Landfill soil leachates from Nigeria and India induced DNA damage and alterations in genes associated with apoptosis in Jurkat cell.

Landfill soil leachates, containing myriad of xenobiotics, increase genotoxic and cytotoxic stress-induced cell death. However, the underlying mechanism involved in the elimination of the damaged cells is yet to be fully elucidated. This study investigated the apoptotic processes induced in lymphoma (Jurkat) cells by landfill soil leachates from Olusosun (OSL, Nigeria) and Nagpur (NPL, India). Jurkat was incubated with sub-lethal concentrations of OSL and NPL for 24 h and analyzed for DNA fragmentation and apoptosis using agarose gel electrophoresis and Hoechst 33258-PI staining, respectively. Complementary DNA expression profiling of some pro-apoptotic and anti-apoptotic genes regulating apoptosis was also analyzed using real-time PCR (RT-PCR) method. Agarose gel electrophoresis revealed DNA fragmentations in OSL and NPL-treated cells. Hoecsht-33258 - Propidium Iodide (PI) based apoptotic analysis confirmed apoptotic cell death in exposed Jurkat. RT-PCR analysis revealed different fold changes in the pro- and anti-apoptotic genes in OSL and NPL-treated Jurkat. There was significant increase in fold change of the up-regulated genes; apoptosis inducing factor mitochondrion-associated 2 (AIFM2), Fas-associated death domain (FADD), Caspase-2, Caspase-6, BH3 interacting domain death agonist (BID), tumor suppressor (p53), and BCL2 associated agonist of cell death (BAD) and down-regulation of apoptosis inhibitor 5 (API5). Results suggest that OSL and NPL elicited genotoxic stress-related apoptosis in Jurkat. The dysregulation in the expression of genes involved in apoptotic processes in wildlife and human exposed to landfill emissions may increase aetiology of various pathological diseases including cancer.

Metal Bioaccumulation, Cytogenetic and Clinico-Biochemical Alterations in Rattus norvegicus Exposed In Situ to a Municipal Solid Waste Landfill in Lagos, Nigeria.

This study aimed at determining in animal model the health effects of in situ exposure to landfill chemicals. We evaluated metal concentrations in tissues and cytogenetic and clinico-biochemical effects in Wistar rats (Rattus norvegicus) exposed in situ at Olusosun landfill in Lagos, Nigeria. Male rats (n = 30/point) were exposed at three different points to ambient air and underground water (via drinking) at the landfill for 4-, 8-, 12-, 16-, 20- and 24-week periods. Rats concurrently sited at a residential area, 17.3 km from the landfill site served as control. There was significantly (p < 0.05) time-dependent: accumulation of lead, cadmium, chromium, copper and zinc in the liver and kidney and increase in body weight gain, in exposed rats compared to control. There was significant induction of micronuclei and cytotoxicity (reduced PCE/NCE ratios) in exposed rats. Haematological parameters (RBC, PCV, Hb and WBC) and serum biomarkers of hepato-renal damage [aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH) activities; creatinine and urea levels] revealed significant increases. There was significant increase in hepatic levels of reduced glutathione, malondialdehyde, catalase activities, and decrease in superoxide dismutase, at all periods. Chromium and copper concentrations in the liver and kidney revealed significant positive correlations with either one or more of AST, ALT, LDH and urea. Significant metal concentrations in the underground water and tissues suggest that heavy metals are responsible for the observed alterations, and this may have been via oxidative stress. These findings suggest potential health risk due to occupational and residential exposure to landfill pollutants.

Wild black rats (Rattus rattus Linnaeus, 1758) as zoomonitor of genotoxicity and systemic toxicity induced by hazardous emissions from Abule Egba unsanitary landfill, Lagos, Nigeria.

