Aflatoxin Exposure during Early Life Is Associated with Differential DNA Methylation in Two-Year-Old Gambian Children.

Background: DNA methylation is an epigenetic control mechanism that may be altered by environmental exposures. We have previously reported that in utero exposure to the mycotoxin and liver carcinogen aflatoxin B1 from the maternal diet, as measured using biomarkers in the mothers' blood, was associated with differential DNA methylation in white blood cells of 6-month-old infants from The Gambia. Methods: Here we examined aflatoxin B1-associated differential DNA methylation in white blood cells of 24-month-old children from the same population (n = 244), in relation to the child's dietary exposure assessed using aflatoxin albumin biomarkers in blood samples collected at 6, 12 and 18 months of age. HM450 BeadChip arrays were used to assess DNA methylation, with data compared to aflatoxin albumin adduct levels using two approaches; a continuous model comparing aflatoxin adducts measured in samples collected at 18 months to DNA methylation at 24 months, and a categorical time-dose model that took into account aflatoxin adduct levels at 6, 12 and 18 months, for comparison to DNA methylation at 24 months. Results: Geometric mean (95% confidence intervals) for aflatoxin albumin levels were 3.78 (3.29, 4.34) at 6 months, 25.1 (21.67, 29.13) at 12 months and 49.48 (43.34, 56.49) at 18 months of age. A number of differentially methylated CpG positions and regions were associated with aflatoxin exposure, some of which affected gene expression. Pathway analysis highlighted effects on genes involved with with inflammatory, signalling and growth pathways. Conclusions: This study provides further evidence that exposure to aflatoxin in early childhood may impact on DNA methylation.

Estimating the health burden of aflatoxin attributable stunting among children in low income countries of Africa.

Numerous population-based studies have documented high prevalence of aflatoxin associated childhood stunting in low income countries. We provide an estimate of the disease burden of aflatoxin related stunting using data from the four African countries. For this empirical analysis, we obtained blood aflatoxin albumin adduct biomarker based exposure data as measured using ELISA technique and anthropometric measurement data from surveys done over a 12-year period from 2001 to 2012 in four low income countries in Africa. We used these data to calculate population attributable risk (PAR), life time disease burden for children under five by comparing two groups of stunted children using both prevalence and incidence-based approaches. We combined prevalence estimates with a disability weight, measuring childhood stunting and co-occurrence of stunting-underweight to produce years lived with disability. Using a previously reported mortality, years of life lost were estimated. We used probabilistic analysis to model these associations to estimate the disability-adjusted life-years (DALYs), and compared these with those given by the Institute for Health Metrics and Evaluation's Global Burden of Disease (GBD) 2016 study. The PAR increased from 3 to 36% for aflatoxin-related stunting and 14-50% for co-occurrence of stunting and underweight. Using prevalence-based approach, children with aflatoxin related stunting resulted in 48,965.20 (95% uncertainty interval (UI): 45,868.75-52,207.53) DALYs per 100,000 individuals. Children with co-occurrence of stunting and underweight due to exposure to aflatoxin resulted in 40,703.41 (95% UI: 38,041.57-43,517.89) DALYs per 100,000 individuals. Uncertainty analysis revealed that reducing aflatoxin exposure in high exposure areas upto non-detectable levels could save the stunting DALYs up to 50%. The burden of childhood all causes stunting is greater in countries with higher aflatoxin exposure such as Benin. In high exposure areas, these results might help guide research protocols and prioritisation efforts and focus aflatoxin exposure reduction. HEFCE Global Challenge Research Fund Aflatoxin project.

Aflatoxin exposure in pregnant women of mixed status of human immunodeficiency virus infection and rate of gestational weight gain: a Ugandan cohort study.

