Allium Cepa Test Research Articles

Evaluation of Cytotoxic and Genotoxic Activities of Croton zambesicus Muell Arg. Leaf Extract

Croton zambesicus Muell Arg. (Euphorbiaceace) a medicinal plant used traditionally in the treatment of diseases was evaluated for genotoxic and cytotoxic effects using Allium cepa (onion) test. The effect of the leaf extract on the root meristematic cells of Allium cepa was assessed macroscopically and microscopically using onion bulbs exposed to 2.5, 5 and 10 mg/mL concentrations of the extract. Tap water and methotrexate (0.1 mg/mL) were used as a negative and positive control respectively. The extract significantly (P < 0.05) caused concentration-dependent inhibition of the onion root growth. The extract exerted cytotoxic effects on cell division in A. cepa. The extract-induced chromosomal aberrations in A. cepa root tip cells and root growth inhibition were significant (P < 0.05) when compared with the control group. Treatment with the extract resulted in cell death, ghost cells, cells membrane damage, and binucleated cells. These results suggest that the leaf extract of Croton zambesicus possess cytotoxic and genotoxic effects on A. cepa.

Evaluation of the Performance of Microalgae and Constructed Wetlands for the Reduction of Toxicity Organic Load and Pharmaceuticals from Wastewaters

The present study aimed to evaluate the performance of a novel integrated system composed of microalgae and constructed wetlands (CW). The microalgae production tank was designed as a raceway type, and microalgae of the genus Chlorella were inoculated, while the CW was established as the first stage of the French model and vegetated with the macrophytes Chrysopogon zizanioides, Typha domingensis, and Dracaena trifaciata. The hydraulic retention time in each unit was 7 days, totaling 14 days of treatment. Good reductions in dissolved organic carbon (67.2%) and Total N (68.6%) were achieved after the treatment. The treatment also completely eliminated acute ecotoxicity against Daphnia magna and genotoxicity in the Allium cepa test assay. Liquid chromatographic analysis revealed the presence of 9 pharmaceuticals in the raw wastewaters, whereas only residual dipyrone was detected after the CW unit. Thus, the novel system investigated in our study, which combines microalgae and CW using 3 different species of macrophytes, proved to be a promising alternative for treating urban wastewaters, as it effectively reduced organic load, nutrient content, toxicity, and removed emerging contaminants such as pharmaceuticals. Future recommendations include investigating the primary removal mechanisms of the pharmaceuticals and enhancing the separation process of the microalgae.

Biomonitoring of the Paraopeba river: Cytotoxic, genotoxic and metal concentration analysis three years after the Brumadinho dam rupture - Minas Gerais, Brazil.

The rupture of Vale S.A. mining tailings dam in Brumadinho, Brazil, in January 2019 had significant environmental impacts on the Paraopeba River basin. Additionally, severe floods in early 2022 contributed to the transport of particles in the river. This study aimed to evaluate the cytotoxic and genotoxic potential of Paraopeba River water. Thus, the Allium cepa test system was applied, along with physicochemical analyses, flow cytometry, and metal concentration, comparing the results between the rainy and dry seasons three years after the dam rupture. The tests were conducted on water samples collected during three periods: January 2022, July 2022, and January 2023, at five points along the river and its tributaries. Allium cepa seeds were exposed to the collected water samples, as well as negative (water) and positive (trifluralin) controls. Cytotoxicity was evaluated using the mitotic index and flow cytometry, and genotoxicity by the chromosomal alterations index. The analysis of metals and physicochemical parameters revealed that most values complied with current regulations. However, there were exceptions, with ammonia levels exceeding the permitted limits at all points in the three collections. High levels of aluminum, iron and nitrite were found at most points, before and after the dam collapse, mainly during the rainy season. This indicates the impact of rainfall on water quality, which increases the transport of contaminating particles, probably resulting from human activities and the high concentration of nitrogen compounds released into the Paraopeba River. The results of the bioassay suggest a relatively low cytotoxic and genotoxic potential of the samples evaluated. However, this study highlights the continuous contamination of the river by unidentified anthropogenic factors, requiring continuous monitoring and analysis to track the evolution of water quality and its environmental effects.

