IOBC-WPRS Shop: ePublications

View basket

IOBC-WPRS Bulletin Vol. 90, 2013

 

IOBC-WPRS Bulletin Vol. 90, 2013

Working Group "Insect Pathogens and Entomoparasitic Nematodes".
Preceedings of the meeting "Biological Control - its unique role in organic and integrated production" at Zagreb (Croatia), 16 - 20 June, 2013.
Edited by Johannes A. Jehle, Renata Bazok, Neil Crickmore, Miguel López-Ferber, Itamar Glazer, Enrique Quesada-Moraga, Michael Traugott.
ISBN 978-92-9067-268-5 [XXIII + 363 pp.]

 

25.00 €

 

 

 

 

Add this product to your basket

Articles

Pages

Price

Cart

Authorisation of biological control agents - theory and practice
Ralf-Udo Ehlers

Abstract: The legislation of the European Union regarding the placement of plant protection
products on the market (Regulation (EC) No 1107/2009) and also the Directive 2009/128/EC on
the sustainable use of pesticides pave the ground for increasing use of biological control agents in
Europe. Both clearly give priority to the use of alternative, non-chemical control measures.
However, the general practice in member states is different. Whereas member states largely seem
to ignore the potential of biological control, chemical control companies have increasing interest
in biological control and have acquired several biocontrol companies in order to get access to
biocontrol biodiversity and know-how.

3-7

5.00 €

 

Dynamics of baculovirus as insect biocontrol agent
Just M. Vlak, Monique M. van Oers

Extended abstract

9-10

0.00 €

 

Insect pathogenic fungi: what was obtained and where to go?
Jørgen Eilenberg

Abstract: Since its start in 1991 the IOBC working group ‘Insect Pathogens and Insect Parasitic
Nematodes’ has held 13 meetings, plus some subgroup meetings. Papers on fungal
entomopathogens have been presented at all meetings, and likewise, fungi have been part of
presentations at all Annual Meetings in Society for Invertebrate Pathology (SIP). By looking into
especially the most recent presentations and literature I will discuss the status of insect
pathogenic fungi in biological control: what was obtained and where to go?

11-13

5.00 €

 

Biological control of wireworms with entomopathogenic fungi
S. Eckard, M. A. Ansari, T. M. Butt, J. Enkerli, G. Grabenweger

Abstract: Three species of wireworms, Agriotes lineatus, A. obscurus and A. sputator, are
economically important soil pests in arable and vegetable crops in Europe. Fungi of the genus
Metarhizium are natural pathogens of wireworms. We tested the virulence of three European
Metarhizium strains in laboratory experiments and found a maximum mortality of up to 80% four
weeks post inoculation. We further investigated stability of the virulence of the most effective
strain ART2825 after ten times of subcultivation on artificial media. There was no difference in
virulence compared to a treatment of larvae that were infected with freshly host-passed conidia.
We conclude that Metarhizium strain ART2825 is a potential candidate for the control of
wireworms and we will continue to validate its efficacy under field conditions.

17-20

5.00 €

 

Monitoring of the entomopathogenic fungus Beauveria brongniartii in cockchafer infested areas of the Euroregion Tyrol
Johanna Mayerhofer, Jürg Enkerli, Roland Zelger, Hermann Strasser

Abstract: The persistence of the entomopathogenic fungus Beauveria brongniartii in soils in the
Euroregion Tyrol has been evaluated over a period of two decades. The fungal product
Melocont® Pilzgerste was successfully applied on the fields in different concentrations and
intervals between 1989 and 2012. In July and August 2012 the soil samples were drawn and
analyzed on selective media to determine the occurrence of Beauveria spp. Preliminary results
from microsatellite analysis showed that the re-isolated Beauveria strains from the test sites,
which had been treated during the last four years, were identified as the production strain. New
insights into colonisation, mobility and persistence of B. brongniartii in soils will be discussed in
the presentation.

21-25

5.00 €

 

Susceptibility of Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae) to entomopathogenic fungi:
Laboratory assays and field trials

Hannes Rauch, Roland Zelger, Stefan Hutwimmer, Hermann Strasser

Abstract: Since the introduction of Diabrotica virgifera virgifera to Europe between the late
1980s and the early 2000s, the western corn rootworm subsequently has extended its presence
across many parts of Europe and can currently be found in 20 European countries. Several
different strategies aiming at the control of D. v. virgifera have at least partially limitations,
making the biological control probably the most encouraging management method. Bioassays
revealed two Metarhizium anisopliae strains and one Beauveria bassiana strain with the highest
pathogenicity against 3rd instar larvae of D. v. virgifera. Although results obtained from fungal
density measurements after fungal application in Hungarian croplands revealed quite displeased
persistences, the potential of certain fungal strains for the control of the western corn rootworm
can be considered indisputable. However, further lab assays as well as field trials are needed to
confirm this high potential.

27-31

5.00 €

 

Efficacy of biological control agents for the control of western corn rootworm
Emese Balog, Bui Xuan Hung, György Turóczi, József Kiss

Abstract: The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, has been
introduced to Europe more than 20 years ago, and it is a well-established maize pest in Hungary
since 1995. The larvae of WCR cause damage on the maize roots. The efficacy of various
biological control agents (BCAs), such as fermented cultures of various entomopathogenic toxin
producing preparations of Bacillus thuringiensis, and some strains of the entomopathogenic
conidial fungus Metarhizium anisopliae, was screened against the larvae of WCR but the
practical application of them still needs additional research and development inputs. In in vitro
tests, WCR larvae were treated with microbial products (fermented cell cultures or spore
suspensions in various concentrations) at the second larval stage. Larvae were fed with freshly
germinated maize roots and larval mortality was recorded until pupation. In greenhouse
experiments maize plants were grown in pots placed in isolators. WCR eggs (20 for each plant)
were put directly under the seeds. In greenhouse experiments the microbial preparations were
applied at the time of sowing, in the same way as they were applied in the in vitro trials. One
month after the planting, the root mass was measured, and the damage caused by larvae was
determined based on the modified IOWA 1-6 scale. Most of the bacterial preparations and fungal
strains proved to be effective both in killing WCR larvae and preventing root damage on maize
plants. Some microbial treatments almost reached the efficacy of the control treatments
(Tefluthrin (FORCE 1.5 G) and Bacillus thuringiensis var. tenebrionis (NOVODOR FC)) and can
be considered as promising control agents of WCR.

33-36

5.00 €

 

Exploring synergistic effects of semiochemicals, entomopathogenic fungi and nematodes against root-herbivores
Michael A. Brandl, Mario Schumann, Stefan Vidal

Abstract only

37

0.00 €

 

Entomopathogenic fungi ecology and diversity from different Mediterranean ecosystems
María Fernández-Bravo, Enrique Quesada-Moraga, Inmaculada Garrido-Jurado

Abstract only

41

0.00 €

 

Efficacy of two strains of Beauveria bassiana entomopathogenic fungus on the red palm weevil in France and in Spain
Samantha Besse, Ludovic Crabos, Karine Panchaud

Abstract only

42

0.00 €

 

Beauveria bassiana strain ATCC 74040 interferes with oviposition behavior of Mediterranean fruit fly
Luca Ruiu, Giovanni Falchi, Edith Ladurner

Abstract: The entomopathogenic fungus Beauveria bassiana is known to interact with insects in
several ways. The present work reports the results of observations on the potential of Beauveria
bassiana strain ATCC 74040 against the Mediterranean fruit fly, Ceratitis capitata, with special
regard to disturbance effects on oviposition behaviour. A commercial formulation (Naturalis) and
different fungal preparations (pure conidia, hyphae, culture supernatants) were applied to orange
fruits offered to ovipositing medflies. A significantly lower number of fly visits and oviposition
punctures were recorded on fruits treated with Naturalis and with pure conidia than on control
fruits. The observed effects are examined on the basis of additional proteomic and genomic
observations, and the potential molecular implications of the rodlet layer of aerial conidia are
discussed.

43-46

5.00 €

 

Pathogenicity of an indigenous strain of the entomopathogenic fungus Beauveria bassiana on larvae and adults of the sisal weevil, Scyphophorus acupunctatus Gyllenhal (Coleoptera: Curculionidae)
V. T. Gkounti, D. Markoyiannaki, D. C. Kontodimas

Abstract: The sisal weevil is a severe pest of both ornamental and cultivated agave species. As
the use of synthetic insecticides causes undesirable effects, the evaluation of potential biological
control agents is necessary. Field collected adults and larvae of Scyphophorus acupunctatus were
used to evaluate the pathogenicity of an indigenous strain of the entomopathogenic fungus
Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales). Different concentrations
of spore suspensions were tested. As in some cases 100% mortality was achieved it is indicating
that this strain could serve as a potential biological control agent of the sisal weevil.

