Metals and arsenic distribution in stray dogs' tissues around a lead-zinc mine in Kabwe, Zambia.

Metal contamination poses a threat to human, animal, and environmental health. The purpose of the current study was to assess the accumulation of toxic and trace metal concentrations in tissues of stray dogs in Kabwe, a town known for lead pollution due to a long history of lead and zinc mining. Brain, spleen, heart, stomach, stomach content, small intestine, kidney, liver, and bone samples were collected from 29 stray dogs (14 from locations within 3 km of the mine and 15 from sites 7 km away from the mine) after necropsy. Inductively coupled plasma mass spectrometry was used to analyze toxic metals, arsenic (As), cadmium (Cd), and lead (Pb), and trace elements, copper (Cu) and zinc (Zn). The lungs (0.117 ± 0.114 mg/kg dry weight), kidney (7.515 ± 8.830 mg/kg dry weight), and bone (41.68 ± 66.83 mg/kg dry weight) were found to have the highest concentrations of As, Cd, and Pb, respectively. In contrast, higher Cu and Zn concentrations were measured in the liver than in other tissues. In all tissues analyzed, tissues sampled from dogs near the mine had significantly higher mean concentrations of Cd and Pb than dogs far away. Neither sex nor age-related differences were observed in the distribution of metals in most tissues. There were significant associations among toxic (Pb and Cd) and trace metals (Cu and Zn). In the kidney, Cd positively correlated with Pb (ρ = 0.534) and Zn (ρ = 0.600), whereas in the liver, Cu correlated with Zn (ρ = 0.565). The current study's findings suggest that environmental pollution is still a problem in Kabwe, and environmental remediation is needed to address the pollution.

Persistent organic pollutants (POPs) and per- and polyfluoroalkyl substances (PFASs) in liver from wild and farmed tilapia (Oreochromis niloticus) from Lake Kariba, Zambia: Levels and geographic trends and considerations in relation to environmental quality standards (EQSs).

The current study was carried out to investigate a wide variety of persistent organic pollutants (POPs) in wild and farmed tilapia (Oreochromis niloticus) in Lake Kariba, Zambia, and assess levels of POPs in relation to Environmental Quality Standards (EQSs). Concentrations of organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), polybrominated diphenyls (PBDEs), and perfluoroalkyl substances (PFASs) were determined in liver samples of tilapia. PFASs compounds PFOS, PFDA and PFNA were only detected in wild fish, with the highest median PFOS levels in site 1 (0.66 ng/g ww). Concentrations of POPs were in general highest in wild tilapia. The highest median ∑DDTs (93 and 81 ng/g lw) were found in wild tilapia from sites 1 and 2, respectively 165 km and 100 km west of the fish farms. Lower DDE/DDT ratios in sites 1 and 3 may indicate relatively recent exposure to DDT. The highest median of ∑17PCBs (3.2 ng/g lw) and ∑10PBDEs (8.1 ng/g lw) were found in wild tilapia from sites 1 and 2, respectively. The dominating PCB congeners were PCB-118, -138, -153 and -180 and for PBDEs, BDE-47, -154, and -209. In 78% of wild fish and 8% of farmed fish ∑6PBDE concentrations were above EQSbiota limits set by the EU. This warrants further studies.

Association between Chronic Environmental Lead (Pb) Exposure and Cytokines in Males and Females of Reproductive Age from Kabwe, Zambia.

