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DEVELOPMENT OF NEW ECOLOGICAL PESTICIDES BY INCORPORATION OF SYNERGIC BIO MOLECULES

Final Report Summary - BIMOSYN (DEVELOPMENT OF NEW ECOLOGICAL PESTICIDES BY INCORPORATION OF SYNERGIC BIO MOLECULES)

Executive summary

The European Commission’s Seventh Framework Research Programme (FP7) research project BIMOSYN was successfully finished in February 2013. It focused on studying synergic, active plant extracts to enhance the efficacy of customarily used biocides in biocide products for urban pests and wood protectors (preservatives). The six technical work packages below show the main results that were obtained.

WP1: supply of raw material

A variety of plant extracts were selected and produced for the initial studies using a Soil Medium Extract (SME) approach . Two types of extracts, one from antioxidant-type plants and the other from medicinal-type plants, were investigated. Insecticides and fungicides were also selected to assist with chemical development and formulation.

WP2: study of synergic effects

1. Two synergic candidate extracts were found in screening tests that improved the knockdown effect of sprayed insecticide against houseflies and cockroaches.
2. The optimal synergic ratio of extract to insecticide was analysed
3. Also, two pure compounds were found to cause this synergic knockdown effect
4. In the screening of wood resistance against fungal degradation, two other purified extracts showed synergy with fungicides. These extracts also gave synergy with one insecticide against termites.

WP3: formulation of pesticide products

1. Prototype formulations were developed by two SMEs: one microemulsion concentrate for spraying against urban pests and one diluted microemulsion for the protection of wood.
2. Results were produced about physical and chemical stability of both prototypes and the main problems were identified for further improvements.

WP4: standardised biological and ecotoxicity laboratory tests

1. Several wood-protection prototypes with reduced fungicide content fulfil standard testing against fungal degradation (EN113), testing after aging by leaching (EN84) and evaporation (EN73), and resistance against blue stain fungi (EN152). The wood formulation prototype with insecticide still needs optimisation as they failed against termites (EN117 and EN118) after water leaching (EN84). Anyhow, these formulations passed the termite test after thermal evaporation (EN73), and also passed the test against Hylotropus larvae (EN46).
2. Sprayable insecticide prototypes were tested in knockdown assay against houseflies. These water-based prototypes mainly failed in reproducing the synergic effect found in screenings during WP2. One formulation gave a slight synergic effect and could be used for further development.
3. Ecotoxicity tests on fresh-water organisms were performed with the two selected extracts used for formulation of pesticide products in WP3. The extract used for wood-protective formulations was found to be harmful to fish and toxic against daphnia and algae. Anyhow, the possibility to reduce the content of even more toxic synthetic biocides in wood preservatives was evaluated in WP5. The extract used for sprayable insecticide to improve the knockdown has been evaluated as not toxic to the tested water organisms.

WP5: industrial validation

1. Following semi-industrial evaluation of protective formulations for wood, larger scale impregnation of timber showed a good impregnation of extract and reasonable penetration of fungicide up to 20 mm.
2. Real case treatments against insect pests confirmed that a surface application of one synergic insecticide prototype formulation was effective against cockroaches.
3. With regard to cost analysis, the addition of synergic extract to the wood protector would increase the price, although the reduced biocide content would be beneficial to the environment and to public opinion. In the case of the sprayable insecticide, it was evaluated that the price would actually be increased too much and further development on the extraction would be needed to solve this problem.

WP6: diffusion%%This WP has been defining the protection scheme for knowledge gained from the research, among the SMEs and regarding diffusion to wood-protection and urban-pest sectors.

Project context and objectives

The Spanish Institute Tecnalia, through its Construction Unit, has been participating in and coordinating an FP7 research project for small and medium-sized enterprises (SMEs), called BIMOSYN. The aim has been to develop pest-control products and wood preservatives with lower concentrations of biocides that are therefore more environmentally friendly and less damaging to human health. The two-year project has been carried out jointly by the Swedish University of Agricultural Sciences (SLU) and five other European SMEs.