Analysis of water quality, bioindicators, contaminants and their cytogenetic impacts in a Cerrado reservoir

The fragmentation of Brazilian rivers due to the construction of hydroelectric dams has altered aquatic ecosystems, resulting in the accumulation of residues in sediments. This study evaluated the cytotoxicity and genotoxicity of water and sediments from the Cana Brava reservoir, located in the Tocantins-Araguaia basin in the Cerrado biome, using the Allium cepa test. Sampling points were situated in the Cana Brava reservoir near two tributaries, the Bonito River and the Varjão Stream, areas susceptible to contaminant input due to proximity to urban areas. Physicochemical parameters, metals through atomic absorption spectrometry, toxicity, and analyses of bioindicators such as microalgae and macrophytes were assessed. Near the Varjão Stream, algal richness was higher, with the presence of Chara rusbyana, which indicates good water quality. Near the Bonito River, pollution-indicating microalgae such as Euglena sp., Nitzschia sp., and Oscillatoria sp. were identified, along with a prevalence of Salvinia auriculata, favored by nutrient availability. Cytogenetic tests revealed that sediments, especially from the site near the Varjão Stream, caused chromosomal abnormalities and cytogenotoxic effects, with an increase observed after 72 hours. Principal components analysis showed that phosphorus and potassium in the sediment from the site near the Bonito River correlated with cellular alterations at 48 hours, while sediments near the Varjão Stream resulted in a higher frequency of cytogenetic alterations at 72 hours. These results highlight local impacts on water quality and biological diversity, recommending continuous monitoring to mitigate the adverse effects of pollution in the Cana Brava reservoir.

Cytogenetic and photosynthetic responses of plants after exposure to water from a lake environment.

Plants are sensitive to environmental pollutants and are excellent organisms for genetic and physiological testing. Plant-based test systems are often used to study aquatic, aerial, and terrestrial pollution, especially Allium cepa, but studies with Tradescatia pallida specimens have gained prominence due to their sensitivity and applicability. Among the biomarkers, cytogenetic damage and chlorophyll levels are used in stress studies due to their responses to single or combined factors. The aim of this study was to evaluate cytogenetic and photosynthetic responses in T. pallida, and cytogenetic responses in A. cepa exposed to water from three sampling stations in the Juara lagoon (Municipality of Serra, ES, Brazil), collected in two sampling campaigns. The cytotoxic, genotoxic, and mutagenic potentials were analyzed using the T. pallida root tip mitosis assay and the Allium cepa test. Chloroplast pigment levels were measured in T. pallida leaves after chronic exposure to the lagoon water. The cytogenetic tests showed cytogenetic alterations at two sampling stations in at least one sampling campaign, suggesting the presence of potential pollutants, the effects of which were maximized during the rainy season. The study of photosynthetic metabolism in T. pallida showed a relationship between the levels of chloroplast pigments and the amount of nutrients present in the water.

Phytochemical Content and Potential Antioxidants, Antimitotic, Antifungal Effects of Anabasis aretioïdes Coss. & Moq. (Chenopodiaceae)

Anabasis aretioides is an endemic species of the southwestern Algerian region. It is often used as a remedy for its medicinal, antioxidant, antidiabetic, anti-inflammatory, and antimicrobial effects.The aim of this work is to evaluate its biological activities by testing its antioxidant, antiproliferative (Allium cepa test), anti-inflammatory, and antifungal powers. Phytochemical screening of A. aretioides revealed the presence of tannins, free desquinones, terpenoids, fatty acids, saponins, C-heterosides, sterols, and triterpenoid. Furthermore, the in vitro evaluation of the aqueous and methanolic extracts of Anabasis aretioide leaves allowed us to observe the significant DPPH radical scavenging activity of the methanolic extract compared with that of vitamin C (the activity remains significant compared with that of the aqueous extract). The mitotic index (MI) of the methanolic extract revealed that a dose of 1 mg/100 mL had the greatest effect (MI=13.10%), followed by a dose of 2.5 mg/mL (MI=11.22%). In parallel, the analysis of the protective activity against protein denaturation of the extracts revealed that the methanolic extract was relatively superior and very important compared with diclofenac. Additionally, the methanolic extract of Anabasis aretioides against different fungal species revealed significant inhibitory activity against Aspergillus parasiticus, Penicillium glandicola, and Aspergillus at 100 mg/mL. Conclusion, Anabasis aretioides is a plant that may have interesting phytotherapeutic effects.

