Inhibition of Acetylcholinesterases from Aphelenchus avenae by Carbofuran and Fenamiphos

Exposure to carbofuran and fenamiphos for 72 hours reduced the numbers of active Aphelenchus avenae in aqueous suspension by > 75%. When nematicides were removed, many A. avenae exposed to carbofuran resumed normal movement but A. avenae treated with fenamiphos did not recover. Acetylcholinesterase (AChE) activity was suppressed by > 95% in nematodes treated with carbofuran or fenamiphos. However, 48 hours after treated nematodes had been placed in water, AChE activity in carbofuran treated populations was 98% of the levels in control nematodes. Nematodes that had been treated with fenamiphos showed only slight AChE recovery. The antidotes, atropine sulfate and 2-PAM, were largely ineffective in counteracting the toxic effects of the nematicides.     

Soil Applications of Steinernematid and Heterorhabditid Nematodes for Control of Colorado Potato Beetles,Leptinotarsa decemlineata (Say)

Three strains of Steinernema feltiae Filipjev (All, Mexican, and Breton strains) and one of Heterorhabditis heliothidis (Khan, Brooks, and Hirschmann) were evaluated for their potential to control Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say), larvae and pupae in the soil. In laboratory studies, H. heliothidis and S. feltiae (Mexican strain) produced the highest mortality (6 days posttreatment) of CPB when applied to the surface of a soil column containing mature CPB larvae 5 cm below. Mortality ranged from 80 to 90% at rates of 79-158 nematodes/cm². Similar results were seen in a field microplot study with all four nematodes; S. feltiae (Mexican strain) and H. heliothidis were most effective. Adult CPB emergence was reduced 86.5-100% after application of 31-93 H. heliothidis/cm² and 88.4-100% with 93-155 S. feltiae (Mexican strain)/cm². The All strain of S. feltiae was moderately effective (ca. 80% reduction at 93-155 nematodes/cm²), while the Breton strain was ineffective (< 40% reduction at 155 nematodes/cm²). In small plots of potatoes enclosed in field cages, application of H. heliothidis and S. feltiae (Mexican strain) at rates of 93-155 nematodes/cm² before larval CPB burial in the soil resulted in 66-77% reduction in adult CPB emergence. Soil applications of these nematodes show potential for biological control of CPB.     

Some Factors Affecting Survival of Desiccation by Infective Juveniles of Orrina phyllobia

The survival of desiccation by J4 Orrina phyllobia was examined at controlled relative humidities. When nematodes were transferred from water to air at 10% relative humidity (rh), 80% died within 30 minutes. When nematodes were transferred from water to air with rh at 70% or greater for ca. 15 minutes prior to being transferred to 10% rh, more than 90% of them survived desiccation. This phenomenon is referred to as preconditioning and occurred at much faster rates (2-30 minutes) than has been observed for other nematode species (24 hours). Differences in preconditioning rates may be due to technique-dependent variations in boundary layer resistance around nematodes during desiccation.     

Movement of Five Nematode Species through Sand Subjected to Natural Temperature Gradient Fluctuations

Temperature gradient fluctuations that occur naturally as a result of heating and cooling of the soil surface were reproduced within 15-cm-d, 15-cm-long acrylic tubes filled with moist sand. Sunny and rainy periods during the late summer in eastern Texas were simulated. Five ecologically different nematode species were adapted to fluctuating temperatures for 20-36 hours at a simulated depth of 12.5 cm before being injected simultaneously into the centers of tubes at that depth. When heat waves were propagated horizontally to eliminate gravitational effects, the movement of Ditylenchus phyllobius, Steinernema glaseri, and Heterorhabditis bacteriophora relative to the thermal surface was rapid and largely random. However, Rotylenchulus reniformis moved away from and Meloidogyne incognita moved toward the thermal surface. When heat waves were propagated upward or downward, responses to temperature were the same as when propagated horizontally, irrespective of gravity. The initial direction of movement 1.5 hours after introduction to 20-era-long tubes at five depths at five intervals within a 24-hour cycle indicated that M. incognita moved away from and R. reniformis moved toward the temperature to which last exposed. Differences in movement of the five species tested relative to gravity appeared related to body length, with the smallest nematodes moving downward and the largest moving upward.     

