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IOBC-WPRS Bulletin Vol. 66, 2011


IOBC-WPRS Bulletin Vol. 66, 2011

Working Group "Insect Pathogens and Entomopathogenic Nematodes".
Preceedings of the 13th European Meeting "Biological Control in IPM Systems" at Innsbruck (Austria), 19 - 23 June, 2011.
Editors: Ralf-Udo Ehlers, Neil Crickmore, Juerg Enkerli, Itamar Glazer, Martin Kirchmair, Miguel Lopez-Ferber, Sigrid
Neuhauser, Hermann Strasser, Cezary Tkaczuk, Michael Traugott.
ISBN 978-92-9067-241-8 [XXV + 550 pp.]


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Position and perspectives of biological control in the European plant protection package
Johannes Jehle

Abstract: Driven by concerns on the human and animal health and on the environment of the use
of pesticides in the European Union (EU), the European Commission has launched a new
legislative framework, the so-called EU plant protection package, which will replace the hitherto
existing registration regulations of active substances and plant protection products. It consists of
four parts: (1) The Regulation (EC) No 1107/2009 concerning the placing of plant protection
products on the market and replacing the Directive 91/414/EEC; (2) the Directive 2009/128/EC
aiming to achieve the sustainable use of pesticides; (3) the Directive 2009/127/EC regarding to
machinery for pesticide application, and (4) the Regulation (EC) No 1185/2009 concerning
statistics on pesticide marketing and use. Whereas the Directives need to be converted into national
law in the EU member states, the Regulations are legally in force in all EU member states. This
new framework aims to harmonize the plant protection legislation in Europe and to develop
measures to reduce the risks posed from the use of plant protection products. As most biological
control agents (BCAs) underlie the same legislative procedures as chemical plant protection
products, these new regulations will also apply for most BCAs. Thus, they will have a strong
impact on future registration and use of BCAs.


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Biological control of arthropod pests in outdoor crops -present status
and future challenges

Lene Sigsgaard

Abstract: There is little use of biological control in outdoor crops. While the use of biological
control against insect and mite pests in greenhouses in Denmark had reached a high level already
ten years ago and has managed to stay high, the use in outdoor crops remains low. The total
amount of money spent on biological control in Denmark has reduced, but this is due to falling
prices, since the amount of product used has not changed or increased slightly. However, still the
use in outdoor crops is small and is to a high degree restricted to the use of Bacillus thuringiensis
against lepidopterans in high value vegetable and fruits, and to the use of predatory mites in
strawberries and predatory bugs in pear. The present paper discusses some main drivers and
barriers decisive for the share of biological control in outdoor crops, using Denmark as an example.


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Exploiting synergies to optimise the impact of entomopathogenic fungi
Tariq M. Butt

Abstract: The efficacy of entomopathogenic fungi (EPF) such as Metarhizium anisopliae and
Beauveria bassiana is greatly improved when used with sub-lethal doses of insecticide or
entomopathogenic nematodes (EPN) through additive or synergistic interactions. The latter
allows each agent to be used at reduced application rates with corresponding savings for growers.
The exact mechanism for the synergism is unclear. It is postulated that sub-lethal rates of
insecticide stress the pest making it more susceptible to infection. Some insecticides may increase
pest mobility and increase acquisition of EPF conidia and, since mortality is dose-related, this
increases pest mortality. Combined application of EPF and EPN is ergonomic and can result in
synergism but some workers have noted stronger synergy if the EPN are applied 1-3 weeks after
the EPF. There is a need to identify and optimise synergies as this strategy offers many benefits
to growers including potential savings as application rates are significantly reduced. Furthermore,
this strategy is in accord with the EU Directive 2009/128/EC.


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The evolutionary ecology of Bacillus thuringiensis; can social interactions
maintain virulence and counteract strain attenuation?

Ben Raymond

Abstract: Social evolution theory makes a number of predictions about how bacterial virulence
evolves in different environments and can have important implications for how we maintain or
improve the virulence of biopestidical bacteria such as B. thuringiensis. Investment in virulence
should not favoured in homogeneous artificial media, since social “cheats” that fail to synthesize
virulence factors should be at growth advantage. There is some evidence that biopesticide derived
strains are attenuated with respect to wild-type relatives. Selection of rifampicin resistant mutants of
B. t. kurstaki in the diamondback moth Plutella xylostella led to increases in virulence and also
reduced growth rates in artificial media, as predicted by theory.


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Systematics and taxonomy of insect pathogenic fungi: current status, future directions
Stephen A. Rehner

Abstract only


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The use of entomopathogenic nematodes in the US and issues related to genetic degradation
David Shapiro-Ilan

Abstract: Research and commercial application of entomopathogenic nematodes in North America
has a long history. In the pursuit of commercial viability, there have been a number of success
stories, but also quite a number of dead ends. In this presentation, we provide insight into new
opportunities for entomopathogenic nematodes that are on the horizon. Some novel targets that are
currently being researched include a variety of wood boring insects, weevil pests, as well as pests of
pollinators. Novel application methods and formulations that are being developed will also facilitate
expanded use of nematodes as microbial control agents. Advances in biocontrol using
entomopathogenic nematodes will also be enhanced through fundamental studies on the microbial
ecology, epizootiology, trait deterioration, population dynamics as well as basic studies on hostpathogen


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Nucleopolyhedrovirus covert infection in Spodoptera exigua: sublethal disease
and vertically transmitted infections likely provide unique opportunities
for virus-based control on greenhouse crops

Oihana Cabodevilla, Elisabeth A. Herniou, Primitivo Caballero, Trevor Williams

Abstract: The multiple nucleopolyhedrovirus of Spodoptera exigua (SeMNPV) forms the basis for
a number of highly effective biological insecticides. To assess the prevalence and diversity of
SeMNPV genotypes in natural S. exigua populations, field populations were subjected to molecular
analyses. The genotypes associated with covert infections, that are likely to be vertically
transmitted, were compared to those associated with horizontally transmitted infections. Insights
into the relationship between genetic diversity and transmission strategy were obtained by
comparing the genetic composition and insecticidal properties, of genotypes associated with
vertically and horizontally transmitted infections. The results of these studies have clear
applications in the development of custom designed biopesticidal products without the need for
recombinant DNA technologies.


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Bacillus thuringiensis sv kurstaki strain EG 2348: effect of formulation on efficacy
against tomato leafminer (Tuta absoluta)

Edith Ladurner, Massimo Benuzzi, Sergio Franceschini

Abstract: The tomato leafminer, Tuta absoluta (Meyrick), is one of the most devastating pests on
tomato. Severe losses due to this pest may occur on both fresh market and processing tomato.
Current management strategies mainly rely on the use of chemical insecticides, but their integration
with other control methods, such as microbial control agents, is of sound importance because of
resistance management, residue, toxicological and ecotoxicological issues. Bacillus thuringiensis sv
kurstaki (Btk) strains proved to be effective tools to be included into control strategies of the tomato
leafminer. In the field studies, carried out in Southern Italy, the efficacy of two formulations of Btk
strain EG2348, respectively a wettable powder (f.p. Lepinox Plus®) and a suspension concentrate
(f.p. Rapax®), was investigated. The suspension concentrate proved to be more effective than the
wettable powder.


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Understanding the toxicity of Bacillus pumilus 15.1 toward the Mediterranean fruit fly
(Ceratitis capitata)

C. Alfonso Molina, Juan F. Caña Roca, Diana García, Tania Dominguez, Antonio Osuna, and Susana Vilchez

Abstract: Bacillus pumilus 15.1 has been recently reported as a strain active against larvae of the
Mediterranean fruit fly, Ceratitis capitata (Molina et al., 2010). The 15.1 strain only reveals toxicity
against this insect when cultures are incubated at low temperature for at least 96 h before the
bioassay takes place, an interesting phenomenon worth to study and characterise it in detail. In this
study we report as well that the virulence factor is of a proteinous nature. We characterised the
protein expression profile of strain 15.1 along with its growth curve and found that the strain
overproduces insoluble proteins that are secreted during sporulation.


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Potential of a Brevibacillus laterosporus and azadirachtin combination in
immature house fly integrated management

Luca Ruiu, Alberto Satta, Ignazio Floris

Abstract: Recent results of laboratory bioassays and comparative field treatments with
Brevibacillus laterosporus (Bl) and azadirachtin [AZ] are discussed in order to evaluate their use in
combination or integration, to suppress house fly immatures in natural breeding sites of animal
farms. As determined by laboratory bioassays, the lethal effects of Bl and [AZ] are concentration
dependent and the estimated median lethal concentrations (LC50s) for second instar larvae are
1.7x108 spore/g and 24.5μg/g of diet, respectively. Applications on the manure-covered cow pen in
dairy farms, at a dosage of 3l/m2, and concentrations corresponding to LC50 levels caused a
significant fly development depression in areas treated with [AZ] (63%) and Bl (46%), compared to
the control. Preliminary laboratory observations show compatibility and interactive effects of Bl
and [AZ]. Therefore, the integration of these generally regarded as safe natural insecticides is


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Bacillus thuringiensis treatment alters larval growth, hemocytes and modulation
of Hsp70 in Rhynchophorus ferrugineus

Barbara Manachini, Mirella Vazzana, Monica Celi, Vincenzo Arizza

Abstract: To study the pathogen-host relationship, we used the model of the entomopathogenic
bacterium Bacillus thuringiensis (Bt) and Rhynchophorus ferrugineus, a quarantine pest that attacks
palm trees. In particular, we focused on the Bt stress-induced infections. We studied the effect of Bt
on larval growth, on hemocytes and on the expression of the heat shock proteins. HSPs are rapidly
synthesized in the cell after exposure to stress including pathogens. Hsp70 was evaluated in the
supernatant of the hemocyte lysate (HLS) obtained from larvae fed with Bt. This is the first time
that the presence of Hsp70 has been recorded in R. ferrugineus. Bt has negative effects on larval
growth, on total hemocytes counts and on the hemocyte type. In addition Hsp70 expression was
modulated in time (3h, 6h, 12h, 24h) in response to Bt ingestion, highlighting that Bt is a stress
factor for the R. ferrugineus. Further investigation is needed to understand the possible correlation
between the reduction of hemocytes and Hsp70 modulation.


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Preliminary selection of non-target Lepidoptera species for
ecological risk assessment of Bt canola in Sicily

Barbara Manachini, Veronica Fileccia, Silvia Ruggieri, Filippo Castiglia

Abstract: It is essential to assess the environmental risk that Bt canola resistant to Lepidopteran
pests may hold and to study its effect on species assemblages that fulfil a variety of ecosystem
functions. Environmental risk assessment can be improved through the use of an ecological
model which can be applied to a specific environment, so that local species can be classified
functionally and prioritized to identify potential test species. Several other Lepidoptera species
are also directly exposed to Bt toxin. In this paper an ecological approach was followed for
selection of non-target Lepidoptera species for ecological risk assessment of Bt canola in Sicily,
using data collected over a one-year period on Lepidoptera biodiversity on different habitats.
Non-target Lepidoptera most likely to be affected were identified and prioritized for future
testing and inclusion in risk assessments. Through use of the selection matrix, knowledge gaps
were identified for future research and guidance for the design of ecologically realistic


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Oral insecticidal activity of root colonizing P. fluorescens CHA0: a biocontrol agent
with potential to control plant pests and diseases?

Monika Maurhofer, Beat Ruffner, Maria Péchy-Tarr, Alfred Rindlisbacher, Christian Obrist, Patrik Hoegger and Christoph Keel

Abstract: The application of microbial biocontrol agents for the control of fungal plant diseases
and plant insect pests is a promising approach in the development of environmentally benign pest
management strategies. The ideal biocontrol organism would be a bacterium or a fungus with
activity against both, insect pests and fungal pathogens. Here we demonstrate the oral insecticidal
activity of the root colonizing Pseudomonas fluorescens CHA0, which is so far known for its
capacity to efficiently suppress fungal plant pathogens. Feeding assays with CHA0-sprayed leaves
showed that this strain displays oral insecticidal activity and is able to efficiently kill larvae of three
important insect pests. We further show data indicating that the Fit insect toxin produced by CHA0
and also metabolites controlled by the global regulator GacA contribute to oral insect toxicity.


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Phylogenetic position of Fit insect toxin producing pseudomonads
Beat Ruffner, Maria Péchy-Tarr, Florian Ryffel, Christoph Keel, Monika Maurhofer

Abstract: Fit produced by Pseudomonas fluorescens CHA0 is a novel insect toxin in root
colonizing pseudomonads, of which a homologue is described in Photorhabdus species.
However, occurrence and abundance of insect pathogenicity in plant-associated pseudomonads is
still unclear. An extensive screening outside the P. fluorescens complex identified strains of
Pseudomonas chlororaphis as further Fit toxin producing candidates. Sequences of five different
P. chlororaphis strains generated in this study were used to reconstruct the evolutionary history
of the Fit toxin gene and to analyse its mode of evolution. We found that P. chlororaphis is
closely associated with a small subgroup of 2,4-diacetylphloroglucinol and pyoluteorinproducing
pseudomonads, both when analyzing four housekeeping genes and the nucleotide
sequences for the Fit toxin gene. Additionally, we identified purifying selection to be the
predominant mode of Fit toxin evolution.


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Identification and characterizarion of novel vip3-like genes in
Bacillus thuringiensis strains from a Spanish collection

Leopoldo Palma, Mireya Maeztu, Iñigo Ruiz de Escudero, Delia Muñoz & Primitivo Caballero

Abstract: A total of 400 strains of Bacillus thuringiensis (Bt) have been screened to identify the
presence of vip3-like genes using a PCR-restriction fragment length polymorphism (PCR-RFLP)
strategy to detect novel vip3 genes. This allowed identification of vip3A known genes in ~80% of
the strains, whereas three new RFLP patterns revealed the presence of three novel vip3-like genes
in three different Bt strains (B5.3, B8.1, B8.3). PCR amplicons from B8.1 and B8.3 strains
produced fragments of 1646 and 1642 bp, respectively that were cloned and sequenced. These
two fragments shared homologies of ~77% with known vip3 genes suggesting they may be novel
vip3 genes. The strain B5.3 amplicon was 1620 bp and showed 91% homology with vip3Aa1
gene. The full-length sequence of this vip3A-like gene was obtained and found to be a new
vip3Ag variant and was designated vip3Ag4 by the Bt Toxin Nomenclature Committee. This gene
is 2364 bp in size, encodes a 788 amino acid (88 kDa) protein and shares between 81 and 91%
homology with known vip3A genes. Furthermore, vip3Ag4 was successfully cloned in pET-
28b(+) and expressed in Escherichia coli BL21(DE3). The protein obtained was toxic against
Chrysodeixis chalcites, Helicoverpa armigera, Spodoptera exigua, S. frugiperda and S. littoralis.


