Varroa Sensitive Hygiene-VSH

Selection of Varroa Sensitive Hygiene (VSH) Behaviour in Honey Bees

Stéphanie Rouleau-Breton, MSc : phenotype study Marie Lou Morin, MSc: phenotype study Supervisor : prof Pierre Giovenazzo Laurence Auger MSc candidate: genotype study Supervisor : prof Nicolas Derome 1 University Laval and Centre de recherche en sciences animales de Deschambault

2 Beekeeping in Canada

Honey bee imports Importance of canadian honey bee breeding 300000

250000

200000

150000 3 100000

50000

8 9 0 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 6 7 8 9 0 1 2 3 4 5 6

8 8 9 9 9 9 9 9 9 9 9 9 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1

9 9 9 9 9 9 9 9 9 9 9 9 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

1 1 1 1 1 1 1 1 1 1 1 1 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2

Queens Packages

Source: Stephen Page, Sector specialist, Horticulture Division AAFC Sanitary security vs imports

• Risks: • Undesired pathogens • Misadapted and undesired genotypes (Büchler et al. 2014, Parker et al. 2010)

http://aratakihoneyrotorua.co.nz

https://www.mielsdanicet.com

4 Sanitary risk assessment

• Canadian food inspection agency, 2013 Risk Assessment on the Importation of Honey Bee (Apis mellifera) Packages from the United States of America.

5 Our objective • Establish a long term breeding program • CRSAD • Extension • Evaluation of genetic stock, Selection selection and breeding.

• Local Queen breeders Multiplication • Performance evaluation • Multiplication and distribution of selected lines. Production • Commercial beekeepers • Performance evaluation

6 Cost of maintaining a selection program

A breeding program is not just multiplying queens!

Management Winter hardiness Hygienic behaviour

Honey production Varroa tolerance: VSH/SMR

Who should pay for all this work???? • Beekeeper: Queen price = ? • Government/University support Instrumental insemination • Is it worth it? 7 CRSAD honey bee selection program Integration of VSH selection criteria

Start 2010 • 50 queens Buckfast, 5 lines (Keld Branstrup from Denmark) • 20 queens VSH, 2 lines Glen Apiaries USA • 60 Québec local stock, 3 lines Each following year 12 to 15 new lines = 100 to 140 colonies / year • Selection method: criteria priority • Wintering success (survival and spring strenght) • Defensive behaviour (never reproduce an agressive colony) • Hygienic behaviour (nitrogen freeze) • Honey production (kg) • Varroa population build up (varroa drop) • 2015 : new 5 breeder VSH queens (Happy Bee apiaries USA) • 2016: Start BeeOmics project • 2017 Segolène Maucourt PhD : heritability and breeding values

8 Notre approche: LaVSH sélection behaviour d’un comportement

Honey bee development and varroa infestation Honey bee with VSH behaviour

Larvae Egg laying

Larval nutrition Worker uncapping cell Varroa enters cell Worker removing parasitized larvae Worker capping cell

Founding mother

Varroa reproduction Adult mâle Egg

Protonymph

Young bee emerges Deutonymph 9

https://aristabeeresearch.org/varroa-resistance/ Harbo et Harris, 2005; Boecking et Spivak, 1999; Evans et Spivak, 2010 Varroa development

Adult female Female deutonymphe

Protonymph Larvae

Adult mâle Mâle deutonymph

Cycle de vie de Varroa destructor; quatre photos de Gilles San Martin/Flickr; deux photos de MrJEberhardt/Youtube. 10 Rosenkranz et al. 2010 VSH observations

o No viable descendant fertile females o At 9 days post capping: o no female deutonymph or no male o Suppressed mite reproduction-SMR o Varroa sensitive hygiene-VSH

3. Infested frames placed in receiving 1. Infestation of colony to be tested frames in varroa for VSH donor colonies 4. Evaluation of reproductive succes 2. Evaluation of initial of varroa and final varroa infestation varroa intestation level level

12 Büchler et al. (2014) VSH testing

1. Excluder frame with queen in tested colony

A B

http://apiexpert.eu/shop/ ©Andrée Rousseau

Leave 48h for queen laying uniformity

13 VSH testing

2. Infestation of frame in varroa doner colonies (with at least 10% infestation rates) + D C

©Andrée Rousseau https://www.honeyflow.com/resources/blog/beginner-beekeeping-with-a-flow-hive/p/200

