Krispn Given, Purdue University

https://extension.entm.purdue.edu/beehive/

How to Breed for Behavioral Resistance to Varroa Destructor

• Increase honey Production• Improve overwintering ability• Decrease defensive behavior• Reduce swarming• Reduce or increase use of propolis• Change color• Decrease incidence of diseases• Improve defense against varroa destructor*• Other personal reasons

Why breed bees?

Breeding versus queen rearing

Most important part of a breedingprogram

• Decide on a trait (phenotype)• Develop a assay• Measure the foundation population• Select parents• Control mating using a design• Evaluate, document, and verify• Select parents

Bee Genetics + Breeding

Mendel’s bee house

16 CHROMOSOMES

~260 million bases (MbLOW REPETIVE DNA

HAPLOID MALES

HIGHRECOMBINATIONRATES!

Honey Bees

CHROMOSOMES

– Composed of coiled, twisted DNA and coatedwith proteins

– Each species has aspecific number

– They are organized inpairs

(one comes from the motherand the other from thefather)

Genetic gender and caste

• Queens lay two types of eggs

• Males develop from unfertilized eggs

• Females develop from fertilized eggs

HONEY BEE GENETICSBees have 16 paired chromosomes.

Haplodiploidy• Males honey bees have one of each chromosome (haploid 16)• Females have two of each chromosome (diploid 32)

Polyandry•The queen mates with 10 to 15 males, resulting in many sub-families within the hive’s family.

Many important behaviors are influenced by genes!•stinging•pollen foraging•undertaking

•hygienic behavior•brood rearing traits•swarming tendency

•propolis collection•honey production•even learning!

XX

FEMALES

X

crossing-over

WHAT HAPPENS WHEN YOU CROSSTWO DIFFERENT STOCKS OF BEES

gives you

X

HYBRID CROSS

or

HYBRIDSHIGH LINE LOW LINE

35%

l.5%

5%

0%

Tro t a US manag 1ed honey be 1e colonies Loss EstimatesD A c c e p t a b l·e W in te lf Loss D T1ola l Winte r Loss T otail Annual loss1

' - '......-

-=- ! - •...... _

, ..............- - · ,_,..... . -- -

- - i - --

' -, _ ......

- ! - - = -

-........... - >-

· - - - -_ I."

_ _ IJ · •" • ·- ---

-

rr::&---1 -=-

_:5'Mt: 7 ,_ •

· - ,.....-

--

-I

--

-I

i-I

I -

73291- 1 API LIFE VAR Thymo l (74.09%), Oi l of eucalyptus (16%), Menthol (3.73%)

Formic acid (46.7%)

Thymo l (2 5%)

75710 - 2

796 7 1- 1MITE-AWAY QUICK STRIPS

APIGUARD

9 12 6 6 - 1 OXALIC ACID DIHYDRAT E

OXALICACID D HYDRATE

OXALICACID D HYDRATE

CHECKMITE+ BEE H IVE PESTCONTROL STRIP

CHECKM TE+ BEE HIVE PESTCONTROL STRIP

Oxali c acid (100%)

91266-1-73291

91266-1-91832

1155 6 - 138 Coumaphos (10%)

11556-138-61671

Registration #

2724-406

Product Name

ZOECON RF-318 APISTAN STRIPActive Ingredient

Fluvalinate (10.25%)2724-406-62042 AP STAN ANTI-VARROA MITE STR PS

61671-3 FOR-MITE Formic acid (65.9%)

70950-2 AVACHEM SUCROSE OCTANOATE [40.0%)

Sucrose octanoate (40%)

70950-2-2205 SUCROCIDE

70950-2-84710 SUCRASHIELD

83623-2 HOPGUARD II Hop beta acids resin (16%)

87243-1 Apivar Amitraz (3.33%)

Adult female invades cell before its sealed

Dr. Harry Laidlaw

female immerses herself in brood food, lays first egg after about 72 hours.