OBJECTIVES: To examine the association between aflatoxin (AF) exposure during pregnancy and rate of gestational weight gain (GWG) in a sample of pregnant women of mixed HIV status in Gulu, northern Uganda. METHODS: 403 pregnant women were included (133 HIV-infected on antiretroviral therapy (ART), 270 HIV-uninfected). Women's weight, height and socio-demographic characteristics were collected at baseline (~19 weeks' gestation); weight was assessed at each follow-up visit. Serum was collected at baseline and tested for aflatoxin B1 -lysine adduct (AFB-lys) levels using high-performance liquid chromatography (HPLC). Linear mixed-effects models were used to examine the association between AFB-lys levels and rate of GWG. RESULTS: AFB-lys levels (detected in 98.3% of samples) were higher among HIV-infected pregnant women than HIV-uninfected pregnant women [median (interquartile range): 4.8 (2.0, 15.0) vs. 3.5 (1.6, 6.1) pg/mg of albumin, P < 0.0001]. Adjusting for HIV status, a one-log increase in aflatoxin levels was associated with a 16.2 g per week lower rate of GWG (P = 0.028). The association between AFB-lys and the rate of GWG was stronger and significant only among HIV-infected women on ART [-25.7 g per week per log (AFB-lys), P = 0.009 for HIV-infected women vs. -7.5 g per week per log (AFB-lys), P = 0.422 for HIV-uninfected women]. CONCLUSIONS: Pregnant women with higher levels of AF exposure had lower rates of GWG. The association was stronger for HIV-infected women on ART, suggesting increased risk.

OBJECTIFS: Examiner l'association entre l'exposition à l'aflatoxine (AF) pendant la grossesse et le taux de gain de poids pendant la grossesse (GWG) dans un échantillon de femmes enceintes de statut mixte VIH à Gulu, dans le nord de l' Ouganda. MÉTHODES: 403 femmes enceintes ont été incluses (133 infectées par le VIH sous traitement antirétroviral (ART), 270 non infectées par le VIH). Le poids, la taille et les caractéristiques sociodémographiques des femmes ont été collectés au départ (~19 semaines de gestation); le poids a été évalué à chaque visite de suivi. Le sérum a été recueilli au départ et testé pour les niveaux d'adduit d'aflatoxine B1 -lysine (AFB-lys) en utilisant la chromatographie liquide à haute performance (HPLC). Des modèles linéaires d'effets mixtes ont été utilisés pour examiner l'association entre les niveaux de AFB-lys et le taux de GWG. RÉSULTATS: Les niveaux de AFB-lys (détectés dans 98,3% des échantillons) étaient plus élevés chez les femmes enceintes infectées par le VIH que chez celles enceintes non infectées par le VIH [médiane (intervalle interquartile): 4,8 (2,0, 15,0) vs 3,5 (1,6, 6,1) pg/mg d'albumine, P <0,0001]. En ajustant pour le statut VIH, une augmentation d'un log des niveaux d'aflatoxine était associée à un taux de GWG inférieur de 16,2 g par semaine (P = 0,028). L'association entre AFB-lys et le taux de GWG était plus forte et significative seulement chez les femmes infectées par le VIH sous ART [-25,7 g par semaine et par log (AFB-lys), P = 0,009 pour les femmes infectées par le VIH contre -7,5 g par semaine et par log (AFB-lys), P = 0,422 pour les femmes non infectées par le VIH]. CONCLUSIONS: Les femmes enceintes présentant des niveaux plus élevés d'exposition à l'AF avaient des taux de GWG plus faibles. L'association était plus forte pour les femmes infectées par le VIH sous ART, ce qui suggère un risque accru.

Corn Flour Intake, Aflatoxin B1 Exposure, and Risk of Esophageal Precancerous Lesions in a High-Risk Area of Huai'an, China: A Case-Control Study.

