Presence Of Mycotoxins Research Articles

Development of a Microwave-Assisted Extraction Protocol for the Simultaneous Determination of Mycotoxins and Pesticide Residues in Apples by LC-MS/MS

The possible presence of contaminants, pesticide residues and mycotoxins, in agricultural commodities is a critical issue for food safety, causing great concern. In this work, a simple and rapid analytical method employing liquid chromatography–tandem mass spectrometry (LC-MS/MS) was developed for the simultaneous determination of pesticide residues and mycotoxins in apples. Microwave-assisted extraction (MAE) was used for sample preparation. The MAE protocol was optimized after evaluating the effects of the following extraction parameters: (a) extraction solvent, (b) acidic environment, (c) temperature, and (d) extraction time. The multiresidue MAE-LC-MS/MS method was validated for linearity, accuracy (bias and precision), limits of detection (LODs), limits of quantification (LOQs), and matrix effect. The validation of the method was carried out according to the SANTE/12682/2019 document. The method demonstrated good linearity with R2 ≥ 0.99, acceptable accuracy in the recovery rate range 70–116%, acceptable interassay precision with RSD% ≤ 20, and low LODs and LOQs in the ranges 0.005–0.015 μg/g and 0.01–0.03 μg/g, respectively. Matrix effects were observed only for the 25% of the analytes. The performance of the MAE-LC-MS/MS method was compared to that of the QuEChERS sample preparation method, and the MAE-LC-MS/MS method proved to be rapid and effective.

Species of the genus Fusarium and Fusarium toxins in the grain of winter and spring wheat in Poland

The aim of the study was to determine the presence of Fusarium species and mycotoxins in wheat grain from harvest in 2009 and 2010 in Poland. Samples from different locations were analyzed for the content of DNA of Fusarium species and mycotoxins. In 2009, DNA of F. graminearum and F. poae was present in all samples, F. culmorum in 82% of samples, and F. avenaceum in 55% of samples. In 2010, the highest content of DNA was found for F. graminearum followed by F. avenaceum, F. poae and F. langsethiae. The amount of F. culmorum DNA was very low. The most frequently occurring species were F. poae and F. graminearum, however, the amount of F. poae DNA was lower. In 2009, deoxynivalenol was detected in all samples. In 2010, the average content of deoxynivalenol was lower than in 2009. Nivalenol was detected at very low concentration in both years. Significant correlations between content of F. graminearum DNA and deoxynivalenol concentration in the grain and between content of F. poae DNA and nivalenol concentration in the grain in 2009 were found.

Mycotoxins—Prevention, Detection, Impact on Animal Health

Mycotoxins are defined as secondary metabolites of some species of mold fungi. They are present in many foods consumed by animals. Moreover, they most often contaminate products of plant and animal origin. Fungi of genera Fusarium, Aspergillus, and Penicillum are most often responsible for the production of mycotoxins. They release toxic compounds that, when properly accumulated, can affect many aspects of breeding, such as reproduction and immunity, as well as the overall liver detoxification performance of animals. Mycotoxins, which are chemical compounds, are extremely difficult to remove due to their natural resistance to mechanical, thermal, and chemical factors. Modern methods of analysis allow the detection of the presence of mycotoxins and determine the level of contamination with them, both in raw materials and in foods. Various food processes that can affect mycotoxins include cleaning, grinding, brewing, cooking, baking, frying, flaking, and extrusion. Most feeding processes have a variable effect on mycotoxins, with those that use high temperatures having the greatest influence. Unfortunately, all these processes significantly reduce mycotoxin amounts, but they do not completely eliminate them. This article presents the risks associated with the presence of mycotoxins in foods and the methods of their detection and prevention.

Effect of a Mycotoxin Binder (MMDA) on the Growth Performance, Blood and Carcass Characteristics of Broilers Fed Ochratoxin A and T-2 Mycotoxin Contaminated Diets.

