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Angiostrongylus cantonensis: An optimized cultivation of this parasitic nematode under laboratory conditions

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Abstract

Angiostrongylus cantonensis (A. cantonensis), a parasitic nematode, is the important neurotropic pathogen which causes human angiostrongyliasis. It has a complex life-cycle and severe parasite-host interaction in contrast to free-living nematode. Establishment of a well-suited life-cycle and in vitro cultivation of A. cantonensis in the laboratory will be one of the key techniques to elucidate the mechanism of parasite-host interaction. However, the low survival and growth rate of worms is still to be the problem. We optimized the known life-cycle of A. cantonensis in the laboratory, showing that small in size, easy to breed, and high compatibility of Biomphalaria straminea precede the common snails as an intermediate host of A. cantonensis. Furthermore, the egg hatching rate in Ham’s F-12 medium reached approximately 80% using the eggs of mature female adult worms. We also demonstrated that the survival of larvae could be sustained for more than 30 days by in vitro cultivation of L1 larvae in DMEM with mixed antibiotics (100 units/mL of penicillin G potassium, 50 μg/mL of streptomycin sulfate, and 0.5 μg/mL of amphotericin B) and L3, L4, and L5 larvae in Waymouth’s medium with 20% fetal calf serum and mixed antibiotics. Infective L1 and L3 larvae kept high infective rate to the snail and rat after cultivation in these media, respectively. It will provide the basis for studying on genetic manipulations for functional genes, new drug screening, and the mechanism of parasite-host interaction of parasitic nematodes.

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Abbreviations

A. cantonensis :

Angiostrongylus cantonensis

A. costaricensis :

Angiostrongylus costaricensis

A. fulica :

Achatina fulica

C. elegans :

Caenorhabditis elegans

C. hamulosa :

Cheilospirura hamulosa

References

  • Bethony J, Brooker S, Albonico M, Geiger SM, Loukas A, Diemert D, Hotez PJ (2006) Soil-transmitted helminth infections: ascariasis, trichuriasis, and hookworm. Lancet 367(9521):1521–1532

    Article  PubMed  Google Scholar 

  • Bhaibulaya M (1975) Comparative studies on the life history of Angiostrongylus mackerrasae Bhaibulaya, 1968 and Angiostrongylus cantonensis (Chen, 1935). Int J Parasitol 5(1):7–20

    Article  CAS  PubMed  Google Scholar 

  • Blaxter M (1998) Caenorhabditis elegans is a nematode. Science 282(5396):2041–2046

    Article  CAS  PubMed  Google Scholar 

  • Ghedin E, Wang S, Spiro D, Caler E, Zhao Q, Crabtree J, Allen JE, Delcher AL, Guiliano DB, Miranda-Saavedra D, Angiuoli SV, Creasy T, Amedeo P, Haas B, El-Sayed NM, Wortman JR, Feldblyum T, Tallon L, Schatz M, Shumway M, Koo H, Salzberg SL, Schobel S, Pertea M, Pop M, White O, Barton GJ, Carlow CK, Crawford MJ, Daub J, Dimmic MW, Estes CF, Foster JM, Ganatra M, Gregory WF, Johnson NM, Jin J, Komuniecki R, Korf I, Kumar S, Laney S, Li BW, Li W, Lindblom TH, Lustigman S, Ma D, Maina CV, Martin DM, McCarter JP, McReynolds L, Mitreva M, Nutman TB, Parkinson J, Peregrín-Alvarez JM, Poole C, Ren Q, Saunders L, Sluder AE, Smith K, Stanke M, Unnasch TR, Ware J, Wei AD, Weil G, Williams DJ, Zhang Y, Williams SA, Fraser-Liggett C, Slatko B, Blaxter ML, Scott AL (2007) Draft genome of the filarial nematode parasite Brugia malayi. Science 317(5845):1756–1760

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Gu J, Liu M, Li H, Luo Y, Li X, Chen X (2008) Construction of the life cycle of Angiostrongylus cantonensis in laboratory. J South Med Univ 28(4):551–554

    Google Scholar 

  • Hata H (1993) In vitro cultivation of the third and fourth stage larvae of Angiostrongylus cantonensis. J Vet Med Sci 55(55):345–347

