Susceptibility of Sap Beetles (Coleoptera: Nitidulidae) to Entomopathogenic Nematodes

Nitidulid beetles (Coleoptera) are considered to be serious pests of date palms throughout the world. They attack ripe fruit, causing it to rot, and damage is reflected in both reduced yield and lower fruit quality. The present study was aimed at an evaluation of the susceptibility to different sap beetles to entomopathogenic nematodes. We further tested nematode efficacy in pots filled with soil infested by third instar larvae of the two beetle species. In Petri dish assay, mortality levels of Carpophilus humeralis and C. hemipterus exposed to Heterorhabditis sp. IS-5 strain indicated that the latter is less susceptible to nematode infection. Exposure of both sap beetle species to different nematode strains gave moderate levels of mortality (35-65%) with the heterorhabditid strains HP88, IS-5 and IS-25. The IS-12 strain of Heterorhabditis sp. showed poor virulence (<35% mortality) against larvae of C. humeralis as well as larvae and pupae of C. humipterus. The nematode species S. riobrave showed moderate virulence (35-65%) mortality to larvae and pupae of S. humeralis as well as to larvae of C. hemipterus . Exposure of C. hemeralis to different concentrations of Heterorhabditis sp. IS-5 in pots containing soil resulted in high mortality (>65%). In contrast, the lower concentrations (500 and 1000 nematodes/pot) caused low mortality (35%) of C. hemipterus . Other heterorhabditid strains caused 95-100% mortality of C. humeralis in pot assay. The HP88 strain of H. bacteriophora and the Tx strain of Steinernema riobrave showed poor effectiveness. Incubation of different nematode strains with the C. humeralis larvae at high temperature (32 C) resulted in an increase in insect mortality with the IS-12 and IS-21 strains. Reduced mortality was recorded with the HP88 strain treatment at the higher temperature. The IS-5 and IS-12 strains were equally effective in all three soil types tested, whereas the IS-19 strain was more effective in the Almog type soil than in the others.     

Effectiveness of Entomopathogenic Nematodes in an Alginate Gel Formulation Against Lepidopterous Pests

An improved calcium alginate gel formulation was developed and tested as a carrier for entomopathogenic nematodes against Spodoptera littoralis and Helicoverpa armigera larvae. Mortality of 100% was caused in 4th instar larvae of the two insects by feeding them on 1000 infective juveniles (IJ) g -1 of Steinernema carpocapsae (ALL strain) in the gel for 24 h. Exposing 2nd to 5th instars of H. armigera and 3rd to 6th of S. littoralis to 500 IJ g -1 of S. carpocapsae (ALL strain) resulted in 70-100% larval mortality. Mature larvae were less susceptible to the nematodes. Mortality of larvae exposed to 500 IJg -1 of S. carpocapsae (ALL strain) ranged from about 45-55% at 4 h to 90-95% at 48 h. Fourth instar larvae fed for 24 h with 250 IJ g -1 of nematode strains in gel showed in S. littoralis ranges of susceptibility in the following descending order: S. feltiae (IS -7 strain) = S. carpocapsae (DT strain) = S. feltiae (IS-6 strain) > S. carpocapsae (Mexican strain) = S. carpocapsae (ALL strain) = Heterorhabditis bacteriophora (HP-88 strain) = H sp. (IS-5 strain) > S. riobravae (Texas strain); in H. armigera the rating was: S. feltiae (IS-7 strain) = H. bacteriophora (HP88 strain) > S. carpocapsae (ALL strain) = S. feltiae (IS-6 strain ) = Heterorhabditis sp. (IS5 strain) > S. carpocapsae (Mexican strain) > S. riobravae (Texas strain) . The number of nematodes per larval cadaver increased with mortality rates. In greenhouse tests at 28 &#45 2&#176;C and 90% relative humidity, gel discs containing 500 IJ g -1 of nematodes were pinned to leaves of potted plants of cotton ( Gossypium hirsutum ) (Acala SJ2) and the plants were offered to S. littoralis larvae. Larval mortality of 89 &#45 12.7% was caused by S. feltiae (IS-7 strain) and most of the plant leaves were protected against the larvae by the nematodes. In the control, larval mortality was 3.3 &#45 0.05% and the plants were almost completely defoliated. Possibilities of using the gel-nematode formulation to protect sheltered crops against insect pests are discussed     

