Journal of ethnopharmacology · 2025
Toxicity assessment of Eucomis autumnalis and Merwilla plumbea using the in vivo zebrafish embryotoxicity test (ZET).
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Kanama Kevine Sowesa, Vermaak Ilze, Sandasi Maxleene, Chen Weiyang, Ncube Efficient, Viljoen Alvaro
Аннотация
ETHNOPHARMACOLOGICAL RELEVANCE: Eucomis autumnalis (Mill.) Chitt. and Merwilla plumbea (Lindl.) Speta are popularly used in southern African healing practices to treat a wide range of diseases such as gastrointestinal, respiratory and skin conditions, as well as spiritual protectants against witchcraft. The pharmacological properties of the two species are well documented; however, studies that investigate the safety profiles using in vivo models are limited.
AIM OF THE STUDY: This study evaluated the toxicity profiles of the two plant species independently using the in vivo zebrafish embryotoxicity test (ZET).
MATERIALS AND METHODS: The whole plant of E. autumnalis and the bulb of M. plumbea were extracted with methanol using ultrasonic-assisted extraction. Zebrafish embryos at 6 hours post-fertilisation (hpf) were placed in microtitre plates and treated with various concentrations of methanol extracts of the two plant species. The plates were incubated at 28.5 °C and embryos observed at 24, 48, 72, and 96 hpf on an inverted microscope mounted with a camera and a high-speed digital charge-coupled device (CCD). Acute toxicity (hatchability, survival rate and LC50 values), embryo activity (burst activity and burst count/min), cardiotoxicity (heart rate and blood flow), and morphological defects were monitored and activity videos analysed using DanioScope V1 software.
RESULTS: The ZET test demonstrated that both E. autumnalis and M. plumbea methanol extracts are embryotoxic with LC50 values of 9.7 μg/mL and 7.65 μg/mL, respectively. At higher concentrations, E. autumnalis (10-12 μg/mL) and M. plumbea (7 μg/mL - 10.5 μg/mL) increased burst activity and induced abnormalities in the yolk sac and swim bladder development. Eucomis autumnalis did not cause cardiotoxic effects at all the tested concentrations; however, M. plumbea treatment resulted in decreased heart rate at concentrations >9.5 μg/mL, indicating cardiotoxicity.
CONCLUSION: This study emphasises the importance of toxicological-related research on medicinal plants to ensure documentation of possible toxic effects.
AIM OF THE STUDY: This study evaluated the toxicity profiles of the two plant species independently using the in vivo zebrafish embryotoxicity test (ZET).
MATERIALS AND METHODS: The whole plant of E. autumnalis and the bulb of M. plumbea were extracted with methanol using ultrasonic-assisted extraction. Zebrafish embryos at 6 hours post-fertilisation (hpf) were placed in microtitre plates and treated with various concentrations of methanol extracts of the two plant species. The plates were incubated at 28.5 °C and embryos observed at 24, 48, 72, and 96 hpf on an inverted microscope mounted with a camera and a high-speed digital charge-coupled device (CCD). Acute toxicity (hatchability, survival rate and LC50 values), embryo activity (burst activity and burst count/min), cardiotoxicity (heart rate and blood flow), and morphological defects were monitored and activity videos analysed using DanioScope V1 software.
RESULTS: The ZET test demonstrated that both E. autumnalis and M. plumbea methanol extracts are embryotoxic with LC50 values of 9.7 μg/mL and 7.65 μg/mL, respectively. At higher concentrations, E. autumnalis (10-12 μg/mL) and M. plumbea (7 μg/mL - 10.5 μg/mL) increased burst activity and induced abnormalities in the yolk sac and swim bladder development. Eucomis autumnalis did not cause cardiotoxic effects at all the tested concentrations; however, M. plumbea treatment resulted in decreased heart rate at concentrations >9.5 μg/mL, indicating cardiotoxicity.
CONCLUSION: This study emphasises the importance of toxicological-related research on medicinal plants to ensure documentation of possible toxic effects.