Acute Toxicity of Keladi Tikus (Typhonium flagelliforme (Lodd.) Blume) Ethanol Extract on Zebrafish (Danio rerio) Embryo In Vivo

Author(s):  
Fajar Fakri ◽  
Loly Subhiaty Idrus ◽  
Maria Alexandra Iskandar ◽  
Indra Wibowo ◽  
I Ketut Adnyana

Keladi tikus (Typhonium flagelliforme (Lodd.) Blume) is an Indonesian medicinal plant that has various pharmacological properties. Zebrafish (Danio rerio) has been proposed as a model that can bridge the gap between cell assays and rodent assays. Evaluation of the toxic effects of natural products using the Zebrafish model can be assessed starting from the blastula stage of embryonic development. This study aims to investigate the potential acute toxicity effect of keladi tikus-ethanol extract (KTEE) using zebrafish embryos. A static non-replacement regime for acute toxicity testing was used. Wild-type zebrafish embryos were exposed to various concentrations of KTEE (50, 100, 200, 400, 800, 1600 µg/mL) starting at 6 hours post-fertilization (hpf) until 96 hpf. The results showed that the survival rate of zebrafish embryos decreased as the concentration of the test extract increased. The LC50 values of KTEE were 494.553 µg/mL at 96 hpf and 555.787 µg/mL at 72 hpf. Embryotoxicity effect of KTEE includes hatching delays and decreased heartrate on zebrafish embryos, especially at high concentrations. KTEE also caused abnormalities in embryo morphology, including pericardial edema, jaw and tail deformity.

Author(s):  
Gustini Syahbirin ◽  
Nurfadilawati Mumuh ◽  
Kusdiantoro Mohamad

Objective: This study was aimed at determining the levels of curcuminoids and analyzing the toxicity of ethanol extracts of Javanese ginger.Methods: Curcuminoid levels were determined using high-performance liquid chromatography, while the toxicity tests were done on larva of brine shrimp (Artemia salina) by using a brine shrimp lethality test (BSLT) method and embryos of zebrafish (Danio rerio) using a zebrafish embryo acute toxicity (ZFET) method.Results: The level curcuminoid of ethanol extracts was 10.5% dry wt., consisting of curcumin at the highest percentage (68.06%) followed bydesmethoxycurcumin (24.6%) and bisdemethoxycurcumin (1.41%). In BSLT method, the lethal concentration 50% values (LC) value of our ethanol extract was 238 ppm, whereas in ZFET method, the LC value at 96 hours after fertilization was 80 ppm. The ethanol extract of ginger caused major malformations of the pericardial edema of zebrafish embryos at a concentration of 100 ppm. 50Conclusion: The ethanol extract of Curcuma xanthorrhiza from Bogor contained curcuminoids consisting of curcumin, desmethoxycurcumin, and bisdemethoxycurcumin, with acute toxicity, caused major malformations on the pericardial edema in zebrafish embryos.Keywords: Curcuma xanthorrhiza, Curcuminoid, Toxicity, Zebrafish embryo acute toxicity.50


2016 ◽  
Vol 40 (8) ◽  
pp. 6599-6603 ◽  
Author(s):  
Prusothman Yoganantharajah ◽  
Daniel J. Eyckens ◽  
Jessie L. Pedrina ◽  
Luke C. Henderson ◽  
Yann Gibert

The in vivo toxicity of several solvate ionic liquids have been assessed using a zebrafish model.


Molecules ◽  
2018 ◽  
Vol 24 (1) ◽  
pp. 8 ◽  
Author(s):  
Mayra Antúnez-Mojica ◽  
Andrés Rojas-Sepúlveda ◽  
Mario Mendieta-Serrano ◽  
Leticia Gonzalez-Maya ◽  
Silvia Marquina ◽  
...  