Toxicological assessment of reactive blue 19 dye aqueous solutions under UV-LED light

Abstract The dye-contaminated industrial effluent causes serious health issues when it gets mixed with underground water without primary treatment. The current project was designed to treat reactive blue-19 dye aqueous solutions in the presence of hydrogen peroxide (H2O2) and titanium dioxide (TiO2) under UV-LED light. The characterization of the photocatalyst was carried out via X-ray diffraction (XRD) and Scanning Electron Microscopy (SEM) for structure, purity, and surface study. The effect of various factors such as pH, TiO2 dose, UV-LED light exposure time, H2O2, and dye concentration, on the degradation rate and cytotoxicity reduction was evaluated and optimized through the Response Surface Methodology (RSM). The maximum degradation of dye solution and chemical oxygen demand (COD) reduction was achieved at 98.81 and 86.22 %, respectively for 50 ppm solution, using UV-LED/H2O2(3 %)/TiO2(6 g/L) hybrid process. The toxicity evaluation through the Allium cepa test demonstrated a 62.40, 65.2, and 56.97 % increase in root length (RL), root count (RC), and mitotic index (MI), respectively, following treatment with the UV-LED/H₂O₂/TiO₂ combined process for 150 min. The hemolytic and brine shrimp tests revealed a reduction in toxicity up to 92.18 and 84.08 %, respectively, after applying the same treatment. Additionally, the Ames test indicated up to 80.94 % reduction in mutagenicity for TA98 and an 84.04 % reduction for TA100 strain when dye samples were treated with UV-LED light in the presence of H₂O₂ and TiO₂ for 150 min. The findings suggested that UV-LED light in conjunction with H2O2 and TiO2 can be a useful tool for the degradation and detoxification of toxic pollutants found in textile wastewater.

Evaluation of antioxidant potential of anthocyanin-rich hydro ethanol extract of Syzygium cumini fruit pulp in various in vitro models and in food supplement enriched with omega-3 fatty acids

Omega-3 fatty acids have immense health benefits but are highly susceptible to oxidation due to high degree of unsaturation in their molecular structure. Therefore, development of omega-3 fatty acids fortified stable functional food is a challenging task. The aim of the study was to evaluate antioxidant potential of anthocyanin-rich hydro ethanol extract of Syzygium cumini fruit pulp extract in various in vitro models and in omega-3 fatty acids enriched food supplement in bulk oil and oil-in-water emulsions model systems along with chemical analysis of the fruit pulp extract and evaluation of its cytotoxic potential, if any. The in vitro antioxidant potential was evaluated by DPPH radical scavenging, ABTS°+ decolorization and Fe2+ ion chelating methods. Antioxidant activity against oxidation of omega-3 fatty acids enriched food supplement in bulk oil and oil-in-water emulsions model systems was evaluated by monitoring primary oxidation, secondary oxidation and total oxidation of the omega-3 enriched oil using AOCS guidelines and iodometric method. Chemical analysis was performed by HR-LCMS analysis technique. Cytotoxic potential was evaluated by Allium cepa test. The test extract exhibited varying degrees of antioxidant activity in various in vitro models as well as against oxidation of omega-3 fatty acids enriched food supplement both in bulk oil and oil-in-water emulsions model systems. Chemical analysis revealed the presence of three major tentatively identified compounds malvidin-3,5-diglucoside, delphinidin-3,5-diglucoside and peonidin-3,5-diglucoside as major components of the extract. Test extract did not show any cytotoxic potential at recommended dosage level with LC50> 1000 µg/ml. The results provide evidence that anthocyanin-rich hydro ethanol extract of Syzygium cumini fruit pulp may serve as a potential source of safe and effective, novel natural antioxidant for retarding oxidation of omega-3 fatty acids enriched food supplement which subsequently may help to develop omega-3 fortified stable functional foods in food industry.