47-49

5.00 €

 

Microbial control of European red spider mite (Panonychus ulmi) with Beauveria bassiana strain ATCC 74040
Edith Ladurner, Massimo Benuzzi, Andrea Braggio, Sergio Franceschini, Veselin Zivkovic

Abstract: The European red spider mite, Panonychus ulmi, can cause severe damage on many
fruit crops, especially on apple. Outbreaks of mite populations usually occur in summer on warm
and humid days. Natural occurring predator populations may not always be able to keep the pest
under control, especially because of the likely occurrence of a lag in time in build-up of prey and
predator populations and due to the use of non-selective chemical pesticides. The efficacy of the
microbial control agent Beauveria bassiana strain ATCC 74040 (Naturalis®) against P. ulmi was
tested in open trials on apple. In one of the trials, also observations on the potential side effects of
the formulated product on natural occurring predator populations (Phytoseiulus spp. and
Stethorus punctillum) were made. The microbial control agent showed high efficacy against
P. ulmi in all field trials, and did not adversely affect predator populations. Beauveria bassiana
strain ATCC 74040 can be considered a valuable tool to be integrated into P. ulmi control
strategies.

51-55

5.00 €

 

Mycopathogens of the corn leaf aphid, Rhopalosiphum maidis (Fitch.) infesting wheat plants at Assiut, Egypt
Ahmed Y. Abdel-Mallek, Mohamed A. A. Abdel-Rahman, Gamal H. A. Hamam

Abstract only

56

0.00 €

 

Exploiting vine weevil behaviour to disseminate an entomopathogenic fungus
Tom Pope, Charlotte Arbona, Harriet Roberts, Jude Bennison, John Buxton, Gill Prince and Dave Chandler

Abstract: Control of adult vine weevil (Otiorhynchus sulcatus) is currently reliant on the use of
insecticides. However, using insecticide applications targeted against this pest is difficult, as they
need to be applied at dusk, and are often incompatible with integrated pest management
programmes. This study investigated the potential of a novel control strategy that uses artificial
refuges containing spores of an entomopathogenic fungus and exploits vine weevil behaviour to
disseminate the pathogen throughout weevil populations.
Preliminary experiments identified a simple plastic crawling insect trap as a suitable
artificial vine weevil refuge. Subsequent semi-field experiments using fluorescent powders in
place of an entomopathogenic fungus spore formulation showed that vine weevil aggregation
behaviour and movement between refuges effectively disseminated the powders throughout
weevil populations.

59-62

5.00 €

 

Field persistence of Metarhizium spp. strains applied as biocontrol agents against ticks (Ixodes ricinus)
Maria Mitteregger, Sarah Sonderegger, Hermann Strasser

Abstract: In two semi-field trials the persistence of three Metarhizium strains (BIPESCO 5,
ARSEF 3297, ARSEF 4556) after foliar spray application was monitored and bioassays with
Tenebrio molitor larvae were made to preclude a negative effect on germination and vitality of
conidia by using the adhesive agent Neo-wett™ and the antifoaming agent Antischiuma
Schaumstop™. Three different outdoor conditions (i.e. unprotected, rainfall protected, rainfall
protected and fully covered) were tested and all strains showed an adequate persistence after 25
days (20-50%). BIPESCO 5 conidia were re-isolated from foliage even after heavy rain showers
in open sites after 44 days. Conidia showed high virulence in bioassay with and without Neowett
™ and Antischiuma Schaumstop™: 90% of all larvae were killed after 5 to 20 days. No
negative effects of the adhesive- and antifoaming agent on the germination ability and vitality of
Metarhizium conidia were observed.

63-67

5.00 €

 

Vertical transmission of an endophytic strain of Beauveria bassiana (Ascomycota; Hypocreales) colonizing opium poppy Papaver somniferum
Enrique Quesada-Moraga, Blanca B. Landa del Castillo, Cristina López-Díaz

Abstract only

68

0.00 €

 

Development of a novel fermentation and formulation process for an endophytic Beauveria bassiana strain
Rieke Lohse, Desiree Jakobs-Schönwandt, Anant Patel

Abstract: There is an increasing demand for alternative or complementary crop protection
strategies. A novel approach could be the use of the entomopathogenic and endophytic fungus
Beauveria bassiana isolate ATP-04. To use the endophyte as a commercial biocontrol agent, the
fungus has to be mass-produced. B. bassiana was raised in shake flask cultures to produce
submerged conidiospores (SCS) which are reported to show a higher shelf life than mycelium
and blastospores (BS). It was found that in mineral media with 5% sugar beet molasses
B. bassiana produced 0.1 x 1010 SCS g-1 sucrose in 192 h. By adding 50 g l-1 NaCl 48 h after
inoculation the SCS yield increased to 1.4 x 1010 SCS g-1 sucrose. The scale-up to a 2 l stirred
tank reactor was carried out at 25°C, 200-600 rpm and 1 vvm at pH 5.5. A total spore yield of 5.2
x 1010 spores g-1 sucrose corresponding to a SCS yield of 0.2 x 1010 SCS g-1 sucrose was obtained
after 216 h. Also the yield of SCS increased to 1.1 x 1010 SCS g-1 sucrose by the addition of
NaCl. After fermentation the B. bassiana was formulated in a novel spray formulation that
delivers the fungus on oilseed rape leaves, increases persistence, germination and growth on
leaves as well as penetration, colonization and efficacy in bioassays with Plutella xylostella.

69-73

5.00 €

 

Development of analytical tools to monitor the fate of Metarhizium anisopliae metabolites in the environment
Judith Taibon, Sonja Sturm, Christoph Seger, Hermann Strasser, Hermann Stuppner

Abstract: Destruxins (dtxs) are structurally closely related cyclic hexadepsipeptides secreted as
relevant metabolites by the entomopathogenic fungus Metarhizium anisopliae. To monitor dtxs in
fungal culture broth, plant derived matrices and cell cultures, a fast and selective off-line SPE
UHPLC-DAD/MS method was established. Sample preparation was carried out by a solid phase
extraction (SPE) on a reversed phase material. Optimal purification was achieved by a washing
step with 40% (v/v) methanol, removing most of the polar components. The highest amounts of
dtxs were obtained by using 85% (v/v) methanol for elution. An UHPLC-DAD system
hyphenated to a Q-TOF mass spectrometer was utilized to separate and detect the dtx congeners.
A sub-2μm particle size column was used as stationary phase, with a water/acetonitrile solvent
gradient at a flow rate of 0.3 ml min-1 serving as mobile phase. A total analysis time of 12 min
was achieved with the UHPLC-DAD assay with the dtx congeners eluting from 1 min to 8 min
with a higher resolution of the peaks compared to previous HPLC-DAD assays. Besides the
available reference compounds dtxA, dtxB, dtxE, dtxE-diol 18 dtx derivatives were tentatively
identified by analyzing TOF-MS data.

75-78

5.00 €

 

Cross-species transferability of 41 microsatellite markers for Metarhizium spp.
Andy Lutz, Franco Widmer, Adrian Leuchtmann, Jürg Enkerli

Abstract: The genus Metarhizium includes insect pathogenic fungal species, which are used as
biological control agents (BCAs). Genetic tools for identification and monitoring of such BCAs
are important. A genotyping tool based on 41 simple sequence reapeat (SSR) markers has been
developed for M. anisopliae s.l. However, detailed phylogenetic analyses based on a multilocus
approach revealed that M. anisopliae s.l. is a cryptic species complex of nine different
Metarhizium species. According to this new taxonomy, the 41 SSR markers were isolated from
M. brunneum, M. robertsii or M. anisopliae s.s. The goal of this study was to assess the
transferability of the 41 SSR markers to individual species of the former M. anisopliae species
complex. Successful PCR-amplification of SSR markers was observed in all species but the
number of loci yielding PCR products varied among species. Amplification of individual SSR
loci did not always yield products for all strains of a particular species and not all were
polymorphic. The study revealed that SSR markers can be transferred to different species of the
former M. anisopliae species complex. However, the number of available SSR markers strongly
depends on the species to be analyzed. The markers will provide a valuable tool for identification
and monitoring of Metarhizium BCAs and they will allow investigation of genetic diversity and
population structure of seven species of the former M. anisopliae species complex.

79-83

5.00 €

 

A review of the use of entomopathogenic fungi for the control of Bemisia tabaci (Hemiptera: Aleyrodidae) in the UK
Andrew G. S. Cuthbertson

No abstract

87-90

5.00 €

 

Effect of entomopathogic fungi against Trialeurodes vaporariorum and its parasitoid Encarsia formosa: preliminary laboratory assays
Monica Oreste, Michele Poliseno, Eustachio Tarasco

Abstract only

91

0.00 €

 

Laboratory Beauveria bassiana (Bals.) Vuill. bioassays on spruce bark beetle (Ips typographus L.)
Ana-Maria Andrei, Daniela Lupăştean, Constantin Ciornei, Ana-Cristina Fătu, Mihaela Monica Dinu

Abstract: The massive damages caused by Ips typographus in spruce forests in Romania, the
severe restrictions on the use of chemical insecticides, the identification of natural Beauveria
bassiana outbreaks in the Romanian forests infested with bark beetle and the isolation of a new
B. bassiana strain, led to the development of some researches on the possibility to use this
entomopathogenic fungi for reducing the damage caused by bark beetles. In laboratory
conditions, the susceptibility of I. typographus to infection by a naturally occurring B. bassiana
strain was tested. Utilization of a B. bassiana conidial suspension (3.31 x 1011 conidia ml-1)
induced beetle mortality, length of mother galleries reduction and larval galleries number
reduction.