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Proteolytic processing of Bacillus thuringiensis Cry3Ca toxin
by different protease digestion treatments

María Martínez-Solís, Patricia Hernández-Martínez, Baltasar Escriche

Abstract: Cylas puncticollis (Coleoptera: Brentidae) is one of the most important biological
threats to sweet potato productivity in East Africa. The control of this insect pest is difficult due
to their cryptic feeding behaviour. Recently, the Cry3Ca toxin was found to be toxic to Cylas spp.
However, little is known about the mode of action of this Cry toxin active against C. puncticollis.
The present work studied the proteolytic activation pattern of the Cry3Ca 70kDa protoxin after
treatment with commercial enzymes, midgut juice and brush border membrane vesicles
(BBMVs). Our results showed that the activated fragment obtained after all treatments is a
polypeptide of about 50 kDa.


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In vivo selection of Cry1Aa13 toxin mutants with high affinity for intestinal proteins
of the Mediterranean fruit fly Ceratitis capitata from a phage display library

Tania Domínguez, Juan F. Caña Roca, Diana C. García, Colin Berry, Antonio Osuna and Susana Vílchez

Abstract: In this work we aim to test the hypothesis that in vitro evolution of Cry toxins and
their selection using phage display technology can be a useful strategy for developing novel
insecticides actives against non-target insects. It was demonstrated that it is possible to select Cry
toxin mutants that bind with high affinity to the intestinal proteins of Ceratitis capitata, from a
library of bacteriophages expressing Cry1Aa13 toxin mutants on their surface. Selected mutants
showed a completely different amino acid sequence in the loop2 of the domain II compared with
the lepidopteran active wild type protein. These toxins could potentially be active against this non
naturally target insect, as binding to epithelial cells proteins is the first step required for Cry toxin


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Entomopathogenic fungi as endophytes: a new plant protection strategy?
Stefan Vidal

Abstract: Since several years entomopathogenic fungi, specifically Beauveria bassiana,
Lecanicillium lecanii, and Metarhizium anisopliae, have been reported as organisms able to
colonise different plant species and plant parts, thereby maintaining their entomopathogenic
potential against various insect pests. This review discusses different aspects of the use of these
entomopathogenic fungi as a plant protection strategy. Although the endophytic growth and
establishment of these fungi can be initiated either by foliar spore application or by incorporation
into the soil, the growth and maintenance within the plant tissues need to be guaranteed during
the whole cropping season. New or improved formulation strategies need to be developed to
enhance the colonization process of these organisms in the different plant tissues. So far the
infection process of herbivores feeding on colonized plant tissues has not been studied in detail
and mycosis has not been reported in all cases. Whether production of mycotoxins by endophytic
entomopathogenic fungi may create a problem for humans consuming colonized plant parts needs
to be addressed in forthcoming studies, as well potential non-target effects on natural enemies.
The potential of endophytic entomopathogenic fungi as a new strategy for plant protection is
discussed in the light of these open questions.


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In planta detection and quantification of an endophytic Beauveria bassiana strain
Cristina López-Díaz, Blanca B. Landa del Castillo, Fº. Javier Muñoz-Ledesma, Enrique Quesada-Moraga

Abstract only


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Systemic effects of fungal endophyte seed treatment on the development
of the spotted stalk borer, Chilo partellus (Swinhoe)

Juliet Akello, Jean N. K. Maniania, Richard Sikora

Abstract: Greenhouse experiments were undertaken to investigate the systemic effects of cereal
associated fungal endophytes on the development of Chilo partellus neonates in maize. Maize
seeds were treated with seven endophytic fungal isolates and infested with C. partellus neonates,
17 days after emergence. The results indicate that seed treatment with endophytic strains
E3RF20, S4ST7, S4SU1 and G1LU3 adversely affected larval feeding and development,
resulting in less dead hearts and damage as compared to the untreated controls, four weeks after
infestation. Thus, seed treatment with the above isolates show promise for early protection of
maize seedlings. However, field studies under subsistence farming conditions in the subtropical
and tropical environments are warranted if these isolates are to be considered as an IPM tool for
lessening yield reductions attributed to stem borers.


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Effects of Metarhizium anisopliae, Bacillus thuringiensis and
new chemistry insecticide on the mortality and pupation of field populations
of Helicoverpa armigera Hübner (Lepidoptera: Noctuidae)

Waqas Wakil, M. Usman Ghazanfar

Abstract: Two dose rates of Bacillus thuringiensis (0.5, 1μg g-1), one dose rate of Metarhizium
anisopliae (1.3×106 conidia ml-1) and chlorantraniliprole (0.01ppm) each were applied alone and
in combination against second larval instar of the tomato fruitworm Helicoverpa armigera
Hübner. The overall results showed that all the treatments gave significant control of
H. armigera, however, the population from Gujranwala proved most tolerant and of Rawalpindi
was highly susceptible.


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Exploring and exploiting indigenous entomopathogenic fungi for managing the
wood boring beetle, emerald ash borer, Agrilus planipennis, in North America

George Kyei-Poku; Shajahan Johny

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Molecular diversity of the Metarhizium anisopliae lineage in an agricultural field
Bernhardt Michael Steinwender, Jürg Enkerli, Franco Widmer, Jørgen Eilenberg, Nicolai Vitt Meyling

Abstract: Entomopathogenic fungal isolates identified by morphology as Metarhizium
anisopliae may belong to different species when identified by molecular characters. We isolated
Metarhizium spp. from a Danish agricultural field using Tenebrio molitor as bait insect to assess
the molecular diversity within the soil of a single field. Isolates were analyzed using DNA
sequencing and applying SSR markers. Within the former M. anisopliae lineage, we found
M. brunneum (86.3%), M. robertsii (11.3%) and M. majus (3.4%) in the soil samples. Several
genotypes of each species were identified based on SSR markers. Differences in abundance of
the species and their genotypes suggest different adaptations to the soil environment of the
agricultural field.


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Microsatellite markers for cultivation-independent monitoring of persistence
of the entomopathogenic fungus Beauveria bassiana in soil samples

Sandhya Galidevara, Monika Bischoff-Schaefer, Jacqueline Hirsch, K. Uma Devi and Annette Reineke

Abstract: If bulk DNA extracted from soil samples should be monitored for the occurrence and
persistence of a particular strain of an entomopathogenic fungus, strain specific molecular
methods are needed. Here, we report on the applicability of five Beauveria bassiana specific
microsatellite (SSR) markers for amplification of strain specific bands in the B. bassiana isolate
ATCC 74040, which is present in the commercial product Naturalis. Discriminatory power of
these markers was initially determined on B. bassiana DNAs from a world-wide collection of this
entomopathogen. Single clearly separated peaks were evident in chromatograms after capillary
electrophoresis, which had a characteristic size for most of the respective B. bassiana isolates.
These microsatellite markers were accordingly applied for monitoring the establishment and
persistence of B. bassiana isolate ATCC 74040 in the field. Experiments were conducted in a
commercial nursery on a 100m by 25m field planted with Paeonia spp. Two defined plots within
this field each with a size of 2m x 2m were treated with a 3% Naturalis-water suspension. Soil
samples were taken before and after inoculation with B. bassiana. From the treated plots soil was
taken and analysed 4, 10 and 19 weeks, respectively, after Naturalis application. Beauveria
bassiana was successfully established and isolated from soil samples up to a period of 19 weeks
after inoculation as indicated by the amplification of B. bassiana strain specific SSR profiles
from soil DNA preparations.


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Using 454 pyrosequencing for assessing the effect of artificially applied
Beauveria bassiana on fungal diversity in agricultural soils in India

Jacqueline Hirsch, Sandhya Galidevara, Stephan Strohmeier, Martin Pfannkuchen, K. Uma Devi and Annette Reineke

Abstract: Due to optimal climatic conditions, entomopathogenic fungi (EPF) like Beauveria
bassiana are frequently used as biocontrol agents against insects in tropical countries. EPF are
usually sprayed onto the plant’s foliage or directly onto the soil. So far little is known on the
effect of applications of EPF towards the naturally occurring soil microflora. In 2010, two
agricultural fields in India were examined to define the structure and richness of the indigenous
soil fungal community and to determine the influence of artificially applied Beauveria bassiana
(strain ITCC 4688) on these populations. We performed this study by using tag-encoded 454
pyrosequencing of the fungal nuclear ribosomal internal transcribed spacer-1 (ITS-1) region. Our
study validates the effectiveness of high-throughput 454 sequencing technology for assessing soil
fungal diversity in agricultural soils and gives first insights into the effects of the application of a
microbial biocontrol agent on soil fungal community compositions.


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Investigation of Metarhizium spp. abundance in different habitat types
based on cultivation-independent detection and quantification

Salome Schneider, Katja Jacot, Franco Widmer, Jürg Enkerli

Abstract: The genus Metarhizium includes insect pathogenic fungal species, which are currently
used as biological control agents. The development of effective biological control strategies
requires detailed knowledge of their biology and the environmental factors affecting their
distribution and abundance. The goal of this study was to monitor Metarhizium spp. including
M. anisopliae, M. pinghaense, M. majus, M. brunneum, M. robertsii, and M. guizhouense in soil
using a cultivation-independent PCR-based detection and quantification tool. Soil was collected
from different habitat types including semi-natural habitats, which may provide a refuge for
Metarhizium spp. Abundance of Metarhizium spp. was significantly higher in semi-natural
habitats, i.e., improved field margins and permanent grassland compared to cereal fields.
Furthermore, uneven distribution of Metarhizium spp. was observed. Assessment of correlations
between PCR and cultivation-dependent based data demonstrated the reliability and robustness of
the PCR-based tool. The findings illustrated the value of fast and reliable detection and
quantification tools for analyzing large numbers of soil samples and for further investigations of
factors causing the uneven distribution of Metarhizium spp.


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The development of GFP-expressing Metarhizium anisopliae (Hypocreales:
Clavicipitaceae) on susceptible and resistant ticks (Acari: Ixodidae)

Dana Ment, Galina Gindin, Alice C. L. Churchill, Asael Rot, Eduard Belausov, Itamar Glazer, Michael Samish

Abstract: The fungus Metarhizium anisopliae (Hypocreales: Clavicipitaceae) is an efficient
biological control agent of various arthropods, actively penetrating host cuticles via penetration
pegs, colonizing the host body, and causing host death. Laboratory assays against various hosts
(ticks and insect) have revealed that while some hosts are highly susceptible to fungus, other
hosts are resistant to its infection. Results of previous studies of the factors involved in host
susceptibility have suggested that cuticular compounds from susceptible and resistant hosts may
differentially influence fungus development. This work describes the microscopic development
of a GFP-expressing M. anisopliae strain on susceptible tick (Acari: Ixodidae), Rhipicephalus
annulatus, and the resistant tick, Hyalomma excavatum. Conidia were observed germinating on
all hosts examined. However, the fungus was observed penetrating the cuticles of the susceptible
hosts only and never into the resistant hosts. Moreover, growth of germinating conidia and the
hyphae that developed from them was inhibited on resistant hosts and died within a few days.
Analyzing these observations provides new information on the specificity and importance of each
event of the host-pathogen interaction for a successful mycosis.


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Encapsulation materials and methods applied to biological insect control
Desirée Jakobs-Schönwandt, Rieke Lohse, Anant V. Patel

Abstract: There is still little systematic research on formulation materials and methods. Here we
will present materials, methods and applications for encapsulation of bioinsectides. The first
results of a recently granted project on mass-production and encapsulation of an endophytic
Beauveria bassiana isolate for rape plant protection will be shown.
Furthermore encapsulation of artificial CO2 sources for attract and kill capsules resulted in
beads which released CO2 under semi field conditions for 15 days and attracted western corn
rootworm larvae.


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Development of formulations based on Isaria fumosorosea (Hypocreales:
Cordycipitaceae) for the biological control of Bemisia tabaci
(Hemiptera: Aleyrodidae)

Erika Paola Grijalba, Mauricio Cruz, María V. Zuluaga, Alba Marina Cotes, Martha Isabel Gómez

Abstract: The whitefly Bemisia tabaci causes very important losses in more than 500 species of
plants such as tomato, cucumber, beans, potato, cotton, melon, peanut, soybean and others
(Rodríguez & Cardona, 2001). Two formulations (P1 and P2) designed as wettable powders
based on I. fumosorosea (Pc013) were developed for the control of Bemisia tabaci. For both
formulations, P1 and P2, two different conidia harvesting processes were evaluated. After six
months of storage at 8ºC formulation P1 germination was reduced by 34.35%, while it remained
above 85% in formulation P2. With regards to bio-controlling activity P1 showed a lower
efficacy (less than 35%), while P2 showed an efficacy above 80%. The higher efficacy and
stability of formulation P2 led to the selection of this product.


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Studies on the storage of formulated Metarhizium anisopliae conidia: effect of
temperature on conidia germination and virulence against Tenebrio larvae

Sarah Sonderegger, Verena Niedermayr, Hermann Strasser

Abstract: The entomopathogenic fungus Metarhizium anisopliae, applied under field conditions,
needs to be formulated to reduce the damage that can be caused, because of (i) UV-radiation, (ii)
high temperatures and (iii) contact with water. The aim of our work was to study the longevity
and virulence of conidia of M. anisopliae, mixed with six different anorganic fillers (Seramis®,
CN- Blähton®, Liadrain®, Clever Cat Litter®, Green Clay®, Tixosil™). The formulated and
unformulated conidia were stored over several time intervals (1 week, 1 month, 3 and 6 months)
at different temperatures (25°C, 4°C, -20°C).
No remarkable differences were seen between formulated and unformulated conidia stored
at 4°C and -20°C. Formulated conidia stored at 25°C for six months lost hundred percent of their
vitality in comparison to unformulated conidia and those, which were stored at colder
temperatures. All four expanded clay materials and Green Clay® were well suited for the storage
of a virulent M. anisopliae at 4°C and -20°C. Neither the production methods nor the different
expanded clay materials had a negative effect on the virulence of BIPESCO 5 conidia.


