Monday, September 28, 2020

Common Foodborne Pathogens - IV

Aflatoxins Risk Profile

Mycotoxins are produced by mold, and there is a range of different compounds originating from different types of mold among the Deuteromycetes. The Mycotoxins are secondary metabolites, toxic in low concentrations in vertebrates.
 

Aflatoxins are a group of mycotoxins produced by Aspergillus flavus and Aspergillus parasiticus and can cause serious complications in animals and humans. The four major aflatoxins are AFB1, AFB2, AFG1, and AFG2. These may grow in a broad range of agricultural commodities including plant leaves, dried fruit, corns, nuts, and infect stored cereal grains where they produce the aflatoxins. When such toxins are formed they do not go away. The aflatoxin was first recognized in damaged peanuts contaminated with Aspergillus flavus. The aflatoxins are easily identifiable with their blue or green fluorescence under UV light and relative chromatographic mobility after thin-layer chromatographic separation. They are heat-stable and thus stay in the food along the food chain, unaffected by heat treatments such as pasteurization. On the other hand, aflatoxin AFB1, which is present in contaminated feed or food can be converted to M1 (AFM1). If a cow eats a feed contaminated with aflatoxin B1, the activity of the cow changes the aflatoxin B1 to aflatoxin M1, which ends up in the milk (Johnsson, 2006). Hence, AFM1 in milk is not as hazardous as its parent compound, but the limit is 0.5 parts per billion, largely because milk tends to constitute a large part of the diet of infants and children.

 

The toxicity of aflatoxin is mainly due to its carcinogenicity and the most potent known natural carcinogen is AFB1. This is because aflatoxins are genotoxic, meaning it affects the genetic material. Genotoxins have a direct dose-response relationship, so they do not have a threshold dose to exceed before they have an effect. Thus, there is no tolerable daily intake (TDI) for aflatoxins, which are to be kept at a level as low as possible. 

 

Sources

Aflatoxins are frequently found in many serials including corn/maize, sorghum, rice, cottonseed, peanuts, tree nuts, copra, cocoa beans, nutmeg, dried fruit such as figs, rhizomes such as ginger and plant leaves, as well as AFM1 may be found in milk and dairy products. Nonetheless, Ghana, Kenya, Nigeria, Sudan, Thailand, and other developing countries have reported several cases of Aflatoxin M1 found in human breast milk due to the mother’s chronic exposure to dietary aflatoxins.

Disease

Chronic exposure to aflatoxin affects many organs and it critically affects the liver as the body’s detoxification functions are usually taken place in the liver because aflatoxins are hepatotoxic in humans and animals. Long-term aflatoxin exposures from the food products may cause aflatoxicosis, which can range from acute to chronic, and sickness can range from mild to severe. The severe liver damage can lead to cirrhosis, which may lead to the development of liver cancer, but it is usually not possible to prove the damage is caused by aflatoxins, hence test tumor tissue for biomarkers or characteristic genetic damage are the basic possible models.


Mortality:

There are several cases of documented aflatoxin poisoning around the world including India (1974), Kenya (1982, 2004 and 2005), and Malaysia (1988) resulting in altogether 747 hospitalizations and 258 fatalities. Thus, by far it presences a great danger to humans and animals in the context of food safety and it is lethal in both acute and chronic exposures, given the right dose depend on the exposing person’s or animal’s general health status.

 

Toxic dose:

The exact toxic dose for a human is yet unknown, but there are documented records of toxic levels of aflatoxin ingestions from in the following countries illustrate different mortality rates from outbreaks:

o   In 1974, there were 397 sufferers and 108 fatalities in northwest India, where aflatoxin levels of 0.25 to 15 mg/kg were found in corn.

o   In 1982, in Kenya reported 20 hospitalizations with a 60% mortality rate, with aflatoxin intake of 38 µg/kg of body weight.

o   In 1988, 13 Chinese children died after eating Chinese noodles in Malaysia, due to acute hepatic encephalopathy, where postmortem samples from the patients confirmed the presence of aflatoxins.

o   One of the largest aflatoxicosis outbreaks on record occurred during 2004 and 2005 in rural Kenya, exposing 317 people with 125 fatalities, where the root cause was aflatoxin-contaminated homegrown maize with an average toxic dose of 354 ng/g.  

o   A laboratory worker who intentionally ingested AFB1 at 12 µg/kg body weight for two days has developed a rash, nausea, and headache, but recovered without ill effect. However, a 14-year follow-up study of the worker confirmed that physical examination and blood chemistry, including tests for liver function, were normal.

o   The effects of aflatoxins on the health of the various animals depend on their species, level, and duration of exposure, including nutritional status. Thus, the median lethal dose (i.e., LD50) values show wide variation, ranging from 0.3 mg/kg body weight in rabbits to 18 mg/kg body weight in rats.

o   Aflatoxins do not affect equally in every animal, but moderately to highly toxic and carcinogenic in almost every animal species tested, and their main factor for tolerance relates to the nature of the digestive system. Other tolerance factors include breed variety, nutrition, sex, age, environmental stress, and the presence of other disease agents, and ruminants are more tolerant. But other animals such as swine, chickens, ducks, pets, and wild birds are more sensitive.

