Entomopathogenic Nematodes Are Not an Alternative to Fenamiphos for Management of Plant-Parasitic Nematodes on Golf Courses in Florida

With the cancellation of fenamiphos in the near future, alternative nematode management tactics for plant-parasitic nematodes (PPN) on golf courses need to be identified. The use of entomopathogenic nematodes (EPN) has been suggested as one possible alternative. This paper presents the results of 10 experiments evaluating the efficacy of EPN at managing PPN on turfgrasses and improving turf performance. These experiments were conducted at various locations throughout Florida over the course of a decade. In different experiments, different EPN species were tested against different species of PPN. Separate experiments evaluated multiple rates and applications of EPN, compared different EPN species, and compared single EPN species against multiple species of PPN. In a few trials, EPN were associated with reductions in certain plant-parasite species, but in other trials were associated with increases. In most trials, EPN had no effect on plant parasites. Because EPN were so inconsistent in their results, we conclude that EPN are not acceptable alternatives to fenamiphos by most turf managers in Florida at this time.     

Vertical Distribution of Plant-parasitic Nematodes in Sandy Soil under Soybean

Vertical distribution of five plant-parasitic nematodes was examined in two north Florida soybean fields in 1987 and 1988. Soil samples were collected from 0-15 cm, 15-30 cm, and 30-45 cm deep at each site. Soil at the three depths consisted of approximately 96% sand. More than 50% of Belonolaimus longicaudatus population densities occurred in the upper 15-cm soil layer at planting, but the species became more evenly distributed through the other depths as the season progressed. Criconemella sphaerocephala was evenly distributed among the three depths in one field but was low (< 20% of the total density) in the upper 15 cm at a second site. Maximum population densities of Pratylenchus brachyurus were observed at 15-30 cm on most sampling dates. Vertical distributions of Meloidogyne incognita and Paratrichodorus minor were erratic and showed seasonal variation. A diagnostic sample from the upper 0-15 cm of these soybean fields revealed only a minority of the populations of most of the phytoparasitic species present.     

Temporary Changes in Populations of Soil Organisms after Field Application of Entomopathogenic Nematodes

To assess the effect of an inundative release of entomopathogenic nematodes on soil organisms, population densities of soil-dwelling organisms were monitored before and after an application of an aqueous suspension of Heterorhabditis megidis to field plots in mown grassland (Exp. I) at a level of 0.38 million/m² and to plots (Exp. II) situated in a forested area, a grass sports field and an orchard at a level of 1.5 million/m². At the forested site, heat-killed H. megidis (1.5 million/m²) also were applied to two plots to compare the impact on soil organisms of a large introduction of living and dead nematodes. Post-treatment, temporary changes in natural population densities of several nematode genera and other organisms were detected in H. megidis-treated plots in both experiments. Temporary changes in the nematode trophic structure occurred in the percentages of nematode omnivores, herbivores and predators in both experiments. Evidence from all sites suggests that the changes were temporary and that the presence of decaying H. megidis following treatment contributed to nutrient enrichment of the soil and to direct and indirect effects on the nematode community.     

Effects of Nematophagous Fungi on Numbers and Death Rates of Bacterivorous Nematodes in Arable Soil

In a series of microcosm experiments with an arable, sandy loam soil amended with sugarbeet leaf, the short-term (8 weeks) dynamics of numbers of nematodes were measured in untreated soil and in γ-irradiated soil inoculated with either a field population of soil microorganisms and nematodes or a mixed population of laboratory-propagated bacterivorous nematode species. Sugarbeet leaf stimulated an increase in bacterivorous Rhabditidae, Cephalobidae, and a lab-cultivated Panagrolaimus sp. Differences were observed between the growth rates of the nematode population in untreated and γ-irradiated soils, which were caused by two nematophagous fungi, Arthrobotrys oligospora and Dactylaria sp. These fungi lowered the increase in nematode numbers due to the organic enrichment in the untreated soil. We estimated the annually produced bacterivous nematodes to consume 50 kg carbon and 10 kg nitrogen per ha, per year, in the upper, plowed 25 cm of arable soil.     

Distribution of Field Corn Roots and Parasitic Nematodes in Subsoiled and Nonsubsoiled Soil

A field trial was conducted for 2 years in an Arredondo fine sand containing a tillage pan at 15-20 cm deep to determine the influence of subsoiling on the distribution of corn roots and plant-parasitic nematodes. Soil samples were taken at various depths and row positions at 30, 60, and 90 days after planting in field corn subsoiled under the row with two chisels and in non-subsoiled corn. At 30 and 60 days, in-row nematode population densities to 60 cm deep were not affected by subsoiling compared with population densities in nonsubsoiled plots. After 90 days, subsoiling had not affected total root length or root weight at the 20 depth-row position sampling combinations, but population densities of Meloidogyne incognita and Criconemella spp. had increased in subsoiled corn. Numbers of Pratylenchus zeae were not affected. Subsoiling generally resulted in a change in distribution of corn roots and nematodes in the soil profile but caused little total increase in either roots or numbers of nematodes. Corn yield was increased by subsoiling.     