93-96

5.00 €

 

Effect of local strains of Beauveria bassiana (Bb024) and Metarhizium anisopliae (M7/2) against the fallweb worm Hyphantria cunea (Lepidoptera: Arctiidae) in Georgia
Medea Burjanadze, Elena Nakaidze, Mariam Arjevanidze, Tea Abramishvili

Abstract: The indigenous isolates of the entomopathogenic fungi Beauveria bassiana (Bb-024)
and Metarhizium anisopliae (M7/2) against fifth to seventh instars larvae of Hyphantria cunea in
two concentrations of 1 x 107 and 1 x 108 conidia ml-1 were tested in the laboratory. Maximum
mortality of larvae was observed 4-9 d after treatment. Both isolates were pathogenic to H. cunea
larvae. The mean mortality caused by B. bassiana ranged from 59.8% to 84.3% and that of
M. anisopliae ranged from 52% to 68%. The living larvae hidden under leaves and cordon made
cocoons and transformed into pupae. The adult moths appeared from the overwintering pupae.
They began to emerge massively and mated. The emergence of adults of B. bassiana was 69.6%,
that of M. anisopliae was 60%, and that of the control was 55.7%. The larvae hatched 7 to 10 d
later (the hatching rates were: B. bassiana – 76.3%, M. anisopliae – 70%, control – 89.5%). In
case of B. bassiana treatment, dimorphic males emerged from pupae often showing undeveloped
wings.

97-101

5.00 €

 

Highly effective Beauveria pseudobassiana strain (Dm-5) against the great spruce bark beetle, Dendroctonus micans (Kugelann) (Coleoptera: Scolytidae)
Ismail Demir, Seda Kocacevik, Ali Sevim, Mahmut Eroglu, Zihni Demirbag

Abstract only

102

0.00 €

 

Laboratory testing of insect associated fungi for the control of wireworms (Agriotes sp. L.)
Jaka Razinger, Matthias Lutz, Hans-Josef Schroers, Gregor Urek, Jürg Grunder

Abstract: The aim of the study was to assess entomopathogenic potential of 7 isolates from 6
entomopathogenic fungal species (EPF) isolated from various substrats in Slovenia against larvae
of Agriotes sp. The fungal isolates tested were Beauveria bassiana, B. brongniartii, Metarhizium
anisopliae (2 isolates), M. robertsii, Purpureocillium lilacinum and Clonostachys solani. Conidia
of these species were incorporated into the test substrate as a water suspension to reach a final
concentration of 3.85 x 106 conidia g-1 air-dried soil. The larval mortality was observed on a
weekly basis for a total of 90 days. The mortalities observed exhibited a linear trend with slopes
ranging from 0.20 to 1.23 for the fungal treatments and 0.08 to 0.18 for the control treatments.
Abbott’s corrected mortality at day 90 ranged from 20.7 to 76.9%. The most promising candidate
biological control agent was Metarhizium anisopliae isolate 1154.

103-107

5.00 €

 

Laboratory and semi-field trials on the effects of Beauveria bassiana (JW-1, ATCC 74040) against soil-dwelling stages
of Frankliniella occidentalis (Thysanoptera: Thripidae)

Andrea Boaria, Alberto Pozzebon, Mauro Pesce, Mauro Lorenzon, Carlo Duso

Abstract: Beauveria bassiana (Balsamo) Vuill. is an entomopathogenic fungus used in
controlling various pests. Previous research showed that B. bassiana, applied to the plant canopy,
could exert a significant control of thrips populations, in particular of Frankliniella occidentalis
Pergande. However, some stages (e.g., prepupae and pupae) of this species develop in the soil
being less affected by control treatments applied to the canopy. The identification of biological
control agents active against soil-dwelling stages of F. occidentalis is an important issue for the
implementation of IPM. Here we present laboratory and greenhouse experiments carried out to
evaluate the potential of B. bassiana (JW-1 ATCC 74040) in controlling soil-dwelling stages of
F. occidentalis. In laboratory bioassays B. bassiana reduced significantly the emergence of
F. occidentalis adults. In the greenhouse experiment, a significant control of thrips population
was obtained on cyclamen potted plants.

109-112

5.00 €

 

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 ef1a 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.

113-117

5.00 €

 

Evaluation of indigenous Beauveria isolates as potential agents for emerald ash borer management and the development
of a diagnostic marker to monitor a post-release isolate

George Kyei-Poku, Shajahan Johny

Abstract: To search for effective and safe indigenous biocontrol agents to manage emerald ash
borer (EAB), we conducted a survey in 2008-2009 of entomopathogenic fungi (EPF) infecting
EAB in outbreak sites in southwestern Ontario, Canada. Many Beauveria spp. isolates were
recovered from dead and mycosed EAB cadavers residing in the phloem tissues of dead ash
barks, larval frass extracted from feeding galleries under the bark of dead trees. Molecular
characterization using sequences of the ITS, 5' end of elongation factor 1 alpha (EF1-α) and
intergenic Bloc region fragments revealed that Beauveria bassiana and B. pseudobassiana were
commonly associated with EAB in the sampled sites. Initial virulence screening against EAB
adults of 23 isolates representing the different clades yielded 8 isolates that produced more than
90% mortality in a single concentration assay. These isolates differed in virulence based on LC50
values estimated from multiple concentration bioassay and based on mean survival times at a
conidia concentration of 2 × 106 conidia ml-1. B. bassiana isolate L49-1AA was significantly
more virulent and produced more conidia on EAB cadavers compared to the other indigenous
isolates and the commercial strain B. bassiana GHA suggesting that L49-1AA may have potential
as a control agent against EAB. Studies have been developed to use auto-contamination trapping
system to disseminate L49-1AA to manage EAB field populations. We targeted the EF1-α gene
sequence from L49-1AA to develop an allele/strain specific primer set that will be used to
monitor the introduced L49-1AA in terms of its establishment, persistence and virulence in the
environment.

119-124

5.00 €

 

Isolation and identification of endophytic entomopathogenic fungi from dent corn
Daigo Aiuchi, Tatsumi Takanami, Sayaka Toba, Minehiro Ishii, Shin-ichiro Asano, Masanori Koike

Abstract: In this study, we sought endophytic entomopathogenic fungi from dent corn, which
have potential to be developed as endophytic biopesticide with multiple roles. Dent corn samples
applied to this study were collected at three locations of East Hokkaido island. Each plant sample
was divided into root, stem, leaf and kernel, and then these were surface-sterilized by 70%
ethanol and 0.5% sodium hypochlorite. Pieces of each tissue were placed on entomopathogenic
fungi selective medium. All fungal isolates growing on this plate were transferred onto potato
dextrose agar. Morphological identification to genus level was conducted by slide culture method
by observing under light microscope (x100). In total, 2252 fungal isolates (greater part of isolates
were Penicillium spp. and Cladosporium spp.) were detected on selective medium, and among
them, 168 isolates were entomogenous fungi. Five genera of entomopathogenic fungi including
Beauveria, Lecanicillium, Isaria, Metarhizium and Simplicillium were detected in this study. In
this study, only five plant samples were applied, but entomopathogenic fungi were detected from
all locations and at all parts of plant tissue. Moreover, it is indicated that endophytic
entomopathogenic fungi multiply colonize in the plant body. Although, Beauveria, Lecanicillium,
Isaria and Metarhizium showed tendency to localize to some plant part, Simplicillium tended to
be ubiquitous presence in plant body. Our result can indicate that entomopathogenic fungi
universally colonize into dent corn.

125-128

5.00 €

 

Endophytic establishment of the entomopathogen Beauveria bassiana in Vitis vinifera plants
Yvonne Rondot, Annette Reineke

Abstract only

129

0.00 €

 

Effect of temperature, water activity and UV-B radiation on conidia germination and colony growth of Beauveria bassiana isolates from soil and phylloplane
María Fernández-Bravo, Inmaculada Garrido-Jurado, Monica Oreste, Enrique Quesada-Moraga

Abstract only

130

0.00 €

 

Deletion genotypes influence occlusion body potency and production in insects infected by a Spodoptera frugiperda nucleopolyhedrovirus isolate from Colombia
Gloria Barrera, Trevor Williams, Laura Villamizar, Primitivo Caballero, Oihane Simón

Abstract: The Colombian field isolate (SfCOL-wt) of Spodoptera frugiperda multiple
nucleopolyhedrovirus (SfMNPV) is a mixture of different genotypes. To evaluate the insecticidal
properties of the different variants in SfCOL-wt a plaque assay was performed and ten distinct
genotypes were identified. Genotype SfCOL-A was the most prevalent (71%) and showed a PstI
restriction profile identical to that of SfCOL-wt. The remaining nine genotypes presented
genomic deletions of 3.8-21.8 Kb that affected the region between open reading frames (ORFs)
sf20 and sf33. The potency of SfCOL-A occlusion bodies (OBs) was approximately 4-fold higher
than SfCOL-wt OBs, whereas the speed of kill of SfCOL-A was similar to that of SfCOL-wt.
Deletion genotype OBs were similarly or less potent than SfCOL-wt, but six deletion genotypes
were faster killing than SfCOL-wt. The potency of mixtures of OBs and co-occluded mixed
genotype OBs were consistently reduced in two-genotype mixtures involving equal proportions
of SfCOL-A and one of three deletion genotypes (SfCOL-C, -D or -F). Speed of kill and OB
production were improved only when certain genotype mixtures were co-occluded, although OB
production was higher in the SfCOL-wt isolate than in any of the genotypes or genotype mixtures
that we tested. The SfCOL-wt population appears to be structured to maximize the production of
OBs in each infected host suggesting this to be the principal limitation to transmission.

