Invertebrate Neuroscience

, 19:7 | Cite as

Assessment of the molluscicidal impact of extracted chlorophyllin on some biochemical parameters in the nervous tissue and histological changes in Biomphalaria alexandrina and Lymnaea natalensis snails

  • Amina M. IbrahimEmail author
  • Fayez A. Bakry
Original Article


Biomphalaria alexandrina and Lymnaea natalensis snails are the intermediate hosts of schistosomiasis and fasciolosis. The aim of the present study is to evaluate the molluscicidal activity of chlorophyll extract as a photodynamic substance against these snails and how it affected its tissues and the biological system. Chlorophyllin was extracted from deep-frozen Moringa oleifera leaves, and then it was transformed into water-soluble chlorophyllin. The present results showed that it had a molluscicidal activity on B. alexandrina snails (LC50 17.6 mg/l; LC90 20.9 mg/l) and L. natalensis snails (LC50 4.3 mg/l; LC90 6.8 mg/l). Exposing B. alexandrina snails to the sublethal concentrations (LC0, LC10, and LC25) resulted in a significant reduction in their survival rates. Regarding its effect on biochemical parameters, chlorophyllin significantly reduced the acetylcholinesterase activity, protein content, and alkaline and acid phosphatase activity in B. alexandrina nervous tissue compared to the control group. Histopathological changes occurred in the digestive gland of treated B. alexandrina snails where cells lost their nuclei, vacuolated, degenerated, and ruptured, and the lumen increased. Photosynthesizing materials like chlorophyllin are new approaches to control schistosomiasis and fasciolosis in developing countries by affecting their intermediate host. These materials were cheap and environmentally safe to replace the synthetic molluscicides for snail control.


Biomphalaria alexandrina Lymnaea natalensis Chlorophyllin Survival rate Enzymes Histopathological changes 


Author’s contributions

AMI and FAB conceived and designed the study. AMI performed the experiments and analyzed the data. AMI wrote the first draft, and FAB revised and edited it. Both authors read and approved the final manuscript.


The authors would like to thank the financial support of the internal project “103 M,” Theodor Bilharz Research institute, Giza, Egypt.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Human and animal rights

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. This article does not contain any studies with human participants performed by any of the authors.

Ethics approval and consent to participate

Ethical approval had been granted approval by the Ethics Committee of Theodor Bilharz Research Institute (TBRI).