Wait 10 à 11 days for capping and varroa invasion14 VSH testing

3. Évaluation of initial infestation rates F + E

http://www.resistantbees.com/hyg_e.html

https://www.honeyflow.com/resources/blog/beginner-beekeeping-with-a-flow-hive/p/200

https://psmag.com/environment/bees-bred-with-some-special-genes-are-killing-it 15 X 75 cells / frame side VSH testing

4. Placing infested frame in receiving colony Donor colony Receiving colony

VSH behavior testing for at least 7 days

16 L’emplacement des VSHruchers testing expérimentaux

4. Apiaries with receiving colonies to be tested for VSH

N = 20

Apiary 1

N = 20

Apiary 2

17 VSH testing

Receiving colony

VSH behavior testing for at least 7 days

18 VSH Project

5. Evaluation of final varroa intestation level

X 75 cells / frame side 19 VSH testing

6. Evaluation of varroa reproductive success and recapping activity

Minimum 30 cells with one mother varroa

20 VSH testing

Reproductive success = reproductive mother varroa X 100 N of infected cells

Suppressed mite reproduction-SMR = non reproductive mother varroa X 100 N of infected cells

Varroa sensitive hygiene -VSH = % Final varroa infest - % Initial varroa infest

http://www.omafra.gov.on.ca/french/food/inspection/bees/varroamite.htm

21 VSH testing

22 2016 VSH preliminary results

Table. Mite reproduction for 10 colonies in 2016 . Varroa mites were considered reproductive when at least one female deutonymph and one male were present in the cell and as non-reproductive if one of the two was absent. N cells with 1 Colony # Genetic Line Rep varroa Non rep varroa Fertility of mites (%) Cell manipulation (%) varroa 28 2 26 7,1 50,0 233 14-9-dm 30 7 23 23,3 74,1 30 3 27 11,1 32,4 305 14-8-cfhm 30 2 28 6,7 10,9 30 4 26 13,3 20,0 398 14-6-h 30 1 30 3,3 26,0 31 1 30 3,2 82,5 340 14-12-ch 30 5 25 16,7 5,5 24 11 14 45,8 20,0 317 14-6-h 30 4 26 13,3 26,3 32 11 21 34,4 17,0 604 15-v-1 28 4 24 14,3 33,0 30 2 28 6,7 60,7 329 (VSH) VSH-Carniolian 30 6 24 20,0 19,6 30 9 21 30,0 41,0 347 (VSH) VSH-Carniolian 30 11 19 40,0 76,8 11 3 8 27,3 0,0 333 (VSH) VSH POL-Line 21 8 13 38,1 10,0 23 31 13 18 41,9 28,0 282 (VSH) VSH-Carniolian 2017 SMR preliminary results

• 4 beekeepers, 20 colonies per 12

beekeeper 10 • 1 brood frame sampled and frozen 8 • 2 first weeks of August 2017 6 • Varroa infestation rates 5%-10% • Maximum 600 cells verified /only the 4

ones with 30 single infested cells are 2

presented in figure cells infested with30 frames : colonies Nb • 2 to 4 hours per colony! 0

%SMR

24 VSH Ontario-Québec project 2018-2021

• Collaboration with OBA tech transfer team

• $200 000 = New Directions Research Program, OMAFRA • $66 000 = University Laval • $98 000 = Centre de recherche en sciences animales de Descahmbault

25 VSH Ontario-Québec project 2018-2021

• Identify VSH colonies within the CRSAD breeder colonies • Performance assessment of the CRSAD-VSH selected lineages, • Determine the heritability of the VSH trait, • Evaluate the relation of the VSH trait with other selection criteria, • Evaluate if there is an ideal percentage of expression of the VSH/SMR behavioral trait that that should be targeted so that colonies resist the mites while expressing the attributes of the other selection criteria effectively. • Distribute VSH lineages to breeders in Ontario (2x) and Québec (2x) and evaluate their performances in situ. • Participating breeders will multiply and offer VSH lineages to their beekeeper customers across Canada.

26 VSH relation with other selection criteria

Winter Winter consumptio survival n

Varroa Spring infestation population

Defensive Swarming Honey behaviour behaviour production

Hygienic Summer behaviour population SMR

27 http://www.sciencealert.com/we-really-might-be-killing-our-bees-according-to-new-evidence? VSH relation with other selection criteria

Spring buildup

Devensive behaviour

https://imgur.com/gallery/gsQaZ

= Stings

Hygienic behaviour

= % removed 24h

Varroa is still a major beekeeping problem

VSH and varroa fertility studies are very promising

Good collaboration Ontario – Québec