Parasitism of Nematodes by the Fungus Hirsutella rhossiliensis as Affected by Certain Organic Amendments

Experiments were conducted to determine whether the addition of organic matter to soil increased numbers of bacterivorous nematodes and parasitic activity of the nematophagous fungus Hirsutella rhossiliensis. In a peach orchard on loamy sand, parasitism of the plant-parasitic nematode Criconemella xenoplax by H. rhossiliensis was slightly suppressed and numbers of C. xenoplax were not affected by addition of 73 metric tons of composted chicken manure/ha. In the laboratory, numbers of bacterivorous nematodes (especially Acrobeloides spp.) and fungivorous nematodes increased but parasitism of nematodes by H. rhossiliensis usually decreased with addition of wheat straw or composted cow manure to a loamy sand naturally infested with H. rhossiliensis. These results do not support the hypothesis that organic amendments will enhance parasitism of nematodes by H. rhossiliensis.     

Repeated Sampling to Determine the Precision of Estimating Nematode Population Densities

The first phase of this study involved repeated samplings of five fields using composite samples of 10, 20, 40, and 80 soil cores, to determine the precision of nematode assays. The second phase focused on randomly selecting two and four 2-ha subunits (data on Meloidogyne spp.) of 24 fields ranging from 6 to 40 ha and computing the precision of estimated means for these numbers ofsubunits versus the general field mean (based on all 2-ha subunits). Average numbers of nematodes from most samples containing Meloidogyne spp., Heterodera glycines, Helicotylenchus dihystera, Scutellonema brachyurum, and (or) Hoplolaimus galeatus were within 50% of the overall means. Coefficient of variation (CV) values were generally lower for 40 cores than for 10, 20, and 80 cores per sample. When data for all nematodes and fields were combined, this value was lowest for 40 and 80 cores. The CV values were higher for Meloidogyne spp. than for H. glycines. Means of two samplings increased the probability of obtaining numbers nearer the mean for that field than numbers from a single composite sample. For the second phase, population estimates of Meloidogyne spp. based on four 2-ha subunits generally were closer to field means than were those for two subunits. Sampling precision with these subunits diminished greatly in large fields with variable soils and (or) mixed cropping histories. Either two or four subunits gave population estimates within 3-20% of the field mean in most instances. The mean man hours required for sampling ca. 2-ha parcels of 4-20-ha fields was 0.54 hours.     

Preliminary laboratory tests with two species of entomophilic nematodes for control of Musca domestica in intensive animal units‡

Two species of entomophilic nematodes, Heterorhabditis heliothidis (NZ strain) and Steinernema feltiae (Agriotos strain) were tested in the laboratory against immature and adult stages of a strain (G) of Musca domestica with multiple insecticide-resistance. Both species of nematodes killed larval stages of M. domestica on inoculated filter papers. S. feltiae was the most virulent parasite, and killed >90% of all larval stages at the two highest doses of 25 000 and 50 000 nematodes per 0·5 ml tap water. No puparia were parasitised by either species. There was no parasitism of the larval stages after exposure to chicken manure treated with nematodes. All adult flies were parasitised after they were exposed to bait-pads previously inoculated with S. feltiae, 93.3% were parasitised by H. heliothidis.     

Spatial Patterns of Entomopathogenic Nematodes in Microcosms: Implications for Laboratory Experiments

Laboratory microcosms were used to: i) measure the effects of soil moisture on survival of Steinernema riobravis and ii) investigate the suitability of using microcosms to study motility and survival of these nematodes. Nematodes recovered from soil contained in petri dishes declined by more than 95% during 7 days, whereas nematodes recovered from the inner surfaces of dishes increased 35-fold. After 7 days in dishes, >20 times as many nematodes were recovered from dish surfaces than from soil. Nematodes exhibited a negative geotropism; greater numbers of nematodes were recovered from the lid surfaces than from the surfaces of dishes. Survivorship of nematodes in soil in plastic centrifuge tubes was somewhat greater than in petri dishes, and fewer nematodes ascended above the soil line in tubes than dishes. Downward migration of nematodes was inversely related to soil column diameter, possibly due to relatively unimpeded movement along container surfaces. An assay was developed by which nematodes were rinsed from the inner surfaces of centrifuge tubes into the soil. The resulting slurry was then processed on Baermann trays to recover motile nematodes. Nematode survival in soil in centrifuge tubes was higher at soil moistures between 2-4% than at lower (0.5-1.0%) and higher (4.0-12.0%) moisture levels. Survival of S. riobravis may be enhanced by quiescence induced by moisture deficits.     

