Morphology of female sex pheromone gland in the horse chestnut leafminer Cameraria ohridella (Lep., Gracillariidae)

Abstract: The internal and external morphology of the female sex pheromone gland in Cameraria ohridella Deschka & Dimic, an European pest on Aesculus hippocastanum L., has been investigated by histological and electron microscopic techniques. The gland consists of a single layer of modified epidermal cells in the dorsal part of the intersegmental membrane between the eighth and ninth abdominal segments and laterally extends to the posterior apophyses. The epithelium contains large columnar- and cone-shaped cells with basally situated nuclei. The cuticle, which covers the glandular region, has a wrapped appearance and is divided into a hyalin and thickened endocuticle and a thin outer epicuticle: it considerably expands when the gland is protruded and provides a sufficiently large surface for evaporation of the pheromone. The cuticle does not show any orifices of pore channels. In the retracted position, the gland is folded within the body cavity of the seventh and eighth abdominal segments but is exposed to the environment by extension of the abdominal tip along with female calling. In virgin females, pheromone glands are well developed at least within the first days after eclosion; if copulation occurs, glandular epithelia degenerate soon. According to the current classification, the glandular type of C. ohridella most easily is consistent with eversible dorsal scent folds that are widely distributed amongst diverse taxa of Lepidoptera. However, this is the first report on the morphology of pheromone glands in the Gracillariidae.     

Visual, semi-quantitative assessments allow accurate estimates of leafminer population densities: an example comparing image processing and visual evaluation of damage by the horse chestnut leafminer Cameraria ohridella (Lep., Gracillariidae)

Abstract: Qualitative or semi-quantitative visual assessments are most often used for estimating population size of herbivorous insects. The precision of these estimates, however, is often difficult to establish. A 'simulation game' with the horse chestnut leafminer, Cameraria ohridella Deschka & Dimic (Lep., Gracillariidae) shows that visual, semi-quantitative assessments can provide accurate information. Damaged areas of 411 horse chestnut leaves collected in 100 sites were closely related to mine numbers despite some variability in mine and leaf size ( R ² = 0.915; n  = 411; P < 0.001). On the basis of this relationship, two methods of population assessment are compared: (i) digital image processing of leaf damage and (ii) visual assessment using a damage key reflecting the relative infested area on each leaf (0, 0%; 1, 0–2%; 2, 2–5%; 3, 5–10%; 4, 10–25%; 5, 25–50%; 6, 50–75%; 7, 75–100%). Both methods used to estimate damage presented a similar, close relationship to the 'real' numbers of mines ( R ² = 0.858; n  = 777; P < 0.001 for image processing and R ² = 0.905; n  = 777; P < 0.001 for visual assessment). The potential of using visual assessments as an accurate and fast method in situ at the tree scale is discussed.     

Sequential sampling plan for Cameraria ohridella (Lepidoptera: Gracillariidae) on horse chestnut tree

A fixed precision sequential sampling plan for estimating the density of the horse chestnut, Aesculus hippocastanum L., leafminer Cameraria ohridella Deschka & Dimic (Lepidoptera: Gracillariidae) was developed. Data were collected from 2002 to 2004 in Turin, northwestern Italy, with the aim of developing a sampling strategy for estimating populations of C. ohridella mines. Taylor's power law was used as a regression model. Sampling parameters were estimated from 216 data sets, and an additional 110 independent data sets were used to validate the fixed precision sequential sampling plan with resampling software. Covariance analysis indicated that there were not significant differences in the coefficient of Taylor's power law between heights of the foliage, months, and years. Dispersion patterns of C. ohridella were determined to be aggregated. The parameters of the Taylor's power law were used to calculate minimum sample sizes and sampling stop lines for different precision levels. Considering a mean density value of five mines per leaf, an average sample number of only 49 leaves was necessary to achieve a desired precision level of 0.25. As the precision level was increased to 0.10, the average sample size increased to 303 leaves. The sequential sampling plan should provide an effective management of C. ohridella in the urban areas, minimizing sampling time and cost, and at the same should be an effective tool to reduce insecticide applications and prevent the esthetic damage.     

