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Aflatoxin B1
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Aflatoxin B1

Aflatoxin B1 (AFB1) is the most toxic and carcinogenic of the aflatoxins, a group of mycotoxins produced by molds like Aspergillus flavus and Aspergillus parasiticus. These molds thrive in warm, humid environments and commonly contaminate crops such as peanuts, corn, cottonseed, tree nuts, and spices. 

AFB1 is highly potent, capable of causing acute toxicity with symptoms like liver failure and hemorrhaging, and chronic exposure significantly raises the risk of liver cancer by inducing DNA mutations. 

While one may test for AFB1 presence directly, a primary biomarker for assessing AFB1 exposure is Aflatoxin M1 (AFM1), a hydroxylated metabolite found in urine.  Other biomarkers include Aflatoxin-N7-guanine (AF-N7-Gua) adducts, which indicate DNA damage, and minor metabolites like Aflatoxin Q1 (AFQ1) and Aflatoxin P1 (AFP1), which reflect the body's detoxification processes. 

These biomarkers are crucial for evaluating both immediate and long-term health risks associated with AFB1 exposure.

What are Aflatoxins?  [9.] 

Aflatoxins are a group of highly toxic and carcinogenic compounds produced as secondary metabolites by certain molds, primarily Aspergillus flavus and Aspergillus parasiticus.  [9., 20.]

They are one type of mycotoxin, which are a larger group of toxic chemical compounds produced by molds.  

These mycotoxins are formed when the molds colonize foods like peanuts, corn, cottonseed, tree nuts, and spices under favorable temperature and humidity conditions.   [9., 19.] 

The main aflatoxins are B1, B2, G1, and G2, with aflatoxin B1 (AFB1) being the most potent and carcinogenic.  [3.] 

Their production occurs when the mold's growth is stressed, such as during drought conditions or improper crop storage. 

Aflatoxins are among the most carcinogenic substances known - they can cause acute toxicity leading to liver failure, hemorrhaging, and even death in severe cases.   [9.]  Chronic exposure increases the risk of developing liver cancer, as aflatoxins can bind to DNA and cause mutations, particularly in the p53 tumor suppressor gene. [9., 11.]

They can also suppress the immune system, and can cause stunted growth in children.  [10., 15., 16.]

Worryingly, aflatoxins are highly stable and can persist in foods and animal feeds even after processing and cooking.  They bioaccumulate in the body over time, especially in the liver, due to their lipophilic nature and resistance to metabolic breakdown.  [1., 12., 14.] 

Aflatoxin B1 is metabolized to the reactive aflatoxin-8,9-epoxide, which binds to DNA and proteins, initiating carcinogenesis.  [16.] 

Animals fed contaminated feed can pass aflatoxin metabolites into meat, milk, and eggs, posing risks to humans consuming these products.

Anti-Aflatoxin Biomarkers

Anti-aflatoxin biomarkers are a group of compounds or metabolites that can be measured in biological samples to assess exposure to aflatoxins, which are toxic and carcinogenic metabolites produced by certain molds. 

These biomarkers provide a direct measure of internal exposure and can help evaluate the associated health risks.

A variety of biomarkers have been used to identify the presence of aflatoxins in humans and animals.  

Aflatoxin B1, known as one of the most potent and toxic aflatoxins, is one such biomarker.  

Aflatoxin B1 Biomarkers  [8.]

Aflatoxin B1 (AFB1) may be tested directly in urine; additionally, other biomarkers of AFB1 exposure can also be measured in urine samples:

Aflatoxin M1 (AFM1) 

AFM1 is a hydroxylated metabolite of AFB1, found in the milk of mammals consuming AFB1-contaminated feed.  This metabolite of AFB1 is commonly measured in urine to indicate recent AFB1 exposure.

Aflatoxin-N7-guanine (AF-N7-Gua) Adducts

These are products formed by the binding of AFB1 metabolites to guanine in DNA, and can be measured in urine to assess the biologically effective dose of AFB1 exposure leading to DNA damage.  It plays a key role in the initiation of liver cancer.  

AF-N7-Gua is an important biomarker as it indicates direct genotoxic impact of aflatoxin exposure.

Aflatoxin Q1 (AFQ1) and Aflatoxin P1 (AFP1)

AFQ1 is a hydroxylated metabolite of AFB1 formed during detoxification processes in the liver.  AFP1 is another detoxification product of AFB1, formed through similar pathways as AFQ1.

While less toxic than AFB1, the presence of AFQ1 indicates exposure and the body's detoxification response to aflatoxin.

AFP1 detection is part of assessing the metabolic processing of aflatoxins in the body.

Aflatoxin B1-Lysine Adduct  [4.]

The Aflatoxin B1-lysine adduct is formed when the reactive AFB1 epoxide binds to the ε-amino group of lysine residues in serum albumin or other proteins.  This adduct can be detected in blood or urine samples and serves as a reliable indicator of recent AFB1 exposure. 

