies of taste and colour. Additionally, the nature and profile of anthocyanins, as indicator of the red colour of wine, was investigated in the table wines and then compared with that of authentic wines obtained by microvinification of Vitis vinifera, in order to determine their authenticity.     

Latex bead and colloidal gold applied in a multiplex immunochromatographic assay for high-throughput detection of three classes of antibiotic residues in milk

Two rapid and sensitive high-throughput immunochromatographic assays were successfully developed for simultaneous determination of 12 sulfonamide, 18 quinolone and six tetracycline residues in milk. Under optimized conditions, residues from these three classes of antibiotics can be qualitatively and quantitatively determined on a single strip within 10 min. The detection limits were much lower than the maximum residue limits established by the European Union. Comparative evaluations of sensitivity, antibody consumption, coefficient of variation and user experience between these two immunochromatographic assays showed that latex beads have advantages over colloidal gold when employed as a label in the assay. Therefore, the latex bead-based high-throughput immunochromatographic assay has the best potential for on-site testing of a large number of samples as part of food safety applications.     

Assessment of aflatoxin M1 contamination in the milk of four dairy species in Croatia

Aflatoxin M1 (AFM1) concentrations were measured in bulk cow milk samples from eastern Croatia, and in cow, goat, sheep and donkey bulk milk samples from other parts of Croatia during the period July–September 2013. AFM1 levels in milk were measured in the ranges (ng/L): cow 3.65–162.3 (eastern Croatia) and 2.69–44.9 (other regions of Croatia); goat 2.78–40.8; sheep 2.11–5.87; donkey 3.43–10.4. The concentration of AM1 exceeded the EU MRL in 6.7% of cow milk samples from eastern Croatia. The highest level measured was 162.3 ng/L. AFM1 levels exceeded the LOQ value (23.2 ng/L) in only 59 samples of cow milk and two samples of goat milk of the total 402 samples analysed. A significant difference was found between the mean AFM1 concentrations of cow milk from eastern and other regions of Croatia (P < 0.05). The elevated AFM1 levels in cow milk from eastern Croatia indicate the use of contaminated supplementary feedstuff in some farms during the study period.     

Inhibition of Fusarium culmorum by carboxylic acids released from lactic acid bacteria in a barley malt substrate

The effect of carboxylic acids, composed by both organic and phenolic acids, released in a barley malt substrate fermented by lactic acid bacteria was tested against Fusarium culmorum macroconidia and compared under different fermentation conditions. Phenolic acids released by Lactobacillus plantarum FST1.7 and Lactobacillus brevis R2Δ were quantified using a QuEChERS method coupled with a HPLC-UV/PDA system. Their concentration improved with increasing extract content of the barley malt-based substrate and reached maximal concentrations after 48 h of fermentation performed at optimum growth temperature. Generally, phenolic acids were produced at levels far below their minimal inhibitory concentration (MIC), and limited synergistic effects were observed when mixed with organic acids. The fungal growth suppression by the wort fermented by Lb. brevis R2Δ (95 ± 9 h total inhibition) could be fully explained by the presence of antifungal carboxylic acids, whereas only partially accounted for Lb. plantarum FST1.7 (198 ± 19 h). Organic acids were mainly responsible for the ability of LAB fermented wort to cause fungal inhibition, whereas phenolic acids took only a secondary role at the low concentrations released. Longer fermentation times favoured primarily organic acid release, whereas fermentation of higher malt extract substrates encouraged both organic and phenolic acids production. The understanding on how synergy works between antifungal compounds could help to identify strategies to further increase their concentration in wort, with potential to replace synthetic broths and for direct application in food application.