Susceptibility of the Colorado Potato Beetle and the Sugarbeet Wireworm to Steinernema feltiae and S. glaseri

In laboratory tests, larvae of the Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say), and the sugarbeet wireworm (SBW), Limonius californicus (Mannerheim), were exposed to the nematodes Steinernema feltiae Filipjev (Mexican strain) (= Neoaplectana carpocapsae) and S. glaseri Steiner in soil. S. feltiae caused significantly higher mortality in SBW larvae than did S. glaseri, but both nematode species were equally effective against CPB larvae. The minimum concentration of S. feltiae for 100% mortality of CPB larvae after 13 days was 157 nematodes/cm² of soil, and the LC₅₀ based on 6-day mortality was 47.5 nematodes/cm²; in contrast, 100% mortality of SBW larvae was not achieved with even the highest concentration tested, 393 nematodes/cm². CPB adults emerging from nematode-contaminated soil were not infected. In field cage tests, S. feltiae applied to the soil surface at the rates of 155 and 310 nematodes/cm² soil caused 59% and 71% mortality, respectively, of late-fourth-instar spring-generation CPB, and 28% and 29% mortality, respectively, of SBW. No infection was obtained when larvae of summer generation CPB and SBW were placed in the same cages approximately 6 weeks after nematodes were applied to the soil. Inundative soil applications of S. feltiae, though cost prohibitive at present, were effective in reducing caged CPB and SBW field populations.     

Suppression of Root-knot Nematode Populations with Selected Rapeseed Cultivars as Green Manure

Meloidogyne chitwoodi races 1 and 2 and M. hapla reproduced on 12 cultivars of Brassica napus and two cultivars of B. campestris. The mean reproductive factors (Rf), Rf = Pf at 55 days ÷ 5,000, for the three nematodes were 8.3, 2.2, and 14.3, respectively. All three nematodes reproduced more efficiently (P < 0.05) on B. campestris than on B. napus. Amending M. chitwoodi-infested soil in plastic bags with chopped shoots of Jupiter rapeseed reduced the nematode population more (P < 0.05) than amendment with wheat shoots. Incorporating Jupiter shoots to soil heavily infested with M. chitwoodi in microplots reduced the nematode population more (P < 0.05) than fallow or corn shoot treatments. The greatest reduction in nematode population density was attained by cropping rapeseed for 2 months and incorporating it into the soil as a green manure.     

Effects of Insecticides on Movement,Nictation, and Infectivity of Steinernema carpocapsae

Movement, nictation, and infectivity of Steinernema carpocapsae strain All were compared for ensheathed (EnJ) and desheathed (DeJ) infective juveniles exposed to the insecticides acephate, dichlorvos, methomyl, oxamyl, or permethrin. Nematode response to various solutions included normal sinusoidal movement, uncoordinated motion, twitching, convulsion or formation of a pretzel shape, an inactive "S" posture with fine twitching, or a quiescent straight posture. The DeJ displayed these movements at lower concentrations of each insecticide than did EnJ. In petri dish bioassays, insecticide-treated EnJ caused generally lower mortality in the common cutworm, Spodoptera litura, than did EnJ alone but caused greater insect mortality than did insecticides alone. Nematode response to chemicals was more clearly demonstrated by nictating behavior than by the movement bioassay. Nictation of DeJ was suppressed by the test chemicals at low concentrations, except for acephate and permethrin. Nictating EnJ or DeJ, regardless of chemical treatment, killed host insects faster than did non-nictating juveniles. Insecticides that enhance nictating behavior at certain concentrations may be used for mixed applications with nematodes.     

Ingestion,Retention, and Transmission of two Strains of Raspberry Ring-spot Virus by Longidorus macrosoma

The transmission of two strains of raspberry ringspot virus (RRV) by small numbers of nematodes was compared. A strain of RRV from Scotland (RRV-S), originally found in the field associated with Longidorus elongatus, was transmitted frequently by L. elongatus but only once by L. macrosoma. A strain from England (RRV-E) associated with L. macrosoma in the field was transmitted infrequently by each species of nematode. The reasons why L. macrosoma infected only a small proportion of bait plants with virus were investigated, and it was found that most of the nematodes tested had fed on the source plants and many had ingested virus. Most nematodes exposed to RRV-E or RRV-S had fed on the roots of the bait plants and, when thin sections were examined by electron microscope, had retained particles (thought to be those of the virus) in the region of the anterior odontostyle, Thus, most nematodes seem to have had ample opportunity to transmit virus, and the low frequency of transmission may have been due to a failure of the virus particles to be released from the site of retention or to a lack of infectivity of the virus when L. macrosoma was the vector and Petunia hybrida was the host.     

