Evaluation of Entomopathogenic Nematodes for the Control of Mediterranean Fruit Fly (Diptera: Tephritidae)

The virulence of various entomopathogenic nematode (EPN) strains was evaluated against the Mediterranean fruit fly, C. capitata . The selected nematodes were assessed for their infectivity for the final larval stage of the insect host and under varying environmental conditions. Among 12 EPN strains tested, Steinernema riobrave Texas ( Sr TX) and Heterorhabditis sp. IS-5 (H IS-5), showed high activity and induced >80% mortality. Six EPN strains showed limited activity (>30% mortality), and four strains had no effect (<20% mortality). Sr TX was more effective than H IS-5. Mature C. capitata larvae were most susceptible to nematode infection during the first 4h after they began to emerge from their diet to pupate. Activity of the two nematode strains at a constant inoculation rate was dependent on insect larval density. The highest activity was recorded at 1.88 larvae cm -2 and decreased at higher larval densities. EPN activity was also directly related to nematode density. Maximal activity was shown at a density of 150 infective juveniles cm -2 . A similar activity pattern was also recorded with Sr TX in four different soil types. The persistence of this EPN in the soil extended over 5 days but there was no activity after 14 days. Except for a lower activity under cool conditions (17°C), temperatures ranging between 22 and 41°C, or moisture levels in the treated soil ranging between 3 and 20%, had no significant effect on nematode activity. Our results suggest that application of Sr TX against C. capitata may have potential for controlling C. capitata .     

Mortality rate of adult ticks due to infection by entomopathogenic nematodes

The mortality of Boophilus annulatus, Hyalomma excavatum, Rhipicephalus bursa, and Rhipicephalus sanguineus adult ticks was recorded after their exposure in petri dishes to 5 entomopathogenic nematode strains. The strains used were DT and Mexican of Steinernema carpocapsae, Hb HP88 of Heterorhabditis bacteriophora, and IS-3 and IS-5 of Heterorhabditis sp. The most rapid killing rates (LT50 and LT90) were observed for B. annulatus ticks (0.8-5.0 days). Most unfed males died 0.3-2.8 days after the unfed females, whereas even more time (0.8-8.0 days) passed before engorged females died. In most bioassays, the IS-3 and IS-5 strains of Heterorhabditis sp. nematodes appear to be the most pathogenic for ticks and, in most cases, killed ticks several days before the other 3 nematode strains. Unfed adult ticks exposed to nematodes died within less time than it takes for adults to complete their prefeeding period after molting.     

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     

Virulence of entomopathogenic nematodes to pecan weevil larvae, Curculio caryae (Coleoptera: Curculionidae), in the laboratory

The pecan weevil, Curculio caryae (Horn), is a key pest of pecans in the Southeast. Entomopathogenic nematodes have been shown to be pathogenic toward the larval stage of this pest. Before this research, only three species of nematodes had been tested against pecan weevil larvae. In this study, the virulence of the following nine species and 15 strains of nematodes toward fourth-instar pecan weevil was tested: Heterorhabditis bacteriophora Poinar (Baine, HP88, Oswego, NJ1, and Tf strains), H. indica Poinar, Karunakar & David (original and Homl strains), H. marelatus Liu & Berry (IN and Point Reyes strains), H. megidis Poinar, Jackson & Klein (UK211 strain), H. zealandica Poinar (NZH3 strain), Steinernema riobrave Cabanillas, Poinar & Raulston (355 strain), S. carpocapsae (Weiser) (All strain), S. feltiae (Filipjev) (SN strain), and S. glaseri (Steiner) (NJ43 strain). No significant difference in virulence was detected among nematode species or strains. Nematode-induced mortality was not significantly greater than control mortality (in any of the experiments conducted) for the following nematodes: H. bacteriophora (Baine), H. zealandica (NZH3), S. carpocapsae (All), S. feltiae (SN), S. glaseri (NJ43), and S. riobrave (355). All other nematodes caused greater mortality than the control in at least one experiment. Heterorhabditis megidis (UK211) but not H. indica (original) displayed a positive linear relationship between nematode concentration and larval mortality. Results suggested that, as pecan weevil larvae age, they may have become more resistant to infection with entomopathogenic nematodes.     

