IOBC-WPRS Bulletin Vol. 111, 2015 Working Group “Integrated Protection of Stored Products”. Proceedings of the Meeting at Zagreb (Croatia), June 28 – July 1, 2015. Edited by Pasquale Trematerra and Darka Hamel. ISBN 978-92-9067-294-4 [XLIV + 530 pp.]

Contents Committees ........................................................................................................................... I Foreword by the local organizer .......................................................................................... III Preface by the convenor ...................................................................................................... V Group photo ...................................................................................................................... VII List of authors of oral presentations .................................................................................... IX List of authors of posters ................................................................................................... XII List of authors .................................................................................................................... XV Contents ....................................................................................................................... XXXV

Session 1. Physical, chemical and other techniques for stored product pest control

Chemical, physical and organic hermetic storage technology

for stored-product protection in African countries Keynote Speaker: Daniel Obeng-Ofori ..................................................................... 3-27 Abstract: A myriad of beetle and moth pests associated with durable grains and processed food

cause considerable quantitative and qualitative losses throughout the world. Currently, there is

lack of reliable and verifiable data on postharvest losses in most African countries. Many tools

are available for protecting stored food against pest infestation including the use of contact

insecticides and fumigants, botanicals, inert dusts, irradiation, biological control agents as well as

appropriate modified atmospheres through metal silo and hermetic storage technology. This

paper focuses on the advances in stored-product protection in sub-Sahara Africa with emphasis

on chemical, inert dusts and organic hermetic storage technology with the view to identifying

critical challenges and information gaps that need further research. Although pest management

strategies are changing to meet consumer’s demand for food free of chemical residues, address

concerns about safety of insecticides to humans, delay insecticide resistance development in

insects and comply with stricter pesticide regulations, the use of synthetic contact insecticides

will continue to be a major component of stored product pest management programmes. Selective

use of contact insecticides requires a thorough understanding and evaluation of risks, costs and

benefits. Inert dusts and organic hermetic storage technology may be a safe, cost-effective and

environmentally friendly method of grain preservation against pest infestation among low-

resource poor farmers in developing countries who store small amounts of grains. It must be

emphasized that for sustainable stored product protection, a combination of the various control

options in an integrated and compatible manner is imperative. Integrated pest management (IPM)

as a knowledge-based system, is rapidly providing a framework to reduce dependence on

synthetic chemical pesticides. A major research priority is well-designed on-farm trials to

validate the efficacy of locally available inert dusts for stored-product protection using standard

procedures and formulations that can be transferred to other communities. The current knowledge

of organic hermetic storage technology, its availability and adoption is limited. Appropriate

knowledge transfer techniques needs to be developed and disseminated widely to encourage the

uptake of the technology for grain protection in Africa.

Toxicity of Turkish diatomaceous earth deposits

against some stored-grain insects on wheat Ali A. Işikber, Özgür Sağlam, Mehmet Kubilay Er, Hasan Tunaz, İnanç Ş. Doganay, Recep Sen .................................................................................... 29 Abstract only

Enhancing the insecticidal efficacy of inert dusts against stored food insect pest

by the combined action with essential oils Hind Houria Bougherra-Nehaoua, Stefano Bedini, Francesca Cosci, Guido Flamini, Kamel Belhamel, Barbara Conti ....................................................... 31-38 Abstract: Inert dusts (IDs), as Kaolin (K) and diatomaceous earth (D) powders, and aromatic

plant essential oils (EOs) are among the most appealing alternatives to traditional chemical

insecticides, against stored-products and field crops pests. IDs and EOs are environmentally

friendly and very low toxic to mammalian. In addition, it has been proven that insects are not

able to develop resistance to IDs. Recent studies indicate that, when used in combination,

aromatic plant EOs may enhance the insecticidal efficacy of IDs. Such effect could be due both

to the physical action of IDs on the cuticle and the supposed neurotoxic effects of EOs.

The aim of the present study was to evaluate the efficacy of the two aromatic plants Pistacia

lentiscus and Foeniculum vulgare EOs, of K and D powders and of the IDs-EOs mix as grain

protectants against the maize weevil Sitophilus zeamais (Motschulsky) (Curculionidae). The EOs

were tested at 10, 40, 80 and, 100 ppm, IDs at 25, 50 and 100 ppm while the EOs + IDs at 20, 40

and 60 ppm in 330 ml pots containing 150 g of maize grains and 10 unsexed insects. Results

were recorded after three, six and nine days of treatment. Orthogonal contrasts analysis shows

that the action of IDs was higher when used in combination with the EOs (P < 0.05). These

results indicate that the combined use of IDs and EOs could effectively enhance the insecticidal

efficacy of IDs reducing the costs of the management of stored grains.

Short exposures to phosphine in combination with low pressure: does it work?

Basilis Sotiroudas, Christos I. Rumbos, Maria Sakka, Christos G. Athanassiou ............ 39 Abstract only

Phosphine resistance in eggs of Tribolium castaneum and Plodia interpunctella

from almond storage facilities in the Central Valley of California Sandipa Gautam, George P. Opit ........................................................................... 41-49 Abstract: Phosphine (PH3) is heavily relied upon for stored-product pest management

worldwide. Several studies have revealed high frequencies of PH3 resistance in stored-product

insects in many countries including the United States. These studies focused on adults, usually

the most fumigant-susceptible life stage. In this study, we established discriminating doses of

PH3 for Tribolium castaneum (red flour beetle, RFB) and Plodia interpunctella (Indian meal

moth, IMM) eggs using laboratory susceptible strains for detection of resistance in eggs, the most

fumigant-tolerant life stage. The discriminating doses for RFB and IMM eggs were established

based on 3-d fumigation and were 73.6 and 109.8 ppm, respectively. Discriminating dose tests

showed that eggs of four out of 11 RFB field populations collected from almond storage facilities

in California had resistance frequencies (percentage survival) that ranged from 54-100%. All

IMM populations tested were resistant to PH3 and resistance frequencies ranged from 4-20%.

Based on 3-d fumigation, dose-response tests and probit analyses determined that LC99

(concentration required to kill 99% of the eggs) values for susceptible and the most resistant RFB

eggs were 50.8 and 653.9 ppm, respectively. Resistance frequencies and levels of resistance in

different life stages of RFB and IMM and their implications for resistance management are

discussed.

Persistence and efficacy of diatomaceous earth, imidacloprid and Beauveria bassiana

(Balsamo) Vuillemin against three Coleopteran and psocid species of stored grains Waqas Wakil, Thomas Schmitt..................................................................................... 51 Abstract only

Efficacy of six dry formulations of insecticides

against five major stored-product insect species Bhadriraju Subramanyam, Xinyi E., Spencer Diveley ............................................. 53-59 Abstract: Six newly developed dry insecticide formulations containing pirimiphos-methyl and

deltamethrin were loaded onto licocene, a waxy material used as a filler, to decrease the

recommended application dosages of these insecticides. Out of the six formulations, three were

formulated with diatomaceous earth as an inert ingredient and three without. Formulations A, B,

E, and F containing pirimiphos-methyl (0.5 ppm) and deltamethrin (0.12 or 0.24 ppm) were

applied to corn, whereas formulations C and D containing only deltamethrin (0.24 ppm) were

applied to both corn and wheat. Adults of the red flour beetle, sawtoothed grain beetle, rice

weevil, lesser grain borer, and eggs of Indian meal moth were exposed to untreated and treated

grain at 28 °C and 65% RH. All adults of the beetle species died after a 7- and 14-day exposure

and no progeny were produced on insecticide-treated grain. Live larvae and adults were observed

on untreated grain and no larvae or adults were found in all formulations except formulations D

and E. All formulations performed well against the beetle species and the Indian meal moth, and

have potential as candidate grain protectants.

Establishing discriminating doses of phosphine for adults of three psocid

(Psocoptera: Liposcelididae) species Charles E. Konemann, George P. Opit, Sandipa Gautam ...................................... 61-65 Abstract: Psocids are now recognized as stored-product pests of substance worldwide. Lack of

information exists on resistance of psocids to phosphine (PH3) in the USA. In this study we

determined discriminating doses of PH3 for adults of three psocid (Psocoptera: Liposcelididae)

species, namely, Liposcelis bostrychophila Badonnel, L. paeta Pearman, and L. decolor

(Pearman). Protocols used were modified from the FAO Method No. 16, to determine lethal

doses that caused 99% mortality and their 95% confidence intervals for the laboratory susceptible

strain of each species. Dose-response studies were conducted by exposing adults of all species to

different concentrations of PH3 for 20 hours. Mortality was assessed 24 hours after exposure.

Discriminating doses for L. bostrychophila, L. paeta, and L. decolor adults were 46.7, 122.2, and

187.8 ppm, respectively. These data show that the three Liposcelis species responded differently

to PH3. The discriminating doses from this study will be used in future research to determine

resistance frequencies of field populations of these species.

Large-scale applications of nitrogen against stored-product insects

in commercial facilities Christos G. Athanassiou, Antonia Chiou, Christos I. Rumbos, Maria Sakka, Basilis Sotiroudas, Vaios Karathanos .......................................................................... 67 Abstract only

Survival and development of stored product insects' eggs in baby food

exposed to low oxygen Hagit Navarro, Shlomo Navarro ............................................................................. 69-76 Abstract: Food industry has been increasingly using modified atmospheres for the protection of

the packaged commodities from potential insect contamination in the product. This is especially

so for foods which are sensitive to insect contamination like baby foods where packaging is

carried out under nitrogen (N2). Canned packages were sealed and treated using N2 to achieve an

initial concentration of 1% oxygen (O2). This concentration was shown to increase to 2.5% due to

desorption of O2 from the commodity. The increase in customer complaints on presence of stored

product insects in baby food formula led the manufacturer to the questions (1) whether adult

insects can lay eggs in milk powder formula after its packaging at 1% O2; and (2) whether the

eggs can develop at such low oxygen concentration. To answer those questions two sets of trials

were made; (a) testing the egg stage of Lasioderma serricorne, Plodia interpunctella and

Oryzaephilus surinamensis exposed to 2.5% and 6% O2 on baby food, containing mainly milk

powder, that had equilibrium relative humidity of 46.6%, and (b) testing the survival of the adults

of same insects and their ability to lay eggs at 2.5% and 6% O2. For the test purpose, adults of

L. serricorne, P. interpunctella and O. surinamensis were reared at ambient air at 29 ± 1 °C and

65% ± 5 RH. Results show that eggs of the tested three species continued to develop after they

were exposed to 6% O2. However, larvae could not survive the same oxygen concentration and

no pupae neither adult's stage were observed in the treated media. In an atmosphere containing

2.5% O2, eggs of L. serricorne and P. interpunctella failed to develop, but only O. surinamensis

eggs hatched. The ability of adults of P. interpunctella to lay eggs was much higher than

L. serricorne, in the same atmosphere. Whereas, adults of O. surinamensis failed to lay eggs in

both O2 concentrations tested.

The entomocidal and growth inhibitory impact of three formulations

of diatomaceous earth in combination with essential oils of Allium sativum and Azadirachta indica against Rhyzopertha dominica (F.) Muhammad Sagheer, Mansoor-Ul-Hasan, Muhammad Sajid Qureshi, Sidra Tul Muntaha, Sehrish Kanwal, Qurban Ali, Kazam Ali, Fawad Zafar Ahmad Khan .......................................................................................... 77 Abstract only

Insecticidal effect of various insecticides for the control of the khapra beetle,

Trogoderma granarium Everts (Col.: Dermestidae) on concrete Christos G. Athanassiou, Nickolas G. Kavallieratos, Maria C. Boukouvala, Marios Mavroforos, Dimitrios C. Kontodimas, Myrto S. Barda ..................................... 78 Abstract only

Heat treatment in food industries: efficacy for the insects and prospect

for a microbiological reduction Paolo Guerra .......................................................................................................... 79-84 Abstract: This paper presents the results achieved at the end of a mill treatment carried out in

2014. Temperature and humidity were regularly monitored in the treated premises; the

effectiveness of the treatment was checked by means of biological tests. The data collected show

that, keeping the average temperature of 52.8 °C, with no side effects for machineries and milling

structures, the mortality rate reached 100% for all the stages of Ephestia kuehniella, Sitophilus

granarius and Tribolium confusum. Some microbial analysis have been carried out before and

after the treatment, showing a decrease up to 60% in the microbial load e.g. moulds and airborne

bacteria.