Field Evaluation of Steinernema feltiae Against the Web-spinning Larch Sawfly Cephalcia lariciphila

Field trials were conducted in Rheola Forest, Wales, Great Britain, to determine the effectiveness of Steinernema feltiae UK strain in controlling the web-spinning larch sawfly Cephalcia lariciphila. Foliar sprays at the rate of 5,000-20,000 nematodes/100 cm branch resulted in 3.4-29.4% infection of sawfly larvae. Soil application of 200 nematodes/cm² resulted in 61% infection of sawfly prepupae and 17.3% of pupae. Prepupal infection ranged from 4.8 to 14.7% 1 year after nematode application. Soil applications of this nematode show that it has potential for biological control of sawfly prepupae.     

Use of a Stilbene Brightener,Tinopal LPW,as a Radiation Protectant for Steinernema carpocapsae

A stilbene fluorescent brightener, Tinopal LPW, was used as an ultraviolet (UV) protectant for the entomopathogenic nematode Steinernema carpocapsae (All strain). Irradiation of an aqueous suspension of nematodes produced a LC₅₀ in 15.7 minutes under a sunlamp and in 31.7 minutes in direct sunlight. Irradiation by both sunlamp and sunlight of a suspension of nematodes in Tinopal LPW did not reduce their biological activity as measured by their ability to parasitize wax moth larvae after exposure of 8 hours and 4 hours, respectively. Tinopal LPW appeared promising as a radiation protectant.     

Sterol Composition and Ecdysteroid Content of Eggs of the Root-knot Nematodes Meloidogyne incognita and M. arenaria

Free and esterified sterols of eggs of the root-knot nematodes Meloidogyne incognita races 2 and 3 and M. arenaria race 1 were isolated and identified by gas-liquid chromatography-mass spectrometry. The major sterols of eggs of each race were 24-ethylcholesterol (33.4-38.8% of total sterol), 24-ethylcholestanol (18.3-25.3%), 24-methylcholesterol (8.6-11.7%), 24-methylcholestanol (7.7-12.5%), and cholesterol (4.6-11.6%). Consequently, the major metabolic transformation performed by Meloidogyne females or eggs upon host sterols appeared to be saturation of the sterol nucleus. The free and esterified sterols of the same race did not differ appreciably, except for a slight enrichment of the steryl esters in cholesterol. Although the sterol composition of Meloidogyne eggs differed from that of other life stages of other genera of plant-parasitic nematodes, the three Meloidogyne races could not be distinguished from each other by their egg sterols. Ecdysteroids, compounds with hormonal function in insects, were not detected by radioimmunoassay in the Meloidogyne eggs either as free ecdysteroids or as polar conjugates.     

Comparison of Two Steinernematid Species for Control of the Root Weevil Diaprepes abbreviatus

Steinernema carpocapsae Weiser All strain was compared to Steinernema riobravis Cabanillas, Poinar, and Raulston for control of the root weevil, Diaprepes abbreviatus (L.), in the laboratory and in potted citrus. In the laboratory bioassay, D. abbreviatus larvae were exposed to 30, 60, and 120 nematodes/cm³ in sand. Insect mortality 1 week after application was greater (P ≤ 0.05) for S. riobravis than for S. carpocapsae in the laboratory bioassay. In the greenhouse bioassay, D. abbreviatus larvae were exposed to 3 and 9 nematodes per cm³ of soil in potted citrus. Again, at each rate, mortality was greater (P ≤ 0.05) in pots treated with S. riobravis than in pots treated with S. carpocapsae. The results of this study suggest that S. riobravis is a better biological control agent against D. abbreviatus larvae in potted plants than S. carpocapsae.     