By using a zebrafish embryo model to guide the chromatographic fractionation of antimitotic secondary metabolites, seven podophyllotoxin-type lignans were isolated from a hydroalcoholic extract obtained from the steam bark of Bursera fagaroides. The compounds were identified as podophyllotoxin (1), β-peltatin-A-methylether (2), 5′-desmethoxy-β-peltatin-A-methylether (3), desmethoxy-yatein (4), desoxypodophyllotoxin (5), burseranin (6), and acetyl podophyllotoxin (7). The biological effects on mitosis, cell migration, and microtubule cytoskeleton remodeling of lignans 1–7 were further evaluated in zebrafish embryos by whole-mount immunolocalization of the mitotic marker phospho-histone H3 and by a tubulin antibody. We found that lignans 1, 2, 4, and 7 induced mitotic arrest, delayed cell migration, and disrupted the microtubule cytoskeleton in zebrafish embryos. Furthermore, microtubule cytoskeleton destabilization was observed also in PC3 cells, except for 7. Therefore, these results demonstrate that the cytotoxic activity of 1, 2, and 4 is mediated by their microtubule-destabilizing activity. In general, the in vivo and in vitro models here used displayed equivalent mitotic effects, which allows us to conclude that the zebrafish model can be a fast and cheap in vivo model that can be used to identify antimitotic natural products through bioassay-guided fractionation.


2010 ◽  
Vol 42 (2) ◽  
pp. 300-309 ◽  
Author(s):  
Jordan T. Shin ◽  
Eugene V. Pomerantsev ◽  
John D. Mably ◽  
Calum A. MacRae

Phenotype-driven screens in larval zebrafish have transformed our understanding of the molecular basis of cardiovascular development. Screens to define the genetic determinants of physiological phenotypes have been slow to materialize as a result of the limited number of validated in vivo assays with relevant dynamic range. To enable rigorous assessment of cardiovascular physiology in living zebrafish embryos, we developed a suite of software tools for the analysis of high-speed video microscopic images and validated these, using established cardiomyopathy models in zebrafish as well as modulation of the nitric oxide (NO) pathway. Quantitative analysis in wild-type fish exposed to NO or in a zebrafish model of dilated cardiomyopathy demonstrated that these tools detect significant differences in ventricular chamber size, ventricular performance, and aortic flow velocity in zebrafish embryos across a large dynamic range. These methods also were able to establish the effects of the classic pharmacological agents isoproterenol, ouabain, and verapamil on cardiovascular physiology in zebrafish embryos. Sequence conservation between zebrafish and mammals of key amino acids in the pharmacological targets of these agents correlated with the functional orthology of the physiological response. These data provide evidence that the quantitative evaluation of subtle physiological differences in zebrafish can be accomplished at a resolution and with a dynamic range comparable to those achieved in mammals and provides a mechanism for genetic and small-molecule dissection of functional pathways in this model organism.


Author(s):  
Noor Izati Abd Aziz ◽  
Vikneswari Perumal ◽  
Suganya Murugesu ◽  
Qamar Uddin Ahmed ◽  
Bisha Fathamah Uzir ◽  
...  

 The use of zebrafish vertebrate model in vivo analysis of the drug toxicity and efficacy, chemical toxicity, and safety is increasing in recent researches. Momordica charantia Linn (Cucurbitaceae) has been traditionally claimed for its many protective roles. However, the development of toxicity effect may cause morphological abnormalities by using an embryo of zebrafish (Danio Rerio) is unknown. Hence, this study was designed to determine the toxicity and teratogenic effect of hydroethanolic extract of M. charantia fruit using Zebrafish (Danio Rerio) embryos. The crude extract was prepared from the fruit of M. charantia using 80% hydroethanolic solvent. The zebrafish embryos were exposed to serial dilution of crude extract. The active constituent was analyzed using gas chromatography coupled with mass spectrophotometry (GC-MS) Momordica charantia Linn (Cucurbitaceae) has been widely commercialized based on traditional usage as an antidiabetic product. The current study has shown the toxic effects of the M.  charantia fruit extract on the developing zebrafish embryos, and the median lethal concentration (LC50) was calculated to be 725.90 mg/L at 48 hpt. The observed effects are dependent on the time of exposure and concentrations of the extract. At higher concentration, the extract causes some morphological defects such as less pigmentation, dented tail, spinal curvature, oedema, reduced hatchability, and growth retardation, that indicates the presence of toxicant(s). Based on the GC-MS profiling, some of the compounds identified in the hydroethanolic extract, such as propanedioic acid and glutamine, may have caused the teratogenic effects to the embryos. Further research on the M. charantia fruit's metabolites should be carried out prior to any nutraceutical or pharmaceutical application.