Aflatoxin B1 (AFB1), which has potent toxicity and carcinogenicity, is a common contaminant of important agricultural commodities. This study aimed to investigate the frequency of corn flour intake and assess the exposure to AFB1 via direct detection of AFB1 in the diet and serum AFB1 exposure biomarker, so as to evaluate their associations with the risk of esophageal precancerous lesions (EPL). A case-control study based on three-day duplicate diet samples was performed in Huai'an District. One hundred EPL cases and 100 healthy controls were enrolled and required to be age- (±2 years) and gender-matched. The concentration of AFB1 in food samples and the level of serum AFB1-albumin (AFB1-Alb) adduct were quantitatively analyzed. Results showed that corn flour intake was positively associated with serum AFB1-Alb adduct level (p for trend = 0.003), dietary AFB1 exposure (p for trend < 0.001), and the risk of EPL (p for trend = 0.017). Increased serum AFB1-Alb adduct level was associated with an increased risk of EPL as well (p for trend < 0.001). In conclusion, corn flour may be an essential source of AFB1 in Huai'an District, whereas high exposure to AFB1 is likely to be an important risk factor contributing to the progression of EPL.

Preliminary study on the relationship between aflatoxin-bovine serum albumin adducts in blood and aflatoxin M1 levels in milk of dairy cows.

The aflatoxin (AF) albumin adduct is often used as a biomarker for aflatoxin exposure in humans. An ELISA method previously used for aflatoxin serum albumin in human blood was used to analyse bovine serum samples (n = 22) collected from dairy cattle during an aflatoxin mitigation study in Kenya. Albumin adduct data were compared with aflatoxin M1 (AFM1) levels in corresponding milk samples from these cows. The concentration ranged from < LOD to 487.9 pg/mL for AFM1 and < LOD and 96.3 pg/mg for aflatoxin-albumin. This study indicates that aflatoxin-albumin adducts could be used as a measure of chronic aflatoxin exposure in dairy cattle.

Mycotoxin exposure is associated with increased risk of esophageal squamous cell carcinoma in Huaian area, China.

BACKGROUND: Consumption of moldy food has previously been identified as a risk factor for esophageal squamous cell carcinoma (ESCC) in high-risk countries; however, what contributing roles these dietary carcinogenic mycotoxins play in the etiology of ESCC are largely unknown. METHODS: A mycotoxin biomarker-incorporated, population-based case-control study was performed in Huaian area, Jiangsu Province, one of the two high-risk areas in China. Exposure biomarkers of aflatoxins (AF) and fumonisins (FN) were quantitatively analyzed using HPLC-fluorescence techniques. RESULTS: Among the cases (n = 190), the median levels of AF biomarker, serum AFB1-lysine adduct, and FN biomarker, urinary FB1, were 1.77 pg/mg albumin and 176.13 pg/mg creatinine, respectively. Among the controls (n = 380), the median levels of AFB1-lysine adduct and urinary FB1 were 1.49 pg/mg albumin and 56.92 pg/mg creatinine, respectively. These mycotoxin exposure biomarker levels were significantly higher in cases as compared to controls (p <  0.05 and 0.01, respectively). An increased risk to ESCC was associated with exposure to both AFB1 and FB1 (p <  0.001 for both). CONCLUSIONS: Mycotoxin exposure, especially to AFB1 and FB1, was associated with the risk of ESCC, and a greater-than-additive interaction between co-exposures to these two mycotoxins may contribute to the increased risk of ESCC in Huaian area, China.

Aflatoxin levels in maize and peanut and blood in women and children: The case of Timor-Leste.