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Taxonomical and epizootiological studies on the arthropod-pathogenic fungi
in Poland and Austria

Cezary Tkaczuk, Stanisław Bałazy, Rudolf Wegensteiner

Abstract only


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Bacterial-type endochitinases as phylogenetic markers for
fungal entomopathogens and mycoparasites

Kakhramon Ergashev, Anaida G. Guzalova, Zafar F. Ismailov, Hans U. Schairer, Andreas Leclerque

Abstract: Two types of endochitinases referred to as bacterial or B-type and fungal or F-type are
known to operate in fungi. While F-type chitinases are generally believed to have morphogenetic
functions, B-type chitinases are often associated with pathogenesis in fungal entomopathogens
and mycoparasites. Comparative genomics has previously identified at least four fungal B-type
chitinase subfamilies termed chiB1 through chiB4. Here we explore the suitability of the chiB1
gene as a phylogenetic marker for the classification of a particular subgroup of filamentous fungi,
i.e. entomopathogenic and mycoparasitic hyphomycetes, and apply this marker to the
classification to a set of fungal isolates from Uzbekistan.


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In vitro thermal regimes of the entomopathogenic fungi
Beauveria bassiana (Vuill.) Bals. and Metarhizium anisopliae (Metch.) Sorok.

Monica Oreste, Eustachio Tarasco, Michele Poliseno, Giovanni Bubici & Oreste Triggiani

Abstract: The effect of temperature on the mycelial growth of 28 isolates of the
entomopathogenic fungus Beauveria bassiana (Vuill.) Balsam (Ascomycota, Hypocreales,
Cordycipitaceae) and five of Metarhizium anisopliae (Metch.) Sorok. (Ascomycota, Hypocreales,
Clavicipitaceae) was evaluated in vitro on Sabouraud Dextrose Agar with yeast extract (SDAY).
The five temperatures 15, 20, 25, 30 and 35°C were tested, and the fungal radial growth was
measured over 15 days at two-day intervals. Radius data were used to calculate the growth rate
(mm d-1) per each temperature, isolate and replicate by performing linear regressions, and growth
rate data were used for an analysis of covariance. Optimal and maximum temperatures and
maximum growth rate were estimated by fitting a modified β function to the growth rate data.
The thermal characterization of entomopathogenic fungal candidates for microbiological pest
control is essential to assess their applicability in the field. According to the Bio - Rational
approach, virulence against the target insect should be considered together with the adaptability
to the thermal condition in the habitat involved. In particular, tolerance to high temperatures is a
prerequisite for the application of entomopathogenic fungi in microbiological pest control in the
Mediterranean areas.


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Isolation of Beauveria bassiana from soil and its horizontal infection
among overwintering Colorado potato beetle – Leptinotarsa decemlineata
(Coleoptera, Chrysomelidae)

Medea Burjanadze

Abstract only


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The action of Beauveria bassiana introduced strains on the Colorado potato beetle
Tsisia Chkhubianishvili, Manana Kakhadze, Iatamze Malania, Levan Ninua

Abstract: The Colorado potato beetle, Leptinotarsa decemlineata, is a major pest insect of
vegetable crops in Georgia. Key elements of our present investigations are the evaluation of
introduced Beauveria bassiana strains for Colorado potato beetle management. Results of our
laboratory and field experiments demonstrated activity and a perspective for biological control.


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Prevalence of the species Beauveria pseudobassiana among tick-associated
fungal isolates from the Republic of Moldova

Polina V. Mitkovets, Natalia V. Munteanu, Galina V. Mitina, Yuri S. Tokarev, Alexandr A. Movila, Ion Toderas, Regina G. Kleespies, Andreas Leclerque

Abstract: Fungal strains isolated from ixodid ticks in the Republic of Moldova were genetically
characterized using the ribosomal RNA operon internal transcribed spacer (ITS) regions together
with the 5.8S rRNA gene as well as the nuclear genes ef1α encoding the alpha subunit of
eukaryotic translation elongation factor 1α as phylogenetic markers. On the basis of the sequence
data obtained, eight out of ten isolates were assigned to the Beauveria species B. pseudobassiana.
The two remaining isolates were consistently characterized as Isaria farinosa and as an
Aspergillus species, respectively. Further work to elucidate if the prevalence of the species B.
pseudobassiana in ticks is or not a regional phenomenon is in progress.


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Artificial hideouts with entomopathogenic fungi: A strategy for biological control
of the plum fruit moth Cydia funebrana?

Dietrich Stephan, Melanie Herker

Abstract: Especially in organic agriculture, Cydia funebrana (Plum fruit moth) is a serious pest
of plum fruits. Therefore, we investigated whether the insect can be controlled by artificial
hideouts treated with conidia of entomopathogenic fungi. Because we were not able to establish a
mass rearing of C. funebrana we did additional experiments with the related species Cydia
pomonella, Cydia molesta and Eupoecilia ambiguella. In previous experiments we proofed that
corrugated cardboard and bark mulch substrates were accepted for pupation. Additionally, we
investigated the effect of water and oil based formulation of conidia of Beauveria bassiana. The
presented data demonstrate that vegetable oils have a dramatic effect on larvae and oil-based
formulations can improve the efficacy of entomopathogenic fungi. In case of C. pomonella a
mortality of 46% and 92% was determined for Tween80 and sunflower oil, respectively. The
addition of Beauveria bassiana conidia did not enhance the mortality but even when low
concentrations of conidia (104conidia/ml) were applied in oil, 90% mycosis was achieved. In
contrast high concentrations of conidia were needed to achieve at least 70% mycosis when
formulated in water containing Tween 80. In further experiments we compared the efficacy of
B. bassiana conidia formulated in sunflower and rape oil. The results demonstrate that both oils
caused a high larval mortality and that the mycosis of the larvae was significantly higher when
conidia were formulated in rape oil. As we did not achieve high mortality with B. bassiana, we
tested other entomopathogenic fungi like Lecanicillium lecanii, Isaria fumosorosea and
Metarhizium anisopliae. M. anisopliae and I. fumosorosea caused the highest mortality to all
moth species. Furthermore, C. pomonella seems to be more sensitive to entomopathogenic fungi
than C. funebrana. In additional experiments we investigated the persistence of M. anisopliae
formulated in oil or water and two I. fumosorosea isolates formulated only in water on chipped
wood under natural conditions. The results demonstrate that the oil itself maintained its efficacy
over the observed time of two months. Furthermore, the efficacy of the three tested isolated in the
water based formulation was influenced by the weather condition. Better results were obtained
with I. fumosorosea compared to M. anisopliae.


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Efficacy of Beauveria brongniartii as Melocont in the control of the
European cockchafer (Melolontha melolontha)

Barbara H. Łabanowska, Halina Bednarek

Abstract: The European cockchafer is a dangerous soil pest of strawberry, sour cherry and apple
trees as well as other fruit orchards in some regions of Poland. The first experiments concerning
the control of white grubs of Melolontha melolontha using the fungus Beauveria brongniartii
(product Melocont) were conducted in 2009-2010 at the Research Institute of Pomology and
Floriculture in Skierniewice. The fungus B. brongniartii applied at the end of May 2009 to bare
soil (without growing plants), in young plum orchards and in a nursery on a field with Spiraea
japonica considerably reduced the number of pest insects. The best results were obtained with
Melocont on a field with white mustard (2009) when B. brongniartii was applied at the rate of
50kg/ha at the end of May and in the following year (spring 2010) at the lower rate of 30kg/ha.


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Isolation and characterisation of entomopathogenic fungi from phylloplanes
and soil of different Spanish ecosystems, and insecticidal activity
of their fungal extracts

Inmaculada Garrido-Jurado, María Fernández-Bravo, Enrique Quesada-Moraga

Abstract only


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Entomopathogenic fungal infections of hibernating pupae of horse chestnut moth
Cameraria ohridella Deschka & Dimic

Alicja Sierpinska , Katarzyna Kubiak

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Determination of mortality factors of Bemisia tabaci in cotton with
Lecanicillium lecanii applications for its control

María Victoria Zuluaga, Edgar Herney Varón, Camilo Piedrahita, Martha Isabel Gómez

Abstract: The effect of a Lecanicillium lecanii based biological control product on Bemisia
tabaci was assessed at a filed managed according to Integrated Crop Management rules (ICM)
field and farmer managed fields in the Tolima department of Colombia. An instar dependent
survival was seen for N4 nymphs in the farmer managed field, in which chemical insecticides
were used, while the ICM field showed an N4 survival of 16%. Mortality was mostly related with
the activity of L. lecanii, parasitoids and predators.


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Potential of Aphidius colemani as a vector of entomopathogenic fungi
Lecanicillium spp.

Daigo Aiuchi, Yuuna Saitou, Junya Tone, Masanori Koike

Abstract: The ability of Aphidius colemani to vector Lecanicillium spp. conidia during host
searching and/or oviposition to a colony of uninfected Aphis gossypii, and the ability of
A. colemani to discriminate Lecanicillium spp. infected aphid from uninfected individual was
demonstrated. In bioassays for discrimination ability of A. colemani against fungal infected aphid,
there were no significant difference between control plot and Lecanicillium spp. treated plots on
the number of ovipositional behaviours. Only fungus-infected aphids covered with mycelia were
avoided as host insect for oviposition. Results indicate that A. colemani does not have the ability
for discriminate between healthy and Lecanicillium spp. infected aphids for a period of up to 48h
after infection. In bioassays to assess the fungal vectoring ability of A. colemani to aphid
population, the aphid population increased ca. 26 times in control plot during 10 days. In
contrast, aphid population did not increase in plots with A. colemani release up to 8 days after
parasitoid release and then exponentially increase in 10 days. In Lecanicillium treated parasitoid
release plots, aphid density was maintained at low level up to 10 days after parasitoid release.
Considering that the number of mummies of A. gossypii was at the same level in parasitoid and
parasitoid + fungi plots, the decrease of the aphid population density might have been an effect of
Lecanicillium vectored by A. colemani. In this study, it was revealed that Lecanicillium spp. can
act additively to control the aphid by A. colemani vectoring.


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Biological control of greenhouse whitefly, Trialeurodes vaporariorum by
entomopathogenic fungus Lecanicillium muscarium hybrid strain 2aF43
in greenhouse

Hiroto Shinomiya, Daigo Aiuchi, Willem J. Ravensberg, Masayuki Tani, Masanori Koike

Abstract: The Lecanicillium muscarium hybrid strain 2aF43-formulation (average of conidial
concentration 1.7×104spore ml-1), which was obtained by protoplast fusion and shown to have a
high control potential in vitro, and the Mycotal®-formulation (3.0×106spore ml-1), which is a
commercial biological agent with a high virulence against greenhouse whiteflies on tomato plants
in greenhouses, was investigated. On untreated tomato plants, the number of adult whiteflies
significantly increased to 354 adults per leaf during 8 weeks. In 2aF43-formulation and
Mycotal®-formulation plots, adult density remained at low levels (0.6 to 15.9 adults per leaf and
0.5 to 11.3 adults per leaf, respectively) for 7 weeks. Evidence suggests that the hybrid strain
2aF43-formulation, which has a substantially lower conidial concentration than the Mycotal®-
formulation, has the potential for controlling early emergence of greenhouse whiteflies and the
possibility for long term effects in greenhouse use.


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The influence of white light exposition on the growth, sporulation and pathogenicity
of entomopathogenic fungi Isaria fumosorosea and Beauveria bassiana

Tomasz Kuźniar

Abstract: The aim of research was to evaluate the influence of white light on the growth,
sporulation and pathogenicity of entomopathogenic fungi Isaria fumosorosea and Beauveria
bassiana. The study indicated that white light was an important factor, affecting the sporulation
of entomopathogenic fungi. Fungi grown in total darkness sporulated very faintly. However the
fungi which were irradiated during the period of growth, showed considerably better ability to
produce spores. The highest production of spores was achieved when fungi were grown in total
darkness for the first three days, and next under exposure to white light. The exposure of
mycelium to white light did not influence the germination ability and pathogenicity of harvested
spores against test insects.


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Beauveria bassiana on overwintering adults of Stereonychus fraxini in Croatia
Milan Pernek, Nikola Lackovic

Abstract: The ash weevil, Stereonychus fraxini, occurred in high infestation levels in 2009
causing serious damage on different ash species in lowland forests in Lipovljani, Croatia.
Overwintering adults in moos on the bottom of trees showed high mortality rates (over 80%).
Most of these beetles were infected with the fungus Beauveria bassiana, which reduced the
population density of adults by 53.7%. The naturally occurring strain was isolated and tested on
overwintering individuals under laboratory conditions. Laboratory experiments demonstrated that
this strain was very virulent on adult weevils. Due to restrictions in use of insecticides in Croatian
forests, biological control using B. bassiana, is a promising option.


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Infecting Ips typographus (Coleoptera, Curculionidae) with Beauveria bassiana,
Metarhizium anisopliae or Isaria fumosorosea (Ascomycota)

Florian Herrmann, Rudolf Wegensteiner

Abstract: Adult Ips typographus were inoculated with Beauveria bassiana, Metarhizium
anisopliae or Isaria fumosorosea alternatively by dipping the beetles directly into conidiospore
suspensions or by forcing them to walk on spore suspension treated spruce bark. Effect of
temperature was tested by incubating experiments at 15°C, 20°C or 25°C.
Direct inoculation was successful with all three fungal species tested. Infection rates varied
between the three fungal species (75.7%-8.1%) depending on incubation temperature. Infection
rate was significantly higher only with B. bassiana at 20°C compared to the other two fungus
species. Mean life span was prolonged at 15° and shortest at 25°C with all three fungus species.
Direct inoculation caused always higher infection rates and shorter mean life span compared
to indirect inoculation with all three fungal species. Limited contact with suspension treated bark
resulted in successful infection, but duration of beetles’ contact with the conidia on bark
influenced the infection success. Infection was highest when beetles had contact with freshly
suspension treated bark for three minutes with B. bassiana and I. fumosorosea at 20°C and for
five minutes with M. anisopliae at 25°C. Infection rates were significantly lower in most cases
after shorter contact with treated bark compare to shorter contact or testing beetles after one week
incubation of log sections compared to freshly treated bark.


