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The number of animals, which died during this interval, was expressed as a percentile, and the LD50 determined by probit test using a death percent versus doses' log [ 40].
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The number of deaths was expressed as a percentile and the LD50 was determined by probit a test using the death percentage versus the log dose [ 12].
LC20, LC50, and LC80 values were obtained by Probit analysis using SAS software (SAS Institute 2004).
CC50 was determined by probit analysis using the Pharm PCS (Pharmacologic Calculation System) statistical package (Springer-Verlag, USA).
The LoD was inferred by probit analysis using the IBM SPSS Statistics 20 software.
LD50 values and 95% confidence intervals were calculated by probit analysis using SPSS v.13 (SPSS Inc, Chicago, IL).
LC50 values were calculated by Probit analysis using the EPA Probit Analysis program, version 1.5 (US Environmental Protection Agency, Cincinnati, OH, USA).
IC10 and IC50 values were determined by probit analysis using a macro program for Microsoft Excel written by J. Greve (Fraunhofer-Institut für Umweltchemie und Ökotoxikologie, Schmallenberg, Germany).
Analysis of the in vitro tests using Probit (SAS program) allowed the calculation of lethal concentrations (LCs).
A Kruskal-Wallis test, P <0.05 was used to determine significant differences [ 22] and a PROBIT test was performed with POLO PLUS [ 23] to determine the LC50, LC90 and LC99 of the extracts that showed in vitro fascioliscide efficacy.
Dose response data were analyzed by probit method [ 12] using GraphPad Prism 4 software.
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