Wild black rats (Rattus rattus) inhabiting Abule Egba landfill (AEL) were used as zoomonitor to assess health risk associated with exposure to hazardous chemicals from landfills. Twenty five R. rattus (16♂ and 9♀) captured within AEL and 15 (9♂ and 6♀) (control) caught from Iyano Ipaja (10 km away from AEL) were examined for bone marrow micronucleated polychromatic erythrocytes (MNPCE) and polychromatic erythrocytes/normochromatic erythrocytes (PCE/NCE) ratio, abnormal sperm morphology, alterations in hematological indices and erythrocyte morphology, and histopathology of the viscera using standard protocols. There was significant (p < 0.05) increase in MNPCE but decrease PCE/NCE ratio in bone marrow cells of exposed rats than the reference site. MNPCE was insignificantly higher in male than females. Cauda epididymal sperms from exposed rats showed significant high frequency of teratozoospermia. Erythrocyte count, hemoglobin concentrations, percentage hematocrits, mean corpuscular hemoglobin concentrations, leucocyte count, and lymphocytes decreased while mean corpuscular volume, neutrophils, and mean corpuscular hemoglobin increased in the exposed rats compared to the control. Also, abnormal erythrocyte morphology: acanthocytes, codocytes (target cells), schizocytes, and tear drops significantly increased in the exposed rats. Marginal sexual dimorphism was observed between males and females in the incidence of hematological indices. Histopathological lesions including interstitial edema, hemorrhage, lymphoid depletion, cellular infiltrations, proliferation of the alveolar pneumocytes, necrosis, tissue degeneration, and reduced germinal epithelium were observed in the testes, liver, lungs, heart, kidney, and spleen from the exposed rats compared to the control. Some physicochemicals and metals analyzed in leachates from the landfill are capable of inducing genome instability and systemic toxicity in the exposed rats. Rattus rattus exposed to hazardous chemicals from AEL harbored somatic and germ cell mutations, and tissue damage compared to the control rats. We suggest that R. rattus are useful sentinel for genotoxicity and system toxicity assessment of landfill-polluted sites.Graphical abstract.

Titanium dioxide nanoparticles-induced cytogenotoxicity and alterations in haematological indices of Clarias gariepinus (Burchell, 1822).

The application of titanium dioxide (TiO2) nanoparticles (NPs) in the manufacturing of consumer products has increased tremendously and with the potential to induce deleterious effects on aquatic biota. There have been reports on metal oxide NP toxicity in aquatic organisms, however, information on cytotoxicity and genotoxicity of TiO2 NPs on the African catfish, Clarias gariepinus, is scarce. In this study, we investigated the genotoxicity and haematotoxicity of TiO2 NPs in C. gariepinus using the micronucleus (MN) assay and haematological analysis, respectively. Juvenile C. gariepinus were exposed to 6.25, 12.5, 25.0, 50.0 and 100.0 mg L-1 concentrations of TiO2 NPs for 7 and 28 days. Benzene (0.05 mL L-1) and dechlorinated tap water were used as positive and negative controls, respectively. Data of the MN showed a significant (p < 0.05) concentration-dependent increase in the frequency of MN at both exposure periods in comparison to negative control. Red blood cells, haematocrit, platelets and heterophils significantly reduced with an increased mean corpuscular haemoglobin concentration and lymphocytes at the 7-day exposure period, while in the 28-day exposure period, mean cell volume, mean corpuscular haemoglobin and lymphocytes had a significant increase in comparison with the negative control. This study indicates that TiO2 NPs induced cytogenetic and haematological alterations in C. gariepinus and is of relevance in biodiversity and aquatic health management.

Alteration of sperm parameters and reproductive hormones in Swiss mice via oxidative stress after co-exposure to titanium dioxide and zinc oxide nanoparticles.

In this study, Swiss male mice were intraperitoneally administered with titanium dioxide (TiO2 ) and zinc oxide (ZnO) nanoparticles (NPs) and their mixture (1:1) at doses between 9.38 and 75 mg/kg for 5 weeks to evaluate reproductive toxicity. Both NPs and their mixture significantly (p < .001) altered sperm motility, reduced sperm numbers and increased abnormalities, while their mixture induced more sperm abnormalities than either TiO2 NPs or ZnO NPs. Both NPs and their mixture significantly (p < .05) reduced the LH level, while ZnO NPs alone and their mixture (p < .001) increased the testosterone levels at tested doses. The testes of exposed mice showed pathological changes and altered histomorphometrics. TiO2 NPs and ZnO NPs individually induced a significant (p < .01) reduction in SOD and CAT activities, while the mixture significantly (p < .001) decreased CAT activity and increased SOD activity. TiO2 NPs alone at 9.38 mg/kg induced a significant (p < .001) reduction in the GSH level, while both NPs and their mixture increased the MDA level significantly (p < .05). The data showed that the mixture had a synergistic interaction to induce testicular damage. Overall, oxidative stress may be involved in the NP-mediated testicular damage observed.