Aflatoxins are toxic fungal metabolites produced by Aspergillus sp. with carcinogenic properties that are a common food contaminant of many crops including maize and peanuts. In Timor-Leste malnutrition and children's stunting are frequent and maize and peanuts are staple foods. This study aimed to provide information on aflatoxin exposure nationally. The study measured levels of aflatoxin in locally-produced maize and peanuts (296 samples) and of aflatoxin-albumin conjugate in blood samples of women and young children (514 and 620 respectively) across all municipalities. The average concentration of aflatoxin in the grain samples was low with most maize (88%) and peanut (92%) samples - lower than European Commission tolerated aflatoxin level. Although aflatoxin-albumin conjugate was detected in more than 80% of blood samples, the average concentration in children and adults of 0.64 and 0.98 pg mg-1 alb, respectively, is much lower than in other similar rural-based countries. Although low in concentration, blood aflatoxin levels and aflatoxin contamination levels in maize across municipalities were correlated significantly for mothers (R2 = 37%, n = 495) but not for children (R2 = 10%). It is unlikely that the consumption of aflatoxin contaminated grain is a causative factor in the current level of malnutrition and stunting affecting Timor-Leste children.

Biomonitoring Data for Assessing Aflatoxins and Ochratoxin A Exposure by Italian Feedstuffs Workers.

Mycotoxins exposure by inhalation and/or dermal contact is possible in different branches of industry especially where heavily dusty settings are present and the handling of dusty commodities is performed. This study aims to explore the validity of the biomonitoring as a tool to investigate the intake of mycotoxins in a population of workers operating in an Italian feed plant. Serum samples were collected for the determination of aflatoxins B1 (AFB1), AFB1-Lysine adduct and ochratoxin A (OTA). A method based on liquid-liquid extraction coupled with high resolution mass spectrometry determination was developed and fully validated. For AFB1, a high number of non-detected samples (90%) was found and no statistical difference was observed comparing workers and control group. None of the analyzed samples showed the presence of AFB1-Lysine adduct. For OTA, the 100% of the analyzed samples was positive with a 33% of the samples showing a concentration higher than the limit of quantification (LOQ), but no statistical difference was highlighted between the average levels of exposed and control groups. In conclusion, the presence of AFB1 and OTA in serum cannot be attributable to occupational exposure.

The effects of aflatoxin exposure on Hepatitis B-vaccine induced immunity in Kenyan children.

BACKGROUND: Globally, approximately three million children die each year from vaccine preventable infectious diseases mainly in developing countries. Despite the success of the expanded immunization program, not all infants and children around the world develop the same protective immune response to the same vaccine. A vaccine must induce a response over the basal immune response that may be driven by population-specific, environmental or socio-economic factors. Mycotoxins like aflatoxins are immune suppressants that are confirmed to interfere with both cell-mediated and acquired immunity. The mechanism of aflatoxin toxicity is through the binding of the bio-activated AFB1-8, 9-epoxide to cellular macromolecules. METHODS: We studied Hepatitis B surface antibodies [anti-HBs] levels to explore the immune modulation effects of dietary exposure to aflatoxins in children aged between one and fourteen years in Kenya. Hepatitis B vaccine was introduced for routine administration for Kenyan infants in November 2001. To assess the effects of aflatoxin on immunogenicity of childhood vaccines Aflatoxin B1-lysine in blood serum samples were determined using High Performance Liquid Chromatography with Fluorescence detection while anti-HBs were measured using Bio-ELISA anti-HBs kit. RESULTS: The mean ± SD of AFB1-lysine adducts in our study population was 45.38 ± 87.03 pg/mg of albumin while the geometric mean was 20.40 pg/mg. The distribution of AFB1-lysine adducts was skewed to the right. Only 98/205 (47.8%) of the study population tested positive for Hepatitis B surface antibodies. From regression analysis, we noted that for every unit rise in serum aflatoxin level, anti-HBs dropped by 0.91 mIU/ml (-0.9110038; 95% C.I -1.604948, -0.21706). CONCLUSION: Despite high coverage of routine immunization, less than half of the study population had developed immunity to HepB. Exposure to aflatoxin was high and weakly associated with low anti-HBs antibodies. These findings highlight a potentially significant role for environmental factors that may contribute to vaccine effectiveness warranting further research.

Geospatial visualisation of food contaminant distributions: Polychlorinated naphthalenes (PCNs), potentially toxic elements (PTEs) and aflatoxins.

