Physiological and histopathological alterations in male Swiss mice after exposure to titanium dioxide (anatase) and zinc oxide nanoparticles and their binary mixture.

Existing studies have shown the systemic damage of titanium dioxide (TiO2) or zinc oxide (ZnO) nanoparticles (NPs), but there is little or no existing knowledge on the potential adverse toxic effects of the mixture of the two. In order to investigate the in vivo toxic effect of the mixture of TiO2 NPs and ZnO NPs, the acute toxicities of TiO2 NPs, ZnO NPs by themselves, and their mixture (1:1) were determined. The systemic toxicities of the individual NPs and mixture were evaluated in mice using hematological indices, hepatic, renal, and lipid profile parameters, and histopathology as endpoints. NPs were intraperitoneally administered at doses of 9.38, 18.75, 37.50, 75.00, and 150.00 mg/kg bw each. Individual NPs and their mixture were administered daily for 5 and 10 d, respectively. The LD50 of ZnO NPs was 299.9 mg/kg while TiO2 NPs by themselves or TiO2 NPs + ZnO NPs were indeterminate due to the absence of mortality of the male mice treated. TiO2 NPs, ZnO NPs by themselves and TiO2 NPs + ZnO NPs induced significant alterations in the hematological and biochemical parameters, with higher toxicity at 10 d. Histopathological lesions were observed in the liver, kidneys, spleen, heart, and brain of mice treated with the individual NPs and their mixture. TiO2 NPs + ZnO NPs were able to induce a higher systemic toxicity than TiO2 NPs or ZnO NPs individually. Our data suggest that more comprehensive risk assessments should be carried out on the mixture of NPs before utilization in consumer products.

In vitro cytotoxicity of co-exposure to superparamagnetic iron oxide and solid lipid nanoparticles.

Increased production and use of different types of nanoparticles (NPs) in the last decades has led to increased environmental release of these NPs with potential detrimental effects on both the environment and public health. Information is scarce in the literature on the cytotoxic effect of co-exposure to many NPs as this concern is relatively recent. Thus, in this study, we hypothesized scenarios of cell's co-exposure to two kinds of NPs, solid lipid nanoparticles (SLNs) and superparamagnetic iron oxide nanoparticles (SPIONs), to assess the potential cytotoxicity of exposure to NPs combination. Cytotoxicity of SPIONs, SLNs, and their 1:1 mixture (MIX) in six tumor and six non-tumor cell lines was investigated. The mechanisms underlining the induced cytotoxicity were studied through cell cycle analysis, detection of reactive oxygen species (ROS), and alterations in mitochondrial membrane potential (ΔΨM). Double staining with acridine orange and ethidium bromide was also used to confirm cell morphology alterations. The results showed that SPIONs induced low cytotoxicity compared to SLNs. However, the mixture of SPIONs and SLNs showed synergistic, antagonistic, and additive effects based on distinct tests such as viability assay, ROS generation, ΔΨM, and DNA damage, depending on the cell line. Apoptosis triggered by ROS and disturbances in ΔΨM are the most probable related mechanisms of action. As was postulated, there is possible cytotoxic interaction between the two kinds of NPs.

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.

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.

Evidence of a multiple insecticide resistance in the malaria vector Anopheles funestus in South West Nigeria.

BACKGROUND: Knowing the extent and spread of insecticide resistance in malaria vectors is vital to successfully manage insecticide resistance in Africa. This information in the main malaria vector, Anopheles funestus sensu stricto, is completely lacking in the most populous country in Africa, Nigeria. This study reports the insecticide susceptibility status and the molecular basis of resistance of An. funestus as well as its involvement in malaria transmission in Akaka-Remo, a farm settlement village in southwest Nigeria. RESULTS: Plasmodium infection analysis using TaqMan protocol coupled with a nested PCR revealed an infection rate of 8% in An. funestus s.s. from Akaka-Remo. WHO susceptibility tests showed this species has developed multiple resistance to insecticides in the study area. Anopheles funestus s.s. population in Akaka-Remo is highly resistant to organochlorines: dieldrin (8%) and DDT (10%). Resistance was also observed against pyrethroids: permethrin (68%) and deltamethrin (87%), and the carbamate bendiocarb (84%). Mortality rate with DDT slightly increased (from 10 to 30%, n = 45) after PBO pre-exposure indicating that cytochrome P450s play little role in DDT resistance while high mortalities were recorded after PBO pre-exposure with permethrin (from 68 to 100%, n = 70) and dieldrin (from 8 to 100%, n = 48) suggesting the implication of P450s in the observed permethrin and dieldrin resistance. High frequencies of resistant allele, 119F in F0 (77%) and F1 (80% in resistant and 72% in susceptible) populations with an odd ratio of 1.56 (P = 0.1859) show that L119F-GSTe2 mutation is almost fixed in the population. Genotyping of the A296S-RDL mutation in both F0 and F1 samples shows an association with dieldrin resistance with an odd ratio of 81 (P < 0.0001) (allelic frequency (R) = 76% for F0; for F1, 90 and 10% were observed in resistant and susceptible populations, respectively) as this mutation is not yet fixed in the population. CONCLUSION: The study reports multiple insecticide resistance in An. funestus from Akaka Remo. It is, therefore, necessary to pay more attention to this major malaria vector for effective malaria control in Nigeria.