Simple SummaryThe contamination of feed with mycotoxins is a global concern, resulting in adverse effects on productivity and animal health and, therefore, a great economic loss. Ochratoxin A and T-2 mycotoxins are among the mycotoxins that contaminate animal feed. These mycotoxins could adversely affect the health of broilers, and the most effective method to mitigate the toxic effects of mycotoxins is the use of detoxifying agents. In the present experiment, broiler chickens were allotted into five groups. Group 1 received a non-contaminated diet; group 2 received a non-contaminated diet + 3 g/kg of a mycotoxin binder (MMDA); group 3 received a non-contaminated diet + 0.5 mg/kg OTA + 1 mg/kg T-2 toxin; group 4 received a non-contaminated diet + 0.5 mg/kg OTA + 1 mg/kg T-2 toxin + 1 g/kg MMDA; and group 5 received a non-contaminated diet + 0.5 mg/kg OTA + 1 mg/kg T-2 toxin + 3 g/kg MMDA for 35 days. The results revealed that OTA and T-2 toxin negatively affected the productive parameters and some blood and carcass characteristics of broiler chickens. The addition of the detoxifying agent (MMDA at 1 or 3 g/kg feed) to contaminated diets alleviated the adverse effects observed on productivity and the broilers heath related parameters. The present study was conducted to evaluate the efficacy of the feed additive, a novel multicomponent mycotoxin detoxifying agent (MMDA) containing modified zeolite (clinoptilolite), Bacillus subtilis, B. licheniformis, Saccharomyces cerevisiae cell walls, and silymarin, as detoxifiers of 0.5 mg/kg (0.5 ppm) ochratoxin A (OTA) and 1 mg/kg (1 ppm) T-2 toxin on broiler chickens. A total of 240 1-old broiler chickens (Ross 308) were randomly distributed into five different dietary treatments: (1) control (non-contaminated diet); (2) non contaminated diet + 3 g/kg of MMDA; (3) non-contaminated diet + 0.5 mg/kg OTA + 1 mg/kg T-2 toxin; (4) non-contaminated diet + 0.5 mg/kg OTA + 1 mg/kg T-2 toxin + 1 g/kg MMDA; and (5) non-contaminated diet + 0.5 mg/kg OTA + 1 g/kg T-2 toxin + 3 g/kg MMDA. The results showed that, in the starter period, from 1 to 10 days, the presence of OTA and T-2 mycotoxins reduced the consumption of feed and the growth of the broilers, and no effects of the detoxifying product were observed in the productivity of the chickens, at any of the doses tested, compared to the contaminated control (treatment 3). However, in the growing period, the same negative effect of mycotoxins was registered, but a recovery was observed in the consumption of feed and in the weight of the broilers that consumed 3 g/kg of the MMDA mycotoxin binder, reaching similar values to those of chickens fed uncontaminated control diets. The presence of mycotoxins in feed led to a reduction in the concentration of total proteins and albumin in blood compared to controls, and the presence of the detoxifying product partially reversed this effect. The breast yield of the chickens fed with mycotoxins was lower than that of the animals fed with the control feed and was not affected by the presence of the product tested, at 1 or 3 g/kg. The weight of the different organs (liver, gizzard, kidneys, or spleen), the intestinal pH, the histology of the small intestine, and oral lesions were not affected by the experimental treatments. In summary, the productive parameters and some blood and carcass characteristics of broiler chickens were impaired by the dietary presence of OTA and T-2 toxin. The tested product included at 1 or 3 g/kg feed in contaminated diets improved performance and seems to be effective in partly counteracting the deleterious effects of the tested mycotoxins.

Investigation of an acute neurological outbreak in Eluru, India, 2020.

