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Parasitology Research

, Volume 118, Issue 7, pp 2053–2063 | Cite as

Inhibitory activity of chitosan nanoparticles against Cryptosporidium parvum oocysts

  • Shahira A. AhmedEmail author
  • Heba S. El-Mahallawy
  • Panagiotis Karanis
Protozoology - Original Paper
  • 89 Downloads

Abstract

Cryptosporidium is a ubiquitous harsh protozoan parasite that resists many disinfectants. It remains viable and infective for a long time in water and food causing global outbreaks. Chitosan (the deacetylated chitin molecule) was used in its nanosuspension form to evaluate its effect against Cryptosporidium parvum. The experiments were performed in vitro in serial concentrations and confirmed in mice in vivo infectivity assay. Chitosan nanoparticles (Cs NPs) were toxic to Cryptosporidium oocysts. The effect appeared to decrease the number of Cryptosporidium oocysts and altered their content. The destruction rate of oocysts was dependent on the dose of chitosan and the time of exposure (P < 0.05). Higher doses of Cs NPs over a prolonged period exhibited a significantly higher destruction rate. Using staining and light microscopy, remarkable destructive changes were observed in the oocysts’ morphology. The minimal lethal dose for > 90% of oocysts was 3000 μg/ml, no mice infections in vivo were observed. The results in this study elucidate Cs NPs as an effective anti-cryptosporidial agent.

Keywords

Cryptosporidium Chitosan In vitro Nanoparticles Bioassay Activity 

Notes

Compliance with ethical standards

The protocols for sample collection, the laboratory animal housing and inoculations were reviewed and approved by the Scientific Research Committee and Bioethics Board of Suez Canal University, Faculty of Medicine, Ismailia, Egypt.