First egg laid is a haploid male, then mites mate in cell and must mature before the bee emerges (usually about 4 females emerge).

Varroa mites prefer drone brood but also invade worker cells. Most mites (80%) will be in the sealed drone and worker brood

THEY CAME FROM ASIAN HONEY BEES

In Apis cerana Varroa only reproduces in dronebrood

30k

Cell ofBro'od

Mite

\ No.

\ /\ /

\

I" I

I

>\I \/ I "'

v /

Jan Feb Mar

/----- r - - _ - - 0

Varroa Population Dynamics 3k40k

2.5k

2k

1.5k20k

1k

10k0.5k

0Apl May Jun Jul Aug Sep Oct Nov Dec

PARASITIC MITE SYNDROME

DWV

Virus and brood diseases appear

• Acute bee paralysis (ABPV)• Israel acute bee virus (IAPV)• Kashmir bee virus (KBV)• Black queen cell virus (BQCV)• Deformed wing virus (DWV)

• 22 known viruses!

some vectored viruses from varroa

Monitoring Mites in YourHive

• Uncapping drone brood

• Ether Roll• Ethanol wash• Sugar Roll• Sticky Boards

Looking for mites in drone brood

Looking for mites on adults

WHAT TRAIT WILL YOUMEASURE AND HOW TO

MEASURE IT?

1)How do you choose the parents? (needto evaluate and keep good records)

2)how do you control mating's?(need to do I.I. or have drone sourceflooding in your mating yard)

PRACTICAL WAY TO SELECT FOR VARROA RESISTANCE:

GRAFT FROM STRONG HIVES THAT DON’T GROW MITES

You need try to control mating's

Try to have good colonies with drones in the mating yard.

You need a trait you can evaluate, something that is beneficial,and that is heritable.

Start with bees that have good traits for your area!

1

50.00%

25.00%

12.50% 11.25%

5.63%2.81% 1.41% 0.70% 0.35% 0.18%

0

0.1

0.2

0.3

0.4

0.5

0.8

Loss of introduced genes open mating for 10 generations.0.7

0.6

0.9

1

1 2 3 4 5 6 7 8 9 10

PROBLEM:

Hygienic Behavior is Heritable(W. Rothenbuler 1964)

100%

0

Generations of Selection(made up data)

% D

EA

DB

RO

OD

RE

MO

VE

D

50%

Another example:Response to Selection for Pollen Hoarding

USDA Baton Rouge bee lab

VSH BEHAVIOR

Greg Hunt

University of Warwick

© Rodger Dewhurst 2006 and 2008

www.ces.ncsu.edu

•Varroa-sensitivehygienic behavior(VSH)

•Detection and removal of varroa infested brood

Making uniform colonies

VARROA MITES INSIDE THE CELL

Good age to check for reproductive mites

VSH is an indirect attack on Varroa

extension.org/bee_health

Measure changes in:capped cel l

uncapped

chew, remove / '

% uncapped% chewed% recapped

% varroa infestation% reproductive varroa% varroa with no progeny

recap

Figure 1. - Types of changes in brood combs that can be measured without and with a stereomicroscope. For example, the percentage of pupae uncapped during a certain period of t me couldpotentially be used to breed for varroa resistance. Uncapped pupae are visible without a microscope, but measuring the % uncapp ed pupae is highly var able an d does not always predict VSH behavior. Onthe other hand, measuring the percentage ofinfertile mites (those without progeny) predicts strongvarroa resistance, butit requires detailed microscopic examinations of brood to measure.

Visible w/omagnification.but imprecise

Precise, but requiremagnification

3 prepupa

4 fi·rst,protonymphe(ma1l1e)

pupa,white 1eyes

Days pastcapping

Brood stage Eldest offspr inginnormally 11eproducing

mites

7-9 pupalpurp1eeyes

first femaredeu1onymph

10-11 first adultdaughter

pupat,brown headl. blackeyes

5-6 pupa, second proto-pnk eyes ny1mph

(female}

Uncapping of mite-infested cells due to VSH

24hrs LATER!

