Honeybee - RIRDC Completed Projects in 2001--2002 & Research in Progress as at June 2002

Project Title	Improving queen bee production
RIRDC Project No.:	CSE-85A
Start Date:	01/07/99
Finish Date:	31/07/02
Researcher:	Dr. Denis Anderson
Organisation:	CSIRO Entomology GPO Box 1700 CANBERRA ACT 2601
Phone:	(02) 6246 4148
Fax:	(02) 6246 4000
Email:	Denis.Anderson@csiro.au
Objectives	To improve the profitability of commercial queen producers by improving queen performance. This will be achieved by: 1. Determining whether nutritional supplements significantly improve queen and drone quality; 2. Determining the factors which are responsible for causing disappearing disorder.
Current Progress	*Effects of nutrient supplements on queen banks* In the autumn of 2001 tests were carried out to determine whether nutrient supplements fed to queen banks affected the quality of the queens banked in them. Two hundred sister-queens in small cages were banked in groups of 20 in one of 10 standardized queen banks. Groups of 2 banks were then fed one of the following supplements on a continuous basis for 2 months: (a) no supplement (control), (b) soya-flour based supplement, (c) commercially available pollen patty, (d) vitamins in 60% sucrose solution and (e) 60% sucrose solution. After 2 months the banked queens were tested for: (1) weight, (2) number of ovarioles, (3) diameter of spermathecae, (4) number of spermatozoa in spermathecae, and (5) nosema levels. There was no significant differences in the quality of the queens kept in the banks that were fed the different supplements. Tests in the spring of 2002 will focus of the effects of feeding nutrient supplements to drone-rearing colonies. *Disappearing Disorder* During the 2000/01 season, brood, pollen and nectar samples were collected from colonies affected with disappearing disorder at a single apiary in Queensland. Tests on these samples showed some evidence that elevated levels of iron may have contributed to the cause of the disorder. Results from tests conducted on colonies affected with the disorder at Gladstone during the 2001/02 season are still being finalised.

RESEARCH IN PROGRESS REPORT 2002

Project Title Introduction and early performance of queen bees - some factors affecting success

RIRDC Project No.: DAN-182A

Start Date: 01/09/99

Finish Date: 31/07/02

Researcher: Mr. John Rhodes

Organisation: NSW Department of Agriculture
RMB 944
Calala Lane
TAMWORTH NSW 2340

Phone: (02) 6763 1206

Fax: (02) 6763 1222

Email: john.rhodes@agric.nsw.gov.au

Objectives 1. To determine whether the age of the queen bee at introduction influences introduction success.
2. To identify factors affecting sperm counts of sister queen bees mated under the same mating conditions and to investigate relationships between low spem counts and queen failure following introduction.
3. To examine the effects of temperature and humidity during transport of queen bees on their introduction success.
4. A comparison of sperm counts between sister bees mated in a commercial queen bee breeder's apiary and between those mated in a commercial honey producer's queen bee breeder's apiary.
5. Development of a method for preserving adult drones which would allow accurate sperm counts to be carried out on those drones.
6. To determine differences in (i) introduction and short term survival rates, and (ii) the number of sperm in the spermatheca, between queen bees held in (i) mating nuclei, and (ii) queen banks, from 17 days of age until 24 and 31 days of age before being introduced into commercial honey hives.
7. To examine the effects of temperature and humidity during transport of queen bees on their introduction success.

Current Progress Points 1, 2, 3, 4, 6 and 7 have been completed, Point 5 was not able to be achieved successfully and an alternative method has been found. The final report for DAN 182A is being prepared.
Point 1: Increased age of the queen at introduction improved introduction success, 21-28 days old was the most suitable age for introduction. One mandibular gland pheromone showed a positive correlation with both queen age and introduction success.
Point 2: Based on physical measurements of ovariole numbers, spermatheca diameter and queen weight, all queens used in the project were of a high standard when compared with USA and NZ data. Sperm counts for queens caught at all ages, but particularly at 7 and 14 days of age, were generally below the acceptable levels of 3 million sperm for survival >12 months, and 4.5 million sperm considered suitable for a commercially reared queen based on USA data.
Point 3, Point 7: No relationship found, to date, between temperature and humidity data queens were subjected to during transport and queen survival and performance.
Point 4:No significant difference was found between sister queens mated in a queen breeder`s or a honey producer`s mating apiary.
Point 5: A satisfactory method for preserving adult drones to allow examination at a later date was not achieved. A process where freshly caught drones can be examined to provide the required data has been determined.
Point 6: The queen banking project has been completed and data is being analysed. Compared to data from the 2 previous years, sperm counts were higher but still averaged below 4.5 million/queen. Queen survival rates were also higher with queens caught at 24 and 31 days old higher than 17 day old. Banking reduced survival for 24 d.o. queens but not for 31 d.o. queens.

