Background: Prenatal exposure to lead (Pb) has been shown to have negative and irreversible health impacts on foetal and early childhood development, affecting morbidity and mortality in adulthood. This study aimed to assess in utero Pb exposure, examine birth outcomes, and identify confounding factors in the large cohort of South African population, following the legislated removal of Pb from petrol. Methods: Lead was measured in the maternal blood, urine and cord blood using Inductive Coupled Plasma Mass spectrometry (ICP-MS). The statistical analyses included Spearman’s correlation, Wilcoxon rank sum (Mann Whitney), Kruskal-Wallis rank tests and multivariate linear regression. Results: Overall, the geometric mean (GM) of Pb in maternal blood (PbB) was 1.32 μg/dL (n = 640; 95% CI, 1.24–1.40). In the subset cohort, the GM of paired maternal PbB and cord blood (PbC) was 1.73 μg/dL (n = 350; 95% CI, 1.60–1.86) and 1.26 μg/dL (n = 317; 95% CI, 1.18–1.35), respectively with a positive correlation between the log PbB and the log PbC (rho = 0.65, p = <0.001). Birth outcomes showed geographical differences in the gestational age (p<0.001), birth length (p = 0.028) and head circumference (p<0.001), Apgar score at 5 min (p<0.001) and parity (p<0.002). In female neonates, a positive association was found between PbC and head circumference (rho = 0.243; p<0.016). The maternal PbB levels were positively correlated with race, educational status, water sources, cooking fuels and use of pesticides at home. Conclusions: This study has demonstrated not only the positive impact that the introduction of unleaded petrol and lead-free paint has had on in utero exposure to Pb in South Africa, but has also contributed new data on the topic, in a region where such data and scientific investigations in this field are lacking. Future research should evaluate if similar effects can be detected in young children and the adult population.
Five sites were included in the study: three sites (Sites 1 to 3) were situated in the KwaZulu Natal Province along the Indian Ocean coast (sample collection took place in 2008), and two sites (Sites 4 and 5) were situated in the Western Cape Province along the Atlantic Ocean coast (sample collection took place in 2012 and 2013, respectively (Fig 1). All study sites were rural, except for the urban site of the city of Cape Town (Site 4). The potential candidates recruited for the study were women who were admitted for delivery at the local maternity sections at public hospitals. Women who agreed to participate in the study signed an informed consent form and agreed to donate blood and urine samples before delivery, and to the collection of cord blood samples after delivery. Participants agreed to answer a socio-demographic questionnaire which also included the topics of diet, lifestyle, and self-reported health status. The dietary part of the questionnaire recorded the frequency of intake of various basic foods during pregnancy. Participants also consented to access and use of hospital birth outcome data (including maternal and neonate characteristics such as weight, length and head circumference, gestational age, Apgar score, as well as birth complications, if any) for research purposes. In total, 650 women answered the questionnaire and 640 donated pre-partum blood samples. In a subset cohort of 350 women (from the Indian Ocean site), urine and cord blood samples (n = 317) were also collected. Figure is identical but sites locations were added, and is therefore for representative purposes only. (https://www.cia.gov/library/publications/resources/cia-maps-publications/map-downloads/South%20Africa_Physiography). Ten mL of venous blood was collected before delivery into a BD Vacutainer tube (containing EDTA) and 10 mL of paired cord blood post-partum. Urine (30 mL) was collected before delivery. All samples were stored at -20°C and transported to the South African National Institute for Occupational Health (NIOH) laboratory for analysis. The NIOH participates in the Wadsworth Center–New York State Department of Health Proficiency testing scheme for whole blood and urine. The results obtained are consistently accepted with no indication of bias. The analyses for Pb in the paired maternal and cord whole blood and maternal urine were performed using an Inductively Coupled Plasma-Mass Spectrometer (ICP-MS) instrument (Agilent 7500ce ICP-MS with an Octopole Reaction System). Contamination-free vessels and procedures were used throughout, and validation of results was accomplished by including certified standards, as well as certified reference quality controls, in the analyses. Digested blood samples were analysed for Pb (208) using the no gas acquisition mode and Tl (205) was used as an internal standard. For quality assurance, two certified blood reference controls, viz. Seronorm TM Trace Elements (Sero LTD., Billingstad, Norway) (Levels 1 and 2) were analysed with every analytical run, at intervals between every 10 samples. The detection limit (three times the standard deviation of all blank samples) for Pb in whole blood was 0.04 μg/dL. Acidified urine samples were analysed using the no gas acquisition mode and Au (197) as an internal standard. For urine, five certified reference controls were analysed with every analytical run in intervals of 12 samples (SeronormTM Trace Elements in urine (urine blank), Lot: OK4636; SeronormTM Trace Elements in urine, Lot: 0511545; UTAK Urine control, Lot: 1170; Lyphochek Urine Metals Control level 1, Lot: 69121; Lyphochek Urine Metals Control level 2, Lot: 69122). The detection limit (three times the standard deviation of all blank samples) for Pb in urine was 0.52 μg/L. Covariate information was obtained during the questionnaire-based interview and from medical records. Maternal weight and height were recorded at the hospital on admission. From the medical records, the following neonate characteristics were retrieved: birth weight (grams), birth length (cm), head circumference (cm) and gestational age (weeks). Pre-term labour was defined as mothers giving birth at less than 37 weeks gestational age. Education was categorised as no education to completed primary school, completed secondary school and any level of tertiary education reached. Maternal tobacco smoking during pregnancy was defined as yes or no. Exposure to environmental tobacco smoke (ETS) was defined as exposure to tobacco smoke from smoking by others in the household. A binary classification was used for exposure to indoor smoke from the burning of fossil fuel (wood and coal) for the purpose of heating or cooking, separating study participants into those exposed to fossil fuel and those not exposed (for example, those using electricity). Dietary questions relating to the intake of proteins, carbohydrates, dairy products, tea, coffee, bottled water, fruits, as well as vine, root and leafy vegetables, were assessed and classified as daily, at least once a week and seldom (both for pre-pregnancy and during pregnancy). The statistical analyses were performed using STATA (StataCorp, 2013. Stata Statistical Software: Release 13. College Station, TX: StataCorp LP). Pb was detected in all blood and urine samples. Bivariate analyses between maternal PbB exposure and covariates were evaluated by Spearman’s correlation coefficient, Wilcoxon rank sum (Mann Whitney), Kruskal-Wallis rank tests and linear regression as appropriate. The distribution of Pb levels in maternal blood and in cord blood, were skewed and were log transformed. Multivariate linear regression was carried out using a backward deletion approach, starting with a full model of factors significantly associated with natural logarithm-transformed maternal Pb levels in the bivariate analysis. All statistical tests were two tailed and statistical significance was set at p < 0.05. Ethics approval for the study was obtained from the Human Research Ethics Committee of the University of Witwatersrand in Johannesburg (Protocol no. {"type":"entrez-nucleotide","attrs":{"text":"M10742","term_id":"147973","term_text":"M10742"}}M10742), and from the relevant provincial Departments of Health: the Provincial Department of Health of KwaZulu Natal and the Western Cape Provincial Department of Health. In addition, CEOs of the respective hospitals had to confirm that he/she allowed the research work to proceed. Identical procedures were followed in terms of obtaining consent from participants. Confidentiality was maintained by assigning identification numbers to all study participants. During the informed consent process, it was emphasised that participation was voluntary and could be withdrawn at any time.