Optimising the delivery of pesticide to grain using Entostat® to reduce the amount

required for effective control Freya Scoates, Adam Nunn, Charlotte Elston ........................................................ 85-94 Abstract: There is significant pressure to reduce the use of chemical pesticides in stored grains

and other dried commodities for operator safety and because residues can lead to consumer

rejection. However, growers are keen to keep pesticides as one of a suite of tools under an IPM

approach. Optimising the delivery of the pesticide can reduce the amount required for effective

control. Entostat® is an electrostatic micro-powder formulated using blends of waxes and

polymers, which can have an active ingredient dispersed within its matrix. It adheres to insect

cuticles and grain kernels and thus can be used to effectively target storage pest species,

increasing the exposure of the insect to the pesticide and evenly distributing in grain during

admixture. Deltamethrin and pirimiphos-methyl were formulated in Entostat, without the

synergist PBO, at rates ≤ half that of a conventional deltamethrin-based formulation. In a series

of studies control of four significant grain pests: Sitophilus granarius (L.), Oryzaephilus

surinamensis (L.), Rhyzopertha dominica (F.) and Tribolium castaneum (Herbst), was achieved

equivalent to the conventional formulation. We demonstrated that by improving the delivery,

using a carrier such as Entostat, it is possible to apply chemical pesticides at much lower

concentrations than are currently used, while achieving equivalent levels of control. This

indicates that there is significant potential for chemical pesticides to continue as an important

component in storage IPM.

Application of the fumigant methyl isothiocyanate to wheat: fumigant toxicity,

sorption dynamics, germination effects and residues after storage Özgür Sağlam, Michael J. Aikins, Thomas W. Phillips ................................................. 95 Abstract only

Effectiveness of Profume® gas fumigant (sulfuryl fluoride) for the control

of phosphine-resistant grain insects infesting stored wheat George P. Opit, Ellen Thoms, Thomas W. Phillips ............................................... 97-103 Abstract: Highly phosphine-resistant populations of Rhyzopertha dominica (F.) (Coleoptera:

Bostrichidae) and Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) have been found in

Oklahoma grain storage facilities. The phosphine-resistant R. dominica and T. castaneum

populations found are up to 1,519 and 119 times, respectively, more resistant than their

susceptible counterparts. These findings call for development of phosphine resistance

management strategies to ensure continued effective use of phosphine. Eliminating highly

phosphine-resistant insects using an alternative fumigant such as Profume® gas (sulfuryl fluoride

or SF) can be a component of phosphine resistance management. Therefore, we conducted an

experiment in eight 13.6-MT steel bins to determine the efficacy of SF to control phosphine-

resistant R. dominica and T. castaneum infesting stored wheat. Based on our data, SF was highly

effective in controlling all stages of phosphine resistant R. dominica and T. castaneum. The

effectiveness of SF to control phosphine-resistant insects and implications of this for resistance

management are discussed.

Novel insecticide formulations using Entostat powder technology:

effects on stored product beetles and on the commodity Christos G. Athanassiou, Thomas N. Vassilakos, Anna-Cristina Dutton, Nick Jessop, David Sherwood, Garry Pease, Andreja Brglez, Clare Storm, Stanislav Trdan ......................................................................................................... 105 Abstract only

The potential of Croatian diatomaceous earths as grain protectant

against three stored-product insects Anita Liška, Vlatka Rozman, Zlatko Korunić, Josip Halamić, Ines Galović, Pavo Lucić, Renata Baličević ............................................................................. 107-113 Abstract: The objective of this study was to determine the potential insecticide efficacy of

several Croatian diatomite samples (as inert dusts) against Sitophilus oryzae (Linnaeus),

Tribolium castaneum (Herbst) and Rhyzopertha dominica (Fabricius) adults and to compare their

efficacy with the efficacy of the USA standard diatomaceous earth (DE) Celatom Mn 51.

DE Mn 51 belongs to a group of DE with medium to increased efficacy for stored agricultural

products insects. Six Croatian samples from three different locations (MA-4, MR-10, MR-10B,

OP-4, OP-4A and PD-1) were selected and prepared for testing with fractions of particles

≤ 45 µm and applied at 500 mg/kg. Insect mortality was recorded after 7 and 14 days. The

highest lethal effect after 7 and 14 days showed Croatian sample MA-4 with 97% and 100%

mortality, respectively, for S. oryzae, 61.5% and 99% mortality, respectively, for T. castaneum

and 26.5% and 34%, respectively, for R. dominica. In all treatments the efficacy of MA-4 was in

the same range with the efficacy of the standard DE Mn 51. The other five tested samples had

significant lower efficacy regarding to MA-4, although after 14 days mortality was 85.7-98% for

S. oryzae, 5-47% for T. castaneum and 2-5% for R. dominica depending on a different DE

sample. These results show that Croatia has potential diatomite deposits with good and promising

efficiency for tested insects.

Immediate lethal dose of gamma radiation to Callosobruchus maculatus adults

Fabrício Ca. Reis, Marcos Roberto Potenza, Valter Arthur ........................................ 115 Abstract only

Effect of microwave on different stages of dates moth, Ephestia cautella (Walker)

(Lepidoptera: Pyralidae) in stored dates Mohammed Z. Khalaf, Falah H. Naher, Hussain F. Alrubeai .............................. 117-123 Abstract: Dates moth (fig moth), Ephestia cautella (Walker) is a major pest in storage and dates

on the market. Eggs, larvae, pupae and adults of date moth E. cautella were exposed to three

levels of microwave power (600, 800 and 1000 watts) at different exposure time (0, 6, 8, 10, 12,

14, 16, 18 and 20 seconds). The results showed that microwave had high ability to kill different

stages of date moth. The effect increased by increasing of microwave power and exposure time.

The percentage of egg mortality was 100% at 600 and 800 W, and 14 sec exposure time.

Similarly, eggs mortality was also high at 1000 W, but in less than 12 sec. The highest mortality

of larvae was 90% at 600 W and 96.7% at 800 W, when the exposure time was 10 sec The

mortality was 100% at 1000 W and exposure time was 10 sec Male and female pupal mortality

rate was 100% at 800 W, and 20 or 18 sec respectively. The range of lethal time of 50% and 95%

(LT50, LT95) at 600-1000 W were 5.01-9.9 and 15.7-34.2, 3.0-7.5 and 9.0-50.0 and 3.2-4.8 and

7.9-39.1 for egg, larvae and pupae respectively. The results showed that all treatments of adults

significantly affected longevity and emergence of adult male and female. The longevity of

emerged males and females did not exceeded 4.3 and 5.3 days, respectively at 600 W and 6 sec

whereas emerging of adults stopped at 800 W and 14 and 16 sec, for male and female pupae,

respectively. The results of this investigation indicated the feasibility of microwave technique

and could be used as alternative to methyl bromide in stored dates.

Effectiveness of wheat-applied contact insecticides against Sitophilus oryzae (L.)

(Coleoptera: Curculionidae) Marijana Pražić Golić, Petar Kljajić, Goran Andrić .............................................. 125-132 Abstract: The effectiveness of several contact insecticides: malathion, chlorpyrifos-methyl,

pirimiphos-methyl, deltamethrin+piperonyl butoxide (PBO) synergist, bifenthrin, thiamethoxam,

spinosad and abamectin against a laboratory population of Sitophilus oryzae in treated wheat, was

examined. Adult mortality was determined after 2, 7 and 14 days of exposure, while minimum

effective doses (MED) were determined based on F1 and F2 offspring production after 8 and 14

weeks, respectively. Chlorpyrifos-methyl (LD50 = 0.36 mg/kg) and pirimiphos-methyl

(LD50 = 0.43 mg/kg) were the most effective contact insecticides after 2 days of exposure, while

abamectin was the least effective (LD50 = 23.53 mg/kg). Chlorpyrifos-methyl (LD50 = 0.13 mg/kg)

had the highest effectiveness after 7 days of exposure, and malation the lowest

(LD50 = 1.26 mg/kg), while abamectin (LD50 = 0.07 mg/kg) was the most effective insecticide

after 14 days, and malathion the least (LD50 = 1.11 mg/kg). Chlorpyrifos-methyl and deltamethrin

had the lowest MEDs (> 0.5 and 0.6 mg/kg, respectively) in F1 generation, and deltamethrin and

bifenthrin (0.4 and 0.5 mg/kg, respectively) in F2 generation, while spinosad showed the highest

MED both in F1 and F2 with > 20 and 10 mg/kg, respectively.

The effect of diatomaceous earth treatment combined with 50 °C temperature on Sitophilus oryzae (L.) in wheat grain Marijana Pražić Golić, Petar Kljajić, Goran Andrić .............................................. 133-138 Abstract: Diatomaceous earth (DE) and extreme temperatures (high or low) used in mutually

independent treatments make effective alternatives to residual insecticides but many limiting

factors prevent their wider use. To minimize such negative effects, we have examined the

influence of a treatment with DE originating from Serbia in combination with 50 °C temperature

on a laboratory and a field (Novi Pazar) population of S. oryzae. Wheat grain was treated with

DE (0.25, 0.5 and 0.75 g/kg) before S. oryzae adults were added to grain in plastic cups and

placed in a temperature chamber set to 50 °C for periods of 65 and 75 min. After each exposure

period, the plastic cups were transferred to 24 °C in the laboratory, and lethal effects were

checked after 1, 2, 7 and 14 days, while offspring insects were counted after 8 weeks. For

comparison, the effects of DE on weevils at 24 °C, and effects of exposure to 50 °C temperature

were recorded in separate bioassays. Adults of both populations in plastic cups not exposed to

50 °C temperature reached mortality of > 95% after 14 days in wheat grain treated with 0.5 and

0.75 g/kg DE. Regarding both weevil populations exposed to 50 °C temperature for 65 min, total

mortality and 100% offspring reduction were found after 7 days of contact with 0.25-0.5 g/kg

DE. The results of the present study reveal that a combination of DE and 50 °C temperature may

be preferable to their independent use.

Toxicity of gaseous ozone at high concentrations against Plodia interpunctella

(Hübner) and Ephestia cautella (Walker) in in-shell hazelnuts Ali A. Işikber, Christos G. Athanassiou, K. Sinan Dayisoylu, M. Serdar Öztekin, Ahmet D. Duman, Melek Özdemir .............................................. 139 Abstract only

The efficacy of controlled atmosphere on Oryzaephilus surinamensis

under various temperature regimes Radek Aulicky, Jan Plachy, Vlastimil Kolar, Vaclav Stejskal ...................................... 140 Abstract only

Research on insecticidal efficacy of different inert dusts against the maize weevil

(Sitophilus zeamais Motschulsky, Coleoptera, Curculionidae) adults Stanislav Trdan, Aleksander Horvat, Tanja Bohinc ............................................. 141-145 Abstract: In the search for an effective and sustainable control method against the maize weevil

(Sitophilus zeamais Motschulsky), an important insect pest affecting stored grain, different inert

dusts were tested under laboratory conditions. We treated wheat grains with quartz sand, zeolites,

and diatomeaceous earth. Inert dusts of different origins were used, namely diatomaceous earth

from Slovenia and SilicoSec, quartz sands from two locations from Slovenia, and three different

zeolites (two types of natural zeolite from location in Slovenia, and synthetic zeolite Asorbio®).

