Adekunle A. Bakare

Expression of the cytochrome P450s, CYP6P3 and CYP6M2 are significantly elevated in multiple pyrethroid resistant populations of Anopheles gambiae s.s . from Southern Benin and Nigeria

BackgroundInsecticide resistance in Anopheles mosquitoes is threatening the success of malaria control programmes. This is particularly true in Benin where pyrethroid resistance has been linked to the failure of insecticide treated bed nets. The role of mutations in the insecticide target sites in conferring resistance has been clearly established. In this study, the contribution of other potential resistance mechanisms was investigated in Anopheles gambiae s.s. from a number of localities in Southern Benin and Nigeria. The mosquitoes were sampled from a variety of breeding sites in a preliminary attempt to investigate the role of contamination of mosquito breeding sites in selecting for resistance in adult mosquitoes.ResultsAll mosquitoes sampled belonged to the M form of An. gambiae s.s. There were high levels of permethrin resistance in an agricultural area (Akron) and an urban area (Gbedjromede), low levels of resistance in mosquito samples from an oil contaminated site (Ojoo) and complete susceptibility in the rural Orogun location. The target site mutation kdrW was detected at high levels in two of the populations (Akron f = 0.86 and Gbedjromede f = 0.84) but was not detected in Ojoo or Orogun. Microarray analysis using the Anopheles gambiae detox chip identified two P450s, CYP6P3 and CYP6M2 up regulated in all three populations, the former was expressed at particularly high levels in the Akron (12.4-fold) and Ojoo (7.4-fold) populations compared to the susceptible population. Additional detoxification and redox genes were also over expressed in one or more populations including two cuticular pre-cursor genes which were elevated in two of the three resistant populations.ConclusionMultiple resistance mechanisms incurred in the different breeding sites contribute to resistance to permethrin in Benin. The cytochrome P450 genes, CYP6P3 and CYP6M2 are upregulated in all three resistant populations analysed. Several additional potential resistance mechanisms were also identified that warrant further investigation. Metabolic genes were over expressed irrespective of the presence of kdr, the latter resistance mechanism being absent in one resistant population. The discovery that mosquitoes collected from different types of breeding sites display differing profiles of metabolic genes at the adult stage may reflect the influence of a range of xenobiotics on selecting for resistance in mosquitoes.

Comparative evaluation of environmental contamination and DNA damage induced by electronic-waste in Nigeria and China.

In the last decade, China and Nigeria have been prime destinations for the worlds e-waste disposal leading to serious environmental contamination. We carried out a comparative study of the level of contamination using soils and plants from e-waste dumping and processing sites in both countries. Levels of polyaromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and polybrominated diphenyl ethers (PBDEs) were analyzed using gas chromatography/spectrophotometry and heavy metals using atomic absorption spectrophotometry. DNA damage was assayed in human peripheral blood lymphocytes using an alkaline comet assay. Soils and plants were highly contaminated with toxic PAHs, PCBs, PBDEs, and heavy metals in both countries. Soil samples from China and plant samples from Nigeria were more contaminated. There was a positive correlation between the concentrations of organics and heavy metals in plant samples and the surrounding soils. In human lymphocytes, all tested samples induced significant (p<0.05) concentration-dependent increases in DNA damage compared with the negative control. These findings suggest that e-waste components/constituents can accumulate, in soil and surrounding vegetation, to toxic and genotoxic levels that could induce adverse health effects in exposed individuals.