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Strain-specific PCR-based diagnosis for Beauveria brongniartii biocontrol strains
Ana-Cristina Fatu, Viorel Fatu, Ana-Maria Andrei, Constantin Ciornei, Daniela Lupastean, Andreas Leclerque

Abstract: A Romanian Beauveria brongniartii biocontrol isolate, strain ICDPP #1a, was
genetically compared to the producer strain of the commercially available mycoinsecticide
“Melocont®”. Analysis of 18S rRNA gene and elongation factor 1 alpha (EF1α) peptide
sequences revealed a very close phylogenetic relationship of both strains even in comparison to
further B. brongniartii strains. However, a difference in the respective 18S rRNA gene exonintron
structures was detected. Based upon this genetic difference, a PCR-based diagnosis tool
was developed that renders the two-sided positive discrimination of the strains possible.


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Morphological and genetic diversity among Polish isolates of Beauveria bassiana
Julia Budziszewska, Marta Wrzosek, Cezary Tkaczuk, Joanna Matuska-Łyżwa

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Use of mycelium overgrown corn in biological control after producing the
entomopathogenic fungus Lecanicillium muscarium on wheat

Sandra Lerche, Helga Sermann, Kathleen Schmidt, Martin Tschirner, Carmen Büttner

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Colonization of glacier foreland soils with entomopathogenic fungi
Mirjam Zoderer, Kerstin Hashold, Sigrid Neuhauser, Martin Kirchmair

Abstract: Since mid of the 19th century the Alpine glaciers are retreating continuously. The
"Rotmoos" valley glacier foreland (Tyrolean Central Alps) is 2km long with a chronosequence of
developmental stages from recently deglaciated terrain to sites which have been ice-free for 145
years at the terminal moraine dated 1858. Due to the gradual retreat of the glaciers we can now
observe all age stages of community assembly within a limited spatial scale. Such
chronosequences represent natural experiments showing the sensitivity and the resilience of
alpine ecosystems exposed to changing environmental conditions. In this study the colonization
of entomopathogenic fungi in young soils was investigated. Soil samples were taken in spring
summer and autumn at eleven sites at the glacier foreland. The abundance and density of
entomopathogenic fungi in soil were determined. Fungal isolates were identified
morphologically. Entomopathogenic fungi (Beauveria bassiana, Isaria farinosa, Hirsutella sp.,
Syngliocladium sp., the anamorph of Cordyceps militaris) as well as fungi recognized as weak
entomopathogens (Paecilomyces carneus and P. marquandii) could be detected along the
chronosequence. In the first years after deglaciation, soils are sporadically “contaminated” with
anemochorically distributed entomopathogenic fungi. In fifty year old soils entomopathogens are
frequent and diverse, similar to their possible hosts (collembolans, larvae of midges, mites etc.),
that can be found in higher abundances at older soils.


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Beauveria brongniartii in the protection of forest plantations and nurseries
against white grubs of Melolontha spp. in Poland

Alicja Sierpinska, Halina Bednarek

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Integrated control of Melolontha melolontha L. in Romanian forest nurseries
Constantin Ciornei, Ana-Maria Andrei, Daniela Lupăştean

Abstract: The research was conducted in 2008-2010 in forest nurseries in eastern Romania,
infested with larvae of Melolontha melolontha L. Biological product Melocont Pilzgerste based
on Beauveria brongniartii (Sacc) Petch has been used. It was applied in different doses (40, 50,
60kg/ha), single or in combination with sublethal doses of chemical insecticides. The treatment
effect was positive, the mortality recorded by direct action of the entomogenous fungus reached
values up to 75% and the density of larvae in the soil in the third year implementation was
considerably reduced.


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Impact of Harpellales (Kickxellomycotina, Zygomycota) on Chironomidae larvae
(Diptera, Insecta) in alpine streams

Gerald Andre, Leopold Füreder, Reinhold Pöder

Abstract: Harpellales are placed within the Kickxellomycotina, Zygomycota. Like some other
orders within the Zygomycota (e. g., Entomophthorales, Zoopagales, Asellariales), Harpellales
strongly interact with animal hosts, but it is not yet fully understood whether or not an infestation
with these fungi is beneficial or disadvantageous for their host. Literature holds evidence that the
nature of symbiosis with Harpellales depends on variables like specific host-fungus affiliations,
the physiological status of the host, and its developmental stage.
This study is focused on the impact of Harpellales on Chironomidae (Diptera, Insecta) in
alpine streams, and particularly aims at evaluating potential benefits or disadvantages of this
symbiosis in alpine streams, which represent harsh and hostile habitats.


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Genetic diversity among strains of the entomopathogenic fungus, Beauveria ssp.
as revealed by RAPD markers

A. Imoulan and A. El Meziane

Abstract: The genetic relatedness of Beauveria isolates collected from Morocco soil at different
geographic origin was determined by comparison of the products of polymerase chain reaction
amplification of genomic DNA with single arbitrary sequence oligonucleotide primers.
Beauveria isolates have a high genetic variability and they were clearly distinguished from
references isolates including B. bassiana and B. brongniartii. RAPD banding patterns did not,
however, revealed a potential relatedness with geographical origin.


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Assessment of oxalate and oosporein accumulation by endophytic
Beauveria bassiana isolates

Hermann Strasser, Pamela Vrabl, Lukas Tartarotti

Abstract: Beauveria bassiana secretes a broad range of low molecular weight metabolites, some
which are referred as relevant toxins. In submerged culture five selected endophytic B. bassiana
isolates excreted more than 20mmol l-1 oxalate but not the secondary metabolite oosporein.
Compared to the quantities of oxalate normally detected in plant tissues or in the environment it
can be concluded, that there are no risk to humans, plants and the environment.


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Potential of the strain of entomopathogenic fungus Isaria fumosorosea CCM 8367
as a biological control agent against Colorado potato beetle

Hany M. Hussein, Rostislav Zemek, Eva Prenerova

Abstract: Isaria fumosorosea (syn. Paecilomyces fumosoroseus) (WIZE) Brown & Smith
(Deuteromycota) is potentially useful for the biological control of economically important
agricultural and forest insect pests. Selection of effective, highly virulent strains is a prerequisite
for development of successful biological control agents. Our strain CCM 8367 of I. fumosorosea
was isolated from a lepidopteran pest. The aim of our study was to evaluate the effects of this
strain on larvae and pupae of the Colorado potato beetle, Leptinotarsa decemlineata (Say)
(Coleoptera: Chrysomelidae). In addition, we compared it with the effects of I. fumosorosea
strain isolated from the commercial product PreFeRal® WG (Biobest, Belgium; I. fumosorosea
strain Apopka 97 as an active ingredient). Obtained results revealed high virulence of CCM 8367
blastospores to the late last instar larvae, prepupae and pupae of L. decemlineata. Obvious
differences in virulence between our strain and the strain isolated from PreFeRal® WG were
found using fresh blastospores, where corrected mortality of CCM 8367-treated larvae eight days
after the treatment was 96.5% while mortality in Apopka 97-treated larvae reached only 54.4%.
We can conclude that the strain I. fumosorosea CCM 8367 has strong insecticidal effects on
L. decemlineata and has a potential to be implemented as a novel biocontrol agent.


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Effects of Beauveria bassiana on Frankliniella occidentalis
(Thysanoptera: Thripidae) through different routes of exposure

Andrea Boaria, Letizia Rossignolo, Alberto Pozzebon, Carlo Duso

Abstract: Beauveria bassiana is an entomopathogenic fungus widely used in biocontrol tactics
over the world. Its potential has been tested on several pest species, included the Western Flower
Thrips Frankliniella occidentalis Pergande. However, knowledge on B. bassiana-thrips interactions
is limited. In laboratory bioassays, we exposed different developmental stages of
F. occidentalis (first and second instar larvae, adults) to residual or topical applications of a
B. bassiana commercial strain (strain ATCC 74040, Naturalis®). A moderate to high mortality
was noticed for first instar larvae, late second instar larvae and adults when the two routes of
exposure were combined. The results stress on the importance to favor the contact of thrips with
B. bassiana to obtain satisfactory control of this pests. The significant reduction in thrips survival
after B. bassiana applications suggests that the latter can be included in IPM tactics.


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The influence of the larval food plants on microsporidia (Nosema mesnili P.)
infection in diapausing Pieris brassicae L. pupae

infection in diapausing Pieris brassicae L. pupae Katrin Jõgar, Luule Metspalu, Külli Hiiesaar, Angela Poomi, Marika Mänd, Irja Kivimägi, Tea Tasa and Anne Luik

Abstract: The Large White Butterfly, Pieris brassicae L., is a cosmopolitan insect, and is found
wherever cruciferous plants are grown. The P. brassicae is one of the most important pest insects
in cruciferous plants of Estonia. The experiments were carried out in the experimental laboratory
of the Estonian University of Life Sciences. In our experiment six foodplants were used: white
cabbage (Brassica oleracea L. var. capitata f. alba), german turnip (B. oleracea var. gongylodes
f. gongylodes), ornamental cabbage (B. oleracea var. acephala), broccoli (B. oleracea var.
italica), cauliflower (B. oleracea var. botrytis) and Garden Nasturtium (Tropaeolum majus L.).
The aim of the present work was to investigate the effect of larval food plants on natural infection
by mircosporidia (Nosema mesnili Paillot) in diapausing Pieris brassicae L. pupae and to assess
the influence on hibernation of P. brassicae pupae. Physiological state of P. brassicae pupae was
estimated by measuring standard metabolic rate (SMR). The infection by N. mesnili was
determined by microscopic analysis after SMR.
A statistical analysis of the results indicated that infection by N. mesnili of P. brassicae
pupae was the lowest on white cabbage. Infection was highest on german turnip and garden
nasturtium. The experiments showed that less pupae were infected by N. mesnili in ornamental
cabbage, broccoli and cauliflower variants. Healthy pupae on nasturtium plants had a higher
SMR rate than on white cabbage. Comparing infected and healthy pupae in white cabbage and
garden nasturtium variants revealed that the value of SMR was about three times higher in both
infected variants. The higher metabolic rate of infected P. brassicae pupae indicated that those
pupae would not have been able to survive winter conditions.


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Biological control of greenhouse whitefly, Trialeurodes vaporariorum by
entomopathogenic fungus Lecanicillium spp. hybrid strain in greenhouse

Daigo Aiuchi, Sayaka Horie, Toshihiro Watanabe, Satoshi Yamanaka and Masanori Koike

Abstract: The utility of Lecanicillium spp. hybrid strain 2aF43 which shown to have high control
potential at in vitro, to control greenhouse whitefly on tomato and cucumber foliage in
greenhouse were investigated. The fungal epiphytic ability on the leaf surface was
simultaneously evaluated. On tomato foliage, the number of adult whitefly in a control plot
significantly increased to 130 adults/leaf during 12 weeks. Whereas in 2aF43 and Mycotalformulation
plots, adult density was remained extremely low level (0 to 2 adults/leaf) for 10
weeks. In addition, on cucumber foliage, both fungal strains application resulted in lower density
of adults (under 2 adults/leaf) compared to control plot for 5 weeks. The density of 2aF43
propagules on both plant foliages was significantly higher than formulated Mycotal. Especially,
2aF43 was detected in high density (6.6×104 cfu/cm2) on tomato foliage, indicating possibility
that not only persisting, this strain also growing on foliage under given conditions. Evidence
suggested that hybrid strain 2aF43 has the potential for controlling early occurrence of
greenhouse whitefly and the possibility for long term effect in greenhouse use.


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Are entomopathogenic nematodes effective biological control agents
against the cereal leaf beetle (Oulema melanopus [L.])?

Žiga Laznik, Tímea Tóth, Tamás Lakatos, Matej Vidrih, Filip Vučajnk, Stanislav Trdan

Abstract: The efficacy of entomopathogenic nematodes (EPN) in controlling Oulema melanopus
on winter wheat was tested in a field experiment. A Slovenian strain of Steinernema carpocapsae
(C101) was compared with the commercial product NemasysC and the insecticide thiametoxam.
The highest yield was obtained when thiametoxam was used (7.3 ± 0.13t ha-1), while the lowest
yield was in the control treatment (5.6 ± 0.16t ha-1). In treatments with EPN we determined
statistically significant differences between both strains. Weaker response was recorded for the
commercial strain NemasysC (low nematode concentration: 5.96 ± 0.05t ha-1; high conc.: 6.1 ±
0.15t ha-1), while the Slovenian strain provided higher results (low conc.: 6.6 ± 0.14t ha-1; high
conc.: 6.8 ± 0.2t ha-1). In the field experiment different EPN concentration did not result in
statistically significant differences in wheat yield.Observing the population dynamics of the pest,
it has become apparent that EPN are effective biological agent in controlling cereal leaf beetle.


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Efficacy of entomopathogenic nematodes against the larvae and adults
of the tomato leafminer Tuta absoluta in soil treatments and
their compatibility with the insecticides used against this insect

Fernando Garcia-del-Pino, Xavier Alabern and Ana Morton

Abstract: The efficacy of soil treatments of three species of entomopathogenic nematodes (EPN)
(Steinernema carpocapsae, S. feltiae and Heterorhabditis bacteriophora) against Tuta absoluta
larvae and adults was examined under laboratory conditions. The effect of some insecticides used
against T. absoluta in the survival, infectivity and reproduction of these native species of EPNs
was also evaluated.
Soil application of EPNs gave a high control of larvae when falling into the soil for
pupation: 52.3%, 100%, and 96.7% efficacy was recorded for S. feltiae, S. carpocapsae and
H. bacteriophora, respectively. The mortality of the adults emerging from soil was 0.5% for
S. feltiae and 79.1% for S. carpocapsae. The insecticides tested have a negligible or moderate
effect on EPN survival, infectivity and reproduction. For all insecticide treatments and exposure
times, H. bacteriophora was more sensitive than S. feltiae and S. carpocapsae. Nematodes that
survived were able to infect G. mellonella larvae with no significant difference from the control.
The larvae affected by the insecticides served as suitable hosts for the infection and reproduction
of the nematodes. These results suggest that the larvae of T. absoluta falling from leaves
following insecticide application could be a suitable host for nematodes, thereby increasing their
concentration and persistence in the soil. This study demonstrates the suitability of soil
application of EPN for controlling T. absoluta.