Untreated winter wheat grains served as control treatment. The substances were tested at three

different temperatures (15, 20 and 25 °C) and two different relative humidity levels (55 and

75%). Mortality was measured 7th, 14

th and 21

st day after exposure. Inert dusts were applied at

two different concentrations, 450 and 900 ppm. The analysis of pooled results provoked

significantly the highest mortality of beetles in treatments with SilicoSec® (52.31 ± 2.07%), and

in treatment with one type of Slovenian zeolite (31.48 ± 1.42%). The lowest mortality was

recorded in treatments with quartz sands from both Slovenian locations, Moravče

(18.84 ± 1.31%), and Raka (9.12 ± 0.66%). Mortality of S. zeamais was significantly the highest

in treatments exposed to 25 °C (28.32 ± 1.16%), and in treatments exposed to higher

concentrations (900 ppm) of inert dusts (27.30 ± 0.87%). The use of diatomaceous earth is well

established in stored products pest management, however the knowledge on the efficacy of

zeolites is very week and offers a lot of opportunities for future researchers.

Study of synergetic impact of dust grain protectants combined with atmospheric modifications by controlling oxygen and ozone concentrations Dusan Losic .............................................................................................................. 147 Abstract only

Combination of diatomaceous earth formulations against larvae and adults

of Tribolium castaneum on wheat grains Waqas Wakil ............................................................................................................. 148 Abstract only

Insecticidal effect of the combined application of diatomaceous earth and zeolite

against three major stored-product insects in wheat Christos I. Rumbos, Maria Sakka, Nickolas G. Kavallieratos, Christos G. Athanassiou ..................................................................................... 149-156 Abstract: Diatomaceous earths and zeolites are both promising alternatives to chemical grain

protectants for the control of post-harvest infestations in dry, durable commodities. However,

there is no report on the combined application of diatomaceous earth and zeolite against stored-

product insects. In the present study, the insecticidal effect of eight diatomaceous earths and two

zeolites of different origins was evaluated against adults of Sitophilus oryzae (L.) (Coleoptera:

Curculionidae), Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae) and Oryzaephilus surinamensis (L.) (Coleoptera: Silvanidae) at two doses (250 and 1000 ppm) and

three exposure intervals (2, 7 and 14 d). After 2 d of exposure, mortality was low for all tested

species and doses, whereas after 7 d mortality ranged between 21.7 and 93.3 for S. oryzae, and

26.7 to 98.3 for O. surinamensis at 1000 ppm. Complete control (100%) was achieved only in the

case of S. oryzae and O. surinamensis after 14 d of exposure in wheat treated with the high dose. Tribolium confusum was the most tolerant to diatomaceous earth and zeolite among the species

tested. In a second series of bioassays, the three best performing diatomaceous earths and the

most effective zeolite were evaluated against the same beetle pests alone or in combinations at

250 and 500 ppm (total concentration of either one compound alone or both compounds). In all

cases, no significant synergistic effect was observed after the combined application of

diatomaceous earth and zeolite.

Modified atmospheres with high carbon dioxide in a new pneumatic chamber

for the control of stored product pests Pompeo Suma, Alessandra La Pergola, Salvatore Bella, Marco Amante, Pietro Ranno, Agatino Russo .................................................................................... 157 Abstract only

“Redrying” procedure in stored dried tobacco for control of Lasioderma serricorne

and Ephestia elutella Smaragdi Ch. Papadopoulou, Constantinos Chryssohoides, Constantinos Th. Buchelos ....................................................................................... 158 Abstract only

Efficacy of chlorine dioxide gas against five stored-product insect species

Bhadriraju Subramanyam, Xinyi E. ..................................................................... 159-168 Abstract: In this study, the efficacy of chlorine dioxide gas as a potential fumigant to control

stored-product insects was evaluated. Adults of five species of economically important stored-

product insects were exposed for varying time periods to four concentrations of chlorine dioxide

gas. Mortality was observed on days 1 through 5 after exposure to record any delayed mortality

effects. Phosphine susceptible laboratory strains and phosphine resistant field strains of the five

species were exposed to chlorine dioxide. The five species tested included the lesser grain borer,

red flour beetle, sawtoothed grain beetle, rice weevil, and maize weevil. Exposure to 0.54 g/m3

(200 ppm) of chlorine dioxide for 8 h at an average temperature of 18 °C and an average

humidity of 20% only resulted in less than 40% mortality of the five species. All exposed

sawtothed grain beetles died within a day when exposed for 6 h to a chlorine dioxide

concentration of 1.35 g/m3 (500 ppm). Red flour beetle, rice and maize weevils, and lesser grain

borer were more tolerant to this concentration, with mortality ranging between 25 and 80%. An

exposure to a chlorine dioxide concentration of 2.02 g/m3 (750 ppm) for 6 h produced trends in

mortality of all five species that was similar to mortality observed at 500 ppm. The mortality of

phosphine-susceptible and phosphine-resistant sawtoothed grain beetle, red flour beetle, and

lesser grain borer adults was 100% mortality when exposed for 7 h to a chlorine dioxide

concentration of 2.70 g/m3 (1000 ppm). Only the phosphine susceptible rice weevil showed

100% mortality at this concentration. Phosphine resistant rice weevil and maize weevil had 57

and 50% mortality when exposed for 7 h to a chlorine dioxide concentration of 2.70 g/m3. At this

concentration and exposure time, phosphine susceptible maize weevil showed 83% mortality one

day after exposure. The post-exposure mortality increased from day 1 to day 5 suggesting

delayed mortality effects after exposure to chlorine dioxide. Chlorine dioxide may be a potential

gas to control laboratory and field strains of the five species tested.

Session 2. Methods of pest prevention during storage, transportation and handling

of stored products Pest prevention during storage, transportation and handling of stored products

Keynote Speaker: Vaclav Stejskal ...................................................................... 171-176 Abstract: Cropping and storage systems are considered more resilient to pests when their

inherent properties reduce pest occurrence and pressure or limit yield losses in case of pest

attacks. Increased resilience of storage and food production systems includes an integration of

complementary preventive tactics and techniques into two defensive lines. First defensive line

(i.e. resilience of the store/production facility) represents prevention of entry to buildings.

Prevention of pest entry includes exclusion of active pest introductions (invasion or penetration)

and exclusion (quarantine) of passive introductions either via infested packages or with hidden

infestation of raw products (“Trojan-horse infestation”). Second defensive line (i.e. resilience of

commodity) represents prevention of pest multiplication in buildings and prevention of pest

invasion into a raw or processed commodities and multiplication inside them. The following

techniques and tactics can be employed to ensure internal security: “first-in-first-out” stock

rotation, anoxic protective atmospheres, low storage temperatures, pest-proof packages,

repellents and repellent packages etc. Pest prevention of multiplication inside commodity is

realized via using resistant and GMO varieties, or via “immunization” commodity using low-

toxic protectants such as IGRs, dusts, etc.

Pest-proof storage structures prevent the infestation of bulk grain

Cornel S. Adler, Agnès F. Ndomo-Moualeu ........................................................ 177-184 Abstract: Commercial grain storage warehouses for long-term grain storage in northeastern

Germany were rendered pest-proof by sealing all visible openings. The project idea was to

determine the effect of this measure on the occurrence of infestation in a given grain storage.

Each warehouse had a capacity of some 500 to 1900 t. Sealing doors and walls or gaps between

walls and roof did not cause significant changes in grain temperatures or relative humidity. Rapid

weather changes were rather moderate in better sealed structures, but differences between storage

sites were much greater than between sealed and unsealed warehouses. At one site storing a total

of 3000 t of wheat an infestation of the Indianmeal moth Plodia interpunctella (Lepidoptera,

Pyralidae) was observed in September 2014. Four of five warehouses were found infested with

numerous moths but also with various beetles and even neuropteran species (Chrysoperla spp.).

Indianmeal moths were also found in funnel traps with sex pheromones around the storage

structures. Volatiles above the grain mass and in a down-wind plume outside the unsealed doors

were collected with tubes containing activated charcoal and determined by GC-MS. The

warehouse found un-infested was the one that had been sealed. A pressure test, however, was not

successful which was attributed to remaining openings and cracks in the walls of the building.

This finding seems to prove that storage pests can actively be excluded from stored grain by an

insect-proof seal. Prerequisite for a sealed or hermetic storage is low grain moisture content, in

wheat e.g. max. 13 per cent. Grain must also be stored in boxes remote from outside walls, and

windows should be covered to avoid condensation and moisture migration, respectively.

Effect of nitrogen fertilisation on maize weevil Sitophilus zeamais:

wheat preferences, development time and progeny production Pasquale Trematerra, Marco Colacci ................................................................. 185-191 Abstract: Impact of nitrogen fertilisation on the Sitophilus zeamais Motschulsky wheat-food

preferences, development time, and progeny production were studied. The Italian durum wheat,

Triticum turgidum L., variety “Svevo” was cultivated using three different fertiliser regimes,

0 kg/ha, 60 kg/ha, and 160 kg/ha of nitrogen. The kernel samples obtained by the three

fertilisation regime-plots were compared. After bioassays were performed in cylindrical arenas,

different levels of kernel susceptibility to S. zeamais infestation were observed. Results of trials

indicated that the susceptibility of grains is not closely related to higher protein presence in

kernels from field plots fertilised with 60 and 160 kg/ha of nitrogen compared to the kernels of

unfertilised plots. These observations were consistent with the results observed in F1 and F2

progeny. The overall duration of developmental time of S. zeamais, from egg to adult, was also

not influenced by the different nitrogen fertiliser regimes.

Environmental and host seed effects on the development of Adzuki bean weevil,

Callosobruchus chinensis Soon-Do Bae, Hyun-Ju Kim, Bishwo Prasad Mainali, Young-Nam Yoon, In-Seok Oh, Yeong-Hoon Lee, In-Hee Park, Hang-Won Kang ........................... 193-195 Extended abstract

A method for estimating and predicting losses caused by Sitophilus oryzae (L.)

in paddy and polished rice Baba Gana J. Kabir, Gennady A. Zakladnoy ............................................................. 197 Abstract only

“Speedbox” as an effective mini gas generator for phosphine fumigation

Moshe Kostyukovsky, Elazar Quinn, Chen Sela, Anatoly Trostanetsky, Talat Hazan ........................................................................................................ 199-206 Abstract: Phosphine based fumigants are mainly used today for stored product insect pest

control. However, some limitations, such as low temperatures and relatively long exposure time,

limit their uses. In order to overcome these difficulties, a special device, called "speedbox" was

developed by Detia Degesch GmbH, Germany. The Speedbox is consigned exclusively for the

use of Degesch Plates®. The Speedbox is a small waterproof aluminium box containing a heater

and a ventilator that injects and recirculates the phosphine gas in and out of the fumigated space.

This technology was compaered with traditional phosphine formulations, such as aluminium

phosphide tablets and magnesium phosphide tablets and plates. At the rate of 4 g/m3, using the

Speedbox, phosphine concentrations of 200 ppm and 830 ppm were reached 2 h and 8 h,

respectively, following the fumigation, compared with 35-102 ppm and 200-520 ppm with the

other technologies. The accumulative phosphine concentrations using Speedbox were much

higher during 72 h of the fumigation compared with the non-speedbox technologies. As a

consequence of using the Speedbox a high phosphine concentration was reached practically at the

beginning of the fumigation, 100% mortality of all tested insects at all developmental stages were

recorded after only 48 h treatment at the rate of 6 g/m3 or 72 h at 4 g/m

3. The Speedbox enables

us to control all developmental stages of the major stored product insects by shortening the time

of phosphine treatment from 7-10 days to 2-3 days. The Speedbox as a mini gas generator

demonstrates a high efficacy and other advantages compared with the common phosphine

fumigation technologies for stored product insect pest control.