Does the spillage of petroleum products in Anopheles breeding sites have an impact on the pyrethroid resistance

BackgroundThe emergence of Anopheles populations capable of withstanding lethal doses of insecticides has weakened the efficacy of most insecticide based strategies of vector control and, has highlighted the need for further studies on the mechanisms of insecticide resistance and the various factors selecting resistant populations of mosquitoes. This research targeted the analysis of breeding sites and the oviposition behaviour of susceptible and resistant populations of Anopheles in localities of spilled petroleum products. The aim was to establish the possible contribution of oil spillage in the selection of pyrethroid resistance in malaria vectors.MethodsAnopheles breeding sites were identified and the insecticide susceptibility of the Anopheles gambiae populations mapped in 15 localities of South Western Nigeria. The presence of oil particles as well as the turbidity, the dissolved oxygen and the pH of each identified breeding site was recorded. Data were cross-analysed to correlate the habitat types and the insecticide susceptibility status of emerging mosquitoes. The second phase of this study was basically a laboratory model to provide more information on the implication of the spillage of petroleum on the selection of pyrethroid resistance in An. gambiae.ResultsModerate levels of resistance following exposure to permethrin-impregnated papers were recorded with the majority of An. gambiae samples collected in the South Western Nigeria. Data from this study established a link between the constituency of the breeding sites and the resistance status of the emerging Anopheles.ConclusionThis study has revealed the segregational occupation of breeding habitats by pyrethroid resistant and susceptible strains of An. gambiae in south-western Nigeria. Compiled results from field and laboratory research point out clear relationships between oil spillage and pyrethroid resistance in malaria vectors. The identification of this factor of resistance could serve as strong information in the management of insecticide resistance in some West African settings.

Cytogenotoxic effects of electronic waste leachate in Allium cepa

Information on the potentials of electronic waste (e-waste) leachate to induce genotoxicity in eukaryotic cells is scarce. In this study, we investigated the potential of raw and simulated e-waste leachates to induce cytotoxicity and DNA damage in Allium cepa. Roots of onion bulbs were grown in five concentrations: 1, 5, 10, 25 and 50% (v/v; leachate/tap water) of each of the leachate sample, tap water (negative control) and 0.25 ppm lead nitrate (positive control). Cytogenetic and root length inhibition analyses were evaluated at 48 and 72 hours respectively. There was concentration-dependent significant (p < 0.05) inhibition of root growth and cell proliferation compared to the negative control. The leachates also induced morphological modifications of the roots and cytological aberrations such as anaphase bridge, sticky chromosomes and binucleate cells. Fe, Cd, Mn, Cu, Ni, Cr As, Zn and Pb analysed in the samples were believed to play significant role in the observed cytogenotoxicity in A. cepa. These observations indicate that e-waste leachate contained substances capable of inducing cytotoxicity and somatic mutations in A. cepa. This is of environmental and public health importance in Nigeria and other developing nations whose populations are at risk of e-waste exposure.

Genotoxicity and mutagenicity of electronic waste leachates using animal bioassays

Human exposure to electronic waste (e-waste) is increasing, especially in developing nations. This has potential short and chronic adverse health risks. Information is currently scarce on potential genotoxicity of e-waste leachate. In this study, the potential mutagenic and genotoxic effects of leachates from e-waste dumpsites at two electronic markets in Lagos, Nigeria, were investigated. The assays utilized were: bone marrow micronucleus (MN) and chromosome aberration (CA), spermatogonia CA, sperm morphology, and sperm count in mice. Experiments were carried out at concentrations of 1%, 5%, 10%, 25%, and 50% (v/v; leachate: distilled water) of leachate samples. MN analysis showed a concentration-dependent induction of micronucleated polychromatic erythrocytes (MNPCE) across the treatment groups. In the CA tests (bone marrow and spermatogonia), there was concentration-dependent significant reduction in mitotic index and induction of different types of CA. Assessment of sperm shape showed a significant increase in sperm abnormalities with significant decrease in mean sperm count in treated groups. Heavy metals analyzed in the tested samples are believed to contribute significantly to the observed genetic damage. This indicates that e-waste contains potential genotoxic agents; and constitutes a genetic risk in exposed human population.