Reproductive toxicity assessment of Olusosun municipal landfill leachate in Mus musculus using abnormal sperm morphology and dominant lethal mutation assays.

Mixture of xenobiotics in Olusosun landfill leachates (OSL) induced somatic and germ cell genotoxicity in eukaryotic cells. However, whether the DNA-damaged germ cells can increase reproductive failure, embryotoxicity and/or teratogenicity during fertilization remained unclear. The study herein investigated reproductive toxicity and embryotoxic effects of OSL in mice using abnormal sperm morphology and dominant lethal mutation assays. Six mice per group were exposed to 0.5 mL of 5, 10, 25, and 50% OSL (v/v, leachate/distilled water) for 5 consecutive days. Similar treatment was giving to distilled water and cyclophosphamide (20 mg/kg body weight), as negative and positive control groups, respectively. At 35 day post-treatment, the mice were sacrificed and examined for cauda epididymal abnormal sperm morphology, testicular weight change and histopathological lesions. Male mice were mated to untreated nulliparous females (1 treated male per 2 untreated females) until vaginal plugs were observed. Pregnant females were sacrificed on 14th day post-mating and uterine content examined for number of implants per female, resorptions, and live and dead fetuses. OSL significantly increased teratozoospermia by 31.02% with amorphous sperm head having the highest frequency and sperm with two heads, the least. Histology of the testes revealed congestion of the interstitial blood vessels, spermatid retention and disorganization of the germinal epithelium from the basal compartment. OSL insignificantly increased absolute and relative testes weights but reduced body weight of exposed mice. There was insignificant induction of dominant lethal mutation, although low fertility was observed in the treated mice. The analyzed physico-chemical parameters and heavy metals/metalloids in OSL were higher than permissible limits. The metals, physico-chemical parameters and other unanalyzed substances in OSL induced abnormal sperm morphology, altered testes pathology, but did not significantly induce dominant lethal mutation in mice. Constituents of OSL are toxic to the male reproductive organs in mice and suggests harmful impact on public health. Indiscriminate disposal of solid wastes in the environment should be prevented.

Experimental simulation of somatic and germ cell genotoxicity in male Mus musculus fed extracts of lead contaminated Pleurotus ostreatus (white rot fungi).

Exposure to lead (Pb) is a major risk factor in reproductive toxicity, somatic, and germ cell genotoxicity. Exposure via deteriorating Pb paints and contaminated air, soil, and water had been the primary routes. However, with increasing reports of Pb accumulation in mushrooms and other food items may increase the etiology of Pb poisoning. The study herein investigated somatic genotoxicity and reproductive abnormalities in mice fed extracts of Pb-contaminated Pleurotus ostreatus. Male mice were fed aqueous extracts of P. ostreatus cultivated in 0, 10, 20, 100, 200, 500, and 1000 mg/L of Pb-contaminated rice straw for 35 days. Testes were analyzed for Pb accumulation, histopathology and relative weight gain, caudal epididymis for abnormal sperm morphology, and bone marrow for micronucleus test. Concentration-related significant increase in Pb accumulation was observed in P. ostreatus and testes of exposed mice. Decrease testicular weight, congestion of blood vessels, necrosis, and disorganization of the seminiferous tubules were observed in treated mice. In addition, fold increase of 2.78, 3.39, 6.67, 7.21, 9.63, and 9.70 in abnormal sperm morphology in accordance with the Pb concentrations respectively, confirmed reproductive toxicity. Significant increase in micronucleated polychromatic (PCE) and normochromatic (NCE) erythrocytes and concentration-related decrease PCE-NCE ratio in the bone marrow of treated mice suggest genome instability. Pb-contaminated P. ostreatus increased somatic and germ cell genotoxicity in mice. This may predispose the mice to genetic related syndromes and reproductive syndromes. It further suggests caution in the consumption of metal laden wild mushrooms and crop plants.

