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OIE Reference Laboratory Reports
Activities in 2010
Name of disease (or topic) for
which you are a designated OIE
Reference Laboratory:
Address of laboratory
Bee diseases
Chemisches Veterinäruntersuchungsamt Freiburg
P.O.B. 100462, D-79123 Freiburg, Germany
Tel.:
(+49-761) 150.21.75
Fax:
(+49-761) 150.22.99
e-mail address:
[email protected]
website:
Name of Head of Laboratory
(Responsible Official):
Dr Wolfgang Ritter
Name of OIE Reference Expert:
Dr Wolfgang Ritter
Name of writer of this report
(if different from above):
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
1
Bee diseases
Part I: Summary of general activities related to the disease
1.
Test(s) in use/or available for the specified disease/topic at your laboratory
Disease
Test
Positive
Negative
Total
Acariosis
Microscopic
1
87
88
American foulbrood
Culture tech.
264
1428
1692
RT-PCR1
30
10
40
European foulbrood
Culture tech
16
13
29
Nosemosis
Microscopic
29
64
93
Varroosis
Macroscopic
115
16
131
Tropilaelaps spp.
Macroscopic
0
115
115
Small hive beatle
Macroscopic
0
112
112
Acute Bee Paralysis Virus (ABPV)
RT-PCR2
16
84
100
Chronical Bee Paralysis Virus
(CBPV)
RT-PCR1
0
12
12
Deformed wing virus (DWV)
RT-PCR2
68
32
100
Kashmir bee virus (KBV)
RT-PCR1, 2
0
10
10
Sac brood virus (SBV)
RT-PCR1, 2
6
94
100
1Complete
body; 2Heads only
Diagnosis of bee diseases refers to parasites, bacteria, fungi and viruses as agents
As parasites are recognised the ectoparasitic mites Varroa destructor and Acarapis woodi as well as the
microsporidium Nosema spp. All of them are diagnosed exclusively by macroscopic or microscopic examinations
and identified on the basis of their morphological qualities. Bacterial diseases mainly occur in the bee brood.
Paenibacillus larvae (formerly called Bacillus larvae), the agent of American Foulbrood, and Melissococcus
pluton, the agent of European Foulbrood, are cultivated on special nutrient media and then identified under the
microscope. After being cultivated on a nutrient medium, the agents of the most important fungus diseases,
Ascophaera apis and Aspergillus flavus, can easily be distinguished by microscopy. Some of the agents can be
analyzed by PCR technique. Bee viruses can be identified by PCR. The respective tests were performed in the
frequency shown in the table.
2.
Production and distribution of diagnostic reagents
For the diagnosis of European Foulbrood and American Foulbrood, strains of P.l. and M.p., respectively, were
given to different diagnostic laboratories as reference strains.
On the national level another institute in our order produces and distributes standards for detection of P.l. in
honey.
The diagnosis of P.l. and various viruses was performed for different countries.
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Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
Bee diseases
Part II: Activities specifically related to the mandate
of OIE Reference Laboratories
3.
International harmonisation and standardisation of methods for diagnostic testing or the
production and testing of vaccines
The CVUA Freiburg as national reference laboratory for notifiable bee diseases continued the interlaboratory test
with further testing institutions in different countries. An international interlaboratory test for different OIE
members has been prepared for the year 2011.
4.
Preparation and supply of international reference standards for diagnostic tests or vaccines
In cooperation with other institutes different reference materials were created.
5.
Research and development of new procedures for diagnosis and control
Paenibacillus larvae bacteremia in humans
In a collaborative research program with the University Hospital Freiburg, Germany, P. larvae was detected in
sterile compartments of 5 patients with clinical and laboratory evidence of infection. Given the fact that all patients
were injection drug users, the mode of infection was thought to be intravenous administration of contaminated
methadone, resulting in P. larvae bacteremia. Our hypothesis is supported by the isolation of P. larvae from honey
or honey-containing methadone provided to 2 patients. Recently, several Paenibacillus species have been reported
to cause bacteremic infections in humans. Several aspects provide strong evidence for a genuine P. larvae
bacteremia. First, the present cases were observed over a period of several years, and detection of P. larvae thus
occurred in different charges of blood culture bottles, which argues against pseudobacteremia. Second, isolation of
P. larvae was reported independently by two microbiology laboratories, making contamination highly unlikely.
Third, in patient 1 isolation succeeded at different times and in samples of different compartments. Moreover, the
detection of P. larvae in honey-prepared methadone and honey strongly suggests genuine bacteremia as a
consequence of injection of contaminated material. Biochemical and molecular identification of P. larvae may be
difficult and time-consuming. Misinterpretation of blood culture results because of incomplete differentiation or
confusion with other gram-positive spore forming-bacteria (e.g., Bacillus species) has to be taken into
consideration. Underestimation of the frequency of true P. larvae bacteremia therefore cannot be excluded. Thus,
infectious disease physicians, microbiologists, and pharmacists need to be aware that injection of material
contaminated with P. larvae, such as honey-prepared methadone, may cause bacteremic infection.
Diagnosis of Small Hive Beatle
In 2010, the year under review, diagnosing methods of different diseases were verified. The method for diagnosing
the Small Hive Beetle in the bee colony developed by us (Schäfer et al, 2009) proved to be efficient and easily
applicable, both for veterinarians and animal owners. Therefore, this method is recommended for general practice.
Diagnosis of Paenibacillus larvae in Debris of bee colonies
The examinations on the evidence of Paenibacillus larvae, the cause of American Foulbrood, were extended to the
analysis of faeces of bee colonies. A method was developed at the Institute in Dole (CSR). According to first
results, this method could be suitable especially for the examination of bee colonies in winter cluster. A final
conclusion, however, will only be possible after termination and evaluation of the complete field test next year.
Pathogenesis of European Foulbrood in Germany and Switzerland
The examinations on different variations of the pathogenesis of European Foulbrood have been continued.
Whereas this disease has become more and more epidemic in Switzerland, it is regionally restricted in Germany
and only rarely spread within neighboring apiaries. As flight routes of the bees from both areas are overlapping,
different types of pathogens can be excluded as a reason. According to the results achieved until now,
management method and hive types could have an influence on the epidemic process. The examinations are going
to be continued and intensified, especially along the borderline of both countries.
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
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Bee diseases
6.
Collection, analysis and dissemination of epizootiological data relevant to international disease
control
Data on bee losses in the EU and other parts of the world were collected.
7.
Provision of consultant expertise to OIE or to OIE Members
The OIE expert Dr W. Ritter participated at various hearings of experts at the Veterinary Board of the European
Union.
8.
Provision of scientific and technical training to personnel from other OIE Members
The OIE expert, Dr Ritter, participated in the establishment of diagnostic methods at various laboratories.
9.
Provision of diagnostic testing facilities to other OIE Members
The Service was requested from Austria, Italy, Switzerland, Czech Republic and Turkey on an individual level
(institutes, companies, organisations).
10. Organisation of international scientific meetings on behalf of OIE or other international bodies
In 2010, the year under review, the 42 th International Congress of Apimondia at Buenos Aires (Argentine) was
prepared. More than 4000 participants are expected in harvest 2011. Among other topics, a total number of four
symposiums are planned about different topics of bee health. In the week before the congress, an OIE symposium
will be held. The diagnosis and control of Varoosis and the American Foulbrood will be the focal points. They are
organized in cooperation with local scientists and collaborators of the veterinary authorities.
In cooperation with an NGO and by order of the European Commission, the author has been offering training
courses for veterinarians working in the civil service. During the one week’s course at Prague (Czech Republic)
the basic facts about diagnosis and control of the most important bee diseases as well as the most important
official and international regulations are taught. In the afternoons, staff members of the Czech Institute at Dole
will demonstrate the contexts in practice. During the year under review, two courses with 45 participants each
from different countries of the European Union and neighboring countries were realized. Another two courses are
planned for the following year.
11. Participation in international scientific collaborative studies
COLOSS/COST Action
As official delegate of the Federal Republic of Germany, the author participated at different activities of the
Colloss Project funded by the European Union (COST) since 2009.
12. Publication and dissemination of information relevant to the work of OIE (including list of
scientific publications, internet publishing activities, presentations at international conferences)

