Key elements in the successful control of diapausing codling moth,Cydia pomonella (Lepidoptera: Tortricidae) in wooden fruit bins with a South African isolate of Heterorhabditis zealandica (Rhabditida: Heterorhabditidae)

The non-insecticidal control strategies currently being implemented in South African orchards for the control of codling moth, Cydia pomonella (L.) may be hampered by wooden fruit bins being infested with diapausing codling moth larvae, acting as a potential source of re-infestation. Key factors contributing to the success or failure of an entomopathogenic nematode application were investigated using the SF 41 isolate of Heterorhabditis zealandica in laboratory bioassays with wooden minibins. Under operational conditions, an application rate of 100 IJs/mL (LD₉₀₌102 IJs/mL) effectively controlled codling moth larvae in these bins, and for further laboratory bioassays, the LD₅₀ value of 18 IJs/mL (?25 IJs/mL) was identified as the discriminating dosage. Maximum mortality was attained when bins were pre-wet for at least 1 min (>90% RH) and maintained at maximum humidity (>95% RH) post-treatment for at least 3 days (LT₉₀=73 h), to ensure nematode survival and subsequent satisfactory infection of diapausing codling moth larvae. Tarping bins achieved the desired high level of humidity required. Furthermore, adjuvants (specifically Reverseal 10?) also improved an application. The study conclusively illustrated that if all the above-mentioned conditions are met, H. zealandica has the potential to successfully disinfest wooden fruit bins of codling moth.     

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.     

Entomopathogenic nematodes from north-eastern South Africa and their virulence against false codling moth,Thaumatotibia leucotreta (Lepidoptera: Tortricidae)

A survey was undertaken to determine the diversity and frequency of endemic entomopathogenic nematodes (EPN) in subtropical regions in the Mpumalanga, Limpopo and KwaZulu-Natal provinces of South Africa. A total of 136 soil samples were randomly taken from cultivated and uncultivated habitats, including subtropical fruit tree orchards (avocado, litchi, macadamia, mango and guava) and adjoining natural vegetation. EPNs were isolated from 14 samples (10.3%) by means of baiting with Tenebrio molitor larvae. Heterorhabditis was the most common genus isolated from 12 samples, whereas only two Steinernema species were isolated. The most common Heterorhabditis isolated were Heterorhabditis noenieputensis and H. zealandica, which were both isolated from four samples each. The present report is the first occurrence of H. taysearae and H. baujardi in South Africa. The two Steinernema species are both unknown species. Laboratory bioassays were conducted to determine the potential of seven native EPN isolates to control the false codling moth (Thaumatotibia leucotreta) (FCM). Last instar FCM larvae were found to be most susceptible to S. yirgalemense (157-C), H. zealandica (WS 23) and Steinernema litchii (WS9), when exposed to 50 IJs/50?µ? water for 48?h, causing 100%, 94.2% and 93.5% mortality, respectively.     

Isolation and identification of entomopathogenic nematodes from citrus orchards in South Africa and their biocontrol potential against false codling moth

A survey was conducted to determine the diversity and frequency of endemic entomopathogenic nematodes (EPN) in citrus orchards in the Western Cape, Eastern Cape and Mpumalanga provinces of South Africa. The main aim of the survey was to obtain nematodes as biological control agents against false codling moth (FCM), Thaumatotibia leucotreta, a key pest of citrus in South Africa. From a total of 202 samples, 35 (17%) tested positive for the presence of EPN. Of these, four isolates (11%) were found to be steinernematids, while 31 (89%) were heterorhabditids. Sequencing and characterisation of the internal transcribed spacer (ITS) region was used to identify all nematode isolates to species level. Morphometrics, morphology and biology of the infective juvenile (IJ) and the first-generation male were used to support molecular identification and characterisation. The Steinernema spp. identified were Steinernema khoisanae, Steinernema yirgalemense and Steinernema citrae. This is the first report of S. yirgalemense in South Africa, while for S. citrae it is the second new steinernematid to be identified from South Africa. Heterorhabditis species identified include Heterorhabditis bacteriophora, Heterorhabditis zealandica and an unknown species of Heterorhabditis. Laboratory bioassays, using 24-well bioassay disks, have shown isolates of all six species found during the survey, to be highly virulent against the last instar of FCM larvae. S. yirgalemense, at a concentration of 50 IJs/FCM larva caused 100% mortality and 74% at a concentration of 200 IJs/pupa. Using a sand bioassay, S. yirgalemense gave 93% control of cocooned pupae and emerging moths at a concentration of 20 IJs/cm². This is the first report on the potential use of EPN to control the soil-borne life stages of FCM, which includes larvae, pupae and emerging moths. It was shown that emerging moths were infected with nematodes, which may aid in control and dispersal.     