Experimental modeling of the acute toxicity and cytogenotoxic fate of composite mixtures of chromate, copper and arsenate oxides associated with CCA preservative using Clarias gariepinus (Burchell 1822).

Concurrent occurrence of chromium (Cr), copper (Cu) and arsenic (As) from chromated copper arsenate (CCA) wood preservative in aquatic ecosystems demands that their joint-actions in eliciting toxic effects be assessed for adequate understanding of the health risk they may pose to biota. Clarias gariepinus was exposed to As2O3 , CrO3 and CuO and their composite mixtures (1:1 and 1:1:1) at various concentrations (0 - 600 mg/L) for 96-h to determine the acute toxicity using OECD (1992) protocol. C. gariepinus was then exposed to sub-lethal concentrations corresponding to 6.25, 12.5, 25.0, 50.0 and 100% of the 96-h LC50 for 7 days to assess the cytogenotoxic effects using piscine micronucleus (MN) test. The 96-h LC50 showed that the metals/metalloid demonstrated differential interactions in a concentration dependent pattern. The 96-h LC50 showed that Cr was the most toxic while Cu and As:Cu were indeterminate (Cr > Cr:Cu > As:Cr > As > As:Cr:Cu > Cu = As:Cu indeterminate). Isobologram and synergistic ratio (SR) models predicted antagonistic interaction between Cu:Cr and As:Cr and synergism between As:Cu in the causation of morbidity and mortality of C. gariepinus. Interaction factor model predicted antagonism as common interactive mechanism among the metal/metalloid mixtures in the induction of MN and abnormal nuclear erythrocytes in C. gariepinus. Predicted interactions among the three metals/ metalloid were largely antagonism and synergism towards the induction of acute toxicity and cytogenotoxicity. The models employed herein may be useful in establishing environmental safe limits for mixtures of metals/metalloids against the induction of acute toxicity and DNA damage in lower aquatic vertebrates.

Genetic and systemic toxicity induced by silver and copper oxide nanoparticles, and their mixture in Clarias gariepinus (Burchell, 1822).

Unanticipated increase in the use of silver (Ag) and copper oxide (CuO) nanoparticles (NPs) due to their antimicrobial properties is eliciting environmental health concern because of their coexistence in the aquatic environment. Therefore, we investigated the genetic and systemic toxicity of the individual NPs and their mixture (1:1) using the piscine micronucleus (MN) assay, haematological, histopathological (skin, gills and liver) and hepatic oxidative stress analyses [malondialdehyde (MDA), reduced glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT)] in the African mud catfish, Clarias gariepinus. The fish were exposed to sublethal concentrations (6.25-100.00 mg/L) of each NP and their mixture for 28 days. Both NPs and their mixture induced significant (p < 0.05) increase in MN frequency and other nuclear abnormalities. There was significant decrease in haemoglobin concentration, red and white blood cell counts. Histopathological lesions observed include epidermal skin cells and gill lamellae hyperplasia and necrosis of hepatocytes. The levels of MDA, GSH and activities of SOD and CAT were impacted in C. gariepinus liver following the exposure to the NPs and their mixture. Interaction factor analysis of data indicates antagonistic genotoxicity and oxidative damage of the NPs mixture. These results suggest cytogenotoxic effects of Ag NPs, CuO NPs and their mixture via oxidative stress in Clarias gariepinus.

Exposure to effluent from pharmaceutical industry induced cytogenotoxicity, hematological and histopathological alterations in Clarias gariepinus (Burchell, 1822).