Managing strongly phosphine resistant rusty grain beetle in Australia Hervoika Pavic, Manoj K. Nayak, Gregory J. Daglish, Patrick J. Collins, Andrew W. Ridley, Rajeswaran Jagadeesan, Ramandeep Kaur ............................... 207 Abstract only

Interpreting customer complaints: Heat treatment inhibits melanin formation

in larvae of the Indian meal moth, Plodia interpunctella (Hübner) Ewa Sady, Bogumił Wiśniewski, Stanisław Ignatowicz ....................................... 209-214 Abstract: The manufacturers of food products try to interpret some customer complaints on final

products that are contaminated by insect pests. Question thus arises – if this insect larva found in

food has passed the processing, and was it treated by heat? To answer this question one should

observe the post mortal processes that occur in insect body. Melanisation of insect body after

death results from loss of control over chemical reactions leading to melanin formation. Insects

employ phenoloxidase and dopachrome (decarboxylating) isomerase for melanin biosynthesis. It

is suggested that heat treatment inhibits activity of these enzymes responsible for melanin

formation, and thus reduces or eliminates melanisation of insect body after death. Therefore,

fully-grown larvae of the Indian meal moth, Plodia interpunctella (Hübner) (Lepidoptera,

Pyralidae), were killed by heat (60 °C and 80 °C for 30 sec.), and observed for whole body

melanisation. Control larvae rapidly melanised after freezing and thawing, whereas heat-killed

larvae failed to show typical melanisation. Thus, lack of melanization could be an indicator of

heat treatment of those insects that were found in the final food product. Sometimes people want

to know where and when the contamination of final product occurred: at consumer house, in a

shop, in distribution centre, or in the factory that produces this product. From time to time, an

entomologist is asked "Can you determine if this insect was cooked?”. If it was heat treated, then

contamination occurred at the factory during a processing of raw materials.

Development of Indian meal moth Plodia interpunctella (Hübner)

inside finished products with low moisture content Renata Zorko ..................................................................................................... 215-221 Abstract: The possibility of Indian meal moth Plodia interpunctella (Hübner) development

inside the finished products with low moisture content was investigated. The Indian meal moth

population used in the study was obtained from various moth-infested commodities and

maintained in laboratory conditions. The nutrient media was the salted powdered mixture which

contained dried minced vegetables. The concentration of salt in the mixture was 50% minimum,

and concentration of dried vegetables was 10% minimum. Maximum concentration of moisture

in the food did not exceed 2%. The samples were divided in four categories in relation to

different packaging materials and different quantity of packaged food. The samples were infested

with 10 eggs per sample and incubated at two temperatures, 30 °C and 22 °C. Three samples of

each category and from each temperature were opened monthly, during the test period of 10

months. The possibility of growth and development of Indian meal moth in the finished product

of salted powdered mixture with dried minced vegetables, in both of tested temperatures, will be

shown. The percentage of egg hatching varied from 72.91% to 94.61%, but the percentage of

development from egg to adult were considerably lower and varied from 0% to 53.97%,

depending on the quantity of packaged food and the incubation temperature. The possibility of

growth and development of Indian meal moth was the highest in the samples with small quantity

of nutrient media, and declined with the increase of the quantity of packaged food. Temperature

of 30 °C resulted in higher percentage of found larvae and adults in food samples, compared to

samples incubated in the room temperature.

Efficacy of Ekomille® in the management of house mouse, Mus musculus Linnaeus,

in wheat facilities in Southern Italy Giuseppe Spina ................................................................................................. 223-230 Abstract: Mus musculus Linnaeus cause significant direct and indirect damages to wheat. The

damages begin already in the field to continue along the food chain. Especially in non-

conditioned stored facilities, rodent population density could reach very high levels. In the

present study are reported field trials carried out to evaluate a non-toxic system, Ekomille® traps,

in rodent management programs. Ekomille® trap is an electromechanical and ecological system

for capture of mice and rats. The trap is baited with toxic-free natural foods and it is able to

capture over than 80 rodents. We selected a wheat facilities situated in Altamura (Southern Italy),

20 silos, 30.000 tons stored, that presents a very high population of M. musculus. Before

Ekomille® disposal, deratization was made with rodenticides in many types of formulations.

However, has been recognized a large number of sightings of mice, especially under the wheat

elevators and have been recorded extensive damages to the electrical systems. From the

beginning of June 2011, 20 Ekomille® baited with natural foods (sunflower seeds, pine nuts and

pork fat) were placed in the wheat facilities, 4 nearby elevator base, 4 nearby electrical

substation, 4 in the second line of defence around silos platform and 8 in the first line of defence

along the external perimeter. Traps have been checked weekly during the first month and

thereafter every two weeks. From the beginning of June 2011 to the end of May 2012, 354 house

mice were captured. After trap use, a drastic reduction of presence of mice and damage caused by

rodent activity was observed. These positive results indicate that use of Ekomille® trap for

rodents control is a valid component of an IPM-based control strategy. In IPM programs, the

employ of these traps can lead to drastic reductions of chemical treatments, resulting in economic

benefits and improvements for food safety and quality.

Improving of acceptability, palatability and efficacy of bromadiolone baits

in controlling Norway rat and house mouse in storage facilities Marina Vukša, Goran Jokić, Suzana Đedović, Bojan Stojnić, Tanja Šćepović ........... 231 Abstract only

Session 3. Prevention of microflora infection and development of mycotoxins Integrated approach of the prevention of mould spoilage risks

and mycotoxin contamination of stored grain – A European perspective Keynote Speaker: Francis Fleurat-Lessard ........................................................ 235-260 Abstract: Stored grain moulds may grow in cereal grain with critical moisture content. The

invasion of seeds by seedborne fungi can induce grain heating, off-odours and reduction of

germination capacity. Some species produce harmful mycotoxins. Mycotoxin contaminations of

cereal grain are a worldwide issue for public health, agro-food industry concerns, and economics.

The most toxigenic storage fungi include members of the genera Aspergillus and Penicillium.

The prevention of mould growth in stored grain bulks is the single way to manage this important

grain quality issue.

The integrated management of storage moulds spoilage risks is based on five pillars:

i/ Prevention of mould development in keeping grain condition below the moisture threshold

enabling harmful fungus species germination and growth; ii/ Accurate monitoring of activity of

water (in grain) and temperature changes during storage, associated to the monitoring of telling

indicators of mould respiration activity; iii/ reduction of grain bulk moistening trends by physical

intervention means or by preservative additives; iv/ use of bio-competitive strains of fungi or

bacteria to prevent the development of Aspergillus spp. and Penicillium spp. toxigenic strains to

install in grain bulks; v/ use of physical treatments (ozone, grain pearling or abrasion) to limit

mycotoxin contamination risks in processed cereal food. Future research needs on this topic are

evocated.

Current status on stored product protection in São Tomé & Principe

Idalina Paquete, Albino Bento, Maria Otilia Carvalho ......................................... 261-267 Abstract: São Tomé & Principe is located in the Gulf of the western equatorial coast of Central

Africa. About half of the country’s total 96,000 ha are used in agriculture. The economy of São

Tomé & Príncipe is dominated by cocoa export, which represents 95% in value of the country’s

revenue. Food-crop importing is essential for population subsistence. Rice consumption requires

6230 tons, with 4132 tons arriving as donation. Customs supervise the state of goods before

harbouring and during storage, mainly in warehouse structures. The methodology adopted is

visual inspection and absolute samples. The present work aimed at evaluating the key-pests of

rice and bean from arrival, during storage until reaching the markets. From January 2004 to

December 2012, 151 rice and 160 bean samples were collected and analysed at the CIAT

laboratory. Samples were obtained from ships and public and private storage facilities. From all

samples, collected only eight rice and 17 bean samples were infested. The year and origin were

reported to identify source of infestation. On rice, the main insects’ species were Sitophilus spp.

and Tribolium castaneum (Herbst). Sitophilus sp. was more abundant and T. castaneum occurred

in all infested samples. Acanthoscelides obtectus (Say) and Bruchus pisorum (L.) and were

identified in bean samples, with 70% and 30% of occurrence respectively. On rice and bean the

major fungi incidence was related with Aspergillus genera. In S. Tome and Principe, due to

equatorial weather conditions, the major risk is related with fungi contamination, namely with

Aspergillus spp., as mycotoxin producer. Insects can play an important role as vectors for fungi

dissemination.

The use of ozone gas for the control of insects and micro-organisms in stored products

Ali A. Işikber, Christos G. Athanassiou ...................................................................... 269 Abstract only

Analysis of mycobiota and oviposition of Callosobruchus maculatus

in beans variety Vigna angularis and Vigna unguiculata Fabricio C. Reis, Thais L. Lima, Marcos Roberto Potenza, Simone Aquino ........ 271-274 Abstract: Beans are an important source of protein in the diet of several societies. In inadequate

storage conditions may occur insect infestations and fungal contamination. This study analysed

different samples of Vigna angularis and Vigna unguiculata, purchased in retail. There were

determined oviposition and viability of Callosobruchus maculatus eggs in adzuki bean

(V. angularis, n = 10), cowpea (V. unguiculata, n = 16) and black-eyed (V. unguiculata, n = 13).

Twenty adult insects with 1-5 days of emergency were confined in 50 grains and assessed the

number of eggs in 8 days and emerged insects to 40 days, with 5 replicates per variety.

C. maculatus oviposited on average 19.4 eggs in adzuki beans, but without viability. In black-

eyed and cowpea egg viability was 31.40 and 13.71%, respectively. Regarding mycobiota, the

direct plating method of grains on Sabouraud agar showed the predominance of Eurotium

chevalieri fungus isolated in 100% of the samples adzuki. The cowpea presented the frequency of

81.2% of yeast and 12.5% of Aspergillus niger. The black-eyed presented the frequency of 100%

of yeast and 61.5% of Aspergillus flavus. It was concluded that the storage conditions should be

monitored in the retail market in the prevention of insect attack and fungal both varieties.

Fungal contamination and insect pests’ assessment in Angola stored maize

Laurinda Paim, Graça Barros, Ana Magro, Margarida S. M. Bastos, Arlindo Lima, António Mexia ...................................................................................... 275 Abstract only

Control of post-harvest insect infestations and microbial contaminations

of Corinth currants and sultanas with infrared radiation Christos G. Athanassiou, Antonia Chiou, Christos I. Rumbos, Andreas Karagiannis, Eirini Nikolidaki, Eirini Panagopoulou, Antonis Kouvelas, Vaios Karathanos ......................................................................... 276 Abstract only

Identification and characterization of Penicillium expansum associated with blue mold of apple during storage in ULO in Croatia Zdravka Sever, Tomislav Kos, Dario Ivić, Tihomir Miličević ................................ 277-283 Abstract: Blue mold, caused by Penicillium expansum is one of the most important postharvest

diseases of apple fruit worldwide. Disease can cause significant economic losses during storage,

as well as losses of processed food commodities due to the production of mycotoxin patulin.

Incidence of pathogens associated with postharvest fruit rots in Croatia was surveyed during two

storage seasons (2009/2010 and 2010/2011) in Ultra Low Oxygen (ULO) conditions and

conventional atmosphere (NA). Diseased fruit (cultivars Idared, Jonagold, Golden Delicious and

Cripps Pink) with blue mold symptoms were collected and 36 Penicillium isolates were

recovered. Identification was based on macroscopic and microscopic features on different media.