Virulence of entomopathogenic nematodes to Diaprepes abbreviatus (Coleoptera: Curculionidae) in the laboratory

The Diaprepes root weevil, Diaprepes abbreviatus (L.) is the most severe weevil pest in Florida citrus. Entomopathogenic nematodes have effectively suppressed larval populations of D. abbreviatus. Our objective was to conduct a broad laboratory comparison of entomopathogenic nematodes for virulence toward larvae of D. abbreviatus. The study was conducted at three temperatures (20, 24, and 29 degrees C) and included nine entomopathogenic species and 17 strains: Heterorhabditis bacteriophora Poinar (Baine, NJl, Hb, Hbl, HP88, and Lewiston strains), H. indica Poinar, Karunakar & David (original and Homl strains), H. marelatus Liu & Berry (IN and Point Reyes strains), H. megidis Poinar, Jackson & Klein (UK21l strain), H. zealandica Poinar (NZH3 strain), Steinernema riobrave Cabanillas, Poinar & Raulston (355 strain), S. carpocapsae (Weiser) (All strain), S. feltiae (Filipjev) (SN and UK76 strains), and S. glaseri (Steiner) (NJ43 strain). At 20 degrees C, the greatest mortality was caused by S. riobrave although it was not significantly greater than H. bacteriophora (Baine), H. bacteriophora (Hb), H. bacteriophora (Hbl), and H. indica (original). At 24 and 29 degrees C, S. riobrave caused greater larval mortality than other nematodes tested. Two strains of H. indica, H. bacteriophora (Baine), and S. glaseri were next in terms of virulence at 29 degrees C. Our results suggest that S. riobrave has the greatest potential for control of D. abbreviatus.     

Parasitism of subterranean termites (Isoptera: Rhinotermitidae: Termitidae) by entomopathogenic nematodes (Rhabditida: Steinernematidae; Heterorhabditidae)

In laboratory bioassays, Steinernema riobrave Cabanillas, Poinar and Raulston (355 strain), Steinernema carpocapsae (Weiser) (Mexican 33 strain), Steinernemafeltiae (Filipjev) (UK76 strain), and Heterorhabditis bacteriophora Poinar (HP88 strain) were all capable of infecting and killing three termite species, Heterotermes aureus (Snyder), Gnathamitermes perplexus (Banks), and Reticulitermes flavipes (Kollar) in laboratory sand assays. S. riobrave and S. feltiae caused low levels of Reticulitermes virginicus (Banks) mortality under the same conditions. At 22 degrees C, significant mortality (> or = 80%) of worker H. aureus and G. perplexus was caused by S. riobrave, in sand assays, indicating the need for further study. Because of the short assay time (3 d maximum), reproduction of the nematodes in the target host species was not recorded. All nematode species were observed to develop to fourth-stage juveniles, preadult stages, or adults in all termite species with the exception of R. virginicus. Only S. riobrave developed in R. virginicus. Nematode concentration and incubation time had significant effects on the mortality of worker H. aureus. S. riobrave consistently generated the highest infection levels and mortality of H. aureus in sand assays.     

Biological Control of the Pecan Weevil,Curculio caryae (Coleoptera: Curculionidae), with Entomopathogenic Nematodes

Steinernema carpocapsae (Weiser) strain A11, S. feltiae (Filipjev) strain SN, and Heterorhabditis bacteriophora Poinar strains HP88 and Georgia were tested for their efficacy as biological control agents of the pecan weevil, Curculio caryae (Horn), in pecan orchard soil-profile containers under greenhouse conditions. Percentage C. caryae parasitism by S. carpocapsae and H. bacteriophora strain HP88 and Georgia was consistently poor when applied either prior to or following C. caryae entry into the soil, suggesting that these nematode species and (or) their enterobacteria are poor biological control agents of weevil larvae. Soil taken 21 days following application of S. carpocapsae or H. bacteriophora strain HP88 induced a low rate of infection of Galleria mellonella larvae, whereas soil that had been similarily treated with H. bacteriophora strain Georgia induced a moderate rate of infection. Percentage C. caryae parasitism by S. feltiae was consistently low when applied following C. caryae entry into the soil and was inconsistent when applied as a barrier prior to entry of weevil larvae into the soil. Soil taken 21 days following application of S. feltiae induced a high rate of infection of G. mellonella larvae.     