Effect of humidity and a superabsorbent polymer formulation on the efficacy of Heterorhabditis zealandica (Rhabditida: Heterorhabditidae) to control codling moth,Cydia pomonella (L.) (Lepidoptera: Tortricidae)

Adequate moisture levels are required for nematode survival and subsequent efficacy as entomopathogens. Formulation of nematodes aimed at aboveground applications may assist in maintaining such moisture levels. In this study, we report the effects of a superabsorbent polymer formulation, Zeba® on the performance of an entomopathogenic nematode, Heterorhabditis zealandica Poinar, for controlling diapausing codling moth, Cydia pomonella (L.) larvae in cryptic habitats on trees. Water activity (a_w-value) on bark was considered to be an indication of moisture levels on trees in cryptic habitats where codling moth larvae are known to occur, thereby influencing nematode efficacy. H. zealandica was only able to infect codling moth larvae at a_w≥0.92, with a_w50=0.94 and a_w90=0.96. Laboratory experiments in which nematode concentration was investigated indicated a positive linear relationship between the concentration of nematodes applied and the level of control obtained, with the highest level of mortality recorded at 80 IJs/larva, requiring at least 4 h of conditions conducive to nematode activity to ensure infectivity and subsequent efficacy. Further experimentation showed that the use of the Zeba formulation, together with the nematodes, improved the level of control obtained at 60% and 80% RH in the laboratory and that it also enhanced the survival and infection-ability of the nematodes in the field. The study conclusively illustrates that the tested formulation assisted in maintaining adequate moisture levels on the application substratum, as required for nematode survival and subsequent efficacy.     

Predation of codling moth Cydia pomonella L. by the Silvereye Zosterops lateralis (Latham)

Predation of cocooned larvae of codling moth Cydia pomonella by silvereyes Zosterops lateralis was studied in an apple orchard in Nelson, New Zealand. Apple logs with known larval densities were made available to the birds for known periods of time, either in cages or exposed in the apple orchard. The numbers of silvereyes and the natural predation of codling moth were recorded in the same orchard. Predation was density dependent. On caged logs with an initial high density of 32 larvae, 1.1 larvae were consumed per bird‐hour; in contrast, one larva was consumed per 34.5 bird‐hours at three larvae per log. A curvilinear relationship was demonstrated between larval density and the bird‐hours required for predation; this relationship was consistent with the known density dependence of silvereye predation of codling moth. A regression of the total annual winter bird predation of larvae in the orchard on bird numbers was significant. However, the density dependence of predation resulted in declining rates of predation over the winter as larval density declined; the first birds to arrive in the orchard benefitted from particularly high predation rates. As a consequence, fluctuations in bird numbers during the winter had only a secondary influence on predation rates. The numbers of silvereyes in the orchard showed no relationship to the density of the codling moth population present. This study confirmed the importance of silvereyes in the predation of codling moth and a functional, not numerical, rseponse of these birds to codling moth density.     