Aflatoxin Q1 (AFQ1) and aflatoxin P1 (AFP1)  [9., 20.]

These are minor metabolites of AFB1 that can be measured in urine as additional biomarkers of recent exposure.

Aflatoxin B1 Laboratory Testing Methods

Test Information, Sample Collection and Preparation

Various biological samples can be employed to detect and quantify anti-aflatoxin biomarkers, particularly blood, urine and nasal secretions.

Blood samples are typically collected via venipuncture in a clinical setting, while urine and nasal secretion samples may be collected from the comfort of home.  

It is important to consult with the ordering provider prior to sample collection, as certain protocols may be recommended beforehand.  

Interpretation of Aflatoxin B1 Test Results

Optimal Levels of Aflatoxin B1 Test Results

Because of the high level of toxicity of aflatoxin B1, optimal test results indicate undetectable levels of aflatoxin B1.  

Clinical Significance of Elevated Aflatoxin B1

Elevated levels of aflatoxin B1 indicate recent or current exposure to aflatoxins.  However, because they are known to bioaccumulate, testing positive for the presence of aflatoxins may indicate a persistent historical exposure.  [5., 6., 18.]

Clinical Significance of Low Aflatoxin B1

Low or undetectable levels of aflatoxin B1 are considered ideal.  

Aflatoxin B1 Related Biomarkers

In addition to the direct measurement of aflatoxin B1 biomarkers, other biomarkers can provide complementary information about aflatoxin exposure and its potential health effects.

Oxidative Stress Markers  [17., 21.]

Aflatoxin exposure is known to induce oxidative stress, which can lead to cellular damage and contribute to the development of various diseases. 

Biomarkers such as malondialdehyde (MDA) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) can be measured to assess oxidative stress levels and the associated risk of aflatoxin-related toxicity.  [17.] 

Liver Function Markers  [7.] 

Since the liver is a primary target organ for aflatoxin toxicity, monitoring liver function biomarkers like alanine aminotransferase (ALT), aspartate aminotransferase (AST), and bilirubin can provide insights into the extent of hepatic injury and potential liver damage caused by aflatoxin exposure.  

Inflammatory Biomarkers  [17.] 

Aflatoxin exposure can trigger inflammatory responses, and biomarkers such as C-reactive protein (CRP), interleukins (e.g., IL-6, IL-8), and tumor necrosis factor-alpha (TNF-α) can be measured to evaluate the inflammatory status and associated health risks.

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See References

[1.] Bansal A, Sharma M, Pandey A, Shankar J. Aflatoxins: Occurrence, Biosynthesis Pathway, Management, and Impact on Health. Fungal Resources for Sustainable Economy. Published online 2023:565-594. doi:https://doi.org/10.1007/978-981-19-9103-5_21

[2.] Chen CH, Wang MH, Wang JH, et al. Aflatoxin Exposure and Hepatitis C Virus in Advanced Liver Disease in a Hepatitis C Virus–Endemic Area in Taiwan. American Journal of Tropical Medicine and Hygiene. 2007;77(4):747-752. doi:https://doi.org/10.4269/ajtmh.2007.77.747

[3.] Dai C, Tian E, Hao Z, Tang S, Wang Z, Sharma G, Jiang H, Shen J. Aflatoxin B1 Toxicity and Protective Effects of Curcumin: Molecular Mechanisms and Clinical Implications. Antioxidants (Basel). 2022 Oct 14;11(10):2031. doi: 10.3390/antiox11102031. PMID: 36290754; PMCID: PMC9598162.

[4.] de D, J. Solis-Mercado, M. Rodríguez-Aguilar, F. Díaz-Barriga, D. Guzmán Ortíz, R. Flores-Ramírez. Assessment of aflatoxin B1-lysine adduct in serum of infant population of the Huasteca Potosina, México – a pilot study. World mycotoxin journal. 2019;12(4):421-429. doi:https://doi.org/10.3920/wmj2019.2457

[5.] Ferri F, Brera C, De Santis B, Collini G, Crespi E, Debegnach F, Gargano A, Gattei D, Magnani I, Mancuso P, Mozzanica S, Teodori E, Djuric O, Giorgi Rossi P. Association between Urinary Levels of Aflatoxin and Consumption of Food Linked to Maize or Cow Milk or Dairy Products. Int J Environ Res Public Health. 2020 Apr 6;17(7):2510. doi: 10.3390/ijerph17072510. PMID: 32268619; PMCID: PMC7177871.