Lead (Pb) poisoning remains a great public health challenge globally known to induce a wide range of ailments in both children and adults. The current study investigated the association of chronic environmental Pb exposure and immunomodulatory cytokines tumor necrosis factor alpha (TNF-α) and interleukin-8 (IL-8) in adult males and females living in Kabwe, Zambia. The standard human cytokine/chemokine Milliplex assay was used to quantify plasma cytokines from four groups categorized as low (<10 µg/dL) and high (>10 µg/dL) blood lead level (BLL) groups, namely, low BLL female (n = 47; BLL = 3.76 µg/dL), low BLL Male (n = 43; BLL = 4.13 µg/dL), high BLL female (n = 21; BLL = 23.5 µg/dL), and high BLL male (n = 18; BLL = 23.7 µg/dL), respectively. The low BLL group was associated with increased TNF-α levels, and the high BLL group was associated with reduced TNF-α levels in female subjects. No associations between BLL and the levels of IL-8 and TNF-α cytokines were observed in either females or males, respectively. A negative correlation between BLL and TNF-α was found in female subjects, suggesting that an increase in BLL accompanied by a reduction in TNF-α. The reduced levels of circulating TNF-α in female subjects suggest that chronic Pb exposure could predispose females to immune and inflammation-related disorders than their male counterparts. Further studies are recommended to ascertain the impact of chronic Pb exposure on immunomodulatory cytokines, especially in females.

An Evaluation of the Antibacterial Activity of Pterocarpus tinctorius Bark Extract against Enteric Bacteria That Cause Gastroenteritis.

Enteric bacteria are the leading cause of bacterial gastroenteritis worldwide, particularly in low-income countries. The bark decoction of Pterocarpus tinctorius (Fabaceae) has traditionally been used to treat bacterial gastroenteritis. However, studies reporting the antibacterial activity of Pterocarpus tinctorius are rare. Therefore, this study aimed to evaluate the antibacterial activity of stem bark extract of Pterocarpus tinctorius against Escherichia coli, Salmonella typhi, and Shigella dysenteriae. The powdered bark extract was successively extracted with methanol using the cold continuous maceration method, followed by partitioning the crude methanolic extract to obtain methanolic, hexane, and chloroform subextracts. Three fractions were isolated from the methanolic subextract using ordinary normal phase column chromatography. The antibacterial activity of the extracts and fractions was performed using the agar well diffusion method. The minimum inhibitory concentration (MIC) was determined using the agar well diffusion method. While, minimum bactericidal concentration (MBC) was obtained by the subculturing method. The methanolic subextract was the only extract that showed antibacterial activity against the tested bacteria, and its activity was highest on Shigella dysenteriae followed by Salmonella typhi and was least active on Escherichia coli, with mean inhibition zones of 14.3 ± 0.2, 13.7 ± 0.3, and 12.2 ± 0.1 at 200 mg/mL, respectively. Chloroform subextract showed antibacterial activity only on Shigella dysenteriae, while hexane subextract did not show antibacterial activity against all bacteria tested at 100 mg/mL and 200 mg/mL. Among the three subfractions of methanolic subextract, only one subfraction was active and had both mean minimum inhibitory concentration and a minimum bactericidal concentration against Escherichia coli at 1.25 mg/mL, Salmonella typhi at 1.25 mg/mL, and Shigella dysenteriae at 0.6 mg/mL. The findings of this study support the use of Pterocarpus tinctorius in traditional medicine. Therefore, purification and structural elucidation studies are highly recommended.

Methylation profiles of global LINE-1 DNA and the GSTP1 promoter region in children exposed to lead (Pb).

Lead (Pb) exposure has adverse health effects and altered DNA methylation may contribute to Pb toxicity. LINE-1 is an interspersed repeated DNA that is used as a surrogate marker for estimating genomic DNA methylation levels, and GSTP1 is an isozyme that detoxifies xenobiotics like Pb, and its expression is inhibited by methylation. Thus, to assess the effects of Pb exposure on global hypomethylation and gene-specific promoter hypermethylation, we examined DNA methylation at LINE-1 repetitive elements and the GSTP1 promoter region. Blood samples were obtained from children (N = 123) living in Pb-polluted areas (as exposed children) and children (N = 63) living in Pb-unpolluted areas (as control children) in Kabwe, Zambia. ICP-MS was used to determine blood lead levels (BLLs), and pyrosequencing and a fluorescence-based polymerase chain reaction assay were used to determine levels of LINE-1 methylation and GSTP1 promoter methylation, respectively. Inverse association was found between BLLs and LINE-1 methylation (ß = - 0.046, p = 0.006). The highest quartile of BLL had significant hypomethylation of LINE-1 (p for trend = 0.03), suggesting the higher the BLL, the lower LINE-1 methylation. GSTP1 methylation levels did not differ significantly between the two areas (p = 0.504), nor was it associated with Pb poisoning risk (OR = 1.03, p = 0.476), indicating GSTP1 methylation may not be a reliable biomarker of Pb exposure in healthy people. Therefore, Pb-related health problems could result from global DNA methylation changes due to high BLLs.