From Augmentation to Conservation of Entomopathogenic Nematodes: Trophic Cascades, Habitat Manipulation and Enhanced Biological Control of Diaprepes abbreviatus Root Weevils in Florida Citrus Groves

The use of entomopathogenic nematodes (EPN) for management of the root weevil, Diaprepes abbreviatus, in Florida citrus groves is considered a biological control success story and typically involves augmentation in which EPN are applied inundatively as biopesticides to quickly kill the pest. However, recent evidence indicates that efficacy of EPN applications in Florida citrus depends on soil type. They are very effective in the well drained coarse sands of the Central Ridge but often less so in poorly drained fine-textured soils of the Flatwoods. Moreover, groves on the Central Ridge can harbor rich communities of endemic EPN that might often suppress weevil populations below economic thresholds, whereas Flatwoods groves tend to have few endemic EPN and frequent weevil problems. Current research is examining the ecological dynamics of EPN in Florida citrus groves, the potential impact of EPN augmentation on soil food webs, especially endemic EPN, and whether habitat manipulation and inoculation strategies might be effective for conserving and enhancing EPN communities to achieve long-term control in problem areas. Conservation biological control could extend the usefulness of EPN in Florida citrus and be especially appropriate for groves with persistent weevil problems.     

Effects of the Integration of Sunn Hemp and Soil Solarization on Plant-Parasitic and Free-Living Nematodes

Sunn hemp (SH), Crotolaria juncea, is known to suppress Rotylenchulus reniformis and weeds while enhancing free-living nematodes involved in nutrient cycling. Field trials were conducted in 2009 (Trial I) and 2010 (Trial II) to examine if SH cover cropping could suppress R. reniformis and weeds while enhancing free-living nematodes if integrated with soil solarization (SOL). Cover cropping of SH, soil solarization, and SH followed by SOL (SHSOL) were compared to weedy fallow control (C). Rotylenchulus reniformis population was suppressed by SHSOL at the end of cover cropping or solarization period (Pi) in Trial I, but not in Trial II. However, SOL and SHSOL did not suppress R. reniformis compared to SH in either trial. SH enhanced abundance of bacterivores and suppressed the % herbivores only at Pi in Trial II. At termination of the experiment, SH resulted in a higher enrichment index indicating greater soil nutrient availability, and a higher structure index indicating a less disturbed nematode community compared to C. SOL suppressed bacterivores and fungivores only in Trial II but not in Trial I. On the other hand, SHSOL enhanced bacterivores and fungivores only at Pi in Trial I. Weeds were suppressed by SH, SOL and SHSOL throughout the experiment. SHSOL suppressed R. reniformis and enhanced free-living nematodes better than SOL, and suppressed weeds better than SH.     

Anhydrobiotic Coiling of Nematodes in Soil

Nematodes of three genera (Acrobeloides sp., Aphelenchus avenae, and Scutellonema brachyurum) were induced to coil and enter anhydrobiosis in drying soil of two types: sandy loam and loamy sand. Coiling was studied in relationship to soil moisture characteristics. Coiling and the physiological state of anhydrobiosis occurred before the water in sandy soils reached a water potential of -15 bars. Coiling was maximum at 3-6 bars, depending on the soil type and nematode species. It appeared that induction of coiling and anhydrohiosis were determined by the physical forces exerted by the water film surrounding the nematode, which, for these three species, was 6-9 monomolecular layers of water, rather than the % moisture and relative humidity of the soil per se.     

Comparison of Three Methods for Estimating the Number of Entomopathogenic Nematodes Present in Soil Samples

Numbers of Steinernema sp. (CB2B) and S. carpocapsae (Agriotos) exponentially declined after application into a clay loam soil. Over a 35-day sampling period, Steinernema sp. (CB2B) was more persistent than S. carpocapsae (Agriotos). The presence or absence of the second-stage cuticle on the third-stage juveniles (J3) at the time of application did not alter the rate of population decline of Steinernema sp. (CB2B). Nearly all J3 of Steinernema sp. (CB2B) and S. carpocapsae (Agriotos) lost their cuticle within 24 hours of being in soil. Centrifugal flotation recovered the greatest number of nematodes, with a lower variance than either the live bait or Baermann funnel techniques. A strong positive linear relationship was evident between numbers of nematodes present in the soil and the numbers that established in a bait insect. Approximately 40% of Steinernema sp. (CB2B) and 30% of the S. carpocapsae (Agriotos) present in the soil established in Galleria mellonella larvae. The extraction techniques had different efficiencies and gave different relative estimates of persistence for the two species. Persistence and infectivity was best measured using a combination of live bait and flotation techniques.     