The scientific development of the project has involved research into the degree of synergy between plant antioxidant substances (natural extracts) and medicinal plant extracts, and conventional insecticides and fungicides. The aim has been to use natural, vegetable-origin extracts with the potential to be employed as biocide adjuvants for the chemical sector. In this way, pest-control products and wood preservatives that will have the same efficacy as conventional ones can be obtained but, at the same time, these will be less prejudicial for the environment and to human health. The new formulations have the potential to combat plagues of flies, cockroaches, termites and wood-rotting fungi.

The main objective of this project has involved studying the synergism between antioxidant bioactive molecules of different types and synthetic biocides included within the European Biocidal Products Directive (BPD) 98/8/CE, for the development of new pesticide products whose performance features may clearly help to distinguish them from other products available on the market. Using this strategy, the aim is for the resulting anti-pest product to be competitive both economically and effectively with regard to those existing on the market, whose biocidal characteristics are unquestionable, albeit problematic due to their high level of toxicity.

As part of the project, a search was carried out for natural antioxidant and medicinal plant extracts that possess no insecticidal or fungicidal properties on their own, whereby they cannot be considered to be biocides as such. This has been the innovative reasoning behind this project: synergy mechanisms may be generated in combination with common pesticides, which can increase the toxic potential of the biocides, thus enabling the concentration of pesticides to be reduced in the final formulation. In this way, a pesticide product will be obtained with far more environmentally friendly characteristics that are also less harmful for operators, at the same time as lowering the cost of the formulation. This means that the pesticide product resulting from this study will be far more competitive for the sector.

Scientific and technological objectives

1. to study the preventive pest-control effect of ecologically developed formulations by comparing them with conventional chemical products;
2. to increase the biocidal effectiveness of pest-control substances by studying the synergy mechanisms between antioxidant molecules or medicinal plant extracts and synthetic biocides included within the directive, with a substantial reduction in the percentage of biocide customarily used.

Environmental objectives

1. to avoid or reduce the use of conventional toxic biocides like pyrethroids and neonicotinoids which are harmful to human health and the environment;
2. to promote the treatment of wood and timber using environmentally-friendly biocide products in construction, civil works and infrastructure sectors as well as the agricultural sector, as opposed to wood materials treated with highly toxic products or other materials that are far more energy-consuming and that result in a negative atmospheric carbon balance, such as steel and concrete;
3. to reduce air, ground and underground water pollution caused by emissions being released into the atmosphere and the leaching of highly toxic biocide compounds.

Socio-economic and industrial objectives

1. to increase the added value of food and agriculture residues used for synergic extracts over other ones which are currently being used that are less profitable and interesting;
2. to develop more economical and environmentally friendly biocide products than current ones by substantially reducing the concentration of biocide and, therefore:
(a) reducing the costs of treatment and offering more competitive pesticide products on the market;
(b) reducing costs related to legal and environmental requirements in treatment and in urban-disinfection processes.
3. to provide new substances, that may enable manufacturers of pesticide products to enter a new market for non-pollutant products that are guaranteed by standardised tests.
4. to help to modernise and adapt the pest industry to coming changes.

Project results

Description of work and main science and technology results

In order to carry out this work, the project was organised according to the following scheme of work packages:
- WP1: Supply of raw material;
- WP2: Study of synergic effect;
- WP3: Formulation of pesticide products;
- WP4: Standardised biological and toxicity laboratory tests;
- WP5: Industrial validation;
- WP6: Diffusion;
- WP7: Project management.

After the first screening of synergic extracts, four positive candidates were found: two for wood protection and two for urban pesticide products. These were evaluated to confirm the synergic effects and to optimise the extract-to-biocide ratios. Work was also performed to identify possible active molecules in the extracts.

Two different types of synergic effect were found.
1. Increased resistance of wood against fungal degradation after treatment with synergic extract from Berkem SA in combination with fungicide. To some extent, synergic effects were also found using insecticide against xylophagous insects.
2. Improved knock-down rate of house flies when combining synergic extract from Rutland Biodynamics Ltd. with several insecticides.