On 4th December 2020, a sudden outbreak, with neurological symptoms like seizures, loss of consciousness etc., was reported in a town from south India. By 3rd day about 400 people were involved. A multi disciplinary team from our institute visited the site to investigate the outbreak. Based on the case history and clinical examination of the patients, the team suspected a probable diagnosis of an acute pesticide, heavy metal and/or mycotoxin exposure for which, biological samples (blood, urine) were collected from those who reported the symptoms as well as from a few who did not report symptoms (controls). To identify the source, water and food samples were collected. The samples were subjected to ICP-MS for heavy metal analysis, LC-MS/MS for pesticide analysis, microbiological analysis and ELISA-Kit method for aflatoxins if any. Clinical and dietary details were collected from a total of 112 participants, of which, 103 cases (77 active cases at Hospital and 26 recovered cases from community) and 9 were controls. A total of 109 biological samples, 36 water samples and food samples were collected. The mean age of the study participants was 29.2 years. Among cases, Seizures were seen in 84%, loss of consciousness in 66%, mental confusion in 35%, pinpoint pupil in 11%. Triazophos (organophosphate) pesticide was present in 74% of Blood samples and its metabolites were present in 98% of the urine samples collected from the cases. All the ten heavy metals investigated including lead, mercury and nickel were found to be within permissible limits except for a few samples. No presence of mycotoxins was observed in Food samples. Water samples which included Head pump and reservoir were free from pesticides; however, all water samples from households of cases had triazophos pesticide with a mean concentration of 1.00 ug/L. Thus, it was concluded that, the probable cause of outbreak was Triazophos (Organophosphate) pesticide contamination in water at the Household level. Regular surveillance for the presence of residual pesticides in soil, water and food with heightened vigour is recommended to prevent future outbreaks.

Evaluating the effects of cellulolytic enzymes and Lactobacillus bulgaricus on mycotoxins production and the quality of maize silage

Fungal spoilage and mycotoxin contamination are two of the greatest hazards of silage. The present work was carried out to evaluate the impact of Lactobacillus bulgaricus and cellulolytic enzymes on the maize silage (MS) quality. Fungal analysis of different MS samples showed different mycotoxigenic fungi. The highest frequency (62.8%) was associated with Fusarium spp. Four species with different relative densities were found: F. graminearum (71.1%), F. culmorum (15.2%), F. proliferatum (11.2%), and F. oxysporum (2.50 %). High-performance liquid chromatography analysis showed the presence of trichothecene, nivalenol, zearalenone, and fumonisins mycotoxins in MS inoculated by F. graminearum. The inhibition % of trichothecene, nivalenol, and zearalenone synthesis was 50.2%, 47.5%, and 23.5%, respectively, in MS inoculated by Lactobacillus bulgaricus after a 30 d incubation period. Trichoderma harzianum succeeded in producing cellulolytic enzymes, i.e., carboxymethyl cellulase, manganase peroxidase, and laccase, with a maximum production of 350 µg/mL, 5.47 µg/mL, and 16.0 µg/mL, respectively, after 21 d using MS as the substrate. Treatment by the extracted cellulolytic enzyme with L. bulgaricus enhanced unfavorable conditions for MS fungal contamination, i.e., the production of lactic acid, a lowered pH, and increased L. bulgaricus colony-forming units, compared to the addition of enzyme extract or L. bulgaricus alone.

A novel aptasensor based on DNA hydrogel for sensitive visual detection of ochratoxin A.

Anovel visual detection mode isproposed to improve the detection sensitivityfor the determination of ochratoxin A (OTA). The mode is based on aptamer recognition and the signal amplification of rolling circle amplification (RCA) products self-assembled DNA hydrogel. Moreover, gold nanoparticles (AuNPs) were directly assembled inside the DNA hydrogel by adjusting the padlock probe sequences to achieve a stronger binding force between the DNA hydrogel and AuNPs; this avoids the need for modification of AuNPs with DNA sequences. In the presence of OTA, DNA hydrogel is formed. With higher concentrations of OTA, a larger amount of DNA hydrogel is formed. When AuNPs are added to the DNA hydrogel, AuNPs can be enclosed inside the DNA hydrogel. With more DNA hydrogel, there is less AuNPs in the supernatant. Thus, the absorbance of the supernatant is anti-correlated with the concentration of OTA. After optimization of the experimental conditions, the change in the absorbance of the supernatant was linearly correlated with the concentration of OTA, in the range 0.05 to 10ng/mL; the limit of detection was 0.005ng/mL. The good specificity of the developed biosensor was confirmed in the presence of other mycotoxins that are coexistent withor analogues of OTA. By comparing the developed method with the ELISA method, the accuracy and stability of this new method were also verified, with good performance obtained in real samples. Diagram of the principle of the colorimetric aptasensor for OTA detection based on rolling circle amplification product self-assembled DNA hydrogel.

Fusarium head blight in the Russian Far East: 140 years after description of the 'drunken bread' problem.