Molecular Analysis of the Interactions between Cyst Nematodes and Their Hosts

In order to complete its life cycle, a cyst nematode must stimulate the production of a specialized syncytial feeding site within host root tissues. This process is characterized by major changes in local root morphology, including enlargement of affected nuclei and nucleoli, cell wall degradation, and proliferation of subcellular organelles. At the molecular level very little is known about the processes involved in this host response, but recent evidence suggests that cyst nematodes are able to regulate specific host genes. The host-parasite model system provided by Arabidopsis thaliana and Heterodera schachtii will be fundamental to our future understanding of the formation of syncytia. Molecular biology now offers us the opportunity to study this complex host-parasite interaction in great detail. A better understanding of the host genes regulated by cyst nematodes and the mechanisms by which this regulation is achieved will facilitate the engineering of crop cultivars that possess novel forms of resistance to these adept parasites.     

Quarantine host range studies with Lophyrotoma zonalis, an Australian sawfly of interest for biological control of melaleuca, Melaleuca quinquenervia, in Florida, U.S.A.

The Australian melaleuca tree, Melaleuca quinquenervia (Cav.) S. T. Blake (Myrtaceae), has naturalized in southern Florida,U.S.A., and is now one of that regions most important weeds.Primarily a weed of wetlands, it also infests neighboring drierareas. Current efforts to restore the South Florida ecosystem arethreatened by the continuing range expansion of melaleuca andother weeds. In an effort to supplement the current chemical andcultural control methods for melaleuca, a search for potentialbiological control agents was begun in Australia in 1986. Thesawfly, Lophyrotoma zonalis, was determined after extensive fieldand laboratory studies to have potential as a biological controlagent. Larvae of L. zonalis eat leaves and occasionally defoliatelarge trees in Australia. Host range studies were conducted in aFlorida quarantine facility with native and cultivated plantspecies. Multi-choice and no-choice oviposition tests wereconducted with 36 species in the Myrtaceae and with 18 species inother families. Larvae developed to prepupae and adults from theeggs oviposited on 23 species of Myrtaceae only on 3 species ofbottlebrushes, Callistemon. Medium-sized larvae were tested forfeeding on bouquets of plant cuttings and on potted plants. Theyare the stage that might wander from defoliated trees. Noticeablefeeding, but much less than on melaleuca, was restricted to theMyrtaceae, except for a few individual larvae that fed on waxmyrtle, Myrica cerifera. Medium-sized larvae became prepupae onlyon Melaleuca decora (73%) and on wax myrtle (10%). However,neither species received eggs in the oviposition tests. Thesestudies confirmed the narrow host range of L. zonalis aspreviously reported from field and laboratory studies inAustralia.     

Potential for successful population establishment of the nonindigenous sacred ibis in the Florida Everglades

The nonindigenous sacred ibis (Threskiornis aethiopicus) was first discovered breeding in the Florida Everglades in 2005 in the Arthur R. Marshall Loxahatchee National Wildlife Refuge. Prior to this, sacred ibises were seen periodically throughout South Florida since the mid 1990s, with occasional confirmed breeding occurrences in Miami-Dade and Palm Beach counties. We used a logistic regression model developed by Allen (Biol Invasions 8:491–500, 2006) to predict the probability of successful establishment of sacred ibis in the Florida Everglades ecosystem. Empirical data collected from several sacred ibis nests and chicks were used to validate those findings. The probability of successful establishment was estimated to be 73%. The physiological condition of nestlings suggested that this species was able to fledge chicks in good condition, thus adding to the potential to increase their breeding population. Exponential population growth rates and expanding distribution of the nonindigenous sacred ibis in France demonstrate this species’ potential for becoming invasive in Florida. We suggest that the most prudent and effective management strategy is eradication of the few pioneering individuals that are nesting in the Everglades as well as the urban source population.     

