Biopesticide-based products and strategies for control of tree pests. Professor Tariq M. Butt

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Biopesticide-based products and strategies for control of tree pests Professor Tariq M. Butt

Outline of Presentation Biopesticides Products and strategies for control of: Chestnut weevil and chestnut tortrix Pine processionary moth (PPM) Pine weevil (PW)

Biopesticides Biopesticides are naturally occurring plant protection products and include: Microbial biological control agents (fungi, viruses, bacteria) Semiochemicals (e.g. pheromones) Botanicals (plant extracts, essential oils) Global biopesticides market growing rapidly (CAGR 16%) - will reach US$7.6 billion by 2022.

Many drivers: Withdrawal of many pesticides (>67%, 2009) EU directives e.g. Directive 2009/128/EC-sustainable use of pesticides Supermarkets/consumers want zero residues Insecticide resistance in pest populations Environmental protection - promote biodiversity

Biopesticides weaknesses - slow acting, inconsistent results, narrow host range. It can take 2-7 days for entomopathogenic fungi (EPF) to kill speed depends on fungal virulence, dose and target host.

Biopesticides have their strengths - in some instances they are the best or only alternative to chemical pesticides. Efficacy enhanced using innovative strategies e.g. Lure & kill, Stress & kill

Metarhizium for control of chestnut pests Sweet chestnut (Castanea sativa) source of timber and nuts UK Chestnut production - 18,000 ha (Kent, Sussex) Nuts have high retail value

Vast sweet chestnut forests in Turkey. Turkey 2 nd largest producer in world ca. 64,750 tonnes (FAO Statistics, 2016) Trees and nuts attacked by various insect pests Growers do not want to use chemical pesticides impacts on health and biodiversity Chestnut weevil, (Curculio elephas) and Chestnut tortrix (Cydia splendana) lay eggs on nuts of oak and sweet chestnut

Curculio elephas Cydia splendana

Metarhizium best solution for pest control Fruit placed in pile on soil then covered with leaf litter. Heat generated causes fruit to dehisce and nuts fall out easy to harvest. Weevil and moth larvae leave nuts and pupate in soil. Pests are highly concentrated allows for targeted control.

Fruit (burrs) Metarhizium top-dress application Larvae pupate in top 5-10cm of soil. Highly concentrated easier to control. Asan, C., Hazir, S., Cimen, H., Ulug, D., Taylor, J., Butt, T. & Karagoz, M. (2017). An innovative strategy for control of the chestnut weevil Curculio elephas (Coleoptera: Curculionidae) using Metarhizium brunneum. Crop Protection 102, 147-153.

Larval mortality (%) Mean % mortality of Curculio elephas larvae following topdress application of M. brunneum strains V275 and 4556. Different lower case letters above the bars indicate significant differences at P<0.05. 100 80 a a Fewer adults emerge Outdoor to cause damage following season 60 40 b 20 0 Mb-4556 Mb-V275 Control Treatments

Mean larval mortality (%) Mean % mortality of Cydia splendana larvae by M. brunneum 4556 and the EPN Steinernema feltiae used alone or together. 100 80 ab b a Fewer adults emerge to cause damage following season 60 40 c 20 0 M. brunneum S. fel ae M. brunneum and S. fel ae Treatments Control

Pine processionary moth (PPM) Thaumetopoea pityocampa, T. wilkinsoni Invasive species spreading North from Mediterranean Major pest of pine feed on needles Stunt or kill trees Reduce pine nut yield

Urticating hairs pose a serious risk to humans and animals. Toxin (thaumetopoein) - can cause extreme skin irritation, respiratory problems, conjunctivitis, anaphylaxis (allergic reaction) and even blindness. Portero et al., 2012. Corneal Inflammation From Pine Processionary Caterpillar Hairs. Cornea, June 5 http://www.ncbi.nlm.nih.gov/pubmed/22673853 Battisti, A., Larsson, S., & Roques, A. (2017). Processionary moths and associated urtication risk: global change driven effects. Annual review of entomology, 62, 323-342. Costa, D., Esteban, J., Sanz, F., Vergara, J., & Huguet, E. (2016). Ocular lesions produced by pine processionary caterpillar setae (Thaumetopoea pityocampa) in dogs: a descriptive study. Veterinary ophthalmology, 19(6), 493-497.

Procession Pupation in soil Jan- Feb Mar- Apr May- June Nov- Dec Sept- Oct July- Aug Moth emergence Nests 3-5 th instar Urticating hairs Egg laying 1 st -2 nd instar

Biopesticide-based solutions for PPM control Solutions for all stages of development Mating disruption pheromone - targets adults, reduces oviposition Plant derived oviposition repellents Metarhizium strains kill eggs, larvae, pupae Botanicals kill larvae Aydin, T., Branco, M., Guven, O., Goncalves, H., Lima, A. Karaca, I., Butt, T.M. 2018. Significant mortality of eggs and young larvae of two Pine Processionary Moth species due to the entomopathogenic fungus Metarhizium brunneum. Biocontrol Science and Technology. 28, 317 331 Sönmez, E., Demir, İ., Bull, J., Butt, T. & Demirbağ, Z. (2017). Pine processionary moth (Thaumetopoea pityocampa, Lepidoptera: Thaumetopoeidae) larvae are highly susceptible to the entomopathogenic fungi Metarhizium brunneum and Beauveria bassiana. Biocontrol Science and Technology, 1-12