The first appearance of Fusarium head blight (FHB)—and the beginning of scientific research of this disease—occurred the Far East region of Russia at the end of the 19th century. In the summer of 2019, in the Amur region, which comprises 60–70% of grain production in the Russian Far East, flooding caused a state of emergency. The quality of wheat and barley grains grown under natural conditions of FHB outbreaks, including grain infection, fungal species composition, DNA content of F. graminearum and chemotypes, and the presence of various mycotoxins, was studied. Fusarium infection rates reached extremely high percentages, 51–98%, the majority of which were F. graminearum infections. The amount of F. graminearum DNA in wheat grain samples was higher than in the barley grain samples and averaged 6.1 and 2.1 pg/ng, respectively. The content of deoxynivalenol (DON) in the wheat samples reached 13,343 ppb and in barley reached 7,755 ppb. A multilocus genotyping assay was conducted on the partially sequenced fragments of the translation elongation factor EF-1a, ammonium ligase gene, reductase gene, and 3-O-acetyltransferase gene in 29 Fusarium graminearum sensu lato strains from the grain harvested in the Amur region. All strains from the Far East region were characterized as F. graminearum sensu stricto; 70% were the 15-AcDON chemotype, while the other strains were the 3-AcDON chemotype. According to the results, after 140 years of study of FHB, we are still not very successful in controlling this disease if conditions are favorable for pathogen development. Even at present, some of the grain harvested must be destroyed, as high contamination of mycotoxins renders it unusable.

Examination of wheat kernels for the presence of Fusarium damage and mycotoxins using near-infrared hyperspectral imaging

The agriculture industry experiences severe economic losses each year due to infection of cereal crops with Fusarium, often leading to contamination with the secondary metabolite mycotoxin Deoxynivalenol (DON). Timely detection of Fusarium Damaged Kernels (FDK) could provide producers with an opportunity to develop and implement mitigation strategies and minimize corresponding losses, and also to avoid health risks to humans and animals. Given the slow, labor-intensive, and often destructive current FDK and DON detection methods, the agriculture industry is in dire need of fast, non-invasive, and non-destructive alternative screening techniques. Recent research efforts have introduced hyperspectral imaging (HSI) as a promising non-destructive tool for FDK and DON risk management in cereals. However, a simple single scan for both FDK and DON identification is highly desired due to its ease of use and resulting commercial acceptance. To this end, the present research investigated the feasibility of using NIR HSI in the 960–1700 nm wavelength range to detect the presence of FDK and its secondary metabolite DON in Canadian Western Red Spring (CWRS) wheat. The two k-nearest neighbour (kNN)-based classifiers were developed by identifying 0.2% of the classified spectra that identify their classes. These were identified through their consistent categorization using faster 1NN classifiers from samples of spectra. The resulting 3NN classifiers, one for identifying the presence of FDK and the other for determining the presence of DON, gave good results in classification. The FDK classifier obtained 85% accuracy and 92% sensitivity, and the DON classifier achieved 80% accuracy and 77% sensitivity. Our results indicate that a single HSI scan can be used to rapidly identify the presence of both FDK and DON in CWRS wheat samples. The developed model can be used as a pre-screening technique by the wheat industry to reduce the time and labor currently expended in FDK and DON identification.

Risk assessment of multiple-mycotoxin exposure for consumers of chestnuts in Shandong Province markets in China

ABSTRACT In this study, 321 chestnut samples from Shandong Province in China were analysed for the presence of mycotoxins. We screened for 14 mycotoxins including aflatoxins (AFs: AFB1, AFB2, AFG1, AFG2), deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), T-2 toxin (T-2), zearalenone (ZEA), ochratoxin A (OTA), fumonisins (FB1, FB2, FB3), and penicillic acid (PeA). Mycotoxins were detected in 56.4% of the samples, and 11 of these mycotoxins were found. Thirty samples from the Shandong Province markets were deemed positive for AFs (9.3%) and had an AFB1 level of >2 μg/kg or a sum of AFB1, AFB2, AFG1, and AFG2 that was >4 μg/kg, which exceed the maximum tolerable level of the European regulations standards (EC/188/2006). The contamination level for total mycotoxins found in chestnuts was in the range of 0.6–2,791.0 μg/kg. The estimated daily intake (EDI) values for each individual mycotoxin and for all of the mycotoxins collectively were calculated by both a deterministic approach and a probabilistic approach. For risk characterisation, dietary exposure to DON, ZEA, FBs, and OTA through consumption of chestnuts, analysed according to both approaches, showed no health risk to Chinese adolescents and adults from exposure to either individual mycotoxins or in combination, but more concern should be paid to the AFs for adolescents and adults at a high consumption level. This is believed to be the first work performing risk assessment of multiple mycotoxins specifically for adolescents, including the recently isolated FBs and PeA, which have recently emerged as mycotoxins of concern, in chestnuts of Shandong Province in China.