Large volume of multidimensional data can be summarised, both in terms of tabulated statistics, and as graphic geospatial visualisations. The latter approach allows rapid interpretation and communication of complex information to stake-holders such as regulators, risk assessors and policy makers. In the main study on polychlorinated naphthalene (PCN), individual samples representing different edible fish species were analysed from around the UK. PCNs were observed in all samples with nearly all of the twelve measured congeners being detected. Summed congener concentrations ranged from 0.7 ng/kg ww (turbot) to 265 ng/kg ww (sprats). The highest contamination levels were recorded for sprats and mackerel with mean summed concentrations of 67 ng/kg ww and 68 ng/kg ww respectively. Two ancillary studies, on potentially toxic elements (PTEs) in crabs from China and aflatoxin in children's blood from Tanzania, demonstrate the wide applicability of this approach. The PTE contents in crab showed strong dependence on the tested tissues and elements, and crabs from Tai and Yangcheng Lakes showed obviously higher PTE levels than the other lakes. Geospatial distribution of the aflatoxin biomarker AF-alb in children's serum from 3 locations showed how individual anthropometric or socio-economic data reveals the relationship between family size, socio-economic score and magnitude of serum aflatoxin levels. In addition to facilitating the flow of interpreted data to stakeholders, these techniques can direct the formulation of risk mitigation activities and help with the identification of data gaps. When combined with hierarchical cluster analyses, correlations within the data can also be predicted.

Antibody-functionalized reduced graphene oxide films for highly selective capture and purification of aflatoxins.

Pyrenylbutyric acid and streptavidin were coupled to films of reduced graphene oxide (rGO) and then conjugated to a biotinylated broad-spectrum monoclonal antibody against aflatoxins (AFs). It is shown that such films can efficiently and selectively capture AFs inculding AFB1, AFB2, AFG1, AFG2, AFM1 and AFM2. The rGO films were characterized by using scanning electron microscopy, energy-dispersive spectroscopy, and raman spectroscopy. The selectivity and purification performance of the antibody-loaded rGO films were investigated. They were applied to the purification of extremely small samples (100 µL) of AFs-spiked rabbit serum after enzymatic hydrolysis. The AFs were analyzed by ultra-performance liquid chromatography coupled to tandem mass spectrometry. The limits of detection for the six AFs investigated ranged from 50 to 170 pg·mL-1. The average recoveries of AFs in spiked rabbit serum samples ranged from 55% to 75%, with relative standard deviations of less than 9.4%. Graphical abstract Design of a multifunctional sandwich film that consists of a reduced graphene oxide film base, a pyrenylbutyric acid middle layer and a broad-specificity anti-AF monoclonal antibody surface layer. It was successfully applied to the determination of aflatoxins in only 100 µL of rabbit serum samples with satisfactory selectivity and acceptable accuracy.

Impaired growth in rural Gambian infants exposed to aflatoxin: a prospective cohort study.

BACKGROUND: Exposure to aflatoxin, a mycotoxin produced by fungi that commonly contaminates cereal crops across sub-Saharan Africa, has been associated with impaired child growth. We investigated the impact of aflatoxin exposure on the growth of Gambian infants from birth to two years of age, and the impact on insulin-like growth factor (IGF)-axis proteins. METHODS: A subsample (N = 374) of infants from the Early Nutrition and Immune Development (ENID) trial (ISRCTN49285450) were included in this study. Aflatoxin-albumin adducts (AF-alb) were measured in blood collected from infants at 6, 12 and 18 months of age. IGF-1 and IGFBP-3 were measured in blood collected at 12 and 18 months. Anthropometric measurements taken at 6, 12, 18 and 24 months of age were converted to z-scores against the WHO reference. The relationship between aflatoxin exposure and growth was analysed using multi-level modelling. RESULTS: Inverse relationships were observed between lnAF-alb and length-for-age (LAZ), weight-for-age (WAZ), and weight-for-length (WLZ) z-scores from 6 to 18 months of age (ß = - 0·04, P = 0·015; ß = - 0·05, P = 0.003; ß = - 0·06, P = 0·007; respectively). There was an inverse relationship between lnAF-alb at 6 months and change in WLZ between 6 and 12 months (ß = - 0·01; P = 0·013). LnAF-alb at 12 months was associated with changes in LAZ and infant length between 12 and 18 months of age (ß = - 0·01, P = 0·003; ß = - 0·003, P = 0·02; respectively). LnAF-alb at 6 months was associated with IGFBP-3 at 12 months (r = - 0·12; P = 0·043). CONCLUSIONS: This study found a small but significant effect of aflatoxin exposure on the growth of Gambian infants. This relationship is not apparently explained by aflatoxin induced changes in the IGF-axis.

