Anti-Blastocystis Activity In Vitro of Egyptian Herbal Extracts (Family: Asteraceae) with Emphasis on Artemisia judaica
Abstract
:1. Introduction
2. Materials and Methods
2.1. Source of Faecal Samples
2.2. Cultivation of Blastocystis spp. In Vitro
2.3. Molecular Identification of Blastocystis spp. Subtypes
2.4. Collection of Plant Materials
2.5. Plants Extraction
2.6. Use of Metronidazole as a Reference Anti-Protozoal Drug
2.7. In Vitro Exposure of Blastocystis spp. to Egyptian Herbal Extracts
2.8. Counting the Treated Blastocystis in Suspensions to Examine the Efficacy of the Herbal Extracts Tested
2.9. Fractionation of A. judaica Extract
2.10. Statistics
3. Results
3.1. Blastocystis spp. Isolates and Its Molecular Identification
3.2. Anti-Blastocystis Activity
3.3. Subtype-Dependent Variations of Blastocystis
3.4. Identification of Active Fractions in A. judaica
4. Discussion
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
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Isolate No. | Age of Patient | Sex | Symptoms | Since How Long | Stool Consistency | Microscopic Examination | Molecular Subtyping |
---|---|---|---|---|---|---|---|
Isolate 1 | 6 years | Male | Diarrhoea | 3.5 months | Semi-formed | 1–2 Blastocystis forms/field | ST1 |
Isolate 2 | 8 years | Female | Abdominal pain | 7 months | Formed | 4–5 Blastocystis forms/field | ST3 |
Subtypes | STs Primer Sets | Primers | Expected PCR Size | PCR Results of This Study’s Isolates |
---|---|---|---|---|
ST1 | SB83 | F 5′-GAAGGACTCTCTGACGATGA-3′ R 5′-GTCCAAATGAAAGGCAGC-3′ | 351 bp | ST1 |
ST2 | SB340 | F 5′-TGTTCTTGTGTCTTCTCAGCTC-3′ R 5′-TTCTTTCACACTCCCGTCAT-3′ | 704 bp | - |
ST3 | SB227 | F 5′-TAGGATTTGGTGTTTGGAGA-3′ R 5′-TTAGAAGTGAAGGAGATGGAAG-3′ | 526 bp | ST3 |
ST4 | SB337 | F 5′-GTCTTTCCCTGTCTATTCTGCA-3′ R 5′-AATTCGGTCTGCTTCTTCTG-3′ | 487 bp | - |
ST5 | SB336 | F 5′-GTGGGTAGAGGAAGGAAAACA-3′ R 5′-AGAACAAGTCGATGAAGTGAGAT-3′ | 317 bp | - |
ST6 | SB332 | F 5′-GCATCCAGACTACTATCAACATT-3′ R 5′-CCATTTTCAGACAACCACTTA-3′ | 338 bp | - |
ST7 | SB155 | F 5′-ATCAGCCTACAATCTCCTC-3′ R 5′-ATCGCCACTTCTCCAAT-3′ | 650 bp | - |
Plant Species | MIC90 of Blastocystis in Culture † | Degree of Blastocystis elimination * | ||||||
---|---|---|---|---|---|---|---|---|
Conc. (µg/mL) | Blastocystis ×104 cells/mL after 24 h (% Inhibition) | Blastocystis ×104 cells/mL after 48 h (% Inhibition) | Blastocystis ×104 cells/mL after 72 h (% Inhibition) | |||||
A. fragrantissima | 250 | 51.1 | (41.7) | 15 | (80.3) | 44.5 | (48.6) | ++ |
500 | 46.8 | (46.7) | 13.9 | (81.8) | 39.3 | (54.7) | ++ | |
1000 | 22.9 | (73.9) | 13.5 | (82.3) | 17.75 | (79.4) | + | |
2000 | 5.9 | (93.3) | 3.8 | (95) | 3.75 | (95.6) | - | |
4000 | 0 | (100) | 0 | (100) | 0 | (100) | - | |
E. spinosus | 250 | 30.9 | (64.8) | 19.3 | (74.7) | 45.6 | (47.3) | ++ |
500 | 33.8 | (61.5) | 18.5 | (75.8) | 41.9 | (54.7) | ++ | |
1000 | 23.5 | (73.2) | 8.9 | (88.2) | 41.5 | (52) | ++ | |
2000 | 12 | (86.3) | 29.3 | (61.6) | 44.5 | (48.6) | ++ | |
4000 | 0 | (100) | 0 | (100) | 0 | (100) | - | |
A. judaica | 250 | 41 | (53.3) | 11.1 | (85.5) | 36 | (58.4) | ++ |
500 | 23.4 | (73.4) | 7.25 | (90.4) | 13 | (85) | + | |
1000 | 23.9 | (72.8) | 6 | (92.1) | 13.9 | (83.9) | + | |
2000 | 2.9 | (96.7) | 0.4 | (99.4) | 0.5 | (99.3) | - | |
4000 | 0 | (100) | 0 | (100) | 0 | (100) | - | |
MTZ (Reference drug) | 5 | 14 | (84) | 12.5 | (83.6) | 9.5 | (89) | + |
10 | 2.5 | (97.2) | 7.6 | (90) | 14.75 | (82.8) | + | |
20 | 0.5 | (99.4) | 0 | (100) | 0 | (100) | - |
Fractions of A. judaica | Subtypes of Blastocystis | |||
---|---|---|---|---|
ST1 | ST3 | |||
EtOAc | 100 | a | 100 | a |
DCM | 100 | a | 100 | a |
n-BuOH | 92.4 | c | 95 | b |
n-hexane | 100 | a | 100 | a |
Water-soluble fraction * | 95 | b | 94 | b |
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Mokhtar, A.B.; Ahmed, S.A.; Eltamany, E.E.; Karanis, P. Anti-Blastocystis Activity In Vitro of Egyptian Herbal Extracts (Family: Asteraceae) with Emphasis on Artemisia judaica. Int. J. Environ. Res. Public Health 2019, 16, 1555. https://doi.org/10.3390/ijerph16091555
Mokhtar AB, Ahmed SA, Eltamany EE, Karanis P. Anti-Blastocystis Activity In Vitro of Egyptian Herbal Extracts (Family: Asteraceae) with Emphasis on Artemisia judaica. International Journal of Environmental Research and Public Health. 2019; 16(9):1555. https://doi.org/10.3390/ijerph16091555
Chicago/Turabian StyleMokhtar, Amira B., Shahira A. Ahmed, Enas E. Eltamany, and Panagiotis Karanis. 2019. "Anti-Blastocystis Activity In Vitro of Egyptian Herbal Extracts (Family: Asteraceae) with Emphasis on Artemisia judaica" International Journal of Environmental Research and Public Health 16, no. 9: 1555. https://doi.org/10.3390/ijerph16091555