Evaluation of the efficacy of the entomopathogenic nematodes Heterorhabditis sp. against sap beetles (Coleoptera: Nitidulidae)

Absract Nitidulid beetles (Coleoptera) are considered serious pests of date palms throughout the world. They attack the ripe fruit, causing it to rot, and damage is reflected in both reduced yield and lower fruit quality. Previous studies demonstrated the susceptibility of larvae of this pest to entomopathogenic nematodes from the genus Heterorhabditidis. In the present study nematode efficacy was evaluated in greenhouse and field. In containers filled with soil, moderate reduction in insect emergence was achieved when the nematodes were applied at concentrations of 25 and 50 IJs/cm². However, the highest concentration (100 IJs/cm²) treatment resulted in a drastic reduction (by 70–90%) in emergence of the beetles. The lowest emergence was achieved by the IS-19 and IS-21 strains (>10%). Efficacy of the IS-19 strain was retained up to 7 days after application at a rate of 100 IJs/cm². When the insect larvae were introduced to the soil 2 weeks after nematode application, the percentage emergence of insects increased by 2–2.5 fold as compared to previous introductions but was still lower than in the control. Insect density per container did not have an effect on efficacy of the nematodes when the strains IS-19 and IS-12 were used. Two field trials were conducted in different sites in Israel. In the first trail, conducted in date palm orchard, four strains of Heterorhabditis sp. were tested. No significant difference in insect emergence was recorded among the various treatments or the control. Whereas in the second trial conducted in a fig orchard, substantial reduction (by 50–70%) in insect emergence was recorded following nematode treatment. Further studies, under natural conditions, are needed to optimize application efficiency and evaluate the commercial utilization of these biological control agents.     

Screening Spanish isolates of steinernematid nematodes for use as biological control agents through laboratory and greenhouse microcosm studies

Entomopathogenic nematodes (EPNs) are one of the best non-chemical alternatives for insect pest control, with native EPN strains that are adapted to local conditions considered to be ideal candidates for regional biological control programs. Virulence screening of 17 native Mediterranean EPN strains was performed to select the most promising strain for regional insect pest control. Steinernema feltiae (Filipjev) (Rhabditida: Steinernematidae) Rioja strain produced 7%, 91% and 33% larval mortality for the insects Agriotes sordidus (Illiger) (Coleoptera: Elateridae), Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) and Ceratitis capitata (Wiedemann) (Diptera: Tephritidae), respectively, and was selected as the most promising strain. The S. feltiae Rioja strain-S. littoralis combination was considered the most suitable to develop the Rioja strain as a biocontrol agent for soil applications. The effect of soil texture on the virulence of the Rioja strain against S. littoralis was determined through dose-response experiments. The estimated LC₉₀ to kill larvae in two days was 220, 753 and 4178 IJs/cm² for soils with a clay content of 5%, 14% and 24%, respectively, which indicates that heavy soils produced negative effects on the virulence of the Rioja strain. The nematode dose corresponding to the LC₉₀ for soils with a 5% and 14% clay content reduced insect damage to Capsicum annuum Linnaeus (Solanales: Solanaceae) plants under greenhouse microcosm conditions. The results of this research suggest that an accurate characterization of new EPN strains to select the most suitable combination of insect, nematode and soil texture might provide valuable data to obtain successful biological control under different ecological scenarios in future field applications.     

Efficacy of entomopathogenic nematodes against potato tuber moth,Phthorimaea operculella (Lepidoptera: Gelechiidae) under laboratory conditions

The susceptibility of potato tuber moth, Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae) to native and commercial strains of entomopathogenic nematodes (EPNs) was studied under laboratory conditions. Native strains of EPNs were collected from northeastern Iran and characterised as Steinernema feltiae and Heterorhabditis bacteriophora (FUM 7) using classic methods as well as analysis of internal transcribed spacer (ITS) and D2/D3 sequences of 28S genes. Plate assays were performed to evaluate the efficiency of five EPN strains belonging to four species including Steinernema carpocapsae (commercial strain), S. feltiae, Steinernem glaseri and H. bacteriophora (FUM 7 and commercial strains). This initial assessment with 0, 75, 150, 250, 375 and 500 IJs/ml concentrations showed that S. carpocapsae and H. bacteriophora caused the highest mortality in both larval and prepupal stages of P. operculella, PTM. Thereafter, these three strains (i.e. S. carpocapsae, H. bacteriophora FUM 7 and the commercial strains) were selected for complementary assays to determine the effects of soil type (loamy, loamy–sandy and sandy) on the virulence of EPNs against the second (L2) and fourth instar (L4) larvae as well as prepupa. A soil column assay was conducted using 500 and 2000 IJs in 2-ml distilled water. Mortality in the L2 larvae was not affected by the EPN strain or soil type, while there was a significant interactive effect of nematode strains and soil type on larval mortality. The results also showed that EPN strains have higher efficiency in lighter soils and caused higher mortality on early larvae than that in loamy soil. In L4 larvae, mortality of PTM was significantly influenced by nematode strain and applied concentrations of infective juveniles. The larval mortality induced by S. carpocapsae was higher than those caused either by a commercial or the FUM 7 strain of H. bacteriophora. Prepupa were the most susceptible stage.     

