Toxigenic Fungi And Fungal Mycotoxins In Food

Growth of prevalent filamentous fungi in foods may result in the production of mycotoxins, which can cause a variety of adverse effects in humans, including allergic reactions, immunosuppression, and cancer. Mycotoxins aflatoxins, ochratoxin A, fumonisins, tnchothecenes, and zearalenone are the most significant. Aflatoxins are potent carcinogens and, in conjunction with the hepatitis B virus, are responsible for tens of thousands of human fatalities annually, primarily in tropical non-industrialized nations.

Ochratoxin A is a likely carcinogen and may induce cancer of the urinary tract and kidney damage in northern and eastern Europeans. Fumonisins are believed to be the cause of oesophageal cancer in southern Africa, sections of China, and other locations. Trichothecenes are extremely immunosuppressive, whereas zearalenone has oestrogenic effects on both animals and humans.

Current records and statistics do not reflect the significant role played by mycotoxins in deaths caused by foodborne microorganisms. Mycotoxins, which are produced by fungi commonly found in foods and animal supplies, have only become apparent in the past 30 years. Throughout history, these toxins have caused significant epidemics among humans and animals.

Ergotism, which killed hundreds of thousands of people in Europe in the last millennium1 ; alimentary toxic aleukia (ATA), which killed at least 100,000 Russians between 1942 and 19482 ; stachybotryotoxicosis, which killed tens of thousands of horses in the Soviet Union in the 1930 ; and aflatoxicosis, which killed 100,000 young turkeys in the United Kingdom in 1960 and has caused death and disease in other animals, arachnid

What are toxigenic fungi?

• Toxigenic fungi are fungi with the capacity to produce mycotoxins, which are toxic compounds. These mycotoxins pose a threat to human and animal health if ingested, inhaled, or absorbed through the epidermis. Mycotoxins are secondary metabolites generated as a defense mechanism or to compete with other microorganisms by certain species of fungi.
• Toxic fungi are capable of contaminating agricultural commodities, stored foods, and indoor environments. They can thrive on a variety of organic substances, including cereals, nuts, fruits, vegetables, and even building materials. Toxigenic fungi from the genera Aspergillus, Penicillium, Fusarium, and Alternaria are widespread.
• Mycotoxins produced by toxigenic fungi can cause a variety of adverse health effects, depending on the specific mycotoxin, the level and duration of exposure, and the type of mycotoxin. Common symptoms of mycotoxin exposure include respiratory issues, allergic reactions, skin irritation, nausea, vomiting, diarrhea, and in extreme cases organ damage or mortality.
• Preventing and controlling the growth of toxigenic fungi requires the proper storage and handling of food and agricultural products, the maintenance of good indoor air quality and ventilation, and the prompt resolution of any water leakage or excessive moisture problems that could promote fungal growth. In addition to regular building inspections and maintenance, appropriate sanitation practices can help reduce the risk of toxigenic fungal contamination.

What are Fungal Mycotoxins?

Fungal mycotoxins are toxic compounds produced by certain species of filamentous fungi under conditions of high temperature and humidity. These fungi grow on food during long-term storage and transportation, leading to contamination with mycotoxins. Here are some key points about fungal mycotoxins:

1. Mycotoxin production: Mycotoxins are secondary metabolites produced by fungi at the end of their exponential growth phase. Fungi use specialized pathways and specific genes to produce these toxic compounds.
2. Toxicity to humans and animals: Mycotoxins are low molecular weight compounds that are harmful to humans and animals. They can cause food poisoning illnesses, teratogenic effects (birth defects), cancers, and damage to the kidneys and liver.
3. Fungal species and mycotoxin production: Various fungal species are associated with mycotoxin production. Aspergillus and Penicillium species, including A. flavus, A. ochraceus, A. parasiticus, A. niger, A. nomius, P. verrucosum, P. nordicum, P. dipodomyis, and P. chrysogenum, grow on crops during storage and produce toxins. Fusarium and Claviceps species, such as F. cerealis, F. oxysporum, F. proliferatum, F. verticilloides, F. sporotrichioides, F. moniliforme, F. graminearum, and Claviceps purpurea, infect crops before harvest. Other mycotoxin-producing fungi include Alternaria, Trichothecium, Cladosporium, Byssochlamys, and Sclerotinia.
4. Influencing factors: The growth and mycotoxin production of these fungi are influenced by various extrinsic conditions, including moisture, temperature, pH, and water activity. High moisture levels and warm temperatures favor fungal growth and mycotoxin production.
5. Susceptible food products: Most cereal and grain products, such as corn, wheat, rice, sorghum, maize, soybean, as well as certain spices (chili peppers, black peppers, coriander), and various nuts like pistachio, almond, walnut, peanut, and Brazil nuts, are susceptible to fungal infection and mycotoxin contamination.
6. Mold presence vs. mycotoxin presence: It’s important to note that the presence of mold in food products does not necessarily indicate the presence of mycotoxins. Conversely, mycotoxins may be present even if mold is not visibly present since the toxins can persist long after the mold has disappeared.