Narrative review of lead poisoning in humans caused by industrial activities and measures compatible with sustainable industrial activities in Republic of Zambia.

Lead (Pb) pollution and human exposure to Pb, is an important issue for the international community to address being associated with 0.90 million deaths from long-term effects. The Republic of Zambia is a typical mineral resource-rich country, with long-standing mining and smelting activities of metals including Pb in several parts of the country. This narrative review provides a comprehensive overview of previous papers that have assessed human exposure to Pb and related health effects in Zambia. Environmental remediation methods that should be applied locally, ways to reduce Pb exposure of the population, and issues that need to be addressed by various sectors are discussed. Environmental remediation methods using locally available and affordable materials are needed to ensure both sustainable industrial activities and pollution prevention. In the Zambian mining towns, including Kabwe, various research activities have been conducted, including environmental monitoring, human biomonitoring and health impact assessments. The town of Kabwe, which was one of Zambia's largest Pb mining area in the 20th century, continues to have formal and informal Pb-related industries and is known as one of the most polluted areas in the world. For example, despite the World Health Organization asserting that "For an individual with a blood Pb concentration ≥ 5 µg/dL, appropriate action should be taken to terminate exposure", there are reports of blood Pb levels in Kabwe children exceeding 100 µg/dL. While Pb pollution is a global issue, not many places have such continuous and comprehensive research has been conducted, and there is much to be learned from the knowledge accumulated in these areas. Because the high levels of Pb accumulation in humans and the adverse health effects were clarified, we consider that it is important to combine mining activities, which are a key industry, with measures to prevent environmental pollution.

Human Health Risk Assessment from Lead Exposure through Consumption of Raw Cow Milk from Free-Range Cattle Reared in the Vicinity of a Lead-Zinc Mine in Kabwe.

Lead (Pb) contamination in the environment affects both humans and animals. Chronic exposure to Pb via dietary intake of animal products such as milk from contaminated areas poses a health risk to consumers; therefore, the present study investigated Pb contamination in cow milk and its health risk impact on humans through consumption of milk from cattle reared in the proximity of a Pb−Zn mine in Kabwe, Zambia. Fresh milk samples were collected from cows from Kang'omba (KN), Kafulamse (KF), Mpima (MP), Mukobeko (MK), and Munga (MN) farming areas. Pb determination was performed using Graphite Flame Absorption Atomic Spectrophotometry (GFAAS). Cow milk Pb levels showed different concentration patterns according to season, distance, and location of the farms from the Pb−Zn mine. The overall mean Pb levels were ranged 0.60−2.22 µg/kg and 0.50−4.24 µg/kg in the wet and dry seasons, respectively. The mean Pb concentration, chronic daily intake (CDIs), target hazard quotients (THQs), and incremental lifetime cancer risk (ILCR) results obtained were all within the permissible limits of 20 µg/kg, 3 and 12.5 µg/kg-BW/day, <1 and 10−4 to10−6, respectively. In conclusion, although Pb was detected in milk from cows reared in Kabwe, the health risk effects of Pb exposure associated with the consumption of milk in both adults and children were negligible.

Metabolomic Alteration in the Plasma of Wild Rodents Environmentally Exposed to Lead: A Preliminary Study.

