At 36 hours http://www.selleckchem.com/products/epz-5676.html after administration of treatments, peripheral blood samples were collected by perforating the caudal vein, and drops were placed at the prestained slides with acridine orange as described by Hayashi and Sofuni, 1994 [15]. The micronucleated cells were scored under a fluorescence microscope. 1000 peripheral reticulocytes per mouse were analyzed, and the frequencies of micronucleated peripheral reticulocytes were scored in three slides per animal [16].2.6. Statistical AnalysesLD50 value was determined through Probit analysis [17]. One-way ANOVA multicomparisons tests were used to identify any significant difference among genotoxicity tests between animal groups, and Fisher’s least significant difference (LSD) was used to compare significant differences between groups. A P value < 0.

05 was considered statistically significant. All data was analyzed using the software Statgraphics Version XVI.I.3. Results3.1. Acute ToxicityThe P. americana seed extract administrated at doses of 500, 1000, and 2000mg/Kg showed a mortality of 20, 60, and 80%, respectively. The animal groups treated with 125 and 250mg/Kg, as well as the control group, showed no mortality (Figure 1). The LD50 calculated for the avocado seed extract was 1200.75mg/kg. According to these results, a concentration of 250mg/Kg was determined for genotoxicity testing.Figure 1Percentage of mortality for determining the acute toxicity of the P. americana seed extract.3.2. Genotoxicity TestThe animal groups administered with 250mg/Kg of P.

americana seed extract and the negative control group showed a low amount of micronucleated cells, while the positive control administered with colchicine showed clear evidence of harm. There is no statistically significant difference between the group administered with the extract of P. americana seed and control group; however, there is a significant difference between both these groups in regard to the positive control (Table 1).Table 1Micronucleated peripheral reticulocytes (MNRET) formed by P. americana seed extract.4. DiscussionThe avocado (P. americana) is consumed by humans because of its organoleptic characteristics; furthermore, the pulp contains up to 33% oil, rich in monounsaturated fatty acids [3]. The avocado seed is discarded in the majority of countries, although in some countries such as Niger, it is consumed [18, 19]. This waste may represent an ecological or human contaminant.In a study previously reported on acute and subacute toxicity of a P. americana aqueous extract, it was not possible to estimate the LD50 value with the doses GSK-3 tested (up to 10g/kg). Also, in repeated doses, toxicity tests during 28 days showed no affectations in hematological and biochemical parameters.