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.

Electronic waste leachate-mediated DNA fragmentation and cell death by apoptosis in mouse fibroblast (NIH/3T3) cell line.

This study investigated the apoptotic effect of electronic waste on fibroblast cell line. Cells were treated with different concentrations of the leachate for 24h. Cell viability was detected by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) test, nuclear morphology of cells was explored by acridine orange (AO)/ethidium bromide (EB) double staining, mitochondrial membrane potential was evaluated using JC-1 probe while cell cycle analysis was conducted using flow cytometry. The oxidative status was detected using DCFH-DA (dichlorofluorescin diacetate) probe and the relationship between cell death and ROS (reactive oxygen species) was investigated using N-acetylcysteine. Results showed an increased cell death as detected by MTT assay and AO/EB staining. Cell cycle analysis indicated an induction of sub/G1 events while JC-1 probe showed significant disruption of mitochondrial membrane potential. There was significant induction of ROS, while N-acetylcysteine protected the cells from DNA damage. These suggest apoptotic pathway as a possible mechanism of e-waste induced cyto-genotoxicity.

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.

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.

The first-line antituberculosis drugs, and their fixed-dose combination induced abnormal sperm morphology and histological lesions in the testicular cells of male mice.

Rifampicin (RIF), Isoniazid (INH), Ethambutol (EMB), Pyrazinamide (PZA), and/or their fixed-dose combination (FDC) are extensively prescribed in the cure of Tuberculosis (TB) globally. In spite of the beneficial effect, these drugs are capable of inducing cellular toxicity. Existing information on the genotoxic effects of the first-line anti-TB drugs is limited and contentious. Herein, we evaluated the reproductive genotoxicity of RIF, INH, EMB, PZA, and their FDC utilizing the mouse sperm morphology assay. Histological examination of the testes of exposed mice was also performed. Male Swiss albino mice (11-13 weeks old) were intraperitoneally exposed for 5 consecutive days to each of the anti-TB drugs at four different doses of 6.25, 12.5, 25, and 50 mg/kg bw of PZA; 2.5, 5.0, 10, and 20 mg/kg bw of RIF; 1.25, 2.5, 5.0 and 10 mg/kg bw of INH; 3.75, 7.5, 15 and 30 mg/kg bw of EMB; and 7, 14, 28 and 56 mg/kg bw of FDC corresponding respectively to ×0.25, ×0.5, ×1 and ×2.0 of the standard daily dose. In comparison with the negative control (normal saline), there was no significant difference in the testicular weight and organo-somatic index of exposed mice. There was an increase (p > 0.05) in the frequency of abnormal spermatozoa at most of the tested doses of each drug and a dose-dependent decrease with the FDC. Each of the anti-TB drugs except the FDC induced pathological lesions in the testes. These findings suggest that the individual first-line anti-TB drug unlike the FDC has the potential to provoke testicular anomalies in male mice.

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.

Exposure to a contaminated tropical freshwater (Awba Dam) in Ibadan, Nigeria, induced cytogenotoxicity and haemato-pathological changes in Clarias gariepinus.

Awba reservoir serves the purpose of water supply in the University of Ibadan, Nigeria. Recent reports on pollution status have focused on toxicological implication of contaminants in this reservoir. But none is on genetic and systemic toxicity of the water in fish. We investigated cytogenotoxicity of Awba Dam water (ADW) on Clarias gariepinus using piscine micronucleus (MN) assay. Haematological and histopathological changes were also evaluated. Bi-monthly composite water samples were collected from the reservoir from July to October, 2018. The water was used to cultivate juvenile C. gariepinus in the laboratory for 1-4 months, and tap water was used as the negative control. Peripheral blood erythrocytes from the caudal veins of C. gariepinus were used for the monthly MN assessment. There was significant (p < 0.05) induction of MN and other erythrocyte nuclear abnormalities in C. gariepinus within the period of study. There were variations in the haematological indices and pathological alterations in sections of the gill, liver and kidney of C. gariepinus. The levels of some heavy metals in ADW were above standard permissible limits and might have contributed to the observed cytogenetic and systemic disruptions. These findings may be used by the concerned authorities to evolve management strategies for the reservoir's health and biologic resources.