Frequency of twinning in southwest Nigeria

BACKGROUND: In the human species, twin is a type of multiple birth in which the mother gives birth to two offspring from the same pregnancy. The occurrence and frequency of twinning, however, varies across human populations. The maternal age, socio-environmental factors, increase in the use of contraceptives, the race of human population, increase in the spontaneous abortion rate, and seasonal variations are among the factors that could influence twinning rate. Information on twinning rates in southwest Nigeria is limited. AIMS: This study presents information on the frequency of twinning, as well as its analysis by maternal age, in four urban settings in southwest Nigeria. This is with the aim of extending current knowledge on the frequency of twinning in southwest Nigeria and contributing to the demographic studies in the country. MATERIALS AND METHODS: Data on single births and twin births from January 1995 to December 2004 were collected from the Oyo State General Hospital (OSGH), Wesley Guild Hospital (WGH), Obafemi Awolowo University Teaching Hospital (OAUTH), and Ekiti State Specialist Hospital (ESSH) in Ogbomoso, Ilesa, Ile-Ife, and Ado-Ekiti respectively. These were analyzed by year and maternal age groups of 15-19, 20-24, 25-29, 30-34, 35-39, 40-44, and 45-49 years according to the standard method. RESULTS: A frequency of twin births of 46.5 per 1000 deliveries and 46.2 per 1000 deliveries was recorded for Ilesa and Ile-Ife respectively. The frequency recorded for Ogbomoso and Ado-Ekiti was 38.5 and 22.1 per 1000 deliveries respectively. The overall average frequency of 40.2 per 1000 deliveries for the four hospitals ranks among the highest recorded rates of twin births in the world. The maternal age group of 25-29 years had the highest occurrence of twin births, while the lowest was recorded in the 45-49 years age group. CONCLUSION: This analysis reveals high incidence of twinning in the studied areas and supports previous assertion that the southwestern part of Nigeria has the highest twinning rate in the country and in the whole world. It is our opinion that diet, maternal history of twinning, and some socio-environmental factors may have influenced the results.

Genotoxicity and cytotoxicity of chromium, copper, manganese and lead, and their mixture in WIL2-NS human B lymphoblastoid cells is enhanced by folate depletion

Heavy metal exposure or dietary deficiency is associated with increased genetic damage, cancer and age-related diseases. Folate (vitamin B9) required for DNA repair and synthesis may increase cellular susceptibility to metal induced genotoxicity. This study investigated the interactive effects of folic acid deficiency and sufficiency on genome instability and cytotoxicity induced by chromium (VI), copper (II), manganese (II), lead (IV), and their mixture (CCMP) in WIL2-NS human B lymphoblastoid cells. WIL2-NS cells were cultured in folic acid deficient (20 nM) and replete (2000 nM) RPMI 1640 medium treated with different concentrations (0.00-1000 μM) of the metals and CCMP for 48 h. Chromosomal damage and cytotoxicity were measured using the Cytokinesis-block Micronucleus Cytome assay. CCMP, Cr, Pb, Cu and Mn induced concentration dependent, increases in cells with chromosome damage (micronuclei, nucleoplasmic bridges, nuclear buds) and necrotic cells and decreased nuclear division index. The metals exhibited different cytotoxic and genotoxic potentials (CCMP>Cr>Pb>Cu>Mn) in both folate deficient and sufficient cells, with the cytogenotoxic effects being greater in folate deficient cells. Significant interaction between the metals and folic acid suggests that folic acid deficiency exacerbated cell proliferation inhibition and genome instability induced by metals. Folate deficiency, increasing metal concentration, and their interactions explained 3-11%, 74-92% and 4-12% of the variance of DNA damage biomarkers. In conclusion, exposure to the tested metals (0.01-1000 μM) increased chromosomal DNA damage in WIL2-NS cells and this was exacerbated by folate deficiency.