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Abebe LS, Su Y-H, Guerrant RL, Swami NS, Smith AS (2015) Point-of-use removal of Cryptosporidium parvum from water: Independent effects of disinfection by silver nanoparticles and silver ions and by physical filtration in ceramic porous media. Environ Sci Technol 49:12958–12967CrossRefGoogle Scholar
  2. Ahmed SA, Guerrero Flórez M, Karanis P (2018) The impact of water crises and climate changes on the transmission of protozoan parasites in Africa. Pathog Glob Health 112:281–293CrossRefGoogle Scholar
  3. Ahmed SA, Karanis P (2018a) An overview of methods/techniques for the detection of Cryptosporidium in food samples. Parasitol Res 117:629–653CrossRefGoogle Scholar
  4. Ahmed SA, Karanis P (2018b) Comparison of current methods used to detect Cryptosporidium oocysts in stools. Int J Hyg Environ Health 221:743–763CrossRefGoogle Scholar
  5. Armson A, Meloni BP, Reynoldson JA, Thompson RCA (1999) Assessment of drugs against Cryptosporidium parvum using a simple in vitro screening method. FEMS Microbiol Lett 178:227–233CrossRefGoogle Scholar
  6. Baldursson S, Karanis P (2011) Waterborne transmission of protozoan parasites: review of worldwide outbreaks - an update 2004-2010. Water Res 45:6603–6614CrossRefGoogle Scholar
  7. Bell IR, Ives JA, Jonas WB (2014) Non-linear effects of nanoparticles: biological variability from hormetic doses, small particle sizes, and dynamic adaptive interactions. Dose-Response 12:202–232CrossRefGoogle Scholar
  8. Benelli G (2018) Gold nanoparticles - against parasites and insect vectors. Acta Trop 178:73–80CrossRefGoogle Scholar
  9. Bialek R, Binder N, Dietz K, Joachim A, Knobloch J, Zelck UE (2002) Comparison of fluorescence, antigen and PCR assays to detect Cryptosporidium parvum in faecal specimens. Diagn Microbiol Infect Dis 43:283–288CrossRefGoogle Scholar
  10. Cacciò SM, Chalmers RM (2016) Human cryptosporidiosis in Europe. Clin Microbiol Infect 22:471–480CrossRefGoogle Scholar
  11. Calvo P, Remuñán-López C, Vila-Jato J, Alonso MJ (1997) Novel hydrophilic chitosan – polyethylene oxide nanoparticles as protein carriers. J Appl Polym Sci 63:125–132CrossRefGoogle Scholar
  12. Cameron P, Gaiser BK, Bhandari B, Bartley PM, Katzer F, Bridle H (2016) Silver nanoparticles decrease the viability of Cryptosporidium parvum oocysts. Appl Environ Microbiol 82:431–437CrossRefGoogle Scholar
  13. Castro-Hermida J, Porsi I, Ares-Mazas E, Chartier C (2004) In vitro activity on Cryptosporidium parvum oocyst of different drugs with recognized anti-cryptosporidial efficacy. Rev Med Vet (Toulouse) 155:453–456Google Scholar
  14. Centers for Disease Control and Prevention CDC (2016) Hyperchlorination to kill Cryptosporidium when chlorine stabilizer 1 is NOT in water. USGoogle Scholar
  15. Chaubey P, Mishra B (2014) Mannose-conjugated chitosan nanoparticles loaded with rifampicin for the treatment of visceral leishmaniasis. Carbohydr Polym 101:1101–1108CrossRefGoogle Scholar
  16. Checkley W, White AC, Jaganath D, Arrowood MJ, Chalmers RM, Chen X-M, Fayer R, Griffiths JK, Guerrant RL, Hedstrom L, Huston CD, Kotloff KL, Kang G, Mead JR, Miller M, Petri WA, Priest JW, Roos DS, Striepen B, Thompson RCA, Ward HD, Van Voorhis WA, Xiao L, Zhu G, Houpt ER (2015) A review of the global burden, novel diagnostics, therapeutics, and vaccine targets for Cryptosporidium. Lancet Infect Dis 15:85–94CrossRefGoogle Scholar
  17. Efstratiou A, Ongerth JE, Karanis P (2017) Waterborne transmission of protozoan parasites: review of worldwide outbreaks - an update 2011–2016. Water Res 114:14–22CrossRefGoogle Scholar