Efficacy of indigenous entomopathogenic nematodes (Rhabditida: Heterorhabditidae, Steinernematidae), from Rio Grande do Sul Brazil, against Anastrephafraterculus (Wied.) (Diptera: Tephritidae) in peach orchards

Laboratory, greenhouse, and field experiments were performed with the objective of selecting efficient indigenous strains of entomopathogenic nematodes (EPNs) from Rio Grande do Sul (RS) state, Brazil, for controlling the South American fruit fly, Anastrepha fraterculus (Wied.). Laboratory experiments were conducted in 24 well-plates filled with sterile sand and one insect per well. In greenhouse experiments, plastic trays filled with soil collected from the field were used, while in field experiments, holes were made in soil under the edge of peach tree canopies. Among 19 EPN strains tested, Heterorhabditis bacteriophora Poinar RS88 and Steinernema riobrave Cabanillas, Poinar, & Raulston RS59 resulted in higher A. fraterculus larval (pre-pupal) and pupal mortality, with LD₉₀ of 1630, 457 and 2851, 423 infective juveniles (IJs)/cm², respectively. Greenhouse experiments showed no differences in pupal mortality at 250 and 500 IJs/cm² of either nematode. In the field, H. bacteriophora RS88 and S. riobravae RS59 sprayed individually over natural and artificially infested fruit (250 IJs/cm²) resulted in A. fraterculus larval mortality of 51.3%, 28.1% and 20%, 24.3%, respectively. There was no significant difference in A. fraterculus pupal mortality sprayed with an aqueous suspension of either nematode; however, when using infected insect cadavers, H. bacteriophora RS88 was more efficient than S. riobrave RS59. Our results showed that H. bacteriophora RS88 was more virulent to insect larvae, with an efficient host search inside the infested fruit and control of pupae in the soil after being applied by aqueous suspension or infected cadavers.     

The reproduction potentials of four entomopathogenic nematode strains related to cost-effective production for biological control

Bioassays to evaluate the mortality, virulence and reproduction potentials of four indigenous EPN strains, S-PQ16, S-BM12, H-KT3987 and H-CB3452 on insect larvae of mealworm (Tenebrio molitor) and greater wax moth (Galleria mellonella) revealed the highest mortality rates of two insect larvae at the highest inoculation dose of 100 IJs to range from 89 to 100 percent and 94.3–100 percent at 48 h after inoculation, respectively. Virulence was high for all nematode strains, with LC₅₀ values between 29.6 and 47.3 IJs/insect host. The highest IJ yields were different between nematode strains and insect host, from 66.8 × 10³ IJs (S-PQ16) to 118.6 × 10³ IJs (H-KT3987) on T. molitor, and from 54.2 × 10³ IJs (S-BM12) to 163.3 × 10³ IJs (H-KT3987) on G. mellonella. The culturing cost in terms of food expenditure for rearing insect larvae varied between insect larvae and nematode strains, from 6.76 to 26.63 USD per billion IJs for nematode strains cultured on T. molitor larvae and from 3.54 to 7.81 USD per billion IJs for nematode strains cultured on G. mellonella larvae. The full cost for a nematode product of 2.5 × 10⁹ IJs per hectare, produced through in vivo mass culturing, of the most efficient nematode strain, H-KT3987, was 191.3 USD, slightly cheaper than 199.4 USD for the same nematode product produced through in vitro mass culturing.     