Predator complex of the horse chestnut leafminer Cameraria ohridella: identification and impact assessment

Abstract:  The control of Cameraria ohridella Deschka and Dimic (Lepidoptera, Gracillariidae) by natural enemies in Europe is poor. Thus, in the past 15 years epidemic population densities of the moth regularly caused a continuous pre-mature defoliation of horse chestnut trees, Aesculus hippocastanum L.. Whereas several studies investigated the parasitoid complex of the leafminer and revealed its inefficiency, only little is known about the predators of C. ohridella . The aim of this study was to identify the predator complex in European horse chestnut stands by visual observations and exposure experiments and to assess the impact of different predator guilds on C. ohridella populations by an exclusion experiment. Of all potential predators observed, only blue tits, Parus caeruleus L., great tits, Parus major L., marsh tits, Parus palustris L. and the southern oak bushcricket Meconema meridionale (Costa) were found to prey on the pre-imaginal leafminer stages. Bushcrickets have to our knowledge never before been recorded preying on leafminers; moreover M. meridionale showed a measurable negative impact on the C. ohridella population. However, birds seem to be of greater importance as predators of C. ohridella than arthropods. Predation rates of birds ranged from 2 to 4% of leafminer populations, which is of the same order of magnitude as published parasitism rates. We conclude that the augmentation of tit densities, for example via nesting boxes in chestnut trees, should be considered as a component of an integrated control strategy against the horse chestnut leafminer.     

Native parasitoids and their potential to control the invasive leafminer, Cameraria ohridella DESCH. & DIM. (Lep.: Gracillariidae)

In spite of the fact that since the end of the eighties, the horse chestnut leafminer, Cameraria ohridella, has established itself throughout Europe, native predators such as ants and birds are not attuned to this neozoic species. In contrast, several parasitic wasp species already started to exploit the invasive horse chestnut leafminer, but until now parasitation rates are quite low, mainly because of asynchrony in the lifecycles of parasitoids and host. Only the removal of leaf litter, in which pupae hibernate, is at the moment a strategy to reduce the infestation level in the next year. Unfortunately, not only hibernating horse chestnut leafminers but also parasitoids are removed, and important resources for biocontrol are unused. In the current study, we investigated the potential efficiency of the horse chestnut leafminer parasitoid complex extracted from leaf litter in defined environments. Parasitoids were released at different densities to investigate density dependence in parasitation rates.Although seven different species were released in our experiments, only Pnigalio agraules turned out to be responsible for biocontrol of C. ohridella. We recorded parasitation rates of up to 35%. Overall, parasitation rates were independent of the leafminer density but increased fourfold if ten times more parasitoid individuals were released. Unfortunately, none of the parasitoid species could be established in the experimental units in the long run. Results are compared to other parasitoid-leafminer systems, and promotion of horse chestnut leafminer parasitoids to support natural selection and biological control of the horse chestnut leafminer is discussed.     

Pathogenicity of entomopathogenic fungi on hibernating pupae of Cameraria ohridella Deschka & Dimic 1986 (Lepidoptera, Gracillariidae). Part 1: Pathogenicity against the naked pupa

Strains from Paecilomyces fumosoroseus, Lecanicillium muscarium, Metarhizium anisopliae and Beauveria bassiana were examined in standardized Biotest to control the horse-chestnut leaf miner (Cameraria ohridella) in her pupal stage in winter. The fungi were pathogenic against the hibernating pupae of Cameraria ohridella at dose of 1.9 x 10(7) conidia/ml. They were aggressive, led to infection, death and mouldiness of naked pupae. Even at low temperature of 5 degrees C and 12 degrees C. L. muscarium strain V24 showed the highest pathogenicity after 4 weeks against this host, close followed by P. fumosoroseus strain P6. M. anisopliae strain 72 and 8. bassiana strain B412 were also pathogen but after a long-time period. Experiments gave information for general susceptibility of naked pupae of C. ohridella under low temperatures against entomopathogenic fungi. In further examinations it has to be tested, whether fungi can infected, when the pupae stay in their natural surroundings, the pupal cell in the leaf.     