Molecular identification was based on PCR with primer pair PEF/PER for specific detection of

P. expansum. Based on morphological and molecular methods P. expansum was detected as

causal agent of blue mold of apples in Croatia. P. expansum was the predominant pathogen of

decayed fruit, accounting for averages of 39.0 and 27.9% on Idared in both seasons and 37.5% on

Jonagold in 2009/2010. On the other hand, during storage in NA yield losses were higher,

P. expansum was detected as minor pathogen, accounting for 2.3 and 3.7%, while brown rot

(Monilinia fructigena) was prevalent disease. It can be concluded that storage conditions impact

pathogens and losses they cause. Since in Croatia, only products containing fungicides boscalid

and pyraclostrobin are registered for control of postharvest diseases of apple, new methods such

as heat treatments, biological control etc. should be implemented in apple production to prevent

fungal diseases and postharvest losses.

Fungal community composition on different potato cultivars during storage

Zane Vigule, Lelde Grantiņa-Leviņa, Ilze Skrabule, Olga Treikale ...................... 285-292 Abstract: The aim of the present study was to investigate the fungal community composition on

different cultivars of potato during storage. The study was performed in 2013-2015 by the

Latvian Plant Protection Research Centre, evaluating disease presence on seed material of potato

tubers after 3 months of storage in storehouses at State Priekuli Plant Breeding Institute and at

several farms in different areas in Latvia. At all places conventional potato cultivation was used,

with differences concerning previous crop, soil conditions and fungicide application. All isolated

fungi were identified based on morphological, cultural, and molecular characteristics using

conventional PCR and sequencing a part of the translation elongation factor 1-α-gene for

Fusarium species and sequencing a part of the ITS1-5.8S-ITS2 region for other fungi. Analysis

of single-conidium isolates of fungi from different cultivars of potato revealed the presence of

Fusarium species causing dry rot of tubers during storage period: F. avenaceum (Fries) Saccardo,

F. crookwellense (syn. F. cerealis) Burgess, Nelson & Toussoun, F. culmorum (W. G. Smith)

Saccardo, F. equiseti (Corda) Saccardo, F. flocciferum Corda, F. oxysporum Schlechtendahl

emend. Snyder & Hansen, F. redolens Wollenweber, F. solani (Martius) Appel & Wollenweber

emend. Snyder & Hansen, Fusarium torulosum (syn. F. sambucinum, F. sambucinum var.

coeruleum, F. venenatum) (Berkeley & Curtis) Nirenberg, F. tricinctum (Corda) Saccardo.

Pathogenic fungi Rhizoctonia solani Kühn, Alternaria solani (Ell. & Mart.) L. R. Jones,

Alternaria alternata (Fr.) Keissler, A. tenuissima (Kunze) Wiltshire and Phoma exigua var.

foveata (Foister) Boerema were found on different potato cultivars. Isolation of Colletotrichum

coccodes (Wallr.) S. J. Hughes (syn. C. atramentarium (Brek. & Broome) Taubenhaus), which

causes black dot of potato during vegetation period, confirmed a presence of seed-tuber-borne

infection in potato seed material. Galactomyces candidum (syn. Geotrichum candidum) de Hoog

& M. T. Sm., which causes rubbery rot of potato was isolated from the samples of affected tubers

from three cultivars. Appearance of soil saprotrophic fungi Trichocladium asperum Harz.,

necrotrophic Plectosphaerella cucumerina (L.) Laterr., Humicola fuscoatra Traaen., Torula

herbarum (Pers.) Link, Volutella ciliata (Alb. & Schwein) Fr. and Microdochium bolleyi

(R. Spraque) de Hoog & Herm.-Nijh. were associated with different growing conditions of potato

cultivars.

Session 4. Natural products Control of stored products pests by natural products

Keynote speaker: Vlatka Rozman ...................................................................... 295-299 Abstract: The use of natural products has been emerging as one of the prime means to protect

crops and their products and the environment from pesticide pollution, which is a global problem.

Natural products possess a spectrum of properties including insecticidal activity, repellence to

pests, antifeedancy, insect growth regulation, toxicity to mites, and other pests of the agricultural

importance. Also they possess antifungal, antiviral and antibacterial properties against pathogens.

More significant use of natural products in practice will be possible if they are compatible with

the following principles: safe, low toxicity, easy for application, minimal (or nil) problems with

insecticide residues on foods, high efficacy at very low concentrations against stored grain

insects, adult pests and their progeny, wide spectrum of efficacy against stored grain insect pests

and field pests, low adverse effects on grain handling and quality properties, and acceptable in

terms of price. Pronounced and multiple synergic modes of action substantially reduce or remove

the need for synthetic chemical active ingredients, which reduces application and incidental

exposure to these chemicals. Demonstrably safer natural products have advantage in the market

where there is a growing aversion to conventional chemical products. In Croatia there is ongoing

research on the development of new formulations of natural insecticides based on inert dusts and

botanical insecticides and their combinations.

IPM approaches for stored date protection in Tunisia:

Emphasis on alternative control methods against the date moth Ectomyelois ceratoniae Zeller (Lepidoptera: Pyralidae) Jouda Mediouni Ben Jemâa ............................................................................... 301-308 Abstract: In Tunisia, dates are an important and valuable export commodity, 16% of the total

value of agricultural exports coming from the date industry. Moreover, dates presented 6.6% of

the total value of agricultural production. Stored dates are subjected to high infestation rates by

pyralid moth pests, mainly the carob moth Ectomyelois ceratoniae (Zeller). This devastating pest

induced significant economic losses during storage. It caused loss of weight and downgrading of

the commercial value of dates. It infested 20% of the harvestable date crop annually. Current

available methods for postharvest control are based on fumigation using synthetic fumigants

mainly phosphine. However, harmful effects of these chemicals on human health and their side

effects on environment, lead to the search of safer and effective alternatives. This paper reported

results of several trials conducted using some natural pesticides derived from plants (essential

oils), microorganisms (Spinosad) and inert dust (Diatomaceous earth) for the control of different

developmental stages of Ectomyelois ceratoniae. Results indicated that these alternatives

exhibited ovicidal, larvicidal and adulticidal potentialities against Ectomyelois ceratoniae. The

four Eucalyptus essential oils tested (E. camaldulensis, E. leucoxylon, E. dumosa and

E. transcontinentalis) displayed strong toxicity against adults and all larval stages.

E. transcontinentalis accomplished 100% mortality of Ectomyelois ceratoniae adults after 2 h of

exposure at the concentration 143 μl/l air. Additionally, E. leucoxylon oil achieved 100%

mortality of fifth instars after 48 h of exposure at the dose 132 μl/l air. On the other hand,

spinosad showed an interesting ovicidal activity against Ectomyelois ceratoniae eggs. At the

concentration 1ppm, 85% of egg mortality was obtained. Additionally, the use of diatomaceous

earth exhibited promising results. No closed eggs were obtained after treating eggs with Protect-

It at the dose of 10 g/kg of dates.

The beer brewing waste spent hops (Humulus lupulus L.) as a valuable source

of repellents useful to ward off insect attacks to stored food Francesca Cosci, Stefano Bedini, Guido Flamini, Barbara Conti ........................ 309-315 Abstract: Hop, Humulus lupulus L., is an aromatic plant largely utilized in the brewing industry

to add flavour and bitterness to beer. As a consequence, a large amount of residual material,

named “spent hops”, is produced by the brewing industry. Since only about 15% of the hop

constituents end up in the beer, spent hops should be further valorised as a low-cost source of

bioactive substances.

In this study, hops and spent hops were evaluated as a source of essential oil (EO) with

repellent activity against two of the main stored food insect pests. Hops and spent hops EO yields

were about 2.4% and 0.1%, respectively. The main components of both the EOs were the

terpenes myrcene, α-humulene, and β-caryophyllene. The main chemical difference between

hops and spent hops was the myrcene percentage. Area preference method bioassays showed that

spent hops EO exerted a strong repellency against the lesser grain borer, Rhyzopertha dominica

(F.) (Bostrichidae) and the granary weevil, Sitophilus granarius (L.) (Curculionidae). RD50

values indicated that R. dominica was about 20 time more susceptible to spent hops EO than S.

granarius. Among the EO components, myrcene was the strongest repellent against R. dominica

while limonene was the most effective compound against S. granarius. However, overall, for its

much higher content, myrcene can be considered the compound that mostly contributes to the

repellent activity of spent hops EO. These results show that spent hops could be an excellent,

virtually costless, source of EO to be utilized as eco-friendly repellent in the protection of stored

food from insect pests.

Neem seed oil – iodized salt mixtures as protectant against Dermestes maculatus

De Geer (Coleoptera: Dermestidae) infesting stored African catfish, Clarias gariepinus Burchell Samuel A. Babarinde, Timothy A. Adebayo, Lamidi A. Usman, Oloduowo M. Ameen, Adeyemi O. Akinyemi, Olamide T. Onajole, Olusegun Olusigo ...................................................................................................... 317 Abstract only

Insecticidal activity of plant methanolic extracts for the control of Sitophilus oryzae,

a stored products’ insect Yasmine Dane, Fazia Mouhouche ............................................................................ 318 Abstract only

Insect growth regulatory bioactivities of essential oils of four medicinal plants

towards different strains of stored grain insect pests Mansoor ul Hasan, Shahzad Saleem, Imran Faraz ................................................... 319 Abstract only

Biocontrol of Callosobruchus maculatus using plant metabolites

Faiza Gueribis, Fabiana Avolio, Nadjia Zermane, Antonio Evidente, Maurizio Vurro ........................................................................................................... 320 Abstract only

Essential oils composition of four Myrtaceae of Tunisia origin

and their biological activity against the cowpea weevil, Callosobruchus maculatus (F., 1775) (Coleoptera: Bruchidae) and Vigna unguiculata seeds Mariam Hedjal Chebheb, Abdellah Kellouche, Larbi Khouja Mohamed .............. 321-328

Abstract: Leguminous plants play a prime role in African populations food supply due to their

high content in proteins and good preservation of the harvests is vital. Callosobruchus maculatus

(Coleoptera: Bruchidae) is one of the most serious pests of leguminous seeds such as cowpea

seed. In several African countries, during storage, many generations thrive and cause losses

varying between 90 and 100%. Our work focuses on the identification of essential oils active

elements of aromatic plants from Tunisia, and their biological activity against cowpea weevil.

The results of gas chromatography coupled with mass spectrometry showed that monoterpene

components predominate over sesquiterpene compounds. Their retention time is shorter

compared to sesquiterpenes. The major component is the eucalyptol. The results of the biological

activity of four essential oils on C. maculatus showed that the application at the dose of

25 µl /50 g of E. lehmani, E. astringens, E. maidenii and E. cinerea gave 100% mortality within

24 h, with no survival embryonic and post- embryonic. However, these treatments did not affect

the seeds germination of V. unguiculata.

Valorisation of the repellent potential of two Tunisian Artemisia essential oils

for the management of the sawtoothed grain beetle Oryzaephilus surinamensis (Coleoptera: Silvanidae) Olfa Bachrouch, Soumaya Haouel, Nadhem Ferjani, Jouda Mediouni Ben Jemâa ............................................................................... 329-332 Abstract: The aim of the current study was to evaluate the repellent potential of two Tunisian

Artemisia essential oils namely desert wormwood Artemisia herba-alba and wormwood

Artemisia absinthium against adults of the sawtoothed grain beetle (Oryzaephilus surinamensis).

Repellency was assessed using filter paper test method. Four doses were tested: 2.5, 5, 7.5 and

10 µl. These doses were calculated to give equivalent repellent concentrations of 0.09, 0.19, 0.29

and 0.39 µl/cm2. Repellency was recorded after 1, 3, 5 and 24 h of exposure.