RESEARCH IN PROGRESS REPORT 2002

Project Title Device for finding queen bees in managed beehives

RIRDC Project No.: PFP-1A

Start Date: 01/02/01

Finish Date: 28/02/03

Researcher: Mr. Alan Roberts

Organisation: Podargus Farm Products
Green Mountain Electronics
Boundary Creek Road
BENTLEY NSW 2480

Phone: (02) 6663 5224

Fax: --

Email: alanrbts@fan.net.au

Objectives To design and prove an electronic device capable of pinpointing the position of queen bees in managed beehives.

Current Progress The aim of the project is to attach a small device on a honey bee queen that will allow a scanning tool to pinpoint her position in the hive. This would allow faster finding of those queens (which have not been superseded) during requeening. It will also enable a quick check to be undertaken (from outside the hive) to ascertain whether the original queen is still in attendance. A benefit for both commercial beekeepers and researchers.
The project has not advanced as quickly as anticipated for two unexpected reasons. The principal researcher has a serious illness which has slowed progress considerably.
There has also been reluctance, by some electronic component manufacturers, to supply their products in the small quantities required for research. All components for a preliminary test have now been obtained.
Both researchers attended the "Austronics 2001" trade show and workshops to view latest components and electronic assembly techniques.
It is envisioned that assembly and preliminary testing will take place by the end of May, and then wider testing during the spring of 2002 under a range of hive conditions.

RESEARCH IN PROGRESS REPORT 2002

Project Title A study of Gluconobacter - gluconic acid producing bacteria, symbionts of bees: development of biological control for chalkbrood

RIRDC Project No.: ANU-58A

Start Date: 01/01/02

Finish Date: 30/04/04

Researcher: Dr. Murali Nayudu

Organisation: Australian National University
School of Botany and Zoology
Faculty of Science
CANBERRA ACT 0200

Phone: (02) 6125 3643

Fax: (02) 6125 5573

Email: Murali.Nayudu@anu.edu.au

Objectives To isolate and characterise from different Australian bee hives the bacteria that produce gluconic acid (identified by us as an anti-fungal agent). The results of this strategic basic research will provide specific information to carry out applied research in the future to develop biological control of chalkbrood disease.

Current Progress Bacterial species such as Bacillus have been frequently isolated from European honeybee (Apis mellifera) collected from a variety of regions in other parts of the world. The nature of this bee-bacterial relationship is poorly understood. Hydrogen peroxide has been identified as a strong anti-microbial agent. More recently gluconic acid has been shown to be a powerful anti-fungal agent. The relative contribution of these two different sources to the hydrogen peroxide and gluconic acid present in honey is unknown.
The aim of this project is to study the nature of Australian gluconic acid production of bacteria that associate with honeybees. At the start of this project we have shown by isolation that a diverse range of bacterial species associate with Australian honey bees. Many of these are acid producing strains, and several may be producing the anti-fungal agent gluconic acid. Over 50% of the bacterial strains isolated from honeybees inhibit chalkbrood. This suggests that there is selection for bacteria that can compete in the intestinal tract of honeybees with fungi like chalkbrood.
In conclusion the results so far strongly suggest there is a potential to utilise these Australian bee bacteria to obtain a biological control for chalk brood.

RESEARCH IN PROGRESS REPORT 2002

Project Title Clarification of aspects of Varroa reproduction - first stage of a possible new control method

RIRDC Project No.: CSE-87A

Start Date: 01/07/01

Finish Date: 30/08/03

Researcher: Dr. Denis Anderson

Organisation: CSIRO Entomology
GPO Box 1700
CANBERRA ACT 2601

Phone: (02) 6246 4148

Fax: (02) 6246 4000

Email: Denis.Anderson@csiro.au

Objectives To determine the hormone profiles in the blood of pre and post-pupal stages of Apis mellifera and Apis cerana drone brood in Java, Indonesia, and in the blood of Varroa mites infesting those stages. This information will form a basis for developing a possible new method for controlling Varroa destructor on Apis mellifera.

Current Progress This project aims to gain a better understanding of why the Java genotype of Varroa jacobsoni can reproduce on drone brood of its native Apis cerana in Java but not on the drone brood of Apis mellifera. Understanding the underlying mechanisms could lead to novel ways of controlling those Varroa genotypes that can reproduce on A. mellifera.
Clearances and permits have been obtained for moving bee and mite material from Indonesia to Australia. Colonies of A. cerana and A. mellifera have been established at Paung Panjang and Bogor and these will be used to source all bee and mite material throughout the course of the project.
In the initial stages, tests are being conducted to determine the best method for extracting hormones from the samples collected in Java. Hence, blood samples were collected from prepupal and early pupal stages of A. cerana and A. mellifera in Java from 1-5 October and again from the 1-4 April 2002. The October samples were placed in liquid nitrogen prior to being shipped, while the April samples were placed in methanol. Dr M. Lacey at CSIRO Entomology is currently extracting hormones from these samples.