Pathogenicity of Heterorhabditis nematodes isolated from north-western Himalaya against the larvae of Plutella xylostella (L.) (Lepidoptera: Plutellidae)

The efficacy of three entomopathogenic nematodes (Heterorhabditis spp.), from north western Himalaya, India was studied against the diamondback moth, Plutella xylostella (Linnaeus, 1758) (Lepidoptera: Plutellidae), under laboratory conditions. The larvae were exposed to 10, 20, 30 and 40 infective juveniles (IJs) of each nematode species for different time periods and they were found to be susceptible to all the EPNs tested. However, the susceptibility of larvae to nematode infection varied according to the dosages of IJs and their exposure periods. The efficacy of these indigenous entomopathogenic nematodes was also evaluated against the commercially available entomopathogenic nematode H. indica. An indigenous isolate, H. bacteriophora (HRJ), along with the commercial isolate H. indica recorded 100.0% mortality of insect larvae in 96 h exposure time against third instar larvae of P. xylostella. However, it was noticed that with the advancement of larval stage its mortality rate reduces and vice versa with the exposure period. All the tested nematode species were also found to reproduce within the host and produced infective juveniles. In conclusion, the evidence obtained in this study suggests that all the three indigenous EPN species are virulent enough to produce 100% mortality of larvae of P. xylostella. These EPN species thus have potential for the management of P. xylostella under integrated management practices.     

Entomopathogenic nematodes as potential biological control agents of Popillia japonica (Coleoptera,Scarabaeidae) in Piedmont Region (Italy)

In 2014, the Japanese beetle Popillia japonica (Coleoptera: Scarabaeidae) was first spotted in northern Italy in the Nature Park of the Ticino Valley, its first detection in continental Europe. This polyphagous invasive species has the potential to cause serious losses to horticulture and agriculture. Particularly for its management in a Nature Park, environmentally friendly strategies are necessary. To develop baseline data for a biological control approach to the Italian outbreak of P. japonica, we conducted laboratory and field experiments testing several indigenous and commercial strains of the entomopathogenic nematode (EPN) species Heterorhabditis bacteriophora and Steinernema carpocapsae against P. japonica larvae. In the laboratory, strains of H. bacteriophora caused greater mortality (ranging from 57% to 100%) than those of S. carpocapsae (3% to 77%). In micro‐plot field tests carried out at three different times, the most virulent indigenous EPN strain, H. bacteriophora ItH‐LU1, showed again the best results ranging from 44% to 93% against young larvae. Finally, in a large‐plot field trial, the commercial H. bacteriophora product provided 46% larval mortality. This study shows that H. bacteriophora strains have good potential as biological control agents of larvae of the invasive P. japonica in northern Italy.     

Pathogenicity of Metarhizium anisopliae for Ceratitis capitata (Wied.) (Diptera: Tephritidae) in soil with different pesticides

This research intended to investigate if the presence of pesticides in the soil could affect the pathogenicity of Metarhizium anisopliae Metsch. (Sorokin) for Ceratitis capitata (Wied.) and assess the effect of conidia application as suspension or dry conidia. The fungicides chlorothalonyl and tebuconazol, the acaricide abamectin, the insecticide trichlorfon, and the herbicide ametrin were applied at the manufacturer-recommended doses. Soil samples were placed in glass flasks and were given the fungus as conidial suspension or dry. After pesticide application, 20 3rd-instar larvae were placed in the soil. The flasks were sealed with voile fabric and incubated at 27 +/- 0.5 masculineC for nine days, until adult emergence; incubation continued for four more days at room temperature. The total insect survival was significantly affected and pathogenic activity was detected from the pupa stage on. Pupa survival was reduced (P<0.05); the same occurred during the adult phase. No effect was observed at the larval stage. The pesticides applied to the soil affected the activity of M. anisopliae slightly: only in the dry conidia assay the fungicides chlorothalonyl and tebuconazole reduced (86.2% and 82.5%, respectively) the survival period of C. capitata compared to the control (95.0%). The techniques used for conidia application did not influence the total insect survival rate, but conidial suspension applied on soil surface reduced survival during the pupae and adult phases.     