Presentations at international conferences and meetings
Ritter W. Different statements and presentations on bee losses at different meetings.
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
Scientific publications in peer-reviewed journals

Conte Y., Marion E., Ritter W. Varroa mites and honey bee health: can Varroa explain part of the colony
losses? Apidologie 41 (2010) 3353-363*
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
Bee diseases

Rieg S., Bauer T.M., Peyerl-Hoffmann G., Held J., Ritter W., Wagner D., Kern W.V., Serr A. 2010.
Paenibacillus larvae Bacteremia in Injection Drug Users. Emerging Infectious Diseases (16) 3: 487-489

Genersch E., von der Ohe W., Kaatz H., Schroeder A., Otten Ch., Büchler R., Berg S., Ritter W., Mühlen W.,
Gsider S., Meixner M. Liebig G., Rosenkranz P. 2010. The German bee monitporing project: a long term
study to understand periodically high wintrer losses of honey bee colonies. Apidologie: 41:332-352.

Schaefer, M., Ritter, W., Pettis, J.S., Teal, P.E., Neumann, P. 2010. The role of infestations with Aethina
tumida and Varroa destructor. J. Econ. Entomol. 1o3 (1):10-16

Other communications
Several statements, expert opinion and consultancy reports
13. Inscription of diagnostic kits on the OIE Register
i)
Did you participate in expert panels for the validation of candidate kits for inscription on the
OIE Register? If yes, for which kits?
None
ii)
Did you submit to the OIE candidate kits for inscription on the OIE Register? If yes, for
which kits?
None
_______________
Annual reports of OIE Reference Laboratories and Collaborating Centres, 2010
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