    Article  CAS  PubMed  Google Scholar 

  • Hata H (1996) In vitro cultivation of Angiostrongylus costaricensis eggs to first stage larvae in chemically defined medium. Int J Parasitol 26(3):281–286

    Article  CAS  PubMed  Google Scholar 

  • Hata H, Kojima S (1990) Angiostrongylus cantonensis: in vitro cultivation from the first-stage to infective third-stage larvae. Exp Parasitol 70(4):476–482

    Article  CAS  PubMed  Google Scholar 

  • Hata H, Kojima S (1991) Angiostrongylus costaricensis: culture of third-stage larvae to young adults in a defined medium. Exp Parasitol 73(3):354–361

    Article  CAS  PubMed  Google Scholar 

  • Kim JR, Hayes KA, Yeung NW, Cowie RH (2014) Diverse gastropod hosts of Angiostrongylus cantonensis, the rat lungworm, globally and with a focus on the Hawaiian Islands. PLoS One 9(5):e94969

    Article  PubMed  PubMed Central  Google Scholar 

  • Lin RJ, He JW, Chung LY, Lee JD, Wang JJ, Yen CM (2013) Angiostrongylus cantonensis (Nematode: Metastrongiloidea): in vitro cultivation of infective third-stage larvae to fourth-stage larvae. PLoS One 8(8):1–10

    Google Scholar 

  • Liu Y, Gao ST, Geng YJ (2007) Experimental study on the infectiveness of Angiostrongylus cantonensis to three species of freshwater snails. China Trop Med 7(9):1517–1519

    Google Scholar 

  • Liu HX, Zhang Y, Lv S, Hu L, Zhou XN (2009) Establishment and observation of the life cycle of Angiostrongylus cantonensis in a laboratory setting. J Pathog Biol: 836–839

  • Luessi F, Sollors J, Torzewski M, Müller HD, Siegel E, Blum J, Sommer C, Vogt T, Thömke F (2009) Eosinophilic meningitis due to Angiostrongylus cantonensis in Germany. J Travel Med 64(12):351–352

    Google Scholar 

  • Mackerras MJ, Sandars DF (1954) Life history of the rat lung-worm and its migration through the brain of its host. Nature 173(4411):956–957

    Article  CAS  PubMed  Google Scholar 

  • Martins YC, Tanowitz HB, Kazacos KR (2015) Central nervous system manifestations of Angiostrongylus cantonensis infection. Acta Trop 141(Pt A):46–53

    Article  PubMed  Google Scholar 

  • Morassutti AL, Perelygin A, DE Carvalho MO, Lemos LN, Pinto PM, Frace M, Wilkins PP, Graeff-Teixeira C, DA Silva AJ (2013) High throughput sequencing of the Angiostrongylus cantonensis genome: a parasite spreading worldwide. Parasitology 140(10):1304–1309

    Article  PubMed  Google Scholar 

  • Park SY, Fox LM (2013) Angiostrongylus cantonensis: epidemiology in the continental United States and Hawa'i. Hawaii J Med Public Health 72(6 Suppl 2)

  • Pullan RL, Smith JL, Jasrasaria R, Brooker SJ (2014) Global numbers of infection and disease burden of soil transmitted helminth infections in 2010. Parasite Vector 7:37

    Article  Google Scholar 

  • Richards CS, Merritt JW (1967) Studies on Angiostrongylus cantonensis in molluscan intermediate hosts. J Parasitol 53(53):382–388

    Article  CAS  PubMed  Google Scholar 

  • Robles Mdel R, Navone GT, Kinsella JM (2008) A new angiostrongylid (Nematoda) species from the pulmonary arteries of Akodon azarae (Rodentia: Cricetidae) in Argentina. J Parasitol 94(2):515–519

    Article  PubMed  Google Scholar 

  • Simoes RO, Monteiro FA, Sanchez E, Thiengo SC, Garcia JS, Costa-Neto SF, Luque JL, Maldonado A (2011) Endemic angiostrongyliasis, Rio de Janeiro, Brazil. Emerg Infect Dis 17(7):1331–1333

    Article  PubMed  PubMed Central  Google Scholar 

  • Taylor MJ, Hoerauf A, Bockarie M (2010) Lymphatic filariasis and onchocerciasis. Lancet 376(9747):1175–1185