Entomopathogenic Nematodes for Control of Codling Moth,Cydia pomonella(Lepidoptera: Tortricidae): Effect of Nematode Species, Concentration, Temperature, and Humidity

The susceptibility of codling moth diapausing larvae to three entomopathogenic nematode species was assessed in the laboratory using a bioassay system that employed cocooned larvae within cardboard strips. The LC₅₀values forSteinernema carpocapsae, S. riobrave,andHeterorhabditis bacteriophorawere 4.7, 4.8, and 6.0 infective juveniles/cm², respectively. When a discriminating concentration of 10 infective juveniles/cm²of each of the three nematode species was evaluated at 15, 20, 25, and 30°C,S. carpocapsaewas the most effective nematode with mortalities ranging from 66 to 90%. Mortalities produced byS. riobraveandH. bacteriophoraat the four temperatures were 2–94 and 25–69%, respectively. Studies were also conducted to test infectivity at 10, 35, and 40°C. No mortality was produced by any of the nematode species at 10°C.S. riobravewas the most infective nematode at 35°C producing 68% mortality which was more than twice that observed forS. carpocapsaeorH. bacteriophora.Codling moth larvae treated with 10 infective juveniles/cm²ofS. carpocapsaeand kept in 95+% RH at 25°C for 0–24 h followed by incubation at 25–35% RH indicated that more than 3 h in high humidity was needed to attain 50% mortality. Trials ofS. carpocapsae, S. riobrave,andH. bacteriophoraat 50 infective juveniles/cm²against cocooned larvae on pear and apple logs resulted in reductions of codling moth adult emergence of 83, 31, and 43%, respectively, relative to control emergence. Trials of the three entomopathogenic nematodes at 50 infective juveniles/cm²against cocooned larvae in leaf litter resulted in 99 (S. carpocapsae), 80 (S. riobrave), and 83% (H. bacteriophora) mortality, respectively. Our results indicate good potential of entomopathogenic nematodes, especiallyS. carpocapsae,for codling moth control under a variety of environmental conditions.     

Diversity and distribution of entomopathogenic nematodes (Steinernematidae, Heterorhabditidae) in South Africa

A total of 1506 soil samples from different habitats in seven geographic regions of South Africa were evaluated for the presence of entomopathogenic nematodes (EPN). Nematodes were isolated from 5% of the samples. Among the steinernematids, four Steinernema sp. were recovered including Steinernema khoisanae and three new undescribed species. Although steinernematids were recovered from both humid subtropical and semiarid regions, this family accounted for 80% of EPN recovered from the semiarid climate zones characterised by sandy, acidic soils. Eight isolates of S. khoisanae were recovered from the Western Cape province. One of the new undescribed steinernematids (Steinernema sp. 1) was recovered only from the Free State and KwaZulu-Natal provinces where humid subtropical conditions prevail and soils are generally less acidic with higher clay content. A high level of adaptation, however, was noted with Steinernema sp. 2, which was recovered from a wide range of soil conditions and habitats ranging from semiarid (Western Cape province) to humid subtropical (KwaZulu-Natal province). A third undescribed steinernematid, Steinernema sp. 3, seemed better adapted to heavier soils with more than 80% of isolates recovered from fruit orchards in the Free State province. Heterorhabditis bacteriophora was the only heterorhabditid recovered during this survey. This species was particularly prevalent in four provinces ranging from humid subtropical to semiarid regions. Isolation of EPN directly from insect cadavers included Steinernema sp. 2 and one H. bacteriophora from an unidentified white grub (Scarabaeidae) cadaver (i.e., dual infection) and H. bacteriophora from the black vine weevil, Otiorhynchus sulcatus.     

Susceptibility of the Mediterranean fruit fly (Ceratitis capitata) and the Natal fruit fly (Ceratitis rosa) to entomopathogenic nematodes

The potential of entomopathogenic nematodes, Heterorhabditis bacteriophora, Heterorhabditis zealandica and Steinernema khoisanae, to infect pupariating larvae, pupae and adults of Ceratitis capitata and Ceratitis rosa was investigated in laboratory bioassays. Pupariating larvae and adult flies were susceptible to nematode infection, with no infection recorded for the pupae. Pupariating larvae of C. capitata were generally more susceptible to infection than those of C. rosa. Significantly more larvae of C. capitata were infected by H. bacteriophora. For C. rosa, highest infectivity of larvae was obtained with H. zealandica. In contrast, adults of both species were highly infected by S. khoisanae.     