Biological activities of Stryphnodendron adstringens bark decoction

Stryphnodendron adstringens, called ‘barbatimão’, is used in Brazilian folk medicine given its healing, astringent, anti-inflammatory, and antimicrobial properties. However, data on its safe use remain inconclusive due to controversy about the risk posed by its ethnopharmacological intake to human health. Thus, the present study aims to quantify the main classes of compounds found in S. adstringens bark powder and investigate the biological activity of ‘barbatimão’ bark decoction against pathogenic bacterial strains. We also investigated its toxic activity using the Artemia salina test and its cytogenotoxicity using the Allium cepa test. Phenols (24.5 ± 1.5 mg mL-1), tannins (15.08 ± 0.02 mg mL-1), and flavonoids (1.33 ± 0.02 mg mL-1) were quantified in the S. adstringens decoction. This decoction exhibited antibacterial activity against Gram-positive strains and toxicity against A. salina. It was not possible to observe the S. adstringens bark decoction genotoxicity in the A. cepa roots because the cell cycle stopped at the prophase, indicating the decoction’s anti-proliferation effect. S. adstringens is a plant with great herbal potential; therefore, other study models must be used to investigate its potential antibacterial and anti-proliferation applications

Artisanal Gem Mining in Brazil: Evaluation of Oxidative Stress and Genotoxicity Biomarkers.

This study was carried out in the district of Taquaral de Minas, in the municipality of Itinga, located in Jequitinhonha Valley, state of Minas Gerais, which is considered one of the largest yolk-producing regions in Brazil. Miners in gem extraction areas are prone to severe oxidative damage due to their increased exposure to toxic metals, as well as chemical, physical, and biological agents, resulting in diseases such as silicosis. Thus, this work aimed to evaluate occupational exposure in prospectors through biomonitoring techniques using a variety of biomarkers for oxidative stress, genotoxicity, and mutagenicity. Twenty-two miners and seventeen workers who were not occupationally exposed were recruited, totaling thirty-nine participants. The study was approved by the Research Ethics Committee of the Federal University of the Jequitinhonha and Mucuri Valleys. In this study, the levels of total peroxides, catalase activity, and microelements in plasma were evaluated. Additionally, environmental analysis was carried out through the Ames and Allium cepa tests. The results of the lipoperoxidation assessment were significant, with increased frequencies in exposed individuals compared to controls (p < 0.05), as determined by the Mann-Whitney test. Micronutrients in the blood showed lower concentrations in the group exposed to Fe and Se than in individuals not exposed to these elements. The results of the Ames test and Allium cepa test were statistically significant compared to the controls (p < 0.05), as determined by the Mann-Whitney test for genotoxicity and cytotoxicity. Thus, the results of the present study indicate possible environmental contamination and a potential risk to the health of miners, which suggests that further studies are important in the region.

Approach regarding the biosafety evaluation of black and red currant pomace extracts using Allium cepa test

Abstract In the present work, biosafety evaluation of black and red currant pomace extracts was conducted using in vivo plant test system, such as Allium cepa, a sustainable method which can provide valuable information on the cytotoxic and genotoxic effect of extracts from natural sources in relation with their phytochemical composition. In this view, different aqueous and hydroethanolic extractions from black and red currant pomaces were carried out. For revealing the differences in the phytochemical profile of the studied extracts, rapid, efficient and easy-to-operate analytical techniques such as colorimetry, UV-Vis spectrometry and electrometry were used. Cytogenetic analysis of pomace extracts was achieved using Allium cepa test by scoring the mitotic index, the limit value of cytotoxicity, the phase index of mitosis and presence of chromosomal aberrations for all samples and comparing to the control (tap water). The results have shown that the cytogenetic response depends both on the type of pomaces and on the experimental extraction conditions. Compared to the control sample (22.58 %), a decrease in mitotic index for each analyzed sample was observed. As well, an increase in the cells with chromosomal aberrations was detected in onion root tips exposed to the tested extracts compared to the control (0.69 %). The lowest value of mitotic index (12.44 %) and the highest value of chromosomal aberrations (1.91 %) were recorded from the root tip cells of onion bulbs exposed to the hydroalcoholic extract obtained from red currants pomace using water/ethanol (60:40 v/v). Even if the mitotic index decreased, the limit value of cytotoxicity was higher than 50 %, level considered as a sublethal condition for the organisms. At the same time, the increase of the frequency of chromosomal aberrations is not so significant, and in correlation with the cytotoxicity limit, it does not indicate a genotoxic effect on onion cells. Following these results, it can be concluded that black and red currant pomace extracts can be safely used for possible therapeutic benefits.