Comparison of urinary aflatoxin M1 and aflatoxin albumin adducts as biomarkers for assessing aflatoxin exposure in Tanzanian children.

PURPOSE: To determine levels of urinary aflatoxin M1 (AFM1) in children and correlate the concentrations with previously reported aflatoxin albumin adduct (AF-alb) levels in these children. MATERIALS AND METHODS: Matched urine and blood samples were collected from 84 Tanzanian children aged 6-14 months old. From 31 children in one village (Kigwa), samples were collected at three time points six months apart. Samples were collected from 31 and 22 children from two different regions at the second time point only. Urinary AFM1 was measured using a commercial enzyme-linked immunosorbent assay (ELISA) kit with a modified protocol to improve sensitivity. AF-alb was measured using an established ELISA method. RESULTS: The relative ranking of the three villages for exposure to aflatoxin based on either AFM1 or AF-alb biomarker measurements was the same. In Kigwa village, both AFM1 and AF-alb levels were higher at six months post-harvest compared to baseline. However, at the next visit, the AFM1 levels dropped from a GM (interquartile range) of 71.0 (44.7, 112.6) at visit two to 49.3 (31.5, 77.3) pg/ml urine, whereas AF-alb levels increased from 47.3 (29.7, 75.2) to 52.7 (35.4, 78.3) pg/mg albumin between these two visits, reflecting the fact that AFM1 measures short-term exposure, whereas AF-alb measures longer term exposure. There was a correlation between AFB1 intake and AFM1 excretion (r= 0.442, p ≤ 0.001). CONCLUSIONS: Urinary AFM1 is a good biomarker for AFB1 exposure in Tanzanian children, reflecting geographical and temporal variations in exposure to this foodborne toxin.

Aflatoxin and viral hepatitis exposures in Guatemala: Molecular biomarkers reveal a unique profile of risk factors in a region of high liver cancer incidence.

Liver cancer is an emerging global health issue, with rising incidence in both the United States and the economically developing world. Although Guatemala experiences the highest rates of this disease in the Western hemisphere and a unique 1:1 distribution in men and women, few studies have focused on this population. Thus, we determined the prevalence and correlates of aflatoxin B1 (AFB1) exposure and hepatitis virus infection in Guatemalan adults. Healthy men and women aged ≥40 years (n = 461), residing in five departments of Guatemala, were enrolled in a cross-sectional study from May-October of 2016. Serum AFB1-albumin adducts were quantified using isotope dilution mass spectrometry. Multivariate linear regression was used to assess relationships between AFB1-albumin adduct levels and demographic factors. Biomarkers of hepatitis B virus and hepatitis C virus infection were assessed by immunoassay and analyzed by Fisher's exact test. AFB1-albumin adducts were detected in 100% of participants, with a median of 8.4 pg/mg albumin (range, 0.2-814.8). Exposure was significantly higher (p<0.05) in male, rural, low-income, and less-educated participants than in female, urban, and higher socioeconomic status participants. Hepatitis B and C seropositivity was low (0.9% and 0.5%, respectively). Substantial AFB1 exposure exists in Guatemalan adults, concurrent with low prevalence of hepatitis virus seropositivity. Quantitatively, AFB1 exposures are similar to those previously found to increase risk for liver cancer in Asia and Africa. Mitigation of AFB1 exposure may reduce liver cancer incidence and mortality in Guatemala, warranting further investigation.