Influence of humidity and a surfactant-polymer-formulation on the control potential of the entomopathogenic nematode <Emphasis Type="Italic">Steinernema feltiae</Emphasis> against diapausing codling moth larvae (<Emphasis Type="Italic">Cydia pomonella</Emphasis> L.) (Lepidoptera: Tortricidae)

The codling moth (Cydia pomonella L.) is a serious pest of pome fruit. Diapausing cocooned larvae overwinter in cryptic habitats in the soil or in the bark of infested trees. The entomopathogenic nematode Steinernema feltiae (Filipjev) (Rhabditida: Steinernematidae) is used to control diapausing codling moth larvae. The objective of this study was to define environmental conditions favouring the performance of the nematodes. Cocooned larvae were more susceptible than non-cocooned larvae. Susceptibility of pupae was low. To determine the influence of decreasing water activity (a_w-value) on the activity of the nematodes, mortality of codling moth larvae and Galleria mellonella L. were tested in sand-sodium-polyacrylate mixtures of variable water activity. S. feltiae was able to infect both insects at a_w-values >0.9. Cocooned larvae of both insects died at lower a_w-values than non-cocooned larvae. Mortality of cocooned larvae did not further increase after half an hour of exposure to nematodes, whereas the mortality of non-cocooned larvae increased with increasing exposure time. LC₅₀ and LC₉₀ considerably decrease with increasing RH. The negative influence of the relative humidity (macro environment) was less important than the effect of the water activity in the bark substrate (micro environment). The micro environment can be manipulated by applying S. feltiae with higher volumes of water. A surfactant-polymer-formulation significantly increased nematode efficacy and can buffer detrimental environmental effects.     

Control of diapausing codling moth,Cydia pomonella (Lepidoptera: Tortricidae) in wooden fruit bins,using entomopathogenic nematodes (Heterorhabditidae and Steinernematidae)

Stacked wooden fruit bins are frequent overwintering sites for overwintering diapausing codling moth larvae. Control strategies against the codling moth (Cydia pomonella) (Lepidoptera: Tortricidae) in South Africa have been hampered by the reinfestation of orchards from nearby stacked infested fruit bins and by the movement of infested bins between orchards. Worldwide, wooden fruit bins are systematically being replaced with plastic bins, however in South Africa this will not be accomplished in the near future. The objective of this study was to evaluate the potential of two recycled commercially available entomopathogenic nematode (EPN) species, Heterorhabditis bacteriophora and Steinernema feltiae, as well as of a local species, Steinernema yirgalemense, to disinfest miniature wooden fruit bins under controlled conditions in the laboratory. After dipping miniature bins loaded with codling moth larvae in a suspension of 25?IJs/mL of each of the three EPN species, under optimum conditions of temperature and humidity, the highest percentage of control was obtained using S. feltiae (75%). The addition of adjuvants significantly increased S. feltiae infectivity to >95%, whereas it did not result in a significant increase in H. bacteriophora or S. yirgalemense infectivity.     

Characterisation of Cydia pomonella granulovirus from codling moths in a laboratory colony and in orchards of British Columbia*

Sequence analyses indicate that a granulovirus isolated from a laboratory colony of codling moth in British Columbia, Canada is identical or very similar to the Mexican isolate of Cydia pomonella granulovirus (CpGV-M). In addition to codling moth, the vims was found to cause mortality of secondary leafroller pests of North American orchards, including the fruit tree leafroller and the obliquebanded leafroller. A survey in apple growing regions of British Columbia indicates that 23% of the wild population of codling moth larvae carry CpGV.     