Molecules ◽  
2020 ◽  
Vol 25 (19) ◽  
pp. 4482
Author(s):  
Bryan Veeren ◽  
Batoul Ghaddar ◽  
Matthieu Bringart ◽  
Shaymaa Khazaal ◽  
Marie-Paule Gonthier ◽  
...  

Antirhea borbonica (A. borbonica) is an endemic plant from the Mascarene archipelago in the Indian Ocean commonly used in traditional medicine for its health benefits. This study aims (1) at exploring polyphenols profiles from two types of extracts—aqueous (herbal infusion) and acetonic (polyphenol rich) extracts from A. borbonica leaves—and (2) at evaluating their potential toxicity in vivo for the first time. We first demonstrated that, whatever type of extraction is used, both extracts displayed significant antioxidant properties and acid phenolic and flavonoid contents. By using selective liquid chromatography–tandem mass spectrometry, we performed polyphenol identification and quantification. Among the 19 identified polyphenols, we reported that the main ones were caffeic acid derivatives and quercetin-3-O-rutinoside. Then, we performed a Fish Embryo Acute Toxicity test to assess the toxicity of both extracts following the Organisation for Economic Cooperation and Development (OECD) guidelines. In both zebrafish embryos and larvae, the polyphenols-rich extract obtained by acetonic extraction followed by evaporation and resuspension in water exhibits a higher toxic effect with a median lethal concentration (LC50: 5.6 g/L) compared to the aqueous extract (LC50: 20.3 g/L). Our data also reveal that at non-lethal concentrations of 2.3 and 7.2 g/L for the polyphenol-rich extract and herbal infusion, respectively, morphological malformations such as spinal curvature, pericardial edema, and developmental delay may occur. In conclusion, our study strongly suggests that the evaluation of the toxicity of medicinal plants should be systematically carried out and considered when studying therapeutic effects on living organisms.


2010 ◽  
Vol 18 (3) ◽  
pp. 1038-1044 ◽  
Author(s):  
Livio Racané ◽  
Marijeta Kralj ◽  
Lidija Šuman ◽  
Ranko Stojković ◽  
Vesna Tralić-Kulenović ◽  
...  

2020 ◽  
Vol 3 (2) ◽  
pp. 1
Author(s):  
Romel Ciptoadi Wijaya

Preface :Patchouli Oil requires toxicity testing for safety before we can use it widely. It causes side effects such as nausea, vomiting and loss of appetite in some people. Determination of lethal concentration 50% (LC50) in the early stages of zebrafish embryos development will provide an easier, faster and precise prediction of toxicity. At a certain dose, it can cause impairment and death toward organisms. Therefore, the aim of this study was to determine LC50 of Patchouli Oil in carboximethyl cellulose emulsifier towards zebrafish embryo (Danio rerio).Method :Laboratory experimental study was using zebrafish embryos at 2 hours post fertilization. The total of 160 embryos were used and divided into 8 groups, i.e.; Negative control (KN) was given the embryonic fluid, positive control 1 was given 5000 ppm of patchouli oil (KP1), positive control was given 5000 ppm of CMC (KP2), and 5 treatment groups, i.e.; concentration of 10 ppm (P1), 30 ppm (P2), 60 ppm (P3), 90 ppm (P4) and 120 ppm (P5) of Patchouli oil which was emulsified in CMC. The study was conducted with 3 times repetition. Data collection was done by calculating total of embryo’s deaths for each treatment at 24-72 hours of exposure. Data were analyzed using Regression Probit Analysis.Results :Mortality in the group of KN was 1.6%, KP1 was 98.3% and KP2 was 0%. Meanwhile in the treatment group of P1, P2, P3, P4 and P5 respectively were 0%, 5%, 10%, 25%, and 50%.Conclusion :Based on these results, Lethal Concentration 50% of Patchouli Oil in CMC emulsifier towards zebrafish embryo is 120 ppm.


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