Lead poisoning is often considered a traditional disease; however, the specific mechanism of toxicity remains unclear. The study of Pb-induced alterations in cellular metabolic pathways is important to understand the biological response and disorders associated with environmental exposure to lead. Metabolomics studies have recently been paid considerable attention to understand in detail the biological response to lead exposure and the associated toxicity mechanisms. In the present study, wild rodents collected from an area contaminated with lead (N = 18) and a control area (N = 10) were investigated. This was the first ever experimental metabolomic study of wildlife exposed to lead in the field. While the levels of plasma phenylalanine and isoleucine were significantly higher in a lead-contaminated area versus the control area, hydroxybutyric acid was marginally significantly higher in the contaminated area, suggesting the possibility of enhancement of lipid metabolism. In the interregional least-absolute shrinkage and selection operator (lasso) regression model analysis, phenylalanine and isoleucine were identified as possible biomarkers, which is in agreement with the random forest model. In addition, in the random forest model, glutaric acid, glutamine, and hydroxybutyric acid were selected. In agreement with previous studies, enrichment analysis showed alterations in the urea cycle and ATP-binding cassette transporter pathways. Although regional rodent species bias was observed in this study, and the relatively small sample size should be taken into account, the present results are to some extent consistent with those of previous studies on humans and laboratory animals.

Assessment of heavy metals in wild and farmed tilapia (Oreochromis niloticus) on Lake Kariba, Zambia: implications for human and fish health.

The aim of this study was to assess the levels of heavy metals in both wild and farmed tilapia on Lake Kariba in Zambia and to evaluate the impact of intensive fish farming on wild tilapia. Three sites for wild fish (2 distant and 1 proximal to fish farms) and two fish farms were selected. One hundred fish (52 from distant sites; 20 near fish farms; 28 farmed fish) were sampled and muscle tissues excised for analysis of heavy metals (Mg, Fe, Zn, Al, Cu, Se, Co, Mo, As, Cr, V, Ni, Hg, Pb, Li, Cd, and Ag) by acid (HNO3) digestion and ICP-MS. All metals were found to be below the maximum limits (MLs) set by WHO/EU. Essential metals were higher in farmed tilapia, whereas non-essential metals were higher in wild tilapia. Significantly higher levels of essential metals were found in wild fish near the fish farms than those distant from the farms. Estimated weekly intake (EWI) for all metals were less than the provisional tolerable weekly intakes (PTWI). Target hazard quotients (THQ) and Hazard Indices (HI) were <1, indicating no health risks from a lifetime of fish consumption. Selenium Health Benefit Value (HBVSe) was positive for all locations, indicating protective effects of selenium against mercury in fish. Total cancer risk (CR) due to As, Cr, Cd, Ni and Pb was less than 1 × 10-4, indicating less than 1 in 10,000 carcinogenic risk from a lifetime consumption of tilapia from Lake Kariba. Hg levels (0.021 mg/kg) in wild tilapia at site 1 were higher than the Environmental quality standard (EQS = 0.020 mg/kg) set by EU, indicating possible risk of adverse effects to fish. Except for Hg, levels of metals in fish were safe for human consumption and had no adverse effects on fish.

Glutathione S-transferase gene polymorphisms in association with susceptibility to lead toxicity in lead- and cadmium-exposed children near an abandoned lead-zinc mining area in Kabwe, Zambia.

Interindividual genetic variations determine human's susceptibility to heavy metal-induced toxicity. Thus, we analyzed blood concentrations of lead (Pb) and cadmium (Cd) in 140 lead-exposed children. Genotyping of the glutathione S-transferase (GST) genes, GSTM1, GSTT1, and GSTP1 genes, was carried out to investigate their possible association with heavy metal concentrations and the risk of susceptibility to Pb toxicity. Exposure to both heavy metals was prevalent among the children. The blood Pb level ranged from 3.30 to 74.0 µg dL-1 with an average value of 26.8 µg dL-1 that is five times above its reference level. The average Cd level (0.22 µg L-1) was below its reference level. The metal-gene interaction showed positive correlation between GSTT1 null genotype and Pb and Cd levels (ß = 0.11; p = 0.02 and ß = 0.10; p = 0.01, respectively). More pronounced effects (ß = 0.19; p < 0.01 and ß = 0.25; p = 0.04) were found for the mixture of the three putative genes with blood Pb concentration. The susceptibility analysis using 10 µg dL-1 as blood Pb cutoff level showed a high risk of Pb toxicity (OR = 2.54; 95% CI: 1.02-6.32, p = 0.04) for children carrying the GSTP1 Ile/Val genotype. Further, the combined effect of GSTP1 Ile/Val with GSTT1 null genotype was more pronounced and showed an increased risk of susceptibility to Pb toxicity (OR = 11.7; 95% CI: 1.36-102.1, p = 0.02). In summary, this study suggests that GSTT1 null and GSTP1 Ile/Val genotypes are the main genetic factors, and individual and specific combinations of GSTP1 Ile/Val with GSTM1 and GSTT1 GST polymorphisms are associated with susceptibility to Pb toxicity.