An Assay for Hirsutella rhossiliensis Spores and the Importance of Phialides for Nematode Inoculation

A spore assay was developed to measure the relative density of spores of the nematophagous fungus Hirsutella rhossiliensis in soil. Orchard soil containing H. rhossiliensis-parasitized Criconemella xenoplax was placed in vials and incubated for 0-120 days before the addition of probe nematodes, Heterorhabditis heliothidis juveniles. After 18 hours, H. heliothidis were extracted from the soil and examined for adhering spores of H. rhossiliensis. No spores were detected when H. heliothidis were added to freshly mixed soil, but the percentage of H. heliothidis with spores increased rapidly if soil was incubated undisturbed. Because mixing soil detaches spores from phialides, the results indicate that spores must be attached to phialides to adhere to nematodes. The spore assay was compared with a plate assay that measures the population density of H. rhossiliensis-parasitized C. xenoplax. Results from the two assays were highly correlated, suggesting that spores occur in three phases: reserves in nematodes that may be converted into spores; spores on phialides and therefore capable of adhering to nematodes; and spores detached from phialides and thus incapable of adhering to nematodes.     

Pathogenicity of Bursaphelenchus xylophilus on Pines in Minnesota and Wisconsin

The pinewood nematode, Bursaphelenchus xylophilus, was inoculated into established native jack and red pines (Pinus banksiana and P. resinosa) and exotic Austrian pine (P. nigra) in Minnesota and Wisconsin forests during summer 1981. The nematode isolates did not kill established nonstressed pine trees growing in the forest. However, the same nematode isolates killed pine seedlings under greenhouse conditions. Girdling the main stem of some trees to induce stress resulted in the death of the majority of inoculated and noninoculated branches of Austrian and jack pines, but no branch death was observed on red pine. Greater numbers of nematodes were extracted from branches of inoculated, girdled trees than from nongirdled trees. The mean number of nematodes extracted from branches of inoculated, nongirdled trees was 0.3 - 14 nematodes per gram of wood.     

Characterization of a <Emphasis Type="Italic">Pseudomonas fluorescens</Emphasis> strain from tomato rhizosphere and its use for integrated management of tomato damping-off

A highly antagonistic Pseudomonas fluorescens strain was isolated from tomato rhizosphere and characterized for its in vitro and in vivo biocontrol potential against Pythium aphanidermatum. The identified Pseudomonas fluorescens strain (PfT-8) was capable of producing high levels of chitinase, β-1,3-glucanase, cellulase, fungitoxic metabolites and siderophores. Seven different carrier formulations including a talc-based powder, lignite-based powder, peat-based powder, lignite + fly ash-based powder, wettable powder, bentonite-paste and polyethylene glycol (PEG) paste were prepared utilizing PfT-8. Shelf life was evaluated for up to 6 months of storage at ambient room temperature (28 °C). Biocontrol efficacy of formulations was studied under greenhouse and field conditions. The formulations were stable up to at least 2 months of storage at ambient room temperature. Among the formulations, peat, lignite, lignite+fly-ash and bentonite paste based formulations maintained higher propagule number than others and also showed greater biocontrol potential. However, propagule number gradually decreased with time. Several organic amendments including farm yard manure (FYM), leaf-compost, poultry manure, press mud, vermi-compost and neem cake were incorporated into soil to study their influence on P. fluorescens colonization in the rhizosphere and on potential disease control. Soil incorporation of organic amendments and specifically poultry manure and FYM, significantly reduced damping-off incidence and also augmented the rhizosphere population of the marked␣P.␣fluorescens strain that was resistant to streptomycin and rifampicin. An integrated␣approach of damping-off management employing dual inoculation of PfT-8 in seed and soil coupled with soil application of organic amendments including poultry manure or␣FYM was evaluated under field conditions. Under these conditions, damping-off incidence substantially reduced by up to 90% and further the healthy plant stand, plant biomass and plant rhizosphere population of P. fluorescens increased significantly.     

Development and Fecundity of Reesimermis nielseni,a Nematode Parasite of Mosquitoes

Maturation of the mermithid nematode Reesimermis nielseni to the adult stage began by the tenth day after emergence of the nematodes from their hosts at ambient temperatures (24-27 C). Most postparasitic males and females reached the adult stage after 50 and 70 days, respectively. The first females exhibiting egg development and oviposition were observed 25-30 days after emergence, but some oviposition was still taking place 150 days later. Reesimermis nielseni laid an average 2,480 eggs per female over an 18-day oviposition period. A majority of the mature eggs hatched within 7 h after the cultures were flooded. The preparasites are short-lived, and only a few were able to infect exposed hosts after 72 h.