RESEARCH IN PROGRESS REPORT 2002

Project Title The sensitivity of Australian honey bee bacterial pathogens to fatty acids

RIRDC Project No.: DAN-193A

Start Date: 01/08/01

Finish Date: 31/07/02

Researcher: Dr. Michael Hornitzky

Organisation: NSW Department of Agriculture
Elizabeth Macarthur Agricultural Institute
Private Mail Bag 8
CAMDEN NSW 2570

Phone: (02) 4640 6311

Fax: (02) 4640 6400

Email: michael.hornitzky@agric.nsw.gov.au

Objectives To determine the sensitivity of the two major bacterial honey bee pathogens in Australia, ie. Melissococcus pluton (the cause of European foulbrood [EFB]) and Paenibaccillus larvae subsp. larvae (the cause of American foulbrood [AFB]) to a range of fatty acids including linoleic acid.

Current Progress The activity of 28 fatty acids was tested against 13 Paenibacillus larvae subsp. larvae isolates from NSW, South Australia, Tasmania, Western Australia, Queensland, Northern Territory, New Zealand and China. These fatty acids were also tested against 10 Melissococcus pluton isolates from NSW, South Australia, Tasmania, Queensland and the United Kingdom. Fifteen fatty acids had activity against P. l. larvae in the following decreasing order of activity; undecanoic, homo-g -linolenic, capric, 7, 10, 13, 16-docosatetraenoic, 13, 16, 19-docosatrienoic, linolenic, linoleic, myristoleic, lauric, 11, 14-eicosadienoic, ricinoleic, 13, 16-docosadienoic, ricinelaidic, arachidonic and g -linolenic acid. Eight fatty acids showed activity against M. pluton in the following decreasing order of activity; undecanoic, lauric, myristoleic, 13, 16, 19-docosatrienoic, homo-g -linolenic, ricinoleic, ricinelaidic and myristic. There was generally little variation in the sensitivity of the various strains of either P. l. larvae or M. pluton to any specific fatty acid.
The activity of the fatty acids against P. l. larvae was substantially less than previously reported.

RESEARCH IN PROGRESS REPORT 2002

Project Title European Foulbrood - investigating control measures

RIRDC Project No.: DAV-157A

Start Date: 07/12/98

Finish Date: 30/09/02

Researcher: Mr. Russell Goodman

Organisation: Department of Natural Resources & Environment (Vic)
Institute for Horticultural Development
Private Bag 15, SCORESBY BUSINESS CENTRE VIC 3176

Phone: (03) 9210 9222

Fax: (03) 9800 3521

Email: russell.goodman@nre.vic.gov.au

Objectives a) To protect the apiary industry's continued access to domestic and export honey markets by reducing or eliminating industry's dependence on oxtetracycline hydrochloride (OTC) for the control of the bacterial honeybee brood disease, European Foulbrood (EFB) (Melissococcus pluton).
b) To determine the efficacy of reduced doses of OTC and use of OTC extender patties for the control of EFB and to determine if these measures reduce or eliminate the occurrence of OTC residues in honey.
c) To identify and develop alternative, non-antibiotic measures for control of EFB by investigating, primarily, the effect of enhanced honeybee colony nutrition and changed pH of honeybee larval guts.
d) To obtain a greater understanding of active and latent infections of M. pluton and Paenibacillus alvei (a common secondary invader) in honeybee larvae and to develop new Polymerase Chain Reaction (PCR) methodologies for detection of M. pluton as a necessary prerequisite and support of the preceding aim (c)

Current Progress Field trials
OTC (1.0g, 0.5g and 0.3g) was applied in either caster sugar or water to honey bee colonies to determine the concentration of residues in honey that was later extracted from the hives.
The incidence of EFB in 3 groups of colonies fed high protein pollen cakes, treated with 1g OTC or untreated controls was determined. The number of diseased colonies and the severity of disease was lower in colonies fed protein when compared to the controls.
Laboratory investigations
Honey extracted from hives treated with 1g OTC was decanted into vials and placed in 25ºC, 30ºC, 35ºC and 35ºC incubators. Vials were removed from each incubator fortnightly and analysed for OTC residues.
The minimum infectious dose for EFB was determined by inoculating laboratory reared larvae with M. pluton. These methodologies provide a rapid assay for determining the efficacy of potential treatments for control of EFB and other diseases affecting honey bee larvae.
Preliminary studies on the potential for larval midgut pH to influence susceptibility to EFB were commenced. Examination of histology sections of EFB infected larvae as part of studies on the pathogenesis of M. pluton confirmed the organism not only multiplies in the larval gut but also invades the peritrophic membrane.