Pharmaceutical effluents contain toxic xenobiotics capable of contaminating aquatic environments. Untreated effluents are illegally discharged into aquatic environment in most developing countries. Pharmaceutical effluent induced alterations in biomarkers of genetic and systemic damage on rodents. However, information is relatively scarce on the possible cytogenotoxicity and systemic toxicity of this effluent on aquatic vertebrates. The study herein assessed the cytogenotoxic, hematological and histopathological alterations induced by pharmaceutical effluent in Clarias gariepinus. 96 h acute toxicity of the effluent was determined after C. gariepinus was exposed to six different concentrations (10 - 60 %) of the effluent. Subsequently, fish was exposed to sub-lethal concentrations (2.18 - 17.41 %) obtained from the 96 h LC50 for 7 and 14 days after which micronucleus (MN) and nuclear abnormalities (NAs) in peripheral erythrocytes were assessed as cytogenotoxic biomarkers, alterations in hematological indices and histopathological lesions were also examined. Fish, concurrently exposed to dechlorinated tap water and benzene (0.01 mL/L), served as negative and positive controls respectively. The derived 96 h LC50 of 17.41 % which was 1.89 times more toxic than the 24 h LC50 (32.95 %) showed that the effluent induced concentration-dependent mortality according to exposure duration. The effluent caused significant (p<0.05) time-dependent increase in the frequency of MN and abnormal nuclear erythrocytes compared to the negative control. Also, there was decrease in total erythrocyte counts, hemoglobin and hematocrit concentrations and increase in leucocyte and lymphocyte counts. The effluent induced pathological lesions on gills, liver and kidneys of treated fish. Higher physicochemical parameters than standard permissible limits in the effluent are capable of inducing genomic instability and systemic damage in fish. Pharmaceutical effluent can increase micropollutants in aquatic environmental and health risks to aquatic biota. There is need to promulgate stringent laws against illegal discharge of effluents into aquatic environment.

Intraperitoneal sodium metavanadate exposure induced severe clinicopathological alterations, hepato-renal toxicity and cytogenotoxicity in African giant rats (Cricetomys gambianus, Waterhouse, 1840).

Pollution of environment due to increased exploitation of minerals has been on the rise, and vanadium, a metal in the first transition series essential for mammalian existence, is a major component of air pollution. This study investigated the clinico-pathological, hepato-renal toxicity, and cytogenotoxicity of intraperitoneal exposure of African giant rats (AGRs), a proposed model for ecotoxicological research to sodium metavanadate. A total of 27 adult male African giant rats weighing 975 ± 54.10 g were distributed into two major groups: sodium metavanadate (SMV) treated and control. They were observed daily for clinical signs of toxicity. Four rats from each group were randomly collected and sacrificed after 3, 7, and 14 days of SMV treatment. Liver, kidney, and bone marrow were analyzed for histopathology and micronucleated normochromated and polychromated erythrocytes (MNNCE and MNPCE), respectively. Clinical signs in treated AGR include sluggish and weak movements, un-groomed fur, and labored breathing. Histology of the kidney revealed severe glomerular atrophy, tubular ectasia, and vacuolar degeneration of tubular epithelium, while liver histology showed sinusoidal congestion and severe hepatocellular necrosis after 14 days SMV exposure. Also, MNNCE and MNPCE significantly increased with a decrease in PCE/NCE ratio in SMV-treated AGR, suggestive of alternations in bone marrow cell proliferation. Hence, SMV treatment to AGR resulted to severe clinicopathologic alterations, kidney, and liver dysfunction and cytogenotoxicity evident by somatic mutation induction which could be severe with prolonged exposure. This suggests African giant rat as an ecotoxicological model to measure major health risks to animals and human populations in highly polluted environment.

Dietary ascorbic acid reduced micronucleus and nuclear abnormalities in Clarias gariepinus (Burchell 1822) exposed to hospital effluent.

