Abstract
A cryopreservation protocol, with an optimum survival rate of 86.8 %, was developed for a strain of Steinernema carpocapsae (Weiser) ItS-CAO1 isolated in a lagoon plain in Veneto Region (northern Italy). Some laboratory bioassays were carried out against Galleria mellonella (L.) (Lepidoptera: Pyralidae) larvae to determine the infectivity and stability of this strain after cryopreservation. Insect mortality was very high in different assays (sand column and penetration assays), while the percentage of penetrating infective juveniles was 59.4 and 33.8 respectively. Larval mortality in a one-on-one quality assay was 66.6 % while in an exposure time assay the exposure time to nematodes required to achieve 50 % insect mortality was lower than 20 min. With the results of infectivity assays we can evaluate the possibility to use cryopreservation for long-term storage of entomopathogenic nematodes for biological control programs.
Similar content being viewed by others
References
Bai C, Shapiro-Ilan DI, Gaugler R, Shuxia Y (2004) Effect of entomopathogenic nematode concentration on survival during cryopreservation in liquid nitrogen. Journal of Nematology 36(3):281–284
Bai C, Shapiro-Ilan DI, Gaugler R, Hopper KR (2005) Stabilization of beneficial traits in Heterorhabditis bacteriophora through creation of inbred lines. Biol Control 32:220–227
Bednarek A, Nowicki T, Wojcik WF (1986) Sex structure in populations of Steinernema feltiae. Zesz. Probl. Postepow Nauk Roln. 323:213–223
Begley JW (1990) Efficacy against insects in habitats other than soil. In: Gaugler R, Kaya HK (eds) Entomopathogenic nematodes in biological control. CRC Press, Boca Raton, pp 215–231
Behm C (1997) The role of trehalose in the physiology of nematodes. Int J Parasitol 27:215–229
Bilgrami AL, Gaugler R, Shapiro-Ilan DI, Adams BJ (2006) Source of trait deterioration in entomopathogenic nematodes Heterorhabditis bacteriophora and Steinernema carpocapsae during in vivo culture. Nematology 8(3):397–409
Boemare N (2002) Biology, taxonomy and systematic of Phothorhabdus and Xenorhabdus. In: Gaugler R (ed) Entomopathogenic nematology. CAB International, Wallingford, pp 35–56
Campos-Herrera R, Gutierrez C (2009) A laboratory study on the activity of Steinernema feltiae (Rhabditida: Steinernematidae) Rioja strain against horticultural insect pests. J Pest Sci 82:305–309
Caroli L, Glazer I, Gaugler R (1996) Entomopathogenic nematode infectivity assay: multi variable comparison of penetration into different hosts. Biocontrol Sci Tech 6:227–233
Cosi E, Triggiani O, Irdani T, Tarasco E, Roversi PF (2008) First results of entomopathogenic nematodes cryopreservation in liquid nitrogen and storage at −140°C. REDIA XCI:181–183
Curran JC, Gilbert C, Butler K (1992) Routine cryopreservation of isolates of Steinernema and Heterorhabditis spp. Journal of Nematology 24:269–270
Gaugler R, Kaya HK (1990) Entomopathogenic nematodes in biological control. CRC Press Inc, Boca Raton
Glazer I (1991) Invasion rate as a measure of infestivity of steinernematid and heterorhabditid nematodes to insects. J Invertebr Pathol 59:90–94
Glazer I, Lewis EE (2000) Bioassays for entomopathogenic nematodes. In: Navon A, Ascher KRS (eds) Bioassays of entomopathogenic microbes and nematodes. CABI Publishing, Wallingford, pp 229–247
Grewal PS, Selvan S, Lewis EE, Gaugler R (1993) Male insect-parasitic nematodes: a colonizing sex. Experientia 49:605–608
Griffin CT, Downes MJ (1994) Recognition of low temperature active isolates of the entomopathogenic nematode Heterorhabditis. Nematologica 37:83–91
Irdani T, Carletti B, Ambrogioni L, Roversi PF (2006) Rapid-cooling and storage of plant nematodes at −140 °C. Cryobiology 52:319–322
Irdani T, Scotto C, Roversi PF (2011) Low cryoprotectant concentrations and fast cooling for nematode cryostorage. Cryobiology 63:12–16