Fungal Mycotoxins Epidemiology (Mycotoxicoses)

The epidemiology of fungal mycotoxicoses, referring to the outbreaks and diseases caused by fungal mycotoxins, can be understood by considering the following points:

1. Outbreaks in recent years: Several outbreaks of mycotoxicoses have been reported in recent years, affecting both humans and domestic animals. These outbreaks are often linked to the consumption of long-term stored molded food and feed that contains toxigenic mycotoxins.
2. Historical outbreaks: The occurrence of mycotoxicoses can be traced back to the Middle Ages, where thousands of people were affected by a disease known as St. Anthony’s fire. This disease was later attributed to the fungus Claviceps purpurea, which parasitizes rye and other plants.
3. Mycotoxicoses during World War II: During World War II, a human disease outbreak occurred in Russia due to the consumption of long-term stored moldy grain. This resulted in various symptoms, including dermal necroses, hemorrhages, leucopenia, and bone marrow degradation.
4. Mycotoxicoses in animals: Animals are also susceptible to mycotoxicoses. In Japan, serious liver damage was observed in animals that consumed feed containing mold-contaminated yellowed rice. Various Penicillium species, such as P. citreo-viride, P. citrinum, P. islandicum, and P. rugulosum, were isolated from the yellow rice samples.
5. Aflatoxin outbreak in England: The presence of mycotoxins gained significant attention during a large outbreak that occurred in turkey poults in England. The contaminated feed component contained peanut meal, from which the fungus Aspergillus flavus was isolated. The toxic compound produced by this fungus was named aflatoxin.

These historical and more recent outbreaks highlight the impact of fungal mycotoxicoses on human and animal health. Improved storage and handling practices, as well as proper monitoring and control measures in the agricultural and food industries, are crucial for preventing and mitigating the risks associated with fungal mycotoxins.

Clinical manifestations of Mycotoxins

Mycotoxins can give rise to various clinical manifestations, encompassing acute, chronic, mutagenic, and teratogenic effects. Here are some key points about the clinical manifestations of mycotoxins:

1. Acute toxicity: Acute poisoning is the most commonly observed effect of mycotoxin exposure. It can cause significant damage to the liver and kidneys, and in extreme cases, it can result in patient death.
2. Food poisoning illnesses: Mycotoxicoses often present with symptoms resembling food poisoning. These can include vomiting, diarrhea, abdominal pain, fever, chronic fatigue, skin rashes, insomnia, depression, and anxiety. These symptoms may arise shortly after the ingestion of contaminated food.
3. Chronic toxicity: Prolonged exposure to mycotoxins can lead to chronic toxicity, resulting in various severe effects on the body. These effects can include carcinogenicity (the potential to cause cancer), skin necrosis (tissue death), leucopenia (reduced white blood cell count), autoimmune diseases, neurological issues, chronic organ damage, and static shocks (unusual sensations or discharges of static electricity).

It is important to note that the specific clinical manifestations of mycotoxin exposure can vary depending on the type of mycotoxin involved, the level and duration of exposure, individual susceptibility, and other factors.

Prompt recognition and appropriate management of mycotoxin-related illnesses are crucial for minimizing the impact on affected individuals. Seeking medical attention if symptoms occur after suspected mycotoxin exposure is essential for accurate diagnosis and appropriate treatment.