Hospital effluents contain myriad of mutagens and genotoxins capable of increasing DNA damage in aquatic biota. African mudfish, Clarias gariepinus, are exposed to genotoxins when cultured in swamps and derelict water bodies often contaminated by effluents. Moreover, its DNA is susceptible to xenobiotic-induced lesions since it lacks L-gulonolactone oxidase and hence cannot synthesize L-ascorbic acid. This study investigated 96-h acute toxicity and protective effects of dietary ascorbic acid (AA) against micronucleus (MN) and abnormal nuclear (NAs) formation in C. gariepinus exposed to sub-lethal concentrations of hospital effluent. Six concentrations (0.5-3.0%) of the effluent were selected to determine the 96-h acute toxicity of the effluent in C. gariepinus, after range finding test. Fish were exposed to sub-lethal concentrations (0.08-1.30%) of the 96 h LC50. Two other groups were exposed to the 96 h LC50 (1.30%) of the effluent +50 and +100 mg/kg of dietary ascorbic for 7 days, and MN and NAs assessed in peripheral erythrocytes. The 96 h LC50 (1.30%) was 1.18 times more toxic than the 24 h LC50 (1.54%), indicating that the toxicity of the effluent increased with exposure duration. MN, nuclear bud, enucleated, fragmented nucleus (apoptosis), and necrotic erythrocytes significantly increase in effluent treated fish. Dietary AA reduced MN from 6.35-fold (1.30% treated group) to 3.72-fold (1.30% + 50 mg AA) and 3.54-fold (1.30% + 100 mg AA). Also, AA reduced total NAs from 2.26-fold (1.30%) to 1.40-fold (1.30% + 50 mg AA) and 1.06-fold (1.30% + 100 mg AA) compared to the control. Heavy metals and physicochemical parameters analyzed in the tested effluent possibly induced the mortality and cytogenotoxicity in C. gariepinus, and this was ameliorated by dietary AA.

Chemical characterization of simulated landfill soil leachates from Nigeria and India and their cytotoxicity and DNA damage inductions on three human cell lines.

Landfill soils are sources of emerging carcinogens, teratogens and mutagens in the environment. There is inadequate information on its possible health risk and cytogenotoxicity. This study evaluated chemical characterization of four simulated landfill leachates with their cytotoxicity and DNA damage in human cells. Hepatocarcinoma (HepG2), lymphoma (Jurkat) and osteosarcoma (HOS) cells, incubated with 6.25, 12.5, 25, 50, 75 and 100% of Aba Eku (AEL), Olusosun (OSL), Awotan (AWL) and Nagpur (NPL) simulated leachates for 24 h, were assessed for cell viability using MTT assay and morphological alterations. DNA damage was also assessed after 24 h treatment of cells with sub-lethal concentrations of the leachates using comet assay. Metals and organic compounds in the soil leachates were determined using inductively coupled plasma-mass spectrometry (ICP-MS) and gas chromatography-mass spectroscopy (GC-MS) respectively. The leachates induced significant cytotoxicity in the treated cells with evidence of apoptosis; shrunken morphologies, detachment from the substratum and cytoplasmic vacuolations. Similarly, there was significant DNA damage induced in the treated cells, with increased Olive tail moment, tail length and % tail DNA. Jurkat was the most sensitive (Jurkat > HepG2 > HOS) to the cytotoxic and genotoxic effects of the leachates. All the analyzed metals except Cd, Fe, Zn and Mn were found at levels lower than standard allowable limits. 32, 17, 23 and 23 different PAHs and PCBs were detected in AEL, AWL, OSL and NPL respectively, at varying retention peak times. These toxic constituents induced the observed cytogenotoxicity in the cells and may suggest possible public health risk.

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.

In vivo micronucleus test in the assessment of cytogenotoxicity of landfill leachates in three animal models from various ecological habitats.

The in vivo micronucleus (MN) test, a standard test for the genotoxicity screening of xenobiotics, was used to evaluate the cytotoxic and genotoxic activities of landfill leachates in Clarias gariepinus, Coturnix coturnix japonica and Rattus norvegicus. These organisms were exposed to various sub-lethal concentrations (1-50%) of Olusosun and Aba Eku landfill leachates. At post exposure, peripheral erythrocytes from catfish and quail, and bone marrow cells of quail and rat were subjected to MN analysis following standard protocols. The leachates induced significant increase in MN formation and total nuclear abnormalities (NAs) in the peripheral erythrocytes of catfish and quail. NAs occurred in the order; BN > BL > LB > NT in the catfish and BN > BudN > TLN > TN in quail. There was significant increase in MN formation in the bone marrow cells of quail, and micronucleated polychromatic erythrocytes and micronucleated normochromatic erythrocytes formation in the bone marrow of rats. The concentration dependent significant (p < 0.05) decrease in the PCE/NCE ratio in the bone marrow of the leachate treated rats suggest alterations in the bone marrow cell proliferation, leading to the suppression of immature erythrocytes (PCE). MN induction showed positive corrections with leachate concentrations in the test organisms; and it increased with exposure duration in the catfish. Indiscriminate disposal of solid waste generates leachates containing multiple xenobiotics that are capable of increasing genomic instability among vertebrates inhabiting various ecological habitats.