Genotoxic and anti-genotoxic effect of aqueous extracts of Spondias mombin L., Nymphea lotus L. and Luffa cylindrica L. on Allium cepa root tip cells

Spondias mombin L., Nymphea lotus L. and Luffa cylindrica L. are commonly used in the traditional management of cancer in Nigeria. The genotoxic and anti-genotoxic effects of aqueous extracts of these plants was evaluated using the Allium cepa L. assay. Onion bulbs were exposed to 0.5–10 mg ml–1 concentrations (v/v) of each extract, prepared as practiced locally, for analyses of root growth inhibition and induction of cytogenetic damage, respectively. There was concentration-dependent, significant (p < 0.05) inhibition of root growth by the extracts when compared with the negative control. The EC50 for the decoctions of S. mombin, N. lotus and L. cylindrica were 1.3, 1.8 and 16.2 mg ml–1 respectively. All the extracts had mitodepressive effects on cell division, and induced chromosomal aberrations (p ˂ 0.05). They showed chemopreventive activity through reduction of cytological aberration induced by lead nitrate. These findings suggest inhibitory, anti-proliferative and anti-cytogenetic damage activities of aqueous extracts of the tested medicinal plants on A. cepa.

DNA and oxidative damage induced in somatic organs and tissues of mouse by municipal sludge leachate

Pollution by waste landfill leachate has prompted a number of studies on the toxic and potential health effects. This study assessed the genotoxicity of a municipal sludge leachate (MSL) in the somatic tissues (blood and bone marrow) and organs (liver, kidney, and spleen) of mice using the alkaline Comet assay. The possible cause of DNA damage via the study of antioxidant system (lipid peroxidation [LPO]; catalase [CAT]; reduced glutathione [GSH]; and superoxide dismutase [SOD]) responses in mouse liver was also investigated. Different concentrations (2.5%, 5%, 10%, and 15%) of the leachate were administered intraperitoneally for 5 consecutive days to male Swiss albino mice (4 mice/group). A significant (p < 0.05) increase in DNA damage in organs and tissues of treated mice compared to the negative control was observed as evident from the Comet assay parameters: olive tail moment (OTM, arbitrary units) and tail DNA (%). Bone marrow showed maximum DNA damage followed by liver > spleen > kidney > blood as evident by the OTM. A significant increase (p < 0.05) in the level of antioxidant enzymes (CAT and SOD) and LPO with a concurrent decrease in GSH in the liver of treated mice was also observed. Our finding demonstrates that the MSL induces DNA damage in the somatic tissues and organs of mouse as well as induces oxidative stress in the liver. These tissues and organs may be the potential targets in animal and human populations exposed to MSL. This is of relevance to public health; as such exposure could lead to adverse health effects via systemic genotoxicity.

Quantification of the efficiency of treatment of Anopheles gambiae breeding sites with petroleum products by local communities in areas of insecticide resistance in the Republic of Benin

BackgroundThe emergence of Anopheles populations capable of withstanding lethal doses of insecticides has weakened the efficacy of most insecticide based strategies of vector control and, has highlighted the need for developing new insecticidal molecules or, improving the efficacy of existing insecticides or abandoning those to which resistance has emerged. The use of petroleum products (PP) against mosquito larvae had an immense success during early programmes of malaria control, but these compounds were abandoned and replaced in the 1950s by synthetic insecticides probably because of the high performances given by these new products. In the current context of vector resistance, it is important to elucidate the empirical use of PP by quantifying their efficiencies on resistant strains of Anopheles.MethodsLarvae of Anopheles Ladji a local resistant strain were exposed to increasing concentrations of various PP (kerosene, petrol and engine oils) for 24 hours and the lethal activities recorded. The highest concentration (HiC) having no lethal activity (also referred as the NOEL or no effect level) and the lowest concentration (LoC100) yielding 100% mortality were rated for each PP on the Ladji strain. Prior to laboratory analysis, KAP studies were conducted in three traditional communities were insecticide resistance is clearly established to confirm the use of PP against mosquitoes.ResultsLaboratory analysis of petrol, kerosene and engine oils, clearly established their lethal activities on resistant strains of Anopheles larvae. Contrary to existing references, this research revealed that exposed larvae of Anopheles were mostly killed by direct contact toxicity and not by suffocation as indicated in some earlier reports.ConclusionThis research could serve as scientific basis to backup the empirical utilisation of PP on mosquito larvae and to envisage possibilities of using PP in some traditional settings where Anopheles have developed resistance to currently used insecticides.