133-136

5.00 €

 

On the role of baculovirus photolyases in DNA repair upon UV damage of occlusion bodies
Magdalena A. Biernat, Primitivo Caballero, Just M. Vlak, Monique M. van Oers

Abstract: The use of baculoviruses in insect biocontrol is hampered by their sensitivity to
ultraviolet (UV) light. This irradiation induces cyclobutane pyrimidine dimers (CPDs) in DNA.
CPD-photolyases repair CPDs using visible light. Plusiine baculoviruses encode photolyases,
which could potentially repair UV-damage prior to infection of larvae. Whether the photolyases
encoded by Chrysodeixis chalcites nucleoplyhedrovirus are involved in UV damage repair was
tested by infecting larvae with UV-irradiated viral occlusion bodies (OBs) that were subsequently
treated with visible light or kept in the dark. The observed mortality was the same for both
treatments. We postulate that photolyases are not active as DNA repair enzymes in OBs, but may
play a role in other aspects of baculovirus pathogenesis.

137-142

5.00 €

 

Effect of top spray drying and freeze drying on the photostability and insecticidal activity of a Spodoptera frugiperda nucleopolyhedrovirus (SfMNPV 003) formulation
Mauricio Cruz, Martha Liliana Chaparro, Laura Fernanda Villamizar, Martha Isabel Gomez

Abstract: The nucleopolyhedroviruses are considered as an effective biopesticide against the fall
armyworm Spodoptera frugiperda. Top spray and freeze drying methods were used to prepare a
wettable powder formulation based on nucleopolyhedrovirus of S. frugiperda (SfMNPV 003) and
its photostability and virulence were assessed. Top spray drying method was more efficient for
removing moisture content than freeze drying. No obvious differences in the insecticidal
activities were observed for both drying methods although a higher photostability (88.54%) was
observed in the formulation prepared with top spray drying method compared to freeze drying
(77.77%) and unformulated virus (15.62%) after 6 hours of UV radiation exposure. Top spray
drying method was selected as the most favorable process for being implemented in a
manufacture process.

143-147

5.00 €

 

Variations in the susceptibility to CpGV in populations of the codling moth, Cydia pomonella
Benoît Graillot, Christine Blachere, Samantha Besse, Myriam Siegwart, Miguel López-Ferber

Abstract: Failure in codling moth populations control with CpGV in apple orchards has been
attributed to the action of a single allele located in the Z chromosome. However, differences in
the mortality patterns between genetically homogeneous susceptible and resistant insects in the
laboratory strongly suggest that other mechanisms are responsible of variation in the
susceptibility of insect to CpGV isolates.

149-153

5.00 €

 

Characterisation of novel CrleGV isolates for false codling moth control - lessons learnt from codling moth resistance to CpGV
John Opoku-Debrah, Sean Moore, Martin Hill, Caroline Knox

Abstract: Recently some codling moth, Cydia pomonella, populations in Europe developed
resistance to CpGV. In order to prepare for the possibility of a similar occurrence with the false
codling moth, Thaumatotibia leucotreta, in South Africa, a search was conducted for novel
CrleGV isolates. Through overcrowding, outbreaks of novel isolates were recorded from
laboratory populations of five geographically distinct host populations. The genetic novelty of
these and two commercially available isolates was confirmed through restriction enzyme analysis
and sequence analysis of the granulin and egt genes. Phylogenetic analysis showed the existence
of two CrleGV-SA genome types. Significant differences in virulence were also shown between
certain isolates against certain host populations.

155-159

5.00 €

 

Elucidation of a novel mode of resistance of codling moth against Cydia pomonella granulovirus by homogenization experiments
Annette J. Sauer, Eva Fritsch, Karin Undorf-Spahn, Johannes A. Jehle

Abstract: Since 2005, codling moth (CM, Cydia pomonella) populations with a reduced
susceptibility to Cydia pomonella granulovirus (CpGV, Baculoviridae) products have been
reported from about 40 European orchards. The resistance could be traced back to a single,
dominant, sex-linked gene. Currently, resistance management strategies are based on the
application of improved CpGV products containing resistance-overcoming CpGV isolates.
Recently, two CM field populations (NRW-WE and SA-GO) with a reduced susceptibility to
even these improved CpGV products were found. First single pair crossing experiments between
individuals of these resistant field colonies and a susceptible laboratory CM strain (CpS)
indicated that the inheritance of resistance of these populations did not follow the previously
described pattern of Z-linked, dominant resistance. In single-dose bioassays the susceptibility of
neonates of the resistant CM colonies NRW-WE and SA-GO to different CpGV isolates (CpGVM,
-S, -V15 and -E2) was estimated. The aim of the current study was the genetic
homogenization of the genetically heterogeneous field populations NRW-WE and SA-GO by two
different methods: (i) repeated single pair crossings followed by family selection and (ii)
successive mass crossing experiments under virus pressure. The resulting homogenous strains of
NRW-WE and SA-GO with fixed resistance will be used for backcrossing experiments with CpS
to elucidate the mode of inheritance of their resistance.

161-165

5.00 €

 

Biological control of the box tree moth Cydalima perspectalis with Anagrapha falcifera nucleopolyhedrovirus (AnfaNPV)
Jana Rose, Johannes A. Jehle, Regina G. Kleespies

Abstract: The box tree moth Cydalima perspectalis originated from East Asia. Since several
years, it is a novel invasive insect pest in many European countries, causing widespread damage
on box tree plants. The potential of the baculovirus Anagrapha falcifera nucleopolyhedrovirus
(AnfaNPV) as a potential biological control agent for the control of C. perspectalis was
investigated in this study. Two AnfaNPV isolates, termed Dn10 and BI-235, were used. The
infectivity of AnfaNPV Dn10 and BI-235 to C. perspectalis was evaluated by leaf disc bioassays
and the median lethal concentration (LC50) was determined for both isolates. In addition, light
and electron microscopic analyses were performed to study the infection process. In conclusion,
larvae of C. perspectalis were shown to be susceptible to both AnfaNPV isolates.

169-172

5.00 €

 

Interactions between structural proteins of Chilo iridescent virus
Emine Özşahin, Remziye Nalcacioglu, Just M. Vlak, Monique M. van Oers, Zihni Demirbağ

Abstract only

173

0.00 €

 

Natural populations of Spodoptera exigua are infected by multiple viruses: implications for the production and use of virus insecticides
Cristina Virto, David Navarro, M. Mar Tellez, Salvador Herrero, Trevor Williams, Rosa Murillo, Primitivo Caballero

Abstract: Covert infections of Spodoptera exigua multiple nucleopoliedrovirus (SeMNPV) have
been detected in laboratory or field populations of the homologous host, S. exigua. Two RNA
viruses belonging to the Iflaviridae family (SeIV-1, SeIV-2) were identified in transcriptome
studies from different laboratory colonies of S. exigua. The three viruses are vertically transmited
and establish persistent infections. For this reason, coinfection of individual insects by these
viruses is considered likely. In this study, we determined the prevalence of covert infections
caused by iflaviruses and SeMNPV in order to identify virus associations in natural S. exigua
populations. SeMNPV was detected in the 54% of field-caught adults, whereas 13% and 8% of
insects were infected by SeIV-1 and SeIV-2, respectively. The prevalence of SeIV-1 and SeIV-2
in F1 adults obtained in laboratory showed higher levels than in the parental generation, whereas
the prevalence of SeMNPV decreased from parents to their offspring. These findings have
important implications for the in vivo production of virus based insecticides using mass-reared
insects and the efficacy of these products in controling pest populations that may be coinfected
by iflaviruses.

175-177

5.00 €

 

Estimating the importance of maternal and paternal contributions to the vertical transmission of Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV)
Cristina Virto, Carlos A. Zárate, Rosa Murillo, Primitivo Caballero, Trevor Williams

Abstract: Vertical transmission of Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV)
is believed to be a common feature in field populations of S. exigua. To assess whether gender
affects trans-generational virus transmission, four mating groups were performed using healthy
and sublethally infected insects: i) healthy males (H♂) × healthy females (H♀); ii) infected males
(I♂) × healthy females (H♀); iii) healthy males (H♂) × infected females (I♀) and iv) infected
males (I♂) × infected females (I♀). These adults and their offspring were analyzed by qPCR to
detect SeMNPV infection. Both males and females were able to transmit the infection to the next
generation, although females infected a higher percentage of the offspring and female-mediated
transmission was more consistent than that of males. Venereal transmission appeared to be half as
effective as maternally-mediated transmission, and the main route of transmission is likely
transovarial rather than transovum. The prevalence of the infection in the offspring did not vary
according to gender, therefore both males and females can be infected by their parents in similar
proportions. Incorporating vertically-transmitted genotypes in biological insecticides might have
the potential for reducing pest densities and extending periods between virus applications.