Evaluation with physiological, cytogenetic, biochemical and anatomical aspects of dose-dependent inhibitory activity of jasmonic acid, a lipidderived growth regulator, in onion root tip cells

The effects of different doses of exogenous jasmonic acid (JA), were investigated on physiological parameters such as germination percentage, root length, root number and fresh weight, cytogenetic parameters such as mitotic index (MI), micronucleus (MN), frequency and chromosome aberration (CA), biochemical parameters as such superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA) and proline (PR) contents of Allium cepa L., known as onion. In addition, the changes in the root anatomical structures of the bulbs were examined under the microscope by taking cross-sections. Onion bulbs were divided into four groups as one control (C) and three treatments. The bulbs of the C group were kept in cuvettes containing tap water for 7 days and the bulbs of the treatment groups were kept in cuvettes containing 100 µM, 250 µM and 500 µM JA. JA administration caused a decrease in all physiological parameters examined, an increase in the frequency of MN and CA, and a reduce in MI compared to group C. In addition, the mentioned application caused a dosedependent increase in CAT and SOD activities and MDA and PR contents compared to group C. Moreover, 500 µM JA, the highest application dose, caused quite significant damages such as giant cell nucleus, unclear vascular tissue, binuclealar cell and accumulation of some substances in the cells of the epidermis/cortex in root anatomical structure of the bulbs. In summary, it was concluded that JA is a chemical with inhibitory functions and the Allium cepa test is a useful bioindicator for monitoring these effects.

Physiological and toxicological response of Microcystis aeruginosa BCCUSP232 exposed to Salvinia auriculata extracts

ABSTRACT Microcystis aeruginosa is one of the most predominant freshwater bloom-forming cyanobacterium found globally which is capable of producing toxic secondary metabolites including microcystins that might intoxicate animals and humans when contaminated water or food is ingested. Salvinia auriculata Aubl is one of the plants that might possess bioactive compounds capable of controlling growth and reproduction of M. aeruginosa. The present study aimed to determine the presence of bioactive compounds in S. auriculata extracts and determine alterations occurred in growth and reproduction of M. aeruginosa when exposed to these plant extracts. In addition, this investigation aimed to examine the influence of S. auriculata on antioxidant enzymes detected in M. aeruginosa. The results obtained demonstrated that the aqueous and ethanolic extracts of S. auriculata presented potential for control of cyanobacteria populations, exhibiting algicidal action on M. aeruginosa as well as interfering in antioxidant enzymes activities and parameters associated with oxidative stress. Phytochemical analyses demonstrated the presence of polyphenols and flavonoids content in both extracts. In addition, application of S. auriculata extracts did not produce cytogenotoxicity and/or mutagenicity utilizing Allium cepa test. Therefore, further studies are needed in order to identify and characterize the compounds responsible for these effects on M. aeruginosa and provide information regarding the possible application of S. auriculata in the treatment of drinking water.

Genotoxicity detected during cyanobacteria bloom in a water supply reservoir

The aquatic ecosystems are susceptible to cyanobacterial blooms due to the eutrophication of water bodies caused by human activities. In this study, phytoplankton and cyanotoxins analysis, as well as cellular and genetic biomarkers of toxicity (Allium cepa test - higher plant test system), were evaluated in water samples of Alagados Reservoir during a cyanobacterial bloom in South Brazil. The water samples were collected during the wet season at two sites in the Reservoir. Paralytic shellfish toxins (PSTs) were detected in both samples (sites 1 and 2); however, the levels of PSTs were higher in site 1. Gonyautoxin 2 was the major cyanotoxin found in the Reservoir. Both samples were able to induce cytotoxic effects (reduced Mitotic Index) and damage the genetic material (i.e., increased frequencies of chromosome aberration and micronuclei) of meristematic cells of A. cepa. The cellular and genetic damages were higher in the sample site 1, wherein high levels of PSTs were verified. Thus, our findings suggested that cyanotoxins-contaminated waters may damage the genetic material of living organisms, and therefore this group of contaminants should be assessed for their potential genotoxicity.