Fruit Bins Washed with Steinernema carpocapsae (Rhabditida: Steinernematidae) to Control Cydia pomonella (Lepidoptera: Tortricidae)

A bin washing apparatus (drencher) was used to treat late-instar codling moth, Cydia pomonella Linnaeus-infested wooden fruit bins with the entomopathogenic nematode Steinernema carpocapsae . Sentinel late-instar codling moth larvae were placed in strips of wood, made from dismantled bins, that were subsequently screwed into the corners of real bins before treatment. Pre-soaking the bins with water prior to treatment and covering the bins with a plastic tarp post-treatment, to maintain a high humidity, significantly increased sentinel codling moth mortality. Treatment for 1 min with 50 S. carpocapsae infective juveniles mL -1 water resulted in more than 80% mortality over 6 h in which the nematodes were continuously pumped through the drencher. The temperature of the water in the bin washer was ≤15°C and the chlorine in the city water was too low to harm the nematodes. The key disadvantage of washing the bins was the labour involved. It may be possible to reduce post-treatment holding time and increase codling moth mortality by holding the bins at a higher temperature post-treatment, however this parameter was not included in the study. The presence of the codling moth granulosis virus within the larvae did not influence the nematode-induced codling moth mortality over a range of concentrations and two temperatures in laboratory trials.     

Pheromone pre-exposure and mating modulate codling moth (Lepidoptera: Tortricidae) response to host plant volatiles

Abstract 1 Two codling moth Cydia pomonella kairomonal attractants, ethyl (E,Z)‐2,4‐decadienoate (pear ester) and (E)‐β‐farnesene, were tested in an insecticide‐sprayed apple orchard and an orchard treated for mating disruption with synthetic pheromone (E,E)‐8,10‐dodecadienol (codlemone). Male captures with pear ester were higher in the pheromone‐treated than in the insecticide‐treated orchard, whereas captures with (E)‐β‐farnesene were not different. Subsequent wind tunnel experiments confirmed that pre‐exposure to sex pheromone codlemone increased the behavioural response of codling moth males to pear ester. This supports the idea that male attraction to the plant volatile pear ester and sex pheromone codlemone is mediated through the same sensory channels. 2 Pear ester is a bisexual codling moth attractant and even captures of female moths were significantly increased in the pheromone‐treated orchard. In the laboratory wind tunnel, pheromone pre‐exposure had no effect on female response to pear ester, but significantly more mated than unmated codling moth females flew upwind towards a pear ester source. Differences in mating status in insecticide‐treated vs. pheromone‐treated orchards may thus account for the differences in female trap captures with pear ester. 3 These findings are important with respect to monitoring of codling moth with pear ester in mating disruption orchards. They also emphasize the importance of host plant volatiles in pheromone‐mediated mating disruption, which has been neglected to date.     

Compatibility of codling moths Cydia pomonella (Linnaeus) (Lepidoptera: Tortricidae) from South Africa with codling moths shipped from Canada

The sterile insect technique (SIT) has been successfully applied against codling moth Cydia pomonella (Linnaeus) (Lepidoptera; Tortricidae) in British Columbia since 1992 where the mass‐rearing facility produces between 15 and 16 million moths per week. Due to the seasonality of this pest, the facility is only fully utilized for part of the year. The time and expense of implementing SIT against codling moth in South Africa may be substantially reduced if moths from Canada were sexually compatible with those from South Africa. In addition, because the pome fruit‐growing season in both countries is opposite, the programme in Canada might benefit by maintaining moth production year‐round and selling moths to South Africa. Semi‐field studies in small cages and release‐recapture studies were conducted in an unsprayed apple orchard in South Africa to assess mating compatibility of laboratory‐reared codling moth from Canada and wild codling moths from South Africa. The results suggest that Canadian codling moth males were equally attracted to calling Canadian and South African females despite the fact that Canadian moths had been transported (from Canada to South Africa) for 48 h as both pupae and adults. The data also suggest that at lower field temperatures Canadian moths were more active than South African moths. Results from the release‐recapture field trials indicated that Canadian and South African males were equally attracted to Canadian and South African females. These results suggest that codling moths from Canada and South Africa are fully compatible and indicate that Canadian moths can be used for SIT studies in South Africa. As these studies were conducted with moths from two very different climatic and time zones, it is proposed that populations of codling moth in other pome fruit production areas may also be compatible with the Canadian moths.     

