Project leader: 
Estatuto projecto: 

Novas estratégias ecológicas para o biocontrolo do vector do nemátodo da madeira do pinheiro na Eurásia

Acronym: EcoVector

Reference: PTDC/AGR-TEC/3049/2014

Funding: FCT, PT2020, FEDER

Duration: 01/07/2016 to 01/05/2020

Principal Researcher: Mara E. M. Braga

Proponent Institution: PRODEQ - Associação para o Desenvolvimento da Engenharia Química (PRODEQ)

Other institutions: Universidade de Coimbra (UC) and INIAV

Start date: 01-07-2016

Final date: 01-05-2020

Financing granted: € 165.624,00


Project overview

The pinewood nematode (PWN), Bursaphelenchus xylophilus, is the causal agent of pine wilt disease (PWD) which is causing great losses in coniferous forests worldwide. Pinus trees are the main hosts for the PWN, and transmission from one host tree to another requires an insect vector. Cerambycids of the genus Monochamus is considered to be the most important vectors, being M. galloprovincialis the only vector in Portugal. Besides the ecological impacts associated to pine forest destruction, the PWN is being responsible for enormous economic impacts in Portugal and, in the future, it can become a real threat to all Eurasia and North America. These economic impacts are mainly related to timber losses, to timber/wood products trade restrictions/reduction and the associated costs to additional phytosanitary procedures and PWN control actions.

This multidisciplinary project aims to develop new and more efficient phytochemical-based formulations for the control of the PWN insect vector. Phytochemicals can be alternative active ingredients (AIs) to pesticides because they are usually safer for the environment and human health than traditional synthetic ones. A large number of different plant secondary metabolites (such as essential oils, phenolics, quinones, tannins, etc) were already tested as efficient attractants/repellents for the PWN vector. Nevertheless, further research is still required to obtain formulations containing those attracting or repelling phytochemicals with proved efficacy on the field, prolonged action, easy to apply and with a reasonable cost.  

Formulations based on biodegradable polymer monoliths and/or particles, loaded with specific and selective volatile AIs or mixtures of them were developed. Chosen biodegradable polymers such as poly(caprolactone) (PCL) and their biocomposites, as well as chitosan were used. The incorporation of AIs into these types of biopolymer monoliths/particles allowed their controlled, prolonged and effective release.

Two different product concepts were explored: (i) commercially available trap lures, to be placed in the forest, containing prolonged attractive-releasing monoliths and/or particles; (ii) prolonged repellent-releasing particles to be spread over timber or selected areas. Polymer monoliths were produced by supercritical carbon dioxide mixing/foaming, while particles were prepared by solvent evaporation from o/w emulsions. Both techniques are advantageous since they allow materials processing at relatively low temperatures, thus avoiding the degradation of any involved thermally labile AIs, and promoting higher AIs loading efficiencies. Total/relative AIs composition, biopolymers (and biopolymer molecular weights) and process conditions varied to obtain monoliths/particles with different physical and degradation/release properties and, ultimately, to optimize the final product performance. All obtained materials were chemically and physically characterized by a wide range of analytical techniques. Experimental determination and computer modelling of the AIs controlled release (overtime) was carried out and correlated with formulation variables and processing conditions. 

Chemical Product Engineering (CPE) concepts and methodologies provided the unifying line of the project, promoting the definition of adequate product performance metrics and respective target values (necessarily including the reduction in vector prevalence), which guided the detailed definition of the experimental tests that were performed. Also, a CPE approach implies an integrated perspective of product/process development and associated management of information, tests and models involved. To the best of our knowledge, this project will be the first effort, and a systematic one, to prepare biodegradable and natural-origin controlled/prolonged release systems of volatile attractant/repellent AIs for the control of the PWN insect vector.

Project outcomes

Scientific articles

1. M.C. Gaspar, B. Agostinho, L. Fonseca, I. Abrantes, H.C. de Sousa, M.E.M. Braga. Impact of the pinewood nematode on naturally-emitted volatiles and scCO2 extracts from Pinus pinaster branches: a comparison with P. pinea. J. of Supercritical Fluids 159 (2020) 104784.

2. M.C. Gaspar, H.C. de Sousa, I.J. Seabra, M.E.M. Braga. Environmentally-safe scCO2 P. pinaster branches extracts: Composition and properties. Journal of CO2 Utilization, 37 (2020) 74-84.

Master Thesis

  1. Vanessa Monteiro Leal, Development of formulations based on attractants/repellents to control the insect vector of pinewood nematode (PWN). 

Departamento de Engenharia Química – FCT – Universidade de Coimbra, sob supervisão de Mara E. M. Braga e Hermínio de Sousa. Defesa 27 de Setembro de 2018;   

  1. Andreia Catarina Ribeiro Madeira, Incorporation of volatile repellent compounds for the development of formulations to control insects’ incident to agroforestry ecosystems. Departamento de Engenharia Química – FCT – Universidade de Coimbra, sob supervisão de Mara E. M. Braga e Ana Dias. Defesa em Fevereiro de 2019;

Summer internship

  1. Diana Gonçalves – Quemical Engineering, program “Estágio de Verão” - Universidade de Coimbra, 2019.
  2. João Coutinho – Biomedical Engineering, program “Estágio de Verão” - Universidade de Coimbra, 2019.
  3. Ana Rita Fernandes – Pharmaceutical Sciences, program “Estágio de Verão” – Universidade de Coimbra, 2019.
  4. Beatriz Agostinho – Chemical Engineering, program “Estágio de Verão” – Universidade de Coimbra, 2018.
  5. Vanessa Leal – Chemical Engineering, program “Estágio de Verão” – Universidade de Coimbra, 2017.