Toxicity and Phytochemical Analysis of Five Medicinal Plants

Recent studies have shown that long-term uses of herbs have been associated with a rise in morbidity and mortality rates. While most researches are focused on bioactivity investigations, the toxicity of many plants has not been reported. There is a paucity of data on the potential toxicity of the following plants: Harungana madagascariensis (HM), Pterocarpus osun (PO), Phoenix dactylifera (PD), Annona muricata (AM), and Rutidea parviflora (RP). To evaluate the toxicity of the above-mentioned plants; two tests were employed namely: The Brine shrimp lethality test (BSLT) and the Allium cepa test. A correlation between the oral acute toxicity assay in mice and the LC50 obtained from BSLT has been established. Allium cepa test measures the potential genotoxic effects of plant extracts exerted on the root meristem of A. cepa (onions). Plant extracts were administered in concentrations ranging from 100 to 2500 µg/ml to the A. cepa for 72 h to obtain their Mitotic Indices (MI) and EC50. Results of the MI at 2500 µg/ml for HM, PO, PD, AM, and RP were 3.75, 4.96, 5.96, 6.10, and 6.71 while 281.81, 398.11, 501.19, 630.96, and 707.9 µg/ml were obtained as the respective EC50 values. Furthermore, 10-1000 mcg/ml concentrations were administered in the BSLT and the obtained LC50 values were 116.3, 250, 581.5, 581.5, and 750 µg/ml. The toxicity result demonstrated that the five plants were moderately toxic, with RP exhibiting minimal toxicity values and thus potentially having a good safety profile. The phytochemical screening of these plants revealed the presence of some pharmacologically important classes of compounds that are abundant. Several bioactive and toxic compounds were identified in the GC-MS analysis for some of the plants.

Quantitative assessment of paclitaxel in Taxus wallichiana zucc. bark using HPTLC densitometry: Insights for germplasm selection and toxicological studies

Taxus wallichiana Zucc. [Synonym: Taxus baccata subsp. Wallichiana (Zucc.) Pilg.] or Himalayan Yew belonging to the evergreen gymnosperm family Taxaceae, grows in temperate regions and has been used in traditional medicines to treat cancer, bronchial disorders, colds, cough, etc. The plant is known to contain paclitaxel (PTX), a key bioactive component with anticancer properties. High-performance thin layer chromatography (HPTLC) was used in the present study to estimate paclitaxel from the bark of T. wallichiana obtained from 15 different geographical locations in the eastern and western Himalayas of India. HPTLC method was determined to be a reliable and efficient technique for the quantification of paclitaxel, offering rapid outcomes.The bark materials of T. wallichiana chemotype obtained from Sikkim (Samanden) contained the highest concentration of paclitaxel, followed by West Bengal (Sandakphu), Arunachal Pradesh (Dibang Valley), and respectively. The present study also examined the cytotoxicity and genotoxicity of the aqueous bark extract of the TW1 chemotype at various doses on the meristematic cells derived from the roots of Allium cepa L.. The bark extracts demonstrate considerable cytotoxic and mitotic properties. This study could be useful to the pharmaceutical industry for selecting elite (high paclitaxel-producing) chemotypes of T. wallichiana. The toxicological evaluation of the T. wallichiana chemotypes (using Allium cepa test) could also help to determine the genotoxic properties of the bark extract.

Antigenotoxic potential of the aqueous basidiocarp extracts of Calocybe indica

Calocybe indica, commonly known as the milky white mushroom, is an edible mushroom native to India. The antigenotoxic potential of the aqueous basidiocarp extract (25 g/L and 50 g/L) of C. indica was investigated using the Allium cepa test system. Meristematic cells of root tips treated with 100% Coca-Cola served as a positive control. A significant reduction in the mitotic index (7.83 ± 0.60) was observed in the positive control (100% Coca-Cola) compared to the negative control. The highest percentage (65.25 ± 4.58) of chromosomal abnormalities was observed in the positive control. Chromosomal abnormalities were significantly reduced in root tip cells treated with Coca-Cola followed by treatment with basidiocarp extracts such as 25 g/L and 50 g/L (30.32 ± 4.44 and 14.20 ± 2.41, respectively). Coca-Cola induced chromosomal abnormalities were reduced by treatment with basidiocarp extracts, demonstrating the antimutagenic potential of C. indica. The present study indicates that the aqueous basidiocarp extracts of C. indica have anti-genotoxic effects. The clastogenic abnormalities caused by Coca-Cola are competently restored in the root meristem cells of A. cepa treated with the extract.

