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Effects of Two Carbamates on Infective Juveniles of Steinernema Carpocapsae All Strain and Steinernema Feltiae Umefi Strain

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Effects of Two Carbamates on Infective Juveniles of Steinernema Carpocapsae All Strain and Steinernema Feltiae Umefi Strain Journal of Nematology 28(3):310-317. 1996. © The Society of Nematologists 1996. Effects of Two Carbamates on Infective Juveniles of Steinernema carpocapsae All Strain and Steinernema feltiae Umefi Strain R. GORDON, J. CHIPPETT, AND J. TILLEY 2 Abstract: Laboratory bioassays were conducted to determine the effects of two carbamates, carbo- furan (an acetylcholinesterase inhibitor) and fenoxycarb (a juvenile hormone analog), on survival and infectivity of the infective juveniles (IJ) of Steinernema feltiae Ume~ strain and Steinernema car- pocapsae All strain. Both insecticides caused mortality of IJ in a dose-related fashion. The two nematode species were equally sensitive to fenoxycarb (LDs0 ca. 0.03mg/ml). Whereas IJ of S. feltiae were several orders of magnitude more sensitive to carbofuran (LDs0 ~ 0.2 ~g/ml) than to fenox- ycarb, S. carpocapsae IJ displayed approximately the same degree of sensitivity to carbofuran (LDs0 0.01-0.03 mg/ml) as they did toward fenoxycarb. Toxicity of the carbamates was the same at all exposure periods from 24 to 168 hours' duration. Determinations of infective doses of nematodes required to cause 50% mortality of Ga~teria mellonella larvae showed that the infectivity of IJ that survived exposure to either of the two carbamates was not compromised by treatment. Key words: acetylcholinesterase inhibitor, carbamate, carbofuran, entomopathogenic nematode, fenoxycarb, infective junvenile, insecticide, juvenile hormone analog, Steinernema carpocapsae, Stein- ernema feltiae. In developing integrated pest manage- ever, the bioassay procedure used in the ment (IPM) strategies involving the use of latter study was incomplete and measured entomopathogenic nematodes (f. Stein- only nematode mobility. Ishibashi (11) re- ernematidae and Heterorhabditidae) and ported that soil applications of carbamate chemical pesticides, it is important to as- and organophosphate insecticides with certain the degree to which these nema- Steinernema carpocapsae All strain actually todes may be affected by the chemicals in- increased the nematode's field efficacy volved. Studies done using chemical insec- against three species of larval Lepidoptera ticides that inhibit acetylcholinesterase by stimulating nictating behaviour and in- activity have yielded inconsistent results, fectivity of the IJ. suggesting that it is not possible to gener- However, several carbamates and orga- alize with respect to insecticide tolerance. nophosphates adversely affected the in In early laboratory studies, infective juve- vitro development and reproduction of S. niles (IJ) of Steinernema carpocapsae were carpocapsae All strain, whereas the nema- unaffected by short-term exposure to a tode was unaffected by the chlorinated hy- wide variety of insecticides that were toxic drocarbon methoxychlor or the synthetic to other soil-dwelling nematode species pyrethroid fenvalerate (9). Infective juve- (4). When exposure time to the insecticides niles of this nematode displayed partial pa- was increased beyond 24 hours, however, ralysis and loss of infectivity consequent to nematode mortality increased (5). Infec- being treated with certain of these insecti- tive juveniles of the DD136 strain of S. car- cides (10,14). In a subsequent study, it was pocapsae were reported to be unaffected by determined that IJ of this strain of S. car- treatment in the laboratory with a variety pocapsae were refractory to a variety of car- of insecticides and fungicides (2). How- bamates but sensitive to certain organo- phosphates and the tertiary amine cartap (26). Laboratory bioassays involving expo- Received for publication 20 December 1995. sure of IJ of an Italian strain of S. carpo- 1 Supported by a grant from the Natural Sciences and En- capsae and an unspecified strain of S. feltiae gineering Research Council of Canada (Research Grant A6679). to several categories of pesticides showed 2 Department of Biology, Memorial University of New- that while most of the herbicides and fun- foundland, St. John's, Newfoundland, Canada A1B 3X9. E-mail: [email protected] gicides were not toxic, a high proportion 310 Carbamates on Steinernematids: Gordon et al. 311 of insecticides, acaricides, and nematicides tained from Biologic Biocontrol Products, induced adverse effects ranging from im- Willow Hill, Pennsylvania, U.S.A. This paired movement and infectivity to death nematode is marketed as S. carpocapsae of the IJ (20). Heterorhabditids were de- Ume~ strain and was first isolated from termined to be more susceptible than soil in Sweden (18). However, we have ob- steinernematids to pesticides (22), with served that