Of sick turkeys, kwashiorkor, malaria, perinatal mortality, heroin addicts and food poisoning: research on the influence of aflatoxins on child health in the tropics.

Similarities between the geographical and climatic prevalences of kwashiorkor and of exposure to dietary aflatoxins, and between the biochemical, metabolic and immunological derangements in kwashiorkor and those in animals exposed to aflatoxins, prompted investigation of the associations between kwashiorkor and aflatoxins. Studies in Africa in the 1980s indicated a role for these toxins in the pathogenesis of the disease. Paediatric cases of kwashiorkor are less prone to severe Plasmodium falciparum malaria than normal children. In mice infected with P. berghei, aflatoxin exposure inhibits parasite growth and ameliorates morbidity. Aflatoxins occur in < or = 40% of samples of breast milk from tropical Africa, usually as low concentrations of the relatively non-toxic derivatives of aflatoxin B1 (AFB1) but sometimes as high concentrations of the very toxic AFB1. This could explain kwashiorkor in breast-fed babies. Aflatoxin exposure occurs in > or = 30% of pregnancies in tropical Africa and the toxins are often in cord blood, sometimes at extremely high concentrations. Aflatoxins are now incriminated in neonatal jaundice and there is circumstantial evidence that they cause perinatal death and reduced birthweight. Aflatoxin-induced immunosuppresion may explain the aggressive behaviour of HIV infection in Africa. There are similarities between observations on HIV cases in Africa and those on heroin addicts in Europe, where 'street' heroin is frequently contaminated with aflatoxin. Aflatoxins were found in 20% of random urine samples from heroin addicts in the U.K. and the Netherlands. Aflatoxins have also been incriminated in episodes of food poisoning which have been associated with serious morbidity and mortality, particularly among young children.

Ochratoxin A and aflatoxins in breast milk samples from Sierra Leone.

Breast milk from 113 mothers in two 'Under-Five Clinics' in the Southern Province of Sierra Leone, namely, Njala and Bo, were examined for their mycotoxin content. Only 10 were mycotoxin-free. Eighty-eight per cent of samples contained various aflatoxins and 35% contained ochratoxin A (OTA). Few samples (15%) had a single mycotoxin. Thirty-six (32%) had two mycotoxins and 50 (40%) had three or more. The occurrence of OTA in combination with various aflatoxins was recorded. It is concluded that infants in Sierra Leone are exposed to OTA and aflatoxins at levels which in some cases far exceed those permissible in animal feed in developed countries.

Neonatal jaundice, aflatoxins and naphthols: report of a study in Ibadan, Nigeria.

This study set out to investigate the prevalence of naphthols and aflatoxins in the sera of babies with neonatal jaundice and their mothers in order to determine whether they contribute to the occurrence of unexplained neonatal jaundice in Ibadan. Blood was obtained from 327 jaundiced neonates and 80 of their mothers, and 60 non-jaundiced controls and seven of their mothers admitted to hospital between April 1989 and April 1991. Blood group, bilirubin concentration, erythrocyte G6PD status, aflatoxin and naphthol concentrations in blood were measured. Altogether, 30.9% of the jaundiced neonates were G6PD-deficient, compared with 13.3% of controls (chi 2 = 6.88; p = 0.009). Aflatoxins were detected in 27.4% of jaundiced neonates, 17% of their mothers, 16.6% of controls and 14.4% of control mothers. Naphthols were detected in 7.2% of jaundiced babies, 6.3% of their mothers, 6.25% of control babies and 14.4% of their mothers. Analysis of the data revealed that either G6PD deficiency or the presence of any serum aflatoxin is a risk factor for neonatal jaundice; odds ratio were 2.97 (95%) confidence intervals (CI): 1.31-6.74) and 2.68 (CI: 1.18-6.10), respectively. This study demonstrates that G6PD deficiency and/or the presence of serum aflatoxins are risk factors for neonatal jaundice in Nigeria. Aflatoxins are an additional risk factor not previously reported.

