Sperm Head Abnormality Assay Research Articles

Determination of genoprotection against cyclophosphamide induced toxicity in bone marrow of Swiss albino mice by Moringa oleifera leaves and Tinospora cordifolia stem

ABSTRACT The present study aimed to determine the genoprotective activity and safety of Moringa oleifera leave and Tinospora cordifolia stem extracts against cyclophosphamide (CP)-induced genotoxicity utilizing Swiss albino mice. Animals were divided into 14 groups for subacute treatment with either M. oleifera or T. cordifolia extracts daily for 28 days. The extract doses selected were 100, 200 or 400 mg/kg b.w administered orally alone or combined with CP (50 mg/kg b.w. intraperitoneally daily for 5 days). Analyses performed included the comet assay, micronucleus test (MN) in bone marrow cells and sperm head abnormality assay (SHA). M. oleifera and T. cordifolia extracts induced no significant genotoxic effects on somatic and germ cells. In contrast, for all cells examined M. oleifera and T. cordifolia extracts inhibited DNA damage initiated by CP. Taken together data demonstrated that both plant extracts did not exhibit marked genotoxic effects but displayed potential chemoprotective properties against CP-induced genotoxicity in Swiss mice.

Trichostatin A, an epigenetic modifier, mitigates radiation-induced androphysiological anomalies and metabolite changes in mice as evident from NMR-based metabolomics

Purpose: Ionizing radiation is known to damage male reproductive system. Current study aims to study the mitigative effects of trichostatin A on male reproductive system and accompanying metabolite changes in testicular tissue of mice.Materials and methods: Eight-week-old male C57 Bl/6J mice were exposed to 2 Gy γ-radiation with or without trichostatin A administration. The animals were sacrificed at various time intervals for organ body weight index, sperm head abnormality assay, sperm mobility assay, and study of various metabolites in testicular tissue using NMR spectroscopy.Results: Ionizing radiation induced no significant change in organ body weight index at any time points studied, however a significant increase in sperm head abnormality and significant decrease in sperm mobility was evident on fifth postirradiation week. trichostatin A administration, 1 and 24 h postirradiation, could efficiently mitigate radiation-induced changes studied. NMR metabolome profile also showed prominent changes associated with energy metabolism, osmolytes and membrane metabolism at 24 h postirradiation and some of these changes (choline, glycerolphosphoethanol amine, and glycine) were persistent till fifth postirradiation week. Trichostatin A administration resulted in reverting metabolic profile of the irradiated animals to normal level suggesting its mitigative role.Conclusion: Results obtained suggest that trichostatin A could restore normal metabolic profile of testicular tissue of irradiated male mice and also restored certain morphological and functional properties of sperms. Trichostatin A thus could further be exploited for its radio-mitigative properties.

An in vivo assay of the mutagenic potential of imidacloprid using sperm head abnormality test and dominant lethal test

Objective: To assess the mutagenic effects of imidacloprid in germ cells of Swiss albino male mice by sperm head abnormality (SHA) assay and dominant lethal test (DLT). Methods: Swiss albino mice were exposed to imidacloprid (22, 11 and 5.5 mg/kg/day) along with 3% gum acacia as vehicle control through oral route for 7, 14 and 28 days for SHA assay and for 28 days for DLT. The epididymal sperm smear in 1% eosin stain was analyzed for SHAs. In DLT, male mice were allowed to mate with females after 1, 3 and 6 weeks of end of pesticide treatment. The uterine contents of the sacrificed females were observed for live and dead implants. The analysis of test and control groups data was done by one way ANOVA at p < 0.05. Results: Exposure of all dose levels of imidacloprid (22, 11 and 5.5 mg/kg/day) for seven days did not induce significant SHAs while they induced significant SHAs compared with the control group following exposure for 14 and 28 days. The analysis of uterine content revealed a significant increase in the number of dead implants/female compared with the vehicle control in only those females which were mated with male mice after six weeks of treatment of highest dose level of imidacloprid. The dominant lethal mutations were observed only at spermatogonial stage. Conclusions: Long-term exposure of pesticide generated SHAs even at lowest dose level (5.5 mg/kg/day for 14 days) and mutagenic effects at spermatogonial stage at highest dose level (22 mg/kg/day for 28 days).