179-181

5.00 €

 

Baculoviruses for the biological control of cutworms (Agrotis spp.)
Jörg T. Wennmann, Gianpiero Gueli Alletti, Johannes A. Jehle

Abstract: Caterpillars of the common cutworm Agrotis segetum and black cutworm A. ipsilon
(Lepidoptera: Noctuidae) are wasteful feeders of various crops in agriculture. These cutworms
are mainly controlled by chemical pesticides but recent attempts are aimed to control these soil
pests by the application of baculoviruses. Four different baculoviruses, namely Agrotis segetum
nucleopolyhedrovirus A (AgseNPV-A), Agrotis segetum nucleopolyhedrovirus B (AgseNPV-B),
Agrotis ipsilon multiple nucleopolyhedrovirus (AgipMNPV) and Agrotis segetum granulovirus
(AgseGV), were isolated from larvae of A. segetum and A. ipsilon and are considered as potential
biocontrol agents. In natural infections, larvae of both hosts are susceptible to all four viruses and
individual caterpillars of the common cutworm were observed to become in infected
simultaneously by AgseNPV-B and AgseGV. Co-infections may be advantageous in terms of
virulence and resistance management, although the level of interaction is critical. To test for a
mutualistic, neutral or antagonistic interaction and to evaluate a combined application of Agrotis-specific
baculoviruses, we exemplary performed mixed infection experiments of A. segetum
larvae that were exposed to AgseNPV-B and AgseGVat different concentration. For quantitative
analysis of the outcome of mixed infections as well as for quality control in virus production a
reliable method for detection and discriminative quantification for Agrotis-specific baculoviruses
is required. We established a multiplex PCR analysis based on highly specific oligonucleotides
which also permit quantification by quantitative PCR. As a prerequisite of these studies the
genome of AgseNPV-B was completely sequenced by 454 sequencing technique. Comparative
genome sequence analyses gave a detailed insight into the molecular setup of the three Agrotis-specific
NPVs and confirmed that they can be regarded as three different but close related
species. Our results will help to develop and evaluate Agrotis-specific baculoviruses as biocontrol
agents and to understand the evolutionary co-existence of viruses that are highly adapted to the
same hosts.

183-186

5.00 €

 

Insecticidal activity of a spray dried formulation based on a Colombian Spodoptera frugiperda nucleopolyhedrovirus
Judith Elena Camacho, Martha Isabel Gómez, Mauricio Cruz, Laura Fernanda Villamizar

Abstract: A Colombian Spodoptera frugiperda nucleopolyhedrovirus (SfMNPV 003) with high
potential for the development of an efficient biopesticide was microencapsulated by top spray
drying with a pH dependent polymer (Eudragit®S100) and its insecticidal activity was evaluated
under laboratory and greenhouse conditions. Significant differences between LC50 values of the
microencapsulated virus, the dried virus and the virus without any treatment were not detected
under laboratory conditions, suggesting that microencapsulation by top spray drying did not
affect the virus insecticidal activity. Three different microencapsulated batches showed the same
efficacy under greenhouse conditions and significant differences between formulated and
unformulated virus were not detected (p > 0.05). In conclusion, SfMNPV003 insecticidal activity
was not affected by formulation process and developed biopesticide demonstrated its potential for
S. frugiperda control and could be included in programs of integrated pest management (IPM).

189-193

5.00 €

 

Cydia pomonella granulovirus knockout mutants: The potential role of pe38 in overcoming codling moth resistance
Manuela Gebhardt, Karolin E. Eberle, Johannes A. Jehle

Abstract only

194

0.00 €

 

Sequence analysis of CpGV-R5 isolate, able to efficiently control CpGV-M resistant insects: relation between biological activity and genome
Benoît Graillot, Samantha Besse, Christine Blachère-Lopez, Jérôme Olivares, Myriam Siegwart, Miguel López-Ferber

Abstract: The CpGV-R5 isolate is able to overcome resistant populations of codling moth to the
CpGV-M isolate. The complete sequences of CpGV-R5 and the CpGV-M used for industrial
production at Natural Plant Protection have been determined. Among the differences found, some
are specific to the R5 isolate, others are common to various isolates able to overcome the
resistance, like modification on the p38 gene product.

195-199

5.00 €

 

Functional characterization of serine/threonine protein kinase gene (AMV197) of Amsacta moorei entomopoxvirus
Hacer Muratoglu, Remziye Nalcacioglu, Srini Perera, Basil Arif, Zihni Demirbag

Abstract only

201

0.00 €

 

Transcriptional analysis of CpGV isolates in Cydia molesta
Dönüs Toy, Diana Schneider, Zihni Demirbag, Johannes A. Jehle

Abstract only

202

0.00 €

 

An examination of stress-related activation of SeMNPV in covertly infected Spodoptera exigua
Cristina Virto, David Navarro, María Mar Tellez, Rosa Murillo, Trevor Williams, Primitivo Caballero

Abstract: The aim of this study was to evaluate the effect of different stress factors on covertly
infected Spodoptera exigua larvae in terms of nucleopolyhedrovirus (NPV) activation. For this,
adult survivors that had ingested occlusion bodies of S. exigua multiple nucleopolyhedrovirus
(SeMNPV) were mated and the subsequent generation (F1) tested for virus activation in the
second instar in both laboratory and field conditions. In the laboratory, a number of treatments
were tested including chemical stressors, inoculation with heterologous NPV species and Bacillus
thuringiensis spores and crystals. Both, parental and F1 adults were confirmed to harbor the
infection by qPCR. Virus activation was observed in insects treated with 0.1% copper sulphate,
1% iron sulphate, and 1 ppm sodium selenite, resuling in 12%, 15%, and 41% mortality due to
SeMNPV, respectively, whereas no larvae with symptoms of viral infection were registered in
virus-free controls. No effect on NPV-induced mortality was detected after inoculation with
heterologous virus. Field trials were carried out by artificial infestation of pepper crops in
experimental greenhouses using sublethally infected S. exigua larvae to evaluate copper sulfate
and sodium selenite as activation factors. Very little NPV-induced mortality (< 5%) was observed
in those larvae treated in field conditions.

203-205

5.00 €

 

Functional analysis of Chilo iridescent virus zinc-binding matrix metalloproteinase gene
Aydın Yesilyurt, Hacer Muratoglu, Zihni Demirbag, Remziye Nalcacioglu

Abstract only

206

0.00 €

 

Latest field results on the biological control of Diabrotica virgifera virgifera with nematodes
Ralf-Udo Ehlers

Abstract only

209

0.00 €

 

Development of new formulations for soil pest control
Miriam Hanitzsch, Michael Przyklenk, Bianca Pelzer, Anant Patel

Abstract: In formulation science, there are few systematic investigations on encapsulation of
agro-biologicals with regard to materials, methods and technology for mass production. The aim
of this work was to develop novel mechanically stable capsule systems with increased persistence
in soil. To this end, we tested different methods with several biopolymers, combinations of
biopolymers and lignin as capsule additive. Capsule systems were prepared by ionic gelation,
thermal gelation, complex coacervation and additional bead coating. In selected capsules, lignin
was incorporated. Capsules based on single biopolymers were able to form stable spherical
capsules, e.g. alginate, pectin derivates and gelatin. Capsules based on combinations of polymers
also showed stable capsule formation, e.g. alginate/gelatin, alginate/lignin and SEC/PDADMAC.
Additionally, lignin was used successfully as additive in SEC hollow beads. First experiments
indicate significant differences in biological degradability and thus persistence in soil with
different capsule systems. These novel capsule systems with increased persistence are suitable for
delivery of BCAs into the soil.

211-215

5.00 €

 

Click beetles disperse widely across farmland: what else do we need to know?
Rod Blackshaw, Robert S. Vernon

Abstract: For many years wireworm research focussed on the distribution of larvae and their
control in field crops. The development of sex pheromone lures has facilitated the study of adult
male distributions over greater spatial scales but the assumption that these would provide a
simpler monitoring technique has not been fulfilled and it is now clear that we cannot be
confident about what the trap counts actually mean. This secondary focus on adult males neglects
the critical issue of female behaviours in agricultural landscapes. Understanding these is essential
to the development of new management strategies in an era of declining insecticide availability.
In this paper we briefly review what is known about the movement of click beetles across
farmland and identify research gaps that need to be filled if we are to develop area-wide
management strategies.