RESEARCH IN PROGRESS REPORT 2002

Project Title	Predicting the productivity of honeybees from the nutritional value of pollen
RIRDC Project No.:	ANU-57A
Start Date:	01/12/01
Finish Date:	31/12/04
Researcher:	Mr. Ian Wallis
Organisation:	Australian National University Division of Botany & Zoology CANBERRA ACT 0200
Phone:	(02) 6249 2533
Fax:	(02) 6249 5573
Email:	ian.wallis@anu.edu.au
Objectives	To devise a rapid method for explaining the nutritional status and productivity of a colony of bees from the nutritional value of pollen they eat.
Current Progress	The first objective of this project, started on 1/12/2001, was to appoint a PhD student. This took longer than anticipated. We started searching in July 2001 and Susan Burrows, a graduate of the University of Sydney, started in February 2002. Our second objective was to test whether near infrared reflectance spectroscopy (NIRS) could predict the nitrogen (crude protein) and amino acid content of pollen. Briefly, the procedure involves scanning samples with NIRS, analysing the samples with traditional chemistry and then statistically relating the two data sets. The resulting prediction equation enables future samples to be analysed quickly and cheaply using NIRS alone. We studied pollen (72 samples of 50 species collected between August 1995 and December 1998) donated by Doug Somerville (NSW Dept of Agriculture). While we could justify repeating the nitrogen analyses, we could not justify the expense of reanalysing them for amino acids. Thus, we did NIRS scans of 72 samples, which we related to our nitrogen analyses but to Doug Somerville's published values for amino acids. This pilot study produced an excellent model for predicting nitrogen but poorer models for amino acids. This implies some degradation of amino acids during storage. We are now preparing a website and purchasing bee equipment.

RESEARCH IN PROGRESS REPORT 2002

Project Title Production of a publication on honeybee nutrition in Australia - 'Fat bees/skinny bees'

RIRDC Project No.: DAN-186A

Start Date: 01/01/00

Finish Date: 31/10/02

Researcher: Mr. Doug Somerville

Organisation: NSW Department of Agriculture
PO Box 389
GOULBURN NSW 2580

Phone: (02) 4828 6619

Fax: (02) 4822 3261

Email: doug.somerville@agric.nsw.gov.au

Objectives To produce an extension publication on honey bee nutrition, incorporating research findings from past RIRDC projects, literature searches and anecdotal examples of applications in the Australian context in a format that will be readily understood and adopted by beekeepers.

Current Progress A paper on the Mineral Content of Bee Collected Pollen has been accepted by the Australian Journal of Experimental Agriculture. Information on all areas of Honeybee nutrition has been gathered and is in the process of being compiled into sections on the major nutritional components impacting on the diet of Honeybees.
Further case studies, particularly for Tasmania, WA, Queensland and Victoria are scheduled to be collected in the winter and spring of 2002.
The publication will be delayed if necessary, within reason to include the results of a number of current RIRDC funded projects, including data on Fatty Acid Analysis of Pollen’s, the effects of Fatty Acids on Honeybee pathogens, the use of protein supplements in the role of Queen Bee production system’s and the implications of minerals in pollen and nectar, linked to a condition referred to as Disappearing Disorder.

RESEARCH IN PROGRESS REPORT 2002

Project Title An Australian survey of pollens for their fatty acid composition

RIRDC Project No.: DAW-100A

Start Date: 01/05/01

Finish Date: 01/12/04

Researcher: Mr. Robert Manning

Organisation: Department of Agriculture (WA)
Locked Bag No 4
BENTLEY DELIVERY CENTRE WA 6983

Phone: (08) 9368 3567

Fax: (08) 9474 1295

Email: rmanning@agric.wa.gov.au

Objectives · Identify fatty acids of pollens from twenty major honey and pollen producing plant species from each State of Australia.
· Categorise the importance of each species and compare fatty acid profiles from previous studies.
· Determine the effects of long term cold storage on the fatty acids of pollens.

Current Progress Approximately 130 pollen samples have been analysed from 40 major honey and pollen producing species. Samples have come from Western Australia, Queensland, NSW and Tasmania. With the drought and honey shortage being experienced nationally the pollen collections have been poor when compared to project expectations. However, as a speaker at 3 Beekeeper annual general meetings (NSW, VIC & SA) over the next couple of months, the pollen collections should improve once beekeepers further understand the project.
The cold storage tests have also commenced.