Communications in International Conferences

1) M.C. Gaspar, B. Agostinho, J. Leocádio, L. Fonseca, I. Abrantes, Hermínio C. De Sousa, M. E. M. Braga. Pinus pinaster and P. pinea volatile compounds: their possible action on pinewood nematode and its insect vector. International Symposium IUFRO on Pine Wilt Disease, Orléans (France), 9-13 March 2020, cancelado devido à pandemia COVID19.

2) Fernando P. Bernardo, Mara E. M. Braga, Rita Chim. A wind tunnel method to develop products for controlled delivery of volatiles: experimental apparatus and mathematical model. 12th European Congress of Chemical Engineering (ECCE 12), Florence, Italy, 15-19 Sep 2019.

3) M.E.M. Braga, Porous matrices based-formulations for improvement of volatiles loading/release for insect control. V Iberoamerican Conference on Supercritical Fluids. September 02-06, 2019. Campinas, Brazil.

4) M. C. Gaspar, B. Agostinho, L. Fonseca, I. Abrantes, H. C. de Sousa, M. E. M. Braga. Pinewood nematode and its impact on volatiles and scCO2 extracts from Pinus pinaster branches. 17th European Meeting on Supercritical Fluids (EMSF 2019) – 7th European Meeting on High Pressure Technology. Ciudad Real, Spain 8-11 Apr 2019.

5) M. C. Gaspar, H. C. de Sousa, I. J. Seabra, M. E. M. Braga. Extracting the natural insecticide abietadiene from P. pinaster branches using scCO2. 12th International Symposium on Supercritical Fluids (12th ISSF 2018), Antibes-Juan-Les-Pins, France, 22-25 April 2018

6) M.E.M. Braga, R.Chim, F.P. Bernardo, H.C. de Sousa. Active ingredient release from porous matrices to control the insect vector of pine wilt disease. 12th International Symposium on Supercritical Fluids (12th ISSF 2018), Antibes-Juan-Les-Pins, France, 22-25 April 2018.

7) R.Chim, H.C. de Sousa, F.P.M. Bernardo, M.E.M. Braga. Dispersão do α-pineno em matrizes porosas para o controlo do insecto vector (Monochamus spp.) do nemátode do pinheiro. II Congresso Nacional das Escolas Superiores Agrárias (II CNESA), Escola Superior Agrária de Bragança, Novembro 16-18, 2017, Elvas, Portugal. 

8) L. Fonseca, C.M.N. Maleita, I. Esteves, M.E.M. Braga, I. Abrantes, H. C. de Sousa. Naphtoquinones: bionematicides for pinewood nematode eradication from wood. International Union of Forest Research Organizations (IUFRO 2017), September 18-22, 2017, Freiburg, Germany.

9) J. Leocádio, M.C. Gaspar, F. Bernardo, P. Naves, E. Sousa, H.C. de Sousa, M.E.M. Braga. Searching a way of controlling the Pine Wood Nematode insect vector using PCL based matrices obtained by supercritical CO2 foaming. 1º Encontro Ibérico de Fluidos Supercríticos (EIFS 2020), Santiago de Compostela, Espanha, 18-19 de Fevereiro de 2020.

10) V. M. Leal, M. C. Gaspar, H. C. de Sousa, M. E. M. Braga. Porous matrices based-formulations for improvement of α-pinene loading and release. 17th European Meeting on Supercritical Fluids (EMSF 2019) – 7th European Meeting on High Pressure Technology. 8-11 Apr 2019, Ciudad Real, Spain.

11) P. Naves, C. Trindade, M.E.M. Braga, M.C. Gaspar, E. Sousa. Is host selection of Mediterranean pines by Monochamus galloprovincialis (Coleoptera; Cerambycidae) related with Pine Wilt Disease incidence? XI European Congress of Entomology, 2-6 July 2018, Naples- Italy.

Team project

Mara Elga M. Braga – PRODEQ - CIEPQPF/FCTUC           

Hermínio C. de Sousa – PRODEQ - CIEPQPF/FCTUC                   

Marisa C. Gaspar – PRODEQ - CIEPQPF/FCTUC               

Fernando Pedro Martins Bernardo – PRODEQ - CIEPQPF/FCTUC            

Ana Antunes Dias – PRODEQ - CIEPQPF/FCTUC              

Maria da Graça Rasteiro – PRODEQ - CIEPQPF/FCTUC   

Inês Seabra – PRODEQ - CIEPQPF/FCTUC            

João Leocádio – PRODEQ - CIEPQPF/FCTUC       


Isabel Abrantes – CEF – IATV/FCTUC

Luís Fonseca – CEF – IATV/FCTUC

Isabel Luci Pisa Mata da Conceição – CEF – IATV/FCTUC

Edmundo Sousa - INIAV

Pedro Naves - INIAV