Results revealed that both essential oils showed repellent potential against adults of this

pest. However, A. absinthium essential oil repellent potential was greater than A. herba-alba

essential oil. In fact, at the lowest concentration 0.09µl/cm2, the percentages of repellency were

75% and 60% respectively with A. absinthium and A. herba-alba after 5 h of exposure.

Significant differences were observed between the two oils for all concentrations and exposure

durations.

Based on these results, wormwood and desert wormwood essential oils have potential for

use as repellents against stored pests in an Integrated Pest Management program in stored

commodities.

Repellency test of essential oils and alcoholic extracts

against Tineola bisselliella (Hummel) Claudia Sotgia, Elisabetta Schiavon, Alessia Berzolla, Elisabetta Chiappini .............. 333 Abstract only

Evaluation of natural compounds for insect repellent packaging:

laboratory and semi-field experiments with pests of cereal products Sara Bortolini, Lara Maistrello ............................................................................ 335-340 Abstract: This work aimed at identifying natural substances with a repellent effect on insect

pests of cereal products, to be used in repellent packaging in semi-field experiments simulating

insect attacks in storage rooms. Initially, the repellence of different concentrations of selected

natural compounds was evaluated on groups of 10 adults of Sitophilus oryzae, Rhyzopertha

dominica, Stegobium paniceum, and 5 larvae of Plodia interpunctella, using Petri dishes with

treated/untreated cardboard disks halves. Then, the performance of repellent cardboard boxes

subjected to repeated mixed insects attacks was evaluated in storage room conditions. The boxes,

obtained by applying paints with microcapsules of essential oils of garlic and/or rosemary (the

most repellent after the first test), were handmade simulating commercial pasta packages and

were filled with 500 g pasta. The boxes were placed in couples (with different combinations that

included untreated control) inside fenestrated plastic containers with groups of 20 insects

(5 adults of S. oryzae, R. dominica, Lasioderma serricorne; 5 larvae of Ephestia spp.). Every

month, insect mortality inside the containers was checked and new groups of insects were

introduced. After 3 months all the cardboard packages were opened and the number of insects

inside and outside the boxes was recorded. The best results were obtained in presence of garlic

essential oil. The criticalities emerged in the view of practical applications are discussed.

Effects of bergamot, lavender and thyme essential oils on the attractiveness of cereal-based baits to house mice Goran Jokić, Marina Vukša, Suzana Đedović, Rada Đurović-Pejčev, Bojan Stojnić, Tanja Šćepović, Saša Vasilev ............................................................. 341 Abstract only

Session 5. Biological control of stored product pests Recent advances in the commercial application of beneficials

against stored-product and cultural heritage pests Keynote Speaker: Matthias Schöller .................................................................. 345-348 Abstract: The fields of commercial application of beneficials against stored-product pests, i.e.

the combination of a certain product, pest and beneficial are analysed. Seventy fields of

application against stored-product and cultural property pests were identified. Applications

include empty room, bulk storage, packaged products, and materials like textiles and artefacts.

The Indian meal moth received moth applications, followed by clothing moth and Sitophilus spp.

weevils. Concerning the natural enemies, the largest number of applications was with

T. evanescens, followed by Lariophagus distinguendus/Anisopteromalus calandrae and

Habrobracon hebetor. Almost the same number of fields of applications was in organic and

conventional stored products. Reasons for these focuses are discussed.

Parasitism by Venturia canescens and Habrobracon hebetor on mono-

and heterospecific populations of pyralid moths located in laboratory and experimental store houses Cristina Castañé, Jordi Riudavets, Eric Lucas .................................................... 349-353 Abstract: Plodia interpunctella and Ephestia kuehniella are important pests of stored products

that are both parasitized by Venturia canescens and Habrobracon hebetor. These two

cosmopolitan larval parasitoids are found in large numbers in food processing facilities in north-

eastern Spain. In laboratory, we evaluated their performance when mono and heterospecific

populations of 3rd

-4th larval stages of the two moths were offered. We also examined the

dispersion capability of females when larvae of the two moths were offered also alone or in

combination. For this purpose hosts were offered in the eight corners of experimental rooms of

aprox. ≈ 30 m3

during three days; the experimental rooms had a window for considering the

influence of natural light illumination on their dispersion pattern. Both parasitoids reduced the

emergence of both moth species by half in the laboratory, in comparison with the controls, either

when offered separately or in combination. Reproduction of H. hebetor was also similar in

presence of both hosts, while V. canescens had a significantly higher reproduction on

E. kuehniella than on P. interpunctella larvae. In experimental rooms both parasitoids were able

to similarly parasitize larvae located in any of the eight corners, independently of the window

situation. However, both parasitoids were more efficient parasitizing E. kuehniella than

P. interpunctella larvae.

Can biological control affect the number of fragments in processed food?

Sara Savoldelli, Daria Patrizia Locatelli, Luciano Süss, Lidia Limonta ................ 355-359 Abstract: Biological control, traditionally considered for field pests, has been recently

considered to control stored products pests, and its efficacy has been demonstrated. The presence

in processed food of fragments derived from biological control agents could be considered a

problem. In this research the results of light filth analysis on semolina and pasta, processed in a

plant where Xylocoris flavipes Reuter (Hemiptera: Anthocoridae) and Anisopteromalus

calandrae (Howard) (Hymenoptera: Pteromalidae) were released to control stored product

beetles, are reported. The aim was to verify if biological control agents can affect the number of

fragments in processed food. Light filth analyses were carried out on semolina and pasta samples

collected before and during the release of predators and parasitoids in semolina processing plant.

The number of fragments detected in the samples collected during the release of the natural

enemies was similar to the ones present in the samples collected before biological control.

Moreover only fragments of stored products pests were detected, fragments directly related to

X. flavipes and A. calandrae lacked. Efficacy of nano-extracted Destruxin from Metarhizium anisopliae

against red flour beetle, Tribolium castaneum, and confused flour beetle, Tribolium confusum (Coleoptera: Tenebrionidae), under laboratory and store conditions Magda M. A. Sabbour ........................................................................................ 361-367 Abstract: The destruxin is a cyclic hexadepsipeptides produced by entomopathogenic and

phytopathogenic fungi five amino acids and one hydroxyl acid. The effect of destruxin and nano-

destruxin on the red flour beetle, Tribolium castaneum, and confused flour beetle, Tribolium

confusum (Coleoptera: Tenebrionidae), under laboratory and store conditions were studied. The

results showed that the mean number of the eggs laid/female of T. castaneum significantly

decreased to 166.6 ± 9.4 and 38.6 ± 9.1eggs/female when treated with destruxin and nano-

destruxin, respectively, as compared to 298.1 ± 9.4 eggs/female in the control. When T. confusum

were treated with destruxin and nano-destruxin the number of eggs/female significantly

decreased to 101.6 ± 4.4 eggs/female and 50.6 ± 9.1 eggs/female when treated with nano-

destruxin as compared to 297.9 ± 5.3 eggs/female in the control. When T. castaneum was treated

with destruxin the number of eggs laid/female were significantly decreased to 31.8 ± 6.9

eggs/female after 120 days. The number of eggs laid/female significantly decreased to 11.8 ± 3.5,

21.7 ± 1.8, 30.1 ± 4.5 and 41.1 ± 1.5 eggs/female after 20, 45, 90 and 120 days when treated with

nano-destruxin. The percentage of adult emergence were decreased to 1, 5, 11 and 19%.

Efficacy of nano-extracted destruxin from Metarhizium anisopliae

against Sitophilus oryzae (L.) (Coleoptera: Curculionidae) under laboratory and store conditions Magda M. A. Sabbour ........................................................................................ 369-375 Abstract: The rice weevil, Sitophilus oryzae (L.) (Coleoptera: Curculionidae), is one of the most

serious stored grain pests worldwide. Destruxin is a cyclic hexadepsipeptides produced by

entomopathogenic and phytopathogenic fungi five amino acids and one hydroxyl acid. The effect

of nano-extracted destruxin from Metarhizium anisopliae was evaluated against the rice weevil.

Results showed that the mean number of eggs laid/female of S. oryzae significantly decreased to

90.6 ± 2.1 and 41.6 ± 3.1eggs/female when treated with destruxin and nano-destruxin,

respectively as compared to 299.6 ± 8.4 eggs/female in the control. When S. oryzae were treated

with destruxin and nano-destruxin the percentage of infestations were significantly decreased to

18 % and 4%, respectively, as compared to 99% in the control in the store.

Role of root border cells and secretions in plant defence

Azeddine Driouich, Koroney Abdul, Maxime Gotte, Youssef Manasfi, Marie-Laure Follet Gueye, Maite Vicré Gibouin ......................................................... 377 Abstract only

Beneficial insects against moth and weevils in long-term grain storages in Germany,

Part I: Comparison of different traps to detect S. granarius Steffi Niedermayer, Solène Juillet, Bernd Wührer, Matthias Schöller,

Sabine Prozell, Johannes Steidle ....................................................................... 379-384 Abstract: Long-term storages have special requirements to stored product protection. Therefore

a three year project was established to improve and adjust existing biological control strategies to

those conditions. To secure an early detection of pests especially of the Granary Weevil

Sitophilus granarius, three different trap types were tested: plastic cups used as simple pitfall

traps, pitfall cone traps with lids, and probe traps. The experiments were conducted in barrels,

BigBags and silo bins. In all setups the probe trap performed best, being more sensitive to low

infestation rates and infestations with O. surinamensis. Furthermore probe traps are not affected

when grain is moved. Therefore, based on our results, probe traps should be the trap of choice.

A qPCR method to detect and quantify Rhyzopertha dominica in brown rice

Mireia Solà, Jonathan G. Lundgren, Nuria Agustí, Jordi Riudavets ........................... 385 Abstract only

Molecular approaches based on PCR to identify and quantify

arthropod internal feeders in grain Mireia Solà, Jordi Riudavets, Nuria Agustí ......................................................... 387-393 Abstract: Insect occur at each step along the chain of food production, leading to serious

quantitative and qualitative losses and representing a major threat to food industries. Albeit

several methodologies to detect stored pests have been developed, they present several

drawbacks. Along the last decades, there has been a global trend in biological studies to use

molecular approaches for diagnosis analysis. Although food industries have been using

Polymerase Chain Reaction (PCR) diagnostics for food control it has not been used yet to detect

insect infestations. We present different molecular approaches based on PCR methodologies to

detect, identify and quantify the presence of insects in raw materials and processed food.

Conventional PCR has been used for the detection of all Rhyzopertha dominica (Fabricius)

(Coleoptera: Bostrichidae) developmental stages in brown rice, wheat and maize even hidden

inside the grain kernels. Multiplex PCR is a variant of conventional PCR permitting simultaneous

detection of different target species. We have developed this method for the detection and

discrimination of the three morphologically similar species of the genus Sitophilus [S. granarius

(L.), S. oryzae (L.), S. zeamais (Motschulsky)] potentially present in stored grain facilities. On

the other hand, decreasing amounts of R. dominica DNA in brown rice have been analysed by

real-time PCR to be used for the quantification of this insect species in food. The introduction of

these molecular approaches in stored grain and food industries will be very useful for the

diagnosis of insect pest species, particularly to reveal infestations of insect immature stages

hidden inside the grain.

The use of the parasitoids Anisopteromalus calandrae and

Lariophagus distinguendus for the control of the cowpea seed beetle Callosobruchus maculatus Ouarda Benkhellat, Jean Paul Monge, Aissa Moali ............................................ 395-399 Abstract: The bruchid Callosobruchus maculatus is the major constrain in cowpea production

and storage in Algeria. Damage due to this insect affects especially the quality, the quantity and

agronomic value of the product. In storage C. maculatus is currently parasitized by two

parasitoids Anisopteromalus calandrae and Lariophagus distinguendus. The aims of this study

were to evaluate the impact of the two parasitoids used alone or simultaneously and at different

density on the reduction of emergence of C. maculatus bruchids adult’s. The effect of intra and

interspecific competition between parasitoids was also studied. The results show that

A. calandrae was much more efficient at reducing of emergence of C. maculatus (32.85%) than

was L. distinguendus (42.92%). Competition reduced emergence of both A. calandrae and

L. distinguendus; however A. calandrae was clearly the dominant species when bruchids were

exposed to equal number of both species of parasitoids.