The role of pumpkin pulp extract carotenoids against mycotoxin damage in the blood brain barrier in vitro.

Some mycotoxins such as beauvericin (BEA), ochratoxin A (OTA), and zearalenone (ZEA) can cross the blood brain barrier, which is why we tested the anti-inflammatory action of a pumpkin carotenoid extract (from the pulp) against these mycotoxins and their combinations (OTA+ZEA and OTA+ZEA+BEA) on a blood brain barrier model with co-cultured ECV304 and C6 cells using an untargeted metabolomic approach. The cells were added with mycotoxins at a concentration of 100 nmol/L per mycotoxin and pumpkin carotenoid extract at 500 nmol/L. For control we used only vehicle solvent (cell control) or vehicle solvent with pumpkin extract (extract control). After two hours of exposure, samples were analysed with HPLC-ESI-QTOF-MS. Metabolites were identified against the Metlin database. The proinflammatory arachidonic acid metabolite eoxin (14,15-LTE4) showed lower abundance in ZEA and BEA+OTA+ZEA-treated cultures that also received the pumpkin extract than in cultures that were not treated with the extract. Another marker of inflammation, prostaglandin D2-glycerol ester, was only found in cultures treated with OTA+ZEA and BEA+OTA+ZEA but not in the ones that were also treated with the pumpkin extract. Furthermore, the concentration of the pumpkin extract metabolite dihydromorelloflavone significantly decreased in the presence of mycotoxins. In conclusion, the pumpkin extract showed protective activity against cellular inflammation triggered by mycotoxins thanks to the properties pertinent to flavonoids contained in the pulp.

Decision support system for integrated management of mycotoxins in feed and food supply chains

Mycotoxins present a global food safety threat of our feed and food. Mycotoxins are toxic metabolites of certain fungi in agricultural products that are harmful to animal and human health. The presence of mycotoxins in these products depends on a variety of management and environmental factors in the field, during storage and/or processing of feed and food commodities. To date, information on mycotoxin management is available, but is not easy to access by supply chain actors. This study aimed to design, build and test a Decision Support System (DSS) that can help decision making on mycotoxin management by various actors along the feed and food supply chains. As part of this, available knowledge and data on mycotoxin prevention and control were collected and synthesised into easy to understand guidelines and tools for various groups of end-users. The DSS consists of four different modules: (a) static information module and (b) scenario analysis module, (c) dynamic module for forecasting mycotoxins, and (d) dynamic module for real-time monitoring of moulds/mycotoxins in grain silos. Intended end-users are all end-user groups for modules (a) and (b); growers and collectors for module (c) and; post-harvest storage managers for module (d). The DSS is user-friendly and accessible through PCs, tablets and smartphones (seehttps://mytoolbox-platform.com/). In various phases of the DSS development, the tool has been demonstrated to groups of end-users, and their suggestions have been taken into account, whenever possible. Also, a near final version has been tested with individual farmers on the easiness to use the system. In this way we aimed to maximise the DSS uptake by actors along the chain. Ultimately, this DSS will improve decision making on mycotoxin management; it will assist in reducing mycotoxin contamination in the key crops of Europe, thereby reducing economic losses and improving animal and human health.