PPM eggs, larvae, pupae are susceptible to EPF PPM egg masses are conspicuous, accessible (i.e. easy to spray with EPF) Early instar (1 st, 2 nd ) larvae have no urticating hairs no risk to operators

M. brunneum (V275/ARSEF4556) conidia adhere to surface of egg mass Reduces egg hatch and kills emergent larvae (>90%) 98-100% mortality of 2 nd instars achieved 2-4 days pi, using dose 1 X 10 6 conidia ml -1 Aydin, T., Branco, M., Guven, O., Goncalves, H., Lima, A. Karaca, I., Butt, T.M. 2018. Significant mortality of eggs and young larvae of two Pine Processionary Moth species due to the entomopathogenic fungus Metarhizium brunneum. Biocontrol Science and Technology. 28, 317 331

Older (3 rd -5 th ) instar larvae are also susceptible to EPF Excellent control when EPF injected into nest: 100% dead 3-6 days pi, using dose 1 X 10 8 conidia ml -1. Larvae rest inside nest during day and feed at night. No risk to operator. Good horizontal transfer of conidia from treated to untreated larvae inside the nest. Sönmez, E., Demir, İ., Bull, J., Butt, T. & Demirbağ, Z. (2017). Pine processionary moth (Thaumetopoea pityocampa, Lepidoptera: Thaumetopoeidae) larvae are highly susceptible to the entomopathogenic fungi Metarhizium brunneum and Beauveria bassiana. Biocontrol Science and Technology, 1-12

Excellent results obtained both in lab and field trials when 10ml conidial suspension injected inside nest. Mean % mortality of 3 M. brunneum instar PPM strain Lab Field V275 100 86.9 ± 6.4 ARSEF 4556 100 -- Control 0.8 ± 0.4 3.7 ± 1.6

High mortality of last instar larvae and pupae Pre-treating pupation soil with M. brunneum (V275/ARSEF4556) gives good control 70-90% mortality using dose of 1.4 X 10 11 conidia kg -1 soil 20-40% die as larvae remainder die inside pupal cell Inoculum available to infect larvae following year

Botanicals offer alternative to EPF Large number of botanicals (plant extracts, essential oils) available Most botanicals used in cosmetic sector considered safe Not all botanicals have insecticidal properties.

Mortality of PPM larvae 7 days pi following injection of nest with 10ml 1% aq. v/v plant oil. Treatment Mean % mortality L3-L4 Ginger* Eucalyptus Rosemary Control Laboratory 71.2 ± 8.5 24.2 ± 6.7 30.0 ± 4.4 1.3 ± 0.8 Field 87.4 ± 6.7 86.7 ± 4.6 76.1 ± 5.5 2.3± 0.9 100% mortality, faster control using 10% v/v Larvae stop feeding (less frass), stunted growth, stressed, eventually die. Higher mortality in field than lab. botanical control

EPF and essential plant oils could be used to control oak processionary moth (OPM, Thaumetopoea processionea) OPM often forms nest on tree trunk easy to treat with EPF/botanical Biopesticides safer than chemical (deltamethrin) alternative. Forest Commission

Semiochemicals Behaviour modifying chemicals include pheromones and kairomones. Species specific sex pheromones widely used for: Pest monitoring Pest control e.g. mating disruption, mass trapping

Pests use plant volatiles (kairomones) to: locate suitable hosts for food or oviposition avoid unsuitable hosts Attractant compounds could be used for pest monitoring mass trapping lure & kill pest control

Benefit of Lure & Kill pest control strategy Attractants lure pests to control agent. Targeted control (reduces area to be treated). Reduces application rates and cost of control. Useful in environmentally sensitive or difficult terrain

Progress to date

PW highly damaging to saplings PW lay eggs on freshly cut stumps Attractants encourage oviposition on treated stumps Attractants infused in polyvinyl acetate string More PW larvae recovered from treated than control plots.

EPF-EPN combinations give >90% control. Encouraging oviposition on selected stumps allows for targeted control reducing application rates and reducing costs. Fewer adults emerge to cause damage to saplings

Total number per cell Billets - lure Billets + lure Lure & Kill for PW control Billets release volatiles that attract adult PW. They also act as feeding station. Several billets are more attractive than single billet. Number of billets reduced using lures in biodegradable dispensers.

Ultimate goal is to place EPF under billet with lure as part of Lure & Kill strategy for pine weevil control. Field trials - encouraging results. 14 12 10 8 % Mortality of PW collected at 3 time points (billet + lure + EPF) 6 4 2 0 week 1 week 2 week 3

Summary EPF, plant oils and kairomones have been identified for the control of PPM, PW and other tree pests. These biopesticides offer an environmentally friendly alternative to conventional chemical pesticides. EPF kill both adult and juvenile stages of most pest species

Botanicals show much promise for the control of PPM larvae without exposing operators to the urticating hairs. Semiochemicals will play an increasingly important role in pest control programmes. Attractants could be used to lure pests to control agents reducing the area to be treated and help reduce cost.

Some of the products and strategies could be used to control other pest species that threaten tree and human health. Oils and EPF developed for PPM could be used to control larvae of OPM

Research partners (UK) Acknowledgements Industry partners Research partners (Turkey, Portugal) Prof Manuela Branco, UoL Dr Selcuk Hazir, Dr Mehmet Karagoz, AMU Prof Ismail Karaca & Prof Ozlem Guven, SDMU Prof Zhini Demirbag & Prof Ismail Demir, KTU