RETRACTED: Chromosome aberrations, micronucleus and sperm head abnormalities in mice treated with Delvocid, a food preservative

Natamycin [corrected] is used as preservative in foods. The genotoxic effects of the food preservative natamycin [corrected] were evaluated using chromosome aberrations and micronucleus test in bone marrow cells and sperm head abnormality assays in mice. Blood samples were taken from mice and levels of total testosterone in serum were also determined. Natamycin [corrected] was intraperitoneally (ip) injected at 200, 400 and 800 mg/kg. Natamycin [corrected] did not induce chromosome aberrations but significantly increased the number of micronucleated polychromatic erythrocytes in bone marrow and sperm head abnormalities at all concentrations and treatment periods. It also decreased MI at all concentrations for 6, 12 and 24h treatment periods. Natamycin [corrected] decreased PCE/NCE ratio at all concentrations for 48h in female mice, for 24 and 48h treatment periods in male mice. At the 800 mg/kg concentration, natamycin [corrected] decreased PCE/NCE ratio for 24 and 72h in female mice. A dose dependent increase was observed in the percentage of sperm head abnormalities. The levels of serum testosterone decreased dose-dependently. The obtained results indicate that natamycin [corrected] is not clastogenic, but it is aneugenic in mice bone marrow and it is a potential germ cell mutagen in sperm cells.

An In Vivo Assay of the Mutagenic Potential of Praziquantel (PZQ) Using Sperm Head Abnormality Test

The mutagenic potential of praziquantel (PZQ) 2-(Cyclohexylcarbonyl) - 1, 2, 3, 6, 7, 11b-hexahydro- 4H-pyrazino-(2,1-a) isoquinolin-4-one, an antihelmintic was evaluated in vivo using sperm head abnormality assay of about 15 weeks-old isogenic mice. For 3, 5, 7 weeks of drug exposure period, three different dose levels 0.02; 0.04; and 0.08 (mg/gbwt) of PZQ (the calculated equivalence of human therapeutic dose-HTD) were intraperitoneally administered. Observations revealed that PZQ had no significant effect on the body weights of the tested animals. Seven types of sperm head morphological abnormalities in varying degrees were observed. Sperm head abnormality was 5.14%, 5.13% and 5.03% in the control mice and 5.62%, 6.56% and 6.72% in the experimental models for the 3, 5 and 7 weeks exposure period respectively. The abnormality was lower in the control as compared to the experimental mice and this represents a difference of 0.48% (3 weeks); 1.43% (5 weeks) and 1.69% (7 weeks). The difference in the abnormal sperm heads between the control and experimental model is statistically insignificant (P<0.05). The induction was slightly dose-dependent with 0.08mg which was the highest dose used inducing the highest number of sperm head abnormalities for each exposure period. PZQ is probably not mutagenic because the values of aberrant sperm heads that were present in the lower dose levels are close to those of the control. PZQ, therefore, may be a mutagenically safe drug for mass therapy and the control of schistosomiasis in Africa.

Mutagenic profiles of carbazole in the male germ cells of Swiss albino mice

Mutagenic effect of carbazole was evaluated by employing dominant lethal mutation and sperm head abnormality assays in male Swiss albino mice. For the dominant lethal mutation assay, adult male mice were treated for five consecutive days either with 30 or 60 mg/kg body weight (b.w.) of carbazole by single intraperitoneal (i.p.) injection. For the sperm head abnormality assay mice were treated with 50, 100, 150, 200 and 300 mg/kg b.w as a single i.p. injection. Treatment of adult male mice with carbazole resulted in induction of dominant lethal mutation and abnormal sperm heads. The results show that carbazole is mutagenic in male germ cells of mice.