Environmental factors affecting sexual differentiation in the entomopathogenic nematode Heterorhabditis bacteriophora

The present study was aimed at determining the influence of various environmental factors on sex differentiation (SD) in the entomopathogenic nematode Heterorhabditis bacteriophora HP88 strain, under in vivo and in vitro culture conditions. Injection of individual nematodes into last instars of Galleria mellonella resulted in development of a similar number of females and hermaphrodites (35-40%) and 20-25% males. Increasing the number of nematodes injected into the insect did not change these proportions. In smaller insects (0.7-1.5 cm long), an increase in the proportion of hermaphrodites was recorded as compared with larger size cadavers (2.4-2.7 cm long). When individual hermaphrodites were placed on NGM, the proportion of hermaphrodites, females and male progeny was 63%, 31%, and 6%, respectively. Rearing on richer medium ("Dog-food" agar) resulted in reduction in the proportion of hermaphrodites. Nematodes introduced to the symbiotic bacterium obtained from other nematode strains (IS-5 and IS-33) developed similarly to the culture reared on the HP88 bacteria. Rearing the nematodes at a temperature range between 21 degrees C to 30 degrees C also did not have a significant effect on the sexual differentiation among nematodes cultured on NGM. The proportion of hermaphrodites increased as the starvation period of hatching nematode juveniles lengthened (>6 hr). The data obtained in the present study strongly suggest that the main factor affecting sex differentiation in H. bacteriophora is the nutrition source. The practical and biological implications of the results are discussed.     

Susceptibility of the boll weevil to Steinernema riobrave and other entomopathogenic nematodes

The susceptibility of the boll weevil (BW), Anthonomus grandis Boheman, to Steinernema riobrave and other nematode species in petri dishes, soil (Hidalgo sandy clay loam), and cotton bolls and squares was investigated. Third instar weevils were susceptible to entomopathogenic nematode (EN) species and strains in petri dish bioassays at 30 degrees C. Lower LC(50)'s occurred with S. riobrave TX- 355 (2 nematodes per weevil), S. glaseri NC (3), Heterorhabditis indicus HOM-1 (5), and H. bacteriophora HbL (7) than H. bacteriophora IN (13), S. riobrave TX (14), and H. bacteriophora HP88 (21). When infective juveniles (IJs) of S. riobrave were applied to weevils on filter paper at 25 degrees C, the LC(50) of S. riobrave TX for first, second, and third instars, pupae, and 1-day-old and 10-days-old adult weevils were 4, 5, 4, 12, 13, and 11IJs per weevil, respectively. The mean time to death, from lowest to highest concentration, for the first instar (2.07 and 1.27days) and second instar (2.55 and 1.39days) weevils were faster than older weevil stages. But, at concentrations of 50 and 100IJs/weevil, the mean time to death for the third instar, pupa and adult weevils were similar (1.84 and 2.67days). One hundred percent weevil mortality (all weevil stages) occurred 3days after exposure to 100IJs per weevil. Invasion efficiency rankings for nematode concentration were inconsistent and changed with weevil stage from 15 to 100% when weevils were exposed to 100 and 1IJs/weevil, respectively. However, there was a consistent relationship between male:female nematode sex ratio (1:1.6) and nematode concentration in all infected weevil stages. Nematode production per weevil cadaver increased with increased nematode concentrations. The overall mean yield of nematodes per weevil was 7680IJs. In potted soil experiments (30 degrees C), nematode concentration and soil moisture greatly influenced the nematode efficacy. At the most effective concentrations of 200,000 and 400,000IJs/m(2) in buried bolls or squares, higher insect mortalities resulted in pots with 20% soil moisture either in bolls (94 and 97% parasitism) or squares (92 and 100% parasitism) than those of 10% soil moisture in bolls (44 and 58% parasitism) or squares (0 and 13% parasitism). Similar results were obtained when nematodes were sprayed on the bolls and squares on the soil surface. This paper presents the first data on the efficacy of S. riobrave against the boll weevil, establishes the potential of EN to control the BW inside abscised squares and bolls that lay on the ground or buried in the soil.     

A Novel Strain of Steinernema riobrave (Rhabditida: Steinernematidae) Possesses Superior Virulence to Subterranean Termites (Isoptera: Rhinotermitidae)

Subterranean termites are major global pests of wood structures and wood products. Among the most economically important subterranean termite species in the US are Heterotermes aureus, Reticulitermes flavipes, and Coptotermes formosanus. In prior studies, the entomopathogenic nematode, Steinernema riobrave strain 355, exhibited a high level of virulence to H. aureus compared with other nematode species. However, S. riobrave 355 was reported to be poorly or only moderately virulent to R. flavipes and C. formosanus, respectively. We hypothesized that other strains of S. riobrave may possess a high level of virulence to all three termite species. Under laboratory conditions we compared three novel strains of S. riobrave (3-8b, 7-12, and TP) with the 355 strain for virulence to H. aureus, R. flavipes, and C. formosanus workers. H. aureus was very susceptible to all the S. riobrave strains, and termites in all nematode treatments were dead after 4 d. The TP strain of S. riobrave caused greater mortality in R. flavipes and C. formosanus compared to the other nematode strains. Specifically, the TP strain caused 75% and 91% mortality in R. flavipes and C. formosanus, respectively, which was more than 300% and 70% higher than the mortality caused by other strains. Additional studies are warranted to determine the ability of S. riobrave (TP) to control the targeted termite species under field conditions.     