Survey on Urinary Levels of Aflatoxins in Professionally Exposed Workers.

Feed mill workers may handle or process maize contaminated with aflatoxins (AFs). This condition may lead to an unacceptable intake of toxins deriving from occupational exposure. This study assessed the serological and urinary levels of AFs in workers exposed to potentially contaminated dusts in two mills. From March to April 2014, blood and urine samples were collected, on Monday and Friday morning of the same working week from 29 exposed workers and 30 non-exposed controls. AFs (M1, G2, G1, B1, B2) and aflatoxicol (AFOH) A were analyzed. Each subject filled in a questionnaire to evaluate potential food-borne exposures to mycotoxins. AFs contamination in environmental dust was measured in both plants. No serum sample was found to be positive. Seventy four percent of urine samples (73.7%) revealed AFM1 presence. AFM1 mean concentration was 0.035 and 0.027 ng/mL in exposed and non-exposed workers, respectively (p = 0.432); the concentration was slightly higher in Friday's than in Monday's samples, in exposed workers, 0.040 versus (vs.) 0.031 and non-exposed controls (0.030 vs. 0.024, p = 0.437). Environmental AFs contamination ranged from 7.2 to 125.4 µg/kg. The findings of this study reveal the presence of higher AFs concentration in exposed workers than in non-exposed controls, although these differences are to be considered consistent with random fluctuations.

Chronic aflatoxin exposure in children living in Bhaktapur, Nepal: Extension of the MAL-ED study.

Exposure to aflatoxin, a mycotoxin common in maize and groundnuts, has been associated with childhood stunting in sub-Saharan Africa. In an effort to further our understanding of growth impairment in relation to mycotoxins and other risk factors, biospecimens from a cohort of children enrolled in the Bhaktapur, Nepal MAL-ED study were assessed for aflatoxin exposure at 15, 24, and 36 months of age. Exposure was assessed through a well-established serum biomarker, the AFB1-lysine adduct. In this manuscript, the levels of aflatoxin exposure in the Nepal cohort were compared with those observed in aflatoxin studies, with child growth parameters as a health outcome. Results from this preliminary analysis demonstrated chronic aflatoxin exposure in children residing in Bhaktapur with a geometric mean of 3.62 pg AFB1-lysine/mg albumin. The range of exposure in this population is similar to those in African populations where associations with aflatoxin biomarkers and poor child growth have been observed. Future work will analyze the relationships between aflatoxin levels, growth, and other risk factors collected by the MAL-ED study.

Dietary exposure to aflatoxin and micronutrient status among young children from Guinea.

SCOPE: Aflatoxin exposure coincides with micronutrient deficiencies in developing countries. Animal feeding studies have postulated that aflatoxin exposure may be exacerbating micronutrient deficiencies. Evidence available in human subjects is limited and inconsistent. The aim of the study was to investigate the relationship between aflatoxin exposure and micronutrient status among young Guinean children. METHODS AND RESULTS: A total of 305 children (28.8 ± 8.4 months) were recruited at groundnut harvest (rainy season), of which 288 were followed up 6 months later postharvest (dry season). Blood samples were collected at each visit. Aflatoxin-albumin adduct levels were measured by ELISA. Vitamin A, vitamin E and ß-carotene concentrations were measured using HPLC methods. Zinc was measured by atomic absorption spectroscopy. Aflatoxin exposure and micronutrient deficiencies were prevalent in this population and were influenced by season, with levels increasing between harvest and postharvest. At harvest, children in the highest aflatoxin exposure group, compared to the lowest, were 1.98 (95%CI: 1.00, 3.92) and 3.56 (95%CI: 1.13, 11.15) times more likely to be zinc and vitamin A deficient. CONCLUSION: Although children with high aflatoxin exposure levels were more likely to be zinc and vitamin A deficient, further research is necessary to determine a cause and effect relationship.