Strip-tilled Cover Cropping for Managing Nematodes, Soil Mesoarthropods, and Weeds in a Bitter Melon Agroecosystem

A field trial was conducted to examine whether strip-tilled cover cropping followed by living mulch practice could suppress root-knot nematode (Meloidogyne incognita) and enhance beneficial nematodes and other soil mesofauna, while suppressing weeds throughout two vegetable cropping seasons. Sunn hemp (SH), Crotalaria juncea, and French marigold (MG), Tagetes patula, were grown for three months, strip-tilled, and bitter melon (Momordica charantia) seedlings were transplanted into the tilled strips; the experiment was conducted twice (Season I and II). Strip-tilled cover cropping with SH prolonged M. incognita suppression in Season I but not in Season II where suppression was counteracted with enhanced crop growth. Sunn hemp also consistently enhanced bacterivorous and fungivorous nematode population densities prior to cash crop planting, prolonged enhancement of the Enrichment Index towards the end of both cash crop cycles, and increased numbers of soil mesoarthropods. Strip-tilled cover cropping of SH followed by clipping of the living mulch as surface mulch also reduced broadleaf weed populations up to 3 to 4 weeks after cash crop planting. However, SH failed to reduce soil disturbance as indicated by the Structure Index. Marigold suppressed M. incognita efficiently when planted immediately following a M. incognita-susceptible crop, but did not enhance beneficial soil mesofauna including free-living nematodes and soil mesoarthropods. Strip-tilled cover cropping of MG reduced broadleaf weed populations prior to cash crop planting in Season II, but this weed suppression did not last beyond the initial cash crop cycle.     

Effect of Compost and Manure Soil Amendments on Nematodes and on Yields of Potato and Barley: A 7-Year Study

A 7-year study located in Prince Edward Island, Canada, examined the influence of compost and manure on crop yield and nematode populations. The compost used in this study consisted of cull waste potatoes, sawdust, and beef manure in a 3:3:1 ratio, respectively. No plant-parasitic nematodes were detected in samples collected from windrow compost piles at 5- and 30-cm depths prior to application on field plots. Low population densities of bacterial-feeding nematodes were recovered from compost windrows at the 5-cm depth. Field plots of potato (Solanum tuberosum cv. Kennebec) received compost applied at 16 metric tonnes per hectare, or beef manure applied at 12 metric tonnes per hectare. An adjacent trial with barley (Hordeum vulgare cv. Mic Mac) received only the compost treatment. In both trials the experimental design was a complete randomized block with four replicates. Data averaged over seven growing seasons indicated that population levels of root-lesion nematodes (primarily Pratylenchus penetrans) were higher in root-zone soil in potato plots treated with either compost or manure compared to the untreated control plots. The soil amendments did not affect root-knot nematode (Meloidogyne hapla) population densities in the potato plots, but clover-cyst nematodes (Heterodera trifolii) were more numerous in the root-zone soils of barley treated with compost compared to the untreated plots. Numbers of bacterial-feeding nematodes (primarily Diplogaster lheritieri) were greater in soil in potato plots treated with manure and in soil around barley roots than in untreated plots. Total yields of potato tubers averaged over seven growing seasons increased by 27% in the plots treated with either compost or manure. Grain yields of barley also were increased by 12% when compost was applied. These results indicated that organic amendments increased crop yields, but the impacts on different nematode species varied and usually increased soil population levels.     

Estimating Sample Size and Persistence of Entomogenous Nematodes in Sandy Soils and Their Efficacy Against the Larvae of Diaprepes abbreviatus in Florida

In two studies to estimate sampling requirements for entomogenous nematodes in the field, highest persistence of Heterorhabditis bacteriophora after application occurred beneath the canopies of mature citrus trees. Nematode persistence declined with distance from the center-line of the tree row toward the row-middles. Immediately after nematode application to soil, 32 samples (15 cm deep, 2.5-cm diameter) beneath a single tree were required to derive 95% confidence intervals that were within 40% of mean nematode population density. The estimated probability of measuring the mean density within 40%, using 32 samples, declined to 88% at 2 days post-application and to 76% at 7 days. The persistence in soil of Steinernema carpocapsae, S. riobravis, and two formulations containing H. bacteriophora and their efficacy against the larvae of Diaprepes abbreviatus were compared in a grove of 4-year-old citrus trees. Within 6 days, the recovered population densities of all nematodes declined to <5% of levels on day 0. The recovery of H. bacteriophora during the first 2 weeks was lower than that of the other two species. Steinemema riobravis and both formulations of H. bacteriophora reduced recovery of D. abbreviatus by more than 90% and 50%, respectively. Steinernema carpocapsae did not affect population levels of the insect.     