Toxigenic fungi

Toxigenic fungi belong to various fungal genera, with some of the most notable ones being:

• Aspergillus: This genus contains the species Aspergillus flavus, Aspergillus parasiticus, and Aspergillus ochraceus. Aspergillus flavus and Aspergillus parasiticus are known to produce aflatoxins, which are powerful carcinogens and can contaminate peanuts, maize and tree nuts. Aspergillus ochraceus generates the mycotoxin ochratoxin A, which is linked to kidney damage.
• Fusarium: Fusarium species, including Fusarium graminearum, Fusarium culmorum, and Fusarium verticillioides (previously known as Fusarium moniliforme), produce numerous mycotoxins. Deoxynivalenol (DON), zearalenone (ZEN), and fumonisins are examples. Mycotoxins can contaminate cereals such as wheat, maize, and barley, and have been linked to a variety of adverse health effects in humans and animals.
• Penicillium: Species of Penicillium, such as Penicillium verrucosum and Penicillium expansum, can generate mycotoxins like ochratoxin A, which can contaminate cereals, coffee, and other food products. Some Penicillium species are also responsible for the production of patulin, a mycotoxin found in rotten fruits, especially apples.
• Alternaria: Alternaria is known to produce mycotoxins, including alternariol and alternariol monomethyl ether. These mycotoxins can contaminate cereals, fruits, and vegetables, among other crops.
• Claviceps: The fungus Claviceps purpurea generates the mycotoxin ergotamine, which is linked to the human and animal disease ergotism. Fungus-infected cereals, particularly rye, are a common source of ergotamine.
• Stachybotrys: Stachybotrys chartarum, also known as black mould, is capable of producing mycotoxins known as trichothecenes. Satratoxin, a well-known trichothecene produced by Stachybotrys, has been linked to respiratory symptoms and other adverse human health effects.
• Trichothecium: Trichothecium species are capable of producing trichothecene mycotoxins, such as trichothecin and verrucarin. These mycotoxins have been discovered in a variety of foods, including grains and fruits.
• Chaetomium: Chaetomium species, such as Chaetomium globosum, can produce chaetoglobosins, which are mycotoxins. These mycotoxins have been linked to human cell toxicity and may contribute to domestic air quality problems.
• Fusarium culmorum and Fusarium poae: Fusarium culmorum and Fusarium poae are known to produce the mycotoxin beauvericin. Beauvericin has been associated with numerous toxicological effects and has been identified in cereals, particularly wheat.
• Acremonium: Acremonium species are capable of producing a variety of mycotoxins, including trichothecenes, fumonisins, and moniliformin. Grains, legumes, and spices have been found to contain these mycotoxins.
• Myrothecium: Myrothecium species are capable of producing mycotoxins such as roridin and verrucarin. These mycotoxins have been identified in contaminated grains and are toxic to humans and animals.
• Neosartorya and Aspergillus niger: Neosartorya and Aspergillus niger are known to produce ochratoxin A, which can contaminate a wide range of food products, including cereals, coffee, and preserved fruits.

Transmission of Fungal Mycotoxins

Humans and animals can be exposed to mycotoxins through a variety of routes. Transmission depends on the type of mycotoxin, the contaminated food or feed source, and the routes of exposure. Here are some typical transmission methods:

• Ingestion: The most prevalent route of mycotoxin exposure is through the consumption of contaminated food or forage. Mycotoxins are capable of contaminating a vast array of agricultural products, including cereals, nuts, fruits, and animal feed. Mycotoxins can be absorbed by the digestive system and enter the bloodstream when these contaminated products are ingested, potentially causing adverse health effects.
• Inhalation: Certain mycotoxins are capable of becoming airborne and can be found in dust particles or spores. Inhalation of contaminated particles or spores can result in mycotoxin exposure through the respiratory system. Occupational settings, such as agricultural employees handling moldy crops or workers in mold-contaminated environments, are particularly susceptible to this mode of transmission.
• Dermal Absorption: Certain mycotoxins, including specific varieties of aflatoxins, can be absorbed through the skin. This is a less common mode of transmission than ingestion or inhalation, but it can occur in occupational contexts involving direct contact with mycotoxin-contaminated materials or surfaces.
• Maternal-Fetal Transfer: In pregnant animals or humans, mycotoxins ingested by the mother can cross the placenta and be conveyed to the fetus, resulting in prenatal exposure. This may have serious health consequences for the maturing fetus.
• Breast Milk: Mycotoxins can be excreted into breast milk by lactating humans and animals. If the mother has been exposed to mycotoxins through contaminated food or environmental sources, the infant may be exposed through breastfeeding.

