TEMPERATURE AND CLIMATE IN
CHALKBROOD DISEASE
José M Flores
Departament of Zoology. University of Córdoba.
Spain.
http://www.uco.es/apicultura
[email protected]
CHALKBROOD
A. apis
SPORES
MICELIUM
Chalkbrood in honey bee is a fungal brood
disease caused by Ascosphaera apis.
Spores of A. apis germinate into larval gut
and mycelium invades larval tissues, causing
the died of the brood.
After that, the cadaver becomes dehydrated
like a piece of chalk.
Non sporulated mummies appear white and
the body of sporulated mummies appears
dark-grey
CHALKBROOD SYMPTOMS
MUMMIES INTO BROOD CELLS, BOTTOM BOARD, HIVE ENTRANCE, …
CHALKBROOD = A. apis + PREDISPOSING CONDITIONS
SOME PREDISPOSING CONDITIONS HAD BEEN
RESEARCHED
CHILLED
BROOD
HIGH
HUMIDITY

VERY IMPORTANT
PREDISPOSING
CONDITION
SECONDARY
PREDISPOSING
CONDITION
ANTIBIOTICS
NONDEMOSTRATE
PREDISPOSING
CONDITION IN A
SHORT TIME
LACK OF
POLLEN
MINOR
PREDISPOSING
CONDITION
Chalkbrood development in honeybee brood under controlled conditions (1994). F Puerta, J M Flores, M Bustos, F Padilla y F
Campano. Apidologie 25: 540-545.
Effect of temperature and humidity of sealed brood on chalkbrood development under controller conditions (1996). J M Flores, J A
Ruíz, J M Ruz, F Puerta, M Bustos, F Padilla y F Campano. Apidologie 27: 185- 192.
The role of pollen in chalkbrood disease in Apis mellifera: transmission and predisposing conditions (2005). J M Flores, I Gutiérrez, R
Espejo. Micologia, 97: 1171-1176.
Oxytetracicline as predisposing condition for chalkbrood in honey bee (2004). J M Flores, I Gutiérrez, P Puerta. Veterinary
Microbiology, 103: 195-199.
WHEN LOW TEMPERATURES TRIGGER CHALKBROOD
DEVELOPMENT IN INFECTED BEE BROOD?
RESULTS
% MUMMIFICATION
70
65.5±16.6
59.6±11.8
60
50
40
30
20
10
STRESS
PERIOD
BROOD
STAGE
4.6±3.3
4.9±4.4
4.4±3.4
0
24h BS
L5
24h AS
48h AS
SL
72h AS
CONTROL
PP
Percentage (mean ± sd. Five test)of chalkbrood mummies produced in honey bee larvae
after being inoculate with 5x105 A. apis spores and be chilled in one of 4 periods of
brood development. The control group were not chilling. BS) before capping cells, AS)
after capping cells, L5) fifth larval instar (before capping), SL) spinning larvae and PP)
prepupa.
Chalkbrood development in honeybee brood under controlled conditions (1994). F Puerta, J M Flores, M Bustos, F Padilla y F
Campano. Apidologie 25: 540-545.
WHEN LOW TEMPERATURES TRIGGER CHALKBROOD
DEVELOPMENT IN CONTAMINATED BEE BROOD?
PREVIOUS PREMISE. LARVAE INGESTED A. apis SPORES
ONLY CHILLING BROOD AROUND CAPPING TIME TRIGGER
CHALKBROOD
TO CHILL BROOD BEFORE 8 DAYS, DO NOT CAUSE CHALKBROOD. LARVAE
CAN DIE FOR OTHER CAUSE AND CHALKBROOD APPEARS SECONDARILY.
TO CHILL PREPUPA O LATER DO NOT CAUSE CHALKBROOD. LARVAE CAN
DIE FOR OTHER CAUSE.
Chalkbrood development in honeybee brood under controlled conditions (1994). F Puerta, J M Flores, M Bustos, F Padilla y F
Campano. Apidologie 25: 540-545.
WHATH IS LOW TEMPERATURES FOR CHALKBROOD
DISEASE DEVELOPMENT?
WE WORKED WITH THREE TEMPERATURES AROUND CAPPING
TIME FOR CHALKBROOD RESEARCH: 25ºC, 30ºC AND 35ºC.
Brood chilled before
sealing to 18ºC
(24h)(A) and no
chilled before sealing
(B), and kept after
sealed to 25ºC, 30ºC
and 35ºC.
Effect of temperature and humidity of sealed brood on chalkbrood development under controller conditions (1996). J M Flores, J A
Ruíz, J M Ruz, F Puerta, M Bustos, F Padilla y F Campano. Apidologie 27: 185- 192.
HOW USE EACH TEMPERATURE FOR CHALKBROOD
RESEARCH?
35ºC IS AN EUGENIC TEMPERATURE.
WE KEEP BROOD AFTER SEALING AT 35ºC AS CONTROLS , WE
DO NOT WANT STRESS BY TEMPERATURE OR WE DO NOT WANT
CHALKBROOD DISEASE, ALTHOUGH WE WORK WITH INFECTED
BROOD.
30ºC IS A BORDER TEMPERATURE.
WE KEEP BROOD AFTER SEALING AT 30ºC WHEN WE WANT
KNOW IF ANOTHER FACTOR CAN HELP FOR CHALKBROOD
APPEARS. FOR EXAMPLE TO VALUE HUMIDITY, NUTRITIONAL
STRESS, …
25ºC IS A HIGH STRESS OF TEMPERATURE FOR BEE BROOD.
WE KEEP BROOD AFTER SEALING AT 25ºC WHEN WE WANT
INDUCE CHALKBROOD DISEASE IN INOCULATED BROOD OR WE
WANT KNOW IF BROOD FROM ENYONE COLONY IS INFECTED.