  18. Etewa SE, El-Maaty DAA, Hamza RS, Metwaly AS, Sarhan MH, Abdel-Rahman SA, Fathy GM, El-Shafey MA (2018) Assessment of spiramycin-loaded chitosan nanoparticles treatment on acute and chronic toxoplasmosis in mice. J Parasit Dis 42:102–113CrossRefGoogle Scholar
  19. Fernández-Urrusuno R, Calvo P, Remuñán-López C, Vila-Jato JL, Alonso MJ (1999) Enhancement of nasal absorption of insulin using chitosan nanoparticles. Pharm Res 16:1576–1581CrossRefGoogle Scholar
  20. Finch GR, Daniels CW, Black EK, Schaefer FW 3rd, Belosevic M (1993) Dose response of Cryptosporidium parvum in outbred neonatal CD-1 mice. Pharm Res 16:1576–1581Google Scholar
  21. Gaafar MR, Mady RF, Diab RG, Shalaby TI (2014) Chitosan and silver nanoparticles: promising anti-Toxoplasma agents. Exp Parasitol 143:30–38CrossRefGoogle Scholar
  22. Grenha A, Seijo B, Remuñán-López C (2005) Microencapsulated chitosan nanoparticles for lung protein delivery. Eur J Pharm Sci 25:427–437CrossRefGoogle Scholar
  23. Gong C, Cao XF, Deng L, Li W, Huang X-M, Lan J-C, Xiao Q-C, Zhong ZJ, Feng F, Zhang Y, Wang WB, Guo P, Wu K-Y, Peng GN (2017) Epidemiology of Cryptosporidium infection in cattle in China: a review. Parasite 24:1CrossRefGoogle Scholar
  24. Henriksen SA, Pohlenz JF (1981) Staining of cryptosporidia by a modified Ziehl-Neelsen technique. Acta Vet Scand 22:594–296Google Scholar
  25. Holubová N, Zikmundová V, Limpouchová Z, Sak B, Konečný R, Hlásková L, Rajský D, Kopacz Z, McEvoy J, Kváč M (2019) Cryptosporidium proventriculi sp. n. (Apicomplexa: Cryptosporidiidae) in Psittaciformes birds. Eur J Protistol 69:70–87CrossRefGoogle Scholar
  26. Hussein EM, Ahmed SA, Mokhtar AB, Elzagawy SM, Yahi SH, Hussein AM, El-Tantawey F (2018) Antiprotozoal activity of magnesium oxide (MgO) nanoparticles against Cyclospora cayetanensis oocysts. Parasitol Int 67:666–674CrossRefGoogle Scholar
  27. Jain KK (2008) Nanomedicine: application of nanobiotechnology in medical practice. Med Princ Pract 17:89–101CrossRefGoogle Scholar
  28. Jamil B, Habib H, Abbasi S, Nasir H, Rahman A, Rehman A, Bokhari H, Imran M (2016) Cefazolin loaded chitosan nanoparticles to cure multi drug resistant gram-negative pathogens. Carbohydr Polym 136:682–691CrossRefGoogle Scholar
  29. Kao TC, Ungar BLP (1994) Comparison of sequential, random, and haemocytometer methods for counting Cryptosporidium oocysts. J Parasitol 80:816–819CrossRefGoogle Scholar
  30. Karanis P, Kourenti C, Smith H (2007) Water-borne transmission of protozoan parasites: a worldwide review of outbreaks and lessons learnt. J Water Health 5:1–38CrossRefGoogle Scholar
  31. Karanis P, Schoenen D (2001) Biological test for the detection of low concentrations of infectious Cryptosporidium parvum oocysts in water. Acta Hydrochim Hydrobiol 29:242–245CrossRefGoogle Scholar
  32. Katas H, Raja MAG, Lam KL (2013) Development of chitosan nanoparticles as a stable drug delivery system for protein/siRNA. Int J Biomater 2013:146320CrossRefGoogle Scholar
  33. Kayser O (2001) A new approach for targeting to Cryptosporidium parvum using mucoadhesive nanosuspensions: research and applications. Int J Pharm 214:83–85CrossRefGoogle Scholar
  34. Koehler AV, Korhonen PK, Hall RS, Young ND, Wang T, Haydon SR, Gasser RB (2017) Use of a bioinformatic-assisted primer design strategy to establish a new nested PCR-based method for Cryptosporidium. Parasit Vectors 10:509CrossRefGoogle Scholar
  35. Kong M, Chen XG, Xing K, Park HJ (2010) Antimicrobial properties of chitosan and mode of action: a state of the art review. Int J Food Microbiol 144:51–63CrossRefGoogle Scholar