Neonatal jaundice in Zaria, Nigeria--a second prospective study.

Of the 587 neonates born in ABUTH, Zaria, Nigeria and successfully followed up, 99 were clinically jaundiced (16.9%). Of these, only 38 (38%) had significant hyperbilirubinaemia (serum bilirubin above 170 umol/L). During the same period, 279 neonates were admitted through Emergency Paediatric Unit (EPU) of whom 70 (25%) were jaundiced and 64 (95%) of them had serum bilirubin above 170 umol/L. Jaundice was more severe and the incidence of kernicterus higher in babies born outside the hospital than in those born in hospital and periodically followed up. The incidence of kernicterus was 20.3% and 2.6% respectively. The pattern of aetiological factors was similar in the two groups of jaundiced neonates. Septicaemia (50%) and G6PD deficiency (40%) were the major aetiological factors. Exposure to traditional herbal medications, oxytocin induced/augmented labour, cephalhaematoma and tribal incidences did not play statistically significant roles. Jaundice due to Rh-incompatibility was not encountered. Results of this double prospective study were compared with the previous findings in this and other centres in Nigeria.

Neonatal jaundice with reference to aflatoxins: an aetiological study in Zaria, northern Nigeria.

Two prospective studies were undertaken to determine a possible relationship between perinatal aflatoxin exposure and neonatal jaundice. First, cord blood samples from 37 neonates who subsequently developed jaundice and from 40 non-jaundiced (control) babies were analysed for six major aflatoxins and aflatoxicol. Peripheral blood samples of both groups were also analysed postnatally for aflatoxins. In a second study, serum aflatoxin levels of 64 jaundiced neonates admitted from outside the hospital were compared with levels in 60 non-jaundiced control babies. Aflatoxins were detected in 14 (37.8%) cord blood samples of jaundiced neonates and in nine (22.5%) of the controls. The mean cord aflatoxin concentration was highest in jaundiced neonates with septicaemia, but the difference was not statistically significant. The frequency of detection of aflatoxins in peripheral blood was not significantly different in jaundiced and non-jaundiced babies. Aflatoxins were detected in the blood of over 50% of neonates with jaundice of 'unknown' aetiology. There was no correlation between severity of hyperbilirubinaemia and serum aflatoxin levels. Further studies are needed to determine the extent of pre- and postnatal exposure to aflatoxin in Nigerian infants and the effects of such exposure on fetal and neonatal health.

Human fetal exposure to ochratoxin A and aflatoxins.

Analysis of 64 cord blood samples from pregnant women in Sierra Leone revealed the presence of ochratoxin A (OTA) and aflatoxins in 25% and 58% of samples, respectively. Of the eight maternal blood samples collected during delivery, one contained OTA and aflatoxins were detected in six. There was no relationship between mycotoxins in maternal and cord blood. The effect these toxins might have had on the birthweight of infants is discussed.

Ultrastructural changes in murine lymphocytes induced by aflatoxin B1.

This investigation sought to determine whether splenic lymphocytes obtained from Balb/C mice exposed to aflatoxin B1 (AFB1) showed any ultrastructural changes which could account for the immunodysfunction attributable to aflatoxins. Lymphocytes obtained from Balb/C mice administered aflatoxin B1 in olive oil daily for three weeks were studied using both transmission and scanning electron microscopy. The lymphocytes demonstrated ultrastructural changes primarily in the mitochondria where marked internal dissociation of the cristae was revealed by transmission electron microscopy. All other cellular organelles were unaffected. No significant alterations in external structure were observed under scanning electron microscopy. The findings of this study indicate that AFB1 administration does not affect the surface topography of lymphocytes, but AFB1, by causing extensive mitochondrial damage, may affect the way in which these cells function. This could be a possible explanation for the immunodysfunction associated with AFB1.

Detection of naphthols and aflatoxins in Nigerian cord blood.