217-220

5.00 €

 

Distribution and abundance of Agriotes ustulatus L. adults on pheromone traps in four regions in Croatia
Antonela Kozina, Maja Čačija, Renata Bažok

Abstract: During several years of investigations, the distribution and the abundance of Agriotes
ustulatus in four different regions of Croatia were researched with the aim to correlate the
abundance with the prevailed climatic conditions in each region. Agriotes ustulatus was captured
by pheromone traps (Csalomon) on 17 fields distributed at seven localities in four different
regions in Croatia according to the climatic data. The highest dominance indices of A. ustulatus
were recorded in the warmest county, County of Vukovar-Srijem and species was classified as
eudominant. Agriotes ustulatus was subdominant at locality Čazma where the average
temperature was the lowest comparing to the other localities.

221-225

5.00 €

 

Efforts to develop female-targeted attractants for click beetles – a summary
Miklós Tóth, Lorenzo Furlan, József Vuts, Éva Bálintné Csonka, István Szarukán, Teodora B. Toshova, Mitko Subchev, Dimitar I. Velchev, Christine M. Woodcock, John C. Caulfield, Patrick Mayon, John A. Pickett, Michael A. Birkett

Abstract: An overview is given on recent research efforts to develop attractant combinations
capable of attracting female click beetles.

227-230

5.00 €

 

New perspectives for wireworm control based on an improved understanding of their feeding ecology
Michael Traugott, Karin Staudacher, Nikolaus Schallhart, Corinna Wallinger

Abstract only

231

0.00 €

 

Exploratory use of geometric morphometrics in the identification of wireworm species
Darija Lemić, Katarina Mikac, Hugo A. Benitez, Maja Čačija, Antonela Kozina, Renata Bažok

Abstract only

235

0.00 €

 

Development of novel biocontrol encapsulation techniques for garlic extracts: first results
Bianca Pelzer, Miriam Hanitzsch, Anant Patel

Abstract: The aim of the work presented here is to develop novel eco-efficient encapsulation
techniques for bioactive ingredients used in biological pest control for the EU project
INBIOSOIL. A process for production of small-scale alginate beads containing garlic extract by
air atomization with self-constructed technical encapsulation equipment was developed. The
encapsulation protects the active ingredients against oxygen and other outside influences, thus
enhancing shelf life and allows a slow release effect. The capsule size produced with this
technology can be varied – depending on the desired product – between 4 and 600 μm. Particles
are stable and spherical.

237-240

5.00 €

 

The project ATTRACT: Protection of crops from soil-borne insect pests with a novel attract and kill strategy
Marina Vemmer, Wilhelm Beitzen-Heineke, Hubertus Kleeberg, Edmund Hummel, Stefan Vidal, Anant Patel

Abstract: The project ATTRACT targets the development of a novel attract-and-kill strategy for
the protection of crops from soil-borne insect pests. The aim is the design of a plant protection
product with an innovative formulation based on CO2 emitting sources as an attractive compound
and environmentally friendly insecticidal compounds.

241-242

5.00 €

 

Combined use of entomopathogenic fungi and their extracts to improve the control of the cotton leafworm Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae)
Inmaculada Garrido-Jurado, Gloria Resquín-Romero, Enrique Quesada-Moraga

Abstract only

245

0.00 €

 

Insecticidal activity of a semi-purified extract from Metarhizium brunneum (Ascomycota: Clavicipitaceae) against the red palm weevil Rhynchophorus ferrugineus (Coleoptera: Curculionidae)
Inmaculada Garrido-Jurado, Óscar Dembilio, Josep Anton Jacas, Lola Ortega, Carlos Campos, Enrique Quesada-Moraga

Abstract only

246

0.00 €

 

Subterranean control of an arboreal pest: EPNs and EPFs for FCM
Sean Moore, Candice Coombes, Aruna Manrakhan, Wayne Kirkman, Martin Hill, Ralf-Udo Ehlers, John-Henry Daneel, Jeanne de Waal, Jo Dames, Antoinette Malan

Abstract: Control measures against the false codling moth (FCM), Thaumatotibia leucotreta,
have traditionally ignored the soil-borne pupal stage. Recent trials with entomopathogenic
nematodes (EPNs) and entomopathogenic fungi (EPFs) have targeted this life-stage. Application
of Heterorhabditis bacteriophora to a citrus orchard floor, reduced T. leucotreta infestation of
fruit by up to 81%. Conservation of H. zealandica through non-usage of a nematicide also
resulted in dramatically lower fruit infestation. Dose-response and exposure time-response
bioassays identified the three most promising fungal isolates against pupating T. leucotreta.
Orchard trials showed persistence of these fungi in orchard soil for at least six months.

247-250

5.00 €

 

Do plant-associated insect toxin producing pseudomonads have the potential for the biocontrol of insect pests?
M. Maurhofer, B. Ruffner, P. Flury, M. Péchy-Tarr, E. Fischer, P. Kupferschmied, C. Keel

Abstract only

251

0.00 €

 

Untangling insect pathogenicity in plant-associated pseudomonads by a combination of comparative genomics and bioassays
P. Flury, B. Ruffner, M. Péchy-Tarr, P. Kupferschmied, C. Keel, M. Maurhofer

Abstract only

252

0.00 €

 

Colorado potato beetle (Leptinotarsa decemlineata Say) – control strategies in organic farming using biological insecticides (azadirachtin, Bacillus thuringiensis var. tenebrionis, pyrethrum and spinosad)
Stefan Kühne, Uta Priegnitz, Benjamin Hummel, Frank Ellmer

Abstract: Field experiments using different control agents for Colarado potato beetle control
showed that all tested application strategies were effective. Time-shifted application of neem
(NeemAzal T/S) and Bacillus thuringiensis var. tenebrionis (B.t.t.) (Novodor FC) as well as
double treatment with B.t.t. achieved under optimal weather conditions gave an effectiveness
level of over 80% and increased yields. Surprisingly, a single application of spinosad (SpinTor)
also proved to be very effective (> 80%) in the three years studied in spite of the difficult study
conditions in 2009. Due to lower cost and a high efficacy of Spinosad it is likely farmers would
prefer this plant protection product. Considering resistance of the Colorado potato beetle it is
recommended to change the insecticides every year.

253-256

5.00 €

 

Update on life cycle of entomopathogenic nematodes
Ralf-Udo Ehlers

Extended abstract

259-260

0.00 €

 

Aiming to eradicate small hive beetle Aethina tumida using entomopathogenic nematodes
Andrew G. S. Cuthbertson, James J. Mathers, Lisa F. Blackburn, Gay Marris, Mike A. Brown, Giles E. Budge

Abstract: The small hive beetle (Aethina tumida) is an endemic parasitic pest and scavenger of
colonies of social bees indigenous to sub-Saharan Africa. In this region the beetles rarely inflict
severe damage on strong colonies since the bees have developed strategies to combat them.
However, A. tumida has since ‘escaped’ from its native home and has recently invaded areas such
as North America and Australia where its economic impact on the apiculture industry has been
significant. Commercially available entomopathogenic nematodes were screened for their
potential to control beetle larvae. The nematodes Steinernema kraussei and S. carpocapsae
provided excellent control with 100% mortality of larvae being obtained. Delayed applications of
the nematodes following larvae entering sand to pupate also provided excellent control for up to
3 weeks. The information gained supports the development of contingency plans to deal with
A. tumida should it occur in the UK or Europe.

261-265

5.00 €

 

The development of mollusc-parasitic nematode Phasmarhabditis hermaphrodita (Nematoda: Rhabditidae) in different substrates
Jiří Nermuť, Vladimír Půža

Abstract: The effect of different growing substrates on the development of the facultative slug
parasite Phasmarhabditis hermaphrodita has been studied in a series of laboratory experiments.
Wild, laboratory and Nemaslug strains of P. hermaphrodita were reared in agar plates on
homogenized pig kidney, the homogenized bodies of Deroceras reticulatum, Arion lusitanicus,
and Galleria mellonella, the faeces of D. reticulatum and A. lusitanicus, or leaf compost.
Development time, yield, lipid reserves, and the body length of females and dauer larvae were
assessed. All P. hermaphrodita strains were able to grow and reproduce on all tested substrates.
However, yields were markedly higher on animal substrates. Lipid content and body size varied
across the substrates, however, even plant tissue produced normal sized individuals with normal
lipid content. It thus seems that the quality of the substrate is expressed mainly in yield. High and
less variable yields and faster development of the wild and Nemaslug strains, in comparison with
the laboratory strain, were probably due to different bacterial associates. The dramatic differences
in yields on animal substrates, in comparison to those on plant tissue, illustrate the evolutionary
advantage of the association of nematodes with invertebrates.