Characterization and Screening of Lactic Acid Bacteria (LAB) and Yeast from Fermented Sorghum and Maize Products for use as Starter Culture

This study focused on the isolation, Characterization and screening of lactic acid bacteria and yeast for use as starter culture in order to eradicate the problem of excessive microbial contamination and the presence of mycotoxins.
 Lactobacillus plantarum and Saccharomyces cerevisiae were the predominant microorganisms isolated from the samples collected. The isolates were screened for their ability to produce enzymes and metabolites. Lactic acid bacteria produce a variety of antimicrobial compounds such as lactic acid, hydrogen peroxide, diacetyl on them as a natural competitive means to overcome other microorganism sharing the same niche. They were also screened for their ability to withstand some physiological stress like acid tolerance, temperature, salt concentration and antibacterial activity; the isolates produced significant values of enzymes and, antimicrobial metabolites. The antimicrobial activity of Lactobacillus plantarum is mainly attributed to the low pH due to Lactic acid production. The antimicrobial activity of the Saccharomyces cerevisiae appears to be mainly due to the competition with the other microorganisms. In addition, depletion of Oxygen and production of CO2, competition for nutrients and the production of antimicrobial substances could have been responsible for the overall antimicrobial activity of both cultures. According to the results obtained, both lactic acid bacteria and yeast proved to be a good source of starter culture.

Mycotoxigenic and phylogenetic perspective to the yeasts and filamentous moulds in mould-matured Turkish cheese

This study was conducted to determine the diversity of yeasts and filamentous moulds in mould-matured cheese (MMC) consumed in Turkey. Overall, 120 samples were collected from 12 different geographical locations between March 2016 and April 2017. The morphological observation was applied in combination with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and molecular analyses to determine yeasts and filamentous moulds in the cheeses. High-performance liquid chromatography (HPLC) technique was used to evaluate the ability of mycotoxins production of fungal isolates and the presence of mycotoxins in cheese samples. A total of 241 fungi (81 filamentous moulds and 160 yeast) were recovered, and Penicillium roqueforti and Debaryomyces hansenii were the most frequently isolated species in all cheese samples. The rep-PCR results indicated a high level of genetic diversity among fungal isolates, regardless of isolation source or geographical origin. Filamentous mould strains isolated from MMC were found to synthesize at least one mycotoxin (Aflatoxin B1, B2, G1 and G2, citrinine, cyclopiazonic acid, mycophenolic acid, ochratoxin A, penicillic acid and roquefortine C). Although mycotoxin producing ability was observed from all isolates, none of the cheese samples were found positive for these mycotoxins. AFM1 was detected in 8 (6.6%) MMC samples from which 2 (1.6%) were above the legal limits (0.05 μg/kg) set by the Turkish Food Codex (TFC) and European Commission (EC). In conclusion, Turkish MMCs were found to be contaminated with toxigenic fungi, so a potential public health risk, while low, exists. Therefore, the selection of nontoxigenic filamentous mould strains for cheese manufacturing and control of the ripening conditions is a critical need to ensure the quality and safety of Turkish MMC.

Fish consumption and consumer awareness aspects of possible mycotoxin contamination in fish in Curitiba-PR, Southern Brazil

The global per capita consumption of fish reached a record of 20.5 kg in 2018, thus demonstrating the sector’s importance. A potential risk to consider when consuming fish is the indirect intake of mycotoxins by humans when consuming fish from intensive fish farming fed with potentially contaminated mycotoxins. The aim of this study was to evaluate fish consumption and consumer awareness of possible mycotoxin contamination in Curitiba, Paraná, Brazil. The information was obtained through a questionnaire applied at supermarkets in 5 different city regions, obtaining 358 respondents. Descriptive analysis was performed on the data, followed by Spearman’s correlation analysis between the responses. It was observed that demographic data (e.g., age, gender, and social class) significantly influenced fish consumption frequency. Thus, 64.80% of respondents preferred tilapia meat over other fish species, 89.91% of people are unaware of mycotoxins, and 93.95% of people do not know what damage mycotoxins cause in human and animal health, and 86.17% of people did not know about any disease related to fish consumption. Schooling had a significant correlation with the aforementioned issues, demonstrating that lower educational levels negatively influence the perception of illnesses caused by food. The most consumed fish is farm-raised tilapia and the consequent possibility of mycotoxin exposure. Respondents are unaware of the possible mycotoxin presence infish and their impact on human health.