Evaluating mulches together with Heterorhabditis zealandica (Rhabditida: Heterorhabditidae) for the control of diapausing codling moth larvae,Cydia pomonella (L.) (Lepidoptera: Tortricidae)

The potential of using an entomopathogenic nematode, Heterorhabditis zealandica Poinar, together with different test mulches (pine chips, wheat straw, pine wood shavings, blackwood and apple wood chips) to control diapausing codling moth, Cydia pomonella (L.) larvae was evaluated. Mesh cages were identified as a suitable larval-containment method. High levels of codling moth mortality were obtained when using pine wood shavings as mulch (88%) compared to pine chips, wheat straw, blackwood and apple wood chips (41–88%). Humidity (>95% RH) has to be maintained for at least 3 days to ensure nematode survival. It was also proven that nematodes had the ability to move out of infected soil into moist mulch, to infect the codling moth larvae residing at heights of up to 10 cm. Field experiments showed the importance of climatic conditions on nematode performance. Low temperatures (<15°C) recorded during the first trial resulted in low levels of control (48%), as opposed to the 67% mortality recorded during the second trial (temperatures ranged between 20 and 25°C). Low levels of persistence (<10%) were recorded in the mulches post-application. The study conclusively illustrated some of the baseline requirements fundamental to the success of entomopathogenic nematodes together with mulches for the control of codling moth.     

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.     

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.     

Field Efficacy of Entomopathogenic Nematodes against the Sugar-beet Weevil Temnorhinus ( = Conorrhynchus ) mendicus Gyll. (Coleoptera: Curculionidae)

In 1992 and 1993, the field effectiveness of Heterorhabditis sp. (NL-HL81 strain), H. bacteriophora (HP 88 strain) and Steinernema carpocapsae ('All' strain) against the larvae of Temnorhinus mendicus Gyll. was assessed. The biological tests were compared with two chemical treatments (cypermethrin or deltamethrin) and one untreated control. In 1992, S. carpocapsae gave better results than Heterorhabditis sp. in reducing the percentage of infested roots, as compared with the untreated sample and the chemical one; similarly, the irrigated control gave the best results. In 1993, three concentrations of entomopathogenic nematodes (EPNs) were tested: 0.250 106 infective juveniles (IJs) m - 2, 0.125 106 IJs m - 2 and 0.075 106 IJs m - 2. The different numbers of EPNs did not give very different results from each other; however, H. bacteriophora at 0.075 106 IJs m - 2 was the least effective. In general, cypermethrin was more effective than deltamethrin, but one treatment with EPNs followed by irrigation was always more effective than two chemical applications.     

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.     

Evaluation of certain entomopathogenic nematodes against the melon ladybird,Epilachna chrysomelina F. (Coleoptera: Coccinellidae)

Three native Egyptian nematode isolates; Heterorhabditis taysearae and Heterorhabditis sp. S1 (Heterorhabditidae) and Steinernema carpocapsae S2 (Steinernematidae) as well as H. bacteriophora Hp88 as an imported species, were used in the present work to evaluate their activities against larvae and adults of the melon ladybird, Epilachna chrysomelina. The target pest was found to be susceptible to all tested entomopathogenic nematodes under laboratory conditions of 30±5°C.

In the greenhouse, a single spray of nematode suspension (1000 infective juveniles per ml) of each of H. taysearae, H. bacteriophora Hp88 and Steinernema carpocapsae S2 on squash seedlings was enough to give a reasonable mortality of 4^th larval instar E. chrysomelina, reaching 65.2, 44.0 and 84.0%, respectively, one week after application. This gives evidence that the Egyptian nematode isolates could tolerate high temperature and could be recommended for application in the control programmes of E. chrysomelina larvae in cucurbit fields.     

Biological control potential of native entomopathogenic nematodes (Steinernematidae and Heterorhabditidae) against Spodoptera cilium (Lepidoptera: Noctuidae) in turfgrass

In laboratory studies, we demonstrated that five native entomopathogenic nematode species/isolates caused 100% mortality of Spodoptera cilium larvae, a soil surface-feeding pest of turfgrass. At 25 infective juveniles/cm² applied to sod, two selected Turkish species, Steinernema carpocapsae and Heterorhabditis bacteriophora (Sarigerme isolate), averaged 77% and 29% larval mortality, respectively.     