RESEARCH IN PROGRESS REPORT 2002

Project Title Natural resource database for the South Australian apiary industry

RIRDC Project No.: DEH-1A

Start Date: 05/07/99

Finish Date: 31/05/02

Researcher: Dr. Chris Holden

Organisation: Department of Environment, Heritage and Aboriginal Affairs (SA)
Biodiversity Conservation Program
Heritage and Biodiversity Division
PO Box 1047
ADELAIDE SA 5000

Phone: (08) 8204 8736

Fax: (08) 8204 8889

Email: holden.chris@saugov.sa.gov.au

Objectives To create a database of floral resource information for the South Australian apiary industry, which includes details of: 1. the floral resource base on which the industry is dependent, including the reliance of the industry on native vegetation and the most valuable and reliable floral species for honeybees;
2. the distribution of various native plant species as they relate to beekeeping;
3. the frequency with which these resources are used, the land tenure on which they currently exist, and the relative values for honey and pollen as they relate to honeybee nutritional requirements and honey production; and
4. estimates of the value of currently used apiary sites on both private and Crown lands in terms of dollar value or as a percentage of the State annual production.

Current Progress The researcher Dr David Paton has employed research assistant Emma Crossfield to compile the report. Subsequent to the poor response to the initial questionnaire (sent to 237 registered beekeepers), information was distributed through the South Australian Apiarists Association newsletter, meetings arranged with small collections of beekeepers, a simpler form distributed to non-respondents, and finally phone-calls to those beekeepers who had still not responded. Of the beekeepers approached, 50% have responded. This accounts for 30% of the hives in South Australia and suggests a higher proportion of smaller operators had responded. At best, another 20% of beekeepers may provide information. At least 10% and perhaps as high as 30% of South Australian beekeepers refuse to provide information.
Map production will be finalised in May 2002.
This project is on track for publication in June 2002.

Project Title	Introduction and early performance of queen bees - some factors affecting success
RIRDC Project No.:	DAN-182A
Start Date:	01/09/99
Finish Date:	31/07/02
Researcher:	Mr. John Rhodes
Organisation:	NSW Department of Agriculture RMB 944 Calala Lane TAMWORTH NSW 2340
Phone:	(02) 6763 1206
Fax:	(02) 6763 1222
Email:	john.rhodes@agric.nsw.gov.au
Objectives	1. To determine whether the age of the queen bee at introduction influences introduction success. 2. To identify factors affecting sperm counts of sister queen bees mated under the same mating conditions and to investigate relationships between low spem counts and queen failure following introduction. 3. To examine the effects of temperature and humidity during transport of queen bees on their introduction success. 4. A comparison of sperm counts between sister bees mated in a commercial queen bee breeder's apiary and between those mated in a commercial honey producer's queen bee breeder's apiary. 5. Development of a method for preserving adult drones which would allow accurate sperm counts to be carried out on those drones. 6. To determine differences in (i) introduction and short term survival rates, and (ii) the number of sperm in the spermatheca, between queen bees held in (i) mating nuclei, and (ii) queen banks, from 17 days of age until 24 and 31 days of age before being introduced into commercial honey hives. 7. To examine the effects of temperature and humidity during transport of queen bees on their introduction success.
Current Progress	Points 1, 2, 3, 4, 6 and 7 have been completed, Point 5 was not able to be achieved successfully and an alternative method has been found. The final report for DAN 182A is being prepared. Point 1: Increased age of the queen at introduction improved introduction success, 21-28 days old was the most suitable age for introduction. One mandibular gland pheromone showed a positive correlation with both queen age and introduction success. Point 2: Based on physical measurements of ovariole numbers, spermatheca diameter and queen weight, all queens used in the project were of a high standard when compared with USA and NZ data. Sperm counts for queens caught at all ages, but particularly at 7 and 14 days of age, were generally below the acceptable levels of 3 million sperm for survival >12 months, and 4.5 million sperm considered suitable for a commercially reared queen based on USA data. Point 3, Point 7: No relationship found, to date, between temperature and humidity data queens were subjected to during transport and queen survival and performance. Point 4:No significant difference was found between sister queens mated in a queen breeder`s or a honey producer`s mating apiary. Point 5: A satisfactory method for preserving adult drones to allow examination at a later date was not achieved. A process where freshly caught drones can be examined to provide the required data has been determined. Point 6: The queen banking project has been completed and data is being analysed. Compared to data from the 2 previous years, sperm counts were higher but still averaged below 4.5 million/queen. Queen survival rates were also higher with queens caught at 24 and 31 days old higher than 17 day old. Banking reduced survival for 24 d.o. queens but not for 31 d.o. queens.