Lead concentrations and isotope ratios in blood, breastmilk and feces: contribution of both lactation and soil/dust exposure to infants in a lead mining area, Kabwe, Zambia.

Lead (Pb) poses a serious public health concern. Breastmilk may be a possible source of Pb exposure in infants, as Pb can be transferred from the maternal blood to breastmilk. The present study was undertaken to determine the Pb exposure and the contribution of lactation as one of the exposure pathways to infants in a Pb mining area, Kabwe, Zambia. Blood, breastmilk and infants' feces were collected from 418 pairs of infants and mothers. The Pb concentrations, isotope ratios in the samples, and biochemistry in mothers' plasma were analyzed. The overall mean of blood lead levels (BLLs) in infants and mothers were 18.0 and 11.3 µg/dL, respectively. High Pb concentration in breastmilk (range: 0.4-51.9, mean: 5.3 µg/L) above the WHO acceptable level between 2 and 5 µg/L were found and could be one of the sources of Pb exposure in infants. The Pb isotope ratios in infants' feces were the most similar to Pb ratios in the soil samples. The results suggest that infants are also exposed to Pb from the environment. Pb exposure in infants through breastfeeding and soil ingestion could potentially exceed daily intake of Pb which causes neurodevelopmental toxicity. In contrast to the high BLLs in mothers, the plasma biochemical profiles of most analyzed parameters were interestingly within, or close to, the standard reference values. Our data suggest that environmental remediation is urgently needed to reduce the Pb exposure in infants and mothers from the environment in Kabwe in parallel with chelation therapy.

Genome-wide DNA methylation analysis of dogs with high lead exposure living near a lead mining area in Kabwe, Zambia.

Lead (Pb) is a heavy metal that has been proven to be toxic to both animals and humans. Genom-wide DNA methylation in domestic dogs exposed to high levels of Pb in Kabwe, Zambia was analyzed in this study. Using next-generation sequencing on samples from 20 domestic dogs (mean blood Pb concentration: 43.6 µg/dL and 7.2 µg/dL in the high and low exposure groups), a digital restriction enzyme analysis of methylation was performed to identify the genomic locations of differentially methylated CpG sites. A validation study on an additional 20 dogs followed (blood Pb concentration: 4.9-29.7 µg/dL). The cluster analysis resolved two broad clusters indicating high and low Pb exposure. The study identified 827 (1.2%) CpG sites with differences in methylation (101 CpG sites were hypermethylated in the low exposure group and 726 were hypermethylated in the high exposure group). The sites corresponded to 26 genes with differentially methylated CpG sites at their promoter regions, including the NGF gene. The methylation of four CpG sites was validated using bisulfite pyrosequencing. The results indicate that aberrant hypermethylation is prevalent in dogs exposed to Pb. The altered DNA methylation of the genes identified in this study contributes to a greater understanding of the epigenetic changes caused by Pb exposure and highlights novel biomarker discoveries across species.

Assessment of LeadCare® II analysis for testing of a wide range of blood lead levels in comparison with ICP-MS analysis.