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Attraction of Heterorhabditis sp. toward synthetic (E)–ß-caryophyllene,
a plant SOS signal emitted by maize on feeding by larvae
of Diabrotica virgifera virgifera

Samuel Anbesse, Ralf-Udo Ehlers

Abstract: Most plants when damaged by herbivore insects, synthesize and release indirect
defense mechanism by release of various chemicals that attract parasitic or predatory insects,
which are natural enemies of the herbivores. When attacked by Western corn rootworm, roots of
many maize plant varieties emit (E)-β-caryophyllene that attracts the neighboring
entomopathogenic nematodes to kill the feeding pest. Through plant genetics and biotechnology
it was possible to manipulate this volatile compound in order to increase the effectiveness of
entomopathogenic nematodes in reducing the damage of the pest. In order to further use this
strategy to improve the effectiveness of Heterorhabditis bacteriophora, we investigate the
applicability of the strategy to be used for genetic selection of enhance host finding and tested
different standard laboratory bioassays using three different sand and one agar plate assay.
Synthetic (E)-β-caryophyllene and H. megidis (the strain which showed in previous investigation,
significant attraction to caryophyllene) were used. In all the bioassays tested no significant
difference were observed between the treatment and control.


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Entomoparasitic nematodes for control of the Western corn rootworm,
Diabrotica virgifera virgifera

Christina Pilz, Udo Heimbach, Giselher Grabenweger

Abstract: Entomoparasitic nematodes of the species Heterorhabditis bacteriophora, highly
virulent against larval stages of the Western corn rootworm in laboratory studies, were tested in
field experiments in the south-eastern part of Austria. Diabrotica virgifera virgifera was first
recorded at this field site in 2002, since then high population densities developed in fields with
continuous corn. Nematodes were applied to small field plots in two different formulations,
suspended in water and as granules without water, and tested against an untreated control and
Clothianidin coated seeds in five replicates. Maize plants were artificially infested with
D. v. virgifera eggs to guarantee a homogenous pest population. Treatments were evaluated by
(a) recording the emergence of adults in cages and (b) rating the damage of maize roots.
Entomoparasitic nematodes significantly reduced the emergence of adult corn rootworms from
treated plots, regardless of formulation. They were more efficient than the treatment of maize
seeds with Clothianidin. However, nematode treatments could not prevent root damage, whereas
treatment of seeds with Clothianidin was able to keep root damage below the economic
threshold. Reasons for these contradictory results are discussed.


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Steinernema feltiae to control Western flower thrips (Frankliniella occidentialis)
in potted chrysanthemum

Andrew P. Brown, Julie Graesch, Mark Howieson

Abstract: Entomopathogenic nematodes such as a Steinernema feltiae are used in a variety of
commercial growing systems to control a number of pests as part of an integrated pest
management program. One such use is S. feltiae against Western flower thrips (Frankliniella
occidentalis, WFT). Against this pest growers are using both foliar and soil applciations to target
the different susceptible lifestages. Two field trials were carried out to investigate the efficacy of
regular foliar applciations of a commercial formulation of S. feltiae (Nemasys® F, Becker
Underwood) against WFT in potted chrysanthemum compared to a standard chemical and
alternative bicontrol Beauveria bassiana strain GHA (BotaniGard, Bioworks).


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Survey of nematodes associated with terrestrial slugs in the
Western Cape Province of South Africa

Jenna L. Ross, Elena S. Ivanova, Willem F. Sirgel, Antoinette P. Malan, Michael J. Wilsona

Abstract: A survey of slug-parasitic nematodes was conducted in the Western Cape Province of
South Africa, in order to gather new data regarding diversity and distribution. 521 slugs were
collected from 35 sample sites throughout the Western Cape. Slugs were dissected and examined
for the presence of slug-parasitic nematodes. Extracted nematodes were identified using
molecular (18S rRNA gene sequencing) and morphological techniques. Nematodes were found
parasitizing slugs at 40% of sample sites and 6% of slugs were infected with nematodes. A total
of seven species of nematode were identified in the province including; Agfa flexilis, Angiostoma
sp., Phasmarhabditis sp. SA1, Phasmarhabditis sp. SA2, Caenorhabditis elegans, panagrolaimid
sp. and Rhabditis sp. Of these species, four were slug-parasitic nematodes (A. flexilis,
Angiostoma sp., Phasmarhabditis sp. SA1 and Phasmarhabditis sp. SA2), as opposed to
nematodes that form necromenic or phoretic associations. Three new species of slug-parasitic
nematode were identified during this study (Angiostoma sp. (named as Angiostoma margaretae),
Phasmarhabditis sp. SA1 and Phasmarhabditis sp. SA2).


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Bioinsecticidal ability of virulence factors released by Steinernema carpocapsae
Nelson Simões, Duarte Toubarro, Natesan Balasubramanian, Gisela Nascimento, Mónica Martinez, Yingjun Jing, You-Jin Hao, Rafael Montiel

Abstract: Steinernema carpocapsae is a biological control agent largely used to control insects.
Laboratorial and field assays evidenced the diversity of beneficial traits of this organism namely
virulence. The analysis of transcripts and proteins in the excreted secreted products of the
parasitic nematode allowed to the identification of genes and proteins participating in invasion,
evasion and host death. Some of these genes present specific features that support its use in the
genetic improvement of the nematode, in the improvement of Bacillus thuringiensis in
combination with other insecticidal genes, and in plant transformation.


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Transcriptome analysis of desiccation and heat tolerance
of entomopathogenic nematodes

Yaari Mor, Koltai, Hinanit, Salame, Liora, Glazer, Itamar

Abstract: Under natural conditions entomopathogenic nematodes (EPNs) are exposed to extreme
environmental stresses such as desiccation and heat. Only little is known on the molecular
mechanism of stress tolerance in EPNs. Since an adaptation period is needed for acquisition of
the anhydrobiotic state and survival at high temperatures, we have focused in our research on the
adaptation period of heat and desiccation stresses, affecting nematode survival. We have
characterized the heat and desiccation tolerance of selected Steinernema species and performed a
whole transcriptome analysis. Our research is expected to provide a better understanding of the
biological pathways and molecular mechanisms of EPNs heat and desiccation stress response.
This is the first report of the next generation sequencing of EPNs.


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Development of heat and desiccation tolerance of genetically selected
hybrid strains of Heterorhabditis bacteriophora during in vivo
and in vitro propagation with and without selection pressure

Samuel Anbesse, Nanette Hope Sumaya, Verena Dörfler, Ralf-Udo Ehlers

Abstract: Repeated culturing of entomopathogenic nematodes under laboratory or industrial
scale can lead to deterioration of beneficial traits. Assessment of the development of heat and
desiccation tolerance was performed on heat and desiccation tolerant hybrid strains of
Heterorhabditis bacteriophora. The test was done in in vivo and in vitro production with or
without selection pressure for heat and desiccation. Tolerance to heat and desiccation stress was
usually lower when nematodes were propagated without selection pressure but were more
pronounced when nematodes had been propagated in vivo compared to in vitro cultivation.


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Desiccation tolerance among different isolates of the entomopathogenic nematode
Steinernema feltiae (Filipjev)

Prakijan Nimkingrat, Ralf-Udo Ehlers

Abstract: Poor storage capacity is a major constraint limiting further expansion of the use of
entomopathogenic nematodes. In order to prolong shelf life, a quiescent state of the dauer
juveniles (DJs) should be induced. This can be attained by means of desiccation of DJs. In this
study, 24 natural isolations of Steinernema feltiae were exposed to desiccation stress in non-ionic
polyethylene glycol 600. The dehydrating conditions were measured as water activity, aw-value.
Non-adapted and adapted DJs were tested separately under a series of dehydrating conditions.
The mean tolerated aw-value (MW50) ranged from 0.85 for the isolate NEP1 to 0.95 for FIN1,
ISR5 and PAL4 when not adapted to desiccation stress and from MW50 of 0.822 for CR1 to 0.98
for ISR6 when adapted to the stress conditions. CR1 tolerated the lowest desiccation stress at an
aw-value for the most tolerant 10% of the population (MW10) at 0.65 when DJs had been adapted
to stress. No significant differences were recorded between all isolates in non-adapted DJs
populations MW10 was compared. No correlation between tolerance under non-adapted and
adapted conditions were found. Most tolerant isolates will now be used for cross-breeding and
subsequent genetic selection to enhance desiccation tolerance.


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Molecular phylogeny of Heterorhabditis and Steinernema and their symbiotic bacteria.
What is true and what is wrong: impact on the evolutionary history
of these organisms?

Jean-Claude Ogier, Sylvie Pagès, Patrick Tailliez

Abstract: By associating sequences available in databases and those obtained from the
entomopathogenic nematodes preserved in our own collection at INRA Montpellier, we obtained
new insights into the phylogeny of Heterorhabditis and Steinernema and highlighted, by treetopology
reconciliation analyses, that Heterorhabditis-Photorhabdus and Steinernema-
Xenorhabdus have evolved using different events including cospeciations; sortings and host


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Beneficial trait stability in entomopathogenic nematodes
David Shapiro-Ilan

Abstract: A number of beneficial traits such as virulence, reproductive potential, and
environmental tolerance are key factors in determining an organism’s ability to produce high
levels of efficacy in biological control. Deterioration or loss of beneficial traits during laboratory
or industrial culture production is detrimental to biocontrol efficacy. During in vivo production,
both partners in the nematode-bacterium complex can undergo change, which contributes to
reduction in beneficial traits. The nematode’s bacterial symbiont also deteriorates when
repeatedly cultured in vitro. Changes in nematode beneficial traits may be due to genetic or nongenetic
(e.g., nutritional) causes. Methods to deter trait deterioration include maintenance or
improvement of media, creation of selected inbred lines, or improved cryopreservation


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A review of the use of entomopathogenic nematodes for the control of Bemisia tabaci
(Hemiptera: Aleyrodidae)

Andrew G. S. Cuthbertson, Lisa F. Blackburn, James J. Mathers, Phil Northing

No abstract


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Bacterial symbionts of entomopathogenic nematodes of the genus Steinernema
from South Italy: phenotypic features and evolutionary implications

Giancarlo Rappazzo, Carla Avanzato, Giulio Petronio Petronio, Mirella Clausi, Eustachio Tarasco, Maria Antonietta Buccheri, Pio Maria Furneri, Maria Cristina Scuderi, Maria Teresa Vinciguerra

Abstract: Three strains of Xenorhabdus kozodoii and seven of X. bovienii, obtained from
different species and isolates of the genus Steinernema (S. feltiae, S. vulcanicum, S. kraussei,
S. apuliae and S. ichnusae), were analysed by genotypic and phenotypic assays in order to
investigate in vitro biofilm production, antibiotics susceptibility and growth properties. For
identification purposes we used a genotypic analysis, such as a 16S rDNA partial sequence and
ERIC PCR profile. Phenotypic profiles were obtained by testing, among others: growth on
selective media, haemolytic activity, ß-galactosidase. Moreover, bacterial toxin was isolated and
tested in vivo on insect larvae upon direct injection into the haemolymph. The results indicate
that X. bovienii isolates present a broad phenotypic spectrum, particularly for biofilm production
and cephalosporin susceptibility, suggesting that this species underwent adaptive changes,
presumably following the evolutionary divergence of Steinernema species or isolates. X. kozodoii
diverges from X. bovienii for lower levels of biofilm production and for lactose utilisation.
However, both species synthesized a highly active toxin, which killed larvae in a manner
resembling direct infection by EPN.


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Susceptibility of larvae and adults of Leptinotarsa decemlineata to
entomopathogenic nematodes

Arne Peters, Tomke Appel

Abstract: Entomopathogenic nematodes of the species Steinernema carpocapsae,
S. feltiae and H. bacteriophora were evaluated for their efficacy in killing larvae and adults of
Colorado potato beetles. In laboratory assays on potato leaves, S. carpocapsae proved to be the
most virulent species against the larvae. On whole plant assays in a climate chamber the efficacy
was significantly enhanced by a surfactant-humectant formulation while the formulation alone
resulted in a non-significant increase in larval mortality compared to the untreated control. Adults
beetles could not be controlled by application of nematodes to the soil but were killed after being
in contact with S. carpocapsae in a gel-formulation for as little as 30 minutes.


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Nematode infections are affected by insect clotting system
Pavel Hyrsl, Pavel Dobes, Zhi Wang, Ulrich Theopold

Abstract only


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Biocontrol of leopard moth, Zeuzera pyrina L. (Lep.: Cossidae) using
entomopathogenic nematodes in Iran

Mahbobeh Ashtari, Javad Karimi, Mohammad Reza Rezapanah and Mahnaz Hassani-kakhki

Abstract: Walnuts produced in the Juglans region is among Iran's strategic products and the
Leopard moth, Zeuzera pyrina L. (Lep.: Cossidae) is the key pest of this tree. Difficulty in
chemical application against this caterpillar and special habitats of its larvae encouraged us to
address efficiency of some entomopathogenic nematodes (EPNs) against different larval stages.
Native isolates of EPNs were isolated from soil habitats of this pest in Arak, Markazi province,
Iran. Using ITS and D2/D3 expansions of 28S sequences they were identified as Heterorhabditis
bacteriophora, Steinernema feltiae and S. carpocapsae. In laboratory assays,the susceptibility of
second, third and fourth larval stages to commercial products of S. carpocapsae and
H. bacteriophora, and native strain of H. bacteriophora were studied. For field application, both
strains were used via injection of nematode suspensions into the galleries bored in tree stems or
branches. This study showed that both tested nematodes at 2,000 IJs/larva proved to be effective
on Z. pyrina. The results indicated the higher efficiency of S. carpocapsae as compared to
H. bacteriophora on larvae of this moth.


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Effect of the timing of application on efficacy of entomopathogenic nematodes
in control of Hylobius abietis (L.)