Evaluation of the community of native eulophid parasitoids on Cameraria ohridella Deschka and Dimic in urban areas

The parasitoid complex associated with the exotic leafminer Cameraria ohridella Deschka and Dimic (Lepidoptera: Gracillariidae), which attacks horse chestnut (Aesculus hippocastanum L.), was studied in the urban environment of Turin (northern Italy). The studies were carried out over 5 yr after the first detection of the pest in our region in 1999. To evaluate parasitism, 438,029 leaf mines were examined over the 5-yr period, of which 29,033 were found to be parasitized (6.6%). Also, ornamental broadleaf trees attacked by other native gracillariid leafminers and located in the proximity of the target horse chestnut trees were sampled. A total of 11 parasitoid species (Hymenoptera: Eulophidae) were recorded on C. ohridella, and the most common species were Minotetrastichus frontalis (Nees), Closterocerus trifasciatus Westwood, and Pnigalio agraules (Walker). The first species accounted for >77.5% of all parasitoids collected. Cirrospilus talitzkii Boucek was found for the first time in 2005. The high population level of the pest and the low parasitism rate show that the parasitoid complex is currently inadequate to contain C. ohridella populations effectively. The most frequent parasitoids of the moth were also found on the most common broadleaf trees in the studied area, showing how native leafminer parasitoid species are able to switch to other hosts. These results show that both native and broadleaf plants species may potentially provide an important reservoir of parasitic wasps to help protect a simple biotope, such as the urban environment, from pests.     

Possibilities to control the horse chestnut leaf miner (Cameraria ohridella) in urban environments

The continuous epidemic infestation of horse chestnuts by C. ohridella is a serious aesthetic and economic problem in urban environments. The aim of the presented studies was therefore to develop control measures which meet the special needs of urban plant protection. Removal of infested leaves in autumn and appropriate disposal of the leaflitter is the only practicable measure to reduce infestation levels so far. Another two methods, which are currently in an experimental stage, had an impact on the leafminer populations. A gelformulated combination of the C. ohridella-pheromone with a contact insecticide as attract-and-kill-system proved to be efficient at low population levels. Easily applicable and safe banks of systemic insecticides turned out to be effective in killing preimaginal stages of the leafminer on young trees. The complex of parasitoids attacking C. ohridella is comparable to that of other leafminers. Nevertheless, parasitism rates are very low at present and far away from having any controlling effect on the pest insect.     

Genetic variation of horse chestnut and red horse chestnut and trees susceptibility to Erysiphe flexuosa and Cameraria ohridella

Trees of Aesculus sp. are often present in European landscapes. Common horse chestnut and red horse chestnut are seriously damaged by the fungus Erysiphe flexuosa as well as the pest Cameraria ohridella. Both pathogen and pest have spread throughout Europe. A genetic background of trees resistance still remains unknown. The present study was undertaken to estimate genetic variation of A. hippocastanum and A.×carnea grown in urban greens and their susceptibility to powdery mildew and horse chestnut leafminer. According to obtained results both species were infected by E. flexuosa in a similar degree but there were significant differences in susceptibility of particular trees (ANOVA). C. ohridella damaged only A. hippocastanum. The correlation between age of trees and degree of infestation by pathogen and pest was not observed. The significant genetic variability between two Aesculus species was revealed by the analysis of molecular variance (AMOVA) where both the intra- and the inter-species variation were found to be significant. It was shown that 73.0% of the genetic variance was contributed by the differentiation between A. hippocastanum and A.×carnea, whereas 27.0% was partitioned within species. The medium level of genetic diversity of Aesculus spp. was determined using SRAP and RAPD analyses. The mean value of genetic similarity was equal to 0.55 for common horse chestnut and 0.98 for red horse chestnut. Among 40 polymorphic SRAP and RAPD markers 17 were associated with degree of leaf damage caused by C. ohridella.     

Monitoring and dispersal of the invading Gracillariidae Cameraria ohridella

The horse‐chestnut leafminer Cameraria ohridella is an invasive lepidoptera (Gracillariidae) which has spread throughout Europe over the last two decades. The species can be detected easily because of its noticeable damage to horse‐chestnut trees leaves, which allows the easy collection of large‐scale monitoring data. It is therefore an interesting model species to study invasion processes, and this review summarizes recent progresses in that regard. The date reported here result from monitoring of C. ohridella population based on pheromone traps and visual observation of damage carried out in Belgium, France, Germany, Switzerland and the UK. The patterns of spread measured using these different monitoring method, and the key factors associated with short‐distance dispersal (within cities), and large‐scale dispersal (across landscapes) are presented and discussed.     