The LeadCare® testing system, which utilizes anodic stripping voltammetry (ASV) methodology, has been widely used worldwide for cost-effective blood lead level (BLL) screening. However, some concerns have recently been issued regarding inaccurate results obtained using LeadCare®. Hence, we aimed to evaluate the accuracy of BLL measured by LeadCare® II (BLLLC) by comparison with ICP-MS (BLLIM) by the Passing-Bablok regression, Deming regression, and Bland-Altman analyses by using 994 venous blood samples. BLLLC ranged from 3.3 to 162.3 µg/dL, while BLLIM ranged from 0.8 to 154.8 µg/dL. Although BLLLC and BLLIM exhibited a strong and positive correlation, BLLLC values were generally greater than BLLIM values, indicative of the overestimation of the LeadCare® analysis. A large positive bias of 19.15 ± 8.26 µg/dL and 29.25 ± 14.04 µg/dL for BLLLC compared with BLLIM were recorded in the BLLLC range of 45.0-64.9 µg/dL and for ≥65.0 µg/dL, respectively. In contrast, a bias of ≤0.3 µg/dL was observed at a BLLLC of less than 10.0 µg/dL. Blood copper, cadmium, and iron levels did not exhibit an effect on the bias of BLLLC, indicative of the minimal potential interferences of the metals; these interferences are a cause for concern with the ASV method. In conclusion, LeadCare® analysis is thought to be a good tool for screening purposes at a lower BLL around the reference level of 5 µg/dL in the initial stage; however, conversion or retesting using a laboratory analyzer is recommended at a higher BLL for appropriate clinical evaluation and research.

An Investigation of the Wild Rat Crown Incisor as an Indicator of Lead (Pb) Exposure Using Inductively Couple Plasma Mass Spectrometry (ICP-MS) and Laser Ablation ICP-MS.

Lead (Pb) is a metal toxicant of great public health concern. The present study investigated the applicability of the rat incisor in Pb exposure screening. The levels of lead in teeth (Pb-T) in the crown and root of incisors in laboratory Pb-exposed Sprague Dawley rats were quantified using inductively coupled plasma mass spectrometry (ICP-MS). The crown accumulated much Pb-T than the root of the Sprague Dawley rat incisor. The levels of lead in blood (Pb-B) were positively correlated with the Pb-T in the crown and root incisors of the Sprague Dawley rats. As an application of the Pb-T crown results in experimental rats, we subsequently analyzed the Pb-T in the crown incisors of Pb-exposed wild rats (Rattus rattus) sampled from residential sites within varying distances from an abandoned lead-zinc mine. The Pb-T accumulation in the crown of incisors of R. rattus rats decreased with increased distance away from the Pb-Zn mine. Furthermore, the Pb-T was strongly correlated (r = 0.85) with the Pb levels in the blood. Laser ablation ICP-MS Pb-T mappings revealed a homogenous distribution of Pb in the incisor with an increased intensity of Pb-T localized in the tip of the incisor crown bearing an enamel surface in both Sprague Dawley and R. rattus rats. These findings suggest that Pb-T in the crown incisor may be reflective of the rat's environmental habitat, thus a possible indicator of Pb exposure.

Clinical biochemical parameters associated with the exposure to multiple environmental metals in residents from Kabwe, Zambia.

Lead (Pb) interferes with various bodily functions. Although high blood Pb (Pb-B) levels in residents from Kabwe, Zambia have been reported, the accumulation pattern of other metals remains unknown. The study was designed to determine the Pb-B, blood cadmium (Cd-B), and zinc (Zn-B) values of 504 representative samples from Kabwe, as well as the potential associated adverse health effects. The Pb-B level ranged from 0.79 to 154.75 µg/dL and generally increased in areas near the mine. A significant elevation of Cd-B was observed in two areas (0.37 ± 0.26 and 0.32 ± 0.30 µg/L) where the two highest mean Pb-B levels were recorded. By contrast, the Zn-B values did not differ greatly with respect to area. Some blood biochemical parameters relating to hepatic and renal functions were out of the normal range in approximately 20-50% of studied adult participants. The δ-aminolevulinic acid dehydratase (δ-ALAD) activity was significantly inhibited in the two areas contaminated by Pb and Cd. A significant negative relationship was observed between metal levels and clinical parameters, e.g., between Pb-B and δ-ALAD for all the age categories and between Cd-B and the estimated glomerular filtration rate for all the age categories except 0-4 years. The elevated Cd-B in areas near the mine relative to the other areas suggested the potential adverse health effects of Cd and/or the interaction of Pb and Cd. A significant association of metal levels with clinical parameters also indicated the effects of metal exposure on hematopoietic, hepatic, and renal systems.

