In Vitro Effects of Streptomyces tyrosinase on the Egg and Adult Worm of Toxocara vitulorum
Background: Several species of streptomycetes, saprophytic bacteria found widely distributed in soil, water and plants, produce bioactive compounds such as intra and extracellular hydrolases including lytic enzymes which reflecting on their importance in the biological control of insects and parasites. This study assessed the in vitro effects of Streptomyces tyrosinase, produced from Streptomyces spp. isolated from Egyptian soil, on animal-parasitic nematode Toxocara vitulorum, in terms of egg development and adult worm’s cuticular structure, and as an alternative strategy to alleviate this infection.
Methods: This study was conducted at the National Research Centre, Egypt in 2018. Five different concentrations of tyrosinase, ranged from 1%-30% were tested against the development of T. vitulorum eggs. The concentration induced the highest inhibitory activity was tested against adult T. vitulorum cuticle, which is essential for the protective and nutritive functions. The results were compared with those observed in the egg development and worm cuticle following incubation in Streptomyces protease (as a reference enzyme).
Results: Compared to Streptomyces protease, higher inhibitory activity on T. vitulorum egg development and extreme cuticular alterations of the treated adult worms had been observed following 24 h exposure to Streptomyces tyrosinase. Once the cuticle had been damaged, the enzyme would be able to penetrate deeper into the internal tissues of the nematode and caused more widespread disruption.
Conclusion: The current study could offer a promising bio-control agent, Streptomyces tyrosinase, against T. vitulorum alternative to the more expensive synthetic anthelmintics.
2. Hussain AA, Mostafa SA, Ghazal SA, Ibrahim SY. Studies on antifungal antibiotic and bioinsecticidal activities of some actinomycete isolates. Afr J My-col Biotechnol. 2002; 10: 63-80.
3. Tian B, Yang J, Zhang KQ. Bacteria used in the biological control of plant-parasitic nematodes: populations, mechanisms of action, and future pro-spects. FEMS Microbiol Ecol. 2007; 61: 197-213.
4. Tunga R, Shrivastava B, Banerjee R. Purification and characterization of a protease from solid state cultures of Aspergillus parasiticus. Process Biochem. 2003; 38: 1553-1558.
5. Harrison RL, Bonning BC. Proteases as insecticidal agents. Toxins (Basel) 2010; 2: 935-953.
6. El-Gammal EW, Shalaby HA, Ashry HM, El-Diwany AI. In vitro action of Streptomyces griseolus pro-teases as bio-control on Fasciola gigantica eggs. J Bac-teriol Parasitol. 2014; 5:4
7. Fairhead M, Thöny-Meyer L. Cross-linking and immobilisation of different proteins with recombi-nant Verrucomicrobium spinosum tyrosinase. J Biotech-nol. 2010; 150: 546-551.
8. Martorell MM, Pajot HF, Rovati JI, Figueroa LIC. Optimization of culture medium composition for manganese peroxidase and tyrosinase production during Reactive Black 5 decolourization by the yeast Trichosporon akiyoshidainum. Yeast. 2012; 29: 137-144.
9. Saad MM, Mostafa EE, Hassbo AA, Selim MH. Production, partial purification of tyrosinase pro-duced by Streptomyces ghanesis SAH1_CWMSG and application as anticancer. Bull NRC. 2017; 41: 165-176.
10. Perry BD, Randolph TF. Improving assessment of the economic impact of parasitic diseases and their control in production animals. Vet Parasitol. 1999; 84: 145-168.
11. Yadav CL, Kumar RR, Vatsya S, Garg R, Baneerjee PS. Epidemiological studies on gastrointestinal nematodosis in cattle and buffaloes. J Vet Parasitol. 2008; 22: 57-62.
12. Roberts JA. The extra-parasitic life cycle of Toxocara vitulorum in the village environment of Sri Lanka. Vet Res. 1989; 13: 377-388.
13. Borecka A, Gawor J, Niedworok M, Sordyl B. [Occurrence of Toxocara spp. eggs in household environment of children with diagnosed toxocaria-sis in Łódź voivodeship]. Wiad Parazytol. 2010; 56: 141-144. [Article in Polish]
14. Roldán WH, Espinoza YA, Huapaya PE, Jiménez S. Diagnosis of human toxocariasis. Rev Peru Med Exp Salud Publica. 2010; 27: 613-620.
15. Awad HM, Mostafa EE, Saad MM, Selim MH, Hassan HM. Partial purification and characterization of extracellular protease from a halophilic Streptomyces pseudogrisiolus NRC-15. Indian J Biochem Biophys. 2013; 50: 305-311.
16. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol rea-gent. J Biol Chem. 1951; 193: 265-275.