Iwona Skrzecz, Elżbieta Pezowicz, Dorota Tumialis

Abstract: The large pine weevil (Hylobius abietis) is a pest of great economic importance,
damaging young coniferous plantations. In search for new methods to reduce H. abietis
populations, the experiments were conducted aiming at the evaluation of preferred timing of
application (June or September) of entomopathogenic nematodes to decrease the number of pest
larvae developing in Scots pine (Pinus sylvestris) stumps. The treatment with Steinernema
carpocapsae and Heterorhabditis downesi at a rate 3 million specimen/stump.against H. abietis
larvae in P. sylvestris stumps applied in June indicated low efficacy. Nematode infection was
observed in approximately 19% of the large pine weevil larvae collected from the stump. Higher
efficacy of treatments at the same rate was recorded when nematodes had been applied in
September. Spraying of litter around Scots pine stumps resulted in infection of 85% of the larvae
developing in the stumps. These results were influenced by the weather conditions –
unfavourable for nematode vitality during summer treatment and favourable during autumn


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The effect of the number of bacteria Photorhabdus luminescens
(Thomas and Poinar, 1979) on the population dynamics of nematodes
Heterorhabditis megidis (Poinar, Jackson and Klein, 1987) in in vitro culture

Dorota Tumialis, Elżbieta Pezowicz, Anna Mazurkiewicz, Iwona Skrzecz, Elżbieta Popowska, Kornelia Kucharska

Abstract: The study aimed at estimating the correlation between the nematode Heterorhabditis
megidis (strain KV–136) development and the number of bacterial colonies of Photorhabdus
luminescens. Analyses concentrated on the dynamics of the population growth of H. megidis in
liquid cultures at different aeration (160rpm, 200rpm), at constant biotic and abiotic parameters
(initial dose of nematodes introduced to the culture 2,340 dauer larvae (DL)/ml, temperature
25°C, the number of bacterial colonies 0.3 x 107). Performed experiments showed that aeration at
200 rpm favourably affected the number of colonies of P. luminescens (24.14 x 107). High
density of bacteria at this aeration variant resulted in an earlier (on the fifth day of the culture)
maximum increase in the number of nematode individuals (1,239.6 H/ml) than in the culture
aerated at 160rpm.


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The effect of gold nanoparticles on the mortality and pathogenicity of
entomopathogenic nematodes from Owinema biopreparation

Kornelia Kucharska, Dorota Tumialis, Elżbieta Pezowicz, Iwona Skrzecz

Abstract: The effect of gold nanoparticles on the mortality of entomopathogenic nematodes
Steinernema feltiae from Owinema biopreparation was studied. The effect of different
concentrations of nano-Au on pathogenic properties of entomopathogenic nematodes was also
studied. It was found that mortality depended on nano-Au concentrations and on the time of
larval contact with them.


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Occurrence of pathogens and nematodes of bark beetles (Coleoptera, Scolytidae)
from coniferous forest in different region of Georgia

M. Burjanadze, M. Lortkipanidze, A. Supatashvili, O. Gorgadze

Abstract: The aim of this research was to identify pathogens and nematodes and establish their
distribution in a variety of bark beetles species from different locations and different altitude.
Various pathogen species and nematodes were observed. Gregarina typographi was present in
I. typographus (9.2-24.7%) and I. sexdentatus (35.3 %) in high prevalence; Chytridiopsis
typographi was found in I. typographus (3-6.7%) and in Tomicus piniperda (4.7%). Low
infestations with Metschnikowia sp. (1-3%) Menzbieria sp. (1.8%) were detected in
I. typographus only from one locality. Two species of parasitic nematodes was detected:
Contortylenchus diplogaster (16.9-41.9%) occurred in the population of I. typographus,
Contortylenchus pseudodiplogaster (21.2 %) in I. sexdentatus. Multiple infections were observed
with G. typographi and nematodes in combination.


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The influence of intraspecific competition and substrate on
Phasmarhabditis hermaphrodita (Nematoda: Rhabditidae)

Jiří Nermuť, Vladimír Půža, Zdeněk Mráček

Abstract: Phasmarhabditis hermaphrodita is a slug associated nematode that kills many species
of slugs and snails and is also able to live in the soil on various organic materials. This simple
laboratory study shows how intraspecific competition and substrate can influence these
nematodes. Causal relationships of lipid reserves, length, fecundity and development of
nematodes colonies to different substrates (kidney, slugs, faeces, organic matter and insect), dose
and strain (commercial and wild) were observed. We recorded some interesting trends and
behavioral patterns in this nematode. Big differences between wild and commercial strains,
especially in the fecundity and development of nematodes colonies, were also observed.


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A new entomopathogenic nematode (Steinernematidae) from Ethiopia
Tewodros Tamiru, Tesfaye Hailu, Lieven Waeyenberge, Ralf-UdoEhlers, Vladimír Půža, Zdeněk Mráček

Abstract: Three strains of Steinernema sp.n,, (Dero-1, Dero-8 and Mosisa-1) collected from
Ethiopia were characterized based on morphological, morphometric, and molecular methods.
Results confirmed that all these strains represent one undescribed species. Infective juvenile of
the Steinernema sp.n. have a body length ranging from 906-965 μm (mean 929), eight identical
ridges in lateral fields, position of excretory pore at mid of pharynx, hyaline layer occupies
approximately half of tail, ratio c´ about 3.4. First generation males without mucron whereas
second generation possesses short spine-like mucron. Slightly arcuate spicules in golden-brown
colour, with elipsoid or oblongate manubrium. First generation females without postanal swelling
and with minute protuberance on the tail tip. Second generation with postanal swelling and
protruding vulva. Based on the morphology, morphometry and DNA analysis, the new species
belongs to the glaseri-group. The closest relative species seems to be the Afro-tropical S. karii
found in Kenya. The BLAST analysis of the ITS region of the rDNA shows a similarity of 93%
with S. karii (GenBank accession number AY230173), supporting the designation to a new
species. In maximum parsimony tree the new species groups together with S. karii which is
supported by a bootstrap value of 100%.


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Possible interaction of the phoretic mite Centrouropoda almerodai on the control of
Rhynchophorus ferrugineus by entomopathogenic nematodes

Ana Morton, Fernando García-del-Pino

Abstract: The phoretic mite species Centrouropoda almerodai is associated with the red palm
weevil, using the insect to colonize new host-palms to complete its life-cycle. Entomopathogenic
nematodes are successfully used to control the red palm weevil, but the possible interaction
between C. almerodai and entomopathogenic nematodes is unknown. In this study the predation
by C. almerodai adult mites on the entomopathogenic nematode Steinernema carpocapsae was
evaluated. A low number of infective juveniles was recorded when the nematodes were exposed
to C. almerodai, confirming that the mite species feeds on S. carpocapsae.


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The abundance, distribution and natural host range of entomopathogenic
nematodes (Nematoda: Steinernematidae) in the experimental
GM maize MON 88017 field

Vladimír Půža, Oxana Habuštová, Hany M. Hussein, Zdeňka Svobodová

Abstract: The present study is a part of the project focused on the environmental impact of GM
maize MON 88017. Our aim was to assess the occurrence, abundance and distribution of
entomopathogenic nematodes and their insect hosts in the experimental field with MON 88017
GM maize and non GM cultivars. Steinernema affine was present in all maize cultivars and the
most important factor affecting it’s occurrence was probably the type of the neighbouring
environment, as EPNs were only present in plots in the part of the field neighbouring with a
forest, regardless of the maize cultivar. No effect of MON 88017 on EPN distribution was
recorded. The overall EPN density was low, probably reflecting the low number of soil insects.
Thus the biocontrol potential of such a population would be limited.


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A highly pathogenic Steinernema websteri isolated first time in Agrotis segetum and in Turkey
Gokce, C., Erbas, Z., Yilmaz, H., Demir, I. and Demirbag, Z.

Abstract only


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Histopathological changes and chitinase activity induced by Meloidogyne incognita
in resistant and susceptible potato

Mahfouz M. M. Abd-Elgawad, Marie-Claire Kerlan, Sanaa S. A. Kabeil, Farid Abd-El-Kareem, Moawad M. Mohamed, Wafaa A. El-Nagdi

Abstract only


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Susceptibility of the noxious social insect Tapinoma sessile (Say) to infection
by entomopathogenic nematodes

Nona Mikaia

Abstract: The susceptibility of the house ant, Tapinoma sessile, to entomopathogenic nematodes
(EPN) and their reproduction in this host was investigated. Mortality of ants by EPN nematodes
after 24 h exposure to S. feltiae was 21% and 28% to S. carpocapsae. After 48 h exposure the
mortality caused by S. feltiae achieved 43%, that of S. carpocapsae 52%. Mortality of insects
caused by S. feltiae after 72 h exposure was 61% and 72% for S. carpocapsae. The reproductive
potential of S. feltiae on the house ant, Tapinoma sessile, on the first, second and third day was
1500, 2000 and 2500. For S. carpocapsae it was 2800, 3300 and 1800. The reproduction of
S. feltiae on the fourth, fifth, and sixth day was much higher (3000, 3200 and 3500, accordingly)
and for S. carpocapsae – 2200, 3700 and 4000. On the seventh day the reproductive potential for
S. carpocapsae made 4200 more than S. feltiae (3800).


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Colombian granuloviruses related to Phthorimaea operculella granulovirus
isolated from the Guatemalan tuber moth, Tecia solanivora

Carlos Espinel-Correal, Xavier Léry, Laura Villamizar, Juliana Gómez, Jean Louis Zeddam, Alba Marina Cotes, and Miguel Lopez-Ferber

Abstract: The guatemalan potato tuber moth, Tecia solanivora, progressively invaded Colombia,
partially displacing the original potato tuber moth, Phthorimaea operculella. No appropriate
biological control methods exist for T. solanivora, while for P. operculella, a granulovirus,
PhopGV, is used. An extensive sampling in Colombia allowed find T. solanivora infected larvae.
Five different isolates have been characterised. All appear to be well adapted to T. solanivora,
showing control efficacies (measured by their LC50) lower than 10 OB/mm2. In addition, isolates
collected in regions where both host species coexist are also efficient on P. operculella larvae.
They are thus promising candidates for the development of biological insecticides.


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Stored potatoes in Costa Rica are efficiently protected from Phthorimaea operculella
and Tecia solanivora with an indigenous granulovirus strain

Y. Gómez-Bonilla, M. López-Ferber, P. Caballero, D. Muñoz

Abstract: The control efficiency of a Phthorimaea operculella granulovirus isolate from Costa
Rica (PhopGV-CR1) against the insect pests P. operculella and Tecia solanivora under storage
and field conditions was evaluated. The virus reduced damage by over 70% compared with the
untreated controls. These data favour the inclusion of PhopGV-CR1 formulations in IPM


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Potato crops in Costa Rica are efficiently protected from Phthorimaea operculella
and Tecia solanivora by an indigenous granulovirus strain

Y. Gómez-Bonilla, M. López-Ferber, P. Caballero, D. Muñoz

Abstract: The control efficiency of a Phthorimaea operculella granulovirus isolate from Costa
Rica (PhopGV-CR1) against the insect pests P. operculella and Tecia solanivora under field
conditions was evaluated. The virus reduced damage between 50 and 80% in fields compared
with the untreated controls. These data favor the inclusion of PhopGV-CR1 formulations in IPM


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Transcriptome analysis of the Cydia pomonella granulovirus
D. Schneider, K. E. Eberle, J. A. Jehle

Abstract: The Cydia pomonella Granulovirus (CpGV) is widely used for the control of codling
moth (C. pomonella) many apple growing regions worldwide. In recent years development of
resistance to CpGV was observed in several European countries. On the other hand, new CpGV
isolates overcoming this resistance were identified and applied in orchards with resistance. To
understand better the interaction between CpGV and its host and to determine the genetic factors
involved in the virulence of the virus the transcriptome of the virus and the expression profile of
its genome is analysed. By quantitative reverse transcription PCR (qRT-PCR) analyses, the
specific time frame of the infection process in midgut and fatbody tissue shall be discovered. The
aim of this project is to find new ways of virulence management by expanding the knowledge on
the gene regulation of CpGV in susceptible and resistant codling moth.


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The new CpGV-R5 variant overcomes the codling moth resistance to the virus
Samantha Besse, Ludovic Crabos, Olivier Soubabère, Antoine Bonhomme, Miguel Lopez-Ferber, Marie Berling, Benoît Sauphanor, Sophie-Joy Ondet

Abstract: Cydia pomonella is the most important pest in apples, pears, quinces and walnuts
orchards all over the world. Its control with chemical insecticides comes up against many
resistances developed by this insect. The biological control with a specific Granulovirus, the
CpGV, is one of the key tools for farmers, especially in organic production. After more than 20
years of generalized use of a single virus isolate, insect populations resistant to the virus have
been detected in Germany first, then in France, opening the question on the sustainability of such
control approach. Codling moth resistant populations weaken the whole production field.
By selection on a resistant insect colony, virus variants able to kill resistant insects were
obtained. The most promising is being patented. Field trials conducted in France and Germany in
2010 with one of these variants, the CpGV-R5, have shown a very interesting control level in
resistant populations, compatible with agricultural practices. The registration of CpGV-R5
variant is on-going in Europe and its equivalence with the CpGV-M isolate has been recognized
by the French authorities for its inscription at the Annex I. Its registration in France would be
obtained in 2011 under “Carpovirusine Evo2” trademark.
This example has contributed to the modification of the European regulations framework on
the registration of biological control agents, allowing the introduction of the co-evolution and of
biodiversity concepts for Baculovirus.
However, this approach requires modify the way in which the CpGV is used in orchards. A
strategy was drawn by Arysta LifeScience/NPP for the use of CpGV-R5 variant in order to
sustain the use of CpGV by farmers.


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Use of resistance overcoming CpGV isolates and CpGV resistance situation
of the codling moth in Europe seven years after the first discovery
of resistance to CpGV-M

Daniel Zingg, Markus Züger, Franz Bollhalder, Martin Andermatt

Abstract: New Cydia pomonella granulovirus (CpGV) isolates Madex Plus/Madex 100 or
Madex Max have been registered and commercialised in Austria, Germany, Holland, Italy and
Switzerland. In most plots with resistance problems the codling moth (CM) populations were
reduced to a satisfactory level by using these resistance overcoming CpGV isolates.
More highly virulent resistance overcoming CpGV isolates are available today and offer
possibilities for virulence management by changing the virus isolates every few years in order to
prevent further resistance development.