CAN WE SIMPLIFY THE TECNICH FOR CHALKBROOD
DEVELOPMENT?
YES, WE CAN
LATER RESEARCH SHOWED THAT IS NOT NECESSARY CHILL
BEE BROOD BEFORE SEALING FOR CHALKBROOD
DEVELOPMENT.
IT IS ENOUGH TO KEEP BEE BROOD AT 25ºC ALL TIME AFTER
SEALING FOR CHALKBROOD DEVELOPMENT.
ALSO, IT IS ENOUGH TO KEEP BEE BROOD AT 30ºC ALL TIME
AFTER SEALING TO VALUE OTHER FACTORS.
A comparison of methods to experimentally induce chalkbrood disease in honey bee (2004). J M Flores, I Gutiérrez, F Puerta. Spanish
Journal od Agricultural Research, 2: 79-83.
CAN WE USE STRESS AT 25ºC TO ESTUDY OTHER
FACTORS FOR CHALKBROOD DISEASE?
YES, WE CAN
EXAMPLE
WE VALUED IF SPORES OF A. apis INTO FUNDATION CAN
PROVOKE CHALKBROOD.
WE INTRODUCED CONTAMINATES WAX FOUNDATION IN
HEALTHY BEE COLONIES. SOME DAYS AFTER, WE REMOVED
BROOD AFTER CAPPING AND KEPT IT AT 25ºC.
Spores of Ascosphaera apis contained in wax foundation can infect honey bee brood (2005). J M Flores, M Spivak, I Gutiérrez.
Veterinary Microbioly, 108: 141-144.
CAN THE SPORES OF A. apis INTO FOUNDATION COMBS
TRANSMIT CHALKBRD ?
GROUP THREE SHOWED SIGNIFICANTLY MORE CHALKBROOD
THAN GROUP ONE.
SPORES INTO WAX FOUNDATION CAN TRANSMIT CHALKBROOD.
Spores of Ascosphaera apis contained in wax foundation can infect honey bee brood (2005). J M Flores, M Spivak, I Gutiérrez.
Veterinary Microbioly, 108: 141-144.
CAN WE USE STRESS AT 30ºC (temperature border) TO
ESTUDY OTHER FACTORS FOR CHALKBROOD DISEASE?
YES, WE CAN
E. G.
WE VALUE THE HUMIDITY AS PREDISPOSING CONDITION.
WE KEEP INFECTED BEE BROOD AT 30ºC (24 H BEFORE
SEALING AND THE PERIOD OF 6 DAYS AFTER SEALING) WITH
HIGH (87% RH) AND LOW (68% RH) HUMIDITY.
IS HUMIDITY ONE PREDISPOSING CONDITION FOR
CHALKBROOD DISEASE?
% MUMMIFICATION
7.75±6.57
8
7
6
5
4
3
2
0.95±1.70
1
0
68% RH
87% RH
HUMIDITY CAN BE A SECUNDARY PREDISPOSING CONDITION.
TO KEEP BEE BROOD AT 30ºC HELPED TO FOUND DIFFERENTS
PERCETAGES OF MUMMIFICATION BETWEEN THE TWO
HUMIDITIES.
Effect of temperature and humidity of sealed brood on chalkbrood development under controller conditions (1996). J M Flores, J A
Ruíz, J M Ruz, F Puerta, M Bustos, F Padilla y F Campano. Apidologie 27: 185- 192.
CAN WE KEEP BEE BROOD AT 35ºC TO ESTUDY OTHER
FACTORS FOR CHALKBROOD DISEASE?
YES, WE CAN
TEMPERATURE 35ºC CAN BE USED AS CONTROL.
WE WOULD KEEP BEE BROOD AT 35ºC TO ASSURE THAT
TEMPERATURE IS NOT AN STRESS DURING TEST WHEN WE WANT
VALUE OTHER MAIN PREDISPOSING CONDITIONS.
WHAT HAPPENS IF WE KEEP BEE BROOD AT
TEMTERATURES LOWER TO 25ºC?
TEMPERATURES LOWER 25ºC (FOR EXAMPLE 20/22ºC), BEE
BROOD DOES NOT DEVELOP
AND CHALKBROOD NEITHER .
BEE BROOD DIE AND CHALKBROOD CAN APPEARS AS
SECUNDARY EFFECT.
Non publiched results
TEMPERATURE, CLIMATE AND
THE BEE COLONIES
WHY, HOW AND WHEN TEMPERATURE CAN INDUCE
CHALKBROOD DISEASE?
 LOW TEMPERATURES AROUND CAPPING TIME IS ENOUGH TO
PROVOKE CHALKBROOD.
CHILLING BROOD INTO BEE HIVE CAN BE BY:
Fall of enviromental temperatures.
High increase of brood versus number of worker bees.
Reduction of worker bees due to other diseases o toxics.
Bad handling of honey bees colonies.
CCD provoke fast redution of worker honey bees.
 CLIMATIC CHANGE CAN INDUCE CHALKBROOD BY:
Alternative short periods (few days) of hot and cold temperatures.
High temperatures in autumn or winter followed by cold wether.
AND DAMP WETHER?
 HUMIDITY IS A SECUNDARY PREDISPOSING CONDITION. WE
ARE NO SURE OF THE ROLE OF THE DAMP WETHER.
HUMIDITY INTO BE HIVE CAN BE:
Entry water to the bee hive.
A big flowering.
IT IS NECESSARY STUDY DAMP WETHER IN
CHALKBROOD DEVELOPMENT
THANK YOU, VERY MUCH
COLOSS (COST), COMITEE, FOR INVITATION AND FOR
LISTEN
http://www.uco.es/apicultura
[email protected]