Mortality effects of selected native Beauveria isolates on three coleopteran pests

of stored wheat under controlled conditions Mehmet Kubilay Er, Ali A. Işikber, Hasan Tunaz, Arife Öz, Fadime Aydin, Ümmühani Tuğba Özbalikçioğlu ................................................................................ 401 Abstract only

A banker box to improve the impact of Habrobracon hebetor on stored product insects Eric Lucas, Jordi Riudavets, Cristina Castañé .................................................... 403-407 Abstract: The treatment of store room walls, ceilings, floors and other structures is necessary as

part of the good hygienic procedures recommended for reducing pests problems. Structural pests

such as some moth species hide in the floor, corners, cracks and crevices inside machinery and

are always difficult to control. With the continuous reduction of the availability of the

insecticides, it is relevant to evaluate potential alternative methods of control. Habrobracon

hebetor is a gregarious ectoparasitoid of pyralid moths that is found in large numbers in several

food processing facilities in north-eastern Spain. H. hebetor can be a good candidate for

biological control of moths that contaminate structures. In this work, we developed a banker box

system for rearing H. hebetor on Ephestia kuehniella larvae and for releasing them progressively

in the room. Different host-parasitoid ratios were tested to optimize the efficiency of the rearing

box. Also, different apertures in the box were tested in order to allow the exit of adult parasitoids

but avoid the larval moths to escape.

Biological and molecular analysis of an isolated Granulovirus, Tecia solanivora,

for protection of stored potatoes collected in the Andes of Venezuela Alonso Arroyo, Laura Niño, Artiom Carmona, Paulo Beserra Maia, Gerardo Medina, Xavier Léry, Miguel López-Ferber .................................................. 409 Abstract only

Seed-applied bioprotectants for control of seedborne Alternaria arborescens

and growth enhancement of wheat Analía Perelló, Gladys Lampugnani, Cecilia Abramoff, Cecilia Fusé, Gustavo Dal Bello .............................................................................................. 411-415 Abstract: Alternaria arborescens was recently detected as a new member of the Alternaria spp.

complex causing black point in Argentina. Seed treatments with Trichoderma harzianum and

Epicoccum nigrum; Lippia alba and garlic extracts; sodium bicarbonate (SB), salicylic acid (SA),

potassium chloride (PC) and sodium phosphate dibasic (SP) were applied to grains of wheat cv

BIOINTA 1004 previously to the infection with the pathogen following ISTA (blotter method)

rules. After 7 days, seed germination, infected seeds, necrotic symptoms of emerged seedlings

and fresh weight were evaluated. Remarkable results were obtained with L. alba, SA and SP that

reduced symptoms from 50 to 78% compared with the control. Necrosis of radicles was

significantly reduced by the application of all treatments tested. Moreover, fresh weight of

seedlings was significantly increased with the application of the two antagonists, garlic and the

three tested salts in comparison with control. It is concluded that compounds here tested have

potential as biofertilizers and ecofriendly alternatives to control seed-borne fungi of wheat.

Postharvest control of Botrytis gray mold in tomato by antagonists

and biorational compounds Gustavo Dal Bello, Gladys Lampugnani, Cecilia Abramoff, Cecilia Fusé, Analía Perelló ..................................................................................................... 417-425 Abstract: The fungal pathogen Botrytis cinerea causes severe rots on tomato fruit during storage

and shelf life. Continuous use of synthetic fungicides has produced an increasing public concern

regarding contamination of edible crops with toxic residues, and proliferation of resistance in the

pathogen populations. These facts have led to a search for new alternative control measures in a

more ecological way. This study evaluates efficacy of biocontrol agents (Trichoderma harzianum

and Epicoccum nigrum), botanicals (Lippia alba and garlic extracts) and low-risk chemicals such

as salicylic acid (SA), sodium bicarbonate (SB), potassium chloride (PC) and sodium phosphate

dibasic (SP) against gray mold caused by B. cinerea on postharvest tomato fruits. The fruits were

obtained in packinghouse, being washed and disinfected with sodium hypochlorite. Artificially

wounded fruits were treated sequentially with each bio-agent and the pathogen. After 1 week of

incubation in plastic trays, the soft rot lesion diameters were evaluated. Results obtained have

demonstrated that the treatment with T. harzianum, E. nigrum, PC, SP and garlic extract reduced

significantly the gray mold infections from 95 up to 99% compared to the control treated with

B. cinerea alone. These findings indicate that these environmentally friendly agents have the

potential to control postharvest gray mould on tomato fruits in an integrated pest management

strategy.

The influence of native isolates of Metarhizium anisopliae on mortality of

Blattella germanica adults co-injected with eicosanoid biosynthesis inhibitors Hasan Tunaz, Mehmet Kubilay Er, Ali A. Işikber ....................................................... 427 Abstract only

Studies on the multiple host range, developmental compatiblity

and biointensive management of Callosobruchus theobromae (Linnaeus) (Coleoptera: Bruchidae) Desh R. Thakur ......................................................................................................... 428 Abstract only

Session 6. Wood-boring, urban and museum pests Basic research and defence against wood-destroying pests

Keynote speaker: Elisabetta Chiappini ............................................................... 431-435 Abstract: Cultural properties made from organic materials are easily damaged by biotic agents

of different nature (insects, fungi, bacteria). In particular, wood artefacts are often attacked by

insects which belong to taxa very distant from each other: basically that of Isoptera and that of

Coleoptera. Furthermore, in the latter group, families that evolved along very distant lineages, i.e.

powderpost beetles, anobiids and longhorn beetles, adopt various strategies to feed on wood. The

functional morphology, as well as the taxonomy, the behaviour and ways of life, and the modes

of attack turn out into different damages, different defence options and different risks. Therefore,

a thorough and comprehensive knowledge of these biodeteriogens, mainly woodborers and

termites in our case, as regards everything they are concerned with in their life, is essential, for

defence, particularly when considering it as prevention. Many applied researches and, even

worse, many common defence actions are not based on reliable data obtained from basic research

but on hypothesis, common sense and suppositions with the obvious consequence of not being

effective and so invalidating the sustainability of preventive conservation for the protection of

cultural heritage. Monitoring, risk assessment and control, all need a deeper knowledge to be

cogent. Examples are given.

Intraguild competition between two larval parasitoids

of the webbing clothes moth Tineola bisselliella and possible impact on control strategies under laboratory conditions Rudy Plarre ............................................................................................................... 437 Abstract only

Efficacy of ethanedinitrile (EDN) against wood-boring pests

Swaminathan Thalavai Sundaram, Adam Jonas, Yonglin Ren .................................. 438 Abstract only

Temperature and relative humidity effects on Stegobium paniceum (L.)

(Coleoptera: Anobiidae) in controlled atmospheres Elisabetta Chiappini, Alessia Berzolla, Claudia Sotgia ....................................... 439-443 Abstract: Key ecological factors such as temperature and water influence the distribution and

population dynamics of terrestrial arthropods, so the speed of the anoxia treatment in killing

insects is dependent upon temperature, relative humidity (RH) and on oxygen percentage. The

present study reports on the efficacy of temperature and relative humidity on controlled

atmosphere treatment, as an alternative to insecticide management, against the drugstore beetle

Stegobium paniceum (Linnaeus), one of the most widely encountered insect causing serious

damages to stored products and cultural heritage. The aim of the research is also to verify the

feasibility of anoxia at percentages of oxygen significantly higher than those normally used (<

1%), in order to define the application conditions of the method, easier and less expensive.

Larvae and adults were exposed to atmospheres containing low oxygen percentages (3-5%), at

four different relative humidities (15% – 35% – 55% – 75%) and two different temperatures (23-

30 °C) for periods of 7 days. Temperature affects S. paniceum, mainly the adult stage. They seem

to be more susceptible, while larvae appear very tolerant, showing a modest mortality at 23 °C,

with no difference in values. Only at 30 °C and low humidity (dry conditions) they reach 70-80%

of mortality, with significant difference between the worst and the “best” humidity condition (15-

75%). Adults show a higher mortality compared to the larvae, however it seems that there are no

differences between the relative humidity percentages, except at 5% oxygen percentage. The

important role of controlled atmospheres can be improved by studying the sensitive relationships

between these environmental parameters critical to the survival of insects and testing more

possible combinations to achieve the highest efficiency in less time and at lower cost.

Comparative efficacy of neem Azadirachta indica A. Juss, extracts

against powder-post beetle, Lyctus africanus Lesne attacking seasoned wood in Egypt Ahmed Merghem ............................................................................................... 445-454

Abstract: The present study deals with one of the most economic important wood borer; the

powder-post beetle Lyctus africanus Lesne (Coleoptera: Lyctidae) which devastates wooden

production in industry, forestry and agriculture including timber, bamboo and stored products on

both local and global distribution scales. In Egypt, such as in many countries worldwide, control

programs of this dangerous pest are restricted to the usage of chemical pesticides. Thus,

alternative methods with lower risk of toxicity as neem Azadirachta indica A. Juss (Sapindales:

Meliaceae) extracts are encouraged. Through this work, three crude extracts of neem seeds were

used against this lyctid borer on seasoned wood cuttings. These extracts were generated from the

crude by three solvents apart; petroleum ether, ethanol, and distilled water. A second experiment

series for comparison using a commercial neem product "Nemazal" in addition to a reference of

standard recommended insecticide "Cidial" were conducted. Experiments were designed to apply

protective and remedial techniques against L. africanus attacks by recording the mortality and

repellency rates as indicators under natural laboratory conditions. Each extract was tested in three

concentrations; 6, 3, 1.5 gm/100 ml for each separate solvent whereas the commercial products

were used with their recommended dosages (3 ml/l). Experiments revealed that petroleum ether

neem extract in a concentration of 6 gm/100 ml resulted in the highest significant average

mortality and repellency rates with L. africanus reached 81.7 and 76.3% at P > 0.05, respectively

whereas distilled water neem extracts showed the lowest infestation levels with this lyctid borer

at P > 0.05 recording a least average mortality rate of 17.4% and 21.8% repellency rate. These

gained results were confirmed by lethal times and toxicity lines which were estimated by the log-

time probit model. On the other hand, the commercial products of Nemazal and Cidial resulted in

significant higher mortality and repellency levels at P > 0.05 compared with those of the crude

neem extracts. Nemazal and Cidial titres scored 90.3 and 97.1% average mortality rates

meanwhile the average figures of repellency levels were 87.3 and 93.6%, respectively. A new biological control method for the common furniture beetle,

Anobium punctatum Alexander Kassel, Judith Auer ........................................................................... 455-461 Abstract: The common furniture beetle (Anobium punctatum) is a permanent threat in churches

and museums, is wreaking devastating damage by corrosion of wooden inventory and antiques.

The conventional control methods are complex, polluting and high-priced. The aim of our

research was to develop and establish a biological control method for the common furniture

beetle with the braconid wasp species Spathius exarator, the most common antagonist of

A. punctatum. After successful mass rearing, laboratory and praxis tests of the parasitism rate and

the predator-prey relationship were performed by documentation of newly appeared exit holes

from wasps and furniture beetles over time. The laboratory control tests proved 80% parasitism

by the braconid wasps after three and 98% parasitism after ten months. The results of the

practical tests in churches and museums (n = 13) revealed an average predator-prey relationship

of 3 A. punctatum in the first, 0.37 A. punctatum in the second and 0.13 A. punctatum per

S. exarator in the third year of treatment. In contrast untreated objects (n = 25) showed an annual

average relationship of 26.5 A. punctatum per S. exarator. The results demonstrate the biological

control of the common furniture beetle with the braconid wasps as an efficient, sustainable

alternative to conventional, polluting methods. However, the development of pest infestation in

the treated objects remains to be seen in the following years.