  1. Abd El-Ghany AM, Abd El-Ghany NM (2017) Molluscicidal activity of Bacillus thuringiensis strains against Biomphalaria alexandrina snails. Beni Suef Univ J Basic Appl Sci 6:391–393. CrossRefGoogle Scholar
  2. Abdel-Ghaffar F, Ahmed AK, Bakry F, Rabei I, Ibrahim AM (2016) The impact of three herbicides on biological and histological aspects of Biomphalaria alexandrina, intermediate host of Schistosoma mansoni. Malacologia 59(2):197–210CrossRefGoogle Scholar
  3. Abou-Donia M (1978) Increased acid phosphatase activity in hens following an oral dose of leptophos. Toxicol Lett 2:199–203CrossRefGoogle Scholar
  4. Aruna P, Sreeramulu Chetty C, Chandramohan Naidu R, Swami KS (1979) Acid phosphatase activity in the Indian apple snail, Pila globosa (Swainson), during aestivation and starvation stress. Proc Anim Sci 88:363–365. CrossRefGoogle Scholar
  5. Beesley NJ, Williams DJL, Paterson S, Hodgkinson J (2017) Fasciola hepatica demonstrates high levels of genetic diversity, a lack of population structure and high gene flow: possible implications for drug resistance. Int J Parasitol 47:11–20. CrossRefPubMedPubMedCentralGoogle Scholar
  6. Bergmeyer H (1985) Methods of enzymatic analysis, methods of enzymatic analysis. Metabolites 3: lipids, amino acids and related compounds, vol 8. VCH Verlagsgesellschaft, WeinheimGoogle Scholar
  7. Brien RDO (1976) Acetylcholinesterase and its inhibition. In: Wilkinson CF (ed) Insecticide biochemistry and physiology. Plenum Press, New York, pp 271–273Google Scholar
  8. Chaturvedi D, Vinay D, Singh K (2017) Assessment the effect of photodynamic chlorophyllin on biochemical changes in the cerebral ganglion of snail lymnaea acuminata. Int J Pharma Sci Res 8:68–75Google Scholar
  9. Chontananarth T, Tejangkura T, Wetchasart N, Chimburut C (2017) Morphological characteristics and phylogenetic trends of trematode cercariae in freshwater snails from Nakhon Nayok Province, Thailand. Korean J Parasitol 55:47–54. CrossRefPubMedPubMedCentralGoogle Scholar
  10. DeRosa M (2002) Photosensitized singlet oxygen and its applications. Coord Chem Rev 233–234:351–371. CrossRefGoogle Scholar
  11. Elhadad HA, El-Habet BA, Azab RM et al (2018) Effect of Chlorophyllin on Biomphalaria alexandrina snails and Schistosoma mansoni Larvae. Int J Curr Microbiol Appl Sci 7:3725–3736. CrossRefGoogle Scholar
  12. Ellman GL, Courtney KD, Andres V, Featherstone RM (1961) A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 7:88–95. CrossRefGoogle Scholar
  13. Elsareh F, Abdalla R, Abdalla E (2016) The effect of aqueous leaves extract of Solenostemma argel (Del Hayne) on egg masses and neonates of Biomphalaria pfeifferi snails. J Med Plants 4:271–274Google Scholar
  14. Erzinger GS, Wohllebe S, Vollrath F et al (2011) Optimizing conditions for the use of chlorophyll derivatives for photodynamic control of parasites in aquatic ecosystems. Parasitol Res 109:781–786. CrossRefPubMedGoogle Scholar
  15. Erzinger GS, Souza SC, Pinto LH et al (2015) Assessment of the impact of chlorophyll derivatives to control parasites in aquatic ecosystems. Ecotoxicology 24:949–958. CrossRefPubMedGoogle Scholar
  16. Finney DJ (1971) Probit analysis, 3rd edn. Cambridge University Press, CambridgeGoogle Scholar
  17. Ibrahim AM, Abdalla AM (2017) Impact of Moringa oleifera seed aqueous extract on some biological, biochemical, and histological aspects of Biomphalaria alexandrina snails. Environ Sci Pollut Res 24:28072–28078. CrossRefGoogle Scholar
  18. Ibrahim AM, Migazi MG, Dexian ES (1977) Histochemical localization of alkaline phosphatase activity in the alimentary tract of the snail Marisa cornuarietis (L). Zool Soc Egypt Bull 26:94–105Google Scholar
  19. Jaiswal P, Kumar P, Singh VK, Singh DK (2010) Enzyme inhibition by molluscicidal components of Myristica fragrans Houtt. in the nervous tissue of snail Lymnaea acuminata. Enz Res. CrossRefGoogle Scholar
  20. Kiros G, Erko B, Giday M, Mekonnen Y (2014) Laboratory assessment of molluscicidal and cercariacidal effects of Glinus lotoides fruits. BMC Res Notes 7:1CrossRefGoogle Scholar