Project Title	Device for finding queen bees in managed beehives
RIRDC Project No.:	PFP-1A
Start Date:	01/02/01
Finish Date:	28/02/03
Researcher:	Mr. Alan Roberts
Organisation:	Podargus Farm Products Green Mountain Electronics Boundary Creek Road BENTLEY NSW 2480
Phone:	(02) 6663 5224
Fax:	--
Email:	alanrbts@fan.net.au
Objectives	To design and prove an electronic device capable of pinpointing the position of queen bees in managed beehives.
Current Progress	The aim of the project is to attach a small device on a honey bee queen that will allow a scanning tool to pinpoint her position in the hive. This would allow faster finding of those queens (which have not been superseded) during requeening. It will also enable a quick check to be undertaken (from outside the hive) to ascertain whether the original queen is still in attendance. A benefit for both commercial beekeepers and researchers. The project has not advanced as quickly as anticipated for two unexpected reasons. The principal researcher has a serious illness which has slowed progress considerably. There has also been reluctance, by some electronic component manufacturers, to supply their products in the small quantities required for research. All components for a preliminary test have now been obtained. Both researchers attended the "Austronics 2001" trade show and workshops to view latest components and electronic assembly techniques. It is envisioned that assembly and preliminary testing will take place by the end of May, and then wider testing during the spring of 2002 under a range of hive conditions.

Project Title	A study of Gluconobacter - gluconic acid producing bacteria, symbionts of bees: development of biological control for chalkbrood
RIRDC Project No.:	ANU-58A
Start Date:	01/01/02
Finish Date:	30/04/04
Researcher:	Dr. Murali Nayudu
Organisation:	Australian National University School of Botany and Zoology Faculty of Science CANBERRA ACT 0200
Phone:	(02) 6125 3643
Fax:	(02) 6125 5573
Email:	Murali.Nayudu@anu.edu.au
Objectives	To isolate and characterise from different Australian bee hives the bacteria that produce gluconic acid (identified by us as an anti-fungal agent). The results of this strategic basic research will provide specific information to carry out applied research in the future to develop biological control of chalkbrood disease.
Current Progress	Bacterial species such as Bacillus have been frequently isolated from European honeybee (Apis mellifera) collected from a variety of regions in other parts of the world. The nature of this bee-bacterial relationship is poorly understood. Hydrogen peroxide has been identified as a strong anti-microbial agent. More recently gluconic acid has been shown to be a powerful anti-fungal agent. The relative contribution of these two different sources to the hydrogen peroxide and gluconic acid present in honey is unknown. The aim of this project is to study the nature of Australian gluconic acid production of bacteria that associate with honeybees. At the start of this project we have shown by isolation that a diverse range of bacterial species associate with Australian honey bees. Many of these are acid producing strains, and several may be producing the anti-fungal agent gluconic acid. Over 50% of the bacterial strains isolated from honeybees inhibit chalkbrood. This suggests that there is selection for bacteria that can compete in the intestinal tract of honeybees with fungi like chalkbrood. In conclusion the results so far strongly suggest there is a potential to utilise these Australian bee bacteria to obtain a biological control for chalk brood.

Project Title	Clarification of aspects of Varroa reproduction - first stage of a possible new control method
RIRDC Project No.:	CSE-87A
Start Date:	01/07/01
Finish Date:	30/08/03
Researcher:	Dr. Denis Anderson
Organisation:	CSIRO Entomology GPO Box 1700 CANBERRA ACT 2601
Phone:	(02) 6246 4148
Fax:	(02) 6246 4000
Email:	Denis.Anderson@csiro.au
Objectives	To determine the hormone profiles in the blood of pre and post-pupal stages of Apis mellifera and Apis cerana drone brood in Java, Indonesia, and in the blood of Varroa mites infesting those stages. This information will form a basis for developing a possible new method for controlling Varroa destructor on Apis mellifera.
Current Progress	This project aims to gain a better understanding of why the Java genotype of Varroa jacobsoni can reproduce on drone brood of its native Apis cerana in Java but not on the drone brood of Apis mellifera. Understanding the underlying mechanisms could lead to novel ways of controlling those Varroa genotypes that can reproduce on A. mellifera. Clearances and permits have been obtained for moving bee and mite material from Indonesia to Australia. Colonies of A. cerana and A. mellifera have been established at Paung Panjang and Bogor and these will be used to source all bee and mite material throughout the course of the project. In the initial stages, tests are being conducted to determine the best method for extracting hormones from the samples collected in Java. Hence, blood samples were collected from prepupal and early pupal stages of A. cerana and A. mellifera in Java from 1-5 October and again from the 1-4 April 2002. The October samples were placed in liquid nitrogen prior to being shipped, while the April samples were placed in methanol. Dr M. Lacey at CSIRO Entomology is currently extracting hormones from these samples.