Microbiological Safety and Presence of Major Mycotoxins in Animal Feed for Laboratory Animals in a Developing Country: The Case of Costa Rica.

Simple SummaryThe microbiological safety and quality of commercial animal feed for laboratory animals, produced in Costa Rica, was assessed. Analysis of the animal feed included general microbial markers (total coliforms and molds) and the behavior over time of two specific feed contaminants (Salmonella spp. and mycotoxins). Results from the study suggest that there is a low risk of contamination from viable microorganisms but the product contains important levels of mycotoxins. Current preventive measures (UV light disinfection) are not effective and additional handling protocols should be considered.Safety and quality of compound feed for experimental animals in Costa Rica is unknown. Some contaminants, such as Salmonella spp. and mycotoxins, could elicit confounding effects in laboratory animals used for biomedical research. In this study, different batches of extruded animal feed, intended for laboratory rodents in Costa Rica, were analyzed to determine mycotoxin and microbiological contamination (i.e., Salmonella spp., Escherichia coli, total coliform bacteria, and total yeast and molds enumeration). Two methods for Salmonella decontamination (UV light and thermal treatment) were assessed. Only n = 2 of the samples were negative (representing 12.50%) for the 26 mycotoxins tested. Enniatins and fumonisins were among the most frequent toxins found (with n = 4+ hits), but the level of contamination and the type of mycotoxins depended on the supplier. None of the indicator microorganisms, nor Salmonella, were found in any of the tested batches, and no mold contamination, nor Salmonella growth, occurs during storage (i.e., 2–6 months under laboratory conditions). However, mycotoxins, such as enniatins and fumonisins tend to decrease after the fourth month of storage, and Salmonella exhibited a lifespan of 64 days at 17 °C even in the presence of UV light. The D-values for Salmonella were between 65.58 ± 2.95 (65 °C) and 6.21 ± 0.11 (80 °C) min, and the thermal destruction time (z-value) was calculated at 15.62 °C. Results from this study suggest that laboratory rodents may be at risk of contamination from animal feed that could significantly affect the outcomes of biomedical experiments. Thus, improved quality controls and handling protocols for the product are suggested.

Revalorization of Almond By-Products for the Design of Novel Functional Foods: An Updated Review.

The search for waste minimization and the valorization of by-products are key to good management and improved sustainability in the food industry. The great production of almonds, based on their high nutritional value as food, especially almond kernels, generates tons of waste yearly. The remaining parts (skin, shell, hulls, etc.) are still little explored, even though they have been used as fuel by burning or as livestock feed. The interest in these by-products has been increasing, as they possess beneficial properties, caused by the presence of different bioactive compounds, and can be used as promising sources of new ingredients for the food, cosmetic and pharmaceutical industry. Additionally, the use of almond by-products is being increasingly applied for the fortification of already-existing food products, but there are some limitations, including the presence of allergens and mycotoxins that harden their applicability. This review focuses on the extraction technologies applied to the valorization of almond by-products for the development of new value-added products that would contribute to the reduction of environmental impact and an improvement in the sustainability and competitiveness of the almond industry.

Occurrence and concentration of mycotoxins in maize dried distillers’ grains produced in Brazil