In the second part of the project, one extract for each of these two applications were selected for the formulation of water-based prototype products by the responsible SMEs. The commercial idea was to formulate similar products to those that the companies actually have on the market; that is, concentrated micro-emulsion formulations to be diluted with water before application by professional users. Chemical-stability and biological-efficacy tests were performed on the formulated prototypes. To evaluate the wood-protection prototypes, tests were carried out according to the following European standards: basidomycetic fungi (EN113), blue stain fungi (EN152), termites (EN117 and EN118) and Hylotrupes larvae (EN46). These were carried out after a process of artificial ageing using leaching (EN84) and evaporation (EN73) procedures.

The urban-pesticide formulations were tested by the following described methods: an aerosol spray test against flying insects and a small chamber knock-down test against Musca domestica. The final industrial evaluation included a larger sample treatment of wood and a real case test with biocidal formulation, cost analysis and a toxicity evaluation of prototype formulations.

Main results with the prototype synergic wood protector

1. It was possible to lower the effective concentration of fungicides in treated wood from 150 g/m3 to 100 g/m3.
2. The prototype was effective against wood staining with only 50 g/m3 fungicide.
3. The prototype insecticide formulations still need to be optimised, but in the screenings, without the ageing pre-treatments, a possible 50 % reduction in insecticide was indicated.

Main results with the prototype pesticide formulation

1. One prototype found with improved knockdown on houseflies still needed formulation optimisation.
2. Screening formulations have indicated the potential to reduce the insecticide content by up to 50 %.

The results of the six technical work packages are further described below.

WP1: supply of raw material

A variety of plant extracts were selected and produced by the plant-extraction SMEs for the initial studies, with the potential to work in synergy with biocides. Two types of extracts, one of an antioxidant and the other of a medicinal plant nature, were prepared for the investigation.

Insecticides and fungicides were selected for biocide products for both urban pests (PT18) and for protection of wood materials (PT8), as included or evaluated in the European Biocide Product Directive 98/8/CE (BPD). Other chemicals for development and formulation were also selected.

WP2: study of synergic effect

Initial screening tests were performed to find synergic extract candidates for further investigation in the two areas of urban pests and wood protection.

Urban pests

Two synergic candidate extracts were found in screening tests that improved the knockdown effect of sprayed insecticide against houseflies and cockroaches. The optimal synergic ratios of extract-to-insecticide were analysed, with intent to establish which active molecules were responsible for the synergic effect in the extracts. Two pure compounds were found to cause this synergic knockdown effect.

Wood protection

In the screening of wood resistance against fungal degradation, two other purified extracts showed synergy with fungicides when co-impregnated in wooden samples. These extracts also gave a synergic effect with one insecticide against termites.

WP3: formulation of pesticide products

Prototype formulations have been developed by two SMEs: one microemulsion concentrate for spraying against urban pests and one diluted microemulsion for the protection of wood.

The physical and chemical stability were tested by a Collaborative International Pesticides Analytical Council (CIPAC) accelerated test: incubating the products for two weeks at 54 ?C. In both of the product-type formulations, problems occurred when attempting to obtain stable microemulsions incorporating both biocide molecules and extracts. After optimisation of the extraction procedures, formulations with better stability and synergic activity were obtained.

WP4: standardised biological and ecotoxicity laboratory tests

Wood protection

A large number of prototype formulations for the protection of wood have been prepared by Berkem SA. Several of those formulations with reduced fungicide content fulfil standard testing criteria against fungal degradation. The European standards followed were: testing against basidomycetic fungi (EN113) after aging by leaching (EN84) and evaporation (EN73), and resistance against blue staining fungi (EN152). The wood formulation prototype with insecticide still needs optimisation as it failed in termite testing (EN117 and EN118) after water leaching (EN84). However, these formulations passed the termite test after thermal evaporation (EN73) and also passed the test against Hylotropus larvae (EN46).