Correlation of melanophore index with a battery of functional genomic stress indicators for measurement of environmental stress in aquatic ecosystem.

The correlation of primary stress indicator; melanophore index (MI) with set of genomic stress indicators is important for a better understanding of the cellular stress pathway induced by xenobiotics in aquatic species. This study presents a correlation between melanophore index (MI) and genomic stress indicators in Oreochromis mossambicus treated with lead nitrate, phenol and hexachlorocyclohexane (HCH). O. mossambicus was exposed to sub-lethal concentrations of the different LC50 values (96 h) of the tested chemicals at varying exposure periods and the response via genomic stress indicators and scale melanophores were assessed in accordance with standard protocols. Melanophore index decreased significantly (p<0.01) in a time dependent pattern to the tested chemicals. Gene expression showed significant time dependent increase in the expression of heat shock proteins (HSP70 and HSP60). Vitellogenin (Vtg) expression insignificantly altered. Significant increase in the expression of melanin concentrating hormone (MCH) was observed in response to hexachlorocyclohexane (HCH) in the treated fish. The findings demonstrated an inverse relationship between melanophore index and the set of genomic stress indicators.

The genotoxicity and systemic toxicity of a pharmaceutical effluent in Wistar rats may involve oxidative stress induction.

There is scarcity of information on the possible mechanisms of pharmaceutical effluent induced genotoxicity and systemic toxicity. This study investigated the genotoxicity and systemic toxicity of a pharmaceutical effluent in Wistar rats. Rats were orally treated with 5-50% concentrations of the effluent for 28 days. At post-exposure, blood, liver, kidney and bone marrow cells were examined for alterations in serum biochemical parameters and hematological indices, histopathological lesions and micronucleated polychromatic erythrocytes formation (MNPCE). The effluent caused concentration independent significant (p < 0.05) alterations in aspartate (AST) and alanine (ALT) aminotransferases, superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA), total and direct bilirubin and creatinine. There was reduction in red blood count (RBC), hemoglobin concentration (HGB), platelets, percentage hematocrit (HCT), white blood count (WBC) and mean corpuscle hemoglobin (MCH) except mean corpuscle hemoglobin concentration (MCHC), which increased in the treated rats. Histopathological lesions observed in the liver and kidney of the effluent treated rats were thinning of the hepatic cord, kuffer cell hyperplasia, vacuolation of the hepatocytes and renal cells, multifocal inflammatory changes, necrosis and congestion of the renal blood vessels and central vein. MNPCE significantly increase in the bone marrow of the treated rats compared to the negative control. The concentration of some toxic metals and anions in the effluent were above standard permissible limits. These findings showed that the pharmaceutical effluent caused somatic DNA damage and systemic toxicity in rats may involve induction of oxidative stress, suggesting environmental contamination and health risks in wildlife and humans.

Hospital waste incinerator bottom ash leachate induced cyto-genotoxicity in Allium cepa and reproductive toxicity in mice.

The potentials of hospital incinerator bottom ash leachate (HIBAL) to induce cyto-genotoxicity in Allium cepa and reproductive anomalies in the mouse were investigated. The leachate obtained from simulation of the bottom ash was analyzed for some physico-chemical parameters. The A. cepa, mouse sperm morphology and histopathological tests were carried out at concentrations ranging from 1% to 50% of the leachate sample. In A. cepa, HIBAL caused significant (p < 0.05) inhibition of root growth and induction of chromosomal aberrations. In the animal assays, there was 100% mortality at the 50% concentrations. The leachate caused insignificant (p > 0.05) concentration-dependent induction of various types of sperm morphology. There was accumulation of fluid in the seminiferous tubule lumen and necrosis of stem cells in the testes. These effects were believed to be provoked by the somatic and germ cell genotoxins, particularly the heavy metals in the leachate. Our finding is of environmental and public health significance.