 

Onset:

Not applicable.


Complications:

Acute exposure to high doses of aflatoxin can result in aflatoxicosis which leads to the damage of the liver where aflatoxin inhibits the normal functions of the liver, including carbohydrate and lipid metabolism and protein synthesis. Cancer, impaired protein formation, impaired blood coagulation, toxic hepatitis, and probable immunosuppression can be resulted from chronic exposure due to substantial doses. There can be reduced weight gain and reduced feed-conversion efficiency in animals apart from the complications. The International Agency for Research on Cancer has classified AFB1 as a group 1 carcinogen and AFM1 as a group 2b carcinogen (carcinogenic to laboratory animals and possibly carcinogenic to humans, respectively), because AFB1 is the most potent known natural carcinogen and is the most abundant of the aflatoxins and they are probably immunosuppressive in humans. Combined exposure to aflatoxin and hepatitis B increases the risk for the development of human hepatocellular carcinoma (HCC) and the diagnosis of chronic aflatoxicosis is difficult without sophisticated laboratory equipment. Aflatoxins are primarily affecting the cellular immune processes and also affect to decrease in antibody formation, embryonic exposure, and reducing immune responses in offspring, in most of the laboratory animal species studied.


Symptoms:

General symptoms include edema of the lower extremities, abdominal pain, and vomiting in addition to the disruption and inhibition of carbohydrate and lipid metabolism and protein synthesis associated with aflatoxicosis in humans can lead to hemorrhaging, jaundice, premature cell death, tissue necrosis in the liver and, possibly other organs.


Duration:

The duration of symptoms are varied, but no appropriate scientific data is available.

 

Route of entry:

Oral.

 

Pathway:

AFB1 can interact with DNA, leading to structural damages which if not repaired, a mutation can occur that may initiate the cascade of events required to produce cancer. Once activated by cytochrome P450 monooxygenases, AFB1 is metabolized to a highly reactive metabolite, AFB1-exo-8,9-epoxide. Then metabolize binds with the guanine moiety of DNA at the N7 position, forming trans-8,9-dihydro-8-(N7-guanyl)-9­ hydoxyAFB1 adducts, which can rearrange and form a stable adduct, and studies have shown that it is associated with tumor cells.


Frequency

According to the Worldwide Regulations for Mycotoxins 2003, more than 76 countries have legislated limits on aflatoxins, ranging from 0 to 35 ng/g. More acute and chronic exposures were reported in developing countries where no regulatory limits, poor agricultural practices in food handling and storage, malnutrition, and disease. However, aflatoxin contamination in foods at levels that can cause acute aflatoxicosis in humans has rarely occurred in developed countries.
 

Diagnosis

The aflatoxicosis in humans can be diagnosed by Jaundice and its characteristic yellowing of tissues due to the liver damage, as well as gall bladder may become swollen, and immunosuppression may provide an opportunity for secondary infections. There can be a decrease in vitamin K functions and high levels of AFB1-albumin adducts may be present in plasma. Aflatoxin exposure can be identified through biomarkers that detect the presence of metabolites in blood, milk, and urine, and excreted DNA adducts and blood-protein adducts. Further, AFB1-albumin adducts can be measured in blood and converted AFM1 and AFB1-DNA adduct or AFB1-guanine adduct can be subsequently detected in the urine of people consuming sufficient amounts of AFB1.

 

Target Populations
Susceptibility to aflatoxin of humans can vary with sex, age, health, nutrition, environmental stress, and level and duration of exposure, where aflatoxin-induced chronic and the acute syndrome is common in children and adults in some developing countries.

 

Food Analysis

The most difficult step in mycotoxin determination is the sampling due to contaminant variability and there are developed procedures for sampling, sample preparation, extraction, purification, isolation, separation, and quantitation of aflatoxins in foods. The use of proper sampling equipment and techniques can reduce the effects of sample selection while increasing sample size can reduce the effects of the distribution of contaminated particles within a lot.

 

Analytical methods used to identify aflatoxins are usually quantitative or semi-quantitative assays and rapid screening tests, where Thin-layer chromatography (TLC) is among the most widely-used analytical methods. Sample cleanup is a time-consuming step and usually consists of extraction with solvent, liquid-liquid partition, and/or chromatographic separation and determination. The antibody development is another technique that has led to the development of enzyme-linked immunosorbent assays (ELISAs) which are mainly used in screening methods. The high-performance liquid chromatography (LC) with fluorescence detection and hyphenated methods, such as LC/mass spectrometry (MS) or LC/MS-MS, are also used in quantitation and confirmation of identities. The modern analytical technologies evolving for aflatoxin detection includes solid-phase micro-extraction, surface-plasmon resonance, fiber-optic sensors, electrochemical immunosensors, fluorescence-based immunoassays, and the use of molecularly imprinted polymers.


Reference:

FDA Bad Bug Book, Foodborne Pathogenic Microorganisms and Natural Toxins. Second Edition. 2013

Preventive Controls for Human Foods. 2016

www.cdc.gov

 

 

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