The presence of mycotoxins in dried distillers’ grains with solubles (DDGS), a by-product of bioethanol production from maize, has been a matter of concern due to the increasing global utilisation of this ingredient in animal feed. In this study, 186 samples of maize DDGS produced in Brazil were analysed for the presence of major mycotoxins: aflatoxins (B1, B2, G1, and G2), fumonisins (B1 and B2), zearalenone (ZEN), deoxynivalenol (DON) and ochratoxin A (OTA). Samples were provided by the local industry between January 2017 and October 2020, and mycotoxins were quantified by LC-MS/MS. More than 98% of the analysed samples were contaminated with mycotoxins, from which 59.9% had a single mycotoxin, 29.9% two mycotoxins, and 9.1% more than two mycotoxins. The most prevalent metabolites were fumonisin B1 and B2, being detected in 98.8% (mean 3,207 μg/kg) and 97.6% (mean 1,243 μg/kg) of the samples, respectively; aflatoxin B1 had the third highest positivity, with 32.3% (mean 1.47 μg/kg), followed by ZEN, with 18.01% (mean 18.2 μg/kg), DON, with 12.9% (mean 59.6 μg/kg), and OTA was not detected. Co-occurrence of total aflatoxins (AFT = aflatoxin B1+B2+G1+G2) and total fumonisins (FBT = fumonisin B1+B2) was observed in 32.07% of the samples analysed for these mycotoxins. Co-occurrence of AFT and ZEN was found in 7.84% of the samples analysed for such mycotoxins, while FBT and DON co-occurred at 13.01%. AFT, FBT, DON and ZEN co-occurred in only one sample (0.84%). Except for FBT, a considerable number of samples presented the evaluated mycotoxins below their respective limit of quantification (LOQ) with percentages of 67.61% for AFT, 81.99% for ZEN, 87.07% for DON and 100% for OTA. Since the production of bioethanol and its by-products is growing worldwide, including in Brazil, mycotoxicological monitoring of maize DDGS is crucial to identify the effects of mycotoxins occurrence in animal feed formulated with this ingredient.

Tracking Zearalenone and Type-B Trichothecene Mycotoxins in the Commercial Production of Beer and Brewers’ Spent Grains

The contamination of brewers’ spent grains (BSG) with mycotoxins may pose a significant threat to the health of domestic animals that consume BSG as feed. New information is needed regarding the presence of mycotoxins throughout commercial beer and BSG production. Samples (n = 106) were collected during production of a single batch of commercial beer, including malt (n = 54), mash (n = 6), wort (n = 6), fermentation, (n = 14), beer (n = 3), and BSG (n = 23) and analyzed for the presence of deoxynivalenol (DON), 3-acetyldeoxnivalenol (3ADON), 15-acetyldeoxynivalenol (15ADON), and zearalenone (ZEN) using gas chromatography–mass spectrometry (GC-MS). DON, 3ADON, 15ADON, and ZEN were observed at low levels (<0.05 mg kg−1) throughout the production of the beer. Despite most of the samples having low levels of mycotoxins, qualitative analyses showed trace levels of DON, 15ADON, and ZEN in barley malt, solid-fractions of mash, and BSG. Overall, results from this study suggest that low-levels of mycotoxins are retained in BSG, which may consumed by livestock. Results may help brewers and animal producers to consider storage and feeding procedures for BSG.

Microbiological analysis of water and fish of fish farms in the protection area of the western lowlands of Maranhão, Brazil

This work aimed to analyse the water quality and identify the fungal diversity present in the skin and gills of tambatinga fish from the environmental protection area of the western lowlands of Maranhão, Brazil. The physicochemical parameters of water were analysed and microbiological tests were carried out to identify the most probable number of total coliforms and Escherichia coli. Forty tambatinga specimens were collected. In addition, skin and gill fragments were removed for fungi isolation in culture medium (BDA). Physicochemical and microbiological parameters of water, fish biometry and fungal diversity of fish in the dry and rainy periods were analysed. Results showed significant differences in the total length (TC) and weight (P) of fish in the dry and rainy periods. Microbiological data showed that the water of the four fish farms analysed in both periods are within the limits established by the National Environment Council (CONAMA) of Brazil and within limits established in complementary literature. Regarding the physicochemical parameters of water, only conductivity (F (1, 14) = 8.611; p = 0.011) and temperature had considerable differences (F (1, 14) = 11.037; p = 0.005). There was prevalence of yeasts (33%) and fungi of the genus Fusarium sp. (26%), Penicillium sp. (21%) and Aspegillus sp. (20%). The analysis of fungal diversity showed associations for sex, Aspegillus sp. (X2 = 19,600; DF = 1; p = 0,001) and the dry season, Penicillium sp. (X2 = 8,120; DF = 1; p = 0,004) and Fusarium sp. (X2 = 21.583; GL = 1; p = 0.001). It is concluded, therefore, that the harvests are within the established by the legislation of the National Environment Council—CONAMA in Brazil and complementary literature. However, it is necessary to evaluate the presence of possible mycotoxins in cultivated fish, since the most prevalent fungal species are considered to be mycotoxigenic fungi and are widely disseminated in the environment.