Mycotoxins Produced by Toxigenic fungi

1. Aflatoxins: Aflatoxins are predominantly produced by the fungi Aspergillus flavus and Aspergillus parasiticus. Among them are aflatoxin B1, B2, G1, G2, M1 and M2. Aflatoxins are potent carcinogens and have been linked to liver cancer, immune suppression, and other health problems. Typically, they contaminate peanuts, sorghum, cottonseed, and tree nuts.
2. Ochratoxin: Several species of fungi, including Aspergillus ochraceus and Penicillium verrucosum, produce ochratoxins, specifically ochratoxin A, B, and C. Ochratoxin compounds are nephrotoxic and can cause kidney injury. They can contaminate cereals, coffee, wine, and preserved fruits, among other food products.
3. Trichothecenes: Trichothecenes are a type of mycotoxin generated by a variety of fungi, including Fusarium, Myrothecium, and Stachybotrys. Trichothecenes are represented by deoxynivalenol (DON), T-2 toxin, HT-2 toxin, and nivalenol. Trichothecenes can have a variety of toxic effects, such as immunosuppression, gastrointestinal problems, dermatitis, and interference with protein synthesis.
4. Zearalenone: Fusarium species, such as Fusarium graminearum and Fusarium culmorum, produce zearalenone (ZEN). It has estrogenic properties and can interfere with animal reproduction. ZEN is frequently found in cereals, especially maize.
5. Fumonisins: Several Fusarium species, including Fusarium verticillioides and Fusarium proliferatum, produce fumonisins. Fumonisins, such as fumonisin B1, B2, and B3, can cause a variety of health problems, such as liver and kidney injury, neurotoxicity, and cancer. They can contaminate maize and products made from maize.
6. Patulin: Patulin is produced by Penicillium expansum and Penicillium griseofulvum primarily. It is frequently found in mouldy fruits, especially apples, and can cause gastrointestinal irritation and liver injury.
7. Ergot Alkaloids: Claviceps purpurea, a fungus that infects cereal grains, especially rye, produces ergot alkaloids. Symptoms of ergotism include hallucinations, vasoconstriction, and gangrene. Ergot alkaloids, including ergotamine and ergocristine, can induce ergotism.
8. Citrinin: Several fungal species, including Penicillium, Aspergillus, and Monascus, produce citrinin. It is capable of contaminating a vast array of food products, including cereals, rice, and fermented goods. Citrinin has been linked to nephrotoxicity and is capable of causing kidney injury.
9. Patulin: In addition to Penicillium species, certain Aspergillus and Byssochlamys species are also capable of producing patulin. Patulin is commonly found in mouldy fruits, particularly apples and products containing apples. It may possess genotoxic and immunotoxic properties in addition to toxic effects on the gastrointestinal system.
10. Sterigmatocystin: Aspergillus species, including Aspergillus versicolor and Aspergillus nidulans, produce sterigmatocystin. It has a similar chemical structure to aflatoxins and is found in cereals, nuts, and some seasonings. It is believed that sterigmatocystin has carcinogenic properties, notably in relation to liver cancer.
11. Penitrem A: Penicillium species, including Penicillium crustosum and Penicillium commune, produce Penitrem A. It is frequently found in mouldy foods, especially cereals and nuts. Penitrem A is a neurotoxin that can cause animals to exhibit tremors, convulsions, and other neurological symptoms.
12. Zearalenols: Fusarium species also produce zearalenols, which are derivatives of zearalenone. -zearalenol and -zearalenol are among them. Comparable to zearalenone in their estrogenic properties, these mycotoxins can affect the reproductive health of animals.
13. Aurovertins: Several Aspergillus species, including Aspergillus ochraceus and Aspergillus versicolor, produce aurovertins. They have been detected in cereals, fruits, and dairy products. Aurovertins are capable of inhibiting mitochondrial function and exerting cytotoxic effects.
14. Penicillic Acid: Penicillium species, including Penicillium camemberti and Penicillium roqueforti, produce penicillic acid. It is frequently detected in contaminated cheese and other dairy products. Nephrotoxicity and genotoxicity have both been linked to penicillic acid.
15. Rubratoxins: Penicillium rubrum and Penicillium purpurogenum produce rubratoxin. Cereals and grains have been found to contain these mycotoxins.