267-270

5.00 €

 

New nematodes associated to Rhynchophorus ferrugineus (Coleoptera: Curculionidae): preliminary description
Monica Oreste, Francesca De Luca, Elena Fanelli, Alberto Troccoli, Eustachio Tarasco

Abstract only

271

0.00 €

 

The role of bacterial symbionts in the competition of entomopathogenic nematode species
Vladimír Půža, Jiří Nermuť, Zdeněk Mráček

Abstract: Competition between entomopathogenic nematode (EPN) species is still a largely neglected topic. Previous research has shown that in the competition within one insect host, nematode Steinernema affine strongly dominates over S. kraussei and suggested a possible role of symbiotic bacteria in the competition. In present study, S. affine and S. kraussei and their symbionts were reared in different combinations on Wouts agar plates, and nematode development was observed. Resulting progeny from these combinations was harvested and body size and lipid content of infective juveniles (IJs) were assessed. S. affine was able to develop, mature and produce viable progeny on the symbiont of S. kraussei. Interestingly, there was no difference in the duration of the cycle or reproduction potential, IJ size and lipid content between S. affine reared on their own symbiont and symbiont of S. kraussei. On the other hand, S. kraussei developed and reproduced well only on its own symbiont. These experiments explained the previously observed dominance of S. affine over S. kraussei. Research with more EPN species is planned to further clarify the topic.

273-276

5.00 €

 

Research and development for a nematode-based biological control solution for western corn rootworm in maize
Stefan Toepfer, Ulrich Kuhlman

Abstract: 10 years of joint efforts in research and development have led to a nematode-based
biological control solution for one of the most destructive maize pests, the western corn
rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae).

277-282

5.00 €

 

Development of a method to establish entomopathogenic nematodes (EPN) in arable soils by using farm-suitable field equipment
Wolfgang Büchs

Abstract only

285

0.00 €

 

Biosafety analysis of the Bacillus pumilus 15.1 strain through a Caenorhabditis elegans pathogenicity assay
Juan F. Caña Roca, Diana C. García, Juan I. Vilchez-Morillas, Maximino Manzanera, Tania Domínguez, Antonio Osuna, Susana Vílchez

Abstract: Using a Caenorhabditis elegans pathogenicity assay we evaluated the biosafety of the B. pumilus 15.1 strain, a recently isolated bacteria active against larvae of the Mediterranean fruit fly Ceratitis capitata. In the study we evaluated the toxicity of this strain toward the nematode together with other B. pumilus strains and compared its toxicity with a non pathogenic strain (Escherichia coli OP50) and a pathogenic one (Burkholderia cepacia). After this study, we concluded that B. pumilus 15.1 is a safe strain and could not represent a problem to be used as a biological control agent.

287-290

5.00 €

 

The indigenous entomopathogenic nematode searching results at different agrocenosis of Georgia
M. Chubinishvili, Ts. Chkhubianishvili, M. Kakhadze, I. Malania, I. Rijamadze

Abstract only

291

0.00 €

 

Field evaluation of entomopathogenic nematodes for controlling fall webworm Hyphantria cunea (Lepidoptera: Arctiidae) in West Georgia
Oleg Gorgadze, Manana Lortkipanidze, Patrick Tailliez, Medea Burjanadze, Madona Kuchava

Abstract: The present work deals with results of application of entomopathogenic nematodes of the genus Steinernema (S. carpocapsae, S. thesami and Steinernema sp.) against the harmful pest of the forest and agricultural crops Hyphantria cunea (Lepidoptera: Arctiidae) distributed in Georgia. Field experiments were carried out in August of 2012 on private plots of Guria region of the West Georgia in hazelnut plantations diseased with pest’s larvae. A high percentage of mortality ranging from 93.6% to 98.3% was observed in all experiments as a result of entomopathogenic nematode application. Among the species used, the efficiency of a new Steinernema species was specially noticed. High efficiency of the treatment was also promoted by optimum climatic conditions (Temperature = 28 °C and hygrometry = 99%).

293-296

5.00 €

 

Feeding activity and survival of slug Arion lusitanicus (Gastropoda: Arionidae) exposed to the rhabditid nematode, Phasmarhabditis hermaphrodita (Nematoda: Rhabditidae)
Dinka Grubišić, Tina Hamel, Tanja Gotlin Čuljak, Ana Loparić, Mirjana Brmež

Abstract: Slugs are important pests of cultivated plants in Croatia. In many sites slug species Arion lusitanicus (Mabille, 1868) (Gastropoda: Arionidae) has become the most frequent species, which is very hard to control by chemical molluscicides. Since 1996 a biological molluscicide based on nematode Phasmarhabditis hermaphrodita (Schneider, 1859) (Nematoda: Rhabditidae) has been formulated as an effective product for slug control. In order to establish feeding activity and survival of adult A. lusitanicus specimens exposed to parasitic nematode P. hermaphrodita and to compare its efficiency to efficiency of chemical molluscicides, a laboratory experiment was performed. Adult specimens of A. lusitanicus (feeded on lettuce leaves in flower pots) were exposed to P. hermaphrodita, metaldehyde and methiocarb treatments. Food consumption of slug specimens was measured daily. Survival of slugs was observed to their death, up to 30th day. In the first week of investigation, chemical molluscicide treatments were found to differ significantly from the biological product and control. At the treatments treated by nematodes, daily leaf area consumption was also reduced and was significantly different from the control treatment. Food consumption was reduced on both chemical and biological treatments but the most of adult speciments of A. lusitanicus survived and continued to feed. To the end of the second week of investigation, food consumption decreased on all treatments and was mainly uniform with no significant differences between treatments. During the experiment, the slugs were dying within the period of 3 to 30 d at the treatments treated by nematode P. hermaphrodita or in the period of 9 to 24 d at the treatments treated by metaldehyde and methiocarb. Because the tolerance level to slug damages in lettuce market is effectively zero, these results indicate a failure of biological product based on P. hermaphrodita in control of adult specimens of A. lusitanicus as well as a failure of chemical molluscicides. These data point at a great need for integrated control of slug damage which must include cultural and different alternative control measures, not only chemical control.

297-300

5.00 €

 

New insights to insect response to the infection by nematobacterial complex
Pavel Hyršl, Pavel Dobeš, Badrul Arefin, Lucie Kučerová, Robert Markus, Zhi Wang, Michal Žurovec, Ulrich Theopold

Abstract only

301

0.00 €

 

Compatibility of five different entomopathogenic nematode (Nematoda: Rhabditida) species with registered insecticides and fungicides under laboratory conditions
Žiga Laznik, Stanislav Trdan

Abstract: To increase our knowledge on the susceptibility of entomopathogenic nematode (EPN) species to agrochemicals, the compatibility of the infective juveniles (IJs) of the Steinernema feltiae, S. carpocapsae, S. kraussei, Heterorhabditis bacteriophora and H. downesi with 6 chemical, one plant-based and one bio-insecticide, and 13 synthetic organic and two inorganic fungicides were investigated under laboratory conditions. The effect of direct IJ exposure to insecticides for 6 and 24 hours was tested in Petri dishes at 15, 20 and 25 °C. In our experiment we determined the best compatibility of S. feltiae with active ingridients azoxystrobin, azadirachtin, Bacillus thuringiensis var. kurstaki and imidacloprid. The present study showed that S. carpocapsae and S. kraussei are sensitive to all tested insecticides, while H. bacteriophora is sensitive only to abamectin and lufenuron. Nematode H. downesi significantly suffered the highest mortality when infective juveniles were mixed with active ingredients (a. i.) tebuconazole, spiroxamine, and triadimenol. Based on our research, we conclude that compatibility is not only a species-specific but also a strain-specific characteristic.

303-308

5.00 €

 

Susceptibility of Phytodecta fornicata (Coleoptera: Chrysomelidae) to Heterorhabditis bacteriophora
Ivana Majić, Emilija Raspudić, Marija Ivezić, Mirjana Brmež, Ankica Sarajlić, Andrea Mirković

Abstract: The infectivity of Heterorhabditis bacteriophora for adult stage of Phytodecta fornicata was evaluated in the laboratory. Two different nematode concentrations (1000 and 2000 infective juveniles (IJs) per adult) were tested at temperatures of 22 °C and 30 °C. Mortality of insects was checked at 3 d post infection (p.i.); emergence of IJs from cadavers was noted at 8 and 11 d p.i.. For the first time, the experiment demonstrated that H. bacteriophora applied in rate of 1000 IJs per beetle is causing 100% mortality of P. fornicata, and that IJs developed inside adult insect cadavers. The effects of temperature and concentration of nematode preparation were of less importance. The harvest of IJs from cadavers was obtained but with low population level. The time of emergence of IJs was temperature dependent. In order to develop cost effective and sustainable control measure, further mortality studies on individuals, with other species and strains, and assays with more realistic environments are needed.

309-312

5.00 €

 

The susceptibility of mulberry moth to infection by entomopathogenic nematodes, Heterorhabditis bacteriophora and Steinernema carpocapsae
Nona Mikaia

Abstract: The mulberry moth, Glyphodes pyloalis, is considered as an urban pest and therefore the application of environmentally safe means for mulberry trees protection is recommended. Among the entomopathogenic nematode (EPN) species Heterorhabditis bacteriophora and Steinernema carpocapsae are important as a biological control agents. The susceptibility of G. pyloalis to infection by H. bacteriophora and S. carpocapsae infective juveniles (IJ) was tested under laboratory conditions. Individuals of IV instar larvae were collected from mulberry trees in Georgia, Tbilisi (village Digomi). Nematode suspensions at a concentration of 1500 IJs/ml were used for treatment of mulberry leaves. After 72 h, the mortality of G. pyloalis caused by H. bacteriophora was 54%, whereas S. carpocapsae caused 76% mortality. The results suggest that nematode suspensions of H. bacteriophora and S. carpocapsae can be used to control G. pyloalis in urban plots.