Digestive enzymes activity in larvae of Cameraria ohridella (Lepidoptera: Gracillariidae)

This article presents the activity of carbohydratases and proteases in the midgut of Cameraria ohridella larvae--an oligophagous pest whose preferred feeding is horse chestnuts leaves. Optimal media pH of the assayed enzymes were similar to those of other Lepidopterans. Relatively high amylase activity, as well as maltase and sucrase activities, indicates that starch and sucrose are the main digested saccharides. Trehalase activity was similar to that described in other Lepidopterans. Activities of glycosidases were significantly lower than those of disaccharidases what suggests that neither cellulose nor glycosides are important for C. ohridella. Trypsin is the main endoprotease of this pest. Like in other leaf-eaters carboxypeptidase activity was higher than that of aminopeptidase. The activity of the majority of examined enzymes increased in the following successive pest generations, which could be explained by the decreased nutritional value of older leaves. Probably this phenomenon in hydrolases activity in Cameraria is a nonspecific mechanism present at this stage of co-evolution of the horse chestnut and its pest.     

Pathogenicity of entomopathogenic fungi on hibernating pupae of Cameraria ohridella Deschka & Dimic 1986 (Lepidoptera, Gracillariidae). Part 2: Efficacy of entomopathogenic fungi against pupa in her pupal cell

The naked pupae of chestnut leafminer Cameraria ohridella are susceptible for entomopathogenic fungi Paecilomyces fumosoroseus strain P6; Lecanicillium muscarium strain V24; Metarhizium anisopliae strain M72 and Beauveria bassiana strain B412. Their activity at low temperature is advantageous especially for the use in the winter season. (Richter et al., 2007a). Therefore the possibility of infection should be examined in biotest for pupae in their pupal cells on leaf of chestnut at conditions Like in winter. Efficacy of entomopathogenic fungi is decreased, if pupae are in their pupal cells. But fungi, especially L. muscarium, proved the ability to infect pupae of C. ohridella in their pupal cells. Infected pupae moulded in their pupal cells after application. Infection doesn't come from spores. In semi field trials with L. muscarium 60% of pupae were infected in their pupal cells and died after incubation with mouldiness.     

Sex pheromone of the spotted tentiform leafminer moth Phyllonorycter blancardella (Fabr.) (Lep., Gracillariidae)

Dodecyl acetate (12:OAc) ( E )-10-dodecenyl acetate ( E 10-12:OAc) and ( E )-10-dodecenol ( E 10-12:OH) in the ratio 8 : 80 : 12 were collected by solid phase micro-extraction of the volatiles emitted by virgin signalling females of the spotted tentiform leafminer moth, Phyllonorycter blancardella . The same compounds in the ratio 8 : 79 : 13 were extracted from the sex pheromone glands of virgin signalling females of the same species. The chemical structures of the compounds were identified by gas chromatography and mass spectrometry. Trapping results obtained from field tests using synthetic components of the sex pheromone demonstrated that only one component, E 10–12:OAc, was essential for the attraction of conspecific males and should be considered as a sex pheromone.     

Within and between seasonal changes of detoxifying capabilities of Cameraria ohridella (Lepidoptera : Gracillariidae) larvae

Cameraria ohridella is an invasive leaf miner, a severe pest of horse chestnut trees. Chemical control needs recognition of intrinsic metabolic capability to cope with external stressing factors. Our tasks were to check annual effects of generation, and host tree age on detoxifying abilities of the last larval stage. Activities of CAT, SOD, GSTPX, GST CarE and AChE were assayed in the midgut of larvae from two localities during 3 years and three generations, annually. Activities of GSTPX and GST were high, but CAT and SOD were low in relation to other lepidopteran larvae. In general, the second generation larvae had the highest activity, indicating an effective defence against allelochemicals produced by the host. Effects of host tree age were significant for SOD, GSTPX and CarE activities. Significant annual differences on enzyme profiles of CAT, SOD, CarE and AChE reflected differences of temperature and precipitation between consecutive years.     