GENERAL HYGIENIC BEHAVIOR

Freezing capped brood with liquid nitrogen

(250 ml)

OR PIN PRICK METHOD

Greg Hunt

GROOMING BEHAVIOR

MIGUEL AND ERNESTO’S STUDYIn Mexico

Found that grooming behavior was important

Those with the lowest infestation overall had low adult infestation show highest mite drop !

They also had the most chewed mites on board, and also in the lab assay.

% Mites Chewed in Hives60

50

40

30

20

10

0Africanized/ European

High/Low Gen. 1

Russians/ Unselected

High/Low Gen. 2

% Mites Removed in Assay30

25

20

15

10

5

0Africanized/ European

High/Low Gen. 1

Russians/ Unselected

High/Low Gen. 2

What is responsible for behavioral traits?

• Genotype of queens and workers

• Environmental factors

Grooming Behavior

MITES IN BROOD

50

40

30

20

10

0

%IN

FEST

ATIO

N

LOW GROUPHIGH GROUP

2 3 4MONTH

5 6

MITES ON ADULTS

35302520151050

%IN

FEST

ATIO

N

LOW GROUPHIGH GROUP

2 3 4MONTH

5 6

Selec t ing fo r c hew ed mite s iss t ic ky business !

'

H l l i f il .>, , i:•:r 1r 1I I I l il i f i lf #I°! ·l l i l I i i I i I i i !11 1· - f . • 1 1 Lill:-.a._,.....1.L. L. : ii. . . . . . . .J. . . r r .-Y ju ; 1I ' .. . _

/.!i £ SK Olo Bf f /

_.... ---

:•,I JI ,: •

.• 't : .• -

• ' I fI I • -

i I I i ..:I j• -. I

f2I If I

? 3 JO?; to••

r r r ' ' t 1

Current Selection Program: Grooming and Biting Behavior

Hierarchical Selection:

Measure proportion chewed mites.

Eliminate those that show disease or did not control mite population growth.

Eliminate those that do not pass freeze-killed hygienic test.

Select breeder colonies.

• Started with SURVIVORS, VSH + Russians• Method for the first 7 years:• Measure mite drop (and strength) multiple

times• Measure honey yield• Test best sources for (VSH) hygienic

behavior• Use instrumental insemination.

Response to selection at the colony level is also at the individual

Initiating grooming dance* Biting

UndertakerVSH

grooming

General hygiene

Instrumental Insemination

High x high chewers are crossed and monitored For more response to selection

Daughter queens are then grafted from these*

Collecting and counting chewedmites

(p = 0.028)

Photo ark

Mites t h a t h a v e b e e n c h e w e d b y t h e m ite-b i t ers

7 2 ? 3 ID 5 t oI I f

: I ! ! -t : I :: t I te. • . .

• I • •• • 1" •

..t

I •• J I i ..., '- .:-

, , , , , ,. ,, ' \Jf \ ! I , ! \ c

J 1 1 · , , I- I I iI f \ I 11 I I... . .i r c Jf f 11 I I• I I • --J j I fl i J. I Jf

, . ( j ! 1 j

I f I '- - - - --\-- II

•

..- '_...'__._.......,....g- - - ---·....- -.-·---\ , --··------

- ·-··-··-'·... ... ' . ' '..........._ ----..' - -···--. ._·--- -- -- - ···· , ..........---- -:::r--- ·-- -- - ·- l!I"- : - - - -

........ ...-. -'...' . '---··er : - . -

•- - -·---..--- - "'.. ........... 0..... -· -, . , I II

. . ' .::r-.:::::-=-=·=----- - ·-·-·-- ......:::

-·----1-•--• --• --- - - - - - - t.n-·-·- ·-- -- ..1:::' -- - -. ; - -----.....---- --- ·- - ..._--I I ' I ' I

- - - ·- - - - ...c::- .....___...... . . _ _ _ _ ()CJ..._ ___I I I I

i f l ! i l i l i ! ! i·: i i' j;;I i i !