Land Use in Habitats Affects Metal Concentrations in Wild Lizards Around a Former Lead Mining Site.

We investigated the potential effects of different land use and other environmental factors on animals living in a contaminated environment. The study site in Kabwe, Zambia, is currently undergoing urban expansion, while lead contamination from former mining activities is still prevalent. We focused on a habitat generalist lizards (Trachylepis wahlbergii). The livers, lungs, blood, and stomach contents of 224 lizards were analyzed for their lead, zinc, cadmium, copper, nickel, and arsenic concentrations. Habitat types were categorized based on vegetation data obtained from satellite images. Multiple regression analysis revealed that land use categories of habitats and three other factors significantly affected lead concentrations in the lizards. Further investigation suggested that the lead concentrations in lizards living in bare fields were higher than expected based on the distance from the contaminant source, while those in lizards living in green fields were lower than expected. In addition, the lead concentration of lungs was higher than that of the liver in 19% of the lizards, implying direct exposure to lead via dust inhalation besides digestive exposure. Since vegetation reduces the production of dust from surface soil, it is plausible that dust from the mine is one of the contamination sources and that vegetation can reduce exposure to this.

Blood lead levels and aberrant DNA methylation of the ALAD and p16 gene promoters in children exposed to environmental-lead.

BACKGROUND: Lead (Pb) is a well-known toxic heavy metal which can have serious public health hazards. As of today, there is no safe threshold for Pb exposure, especially for children. Lead exposure has been associated with adverse health outcomes involving epigenetic mechanisms, such as aberrant DNA methylation. The objective of the present study was to elucidate the associations between blood lead levels (BLLs) and gene-specific promoter DNA methylation status in environmental Pb-exposed children from Kabwe, Zambia. METHODS: A cross-sectional study was conducted using 2 to 10-year-old children from high Pb exposed area (N = 102) and low Pb exposed area (N = 38). We measured BLLs using a LeadCare II analyzer and investigated the methylation status of the ALAD and p16 gene promoters by methylation-specific PCR. RESULTS: The mean BLLs were 23.7 µg/dL and 7.9 µg/dL in high Pb exposed and low Pb exposed children, respectively. Pb exposure was correlated with increased methylation of the ALAD and p16 genes. The promoter methylation rates of ALAD and p16 in high Pb exposed children were 84.3% and 67.7%, and 42.1% and 44.7% in low Pb exposed children, respectively. Significantly increased methylation was found in both genes in high Pb exposed children compared with low Pb exposed children (p < 0.05). Children with methylated ALAD and p16 genes showed an increased risk of Pb poisoning (odd ratio >1) compared to the unmethylated status. CONCLUSIONS: This study for the first time tries to correlate promoter methylation status of the ALAD and p16 genes in environmental Pb-exposed children from Kabwe, Zambia as a representative. The result suggests that Pb exposure increases aberrations in ALAD and p16 gene methylation, which may be involved in the mechanism of Pb toxicity.

Factors associated with lead (Pb) exposure on dogs around a Pb mining area, Kabwe, Zambia.