its morphometric measure- Heterorhabditis bacteriophora and Heterorhab- ments correspond to those of S. feltiae ditis heliothidis displaying approximately (Jagdale and Gordon, unpublished obser- the same overall intolerance but differing vations). Restriction fragment length poly- in their sensitivities to specific pesticides morphisms of amplified rDNA showed a (21). close association between the Ume~t strain Due to environmental concerns, current and several S. feltiae strains (13). There- trends are toward decreased reliance on fore, we believe that the nematode being conventional chemical insecticides. Juve- marketed as S. carpocapsae Ume~ strain is nile hormone analogs (JHAs) are insecti- improperly named and we have desig- cidal compounds that offer a biorational nated it as S. feltiae Ume~i strain. Both approach to pest management since they nematode isolates were maintained at 25°C specifically target the insect's neuroendo- by propagation through larval waxmoths, crine system and, thus, have minimal del- Galleria mellonella (25). eterious effects on non-target organisms Sources of insecticides: Technical-grade (19). The mermithid nematode Romano- carbofuran was provided by FMC, Prince- mermis culicivorax was unaffected by the ton, New Jersey, U.S.A., and technical- terpenoid methoprene, one of the first grade fenoxycarb by Elanco, Eli Lilly Can- JHAs that was commercially available ada Inc., Scarborough, Ontario, Canada. (6,15). However, the toxicological effects Toxicity of insecticides to IJ : Nematode sus- on entomopathogenic nematodes of JHAs, pensions were prepared by using a dispos- particularly more recently developed non- able Millipore Filter (0.2 m~ pore diam.) to terpenoid ones, are unknown. The present filter off the dilute formaldehyde solution study was done to determine the effects of collected from the White traps (25) in two carbamates on the survival and infec- which IJ had emerged. The IJ that were tivity of IJ of S. carpocapsae All strain and S. trapped on the filter pad were subse- feltiae Ume~ strain. Carbofuran is an ace- quently washed twice by passing distilled tylcholinesterase inhibitor widely used for water through the filter, then resuspend- control of soil-dwelling insects. Fenoxy- ed in distilled water. Using a stereomicro- carb is a recently developed JHA that, ac- scope to count the IJ in 1-ml droplets of cording to the supplier, does not inhibit evenly stirred suspension and adjusting acetylcholinesterase (Technical Data the water volume accordingly, the suspen- Sheet, 6th ed., 1989; Maag Agrochemicals, sion was adjusted to a concentration of 50 Vero Beach, FL). IJ/ml. To allow for better dispersion, insec- ticide solutions were formulated using the emulsifier 0.01% Triton-X-100 (Sigma MATERIALS AND METHODS Chemical Co., St. Louis, MO). Concentra- tions of fenoxycarb were formulated such Sources of nematodes: Steinernema carpocap- that after dilution with the nematode sus- sae All strain was provided by Plant Prod- pension, final concentrations were 0.001, ucts Ltd., Brampton, Ontario, Canada. 0.01, 0.1, 0.5, and 1.0 mg/ml. Final concen- Steinernema feltiae Ume~ strain was pro- trations for carbofuran ranged from vided by R. West, Natural Resources Can- 0.00001 mg/ml to 0.01 mg/ml (S.feltiae) or ada, Canadian Forest Service (CFS), St. from 0.00001 mg/ml to 0.05 mg/ml (S. car- John's, Newfoundland, Canada, from a pocapsae). Equal volumes (10-ml) of nema- stock colony that had been initially ob- tode suspension and insecticide emulsion 312 Journal of Nematology, Volume 28, No. 3, September 1996 were pipetted into glass petri dishes (60 x transferred to filter-paper circles lining 15 ram). Treated and untreated controls the bases of disposable petri dishes (60 x were employed using 0.01% Triton-X- 100 15 mm); it was possible to count the live IJ and distilled water, respectively, in place of as they were removed. Doses of 10, 30, 50, the insecticide emulsion. and 70 IJ were used and distilled water Nematodes were maintained in an incu- (~<0.2 ml) added until the filter papers bator (24°C) in complete darkness. Toxic- were evenly moist. Ten insects were added ity was determined after 24-, 72-, 120-, and to each petri dish, which was closed and 168-hour exposure to insecticide by re- incubated in darkness at 24 °C. Thus, moving 1 ml of suspension containing at doses were 1, 3, 5, and 7 IJ/insect, with five least 25 IJ from the appropriate petri replications of each dose. Insect mortality dishes and probing each nonmotile nema- was determined 5 days after infection; all tode in the sample with a lachrymal needle dead insects were dissected to verify that under a stereomicroscope to determine they harbored steinernematids. whether it was alive or dead (10). This was Statistical analysis: In the IJ toxicity stud- necessary because IJ normally display the ies, the effects of the compounds (% IJ active or inactive behavioral
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