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Resistance of codling moth to Cydia pomonella granulovirus –
are there two types of resistance?

Johannes A. Jehle, Stefanie Schulze-Bopp, Eva Fritsch, Karin Undorf-Spahn

Abstract only


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Control of Oriental fruit moth and codling moth with a new Granulovirus isolate
Iris Kraaz, Markus Züger, Heiri Wandeler, Daniel Zingg, Edith Ladurner, Massimo Benuzzi, Marketa Broklova, Gyula Laszlo

Abstract: A new Granulovirus isolate for the combined control of codling moth (Cydia
pomonella) and oriental fruit moth (Grapholita molesta) has been developed and field-tested by
Andermatt Biocontrol, Switzerland. Three selected trial results from the 2010 season in Italy,
Slovakia and Switzerland are presented in this paper. The effect of the new isolate against
oriental fruit moth on stone fruit was a reduction in fruit damage of 68% (on total damage) and
77% (on active damage) and 90% (on active damage) for codling moth in apple.


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Evaluation of a microencapsulated baculovirus bioinsecticide for controlling
Spodoptera frugiperda in maize

Juliana Gómez, Judith Guevara, Laura Villamizar

Abstract: A formulation prepared by microencapsulation of nucleopolyhedrovirus SfMNPV 003
occlusion bodies (OBs) with a methacrylic acid polymer was assessed on maize plants infested
with Spodoptera frugiperda under mesh house conditions. No obvious differences in the level of
insect plant damage were observed between plants treated with formulated and unformulated
virus. On days 7 and 11, values of damages ranged from 2.5 to 17.5% for plants treated with
virus, while in control plants damages varied between 90 to 70%. Formulated and unformulated
virus at a rate dose of 1x107 OBs/ml (750g/ha) efficiently reduced insect damage under semi
controlled conditions. Biopesticide was also evaluated under field conditions in a maize crop, by
using three doses. Biological and chemical products reduced similarly the insect plant damage.
The results obtained allowed to recommend treatments at a biopesticide dose of 750g/ha
equivalent to 7.5x1011 OBs/Ha for controlling S. frugiperda under field conditions in maize crops
with applications when fresh damage reaches 15%.


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Physical maps and biological activity of seven isolates of Helicoverpa armigera
nucleopolyhedrovirus from the Iberian Peninsula

Maite Arrizubieta, Oihane Simón, Delia Muñoz, Trevor Williams and Primitivo Caballero

Abstract: In the present study, seven previously identified Helicoverpa armigera NPV
(HaSNPV) isolates from the Iberian Peninsula were subjected to molecular and biological
characterization and compared with a Chinese genotype (HaSNPV-G4). Physical maps with
EcoRI indicated that the Spanish strain HaSNPV-SP1 bore the largest genome while the others
presented deletions of varying sizes. Biological characterization revealed no differences in
pathogenicity (effective doses) among the different isolates. However, they showed variations in
virulence (speed of kill), with HaSNPV-SP1 being, by far, the fastest in killing larvae, including
the reference HaSNPV-G4. In terms of productivity (number of occlusion bodies yielded per
larva), HaSNPV-SP1 was one of the most productive isolates. In conclusion, HaSNPV-SP1
evidences highly desirable biological characteristics for its development as a bioinsecticide.


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Spodoptera frugiperda multiple nucleopolyhedrovirus as a potential biological
insecticide: genetic and phenotypic comparison of field isolates from Colombia

Gloria Barrera, Oihane Simón, Laura Villamizar, Trevor Williams and Primitivo Caballero

Abstract: Thirty eight isolates of Spodoptera frugiperda multiple nucleopolyhedrovirus
(SfMNPV), collected from infected larvae on pastures, maize and sorghum plants in three
different geographical regions of Colombia, were subjected to molecular characterization and
were compared with a previously characterized Nicaraguan isolate (SfNIC). Restriction
endonuclease analysis (REN) showed two different patterns among Colombian isolates, one
profile was particularly frequent (92%) and was named SfCOL. The physical map of SfCOL was
constructed and the genome was estimated to be 133.9 Kb, with few differences in terms of
number and position of restriction sites between the genomes of SfNIC and SfCOL. The PstI-K
and PstI-M fragments were characteristic of SfCOL. These fragments were sequenced to reveal
the presence of seven complete and two partial ORFs. This region was collinear with SfMNPV
sf20 to sf27. However, two ORFs (4 and 5) had no homologies with SfMNPV ORFs, but were
homologous with Spodoptera exigua MNPV (se21 and se22/se23) and Spodoptera litura NPV
(splt20 and splt21). Biological characterization was performed against two different colonies of
S. frugiperda, one originating from Colombia and one from Mexico. SfCOL OBs were twelve
times more potent for the Colombian colony than SfNIC OBs. SfCOL and SfNIC showed a
slower speed of kill (by ~50 h) in insects from the Colombian colony compared to the Mexican
colony, which was correlated with a higher production of OBs/larvae. SfCOL is a new strain of
SfMNPV that presents pathogenic characteristics that favor its development as the basis for a
biopesticide product in Colombia.


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Evaluation of the efficacy of two nucleopolyhedroviruses to suppress
whitemarked tussock moth populations

G. Brodersen, R. Lapointe, G. Thurston, C. Lucarotti and D. Quiring

Abstract: The nucleopolyhedroviruses of Orgyia leucostigma (OrleSNPV) and Orgyia
pseudotsugata (OpMNPV) were compared to assess their potential as microbial biological
control agents against the whitemarked tussock moth (WMTM). The influence of virus species,
dose and larval instar at inoculation on larval mortality was determined in laboratory bioassays.
Mortality rates of larvae infected with OrleSNPV and OpMNPV were comparable, although
OpMNPV had slightly higher efficacy at lower doses when fed to early-instar larvae. LT50
experiments demonstrated that OrleSNPV carried a slight but significant advantage on the time to
mortality. Manipulative field bioassays were also performed to determine the influence of preand
post-ingestion diet on the mortality caused by both viruses. Both pre- and post-virusingestion
nutrition impacted the mortality rates of larvae infected by each virus equally, which
may be associated with varying nutritional qualities, especially protein content, of the diet and
foliage used.


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Formulation effect over insecticidal activity of Phthorimaea operculella
granulovirus VG003 for controlling Tecia solanivora

Juliana Gómez, Carlos Moreno, Karen Vega, Alba Marina Cotes, Laura Villamizar

Abstract: Two formulations of a Colombian isolate of Phthorimaea operculella granulovirus
designed as emulsifiable concentrate (EC) and dispersible granules (WG) were evaluated under
laboratory, mesh house and field conditions, against the Guatemalan potato moth Tecia
solanivora. In the laboratory test both formulations showed lower potency than unformulated
virus, although there were no significant difference between their lethal concentrations. EC
formulation significantly reduced larval population, incidence and severity of damage caused by
the insect in mesh house, while WG showed no effect on any variable. Formulation type had a
different effect over insecticidal activity and EC, with a concentration of 1 x 107 OBs/ml, a
volume rate of 400l/ha and a biweekly frequency, was selected as the most promising treatment
in the field, where it produced the highest field crop protection (83%).


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Characterization and storage stability of a formulation based on a Colombian
Spodoptera frugiperda nucleopolyhedrovirus, SfNPV

Lizeth Tabima, Juliana Gómez, Laura Villamizar

Abstract: A formulation based on a Colombian isolate of Spodoptera frugiperda
nucleopolyhedrovirus, prepared by the microencapsulation of viral occlusion bodies (OBs) with a
methacrylic acid polymer was characterized to establish limits for its future quality control in
manufacture process and its stability in storage was studied. Biopesticide showed a concentration
of 109 OBs/mL, a moisture content of 1.8%, a pH of 7.1, a particle size less than 10μm, total
contaminant content lower than 105 CFU/g and a laboratory efficacy of 94%. Insecticidal activity
of formulated and unformulated virus was stable for six months of storage at 8°C and 18°C.
However, a significant reduction in efficacy was observed after six months of storage at 28°C.
Contaminants content in formulated virus remained under acceptation limit during storage at
three evaluated temperatures. Obtained results allowed to conclude that the developed SfNPV
biopesticide could be stored for six months at 8°C and 18°C without quality losses.


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Spodoptera ornithogalli nucleopolyhedrovirus: Preliminary study of Colombian isolate
Gloria Barrera, Paola Cuartas, Juliana Gómez, Judith Guevara and Laura Villamizar

Abstract: Spodoptera ornithogalli (Guenée) (Lepidoptera) is a phytophagous insect of family
Noctuidae. In Colombia, this specie had been reported in several crops including cotton and
ornamental flowers with economic importance. The Nucleopolyhedrovirus of Spodoptera
ornithogalli (SoNPV) is a natural enemy for the larvae stage. In this work, S. ornithogalli larvae
collected from citric crop were reared until dead by viral symptoms. The SoNPV obtained were
subject to morphological and biological characterization. Additionally, restriction endonuclease
analysis (REN) patterns were made using four restriction endonucleases. The size of the
polyhedra showed high variation although the most frequent range size was inferior compared to
S. frugiperda polyhedra. Mean lethal concentration (LC50) determined in neonates larvae was
1x105 OBs/mL and the yield per unit weight of the larvae was found to be 2x109 OBs/g. The
REN showed a differential pattern compared with others baculoviruses. This is a preliminary
study of SoNPV native from Colombia and could be the base for future works in order to develop
a biopesticide.


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Investigation of nucleopolyhedroviruses and experience of regulation
of forest pest populations in Latvia

Jankevica, Liga, Halimona, Julija, Metla, Zane, Seskena, Rita, Zarins, Ivars

Abstract: Baculoviruses are potential agents for the control of different forest pests. The aim of
studies was to extend the knowledge on insect viruses and to clarify their role in regulation of
pest populations. Nucleopolyhedroviruses (NPVs) were isolated from 15 pest species. Four NPV
isolates were isolated from Hymenoptera species, 11 isolates – from Lepidoptera. Isolated viruses
were used as a basis for virus preparations. NPVs isolated in Latvia were found to be active
against their corresponding pests and may be successfully used as biological control agents. The
possibilities to enhance viral infection by synergistic additives were investigated.


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Wireworm management: mitigating environmental risk with the development
of new ‘push-pull’ strategies

Robert S. Vernon and Wim Van Herk

Abstract: Many higher risk insecticides commonly used for wireworm management are now
obsolete worldwide, or soon will be, and wireworm populations and damage are on the rise
globally. The search for lower risk replacements, however, has proven to be challenging. Our
studies, for example, have shown that neonicotinoids (i.e. thiamethoxam, clothianidin and
imidacloprid) applied as seed treatments (i.e. wheat, corn, potatoes) will preserve crop stand and
yield, but this is due to wireworms entering a long-term but reversible state of intoxication rather
than mortality. We have also found that synthetic pyrethroids (i.e. bifenthrin and tefluthrin)
applied as wheat seed treatments are repellent to wireworms long enough to enable crop
establishment, but once again without wireworm mortality. In contrast, the phenyl pyrazol,
fipronil, applied at higher dosages to wheat seed and potato crops resulted in excellent crop
protection and near extermination of wireworm populations. Applications of fipronil at lower
dosages did not affect wireworm health immediately, but significant latent mortality (up to 90%)
began occurring after about 40 days. Since fipronil has a higher environmental and health risk
profile than the neonicotinoids or pyrethroids, a number of novel strategies were developed to
reduce the amount of fipronil applied per ha while maintaining crop health and significantly
reducing wireworms. A ‘blend’ of thiamethoxam (10g a.i./100kg seed) + fipronil (1g a.i./100kg
seed) on wheat seed was found to be sufficient to protect wheat stand under moderate to high
wireworm populations, as well as reducing neonate and resident populations by >90% (fipronil
rate/ha = 1.24g a.i.). Similar field efficacy was also demonstrated with ‘push-pull’ strategies
involving 1:1 mixtures of ‘repulsive’ tefluthrin-treated wheat seeds and ‘lethal’ blend-treated
wheat seeds (fipronil rate/ha = 0.62g a.i.). Push-pull strategies to merely remove wireworms from
fields, or for use as companion plantings were developed using up to 3:1 mixtures of untreated
(attractive) wheat seeds and blend-treated wheat seeds (with 3:1 seed ratios, fipronil rate/ha =
0.095g a.i.). Practical uses for these strategies are discussed.


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Crop rotation as a management tool for wireworms in potatoes
Christine Noronha

Abstract: Wireworm damage to potato tubers can result in serious economic losses for
producers. The lack of efficacious insecticides in Canada and increasing wireworm pressure has
intensified the need to evaluate other management strategies. The effectiveness of a three-year
crop rotation on tuber damage was evaluated, with buckwheat, alfalfa, brown mustard or the
standard barley/clover grown in years one and two, and potato in year three. Results showed a
significant decrease in tuber damage and an increase in marketable tuber yield following the
brown mustard and buckwheat rotations when compared to the alfalfa and barley/clover


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Trap crops as a means to manage Agriotes larvae in maize
Bettina Thalinger, Karin Staudacher, Nikolaus Schallhart, Corinna Wallinger, Anita Juen, Michael Traugott

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Seasonal patterns in the dietary choice of Agriotes wireworms
revealed by molecular analyses

Corinna Wallinger, Karin Staudacher, Nikolaus Schallhart, Evi Mitterrutzner, Eva-Maria Steiner, Bettina Thalinger, Anita Juen, Michael Traugott

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Attraction of wireworms to root-emitted volatile organic compounds of barley
Fanny Barsics, Eric Haubruge, François J. Verheggen

Abstract: The ability of wireworms (Agriotes sordidus Illiger) to orientate towards a blend of
volatiles emitted by chopped roots of barley was tested. During individual tests, the larvae chose
between the two sides of a Y-shaped olfactometer. One side was connected to a chamber
containing the chopped roots and the other was connected to an empty chamber. Wireworms
chose significantly more often the side of the olfactometer providing the blend of root volatiles.
This result underlines the importance of the identification of these compounds and their role
assessment alone or combined, as for their effect on wireworms. Such compounds could be used
in IPM strategies.