Lead (Pb)-poisoning is a serious public health concern and dogs have been useful as a sentinel-animal for Pb exposure of humans. In the present study, the blood Pb concentrations (BLC), isotope ratios (208 Pb/206 Pb and 207 Pb/206 Pb), and biochemistry of 120 domestically owned dogs living around a Pb mining area, in Kabwe, Zambia were analyzed to determine factors associated with Pb exposure. The overall mean value of Pb in dog blood in the present study was 271.6 µg/L. The BLC in the dogs from sites near the mine were significantly higher than those in the dogs from a site 4 km from the mine (352.9 ± 205.1 µg/L versus 28.0 ± 13.9 µg/L). BLC significantly decreased with both increasing age of the dogs and distance from the mine. The Pb isotope ratios in the dog that resided near the mine showed values similar to those reported at the galena mine in Kabwe, which is considered to be the source of Pb exposure. In contrast to the high metal exposure that was determined in these dogs, the mean values of most analyzed parameters in the blood biochemical analysis were surprisingly within or close to the standard reference values. Moreover, none of the dogs showed overt signs of Pb-poisoning or other clinical symptoms. The results of analysis of Pb exposure of the dogs obtained in the present study, which are similar to the previously reported results in human in this location, suggest that dogs could be useful as a sentinel animal for Pb exposure of humans in Kabwe.

Current trends of blood lead levels, distribution patterns and exposure variations among household members in Kabwe, Zambia.

Childhood lead (Pb) poisoning has devastating effects on neurodevelopment and causes overt clinical signs including convulsions and coma. Health effects including hypertension and various reproductive problems have been reported in adults. Historical Pb mining in Zambia's Kabwe town left a legacy of environmental pollution and childhood Pb poisoning. The current study aimed at establishing the extent of Pb poisoning and exposure differences among family members in Kabwe as well as determining populations at risk and identify children eligible for chelation therapy. Blood samples were collected in July and August 2017 from 1190 household members and Pb was measured using a portable LeadCare-II analyser. Participants included 291 younger children (3-months to 3-years-old), 271 older children (4-9-years-old), 412 mothers and 216 fathers from 13 townships with diverse levels of Pb contamination. The Blood Lead Levels (BLL) ranged from 1.65 to 162  µg/dL, with residents from Kasanda (mean 45.7  µg/dL) recording the highest BLL while Hamududu residents recorded the lowest (mean 3.3  µg/dL). Of the total number of children sampled (n = 562), 23% exceeded the 45  µg/dL, the threshold required for chelation therapy. A few children (5) exceeded the 100  µg/dL whereas none of the parents exceeded the 100  µg/dL value. Children had higher BLL than parents, with peak BLL-recorded at the age of 2-years-old. Lead exposure differences in Kabwe were attributed to distance and direction from the mine, with younger children at highest risk. Exposure levels in parents were equally alarming. For prompt diagnosis and treatment, a portable point-of-care devise such as a LeadCare-II would be preferable in Kabwe.

One year exposure to Cd- and Pb-contaminated soil causes metal accumulation and alteration of global DNA methylation in rats.

Metal pollution has been associated with anthropogenic activities, such as effluents and emissions from mines. Soil could be exposure route of wild rats to metals, especially in mining areas. The aim of this study was to verify whether soil exposure under environmentally relevant circumstances results in metal accumulation and epigenetic modifications. Wistar rats were divided to three groups: 1) control without soil exposure, 2) low-metal exposure group exposed to soil containing low metal levels (Pb: 75 mg/kg; Cd: 0.4), and 3) high-metal exposure group exposed to soil (Pb: 3750; Cd: 6). After 1 year of exposure, the metal levels, Pb isotopic values, and molecular indicators were measured. Rats in the high-group showed significantly greater concentrations of Pb and Cd in tissues. Higher accumulation factors (tissue/soil) of Cd than Pb were observed in the liver, kidney, brain, and lung, while the factor of Pb was higher in the tibia. The obtained results of metal accumulation ratios (lung/liver) and stable Pb isotope ratios in the tissues indicated that the respiratory exposure would account for an important share of metal absorption into the body. Genome-wide methylation status and DNA methyltransferase (Dnmt 3a/3b) mRNA expressions in testis were higher in the high-group, suggesting that exposure to soil caused metal accumulation and epigenetic alterations in rats.