Widespread use of napthol-containing compounds and frequent contamination of foods by aflatoxins occurs in Nigeria. Napthols cause haemolysis and aflatoxins are hepatotoxic. A study was carried out to determine the extent of fetal exposure to these compounds and their influence on birthweight. Cord blood samples were collected at delivery from 625 babies and their sera were analysed for aflatoxins and naphthols. Mothers' histories and babies' weights were recorded. Naphthols were detected in 6.9% and aflatoxins in 14.6% of serum samples. No correlation was found between the presence of either compound and birthweight. Reported exposure to naphthalene-containing compounds was not related to detection of serum naphthol. Results show considerable fetal exposure to these potentially toxic compounds in Ibadan, Nigeria.

HBsAg and aflatoxins in sera of rural (Igbo-Ora) and urban (Ibadan) populations in Nigeria.

The purpose of this study was to screen for the presence of hepatitis B surface antigen and aflatoxins in the sera of 100 non-hospitalized individuals from the rural population of Igbo-Ora and 89 non-hospitalized individuals from the urban population of Ibadan, Nigeria. Hitherto, such a study as this has not been undertaken in this environment. The proportions of hepatitis B surface antigen carriage and serum 'pathologic' levels of aflatoxins were high (47-49%, 8.2-9.0% respectively) but varied very little between the two different populations sampled. These findings indicate that determined efforts should be instituted to reduce or eliminate hepatitis B virus infection and aflatoxin contamination of high risk foodstuffs from this environment.

HBsAg, aflatoxins and primary hepatocellular carcinoma.

Nigeria is a very high risk area for primary hepatocellular carcinoma and this is the first study to utilize measurements of both hepatitis B virus status and aflatoxin levels in the same patients to determine the role of these factors in the causation of liver cancer in this environment. We have shown that there is a higher prevalence of hepatitis B surface antigen (P < 0.005) and higher 'pathologic' serum levels of aflatoxins (P < 0.05) in patients with primary hepatocellular carcinoma than in matched controls. It is considered that the results of this study may strengthen the hypothesis that hepatitis B virus may be an important aetiological factor in the development of primary hepatocellular carcinoma. Further work is in progress to correlate the level of aflatoxin serum albumin adducts with liver damage in order to assess the value of the albumin adduct as a marker of risk of liver cancer development.

Depression of the Ca(2+)-ATPase activity of the rat liver endoplasmic reticulum by the liver tumour promoters, 1,1,1-trichloro-2,2-bis(p-chlorophenyl)-ethane and phenobarbital.

The effects of a short-term in vivo administration of two liver tumour promoters (phenobarbital and 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane on rat liver endoplasmic reticulum Ca(2+)-ATPase were investigated. The specific activity values of this membrane-bound enzyme significantly decreased (P less than 0.01) by 51% for phenobarbital-treated rats and by 48% for 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane-treated rats compared with control animals. The depression of liver endoplasmic reticulum Ca(2+)-ATPase appears to be a manifestation of the toxicological effect of tumour promoters.

Clinical implications of food contaminated by aflatoxins.

Aflatoxins have been incriminated, mainly on circumstantial evidence, in hepatocellular carcinoma, acute hepatic failure and Reye's syndrome, but other possible effects of continuous or intermittent dietary exposure to aflatoxins, which occurs widely in the tropics, have received little study. Over the past 10 years evidence has steadily accumulated that incriminates aflatoxins in the aetiology of kwashiorkor, a widespread and serious disorder of children in the tropics, previously believed to be caused by protein deficiency. Investigation of human breast milk, undertaken initially to elucidate the pathogenesis of kwashiorkor in breastfed infants, has revealed widespread and serious exposure to aflatoxins from this source. Extension of these studies to pregnant women, in turn, revealed widespread and serious prenatal aflatoxin exposure. In laboratory and farm animals, such exposure has serious implications for immune and hepatic functions, and is detrimental to growth and development. Recent analysis of heroin samples show that heroin addicts may also be exposed to these toxins. These findings show that human exposure to aflatoxins may begin prenatally, persist during breastfeeding, and continue into adult life. It is postulated that aflatoxins (i) play a role in the aetiology of kwashiorkor, (ii) increase neonatal susceptibility to infection and jaundice, (iii) increase childhood susceptibility to infections and malignant disease, (iv) compromise immune responses to prophylactic immunisations and (v) may play a role in the pathogenesis of diseases in heroin addicts. There are indications also that acute, fatal aflatoxin poisoning which masquerades as 'hepatitis' may occur more frequently than is currently appreciated.