Environmental contamination and public health effects of electronic waste: an overview.

PURPOSE: In recent years, electronic waste has become the fastest growing waste stream globally with potential deleterious environmental and public health effects from its hazardous constituents. This review aims at providing an up-to-date information on the environmental and public health effects of e- wastes, and also identify research gaps that could form basis of further innovative studies on this important subject. METHODS: We carried out literature survey using several search engines. All available literature which reported directly on environmental contamination of air, soil, and water by e-wastes, and their effects on exposed plants, animals, and humans were used in other to generate an updated information. RESULTS: High production volume coupled with indiscriminate disposal and informal recycling has made electronic waste (e-waste) to become a global public and environmental health issue. E-waste is made up of different hazardous substances such as heavy metals and persistent organic pollutants with the capacity to contaminate the environment if processed or recycled inappropriately. Humans and animals become exposed to e-waste constituents via ingestion, inhalation, and dermal contact. Several health effects have been linked to e-wastes. The most susceptible were children, pregnant women, and workers in primitive recycling sites. Generation of e-waste is predicted to increase drastically in the next decade with the potential complex interactive effects of its constituents. CONCLUSION: This review is an up-to-date assessment of studies and reports on e-waste environmental contamination and public health effects. The review has shown that e-waste contains constituents that caused adverse environmental effects and toxicity to the biota. However, there is an enormous data gap between exposure quantification and possible health effects. More studies are needed to elucidate and provide holistic information on environmental and public health dangers posed by e-waste constituents.

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.

Elevated Serum Pb, Ni, Cd, and Cr Levels and DNA Damage in Exfoliated Buccal Cells of Teenage Scavengers at a Major Electronic Waste Dumpsite in Lagos, Nigeria.

This study investigated the levels of Pb, Ni, Cd, and Cr in the blood, and DNA damage in exfoliated buccal cavity of scavenging teenagers at Alaba International electronic market, Nigeria. One hundred and ninety-nine blood samples were collected from scavenging teenagers in Alaba International market (n = 95) and the control group (n = 104). Blood Pb, Ni, Cd, and Cr concentrations were analyzed using graphite furnace atomic absorption spectrometry. Genotoxicity assessment was through exfoliated buccal cell micronucleus assay, while questionnaire was used to collect data on factors that can contribute to the blood heavy metal levels. The blood Pb of scavenging teenagers from Alaba International market varied from 2.84 to 189.44 µg/L compared to the control group with a range of 0.01 to 4.21 µg/L. The blood Cd (range = 0.38-12.35 µg/L), Cr (range = 0.20-19.03 µg/L), and Ni (range = 0.24-21.01 µg/L) were higher significantly in the Alaba group compared to the control group (Cd: range = 0.00-0.96 µg/L; Cr: range = 0.01-0.72 µg/L; and Ni: range = 0.00-0.28 µg/L). Frequencies of micronucleus, karyolysis, and pycnosis in the exfoliated buccal cells in Alaba group also increased significantly compared with the control group. Spearman correlation analyses indicated a positive correlation between the blood Pb, Ni, Cd, and Cr and induced micronucleus in the buccal exfoliated cells. Furthermore, data showed a positive correlation between Pb, Ni, Cd, and Cr and the duration of scavenging of the teenagers. However, a negative correlation was observed between educational status and Pb, Ni, Cd, and Cr. E-waste indiscriminate disposal and primitive recycling processes may be responsible for the high blood heavy metal levels, and consequently an increased DNA damage in exfoliated buccal cells in scavenging teenagers at Alaba International market. Graphical Abstract.

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.

Haematological Studies and Micronucleus Assay of Straw-Coloured Fruit Bats (Eidolon helvum).