Effects of Soil Moisture on Control of Heterodera schachtii with Aldicarb

Soil moisture and the nematode population density in aldicarb-treated soil influenced control of the sugarbeet nematode, Heterodera schachtii. Greater numbers of nematode larvae infected 14-day-old sugarbeet seedlings growing in aldicarb-treated soil at 20-30% than at 80-100% field capacity (F. C.), and plant growth was inversely related to nematode infection and the nematode population density. Compared with that of control plants, plant growth increase also was greater at 80-100% F. C. when the nematode population was above 1.8 larvae/gm soil. A nematode population of 1.8 larvae/gm soil did not significantly affect sugarbeet yields. Aldicarb gave less control when soil moisture levels dropped to 20 and 50% F. C. at nematode populations of 3.5 and 6.2 larvae/gm soil. More effective control was obtained wth soil moisture levels at or above 80% F. C. This difference was attributed to continued activity of the toxicant in the rhizosphere at the high moisture level.     

Assessing the Potential of Entomopathogenic Nematodes to Control the Grape Root Borer Vitacea polistiformis (Lepidoptera: Sesiidae) Through Laboratory and Greenhouse Bioassays

Seventeen entomopathogenic nematode species and strains were evaluated for virulence to the grape root borer, Vitacea polistiformis (Harris) in laboratory and greenhouse bioassays. Heterohabditis bacteriophora strain GPS11 and H. zealandica strain X1 produced a larval mortality rate of over 85% of larvae embedded in the root cambium in laboratory bioassays. The nematode species H. marelata and H. bacteriophora strain Oswego produced mortality rates of over 75%. Of the Steinernema species tested, S. carpocapsae strain 'All' performed the best with a mortality rate of 69%. All other nematode species and strains tested, with the exception of S. bicornutum , produced some degree of larval mortality. In the greenhouse bioassays, 93% control was achieved with H. zealandica strain X1 applied at 4 ×109 infective juveniles (IJs) acre1 -1 (9.88 ×10 9 IJs ha -1 ). H. bacteriophora strain GPS11 successfully reproduced in grape root borer larvae. The numbers of IJs produced within infected larvae were related to larval size. The survival rate of neonate larvae on grape root sections was 61%, which thus provides a means to rear the neonate larvae for bioassays.     

Biocontrol potential of the entomopathogenic nematodes Heterorhabditis bacteriophora and Steinernema carpocapsae on cucurbit fly,Dacus ciliatus (Diptera: Tephritidae)

Entomopathogenic nematodes (EPNs) from the families Steinernematidae and Hererorhabditidae are considered excellent biological control agents against many insects that damage the roots of crops. In a regional survey, native EPNs were isolated, and laboratory and greenhouse experiments were conducted to determine the infectivity of EPNs against the cucurbit fly, Dacus ciliatus Loew (Diptera: Tephritidae). Preliminary experiments showed high virulence by a native strain of Heterorhabditis bacteriophora Poinar (Rhabditida: Heterorhabditidae) and a commercial strain of Steinernema carpocapsae Weiser (Rhabditida: Steinernematidae). These two strains were employed for further analysis while another native species, Steinernema feltiae, was excluded due to low virulence. In laboratory experiments, larvae and adult flies were susceptible to nematode infection, but both nematode species induced low mortality on pupae. S. carpocapsae had a significantly lower LC₅₀ value against larvae than H. bacteriophora in filter paper assays. Both species of EPNs were effective against adult flies but S. carpocapsae caused higher adult mortality. When EPN species were applied to naturally infested fruit (150 and 300 IJs/cm²), the mortality rates of D. ciliatus larvae were 28% for S. carpocapsae and 12% for H. bacteriophora. Both EPN strains successfully reproduced and emerged from larvae of D. ciliates. In a greenhouse experiment, H. bacteriophora and S. carpocapsae had similar effects on fly larvae. Higher rates of larval mortality were observed in sandy loam and sand soils than in clay loam. The efficacy of S. carpocapsae and H. bacteriophora was higher at 25 and 30°C than at 19°C. The results indicated that S. carpocapsae had the best potential as a biocontrol agent of D. ciliatus, based on its higher virulence and better ability to locate the fly larvae within infected fruits.     

