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Phenylhydrazine hydrochloride induced dose dependent embryo cytotoxicity in zebrafish


Raskin Rajagopal*, Meenakshi Balasubramanian & Shantaraman Kalyanaraman



Tirunelveli Medical College, Tirunelveli, TamilNadu 627011, India; Phone: +91 9962666658



Raskin Rajagopal - E-mail:; *Corresponding author


Article Type

Research Article



Received March 14, 2019; Accepted March 23, 2019; Published April 15, 2019



Phenylhydrazine hydrochloride (PHZ) is a chemical compound. PHZ and its derivatives were used firstly as antipyretics, treatment of blood disorders such as polycythaemia Vera. For many years phenylhydrazine was used for experimental induction of anaemia in animal models. However, this compound is reported to cause damage to red blood cells, potentially resulting in anaemia and consequential secondary involvement of other tissues, such as the spleen and liver. Recent studies suggest that PHZ cause genotoxicity in mice models. The aim of our study is to study the effect of PHZ in embryonic and larval stage of zebra fish model. Zebra fish embryos and larvae were used in this study. Working concentration prepared from 0.05 gm of PHZ stock solution. The embryos and larvae were exposed to different concentrations of PHZ (0.1, 0.3, 0.5, 0.7 0.9, 1.0, 3.0, 5.0, 7.0 9.0 and 10.0 μg/mL) and (0.1, 0.3, 0.5, 0.7, 0.9 and 1.0 μg/mL) respectively. Survival rate, mortality rate, hatching rate and phenotypic anomalies were studied in developing embryos. Heart rate and apoptosis were evaluated to assess the PHZ toxicity in larval stage of Zebra fish. Statistical analysis was performed by Pearson correlation and P values < 0.05 were considered statistically significant. The LC50of PHZ in embryo and larvae was found to be 0.7μg/mL. PHZ treated embryos showed that survival rate was decreased during 72hpf. In the case of mortality, 0.7 μg/mL and above concentration mortality rate was significantly increased between 48 and 72 hpf and the none embryos survived after 72 hpf. We observed delayed hatching rate in treated embryos when compared to control embryos. 0.5 μg/mL treated larvae showed significantly (p<0.05) decreased heart rate 20% at 96 hrs. Phenotype anomalies such as enlarged yolk sac, yolk sac split, pericardial edema, notochord anomaly appeared at higher concentration of PHZ treated embryos. Acridine orange fluorescence staining revealed that high apoptotic cells were detected at caudal fin region of larvae on day 3 at a concentration of 0.7μg/mL treated group. Our study suggests that PHZ causes multiple phenotypic abnormalities and toxicity on zebrafish embryos and larvae with respect of dose and time dependent manner.



Phenylhydrazine hydrochloride, cytotoxicity, zebrafish



Rajagopal et al. Bioinformation 15(4): 255-260 (2019)


Edited by

P Kangueane






Biomedical Informatics



This is an Open Access article which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. This is distributed under the terms of the Creative Commons Attribution License.