河北异小杆线虫一品系的分类鉴定及其对蛴螬致病力的测定

昆虫病原线虫是一种有应用潜力的地下害虫生物防治因子。本研究利用形态学特征和ITS-rDNA分析方法对从河北省沧州分离的异小杆线虫一品系进行了鉴定, 并在室内测定比较了其对蛴螬的致病力。通过对该线虫侵染期幼虫和雄性成虫主要形态学特征的参数测量, 发现其与嗜菌异小杆线虫Heterorhabditis bacteriophora的形态特征最为相近;同时ITS1-rDNA序列比对和系统发育学分析结果显示其与嗜菌异小杆线虫亲缘关系最近。结合形态学和分子生物学特征, 确定该线虫为嗜菌异小杆线虫沧州品系。该线虫对蛴螬（华北大黑鳃金龟Holotrichia oblita、暗黑鳃金龟H. parallela和铜绿丽金龟Anomala corpulenta 3种金龟子的2龄幼虫）的致病力研究结果表明：处理72 h后, 暗黑鳃金龟幼虫死亡率显著高于另外两种金龟子幼虫 (P<0.05);处理120 h后, 暗黑鳃金龟和铜绿丽金龟幼虫的死亡率分别达到93.3%和80.0%, 二者无显著差异 (P>0.05), 可见该线虫对它们有较强的致病力。不同线虫对暗黑鳃金龟幼虫的致病力结果显示, 嗜菌异小杆线虫沧州品系侵染96 h后, 幼虫的死亡率显著高于Steinernema feltiae和S. longicaudum两种线虫的处理 (P<0.05) , 说明嗜菌异小杆线虫沧州品系的致病力显著高于另外两种线虫。     

New entomopathogenic nematodes from semi-natural and small-holder farming habitats of Rwanda

Five field surveys for indigenous entomopathogenic nematodes (EPNs) were conducted in 22 semi-natural and 17 small-holder farming habitats across 16 districts of different altitudes in the northern, eastern, southern and Kigali city provinces of Rwanda. In 2014, 216 mixed soil samples were collected and subsamples thereof baited with Galleria mellonella or Tenebrio molitor larvae. Five samples from five locations and habitats were positive for nematodes (2.8%). Nine nematode species/strains were isolated and five successfully maintained. DNA sequence comparisons and morphological examinations revealed Steinernema carpocapsae, Heterorhabditis bacteriophora, as well as two steinernematids and one heterorhabditid with no species designation. The isolates (strains) were named Steinernema sp. RW14-M-C2a-3, Steinernema sp. RW14-M-C2b-1, Steinernema carpocapsae RW14-G-R3a-2, H. bacteriophora RW14-N-C4a and Heterorhabditis sp. RW14-K-Ca. These are the first records of naturally occurring EPNs in Rwanda. It is also the first record of S. carpocapsae from Africa. Finding H. bacteriophora from tropical rather than temperate Africa was surprising. The found nematodes will serve as the basis for efficacy screening, and for mass production in a biocontrol agent factory at Rubona Research Centre of the Rwanda Agriculture Board with the ultimate aim of delivering effective, safe and environmentally benign pest control for soil-inhabiting pests.     

Ecological characterization of entomopathogenic nematodes isolated in stone fruit orchard soils of Mediterranean areas

Entomopathogenic nematodes in the families Steinernematidae and Heterorhabditidae were isolated from stone-fruit orchards in two Mediterranean regions of Spain. A total of 630 soil samples (210 sites) from Catalonia and 90 soil samples (30 sites) from Murcia were evaluated resulting in 5.2% and 20% of the soils testing positive for nematodes, respectively. Ten steinernematid isolates and three heterorhabditid isolates were recovered using the Galleria mellonella baiting method. Based on morphometric data, molecular data, and cross-breeding experiments the nematode species were identified as Steinernemafeltiae and Heterorhabditis bacteriophora. Environmental tolerance to heat, desiccation and hypoxia, the effect of temperature on infectivity and reproduction and nematode migration in sand columns were compared among isolates and one Steinernema carpocapsae strain. Results showed differences among species and a great variability within species. Beneficial traits for each strain were added up to identify a superior candidate to control Mediterranean flat-headed rootborer, Capnodis tenebrionis. When all analyzed factors were considered, three S. feltiae isolates (Bpa, Sor and M116) obtained the best scores, and when hypoxia was removed, two of the strains (Bpa and Sor) continued ranking superior to other strains.