Germ cell mutagenicity of phthalic acid in mice

Mutagenicity of phthalic acid was evaluated by employing dominant lethal mutation and sperm head abnormality assays in male Swiss albino mice. For the dominant lethal mutation assay, adult male mice received a single intraperitoneal (i.p.) injection of either 40 mg or 80 mg/kg b.w. of phthalic acid for 5 consecutive days. For the sperm head abnormality assay, the mice were treated with 50, 100, 150, 200 and 300 mg/kg b.w. as a single i.p. injection. Treatment of adult male mice with phthalic acid resulted in induction of dominant lethal mutations and abnormal sperm heads. The results obtained indicate that phthalic acid is a germ cell mutagen.

Testicular toxicity and mutagenicity of steroidal and non-steroidal estrogens in the male mouse

The mutagenicity and toxicity of diethylstilbestrol (DES), 17β-estradiol and zeranol on the male mouse germ cells were investigated with meiotic micronucleus assays in vivo and in vitro, sperm-head abnormality test and morphometry. Further, the developmental effects of DES on the testicular morphology were explored. Micronucleus induction was observed at 10 −7 M concentration of DES and 17β-estradiol in vitro, but other treatments yielded negative results. The micronucleus assay in vivo revealed a small number of micronuclei in early haploid spermatids 17 days after a single subcutaneous injection of DES 50 mg/kg, whereas estradiol and zeranol gave negative results. The sperm-head abnormality rates were significantly elevated 5 weeks after treatments with high doses of DES, 17β-estradiol and zeranol, and testicular morphometry revealed transient changes in the volume densities of testicular tissue components. Prenatal and neonatal estrogen administration resulted in permanent alterations in seminiferous epithelium and dilatation of the rete testis, but did not affect micronucleus or sperm-head abnormality rates. The mutagenicity and toxicity of hormones in the mouse testis paralleled the hormonal activity of these compounds. Early estrogenization was the most sensitive toxicity test, followed by in vitro meiotic micronucleus induction, whereas the sperm-head abnormality assay and morphological analysis did not reveal subtle changes.

Evaluation of the genotoxic potential of six triorganotin compounds by the mouse micronucleus and spermhead abnormality tests

AbstractSix triorganotin compounds—Ph3SnOH, BuPh2SnOH, (p‐CIC6H4)Ph2SnOH, (cyclo‐C5H9)Ph2SnOH, Ph3SnO2CCH2CS2NMe2 and Bu3SnOSO2Et—were tested for their mutagenic potential in somatic and germinal cells in ICR mice by using the micronucleus and spermhead abnormality assays, respectively. In somatic cells, the compunds significantly induced chromosomal disorders at half their respective estimated LD50 (i.p.) values (5.00–6.25 mg kg−1 body wt), and one compound, Ph3SnOH, even at 1/20 of its estimated LD50 value. In germinal cells, five compounds significantly induced chromosomal disorders at 1/64 of their respective estimated LD50 (i.p.) values, whereas one, (p‐CIC6H4)Ph2SnOH, caused such disorders only at 1/8 of its estimated LD50 value.

Activity of 1,1,1- and 1,1,3-trichloroacetones in a chromosomal aberration assay in CHO cells and the micronucleus and spermhead abnormality assays in mice

1,1,1- and 1,1,3-trichloroacetones (TCA) result from the disinfection of municipal water supplies with chlorine, and are direct-acting mutagens in the Ames/Salmonella assay. The objective of this study was to further investigate the genotoxicity of these compounds in mammalian cells using an in vitro chromosomal aberration assay in Chinese hamster ovary (CHO) cells and the micronucleus and spermhead abnormality assays in mice. Both compounds induced significant increases in structural chromosomal aberrations in CHO cells in the presence and in the absence of rat S9 metabolic activation (MA). 1,1,3-TCA was more cytotoxic to CHO cells but 1,1,1-TCA resulted in a higher proportion of cells with aberrations. The clastogenic activities of both compounds were reduced in assays conducted with MA. Neither compound resulted in the induction of a significant increase in micronucleated polychromatic erythrocytes from bone marrow of Swiss-Webster mice when administered by oral gavage; nor were effects seen on the incidence of sperm with head-shape abnormalities, testis weight, or epididymal sperm concentration in B6C3F 1 mice 21 or 35 days after treatment. These data indicate that the drinking water contaminants 1,1,1- and 1,1,3-TCA are clastogenic in vitro, but are not clastogenic to bone marrow cells in vivo, and do not adversely affect several indicators of testicular function in mice.