Indigenous parasitoids (Hym., Chalcidoidea) of Phyllocnistis citrella Stainton (Lep., Gracillariidae) in Jaguariúna, São Paulo State, Brazil: preliminary results

Abstract: The citrus leafminer, Phyllocnistis citrella Stainton, is native to Asia and was detected for the first time in Brazil in 1996. The objectives of this study were to conduct a survey of parasitoids attacking citrus leafminer in Jaguariúna, one of the citrus production regions of São Paulo State, Brazil, and to determine whether indigenous species were providing effective control of this pest species. The collections of new leaves were made weekly at citrus groves in Jaguariúna from October 1996 to October 1997. Of the six indigenous parasitoids which were found, Galeopsomyia fausta LaSalle was the most abundant parasitoid of P. citrella in this region, accounting for more than 90% of the specimens collected. Thus, this species is a serious candidate for biological control of the citrus leafminer in this region of Brazil. Other species collected were Horismenus sp., Cirrospilus 'sp. C', Elasmus sp., Eupelmus sp. and Conura (Ceratosmicra) sp. The mean percentage of parasitism was 39.28% (6.19−86.21%) in unsprayed orchards and 21.38% (1.33−56.63%) in orchards that were occasionally treated with fungicides, acaricides and insecticides Temik and Dipterex.     

Mitochondrial and microsatellite DNA markers reveal a Balkan origin for the highly invasive horse-chestnut leaf miner Cameraria ohridella (Lepidoptera, Gracillariidae)

Biological invasions usually start with a small number of founder individuals. These founders are likely to represent a small fraction of the total genetic diversity found in the source population. Our study set out to trace genetically the geographical origin of the horse-chestnut leafminer, Cameraria ohridella , an invasive microlepidopteran whose area of origin is still unkown. Since its discovery in Macedonia 25 years ago, this insect has experienced an explosive westward range expansion, progressively colonizing all of Central and Western Europe. We used cytochrome oxidase I sequences (DNA barcode fragment) and a set of six polymorphic microsatellites to assess the genetic variability of C. ohridella populations, and to test the hypothesis that C. ohridella derives from the southern Balkans (Albania, Macedonia and Greece). Analysis of mtDNA of 486 individuals from 88 localities allowed us to identify 25 geographically structured haplotypes. In addition, 480 individuals from 16 populations from Europe and the southern Balkans were genotyped for 6 polymorphic microsatellite loci. High haplotype diversity and low measures of nucleotide diversities including a significantly negative Tajima's D indicate that C. ohridella has experienced rapid population expansion during its dispersal across Europe. Both mtDNA and microsatellites show a reduction in genetic diversity of C. ohridella populations sampled from artificial habitats (e.g. planted trees in public parks, gardens, along roads in urban or sub-urban areas) across Europe compared with C. ohridella sampled in natural stands of horse-chestnuts in the southern Balkans. These findings suggest that European populations of C. ohridella may indeed derive from the southern Balkans.     

The effects of host plant phenology on the demography and population dynamics of the leaf-mining moth, Cameraria hamadryadella (Lepidoptera: Gracillariidae)

Abstract.

• 1 We examined the effects of variation in the timing of spring leaf production and autumn leaf fall on the survival, mortality and abundance of Cameraria hamadryadella on Quercus alba and Q.macrocarpa.
• 2 We monitored and manipulated the timing of foliation on field and potted Q.alba trees and observed the abundance of C.hamadryadella on those trees. We also monitored and manipulated the timing of leaf fall on Q.alba and Q.macrocarpa trees in the field and observed its effects on survival, mortality and abundance of C.hamadryadella.
• 3 Variation in the timing of spring leaf production has no effect on C. hamudryadella abundance. However, a warm winter and spring in 1991 led to accelerated development and the imposition of a facultative third generation in one out of ten years of observation.
• 4 In 1989, leaves fell relatively early and leaf fall in the autumn accounted for more than 50% of the mortality of C.hamudryadella. in 1990 and 1991 leaves fell relatively late and leaf fall induced mortality was substantially reduced and overwinter survival was markedly increased.
• 5 The abundance of C.hamadryadella remained constant in the spring and summer of 1990 following the previous autumn's relatively early leaf fall, but increased by 10-fold in the spring of 1991 following the relatively late leaf fall of autumn 1990. The abundance of C.hamadryadella also increased 4-fold between the summer of 1991 and the spring of 1992 after another autumn of relatively late leaf fall. We attribute these increases in abundance in part to reduced mortality because of later leaf fall.
• 6 Variation in the timing of autumn leaf fall may be responsible for initiating outbreaks of C.hamadryadella.

     

Towards understanding Cameraria ohridella (Lepidoptera: Gracillariidae) development: effects of microhabitat variability in naturally growing horse-chestnut tree canopy

International Journal of Biometeorology - Dwelling intensity of horse-chestnut miner (Cameraria ohridella) larvae in various leaves insolation and temperature was measured to determine whether this...