Genotoxicity of Chlorpyrifos and the Antimutagenic Role of Lettuce Leaves in Male Mice

Chlorpyrifos [O O-diethyl-O-(3 5 6-trichloro-2-pyridyl)-phosphorothioate] is one of the most widely used organophosphate insecticides. Previous studies proved that chlorpyrifos, at different doses, induced genotoxicity. In Egyptian foods, the residual levels of pesticides are often higher than those found in developed country ones. So the aim of this research was to evaluate the genotoxicity of the insecticide chlorpyrifos at doses equal to its maximum residue limit (MRL) in the leafy vegetables, its double and quadruple (0.5, 1 and 2 mg/kg body weight) in somatic and germ cells of male mice. In addition to that, evaluating the role of lettuce leaves as antigenotoxic in reducing the genotoxic effects of chlorpyrifos tested doses when concurrently administrated to these animals. The study was conducted on adult male laboratory mice at three levels: bone marrow cells as a model for mitotic chromosome aberrations, spermatocytes as a model for meiotic chromosomes and sperm count and morphology. The results of the present study indicate that the treatment of male mice with chlorpyrifos by oral gavages for three months induced significant increase in the frequencies of total chromosomal aberrations in both somatic and germ cells in relation to control groups. Results of the sperm analysis showed that chlorpyrifos induced significant decrease in the sperm count when compared to negative control. Furthermore, it induced significant increase in head and tail sperm abnormalities, among which coiled tail was considered the most obvious sperm abnormality induced by chlorpyrifos. At the same time, the present study indicated that lettuce leaves feed concurrently with three doses of chlorpyrifos could not protect cells from damage.Chlorpyrifos [O O-diethyl-O-(3 5 6-trichloro-2-pyridyl)-phosphorothioate] is one of the most widely used organophosphate insecticides. Previous studies proved that chlorpyrifos, at different doses, induced genotoxicity. In Egyptian foods, the residual levels of pesticides are often higher than those found in developed country ones. So the aim of this research was to evaluate the genotoxicity of the insecticide chlorpyrifos at doses equal to its maximum residue limit (MRL) in the leafy vegetables, its double and quadruple (0.5, 1 and 2 mg/kg body weight) in somatic and germ cells of male mice. In addition to that, evaluating the role of lettuce leaves as antigenotoxic in reducing the genotoxic effects of chlorpyrifos tested doses when concurrently administrated to these animals. The study was conducted on adult male laboratory mice at three levels: bone marrow cells as a model for mitotic chromosome aberrations, spermatocytes as a model for meiotic chromosomes and sperm count and morphology. The results of the present study indicate that the treatment of male mice with chlorpyrifos by oral gavages for three months induced significant increase in the frequencies of total chromosomal aberrations in both somatic and germ cells in relation to control groups. Results of the sperm analysis showed that chlorpyrifos induced significant decrease in the sperm count when compared to negative control. Furthermore, it induced significant increase in head and tail sperm abnormalities, among which coiled tail was considered the most obvious sperm abnormality induced by chlorpyrifos. At the same time, the present study indicated that lettuce leaves feed concurrently with three doses of chlorpyrifos could not protect cells from damage.Genotoxidade de Chlorpyrifos e papel antimutagênico das folhas de alfaces em ratosResumoChlorpyrifos [O O-diethyl-O-(3 5 6-trichloro-2-pyridyl)-phosphorothioate] é um dos inseticidades a basede organofosfato mais utilizados. Estudos anteriores provaram que chlorpyrifos, em doses diferentes,induziu a genotoxidade. Em alimentos egípcios, os níveis residuais de pesticidas são normalmentemais elevados que aqueles encontrados em países desenvolvidos. Então, o objetivo dessa pesquisafoi o de avaliar a genotoxidade do inseticida chlorpyrifos em doses iguais ao limite residual máximo(MRL) em vegetais com folhas, seu dobro e quádruplo (0.5, 1 and 2 mg/kg peso corpóreo) emcélulas somáticas e germinativas de ratos. Além disso, avaliando o papel das folhas de alfacecomo antígenos tóxicos na redução dos efeitos de genotoxidade em doses de chlorpyrifos testadasquando administradas a esses animais. O estudo foi conduzido em ratos adultos de laboratório, emtrês níveis: células ósseas como modelo para aberrações miótico-cromossômicas; espermatócitoscomo modelo meiótico-cromossômico, contagem e morfologia de espermatozóides. Os resultadosdo presente estudo indicaram que o tratamento de ratos com chlorpyrifos, administrado emdoses orais por três meses induziram um significativo aumento da freqüência de aberraçõescromossômicas totais tanto em células somáticas quanto em células germinativas se relacionadosaos grupos de controle. Os resultados da análise de espermatozóide demonstraram que chlorpyrifosinduziu uma diminuição significativa na contagem de espermatozóides quando comparado aogrupo de controle negativo. Além disso, também houve a indução de um aumento significativoem anomalias na cabeça e na cauda dos espermatozóides, dentre as quais a cauda enroladafoi considerada a anomalia mais óbvia entre os espermatozóides induzidos por chlorpyrifos. Aomesmo tempo, o presente estudo indicou que as folhas de alface alimentadas regularmente comtrês doses de chlorpyrifos não conseguiram proteger suas células de danos.

Evaluation of the alleviative role of Chlorella vulgaris and Spirulina platensis extract against ovarian dysfunctions induced by monosodium glutamate in mice

Microalgae provide a wealthy natural resource of bioactive compounds, which have many biological activities. Monosodium glutamate is a food additive that acts either as food preservatives or as tastiness enhancer. It was confirmed that monosodium glutamate poses a serious responsibility in the pathogenesis of anovulatory infertility. Therefore, the idea of this research was directed to reveal efficiency of Chlorella vulgaris and Spirulina platensis extracts against the ovarian dysfunction resulted due to monosodium glutamate consumption. Adult female albino mice were gavages orally monosodium glutamate alone or with either Chlorella vulgaris or Spirulina platensis aqueous extracts for 28 days. Female mice were subjected to superovulation to study the oocytes nuclear maturation stages. Histological and quantitative investigation was carried on ovaries. Biochemical assessment to measure the sex hormones level and ovarian enzymatic antioxidants was done. In addition, ovarian antioxidant mRNA genes were determined using quantitative PCR and Glyceraldehyde-3-phosphate dehydrogenase was used as an internal control. The result revealed that monosodium glutamate reduced the oocytes quality and maturation rate, while, both algae improve the oocyte quality and maturation rate than in monosodium glutamate group. Chlorella vulgaris and Spirulina platensis improved the monosodium glutamate ovarian tissue histological alteration, sex hormones content and raised the ovarian enzymatic antioxidants level. In addition, monosodium glutamate markedly diminished the Glutathione peroxidase, superoxide dismutase and catalase mRNA expressions, However, Chlorella vulgaris or Spirulina platensis upregulated the expression of genes close to control. In conclusion, Chlorella vulgaris and Spirulina platensis showed potential alleviative role against the monosodium glutamate ovarian dysfunction. Keywords: Monosodium glutamate, Chlorella vulgaris, Spirulina platensis, Maturation rate, Sex hormones, Antioxidant enzymes, Antioxidant gene expression