Effect of primidone in somatic and germ cells of mice

Primidone, an anti-convulsant drug, was tested in mice for mutagenicity in somatic cells by the micronucleus test and in germ cells by the sperm-head abnormality assay. Mice were treated orally with the drug at doses of 4.37, 8.75 and 13.11 mg/mouse. The results indicate that the drug is capable of inducing mutations both in somatic and germ cells of mice.

Evaluation of chemicals used for drinking water disinfection for production of chromosomal damage and sperm-head abnormalities in mice.

Chemical oxidants are commonly added during water treatment for disinfection purposes. These chemicals have not been tested previously for their ability to induce genetic damage in vivo. Chlorine (hypochlorite and hypochlorous acid), monochloramine, chlorine dioxide, sodium chlorite, and sodium chlorate were evaluated for induction of chromosomal aberrations and micronuclei in bone marrow of CD-1 mice, and for induction of sperm-head abnormalities in B6C3F1 mice. Oral administration of chlorine at pH 8.5 (where hypochlorite predominates) at dose levels equivalent to approximately 4 and 8 mg/kg/day induced significant increases in the level of sperm-head abnormalities. There was no evidence of other effects with any of the disinfectants. Halogenated acetonitriles, which have previously been shown to form in the stomach following oral dosing of sodium hypochlorite to rats, were also tested in the sperm-head abnormality assay but gave no indication of an effect.

Evaluation of genotoxic potential of vincristine from multiple parameters

The genotoxic potential of vincristine is assessed by in vivo and in vitro cytogenetic assays on two test systems using multiple parameters. In in vivo experiments it was administered intraperitoneally to Swiss albino mice at doses computed on a surface area basis to lie within the therapeutic range. The parameters employed include screening of chromosome mutational activity in bone marrow and meiotic cells for periods covering one spermatogenetic cycle, in vivo SCE analysis and sperm head abnormality assay. In vitro studies included conventional scoring for chromosome mutations from human leucocyte cultures following administration of the drug at doses within the therapeutic range on the basis of blood volume. Strict controls were employed in both test systems. The data were treated by appropriate statistical tests to evaluate their validity. The results of in vivo experiments on somatic as well as meiotic test systems reveal a lack of clastogenic action of the compound. Both sexes respond in an identical fashion regarding this activity. The SCE analysis supports the in vivo nonclastogenic action of vincristine. Total univalents significantly seen with the highest dose after 1 and 2 weeks must be considered as being important as they can lead to aneuploidy. Evidence is adduced from sperm head abnormality assay that the drug induces point mutations. The results from in vitro test system indicate a positive chromosome mutational property. This is substantiated by results of in vitro SCE analysis. A comparison of data from in vivo and in vitro test systems suggests that a metabolic conversion of the compound in the former might render it innocuous for expressing the chromosome mutational activity. They also point out that vincristine evokes a differential clastogenic response in the two test systems despite bringing about an anaphase arrest commonly in both by acting on cytokinesis.

GOSSYPOL — AN EFFECTIVE MALE CONTRACEPTIVE WAS NOT MUTAGENIC IN SPERM HEAD ABNORMALITY ASSAY IN MICE

Gossypol acetic acid, a potential male anti-fertility compound, was investigated for its mutagenic effects in the mouse sperm head abnormality assay. Compared with the controls, the chemical at 20 and 40 mg/kg doses did not increase detectable sperm head abnormalities in mice.

Sperm head abnormality test

Benzyl chloride, 4-chloromethylbiphenyl and 4-hydroxymethylbiphenyl either subcutaneously or intraperitoneally at up to lethal doses did not induce a positive response in mice in the sperm head abnormality assay in mice.