Evaluation of Genotoxic and Cytotoxic Activities of Leaf and Seed Extracts of Telfairia occidentalis

Telfairia occidentalis Hook (cucurbitaceae) leaves and seeds, which are used as vegetable in the preparation of soups and as medicine traditionally to treat various diseases by the Ibibios were investigated for genotoxic and cytotoxic effects using Allium cepa test. The effects of the leaves and seeds extracts on the root meristem cells of Allium cepa were investigated using onion bulbs exposed to 2.5 mg/mL, 5 mg/mL, and 10 mg/mL concentrations of the extracts for macroscopic and microscopic analysis. Tap water was used as a negative control and Methotrexate (0.1 mg/ml) was used as a positive control. There was statistically significant (p&lt; 0.05) inhibition of root growth depending on concentration by the extracts when compared with the negative control group. All the tested extracts were observed to have cytotoxic effects on cell division in A. cepa. The extracts-induced chromosomal aberrations and micronuclei (MNC) formations in A. cepa root tip cells were significant (p&lt;0.05) when compared with control group. The extracts treatment further induced cell death, ghost cells, cells membrane damage, and binucleated cells. The seed extract was found to have a higher cytotoxic and genotoxic effects than the leaf extract. These results suggest that the leaf and seeds extract of Telfairia occidentalis possess cytotoxic and genotoxic effects on A. cepa.

Cytological assessments and a methodological approach to observe the apoptotic effect of nickel sulfate (NiSO4) on Allium cepa L. root germination using EB/AO fluorescence staining

Ethidium bromide–acridine orange (EB/AO) is one of the fastest, most economically valid methods for separating living and dead cells in plant root tips. The present study aimed to investigate the apoptotic effect, nuclear abnormalities and cell division index using an Allium cepa test assay with EB/AO staining and an ImageJ program. Nickel sulphate (NiSO4) concentrations (1.75, 3.5, 7 and 14 ppm) were exposed to the root germination of A. cepa for 48 and 72 h to observe mitotic abnormalities, cytotoxic effects and 5-day exposure to apoptotic effects. It was found that amorphous nuclei, vacuolisation and C-mitosis were the most observed abnormalities, which increased at 14 ppm NiSO4 exposure after 48 and 72 h. The total number of nuclear abnormalities significantly increased at all concentrations and exposure periods. The ImageJ program was used to determine apoptosis rates. The data obtained showed that high concentrations of NiSO4 caused significantly more cell death in root tips than in the control group, resulting in root growth inhibition and increased apoptosis with increased concentration and exposure period. NiSO4 caused toxic activity on root growth, identified as apoptosis, especially at the cortex and vascular region in the root tips. The study found affected damaged and apoptotic areas on the root tips using an EB/AO staining method, which can be used as a marker of damaged tissue areas.

The cytotoxic, genotoxic and mitotoxic effects of Atractylis gummifera extract in vitro.

The Mediterranean thistle Atractylis gummifera L. (Asteraceae; AG) has diterpenoid glucosides; atractyloside and carboxyatractyloside that interact with mitochondrial protein adenine nucleotide translocator (ANT) and resulted in ATP inhibition. Despite its well-known toxicity, acute poisonings still occur with this plant. Although most symptoms are attributed to ANT and diterpenoids interaction, in-depth investigation of the effects of AG extract on various cellular processes has not been performed. We tested in vitro induction of mitochondrial permeability transition pore (MPTP) opening in bovine liver mitochondria and evaluated its cytotoxicity and genotoxicity using Allium cepa test. Cell division, mitotic index (MI) and total chromosomal and mitotic aberrations (TAs), that all seem potentially affected by ATP shortage, were studied in root cells of Allium cepa exposed to Atractylis gummifera extract. With the two different doses of two purified AG fractions, stronger induction of MPTP was observed compared to the induction with the standard pure atracyloside. Aqueous AG extract exerted inhibition root growth in A. cepa at 6 different doses. The TAs was increased in a dose-dependent manner too, while mitotic index was decreased at the same doses. Evaluation of mitotic phases revealed mitodepressive effect of AG on A. cepa roots. this work highlights cellular and mitochondrial adverse effects of Atractylis gummifera extracts. A purified fraction that likely corresponds to ATR derivatives induces MPTP opening leading to swelling of mitochondria and its dysfunction. Allium cepa test provides the evidence for A. gummifera genotoxicity and cytotoxicity.