Effects of chewing insect attack (Blaptica dubia) on colour photographic prints

Marianna Adamo, Giuseppe Cotellessa, Massimo De Francesco, Donatella Matè, Pasquale Trematerra ................................................................ 463-473 Abstract: In the context of a larger research aimed to highlight possible differences in the

vulnerability of photographic materials related to different manufacturing characteristics (i.e. the

three main binders: albumin, collodion, and gelatin) several trials have been carried out to assess

the damage caused by Blattodea, omnivorous insects widely distributed in museum and in

archives environments. They are particularly harmful to paper materials because of their ability to

corrode and mess, with their excrements, the objects with which they come in contact; moreover

they can provoke public health problems (allergies) and sometimes transmit infectious diseases.

In this paper we present the results of a study carried out to investigate the damage caused by

Blaptica dubia (Blaberidae) to coloured photographic materials samples such as: chromogenic

colour prints (resin coated), photomecanical prints (offset) and inkjet prints, exposed to insects

and monitored for a three weeks period. XRF Spectroscopy was used to perform elemental

analysis of samples. Radiation technologies can be considered an alternative method to ethylene

oxide disinfestation.

Session 7. Quarantine and regulatory issues The European Standard 16636 for pest control services:

a new step toward higher professionalism of the operators in order to protect the consumer’s rights Keynote speaker: Sergio Urizio .......................................................................... 477-482 Abstract: In 2008, the World Health Organisation (WHO) reported that the last sixty years had

brought significant changes in ecology, climate and human behaviour that favoured the

development of urban pests. In the meantime, the food industry and the providers of professional

pest control services were beginning to feel the need to comply with the EU Regulations that

applied and the types of control systems used in the food sector: it was becoming clear that there

was a need to unify and further professionalize the European Pest Management Industry.

Simultaneously, in 2008 CEPA, the European Confederation of Pest Control National

Associations, held in Rome the 1st Convention EUROPEST where “The Roma Protocol” was

issued, establishing the basic rules for a Common Training program for Pest Control Operators.

In 2009, the CEPA Board decided for the application of a CEN Standard for Pest Control

Services and CEN-CENELEC admitted the application as TC/404, sponsored by the Italian

ANID (Associazione Nazionale delle Imprese di Disinfestazione). It was clear to all operators

that it was necessary to develop a single common essential standard throughout Europe. The

objective of the standard should be to show to clients that service providers following this

standard offer a quality recognized as being truly professional within all Europe and would

ensure that its operators were competent to sell and deliver the service they were offering by

meeting specified and validated minimum standards of knowledge, skill and practical

competence. European Standard EN 16636 was approved by CEN on 10 January 2015, after

more than 5 years of work and meetings with all European Countries. The goal of every pest

management activity is the effective and economical reduction or elimination of damage caused

by pests. The successful achievement of this goal depends on a partnership approach between the

service provider and the client. To achieve this goal, the professional service provider conforms

to the principles established by the WHO in respect of Integrated Pest Management (IPM), which

can include a combination of habitat modification, biological control, physical control, chemical

control, environmental impact as well as animal welfare. The key contents of EN 16636 are

essentially two: the process of professional service and the competences of the operators.

Emerging global technological challenges

in the reduction of post-harvest food losses Shlomo Navarro, Hagit Navarro, Simcha Finkelman .......................................... 483-491 Abstract: The adverse effects of pesticide residues in food and the environment resulted in

imposing strict limitations on pesticide registration by regulatory agencies. On the other hand

consumer demand for chemical-free and insect contamination-free products is a general tendency

with which the food industry finds it difficult to conform. This paper reviews the emerging global

technological challenges in the reduction of post-harvest food losses. In many countries, storage

insects like Rhyzopertha dominica, Tribolium castaneum and Sitophilus oryzae have been

reported to develop resistance to contact insecticides and to the conventionally used phosphine

gas. Phosphine fumigation is a common treatment where three important points deserve attention:

a) sufficient gas tightness, the lack of which leads to insect resistance; b) recirculation to obtain

uniform gas concentration; and c) sufficient exposure time for complete control. Other registered

fumigants suffer from the limitation that they may be useful for application using special

equipment or under specific conditions. The most common non-chemical alternative for storage

of cereals is the use of aeration systems during the winter and refrigerated aeration in the

summertime with the objective to achieve temperatures of less than 18 °C to reduce insect

activity. In the summertime, refrigeration provides an excellent solution for quality maintenance

of grain. However, aeration has a limited effectiveness in tropical climate areas. Field trials have

demonstrated the efficacy of thermal disinfestation in flourmills and in the dry fruit industry,

particularly for dates. Other gaseous treatments that have successfully replaced fumigants are the

manipulation of modified atmospheres (MAs) through the use of biogenerated MAs and hermetic

storage, for insect control and for quality preservation of stored cereals, cocoa beans, and high

moisture corn. MA has niche application because of its higher cost than conventional fumigation

using phosphine.

Quick phosphine fumigation technology in gastight structures

for fresh plants treatment Simcha Finkelman, Shlomo Navarro, Hagit Navarro ................................................. 493 Abstract only

Interpreting customer complaints: Defective or poorly designed packaging material

that enables penetration of product by larvae of the Indian meal moth, Plodia interpunctella (Hübner) Ewa Sady, Stanisław Ignatowicz ........................................................................ 495-497 Abstract: Many customer complaints are on food products contaminated by insect pests. The

problem arise from holes in the package, seals that did not seal properly, sealing of the packaging

material with product parts, boxes that do not have an inner liner, and even packaging material

that can be chewed into by insect pests. Also, from openings produced by manufacturers to

remove the excess of air or to facilitate rice cooking. Food aroma/volatiles escaping from

openings, defective sealing or damaged packaging material attracts pests for feeding. Recently

emerged larva (L1) from its egg is an invasive stage of the Indian meal moth, Plodia

interpunctella (Hübner). These tiny L1 do not penetrate unbroken packaging, but efficiently

locate small openings to infest packages. Invasive L1 could gain entry into the packaged products

through minute holes and defective sealing of the packaging material. Manufacturers rethink their

packaging design and try to improve their packaging by correcting of foil sealing process or by

using alternatives.

Session 8. Future trends on all aspects of storage pest control Future trends for stored product pest control

Keynote speaker: Maureen Wakefield ............................................................... 501-509 Abstract: The control of stored product insects and mites relies heavily on the use of chemical

pesticides. Concerns over pesticide residues in food and the environmental impact of some of the

chemicals used prompted research for alternative methods to protect stored crops from

infestation. Chemical, physical and biological methods have been studied. Best practice in the

UK currently uses an integrated approach for invertebrate pest control with physical methods

(cooling and drying) a core component of the strategy. More recently changes to pesticide

legislation in the European Union and concerns over resistance of insects and mites to commonly

used insecticides have added to the need to develop alternative strategies. Significant

developments have been made in some areas, particularly for control of insects using

entomopathogenic fungi. This research has progressed from the initial identification of an isolate

of the entomopathogenic fungus Beauveria bassiana through laboratory testing and field scale

applications, culminating in the EU Annex 1 submission by Exosect Ltd. in 2014. In this

presentation the latest developments in this research will be reviewed and the prospects for other

novel control methods for the future will be discussed.

Screening of reproductive symbionts of Sitophilus granarius, Sitophilus zeamais

and their parasitoid Lariophagus distinguendus Aydin Suzu Tuncbilek, Sevgi Bakir, Ilhan Derin, Hasne Bilbil ............................. 511-517 Abstract: Arthropods are frequently infected with several micro-organisms, including symbiotic

bacteria. They can have a broad spectrum of effects on their hosts, ranging from reproductive

manipulations such as cytoplasmic incompatibility, feminization of genetic males,

parthenogenesis and embryonic male-killing. The diversification and popularization of molecular

tools have led to the increasing use of molecular techniques to identify symbionts. The genus

Sitophilus encompasses species of great economic importance as stored grain pests worldwide.

Among these species, the granary and the rice weevils (Sitophilus granarius and Sitophilus

zeamais, respectively) are particularly important. Sitophilus weevils are hosts to many

endosymbiotic bacteria. Weevils provide symbionts with a stable environment and some

metabolites, while the symbiont provides the weevil with nutrients deficient in their diet. In this

study, we have screened for presence of the intracellular symbiotic bacteria of the S. granarius,

S. zeamais and the parasitoid Lariophagus distinguendus by molecular technique. Here, we

present evidence of Wolbachia between the granary weevils S. granarius and its parasitoid

L. distinguendus. Spiroplasma was found only in L. distinguendus and Arsenophonus in

S. granarius.

Diagnostic method to determine infestation of Adzuki bean weevil,

Callosobruchus chinensis on Adzuki bean seeds Hyun-Ju Kim, Soon-Do Bae, Bishwo Prasad Mainali, Young-Nam Yoon, In-Seok Oh, Yeong-Hoon Lee, In-Hee Park And Hang-Won Kang ..................... 519-520 Abstract only

Molecular characterization of Wolbachia strains associated with grain weevil

Gislaine A. Carvalho, Alberto S. Corrêa, Luiz O. Oliveira, Raul N. C. Guedes, Marcela S. Rodriguero ............................................................... 521 Abstract only

A neglected potential? Volatile organic substances (VOCs) in stored-product protection Tina Gasch, Andrea Krähmer .................................................................................... 522 Abstract only

Bio-efficacy of albumin 1b of legume seeds against granary weevils

Abdelkrim Mebarkia, Frédéric Gressent .................................................................... 523 Abstract only

Qualitative analyses of khapra beetle (Trogoderma granarium Everts)

using real-time PCR Satoshi Furui, Akihiro Miyanoshita, Taro Imamura, Yasutaka Minegishi, Ryota Kokutani .......................................................................................................... 524 Abstract only

Variation in pheromone content of Indian meal moth glands

and emissions with age and time of night Charles S. Burks, Lodewyk S. Kuenen, Mathew Hicks .............................................. 525 Abstract only

Host-mediated larval competition in the Mexican bean weevil

Zabrotes subfasciatus (Chrysomelidae: Bruchinae) Sharrine Omari D. Oliveira, Alice S. Rodrigues, Juliana L. Vieira, Conrado A. Rosi-Denadai, Nelsa Maria P. Guedes, Raul N. C. Guedes ................... 526 Abstract only

Growth performance and chemical composition of larvae of Tenebrio molitor L.

(Col.: Tenebrionidae) Eirini Anastasaki, Kyriakos Aggelakopoulos, Dimitrios C. Kontodimas ............... 527-530 Abstract: Mealworm (Tenebrio molitor L.) is a stored product pest of which larvae are used as

food for birds and reptiles pets as well could be used for the production of fodder for livestock.

The aim of this study was to investigate the growth of larvae and their chemical composition after

feeding on a mixture of bran and potato tubers. Larvae were obtained from a rearing carried on at

constant conditions (25 °C, 65% R. H. and 16 h L/8 h D photoperiod). In these conditions, the

mealworm larvae increased their weight 36 times during a 30-days period. Larvae contained

63.7% of water, high levels of total protein at 11.2%, and 14.5% of total fat. Oleic acid (C18:1),

palmitic acid (C16:0) and linoleic acid (C18:2) were the predominant fatty acids compromising

almost 90% of total lipid content. Larvae of mealworm had a fast growth rate and are good

source of protein and mono and poly unsaturated fatty acids.