TEMPERATURE AND CLIMATE IN CHALKBROOD DISEASE José M Flores Departament of Zoology. University of Córdoba. Spain.

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1 TEMPERATURE AND CLIMATE IN CHALKBROOD DISEASE José M Flores Departament of Zoology. University of Córdoba. Spain. ba1flsej@uco.es

2 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

3 CHALKBROOD SYMPTOMS MUMMIES INTO BROOD CELLS, BOTTOM BOARD, HIVE ENTRANCE,

4 CHALKBROOD = A. apis + PREDISPOSING CONDITIONS SOME PREDISPOSING CONDITIONS HAD BEEN RESEARCHED CHILLED BROOD HIGH HUMIDITY ANTIBIOTICS LACK OF POLLEN VERY IMPORTANT PREDISPOSING CONDITION SECONDARY PREDISPOSING CONDITION NON- DEMOSTRATE PREDISPOSING CONDITION IN A SHORT TIME 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: 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: The role of pollen in chalkbrood disease in Apis mellifera: transmission and predisposing conditions (2005). J M Flores, I Gutiérrez, R Espejo. Micologia, 97: Oxytetracicline as predisposing condition for chalkbrood in honey bee (2004). J M Flores, I Gutiérrez, P Puerta. Veterinary Microbiology, 103:

5 WHEN LOW TEMPERATURES TRIGGER CHALKBROOD DEVELOPMENT IN INFECTED BEE BROOD? RESULTS

6 % MUMMIFICATION ± ±16.6 STRESS PERIOD ± ± ±3.4 24h BS 24h AS 48h AS 72h AS CONTROL BROOD STAGE L5 SL Percentage (mean ± sd. Five test)of chalkbrood mummies produced in honey bee larvae after being inoculate with 5x10 5 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: PP

7 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:

8 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:

9 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.

10 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:

11 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:

12 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:

13 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.

14 IS HUMIDITY ONE PREDISPOSING CONDITION FOR CHALKBROOD DISEASE? % MUMMIFICATION ± ± % 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:

15 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.

16 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

17 TEMPERATURE, CLIMATE AND THE BEE COLONIES

18 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.

19 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

20 THANK YOU, VERY MUCH COLOSS (COST), COMITEE, FOR INVITATION AND FOR LISTEN