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Factors influencing monitoring of Agriotes spp. wireworms
Hilfred Huiting and Klaas van Rozen

Abstract: Wireworms tunneling into potato tubers cause quality decline to the product. Due to
the short time between initial wireworm presence in the topsoil in spring and the moment of
potato planting, decisions whether or not to control wireworms based on monitoring in spring are
insecure. Increased knowledge on temperature and soil moisture may help enhance a succesful
monitoring strategy. Two trials performed in climate chambers showed wireworms to be
increasingly active between 5°C and 25°C. The preferent soil moisture level was 20%, ranging
from 10 to 25%.


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Occurrence of Agriotes wireworms in Austrian agricultural land
Karin Staudacher, Nikolaus Schallhart, Peter Pitterl, Corinna Wallinger, Nina Brunner, Bernh ard Kromp, Marion Landl, Johann Glauninger and Michael Traugott

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How does wireworm damage in potato fields correlate with elaterid species
as well as environmental and cultivation parameters?

Nina Brunner, Patrick Hann, Claus Trska, Bernhard Kromp

Abstract: Wireworm damages are one of the major recent plant protection problems in organic
as well as in conventional potato farming. This study aimed at identifying parameters of
environment and cultivation that correlate with wireworm damages in potato as well as the
occurrence of Agriotes species. From 2007 to 2009, 29 organic potato fields, located in Eastern
Austria were investigated by wireworm-bait traps and damage ratings. Crop rotation and
cultivation methods were interrogated from the farmers for each field. Site-specific soil, climate
and landscape parameters were collected from databases. Relevant parameters were selected by
Principal Component Analysis and then correlated with the damage height and the number of
baited wireworms. The greatest differences between the research sites were caused by a regional
factor. The severity of damage seemed to increase alongside a regional gradient from the lower
warm-dry Marchfeld over central Weinviertel, both Lower Austria, towards the higher, coolhumid
and densely-wooded Wald- and Mühlviertel, Lower and Upper Austria, respectively.
Higher air temperatures in July and August seemed to diminish the damage. Single wireworm
species of the main pest-genus Agriotes showed differences in regional distribution. Only
Agriotes obscurus and Agriotes sputator correlated with the damage height.


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The importance of the identification of Agriotes larvae to implement IPM in arable crops
Lorenzo Furlan

Abstract: The implementation of IPM strategies against wireworms has been extremely difficult
because until few years ago, there was a shortage of reliable information on the key aspects of the
species concerned. In the last couple of decades information about the behavior of the larvae of
the most important species for agriculture and reliable economic threshold has been discovered.
This information can be really useful to implement IPM in arable crops if the determination of
the larva can be easily and quickly done. The results of this research with regard to the species
Agriotes brevis, A. sordidus, A. ustulatus in north-eastern Italy and the practical implications for
IPM are described. The above Agriotes species showed a different response to bait traps so it is
necessary to assess thresholds (expressed as number of larvae/bait trap) for each of the
associations crop-wireworm species provided the bait traps have been placed out in proper
conditions. Thresholds for maize crop are reported.


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Click beetles and pheromones – an overview
Miklós Tóth

Abstract: Based on experience gained from the widely used moth pheromones. It was hoped that
if pheromones of click beetles became known, they could be applied in similar ways in IPM
strategies for the control of wireworms. A brief overview is presented on results of identification
efforts of click beetle pheromones, new questions arising in the categorization of these
pheromones into the class sex or aggregation pheromones, options for the development of
female-targeted lures, and application perspectives.


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A two year click beetle monitoring with pheromone traps in Germany:
species distribution, trap specificity and activity pattern

Stefan Vidal, Torsten Block, Thilo Busch, Frank Burghause and Hans-Helmut Petersen

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Male Agriotes click beetle walking behaviour and response to sex pheromones
Carly Benefer, Rod Blackshaw, Mairi Knight and Jon Ellis

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Distribution and abundance of Agriotes lineatus L. adults on pheromone traps
in four regions in Croatia

Antonela Kozina, Maja Čačija, Renata Bažok

Abstract: The main aim of recent investigation was to determine the distribution and abundance
of Agriotes lineatus L. in four different regions of Croatia and correlate the abundance with the
prevailed climatic conditions in each region. Investigation was conducted by the mean of
pheromone traps (Csalomon) on 17 fields grouped into 7 microregions and four regions
according to the climatic data. The average capture of A. lineatus beetles/field and the average
dominance index were the highest in County of Zagreb where the most humid conditions were
present. Going to the east the amount of rainfall is decreasing and mean average capture of
beetles and the average dominance indexes became lower. In County of Vukovar-Sirmium where
the highest temperatures and the lowest amount of rainfall is present the dominance indexes were
the lowest and the species was classified as dominant or subdominant.


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Dispersal of click beetle and cranefly species across an agricultural landscape
Rod Blackshaw, Helen Hicks

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Natural infection of wireworm, Agriotes sp. (Coleoptera; Elateridae),
with Rickettsiella bacteria

Christina Schuster, Claudia Ritter, Regina G. Kleespies, Simon Feiertag, Andreas Leclerque

Abstract: Wireworms, the polyphagous larvae of click beetles belonging to the genus Agriotes
(Coleoptera: Elateridae), are severe and widespread agricultural pests affecting numerous crops
worldwide. A previously unknown intracellular bacterium has been identified in a diseased
Agriotes larva. Microscopic studies revealed the subcellular structures characteristic of
rickettsiosis. Molecular phylogenetic analysis based on 16S ribosomal RNA encoding rrs gene
demonstrated that the wireworm pathogen belongs to the taxonomic genus Rickettsiella
(Gammaproteobacteria; Legionellales). The pathotype designation `Rickettsiella agriotidis´ has
been proposed to refer to this organism. Moreover, genetic analysis makes it likely that this new
pathotype should be considered a synonym of the nomenclatural type species, Rickettsiella


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Potential control of Swiss wireworms with entomopathogenic fungi
Ursula Kölliker, Lorenzo Biasio, Werner Jossi

Abstract: The main wireworm species reducing the quality of potato tubers in northern
Switzerland are Agriotes obscurus, A. lineatus and A. sputator. In this study, the effect of the
Swiss Metarhizium anisopliae strain ART-2825 and of Naturalis® (Beauveria bassiana) on these
wireworm species was evaluated in the laboratory, in the greenhouse and in the field. In a
laboratory bioassay, larvae of the three Agriotes species treated with Naturalis® product and
Naturalis® spores did not exceed the infection rate of the control treatment. Similarly, no
significant effect of Naturalis® was observed in a potato field, with 79% percent damaged tubers
in the control and 82% in the Naturalis® treatment. In contrast, M. anisopliae strain ART-2825
demonstrated a high virulence towards A. obscurus in two laboratory bioassays. Nine weeks postinoculation,
80% and 97% infected A. obscurus larvae were observed in bioassay 1 and bioassay
2, respectively. However, the virulence of strain ART-2825 against A. lineatus and A. sputator
was considerably lower with maximum infection rates of 50%. Application of strain ART-2825
onto sterile and non-sterile soil in pots in the greenhouse resulted in a significant reduction of
surviving A. obscurus larvae. The corrected efficacy according to Abbott of strain ART-2825 was
61% in sterile and 50% in non-sterile soil. The results suggest that Naturalis® is not suitable to
control wireworms in potato fields in northern Switzerland. However, M. anisopliae strain ART-
2825 may be investigated further as a means to control wireworms in Swiss IPM programmes.


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Alternative methods to control wireworms (Agriotes spp., Coleoptera: Elateridae)
in vegetable production – potential of calcium cyanamide
and Metarhizium anisopliae

Claudia Ritter, Kai-Uwe Katroschan, Ellen Richter

Abstract: Wireworms of Agriotes spp. are severe and widespread agricultural pests affecting
especially maize, potatoes and numerous vegetable crops. Within the framework of a national
cooperation project on pest management strategies against soil insects, special emphasis was
placed on evaluating alternative wireworm control methods for vegetable production. Therefore,
the potential of calcium cyanamide and Metarhizium anisopliae was investigated. Calcium
cyanamide showed a repellent but no lethal effect on late instars of Agriotes ustulatus. A Swiss
Metarhizium anisopliae strain revealed promising crop protection effects requiring further
investigations concerning treatment reliability and commercialisation.


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Evaluation of four different bait traps to sample wireworms (Coleoptera: Elateridae)
infesting wheat and barley crops in Montana

Anuar Morales-Rodriguez, Aracely Ospina-Lopez and Kevin W. Wanner

Abstract: Wireworms are the most important soil insect pest of wheat and barley in Montana.
Baited pitfall traps have been used in several studies in North America for sampling wireworms
in soil. Wheat, wheat/corn mixtures, oatmeal, and grain sorghum seeds as well as pieces of carrot
and potato have been used as bait in underground pitfall traps. In general, wireworms were
equally attracted by all baits. Baited pitfall traps used to assess absence/presence of wireworms in
crops and grassland is a more labour efficient method compared to sampling soil cores. We
compared underground classical pitfall, pot, stocking and canister traps in spring wheat and
barley fields during the spring and summer of 2010. Captures made with the four pitfall traps
baited with a wheat/barley mixture, along with seasonal distribution of the different species that
were collected, will be summarized.


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Sampling for wireworms (Coleoptera: Elateridae) at sugarcane planting
Sampling for wireworms (Coleoptera: Elateridae) at sugarcane planting

Abstract: Wireworms in Florida are primarily a pest in newly planted sugarcane where the
larvae attack the underground portions of the plant by feeding on the buds and root primordia
during germination and on shoots and roots after germination. Florida sugarcane growers usually
apply a soil insecticide at planting to protect germinating seedpieces from wireworm damage.
However, studies have suggested that this application in many cases may not be necessary. The
objective of our research was to determine if a quick and easy sampling method could be used by
Florida sugarcane growers to determine the need for soil insecticide at planting. Testing was
conducted on fields located on King Ranch near South Bay, Florida. Wireworm samples were
taken in a transect across the length of each field. Sugarcane yield parameters for each whole
field were obtained through standard commercial harvesting procedures. Using our sampling
method to determine the necessity of soil insecticide application, yield data showed that there
was no significant difference in gross tons of sugarcane per ha, % sucrose, or tons of sugar per ha
in fields with versus without insecticide application.


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Dispersal of Agriotes beetles and their larvae revealed by stable isotope analysis
Nikolaus Schallhart, Manuel J. Tusch, Bettina Thalinger, Karin Staudacher, Corinna Wallinger, Anita Juen, Michael Traugott

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Agriotes species: Comparison of species composition in pheromone trap catches
with larval bait trap catches at the same site

Michèle Mangen, Marion Landl, Johann Glauninger

Abstract: In 2007 and 2008 several reportedly Agriotes infested sites in Lower and Upper
Austria were selected. At each site, bait traps were placed in the soil to check for the presence of
the agricultural harmful larvae. The larvae were determined with molecular methods; only
Agriotes ustulatus was determined with morphological methods. At the same sites pheromone
traps were placed and after 2 weeks their content was determined. The focus was on the five, in
literature described as most harmful, species in Austria (Agriotes ustulatus, Agriotes sputator,
Agriotes brevis, Agriotes obscurus
and Agriotes lineatus). Interestingly, Agriotes lineatus larvae
were very rare to totally absent in all the sites, which is contradictory to the adult catches. Due to
these findings, the assumption that pheromone trap catches reflect the larval population cannot be
made. Further studies analysing the origin of the beetles caught in the pheromone traps may help
to understand this discrepancy.


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Morphological traits of Agriotes obscurus and Agriotes ustulatus
(Coleoptera: Elateridae) larvae

Michèle Mangen, Rudoph Wegensteiner, Johann Glauninger

Abstract: From 2007 until 2009 Agriotes larvae were collected at several sites in Upper and
Lower Austria. These larvae were determined with molecular methods. As it is still a very
complicated task to determine the Agriotes larvae based on morphological traits, we decided to
have a closer look at the already reliably determined larvae under the scanning electron
microscope (SEM). Up until now, no pictures of the morphological traits have been published
and especially no pictures of individuals determined with molecular methods.


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Development of amplified fragment length polymorphism (AFLP) markers
for assessing Agriotes click beetle dispersal in agricultural land

Carly Benefer, Rod Blackshaw, Mairi Knight, Jon Ellis

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Ecology and control of wireworms in Florida sugarcane
Ron Cherry, Alvin Wilson

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Biodiversity of click beetles (Elateridae) in the agriculture landscape
of Saxony-Anhalt – Results of pheromone trap-monitoring

Tackenberg, Maria, Wolff, Christian, Volkmar, Christa, Lübke-Al Hussein, Marita

Abstract: Wireworms become more important as substantial pests in the agriculture landscape of
Saxony-Anhalt, because of the reinforced intercropping, the land set-aside, the greening, the
weed infestation, the ploughing up of grassland and the reduced cultivation (Vidal & Petersen
2010). In 2001 a similar Monitoring had taken place in France to proof if the occurrence of
Agriotes sordidus was spread. This species has a shortened cycle as the other ones, why the
injuries increased (Furlan 2004). That’s why a throughout Germany monitoring took place. Their
adult occurrence was evaluated by an Elateridae-Monitoring in Saxony-Anhalt in 2009 and 2010.
Concerning this we had pheromone traps of Furlan from Syngenta for the species Agriotes
lineatus, Agriotes obscurus, Agriotes sputator, Agriotes sordidus and Agriotes ustulatus. In 2009
the traps were positioned at 4 habitats of effort and in 2010 at 5 habitats of effort in the different
geographic natural environment in Saxony-Anhalt. The habitats were Poppau (Altmark),
Giesenslage (Altmarkkreis Stendal), Quedlinburg (Harz), Bornum (Anhalt-Zerbst) and
Dederstedt (Mansfeld-Südharz). The occurrence of Agriotes sordidus can’t be proved in both
years. The other species of Agriotes were found in different intensities at all habitats.


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Monitoring of click beetles with the use of pheromone traps in hop yards
of the Hallertau

Florian Weihrauch, Johannes Schwarz

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How plant identity and diversity affect food choices of Agriotes larvae
Nikolaus Schallhart, Manuel J. Tusch, Corinna Wallinger, Karin Staudacher, Michael Traugott

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Detection rates of ingested plant-DNA in Agriotes wireworms
Corinna Wallinger, Karin Staudacher, Nikolaus Schallhart, Anita Juen, Michael Traugott

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