Digestive enzymes during development of Ceratitis capitata (Diptera:Tephritidae) and effects of SBTI on its digestive serine proteinase targets

The digestive system of Ceratitis capitata was characterized during its larval development and in the insect stage. Disaccharidases against maltose and sucrose were more evident in the 2nd and 3rd day of larval development and in the adult stage, respectively. Glycosil-hydrolyses with higher specific alpha-galactosidasic and beta-galactosidasic activities were detected in the 2nd and 3rd day of the larval stage, respectively. Specific proteolytic activities against azocasein showed an increase in the 4th and 5th day of the larval stage and in the adult stage. Specific hemoglobin activities were constant between 2nd and 6th day of the larval stage. The larvae used mainly serine proteinases, such as trypsin/chymotrypsin, and the adult insects only chymotrypsin-like enzymes in their digestive process. Two serine proteinases were separated from zymogram between the 4th and 5th day of larval development and in the adult stage. Effect of soybean trypsin inhibitor (SBTI, a serine proteinase inhibitor) on development of C. capitata was examined by bioassay. C. capitata was susceptible to SBTI which affected larval mass at ED50 3.01%.     

Efficacy of entomopathogenic nematodes against soil-dwelling life stages of western flower thrips, Frankliniella occidentalis (Thysanoptera: Thripidae).

The efficacy of six entomopathogenic nematode (EPN) strains was tested in a laboratory study against soil-dwelling life stages of western flower thrips (WFT), Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae). The EPN strain collections screened included two Heterorhabditis bacteriophora species, i.e., H. bacteriophora HK3 (H.b H) and H. bacteriophora HB Brecan (H.b B), three Steinernema feltiae species, i.e., S. feltiae Sylt (S.f S), S. feltiae OBSIII (S.f O), and S. feltiae strain CR (S.f C), and the S. carpocapsae strain DD136 (S.c D). All soil-dwelling life stages of WFT were susceptible to the tested EPN strains. The most virulent strains were S.f S, S.c D, and H.b H. The S.f O strain was highly virulent against late second instar larvae and prepupae of WFT under high soil moisture conditions, but less effective against pupae under comparatively drier soil conditions. Results from dose rate experiments indicate that a comparatively high concentration of 400 infective juveniles (IJs) per cm(2) was needed to obtain high mortality in all soil-dwelling life stages of WFT. However, dose rates of 100-200 IJs/cm(2) already caused 30-50% mortality in WFT. The chances for combining EPNs with other biological control agents of WFT are discussed.     

The nematode Neoaplectana carpocapsae and the beet armyworm Spodoptera exigua: Infectivity of prepupae and pupae in soil and of adults during emergence from soil

The nematode, Neoaplectana carpocapsae, infected >90% of the prepupae of Spodoptera exigua in soil even at concentrations as low as five nematodes/cm² of soil surface. Pupae were less susceptible to nematode infection in soil than prepupae, with mortality ranging from 10 to 24% and 10 to 83% for pupae exposed 3–5 days and 6–8 days to the nematode, respectively. Longer exposure (6–8 days) of the pupae to the nematode resulted in higher mortality with a positive relationship with increasing concentrations. Adults of S. exigua were susceptible to nematode infections as they emerged from the soil. The higher nematode concentrations (25 and 50 nematodes/cm²) resulted in higher adult mortality. The majority of nematode-induced mortality occurred within 24 hr after emergence. The susceptibility of emerging S. exigua adults to N. carpocapsae offers a new dimension for insect control.     

Diapause induction as an interplay between seasonal token stimuli,and modifying and directly limiting factors: hibernation in Chymomyza costata

Larvae of wild type (WT) strain of Chymomyza costata Zetterstedt (Diptera: Drosophilidae) enter diapause (stop developing) in response to short‐day signal at a constant 18 °C, whereas larvae of a non‐photoperiodic‐diapause (NPD) strain do not respond to photoperiodic signalling and continue in larval development irrespective of daylength. The present study shows that WT larvae also respond reliably to thermoperiodic signalling (daily cycles of temperature) under constant darkness, whereas the NPD larvae do not, suggesting that the pathways transducing the environmental token stimuli (photoperiod and thermoperiod) onto the diapause developmental programme might merge functionally in the central biological clock system known to be mutated in NPD strain. Temperature and larval population density modify the output of token stimuli signalling. High temperatures (>24 °C) tend to avert, whereas low temperatures (<18 °C), especially in combination with constant darkness, stimulate diapause induction in WT strain. Overcrowding (>200 larvae per 5 g of larval diet) lengthens the duration of larval development and induces a ‘diapause‐like’ developmental arrest of relatively weak intensity in up to 60% of larvae of both strains. At high temperatures (>30 °C), all WT larvae continue direct development but subsequently die during the pupal stage. Low temperature exposure (<12 °C) causes quiescence in the majority of the larvae of both strains. Starvation blocks development and causes mortality when applied in larvae younger than day 3 of the third instar. Older larvae survive starvation and their photoperiodically‐induced developmental pre‐programming is not affected. Collectively, the results show that diapause induction in C. costata is a result of various interacting effects of multiple environmental factors.     