The straw-coloured fruit bats (Eidolon helvum) are the most widely distributed megachiropteran species in Africa. Studies have shown that they migrate, and are likely exposed to environmental pollutants across population. This study was designed to investigate genotoxicity via the bone marrow micronucleus assay and haematological alterations of Eidolon helvum in the tropics. Healthy straw-coloured fruit bats (Eidolon helvum; n=20) were captured from two geographical regions, Ogun and Gombe States in Nigeria and were grouped based on sex and age.  Blood samples were collected for haematology and osmotic fragility, and bone marrow samples for genotoxicity studies. Results showed no significant differences in erythrocytes and leucocytes values across age and sex. The erythrocytes osmotic fragility was higher in juvenile than in adults at 0 and 0.1%NaCl, while it was higher in adult males than in adult females at 0 and 0.3% NaCl. The erythrocytes and leucocytes parameters in straw colored fruit bats were within the reference values seen in literature except the higher monocyte counts suggesting chronic inflammation. There were increased levels of micronucleated polychromatic erythrocytes and normochromatic erythrocytes in the straw-coloured fruit bats indicating genotoxicity and cytotoxicity, respectively. The present study provided baseline research data on the haematology and micronucleus profile of the straw-coloured fruit bats in Nigeria. This is perhaps the first study on haematology and micronucleus assay of in straw-colored fruit bats in the tropics.

Investigating the molecular basis of multiple insecticide resistance in a major malaria vector Anopheles funestus (sensu stricto) from Akaka-Remo, Ogun State, Nigeria.

BACKGROUND: Understanding the mechanisms used by Anopheles mosquitoes to survive insecticide exposure is key to manage existing insecticide resistance and develop more suitable insecticide-based malaria vector control interventions as well as other alternative integrated tools. To this regard, the molecular basis of permethrin, DDT and dieldrin resistance in Anopheles funestus (sensu stricto) at Akaka-Remo was investigated. METHODS: Bioassays were conducted on 3-5-day-old adult An. funestus (s.s.) mosquitoes for permethrin, DDT and dieldrin susceptibility test. The molecular mechanisms of mosquito resistance to these insecticides were investigated using microarray and reverse transcriptase PCR techniques. The voltage-gated sodium channel region of mosquitoes was also screened for the presence of knockdown resistance mutations (kdr west and east) by sequencing method. RESULTS: Anopheles funestus (s.s.) population was resistant to permethrin (mortality rate of 68%), DDT (mortality rate of 10%) and dieldrin (mortality rate of 8%) insecticides. Microarray and RT-PCR analyses revealed the overexpression of glutathione S-transferase genes, cytochrome P450s, esterase, trypsin and cuticle proteins in resistant mosquitoes compared to control. The GSTe2 was the most upregulated detoxification gene in permethrin-resistant (FC = 44.89), DDT-resistant (FC = 57.39) and dieldrin-resistant (FC = 41.10) mosquitoes compared to control population (FC = 22.34). The cytochrome P450 gene, CYP6P9b was also upregulated in both permethrin- and DDT-resistant mosquitoes. The digestive enzyme, trypsin (hydrolytic processes) and the cuticle proteins (inducing cuticle thickening leading to reduced insecticides penetration) also showed high involvement in insecticide resistance, through their overexpression in resistant mosquitoes compared to control. The kdr east and west were absent in all mosquitoes analysed, suggesting their non-involvement in the observed mosquito resistance. CONCLUSIONS: The upregulation of metabolic genes, especially the GSTe2 and trypsin, as well as the cuticle proteins is driving insecticide resistance of An. funestus (s.s.) population. However, additional molecular analyses, including functional metabolic assays of these genes as well as screening for a possible higher cuticular hydrocarbon and lipid contents, and increased procuticle thickness in resistant mosquitoes are needed to further describe their distinct roles in mosquito resistance.

Genotoxicity assessment of a pharmaceutical effluent using four bioassays.

Pharmaceutical industries are among the major contributors to industrial waste. Their effluents when wrongly handled and disposed of endanger both human and environmental health. In this study, we investigated the potential genotoxicity of a pharmaceutical effluent, by using the Allium cepa, mouse- sperm morphology, bone marrow chromosome aberration (CA) and micronucleus (MN) assays. Some of the physico-chemical properties of the effluent were also determined. The A. cepa and the animal assays were respectively carried out at concentrations of 0.5, 1, 2.5, 5 and 10%; and 1, 5, 10, 25 and 50% of the effluent. There was a statistically different (p < 0.05), concentration-dependent inhibition of onion root growth and mitotic index, and induction of chromosomal aberrations in the onion and mouse CA test. Assessment of sperm shape showed that the fraction of the sperm that was abnormal in shape was significantly (p < 0.05) greater than the negative control value. MN analysis showed a dose-dependent induction of micronucleated polychromatic erythrocytes across the treatment groups. These observations were provoked by the toxic and genotoxic constituents present in test samples. The tested pharmaceutical effluent is a potentially genotoxic agent and germ cell mutagen, and may induce adverse health effects in exposed individuals.

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.