Aflatoxin excretion in children with kwashiorkor or marasmic kwashiorkor--a clinical investigation.

A group of five children with kwashiorkor, seven with marasmic kwashiorkor and one underweight child were given an aflatoxin-free diet consisting of maize meal and milk powder. Blood specimens were collected on admission; on day 4 and 10, 24 hour urine and stool samples were collected for the first ten days. Serum, urine and stool samples were analysed for aflatoxins using high performance liquid chromatography with fluorescent detection, after various extraction and clean-up procedures. The children with kwashiorkor and marasmic kwashiorkor excreted aflatoxins in stools for up to 9 and 6 days after admission respectively. No aflatoxins were detected in the stools or urine of the underweight child. In kwashiorkor, urinary excretion ceased after 2 days, while in marasmic kwashiorkor urinary excretion persisted for 4 days. In stools, B1 was the type of aflatoxin detected most frequently in kwashiorkor and least frequently in marasmic kwashiorkor. Aflatoxin M2 was frequently detected in the stools of both groups of children. Estimates of the total amount of aflatoxin excreted by kwashiorkor and marasmic kwashiorkor indicate that these children were harbouring up to 4 micrograms/kg body weight at the time of admission. These findings establish that aflatoxins accumulate in body fluids and tissues in kwashiorkor and marasmic kwashiorkor which is only slowly eliminated.

Foetal and neonatal exposure to aflatoxins.

Studies on 125 primigravidae in rural Kenya revealed aflatoxins in the blood of 54 prenatally. At delivery re-examination of 34 showed aflatoxins in 12 previously negative. The overall detection rate was 53%. Blood from additional 59 women collected at delivery showed aflatoxins in 53%. Aflatoxins were detected in 37% of 101 cord bloods. There was no relationship between aflatoxins in maternal and cord bloods. The frequency of detection was significantly higher in maternal and cord bloods during the 'wet' than 'dry' months. The mean birth weights of females born to aflatoxin positive mothers was significantly lower (255 g) than those born to aflatoxin free mothers. Two stillbirths were recorded, in both cases maternal and cord blood showed aflatoxins. These findings and the adverse effects of prenatal aflatoxin exposure recorded in animal experiments indicate the need for further study of the effects of aflatoxins on the human foetus and newborn.

The effects on malaria of treatment of iron-deficiency anaemia with oral iron in Gambian children.

In order to determine whether giving iron to iron-deficient children increases their susceptibility to malaria, 213 Gambian children aged between 6 months and 5 years with iron-deficiency anaemia were randomized to receive either oral iron or placebo during the rainy season when malaria transmission is maximal. Haematological and iron measurements improved significantly in the group given iron. Regular morbidity surveys showed that fever associated with parasitaemia occurred more frequently in the iron-treated group than in the placebo group. This difference was not significant for all parasitaemias grouped together, but became significant and progressively larger for parasitaemias of ten or more positive fields per 100 high power fields (P less than 0.025), and for parasitaemias of 50 or more positive fields per 100 high power fields (P less than 0.01). Three children in the iron-treated group but none in the placebo group had more than one episode of fever and parasitaemia. Splenomegaly rates rose appreciably during the study in both groups, but in children at age 2 years the splenomegaly rate at the end of the study was significantly greater in the iron-treated group. We concluded that there is a significantly increased risk of fever associated with severe malarial parasitaemia for children with iron-deficiency anaemia given iron during the season of maximal malaria transmission in this part of The Gambia.