How Fungal Mycotoxins Cause Infection?

Mycotoxins are primarily responsible for toxic effects rather than infection. They are secondary metabolites produced by specific moulds (fungi) and are toxic to humans and animals when consumed or otherwise exposed. Mycotoxins are not infectious agents; rather, they are chemical compounds with potentially detrimental effects on the body.

Mycotoxins can be absorbed through the digestive tract and enter the circulation after consumption of mycotoxin-contaminated food or feed. They can then interact with various organs and tissues, causing toxic effects. Depending on the variety of mycotoxin and the target organs or systems affected, the precise mechanisms by which mycotoxins cause harm can vary. Here are some examples of how mycotoxins can cause toxicity:

• Damage to DNA and Proteins: Certain mycotoxins, including aflatoxins and ochratoxins, can bind to DNA or proteins within cells, causing structural damage or interfering with normal cellular processes. This may result in mutations, cell demise, or cellular function disruption.
• Inhibition of Enzymes: Certain mycotoxins are capable of inhibiting vital enzymes involved in cellular metabolism or other physiological processes. This disruption of enzyme activity can disrupt normal cellular functions and contribute to toxicity.
• Oxidative Stress: Mycotoxins can elicit oxidative stress in cells, resulting in an imbalance between the production of reactive oxygen species (ROS) and the antioxidant defence system of the body. This oxidative stress can cause damage to cellular components, such as lipids, proteins, and DNA, and contribute to inflammation and tissue damage.
• Immunosuppression: Certain mycotoxins, including deoxynivalenol (DON) and fumonisins, can suppress the immune system, reducing its ability to combat infections and increasing susceptibility to other diseases. This immunosuppressive effect may increase susceptibility to opportunistic infections.
• Carcinogenesis: Certain mycotoxins, including aflatoxins and some types of ochratoxins, have been classified as carcinogens and have been shown to increase the risk of developing cancer. These mycotoxins can induce DNA mutations, interfere with cellular signalling pathways, and promote the development and progression of tumours.

Notably, the severity of mycotoxin toxicity can vary depending on the type of mycotoxin, the dose and duration of exposure, individual susceptibility, and overall health status. To protect public health and reduce the risk of mycotoxin-related toxicity, regulatory agencies impose maximum mycotoxin levels in food and animal feed.

A summary of the mechanisms underlying the exacerbating effects of mycotoxin exposure when the immune system is dysregulated.

(a) Mycotoxins in the pathogenesis of multiple sclerosis: exposure to GTX modifies the blood-brain barrier. Mycotoxins damage astrocytes, oligodendrocytes, and microglia in neural tissue. Loss of oligodendrocytes increases demyelination, while targeted astrocytes release proinflammatory cytokines that contribute to the neuroinflammatory milieu. Induction of proinflammatory gene expression in the CNS is a further direct consequence of mycotoxins. It is hypothesized that indirect pathways involving proinflammatory cytokines such as IL-1β interact with microglia via an increased kynurenine/tryptophan ratio, which promotes the secretion of neurotoxic metabolites.

(b) Exposure to mycotoxins worsens respiratory epithelium barrier impairment in asthmatic conditions. Mycotoxin uptake by dendritic cells results in a decrease in IL-12 production, an increase in ROS production, and an overactivation of the inflammasome. Reduction in IL-12 accentuates the Th1/Th2 imbalance contributing to increased airway inflammation in a mouse model of asthma.

(c) Potential modes of action between mycotoxin exposure and HIV replication: mycotoxin exposure modifies immune response by inducing reactive oxygen species (ROS). ROS inhibits oxidative defense machinery by retaining Nrf2 and inducing proinflammatory response by inducing NF-B. Mycotoxin-related oxidative stress and proinflammatory signals could both potentially contribute to an increase in HIV prevalence and disease progression.