17. Oshima T. Standardisation of techniques for infect-ing mice with Toxocara canis and observations on the normal migration routes of the larvae. J Parasitol. 1961; 47: 652-656.
18. Hanser E, Mehlhorn H, Hoeben D, Vlaminck K. In vitro studies on the effects of flubendazole against Toxocara canis and Ascaris suum. Parasitol Res. 2002; 89: 63-74.
19. Bancroft JD, Stevens A, Turner DR. Theory and Practice of Histological Techniques, 4th ed. Churchill Livingstone, New York/London/San Francisco/Tokyo, 1996.
20. Yang J, Liang L, Li J, Zhang K. Nematicidal en-zymes from microorganisms and their applications. Appl Microbiol Biotechnol. 2013; 97: 7081-7095.
21. Wharton DA. Nematode egg-shells. Parasitol. 1980; 81: 447-463.
22. Cox GN, Kusch M, Edgar RS. Cuticle of Caeno-rhabditis elegans: its isolation and partial characteriza-tion. J Cell Biol. 1981; 90: 7-17.
23. Lopez-Llorca LV. Purification and properties of extracellular proteases produced by the nematoph-agous fungus Verticillium suchlasporium. Can J Micro-biol. 1990; 36: 530-537.
24. Tunlid A, Rosen S, Ek B, Rask L. Purification and characterization of an extracellular serine protease from the nematode-trapping fungus Arthrobotrys oli-gospora. Microbiol. 1994; 140: 1687-1695.
25. Qiuhong N, Xiaowei H, Baoyu T, Jinkui Y, Jiang L, Lin Z, Keqin Z. Bacillus sp. B16 kills nematodes with a serine protease identified as a pathogenic fac-tor. Appl Microbiol Biotechnol. 2006; 69: 722-730.
26. Yang J, Huang X, Tian B, Wang M, Niu Q, Zhang K. Isolation and characterization of a serine prote-ase from the nematophagous fungus, Lecanicillium psalliotae, displaying nematicidal activity. Biotechnol Lett. 2005; 27: 1123-1128.
27. Huang XW, Zhao NH, Zhang KQ. Extracellular enzymes serving as virulence factors in nematoph-agous fungi involved in infection of the host. Res Microbiol. 2004; 155: 811-816.
28. Yang JK, Tian BY, Liang LM, Zhang KQ. Extra-cellular enzymes and the pathogenesis of nema-tophagous fungi. Appl Microbiol Biotechnol. 2007; 75: 21-31.
29. Anwar S, Ali B, Qamar F, Sajid I. Insecticidal activi-ty of actinomycetes isolated from salt range, Paki-stan against mosquitoes and red flour beetle. Pak J Zool. 2014; 46: 83-92.
30. Braga FR, Araújo JV, Campos AK, et al. Ob-servação in vitro da ação dos isolados fúngicos Duddingtonia flagrans, Monacrosporium thaumasium e Ver-ticillium chlamydosporium sobre ovos de Ascaris lumbri-coides (Lineu, 1758). Revista da Sociedade Brasileira de Medicina Tropical 2007; 40: 356-358.
31. Araújo JV, Santos MA, Ferraz S. Efeito ovicida de fungos nematófagos sobre ovos embrionados de Toxocara canis. Arquivo Brasileiro de Medicina Veter-inária e Zootecnia 1995; 47: 37-42.
32. Bird AF. Chemical composition of the nematode cuticle. Observations of the whole cuticle. Exp Par-asitol. 1956; V: 350-358.
33. Popa C, Bahrim G. Streptomyces tyrosinase: pro-duction and practical applications. Innov Rom Food Biotechnol. 2011; 8: 1-7.
34. Claus H, Filip Z. Behavior of phenoloxidases in the presence of clays and other soil related adsorbents. Appl Microbiol Biotechnol. 1988; 28: 506-511.
35. Claus H, Filip Z. Enzymatic oxidation of some substituted phenols and aromatic amines, and the behavior of some phenoloxidases in the presence of soil related adsorbents. Water Sci Technol. 1990; 22: 69-77.
36. Jordan AM, Khan TH, Malkin H, Osborn HM, Photiou A, Riley PA. Melanocyte directed enzyme prodrug therapy (MDEPT). Development of sec-ond generation prodrugs for targeted treatment of malignant melanoma. Bioorg Med Chem. 2001; 9: 1549-1558.
37. Shalaby HA, El Namaky AH, Khalil FA, Kandil OM. Efficacy of methanolic extract of Balanites ae-gyptiaca fruits on Toxocara vitulorum. Vet Parasitol. 2012; 183: 386-392.
38. Halton DW. Microscopy and the helminth parasite. Micron. 2004; 35: 361-390.
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