Net enclosures disrupt codling moth dispersal not establishment

1. Apple growers have pursued the use of exclusion netting to reduce pesticide inputs and maintain control of codling moth.
2. It is uncertain if these nets provide a physical or behavioural behaviour, and if they prevent codling moth establishment.
3. To address this, we conducted field trials testing the ability of commercial netting to exclude codling moth using small (3 trees) and large (48 trees) cages. Experiments were conducted to evaluate both the permeability by releasing marked sterile moths either inside or outside the small cages, and the establishment of wild codling moth in the large cages.
4. Results from the small cage study showed that netting reduced codling moth movement out of the netted plots, but virtually prevented codling moth dispersal into the plots. Codling moth capture inside the large cage plots was significantly less than in uncaged plots, but no differences were found in codling moth damage at harvest.
5. These results indicate that the netting is physically permeable to codling moth adults; however, it acts as a behavioural barrier to moth immigration. These outcomes agree with existing literature that net enclosures can provide a non-insecticidal tool to improve codling moth management, although it may not be a stand-alone technique.

     

Releases of Trichogramma platneri (Hymenoptera: Trichogrammatidae) in Apple Orchards under a Sterile Codling Moth Release Program

Based on the premise that augmented host numbers may help multiply and support parasitoid populations, the egg parasitoid Trichogramma platneri Nargarkatti was released in apple orchards which were participating in a sterile codling moth, Cydia pomonella (L.), release program. Nonviable eggs resulting from matings involving at least one sterile codling moth partner can be successfully parasitized by T. platneri. Grain moth-reared, as well as codling moth-reared, T. platneri were released either in the spring or in the spring and summer oviposition period(s) of wild codling moth. Sentinel codling moth eggs were hung weekly, for 3-day periods, from May until September to determine fluctuations in T. platneri populations both during and between releases. Low numbers of wild or nonviable codling moth eggs or other susceptible host eggs resulted in sufficient eggs to maintain low spring- or summer-introduced T. platneri populations. T. platneri reduced codling moth damage in trees in which the Trichogramma were released.     

Diversity and distribution of entomopathogenic nematodes (Rhabditida: Steinernematidae and Heterorhabditidae) in Turkey

The diversity and distribution of entomopathogenic nematodes in thefamilies Steinernematidae and Heterorhabditidae were assessed throughout anextensive soil survey in Turkey during 1999 and 2000. Entomopathogenic nematodeswere recovered from six out of seven regions sampled, with 22 positive sites(2%) out of 1080 sites sampled. A single nematode isolate was recovered at eachof the positive sites, of which 15 were steinernematid isolates and seven wereheterorhabditid isolates representing a total of four species. Based onmorphometric and molecular data, the nematode species were identified asHeterorhabditis bacteriophora, Steinernemafeltiae, S. affine, andSteinernema n. sp. The most common species was S.feltiae, which was isolated from 10 sites in six regions, followed byH. bacteriophora from seven sites in five regions,S. affine from four sites in two regions, andSteinernema n. sp. from one site. Heterorhabditisbacteriophora and S. feltiae have been found inmany parts of the world, whereas S. affine, so far, hasonly been recovered in Europe until our survey. Steinernemaaffine was isolated from the European (Marmara) as well as theAsiatic region (Middle Anatolia) of Turkey. A new undescribedSteinernema sp. was isolated from the most eastern region(East Anatolia) of Turkey. Soils of the positive sites were classified as sandy,sandy loam, or loam (68.2%) and sandy–clay–loam or clay loam (31.8%) and the pHranged from 5.6 to 7.9. The habitats from which the entomopathogenic nematodeswere isolated were broadly classified as disturbed (59.1%), which includedagricultural fields and poplar planted for lumber and wind breaks, andundisturbed (40.9%), which included pine forest, grassland, marsh and reed sites.Steinernema feltiae, S. affine, andH. bacteriophora were recovered from both disturbed andundisturbed habitats. The new Steinernema sp. was recoveredfrom grassland. Our survey showed that these nematodes occur widely throughoutTurkey, but at a frequency below that reported for other parts of the world.