Accelerated Movement of Nematodes from Soil in Baermann Funnels with Temperature Gradients

Baermann funnels were modified to eliminate or reverse the small temperature gradient (1-2 C/cm) across the soil layer that normally results from water evaporation. Effects of modifications on extraction efficiency were examined at various ambient temperatures and after overnight adaptation of three nematode species at 20 and 30 C. Extraction of Meloidogyne incognita from sandy loam, Tylenchulus semipenetrans from sandy clay loam, and Rotylenchulus reniformis from silt was greatly accelerated simply by covering funnels to prevent evaporation. In most cases, covering increased the nematodes extracted by 10-100 times after 5.5-48 hours. Faster and more efficient extraction of R. reniformis occurred over a wide range of ambient temperature (18-29 C). Effects of ambient temperature and temperature gradient direction on Baermann funnel extraction of R. reniformis were partly inconsistent with the behavior of R. reniformis in agar. Nematodes in agar moved toward cold at some ambient temperatures and toward heat at other temperatures. They always appeared to move toward cold on Baermann funnels. Differences were not attributable to blockage of gas exchange by covers. In agar and in funnels, the patterns of response to ambient temperature were shifted in the direction of the storage temperature.     

Impact of a Nematode-parasitic Fungus on the Effectiveness of Entomopathogenic Nematodes

The impact of the nematode-parasitic fungus Hirsutella rhossiliensis on the effectiveness of Steinernema carpocapsae, S. glaseri, and Heterorhabditis bacteriophora against Galleria mellonella larvae was assessed in the laboratory. The presence of Hirsutella conidia on the third-stage (J3) cuticle of S. carpocapsae and H. bacteriophora interfered with infection of insect larvae. Conidia on the J3 cuticle of S. glaseri and on the ensheathing second-stage cuticle of H. bacteriophora did not reduce the nematodes'' ability to infect larvae. The LD₅₀ values for S. carpocapsae, S. glaseri, and H. bacteriophora in sand containing H. rhossiliensis were not different from those in sterilized sand when Galleria larvae were added at the same time as the nematodes. However, when Galleria larvae were added 3 days after the nematodes, the LD₅₀ of S. glaseri was higher in Hirsutella-infested sand than in sterilized sand, whereas the LD₅₀ of H. bacteriophora was the same in infested and sterilized sand. Although the LD₅₀ of S. carpocapsae was much higher in Hirsutella-infested sand than in sterilized sand, the data were too variable to detect a significant difference. These data suggest that H. bacteriophora may be more effective than Steinernema species at reducing insect pests in habitats with abundant nematode-parasitic fungi.     

Influence of Water and Soil Temperature on the Concentration and Efficacy of Phenamiphos for Control of Root-Knot Nematodes

Field plots of Tifton loamy sand were treated with phenamiphos for control of root-knot nematodes in a multiple-crop system of turnips, field corn, and southern peas. Preplant applications of phenamiphos protected roots of turnips and corn from damage by root-knot nematodes. Concentrations of phenamiphos at application in the 0-15-cm soil layer were near 6 μg/g on turnips and near 4 μg/g on corn and southern peas. After 30 d, concentrations were approximately 1 μg phenamiphos/g of soil for all crops. Concentrations of 2.0-3.8 μg phenamiphos/g of soil for 9-d duration appeared to be adequate for control of root-knot nematodes on field corn and southern peas in this multiple-crop system. Stepwise regression analyses indicated that 31%, 62%, and 22% of the variations in concentration of phenamiphos in the soil planted to turnips, corn, and southern peas, respectively, were attributable to the amount of water that the plots received. Soil temperature had no effect on concentrations of phenamiphos.     

Impact of Soil Texture on the Reproductive and Damage Potentials of Rotylenchulus reniformis and Meloidogyne incognita on Cotton

The effects of soil type and initial inoculum density (Pi) on the reproductive and damage potentials of Meloidogyne incognita and Rotylenchulus reniformis on cotton were evaluated in microplot experiments from 1991 to 1993. The equilibrium nematode population density for R. reniformis on cotton was much greater than that of M. incognita, indicating that cotton is a better host for R. reniformis than M. incognita. Reproduction of M. incognita was greater in coarse-textured soils than in fine-textured soils, whereas R. reniformis reproduction was greatest in a Portsmouth loamy sand with intermediate percentages of clay plus silt. Population densities of M. incognita were inversely related to the percentage of silt and clay, but R. reniformis was favored by moderate levels of clay plus silt (ca. 28%). Both M. incognita races 3 and 4 and R. reniformis effected suppression of seed-cotton yield in all soil types evaluated. Cotton-yield suppression was greatest in response to R. reniformis at high Pi. Cotton maturity, measured as percentage of open bolls at different dates, was affected by the presence of nematodes in all 3 years.