Project Title	The sensitivity of Australian honey bee bacterial pathogens to fatty acids
RIRDC Project No.:	DAN-193A
Start Date:	01/08/01
Finish Date:	31/07/02
Researcher:	Dr. Michael Hornitzky
Organisation:	NSW Department of Agriculture Elizabeth Macarthur Agricultural Institute Private Mail Bag 8 CAMDEN NSW 2570
Phone:	(02) 4640 6311
Fax:	(02) 4640 6400
Email:	michael.hornitzky@agric.nsw.gov.au
Objectives	To determine the sensitivity of the two major bacterial honey bee pathogens in Australia, ie. Melissococcus pluton (the cause of European foulbrood [EFB]) and Paenibaccillus larvae subsp. larvae (the cause of American foulbrood [AFB]) to a range of fatty acids including linoleic acid.
Current Progress	The activity of 28 fatty acids was tested against 13 Paenibacillus larvae subsp. larvae isolates from NSW, South Australia, Tasmania, Western Australia, Queensland, Northern Territory, New Zealand and China. These fatty acids were also tested against 10 Melissococcus pluton isolates from NSW, South Australia, Tasmania, Queensland and the United Kingdom. Fifteen fatty acids had activity against P. l. larvae in the following decreasing order of activity; undecanoic, homo-g -linolenic, capric, 7, 10, 13, 16-docosatetraenoic, 13, 16, 19-docosatrienoic, linolenic, linoleic, myristoleic, lauric, 11, 14-eicosadienoic, ricinoleic, 13, 16-docosadienoic, ricinelaidic, arachidonic and g -linolenic acid. Eight fatty acids showed activity against M. pluton in the following decreasing order of activity; undecanoic, lauric, myristoleic, 13, 16, 19-docosatrienoic, homo-g -linolenic, ricinoleic, ricinelaidic and myristic. There was generally little variation in the sensitivity of the various strains of either P. l. larvae or M. pluton to any specific fatty acid. The activity of the fatty acids against P. l. larvae was substantially less than previously reported.

Project Title	European Foulbrood - investigating control measures
RIRDC Project No.:	DAV-157A
Start Date:	07/12/98
Finish Date:	30/09/02
Researcher:	Mr. Russell Goodman
Organisation:	Department of Natural Resources & Environment (Vic) Institute for Horticultural Development Private Bag 15, SCORESBY BUSINESS CENTRE VIC 3176
Phone:	(03) 9210 9222
Fax:	(03) 9800 3521
Email:	russell.goodman@nre.vic.gov.au
Objectives	a) To protect the apiary industry's continued access to domestic and export honey markets by reducing or eliminating industry's dependence on oxtetracycline hydrochloride (OTC) for the control of the bacterial honeybee brood disease, European Foulbrood (EFB) (Melissococcus pluton). b) To determine the efficacy of reduced doses of OTC and use of OTC extender patties for the control of EFB and to determine if these measures reduce or eliminate the occurrence of OTC residues in honey. c) To identify and develop alternative, non-antibiotic measures for control of EFB by investigating, primarily, the effect of enhanced honeybee colony nutrition and changed pH of honeybee larval guts. d) To obtain a greater understanding of active and latent infections of M. pluton and Paenibacillus alvei (a common secondary invader) in honeybee larvae and to develop new Polymerase Chain Reaction (PCR) methodologies for detection of M. pluton as a necessary prerequisite and support of the preceding aim (c)
Current Progress	Field trials OTC (1.0g, 0.5g and 0.3g) was applied in either caster sugar or water to honey bee colonies to determine the concentration of residues in honey that was later extracted from the hives. The incidence of EFB in 3 groups of colonies fed high protein pollen cakes, treated with 1g OTC or untreated controls was determined. The number of diseased colonies and the severity of disease was lower in colonies fed protein when compared to the controls. Laboratory investigations Honey extracted from hives treated with 1g OTC was decanted into vials and placed in 25ºC, 30ºC, 35ºC and 35ºC incubators. Vials were removed from each incubator fortnightly and analysed for OTC residues. The minimum infectious dose for EFB was determined by inoculating laboratory reared larvae with M. pluton. These methodologies provide a rapid assay for determining the efficacy of potential treatments for control of EFB and other diseases affecting honey bee larvae. Preliminary studies on the potential for larval midgut pH to influence susceptibility to EFB were commenced. Examination of histology sections of EFB infected larvae as part of studies on the pathogenesis of M. pluton confirmed the organism not only multiplies in the larval gut but also invades the peritrophic membrane.