Effects of Crop Residue on the Persistence of Steinernema carpocapsae

We determined the effects of crop residue on the persistence of an entomopathogenic nematode, Steinernema carpocapsae. During 2 consecutive years, nematodes were applied at rates of 2.5 × 10₄ and 1.0 × 10⁵ infective juveniles/m² to small field plots planted with corn. Nematode persistence was monitored by exposing Galleria mellonella larvae to soil samples from plots with and without crop residue (approximately 75% coverage of soybean stubble). Persistence of S. carpocapsae was significantly greater in crop residue plots than in plots without residue. In crop residue plots that received the higher rate of nematode application, larval mortality did not significantly decrease during the study period (3 to 5 days) and remained above 85%. In nematode-treated plots without crop residue, however, larval mortality fell from over 96% to below 11% and 35% in the first and second trials, respectively. The increased crop residue may have benefited nematode persistence through protection from desiccation or ultraviolet light. We conclude that increased ground cover in cropping systems (e.g., due to reduced tillage) may lead to increased insect pest suppression with entomopathogenic nematodes.     

Influence of soil temperature and moisture on the infectivity of entomopathogenic nematodes (Rhabditida: Heterorhabditidae, Steinernematidae) against larvae of Ceratitis capitata (Wiedemann) (Diptera: Tephritidae)

The Mediterranean fruit fly, Ceratitis capitata (Wiedemann), is considered one of the main pests that affect fruit production in the world. This insect spends part of its life cycle in the soil, making it a target for entomopathogenic nematodes. This work aimed at evaluating the influence of soil temperature and moisture on the infectivity of Heterorhabditis sp. RSC01 and Steinernema carpocapsae ALL to third-instars of C. capitata, and to compare the efficiency of these isolates at five different soil temperatures (19, 22, 25, 28, and 31°C) and three levels of relative soil moisture (100, 75, and 50% of field capacity). Ten C. capitata larvae were transferred to plastic jars (12 cm × 6 cm) containing 100 g soil, followed by the application of an aqueous suspension containing 125 infective juveniles (IJ)/cm2. In the control treatment, 3 ml of distilled water was applied. Mortality evaluations were made five days later and were confirmed by observations of the characteristic symptoms and cadaver dissection. The infectivity was directly proportional to temperature increase, with maximum percent mortality of 86.7% and 80.0% for S. carpocapsae and Heterorhabditis sp., respectively, at 31°C. At 25°C, the highest mortality for both species was obtained at 75% of field capacity (96.7% and 26.7% for S. carpocapsae and Heterorhabditis sp., respectively).     

Investigations on the parasitism ofOtiorrhynchus salicicola andO. Sulcatus [Col.: Curculionidae] byHeterorhabditis sp. [Nematoda]

Experiments were conducted to study the efficacy of the insect parasitic nematodeHeterorhabditis sp. (HW79) as a biological control agent ofOtiorrhynchus salicicola. This weevil species is reported as a pest of ornamental plants in Switzerland and Italy. Dipping plastic boxes containing heavily infested cuttings of laurel (Prunus laurocerasus) in a nematode suspension resulted in approximately 100% parasitisation of full-grown larvae, pupae and non-emerged young adults. The average dose resulting from dipping varied between 56,000 and 62,000 nematodes per liter soil. This experiment was run under natural outdoor conditions. In a further outdoor experiment, pottedLigustrum plants were inoculated with eggs ofO. salicicola and later 20,000 infective juvenile nematodes per liter soil were added to the soil surface. The resulting weevil mortality in the treated pots was 78%. In seven greenhouse tests using the same nematode dose in pots with horticultural soil to which weevil larvae had been added, weevil mortality varied between 76% and 100%, the arithmetic average being 90%. These results indicate that Heterorhabditid nematodes may provide an effective means of controllingO. salicicola. In an other experiment usingO. sulcatus larvae, the influence of application time on nematode efficacy was investigated. When nematodes were added a few days before weevil larvae had hatched from the eggs, no parasitic effect was obtained. Nematode applications done shortly after larval hatching however, resulted in complete weevil control. These results are of significance in timing nematode applications in practice.