313-316

5.00 €

 

Attract and kill against western corn rootworm larvae with entomopathogenic nematodes
Mario Schumann, Felicitas Kaemena, Anant Patel, Stefan Vidal

Abstract only

317-318

0.00 €

 

Cloning strategy for recovering phage-displayed Cry1Aa13 mutants from phages with affinity towards proteins present in the gut of Ceratitis capitata
Tania Domínguez, Juan F. Caña Roca, Diana C. García, Antonio Osuna, Susana Vílchez

Abstract: Using the phage display technique, a pool of phages from a library of bacteriophages expressing Cry1Aa13 toxins with modified loops 2 at the domain II was selected that showed affinity toward proteins present in the guts of the Mediterranean fruit fly, Ceratitis capitata. The sequences of the hypervariable regions of the in vivo selected phages were analysed and an almost identical sequence was obtained in all of the selected phages. Those phages bearing toxins different from the wild type toxin at the loop 2 were selected in order to recover the Cry1Aa13 mutant toxins. Here we describe the cloning strategy designed and used to clone the toxins from the phage genome in order to be expressed.

321-325

5.00 €

 

Efficacy evaluation of different Bacillus thuringiensis sv kurstaki strain EG2348 formulations against Malacosoma neustrium (Lepidoptera: Lasiocampidae)
Luca Ruiu, Achille Loi, Giovanni Falchi, Edith Ladurner, Andrea Braggio, Pietro Luciano

Abstract: Cork oak forest protection and management require continuous monitoring of defoliator moth species. Among these, the European tent caterpillar, Malacosoma neustrium L., can cause widespread and extensive defoliation of host plants, and the implementation of appropriate management programs becomes necessary. Sustainable control methods may include the use of entomopathogenic microrganisms, such as Bacillus thuringiensis serovar kurstaki (Btk). However, the formulation of the microbial control agents can be a key factor for the success of application programs. The results of an efficacy trial with different formulations of Btk strain EG 2348 against larvae of M. neustrium conducted in a cork oak forest in Sardinia (Italy) are reported. In the trial, a commercial and an experimental formulation of Btk strain EG 2348 were tested (henceforth Rapax® and Rapax Experimental) in comparison to two Btk-based reference products (Foray 48B® and Delfin®). Both formulations of Btk strain EG 2348 proved to be effective in controlling the pest.

327-330

5.00 €

 

Development of a new bio-insecticide for controlling lepidopteran pests
Kazım Sezen, Remziye Nalçacioğlu, Ismail Demir, Hüseyin Tepe, Islam Yildiz, Zihni Demirbağ

Abstract only

331

0.00 €

 

Bioluminescence determination of antibacterial activity of Bombyx mori and Galleria mellonella haemolymph
Libor Vojtek, Pavel Dobes, Ender Buyukguzel, Pavel Hyrsl

Abstract: We describe an antibacterial assay based on bioluminescence of two Gram negative bacteria Photorhabdus luminescens and transformed Escherichia coli, which can be used for a real-time measurement of antibacterial activity. We observed a significant dose-dependent decrease of bioluminescence using both bacterial species during one hour after exposure to Bombyx mori or Galleria mellonella haemolymph. The humoral origin of the antibacterial activity observed in whole haemolymph was confirmed in haemolymph plasma without haemocytes. Antibacterial activity operating against Gram negative bacteria was measured in unaffected insect larvae as well as after septic injury; increased antibacterial activity in haemolymph was detected in the latter case which confirms inducibility of antimicrobial agents. This method can be widely used for determination of antibacterial activity in insects and supposedly in other invertebrates.

333-338

5.00 €

 

Intramolecular cleavage at the loop between α3-helix and α4-helix is critical for cytotoxic activity of Cry8Da
Takuya Yamaguchi, Hisanori Bando, Shin-ichiro Asano

Abstract: Cry8Da from Bacillus thuringiensis galleriae SDS-502 has the toxicity against both larvae and adult P. japonica. Cry8Da is processed into three fragments (64 kDa, 54 kDa and 8 kDa) by gut juice of P. japonica. Fragments of 54 kDa and 8 kDa are derived from the cleavage of 64 kDa fragment at the loop between α3-helix and α4-helix in Domain I. Binding assays showed that the 54 kDa fragment bound to both larvae and adult P. japonica brush-border membrane vesicles while the 64 kDa and 8 kDa fragments did not. We constructed a protease-resistant mutant, 8Da-R163A, in which R163 on the loop was changed to A163. To directly investigate whether intramolecular cleavage is critical for insecticidal activity of Cry8Da, we performed cytotoxic assays against midgut epithelial cells (MECs) prepared from adult P. japonica using purified uncleaved (64 kDa) and intramolecular cleaved (mixture of 54 kDa and 8 kDa) Cry8Da toxin. Cytotoxic assay showed MECs were destroyed by only intramolecular cleaved Cry8Da toxin. Intramolecular cleaved Cry8Da toxin also formed oligomeric structure after incubation with MGCs. These results strongly support our idea that the cleavage at the loop between α3-helix and α4-helix is critical for toxicity of Cry8Da.

339-342

5.00 €

 

Electron microscope and genetic analysis of an intracellular bacterium associated with the common rough woodlouse, Porcellio scaber (Isopoda, Porcellionidae)
Regina G. Kleespies, Andreas Leclerque

Abstract: The common rough woodlouse, Porcellio scaber, is a common and widespread isopod species of Western and Northern Europe. A previously unknown intracellular bacterium has been identified in a diseased Porcellio larva. Microscopic studies revealed the subcellular structures characteristic of infection by Rickettsiella-like bacteria. Molecular phylogenetic analysis based on the 16S ribosomal RNA encoding rrs gene demonstrated that the woodlouse pathogen belongs to the taxonomic genus Rickettsiella (Gammaproteobacteria; Legionellales). Moreover, genetic analysis makes it likely that this new pathotype should be considered a member of the “Rickettsiella armadillidii complex”, i.e. a group of Rickettsiella bacteria found mainly in terrestrial isopods. R. armadillidii is currently placed in synonymy with the nomenclatural type species, Rickettsiella popilliae. The present study does not lend support to this synonymization.

343-347

5.00 €

 

Impact of various oilseed rape productions on biological potential of endogaeic active ground beetles (Coleoptera: Carabidae)
Ivan Juran, Tanja Gotlin Čuljak, Wolfgang Büchs, Dinka Grubišić, Ivan Sivčev

Abstract only

351

0.00 €

 

Ground beetles (Coleoptera: Carabidae) in sugar beet fields as the base for conservation biological control
Tomislav Kos, Renata Bažok, Zrinka Drmić, Željka Graša

Abstract: The fauna and abundance of ground beetles (Coleoptera, Carabidae) in arable crops can be an indicator of influence of different agricultural measures on biodiversity. The aim of our study was to determine ground beetle fauna abundance and frequency in two fields with different herbicide and insecticide application practice, and to determine differences in total number of species and individuals, collected with two capturing methods. The study was conducted in 2012 in the eastern part of Croatia (County of Vukovar-Srijem). Beetles were collected in a period of twenty-one weeks (April- September) by setting four modified pitfall traps aimed to collect above ground fauna and four probes (WB PROBE II® Trap, Trece inc.) aimed to collect endogeic fauna in each field. Nine different species and eight genera were identified in the study. Most abundant were Pseudoophonus rufipes (De Geer 1774) and Bembidion sp. (Latreille 1802). Both are classified as eudominant. The most frequent species was P. rufipes classified as constant (71.42%) and the most frequent genus was Bembidion sp. (38.04%) classified as accessory. There was no significant difference between fields among total number of established species and/or genus no matter if they were captured by pitfall trap or probe. Significantly more individuals were captured in pitfall traps on the field No. 1 (33.3) than on the field No. 2 (8.8), respectively. Opposite, significantly fewer individuals were captured with probe on the Field No. 1 (0.5) than on the field No. 2 (6.6), respectively.

353-357

5.00 €

 

Impact of Entomophaga maimaiga on gypsy moth populations in Bulgaria
Plamen Mirchev, Andreas Linde, Daniela Pilarska, Plamen Pilarski, Margarita Georgieva, Georgi Georgiev

Abstract: The entomopathogenic fungus Entomophaga maimaiga Humber, Shimazu and Soper (Entomophtorales) (Entomophtoraceae) was introduced into three populations of gypsy moth (Lymantria dispar L., Lepidoptera: Erebidae) in Bulgaria in 1999. After the first strong epizootics in 2005, the species was introduced in six outbreak populations of gypsy moth in different regions of the country from 2008 to 2011. Due to the resulting fungal epizootics, the calamities of the pest in Bulgaria were totally suppressed. The pathogen increased its impact by a natural range extension and it is now present in nearly all regions of the country in which L. dispar occurs.

359-363

5.00 €

 
Add this product to your basket