    Article  PubMed  Google Scholar 

  • Thiengo SC, Maldonado A, Mota EM, Torres EJ, Caldeira R, Carvalho OS, Oliveira AP, Simões RO, Fernandez MA, Lanfredi RM (2010) The giant African snail Achatina fulica as natural intermediate host of Angiostrongylus cantonensis in Pernambuco, northeast Brazil. Acta Trop 115(3):194–199

    Article  CAS  PubMed  Google Scholar 

  • Thiengo SC, Simões RDO, Fernandez MA, Maldonado Júnior A, Cowie RH (2013) Angiostrongylus cantonensis and rat lungworm disease in Brazil. Hawaii J Med Public Health 72(6 Suppl 2):18–22

    PubMed  PubMed Central  Google Scholar 

  • Uga S, Matsumura T (1982) In vitro cultivation of Angiostrongylus cantonensis eggs. Jpn J Parasitol 31:59–66

    Google Scholar 

  • Wang QP, Lai DH, Zhu XQ, Chen XG, Lun ZR (2008) Human angiostrongyliasis. Lancet Infect Dis 8(10):621–630

    Article  PubMed  Google Scholar 

  • Wang QP, Wu ZD, Wei J, Owen RL, Lun ZR (2012) Human Angiostrongylus cantonensis: an update. Eur J Clin Microbiol Infect Dis 31(4):389–395

    Article  PubMed  Google Scholar 

  • World Health Organization (2015) Investing to overcome the global impact of neglected tropical diseases. Wkly Epidemiol Rec 1(2):91–92

    Google Scholar 

  • Yu S, Tang LH, Zhou SS, Chen YD, Yang YC, Lin SX (2010) Stunting and soil-transmitted-helminth infections among school-age pupils in rural areas of southern China. Parasite Vector 3(1):97

    Article  Google Scholar 

  • Zhang Z (2013) Animal biodiversity: an update of classification and diversity in 2013. In: Zhang ZQ (ed) Animal biodiversity: an outline of higher-level classification and survey of taxonomic richness (Addenda 2013) Zootaxa 3703:1–82

  • Zhou W, Yu S, Chen D, Lin J, Guo Y, Chen S (2007) Description on the intermediate hosts of Angiostrongylus cantonensis. Chinese J Zoonoses 23(04):401–408

    Google Scholar 

Download references

Acknowledgement

This work was supported by the National Key Research and Development Plan of China (2016YFC1200500 and 2016YFC1202003), the National Natural Science Foundation of China (81401686), and the PhD Start-up Fund of the Natural Science Foundation of Guangdong Province, China (2015A030310058).

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Authors and Affiliations

  1. Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, No. 74, Zhongshan Rd. 2, Guangzhou, 510080, People’s Republic of China

    Hui Xie, Dongjuan Yuan, Shiqi Luo, Xingda Zeng, Xin Zeng, Ping He, Zhiyue Lv & Zhongdao Wu

  2. Key Laboratory for Tropical Diseases Control (Sun Yat-sen University), The National Ministry of Education, Guangzhou, 510080, People’s Republic of China

    Hui Xie, Dongjuan Yuan, Shiqi Luo, Xingda Zeng, Xin Zeng, Ping He, Zhiyue Lv & Zhongdao Wu

  3. Provincial Engineering Technology Research Center for Diseases-Vectors Control, Guangzhou, 510080, Guangdong, People’s Republic of China

    Dongjuan Yuan, Zhiyue Lv & Zhongdao Wu

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  1. Hui Xie
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  2. Dongjuan Yuan
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  3. Shiqi Luo
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  5. Xin Zeng
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  7. Zhiyue Lv
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Correspondence to Dongjuan Yuan or Zhongdao Wu.

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The authors declare that they have no competing interests.

Ethics statement

All procedures of the animals were carried out according to the guidelines of the Association for Assessment and Accreditation of Laboratory Animal Care International. Our protocols followed institutional ethical guidelines that were approved by the Ethics Committee of ZSSOM on Laboratory Animal Care (SYSU, ZSSOM; Permit No: 2016-122).

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Xie, H., Yuan, D., Luo, S. et al. Angiostrongylus cantonensis: An optimized cultivation of this parasitic nematode under laboratory conditions. Parasitol Res 116, 2231–2237 (2017). https://doi.org/10.1007/s00436-017-5526-7

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  • DOI: https://doi.org/10.1007/s00436-017-5526-7

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