Urban pests

The insecticide sprayable prototypes were tested in knockdown assay against houseflies based on an Australian method in smaller glass cages. These water-based prototypes mainly failed in reproducing the synergic effect earlier found in the screenings in WP2. Only one formulation gave a slight synergic effect and could be used for the real case trials and could also be a starting point for further development.

Ecotoxicity

The ecotoxicity of two selected synergic extracts used in WP3 were tested on the following fresh-water organisms: fish, daphnia and algae.

The extract used for wood-protective formulations was found to be harmful to fish and toxic against daphnia and algae. However, the possibility of further reducing the content of toxic synthetic biocides in wood preservatives was evaluated in WP5.

The extract used to improve the knockdown effect of sprayable insecticide was evaluated as not toxic to the tested water organisms.

WP5: industrial validation

Semi-industrial evaluation of protective formulations for wood was performed by the impregnation of timber in an autoclave. It was found that the extract penetrated well and that the fungicide had a reasonable penetration up to 20 mm, which should be enough for wood intended for Class 3 use.

In the real case treatments against insect pests, it was confirmed that a surface application of one synergic insecticide prototype formulation was effective against cockroaches and also had a residual effect after one week.

A cost analysis was performed on the two types of prototype products. The addition of synergic extract to the wood protector would increase the price, although the reduced biocide content would be beneficial to the environment and to public opinion.

In the case of the sprayable insecticide it was evaluated that actually the price increase would be too great as a result of the addition of the synergic extract for the product to be commercially viable. Further development of the extraction method and starting material would be needed to solve this problem.

This WP has been defining the protection scheme for knowledge gained from the research, among the SMEs and regarding diffusion to wood-protection and urban-pest sectors. LThe SMEs are currently investigating the patentability of the discoveries and how to solve still-present technical problems, before having functional products to exploit on the market.

Potential impact

The potential impact of the results from the project is derived from the possibility to reduce the content of currently used biocides in the investigated products: wood protector and sprayable insecticide. The following impacts can be foreseen.

Environmental impact

1. The reduction of the use of conventional, more toxic biocide products, which are harmful to health and the environment.
2. The promotion of the use of treated wood and more environmentally friendly products in construction, civil works and infrastructure sectors as well as the agricultural sector, as opposed to materials treated with highly toxic biocide products or the use of alternative materials such as steel and concrete that are more energy-consuming and contribute greater CO2 emissions to the atmosphere.

The reduction of air, ground and underground water pollution caused by emissions being released into the atmosphere and the leaching of highly toxic biocide compounds.

Socio-economic and industrial impact

1. An increase in the added value of food and agriculture residues used for synergic extracts over others that are currently being used, which are less profitable and interesting.
2. The development of more economical and environmentally friendly biocidal products than are current available, by substantially reducing the concentration of toxin, and therefore:
(a) reducing the costs of treatment and offering more competitive biocidal products on the market;
(b)reducing costs and legal and environmental requirements in treatment of wood and in urban-disinfection processes.
3. The provision of new substances that may enable manufacturers of biocidal products to enter a new market for non-pollutant products that are guaranteed by standardised tests.
4. A boost to modernise and adapt the pest and wood industries to coming changes.

The foreseen use of the results and possible interested partner depend on which extract and type of product to be produced, as shown below.
1. patenting: any interested SMEs;
2. further development and optimisation: any interested SMEs and related research and development institutions;
3. Biocidal Products Directive (BPD) and Registration Evaluation Authorisation and Restriction of Chemicals (REACH) compliance: any interested SMEs;
4. manufacturing of extract or extracts: Rutland Biodynamics Ltd. and/or Berkem SA;
5. production of wood preservative and/or insect pesticide: DTS Oabe and/or Berkem SA and/or Linotech AB;
5. use of the final products: Quadrifolio, only in the Portuguese market.

Project website: for further news of the synergic pesticide products, their development and introduction on the market, please visit the webpage http://www.bimosyn.eu/ or contact the project co-ordinator, Ingemar.svensson@tecnalia.com.