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Antiparasitic potentiality of ethanol and methanol extracts of Azadirachta indica leaf for eggs and copepodid stage of Argulus japonicus: in vitro study.

ABSTRACT: In the present study, eggs and copepodid stages of Argulus japonicus were treated with ethanol and methanol extract of Azadirachta indica (neem) leaf and its antiparasitic efficacy (AE %) was determined. The experiments were performed in triplicate along with the positive (2% DMSO) and negative (without DMSO and extract) control groups. The reduced cumulative hatching percentage of eggs by 13% (in ethanolic) and 17% (in methanolic) extract of neem leaf at 1.5 g L^-1 was obtained during 15-day exposure compared to the control group showing 70-85% eggs hatching. The AE of 100% for ethanolic and 91.66% for methanolic extract against the copepodid stage was found at 1.25 and 1.5 g L^-1 respectively in 6 h. The histological analysis of the eggs showed the undifferentiated decaying mass of cells with extensively damaged eggs when treated with ethanolic extract of neem leaf. Further, severe degeneration in the branchial region, digestive tract and eye cells was observed in the copepodids treated with ethanol extract than the methanol extract. The terpenoids a potential antiparasitic compound of ethanolic extract produced more AE than the methanolic extract. Thus, the ethanolic extract of neem leaf can be potentially utilized as a natural parasiticide to disrupt the egg and other life phases of A. japonicus.

SUBMITTER: Kumari P

PROVIDER: S-EPMC8368549 | biostudies-literature | 2021 Sep

REPOSITORIES: biostudies-literature

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Antiparasitic potentiality of ethanol and methanol extracts of Azadirachta indica leaf for eggs and copepodid stage of Argulus japonicus: in vitro study.

Kumari Pushpa P Kumar Saurav S Deo Ashutosh D AD Rajendran K V KV Raman R P RP

Journal of parasitic diseases : official organ of the Indian Society for Parasitology 20210217 3

In the present study, eggs and copepodid stages of Argulus japonicus were treated with ethanol and methanol extract of Azadirachta indica (neem) leaf and its antiparasitic efficacy (AE %) was determined. The experiments were performed in triplicate along with the positive (2% DMSO) and negative (without DMSO and extract) control groups. The reduced cumulative hatching percentage of eggs by 13% (in ethanolic) and 17% (in methanolic) extract of neem leaf at 1.5 g L-1 was o ...[more]

PMID: 34475659

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Project description:BackgroundAntibiotic resistances of pathogens and breast cancer warrant the search for new alternative strategies. Phytoextracts can eradicate microbe-borne diseases as well as cancer with lower side effects compared to conventional antibiotics.AimUnripe and ripe Azadirachta indica (neem) seed extracts were explored as potential antibiofilm and anticancer agents in combating multidrug-resistant infectious bacteria as well as anticancer agents against the MDR breast cancer cell lines.MethodsShed-dried neem seeds (both unripe and ripe) were pulverized and extracted using methanol. The chemical components were identified with FTIR and gas chromatography - mass spectrometry. Antibiofilm activity of neem seed extracts were assessed in terms of minimum biofilm inhibitory concentration (MBIC), minimum biofilm eradication concentration (MBEC), and fluorescence microscopic studies on Staphylococcus aureus and Vibrio cholerae. Bacterial cells were studied by fluorescence microscopy using acridine orange/ethidium bromide as the staining agents. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values were evaluated to observe the antibacterial activities. Cytotoxicity of the extracts against human blood lymphocytes and the anticancer activity against drug-resistant breast cancer cell lines were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and fluorescence-activated cell sorting (FACS) studies.Results4-Ethyl-2-hydroxy-2-cyclopentene-1-one, phthalic acid, and 2-hexyl-tetrahydro thiophane were the major compounds in unripe neem seed, whereas 3,5-dihydroxy-6-methyl-2,3-dihydro-4-H-pyran-4-one and 4-ethylbenzamide were predominant in ripe neem seed. Triazine derivatives were also common for both the extracts. MBIC values of unripe and ripe neem seed extracts for S. aureus are 75 and 100 µg/mL, respectively, and for V. cholerae, they are 100 and 300 µg/mL, respectively. MBEC values of unripe and ripe seed extracts are 500 and 300 µg/mL, respectively for S. aureus and for V. cholerae the values are 700 and 500 µg/mL, respectively. Fluorescence microscopic studies at 16 and 24 h, after bacterial culture, demonstrate enhanced antibiofilm activity for the ripe seed extract than that of the unripe seeds for both the bacteria. MTT assay reveals lower cytotoxicity of both the extracts towards normal blood lymphocytes, and anticancer activity against breast cancer cell line (MDA-MB-231) with superior activity of ripe seed extract. FACS studies further supported higher anticancer activity for ripe seed extract.ConclusionsMethanolic extract of neem seeds could substantially inhibit and eradicate biofilm along with their potent antibacterial and anticancer activities. Both the extracts showed higher antibiofilm and antibacterial activity against S. aureus (gram-positive) than V. cholerae (gram-negative). Moreover, ripe seed extract showed higher antibiofilm and anticancer activity than unripe extracts.

Dataset Information

Antiparasitic potentiality of ethanol and methanol extracts of Azadirachta indica leaf for eggs and copepodid stage of Argulus japonicus: in vitro study.

Publications

Antiparasitic potentiality of ethanol and methanol extracts of <i>Azadirachta indica</i> leaf for eggs and copepodid stage of <i>Argulus japonicus</i>: in vitro study.

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