Fungal Mycotoxins Detection Methods

• Immunoassays: Due to their simplicity, sensitivity, and swift results, immunoassays are widely used for mycotoxin detection. Among the most prevalent immunoassay methods are enzyme-linked immunosorbent assays (ELISAs) and lateral flow devices. These techniques rely on antibodies that bond to specific mycotoxins and generate a detectable signal.
• Chromatographic techniques: Chromatography is a highly sensitive and specific technique for mycotoxin analysis. Common chromatographic techniques include gas chromatography (GC) and liquid chromatography (LC). High-performance liquid chromatography (HPLC) and ultra-high-performance liquid chromatography (UHPLC) are subcategories of liquid chromatography (LC). Chromatographic techniques are frequently coupled with a variety of detectors, such as mass spectrometry (MS) or ultraviolet-visible (UV-Vis) spectrophotometry, to improve their detection capabilities.
• Mass spectrometry: Mass spectrometry is an extremely sensitive and selective technique for identifying and quantifying mycotoxin. To improve the analysis, it can be combined with various separation techniques such as gas chromatography (GC-MS) or liquid chromatography (LC-MS). Mass spectrometry provides precise molecular weight information and can simultaneously detect multiple mycotoxins.
• Nucleic acid-based methods: To detect specific fungal species or genes responsible for mycotoxin production, polymerase chain reaction (PCR) and other nucleic acid amplification techniques may be utilized. These techniques can provide early detection of fungal contamination and aid in predicting the likelihood of mycotoxin formation.
• Biosensors: Biosensors are analytical devices that combine biological components like enzymes, antibodies, or microorganisms with physicochemical transducers. They provide rapid, sensitive, and portable mycotoxin detection. Depending on the transducer employed, biosensors can be founded on various principles, such as electrochemical, optical, or piezoelectric.
• Near-infrared (NIR) spectroscopy: NIR spectroscopy is a nondestructive and rapid technique that can be used for mycotoxin analysis. It relies on the near-infrared absorption of light by chemical bonding in the sample. Statistical models can be devised to predict the presence and concentration of mycotoxins by analyzing spectral data.

Prevention and control measures of mycotoxin in food

• Good Agricultural Practices (GAP): It is essential to implement appropriate agricultural practices to reduce fungal growth and mycotoxin contamination in crops. This includes the use of certified seeds, crop rotation, monitoring irrigation practices, and effective insect management.
• Harvesting and Post-Harvest Practices: For the prevention of mycotoxin formation, it is essential to harvest crops at the appropriate time and to ensure adequate drying and storage conditions. To prevent the development of fungi, harvested crops should be dried to an appropriate level of moisture content. Utilize proper storage facilities with adequate ventilation, temperature regulation, and humidity prevention.
• Integrated Pest Management (IPM): IPM practices can help reduce the use of chemical pesticides and fungal contamination by minimizing the application of chemical pesticides. IPM combines techniques such as crop rotation, biological control agents, and cultural practices to effectively manage pests.
• Good Manufacturing Practices (GMP): To reduce the risk of mycotoxin contamination, food processing facilities must adhere to GMP guidelines. This includes maintaining sanitary conditions, implementing appropriate cleaning and sanitation procedures, and assuring the proper storage and handling of raw materials.
• Hazard Analysis and Critical Control Points (HACCP): Hazard Analysis and Critical Control Points (HACCP) is a systematic approach used to identify and control potential food production hazards. Identifies critical control points where mycotoxin contamination can occur and implements control measures to prevent or eliminate contamination at those points.
• Quality Control and Testing: Regular testing and monitoring for mycotoxin contamination of basic materials, intermediate products, and finished food products is required. This aids in the early detection and avoidance of contaminated products reaching consumers. For mycotoxin analysis, techniques such as immunoassays, chromatography, and mass spectrometry can be utilized.
• Education and Training: Education and Training It is essential to educate and train farmers, food handlers, and food processors about the risks associated with mycotoxins and the correct preventative measures. This contributes to raising awareness and assuring the implementation of appropriate practices throughout the entire food production chain.
• Regulatory Standards: Governments and regulatory agencies establish maximum levels of mycotoxin in food and animal feed. Compliance with these regulations contributes to the control of mycotoxin contamination and the protection of consumers.

FAQ

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