  36. Korich DG, Marshall MM, Smith HV, O'Grady J, Bukhari Z, Fricker CR, Rosen JP, Clancy JL (2000) Inter-laboratory comparison of the CD-1 neonatal mouse logistic dose-response model for Cryptosporidium parvum oocysts. J Eukaryot Microbiol 47:294–298CrossRefGoogle Scholar
  37. Kotloff KL, Blackwelder WC, Nasrin D, Nataro JP, Farag TH, van Eijk A, Adegbola RA, Alonso PL, Breiman RF, Faruque AS, Saha D, Sow SO, Sur D, Zaidi AK, Biswas K, Panchalingam S, Clemens JD, Cohen D, Glass RI, Mintz ED, Sommerfelt H, Levine MM (2012) The Global Enteric Multicenter Study (GEMS) of diarrheal disease in infants and young children in developing countries: epidemiologic and clinical methods of the case/control study. Clin Infect Dis 55(Suppl 4):S232–S245CrossRefGoogle Scholar
  38. Kourenti C, Karanis P (2006) Evaluation and applicability of a purification method coupled with nested PCR for the detection of Toxoplasma oocysts in water. Lett Appl Microbiol 43:475–481CrossRefGoogle Scholar
  39. Lee EH, Khan I, Oh D-H (2018) Evaluation of the efficacy of nisin-loaded chitosan nanoparticles against foodborne pathogens in orange juice. J Food Sci Technol 55:1127–1133CrossRefGoogle Scholar
  40. Ma P, Soave R (1983) Three-step stool examination for cryptosporidiosis in 10 homosexual men with protracted watery diarrhoea. J Infect Dis 147:824–828CrossRefGoogle Scholar
  41. Mammeri M, Chevillot A, Thomas M, Polack B, Julien C, Marden JP, Auclair E, Vallée I, Adjou KT (2018) Efficacy of chitosan, a natural polysaccharide, against Cryptosporidium parvum in vitro and in vivo in neonatal mice. Exp Parasitol 194:1–8CrossRefGoogle Scholar
  42. Marei N, Elwahy AHM, Salah TA, El Sherif Y, El-Samie EA (2019) Enhanced antibacterial activity of Egyptian local insects’ chitosan-based nanoparticles loaded with ciprofloxacin-HCl. Int J Biol Macromol 126:262–272CrossRefGoogle Scholar
  43. Mohammed M, Syeda J, Wasan K, Wasan E (2017) An overview of chitosan nanoparticles and its application in non-parenteral drug delivery. Pharmac 9:E53Google Scholar
  44. Muzzarelli RAA, Boudrant J, Meyer D, Manno N, DeMarchis M, Paoletti MG (2012) Current views on fungal chitin/chitosan, human chitinases, food preservation, glucans, pectins and inulin: a tribute to Henri Braconnot, precursor of the carbohydrate polymers science, on the chitin bicentennial. Carbohydr Polym 87:995–1012CrossRefGoogle Scholar
  45. Nagpal K, Singh SK, Mishra DN (2010) Chitosan nanoparticles: a promising system in novel drug delivery. Chem Pharm Bull (Tokyo) 58:1423–1430CrossRefGoogle Scholar
  46. Nehra P, Chauhan R, Garg N, Verma K (2018) Antibacterial and antifungal activity of chitosan coated iron oxide nanoparticles. Br J Biomed Sci 75:13–18CrossRefGoogle Scholar
  47. Omarova A, Tussupova K, Berndtsson R, Kalishev M, Sharapatova K (2018) Protozoan parasites in drinking water: a system approach for improved water, sanitation and hygiene in developing countries. Int J Environ Res Public Health 15:495CrossRefGoogle Scholar
  48. Plutzer J, Karanis P (2009) Genetic polymorphism in Cryptosporidium species: an update. Vet Parasitol 165:187–199CrossRefGoogle Scholar
  49. Potdar PD, Shetti AU (2016) Evaluation of anti-metastatic effect of chitosan nanoparticles on oesophageal cancer-associated fibroblasts. J Cancer Metastasis Treat 2:259–267CrossRefGoogle Scholar
  50. Qi L, Xu Z, Jiang X, Hu C, Zou X (2004) Preparation and antibacterial activity of chitosan nanoparticles. Carbohydr Res 339:2693–2700CrossRefGoogle Scholar
  51. Ryan U, Hijjawi N, Xiao L (2018) Foodborne cryptosporidiosis. Int J Parasitol 48:1–12CrossRefGoogle Scholar