Project Title	Production of a publication on honeybee nutrition in Australia - 'Fat bees/skinny bees'
RIRDC Project No.:	DAN-186A
Start Date:	01/01/00
Finish Date:	31/10/02
Researcher:	Mr. Doug Somerville
Organisation:	NSW Department of Agriculture PO Box 389 GOULBURN NSW 2580
Phone:	(02) 4828 6619
Fax:	(02) 4822 3261
Email:	doug.somerville@agric.nsw.gov.au
Objectives	To produce an extension publication on honey bee nutrition, incorporating research findings from past RIRDC projects, literature searches and anecdotal examples of applications in the Australian context in a format that will be readily understood and adopted by beekeepers.
Current Progress	A paper on the Mineral Content of Bee Collected Pollen has been accepted by the Australian Journal of Experimental Agriculture. Information on all areas of Honeybee nutrition has been gathered and is in the process of being compiled into sections on the major nutritional components impacting on the diet of Honeybees. Further case studies, particularly for Tasmania, WA, Queensland and Victoria are scheduled to be collected in the winter and spring of 2002. The publication will be delayed if necessary, within reason to include the results of a number of current RIRDC funded projects, including data on Fatty Acid Analysis of Pollen’s, the effects of Fatty Acids on Honeybee pathogens, the use of protein supplements in the role of Queen Bee production system’s and the implications of minerals in pollen and nectar, linked to a condition referred to as Disappearing Disorder.

Project Title	An Australian survey of pollens for their fatty acid composition
RIRDC Project No.:	DAW-100A
Start Date:	01/05/01
Finish Date:	01/12/04
Researcher:	Mr. Robert Manning
Organisation:	Department of Agriculture (WA) Locked Bag No 4 BENTLEY DELIVERY CENTRE WA 6983
Phone:	(08) 9368 3567
Fax:	(08) 9474 1295
Email:	rmanning@agric.wa.gov.au
Objectives	· Identify fatty acids of pollens from twenty major honey and pollen producing plant species from each State of Australia. · Categorise the importance of each species and compare fatty acid profiles from previous studies. · Determine the effects of long term cold storage on the fatty acids of pollens.
Current Progress	Approximately 130 pollen samples have been analysed from 40 major honey and pollen producing species. Samples have come from Western Australia, Queensland, NSW and Tasmania. With the drought and honey shortage being experienced nationally the pollen collections have been poor when compared to project expectations. However, as a speaker at 3 Beekeeper annual general meetings (NSW, VIC & SA) over the next couple of months, the pollen collections should improve once beekeepers further understand the project. The cold storage tests have also commenced.

Project Title	Natural resource database for the South Australian apiary industry
RIRDC Project No.:	DEH-1A
Start Date:	05/07/99
Finish Date:	31/05/02
Researcher:	Dr. Chris Holden
Organisation:	Department of Environment, Heritage and Aboriginal Affairs (SA) Biodiversity Conservation Program Heritage and Biodiversity Division PO Box 1047 ADELAIDE SA 5000
Phone:	(08) 8204 8736
Fax:	(08) 8204 8889
Email:	holden.chris@saugov.sa.gov.au
Objectives	To create a database of floral resource information for the South Australian apiary industry, which includes details of: 1. the floral resource base on which the industry is dependent, including the reliance of the industry on native vegetation and the most valuable and reliable floral species for honeybees; 2. the distribution of various native plant species as they relate to beekeeping; 3. the frequency with which these resources are used, the land tenure on which they currently exist, and the relative values for honey and pollen as they relate to honeybee nutritional requirements and honey production; and 4. estimates of the value of currently used apiary sites on both private and Crown lands in terms of dollar value or as a percentage of the State annual production.
Current Progress	The researcher Dr David Paton has employed research assistant Emma Crossfield to compile the report. Subsequent to the poor response to the initial questionnaire (sent to 237 registered beekeepers), information was distributed through the South Australian Apiarists Association newsletter, meetings arranged with small collections of beekeepers, a simpler form distributed to non-respondents, and finally phone-calls to those beekeepers who had still not responded. Of the beekeepers approached, 50% have responded. This accounts for 30% of the hives in South Australia and suggests a higher proportion of smaller operators had responded. At best, another 20% of beekeepers may provide information. At least 10% and perhaps as high as 30% of South Australian beekeepers refuse to provide information. Map production will be finalised in May 2002. This project is on track for publication in June 2002.

Rural Industries Research & Development Corporation

Rural Industries
Research & Development Corporation