[6.] Gong YY, Watson S, Routledge MN. Aflatoxin Exposure and Associated Human Health Effects, a Review of Epidemiological Studies. Food Safety. 2016;4(1):14-27. doi:https://doi.org/10.14252/foodsafetyfscj.2015026

[7.] Güç, İ., Yalçin, E., Çavuşoğlu, K. et al. Toxicity mechanisms of aflatoxin M1 assisted with molecular docking and the toxicity-limiting role of trans-resveratrol. Sci Rep 12, 14471 (2022). https://doi.org/10.1038/s41598-022-18791-8

[8.] Hooper DG, Bolton VE, Guilford FT, Straus DC. Mycotoxin Detection in Human Samples from Patients Exposed to Environmental Molds. International Journal of Molecular Sciences. 2009;10(4):1465-1475. doi:https://doi.org/10.3390/ijms10041465

[9.] International Agency For Research On Cancer. Working Group On The Evaluation Of The Carcinogenic Risks To Humans. Some Traditional Herbal Medicines, Some Mycotoxins, Naphthalene and Styrene : This Publication Represents the Views and Expert Opinions of an IARC Working Group on the Evaluation of Carcinogenic Risks to Humans, Which Met in Lyon, 12-19 February 2002. Iarc; 2002.

[10.] Jiang Y, Jolly PE, Preko P, Wang JS, Ellis WO, Phillips TD, Williams JH. Aflatoxin-related immune dysfunction in health and in human immunodeficiency virus disease. Clin Dev Immunol. 2008;2008:790309. doi: 10.1155/2008/790309. PMID: 18695741; PMCID: PMC2496958.

[11.] Kew MC. Aflatoxins as a cause of hepatocellular carcinoma. J Gastrointestin Liver Dis. 2013 Sep;22(3):305-10. PMID: 24078988.

[12.] Kumar P, Mahato DK, Kamle M, Mohanta TK, Kang SG. Aflatoxins: A Global Concern for Food Safety, Human Health and Their Management. Front Microbiol. 2017 Jan 17;7:2170. doi: 10.3389/fmicb.2016.02170. PMID: 28144235; PMCID: PMC5240007.

[13.] Lalah, J.O., Omwoma, S., & Orony, D.A. (2019). Aflatoxin B1: Chemistry, Environmental and Diet Sources and Potential Exposure in Human in Kenya. Aflatoxin B1 Occurrence, Detection and Toxicological Effects.

[14.] MANCINI A, DREASSI E, BOTTA M, TARCHI F, FRANCARDI V. BIOACCUMULATION RISK ASSESSMENT OF AFLATOXIN B1, OCHRA-TOXIN AND FUMONISIN B1 IN TENEBRIO MOLITOR LARVAE. Redia. 2020;103:101-108. doi:https://doi.org/10.19263/redia-103.20.16

[15.] Mao Y, Dang M, Zhang J, et al. Peptide amphiphile inspired self-assembled, ordered gold nanocomposites for improved sensitivity of electrochemical immunosensor: Applications in determining the total aflatoxin amount in food stuffs. Talanta. 2022;247:123532-123532. doi:https://doi.org/10.1016/j.talanta.2022.123532

[16.] Mutocheluh M, Narkwa PW. Aflatoxin B1: An Immunomodulator and Cancer Agent. www.intechopen.com. Published November 30, 2022. https://www.intechopen.com/chapters/83715

[17.] Peng T, Li LQ, Peng MH, et al. Evaluation of oxidative stress in a group of adolescents exposed to a high level of aflatoxin B1 a multi-center and multi-biomarker study. Carcinogenesis. 2007;28(11):2347-2354. doi:https://doi.org/10.1093/carcin/bgm193

[18.] Tong Y, Tonui P, Orang'o O, Zhang J, Maina T, Muthoka K, Groopman J, Smith J, Madeen E, Ermel A, Loehrer P, Brown D. Association of Plasma Aflatoxin With Persistent Detection of Oncogenic Human Papillomaviruses in Cervical Samples From Kenyan Women Enrolled in a Longitudinal Study. Res Sq [Preprint]. 2023 Jan 27:rs.3.rs-2468599. doi: 10.21203/rs.3.rs-2468599/v1. Update in: BMC Infect Dis. 2023 Jun 6;23(1):377. PMID: 36747756; PMCID: PMC9901024.

[19.] Turner PC, Snyder JA. Development and Limitations of Exposure Biomarkers to Dietary Contaminants Mycotoxins. Toxins (Basel). 2021 Apr 28;13(5):314. doi: 10.3390/toxins13050314. PMID: 33924868; PMCID: PMC8147022.

[20.] Xu Y, Gong YY, Routledge MN. Aflatoxin exposure assessed by aflatoxin albumin adduct biomarker in populations from six African countries: REVIEW ARTICLE. World Mycotoxin J. 2018;11(3):411-419. doi: 10.3920/WMJ2017.2284. Epub 2018 Aug 1. PMID: 33552312; PMCID: PMC7797627.

[21.] Xue M, Fu M, Zhang M, et al. Aflatoxin B1 Induced Oxidative Stress and Gut Microbiota Disorder to Increase the Infection of Cyprinid Herpesvirus 2 in Gibel Carp (Carassius auratus gibelio). Antioxidants. 2023;12(2):306-306. doi:https://doi.org/10.3390/antiox12020306

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