  52. Robertson LJ, Gjerde BK (2007) Cryptosporidium oocysts: challenging adversaries? Trends Parasitol 23:344–347CrossRefGoogle Scholar
  53. Said DE, ElSamad LM, Gohar YM (2012) Validity of silver, chitosan, and curcumin nanoparticles as anti-Giardia agents. Parasitol Res 111:545–554CrossRefGoogle Scholar
  54. Searcy KE, Packman AI, Atwill ER, Harter T (2006) Capture and retention of Cryptosporidium parvum oocysts by Pseudomonas aeruginosa biofilms. Appl Environ Microbiol 72:6242–6247CrossRefGoogle Scholar
  55. Shahiduzzaman M, Daugschies A (2012) Therapy and prevention of cryptosporidiosis in animals. Vet Parasitol 188:203–214CrossRefGoogle Scholar
  56. Shetta A, Kegere J, Mamdouh W (2019) Comparative study of encapsulated peppermint and green tea essential oils in chitosan nanoparticles: encapsulation, thermal stability, in-vitro release, antioxidant and antibacterial activities. Int J Biol Macromol 126:731–742CrossRefGoogle Scholar
  57. Teimouri A, Azami SJ, Keshavarz H, Esmaeili F, Alimi R, Mavi SA, Shojaee S (2018) Anti-Toxoplasma activity of various molecular weights and concentrations of chitosan nanoparticles on tachyzoites of RH strain. Int J Nanomedicine 13:1341–1351CrossRefGoogle Scholar
  58. Tripathy S, Das S, Chakraborty SP, Sahu SK, Pramanik P, Roy S (2012) Synthesis, characterization of chitosan–tripolyphosphate conjugated chloroquine nanoparticle and its in vivo anti-malarial efficacy against rodent parasite: a dose and duration dependent approach. Int J Pharm 434:292–305CrossRefGoogle Scholar
  59. Unciti-Broceta JD, Arias JL, Maceira J, Soriano M, Ortiz-González M, Hernández-Quero J, Muñóz-Torres M, de Koning HP, Magez S, Garcia-Salcedo JA (2015) Specific cell targeting therapy bypasses drug resistance mechanisms in African trypanosomiasis. PLoS Pathog 11:e1004942CrossRefGoogle Scholar
  60. Ungar BL, Burris JA, Quinn CA, Finkelman FD (1990) New mouse models for chronic Cryptosporidium infection in immunodeficient hosts. Infect Immun 58:961–969Google Scholar
  61. Vaezifar S, Razavi S, Golozar MA, Karbasi S, Morshed M, Kamali M (2013) Effects of some parameters on particle size distribution of chitosan nanoparticles prepared by ionic gelation method. J Clust Sci 24:891–903CrossRefGoogle Scholar
  62. Villanueva MT (2017) Infectious diseases: decrypting Cryptosporidium. Nat Rev Drug Discov 16:527–527CrossRefGoogle Scholar
  63. Wang J, Zeng ZW, Xiao RZ, Xie T, Zhou GL, Zhan XR, Wang SL (2011) Recent advances of chitosan nanoparticles as drug carriers. Int J Nanomedicine 6:765–774Google Scholar
  64. Xing K, Chen XG, Liu CS, Cha DS, Park HJ (2009) Oleoyl-chitosan nanoparticles inhibits Escherichia coli and Staphylococcus aureus by damaging the cell membrane and putative binding to extracellular or intracellular targets. Int J Food Microbiol 132:127–133CrossRefGoogle Scholar
  65. Yong SK, Shrivastava M, Srivastava P, Kunhikrishnan A, Bolan N (2015) Environmental applications of chitosan and its derivatives. Rev Environ Contam Toxicol 233:1–43Google Scholar

Copyright information

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

Authors and Affiliations

  • Shahira A. Ahmed
    • 1
    Email author
  • Heba S. El-Mahallawy
    • 2
  • Panagiotis Karanis
    • 3
  1. 1.Department of Parasitology, Faculty of MedicineSuez Canal UniversityIsmailiaEgypt
  2. 2.Department of Animal Hygiene, Zoonoses and Animal Behaviour and Management, Faculty of Veterinary MedicineSuez Canal UniversityIsmailiaEgypt
  3. 3.University of CologneMedical Faculty and University HospitalCologneGermany

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