  21. Kumar N, Singh VK (2016) Effect of chlorophyllin bait on acetylcholinesterase and cytochrome oxidase activities in the nervous tissue of Lymnaea acuminata with exposure of sunlight and red light. Eur J Biol Res 6:254–259. CrossRefGoogle Scholar
  22. Lee J-H, Quan J-H, Choi I-W, Park GM, Cha GH, Kim HJ, Yuk JM, Lee YH (2017) Fasciola hepatica: infection status of freshwater snails collected from Gangwon-do (Province), Korea. Korean J Parasitol 55:95. CrossRefPubMedPubMedCentralGoogle Scholar
  23. Lowry O, Schagger H, Cramer WA, Vonjagow G (1994) Protein measurement with the folin phenol reagent. Anal Biochem 217:220–230. CrossRefGoogle Scholar
  24. Mahmoud M, Richter P, Kandil O et al (2013) Molluscicidal activity of chlorophyll extraction against the freshwater snails. J Coast Life Med 1:85–88. CrossRefGoogle Scholar
  25. Matozzo V, Tomei A, Marin MG (2005) Acetylcholinesterase as a biomarker of exposure to neurotoxic compounds in the clam Tapes philippinarum from the Lagoon of Venice. Mar Pollut Bull 50:1686–1693. CrossRefPubMedGoogle Scholar
  26. Moema EBE, King PH, Baker C (2008) Cercariae developing in Lymnaea natalensis Krauss, 1848 collected in the vicinity of Pretoria, Gauteng Province, South Africa. Onderstepoort J Vet Res 75:215–223CrossRefPubMedGoogle Scholar
  27. Mohamed SH, Saad AA (1990) Histological studies on the hermaphrodite gland of Lymnaea caillaudi and Biomphalaria alexandrina upon infection with certain larval trematodes. Egypt J Histol 13:47–53Google Scholar
  28. Ragheb M, El-Tayeb TA, El-Emam MA et al (2018) Fecundity, sex hormones and release of cercariae of Schistosoma mansoni in Biomphalaria alexandrina (Ehrenberg, 1831) treated with copper and magnesium chlorophyllin. Folia Malacol 26:17–24. CrossRefGoogle Scholar
  29. Rahman AKMA, Islam SKS, Talukder MH et al (2017) Fascioliasis risk factors and space-time clusters in domestic ruminants in Bangladesh. Parasit Vectors 10:228. CrossRefPubMedPubMedCentralGoogle Scholar
  30. Rahmani M, Csallany AS (1991) Chlorophyll and β-carotene pigments in moroccan virgin olive oils measured by high-performance liquid chromatography. J Am Oil Chem Soc 68:672–674. CrossRefGoogle Scholar
  31. Richter PR, Strauch SM, Azizullah A, Häder D-P (2014) Chlorophyllin as a possible measure against vectors of human parasites and fish parasites. Front Environ Sci 2:18. CrossRefGoogle Scholar
  32. Singh DK, Agarwal RA (1991) Action sites of cypermethrin, a synthetic pyrethroid in the snail Lymnaea acuminata. Acta Hydrochim Hydrobiol 19:425–430. CrossRefGoogle Scholar
  33. Singh RN, Kumar P, Singh VK, Singh DK (2010) Available online through Toxic effects of Deltamethrin on the levels of biochemical changes in the snail Lymnaea acuminata. J Pharm Res 3:1739–1742Google Scholar
  34. Soliman MFM (2008) Epidemiological review of human and animal fascioliasis in Egypt. J Infect Dev Ctries 2:182–189. CrossRefPubMedGoogle Scholar
  35. WHO (1965) Molluscicide screening and evaluation. Bull World Health Organ 33(4):567–581Google Scholar
  36. WHO (1983) Report of the Scientific working Group on Plant Molluscicide and guidelines for evaluation of plant molluscicides. Geneva: WHO (TDR/SCHSWE (4)/833) Google Scholar
  37. WHO (2014) Schistosomiasis. Fact Sheet No. 115Google Scholar
  38. WHO (2017) Schistosomiasis. Fact Sheet 115, updated Oct., 2017Google Scholar
  39. Wohllebe S, Richter R, Richter P, Häder D-P (2009) Photodynamic control of human pathogenic parasites in aquatic ecosystems using chlorophyllin and pheophorbid as photodynamic substances. Parasitol Res 104:593–600. CrossRefPubMedGoogle Scholar
  40. Wohllebe S, Richter P, Häder D-P (2012) Chlorophyllin for the control of Ichthyophthirius multifiliis (Fouquet). Parasitol Res 111:729–733. CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Environmental Research and Medical Malacology DepartmentTheodor Bilharz Research InstituteImbaba, GizaEgypt

Personalised recommendations