Temporal Variation and Association of Aflatoxin B1 Albumin-Adduct Levels with Socio-Economic and Food Consumption Factors in HIV Positive Adults.

The association between aflatoxin exposure and alteration in immune responses observed in humans suggest that aflatoxin could suppress the immune system and work synergistically with HIV to increase disease severity and progression to AIDS. No longitudinal study has been conducted to assess exposure to aflatoxin (AF) among HIV positive individuals. We examined temporal variation in AFB1 albumin adducts (AF-ALB) in HIV positive Ghanaians, and assessed the association with socioeconomic and food consumption factors. We collected socioeconomic and food consumption data for 307 HIV positive antiretroviral naive adults and examined AF-ALB levels at recruitment (baseline) and at six (follow-up 1) and 12 (follow-up 2) months post-recruitment, by age, gender, socioeconomic status (SES) and food consumption patterns. Generalized linear models were used to examine the influence of socioeconomic and food consumption factors on changes in AF-ALB levels over the study period, adjusting for other covariates. AF-ALB levels (pg/mg albumin) were lower at baseline (mean AF-ALB: 14.9, SD: 15.9), higher at six months (mean AF-ALB: 23.3, SD: 26.6), and lower at 12 months (mean AF-ALB: 15.3, SD: 15.4). Participants with the lowest SES had the highest AF-ALB levels at baseline and follow up-2 compared with those with higher SES. Participants who bought less than 20% of their food and who stored maize for less than two months had lower AF-ALB levels. In the adjusted models, there was a statistically significant association between follow up time and season (dry or rainy season) on AF-ALB levels over time (p = 0.04). Asymptomatic HIV-positive Ghanaians had high plasma AF-ALB levels that varied according to season, socioeconomic status, and food consumption patterns. Steps need to be taken to ensure the safety and security of the food supply for the population, but in particular for the most vulnerable groups such as HIV positive people.

Exposure to aflatoxin B1 in utero is associated with DNA methylation in white blood cells of infants in The Gambia.

BACKGROUND: Exposure to environmental toxins during embryonic development may lead to epigenetic changes that influence disease risk in later life. Aflatoxin is a contaminant of staple foods in sub-Saharan Africa, is a known human liver carcinogen and has been associated with stunting in infants. METHODS: We have measured aflatoxin exposure in 115 pregnant women in The Gambia and examined the DNA methylation status of white blood cells from their infants at 2-8 months old (mean 3.6 ± 0.9). Aflatoxin exposure in women was assessed using an ELISA method to measure aflatoxin albumin (AF-alb) adducts in plasma taken at 1-16 weeks of pregnancy. Genome-wide DNA methylation of infant white blood cells was measured using the Illumina Infinium HumanMethylation450beadchip. RESULTS: AF-alb levels ranged from 3.9 to 458.4 pg/mg albumin. We found that aflatoxin exposure in the mothers was associated to DNA methylation in their infants for 71 CpG sites (false discovery rate < 0.05), with an average effect size of 1.7% change in methylation. Aflatoxin-associated differential methylation was observed in growth factor genes such as FGF12 and IGF1, and immune-related genes such as CCL28, TLR2 and TGFBI. Moreover, one aflatoxin-associated methylation region (corresponding to the miR-4520b locus) was identified. CONCLUSIONS: This study shows that maternal exposure to aflatoxin during the early stages of pregnancy is associated with differential DNA methylation patterns of infants, including in genes related to growth and immune function. This reinforces the need for interventions to reduce aflatoxin exposure, especially during critical periods of fetal and infant development.