Jagdale GB, Grewal PS (2003) Acclimation of entomopathogenic nematodes to novel temperatures: Trehalose accumulation and the acquisition of thermotolerance. Int J Parasitol 33:145–152
Kaya HK, Stock SP (1997) Techniques in insect nematology. In: Lacey LA (ed) Techniques in insect pathology. Academic Press, London, pp 281–324
Klein MG (1990) Efficacy against soil-inhabiting insect pests. In: Gaugler R, Kaya H (eds) Entomopathogenic nematodes in biological control. CRC Press, Boca Raton, pp 195–214
Leopold RA (1998) Cold storage of insects for integrated pest management. In: Hallman GJ, Dellinger DL (eds) Temperature sensitivity in insects and application in integrated pest management. Westview Press, Boulder, pp 235–267
Mazur P (1985) Basic concepts in freezing cells. Proceeding of the first International Conference on Deep freezing of Boar Semen, Uppsala, pp 99–111
Mazur P, Schneider U, Mahowald AP (1992) Characteristics and kinetics of subzero chilling injury in Drosophila embryos. Cryobiology 29:39–68
Miller RW (1989) Novel pathogenicity assessment technique for Steinernema and Heterorhabditis entomopathogenic nematodes. Journal of Nematology 21:574 (Abstr.)
Nugent MJ, O’Leary SA, Burnell AM (1996) Optimised procedures for the cryopreservation of different species of Heterorhabditis. Fundamental and Applied Nematology 19:1–6
Poinar GO Jr (1990) Biology and taxonomy of Steinernematidae and Heterorhabditidae. In: Gaugler R, Kaya HK (eds) Entomopathogenic nematodes in biological control. CRC Press, Boca Raton, pp 23–61
Popiel I, Vasquez EM (1991) Cryopreservation of Steinernema carpocapsae and Heterorhabditis bacteriophora. Journal of Nematology 21:432–437
Qiu L, Bedding RA (1999) Low temperature induced cryoprotectants synthesis by the infective juveniles of Steinernema carpocapsae: its biological significance and the mechanism involved. Cryo letters 20(6):393–404
Roversi PF, Cosi E, Irdani T (2008) Chill sensitivity and cryopreservation of eggs of the greater wax moth Galleria mellonella (Lepidoptera: Pyralidae). Cryobiology 56:1–7
Sayre RM, Hwang SW (1975) Freezing and storing Ditylenchus dipsaci in liquid nitrogen. Journal of Nematology 7:199–202
Shapiro DI, Glazer I, Segal D (1996) Trait stability and fitness of the heat tolerant entomopathogenic nematode Heterorhabditis bacteriophora IS5 strain. Biol Control 6:238–244
Stock SP, Pryor BM, Kaya HK (1999) Distribution of entomopathogenic nematodes (Steinernematidae and Heterorhabditidae) in natural habitats in California, USA. Biodivers Conserv 8:535–549
Torrini G, Landi S, Benvenuti C, De Luca F, Fanelli E, Troccoli A, Tarasco E, Bazzoffi P, Roversi PF (2014) Morphological and molecular characterization of a Steinernema carpocapsae (Nematoda Steinernematidae) strain isolated in Veneto region (Italy). REDIA XCVII:89–94
van der Beek HJG, Veldhuis WBJ, Zijlstra C, van Silfhout CH (1996) Preservation of Meloidogyne hapla and M. chitwoodi in liquid nitrogen: Differences in response between populations. Fundamental and Applied Nematology 19:227–234
Wang XD, Grewal PS (2002) Rapid genetic deterioration of environmental tolerance and reproductive potential of an entomopathogenic nematode during laboratory maintenance. Biol Control 23:71–78
Acknowledgments
Thanks to Peter Christie for the English revision.
Author information
Authors and Affiliations
Corresponding author
Additional information
Handling Editor: Ralf Elhers.
Rights and permissions
About this article
Cite this article
Torrini, G., Landi, S., Tarasco, E. et al. Evaluation of Steinernema carpocapsae survival and infectivity after cryopreservation. BioControl 61, 461–469 (2016). https://doi.org/10.1007/s10526-016-9724-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10526-016-9724-5