·1:J i'i l i l°1I i !i-:tJc"!.l t f l f i l l I iI ' l 1· -f H ; : • I · • ·• 1

,, , : 1 1i 1 !· + • '""'" H ..., . . , . I 1

/.S £S sg Ob Bf

-------·----··-.....--'·---.·--.-_-._-...·. ----......·-.- ..- --· - -- -·'

.I i i l_l r : l _I ;: j { lj I l li f !1 I i i i l H - ! ; ( J l ll If I I Ii I ! i i -fl l i ·11111H 111i iH ·n n ·1n!f l l I l l ! fi i l ) l ! j' llJ

?Z: IJ, 11 , , dZ

----- t".".l"t"-_-_- -_...._ _

--

. . . .....c..:,. .·_ -. .--.........._.--,, ,·-·- ------------ ""

• • • • -- - -• • I•

--·- - -_-..-·--

2014 August P<0.001

50

45

40

35

30

25

20

15

10

5

02006 2007 2008 2009 2010 2011

YEAR2012 2013 2014 2015

PE

RC

EN

TC

HE

WE

DMITE-BITING IS A HERITABLE TRAIT.

The bite of the bee

contains 2-heptanone

Varroa

Wax moth larva

n

How to select for bees that bite varroa mites and groom them from their bodiesKrispn J. Given, Greg J. Hunt, David M. Shenefield, Ginger D. Davidson Dwight C. Wells

and Dan P. O’Hanlon

Abstract

Grooming behavior is one of the key mechanism honey bees (Apis Melifera) execute to kill and remove Varroa destructor from their bodies. Two of the grooming behaviors identified with chewing and mite removal are responsible. Through selection with instrumental insemination we have seen Varroa Destructor chewing levels go from 3% to 43%. The

Heartland Honey Bee Breeders Cooperative was initiated to promote the production and dissemination of northern mite-resistant stocks.Selection Process

Figure 1 Chewed mites under

l

l

• Promoting Breeding

Inaugural HHBBC at the Purdue University beelaboratory, where 75 queens were in distributed.Breeders from PA, WV, OH, attended the three dayevent.

Grooming behavior is one of the mechanism of defense againstVarroa destructor. Mite grooming behavior is the ability to remove mites and is associated with chewed mites.

Mites are placed on there idiosoma and inspected for damage.

This shows a a response to our selection. We may be near the peak for grooming behavior but need make sure every colony has a high levels as possible by requeening the low chewers. We are working on a better measure for grooming behavior, and have a better correlation between % chewed mites and reduced mite populations in 2014 than we did in 2013, in 2013 the mite populations were higher and more variable between colonies..

Annual queen rearing classes taught at the Purdue bee lab, in 2014 we had 34 students.

Instrumental insemination classes have also been a good resource for breeders to learn more about the importance of selection at the Purdue Bee Lab.

2016 Insemination Fest

PA, OH, WV, MI, IN, IL, KY, TN

130 queens inseminated

COMMUNITY STOCK EVALUATION IN 2014-2015

54 queens, 23 beekeepers,3 commercial sources,

2 Purdue sources.

Commercial hives had 3X more mites

PHOTO: RATNATHAPA

10%

20%

6/27

30%

40%

50%

60%

16/27= 59 %

= 22 %

0%

Commercial Queens IN Mite Biters

Survival 2014 - 2015

10%

20%

30%

40%

50%

60%

7/27= 26 %

17/27= 63 %

*DIED IN WINTER 2014 - 2015

0%

Commercial Queens IN Mite Biters

IN QUEENS MADE 40 LB MORE HONEY

Ten of eleven beekeepers preferred IN mite biters

HAS July 13-15, 2017Evansville, INJuly 13

2016 I.I. Class

Future plans:Cage bee assays for grooming!

Se re Sel Q u e s

0 Redo search when m •p m o"e:d

+.1oux Falls

0

Rochester0 ,

\'\.S 1 City )

I OWADes Moines

0

Cedar Rapid s0

lo w ity

Omaha0

- Jncotn0

Kansas City0

0Lawrertee

Vich it a0

Spr ingf i e ld0

M I S S O U R I

ru lsa0

' ··- - · rOshkoshI

I

Madoson -M-i lwaukee0

·- -Chicago

-0 0N a per v il l e

I

I LLI N OI S - os i ir i ng f ield Champaign

0

St o u is- 0

GrandJ!_apldsRochester

0

IFort r ;ayne

IN D A NAInd ianapol is

0

OH IO0

Columbus

Cmc1l'nat10 W ashington

.o D ILou1sv1lle

0

K E N T U C K YR ic h m o n d

0

...Nashvi l le

0 Map data t 2 0 1 5 Googh!. INEGI

W e lc o m e t o t h e H u n t la b ' s Be e H i v e . W e a r e in v o lv e d i nh o n e y b e e r e s e a r c h a n d e d u c a t i o n .

J uly 9 -11 , 2 0 1 5

W i s y o u w e r e },e r e C 1o l • - 11- rl aiul A p i cu l l u r a l ,:,od e • Y -

-.h e a r t la n d b e e s .o r g

1 4 T H A N N UAL H.A.S. CO N FERENCEf • a h ..r i n g sesa1ons w it h :

Dennis vanEngetsdO<J>, Sue Cob<ty. L a r r y C o nnor , Joe Traynor, C l anm c e Col l isonK i m F lottum, Renata Borba, Roger Hoopln11a ner Ken Schramm , ghan M i lb ra th

c o n f e r e n c es p o n s o r e d

b y :B e e C ul t ur e

b e g to adv. wort.shop,.,

b<JildJng nucs sp l ts ,q-ueen rai.sing .

drone catch tng.new propotis d scover ies

honey tast in gb nn g 3 jars t o swap!0 - - - - -

, . - . . . . . . . ./'lo._ ... . . .. CH ILD REN'S PROGRAMMING :c a nd le - m a k in g, m u r a ls , s e e d b o m b s,,___

P ur d u e U ni v e r s i t I C o l le g e o f A g r icul t u r e I En t o m o lo g y I E x t e n s io n

CoQY.. .dght © 2 0 0 8 , Pu r d u e U n iv e r s i t y , a l l r i g h t s r e s e r v e d . s i t e a u t h o r G r e g H u n tW e b s i te d e v e l o p e d b y t h e E n t o m o l o g y D e p a r t m e n t a t P ur d u e U n iv e r s i t y

I f y o u h a v e t r o u b l e a c c e s s in g t h is p a g e b e c a u s e o f a d is a b ili t y , p l e a s e c o n t a c t t h e W e b m a s t e r .

Events

Do you have a swarm to pick up? Find a local swarm catcher here:http:LLwww.in.govLdnr/entomoloLS755.htm.

When bees swarm the queen and most of the hive beesleave after filling up on honey. Bees in a swarm are usually very gentle but leave them alone! With all those bees flying one could get stuck in your hair.The swann sends out scout bees to find a new cavity to nest in. They usually stay for a few days and thenthey are off to their new home.

Heartland Honey Bee Breeders Coop

National Honey Board Funding

NEW USDA Funding !!!

Jeff Berta - PA Matt Evans – IN Tammy Horn - KYJoe Kovalevski – OH Scott Martin - IL Megan Milbraith - MI Dan O’Hanlon - WV Dave Shenefield - IN Dwight Wells - OH

Greg HuntJennifer Tsuruda

Gladys Andino

Thank you