Violeta Santrac

This current text, we wanted to draw attention to our work in this field of diagnosis of pathogens of bees, and in the second level, invite all those interested in this type of diagnosis to contact us.

Nekliko years ago, at various conferences in Europe there were announcements that to kritidija, Crithidia mellificae protozoa, trypanosomes, could alone or in cohabitation with other pathogens bees have caused losses of honey bee colonies. In Halle in 2012 at the 5th European Conference, EURBEE, attention was drawn work group of authors: Diadant B. Evans and JD. Gauthier L. Neumann P with the headline: “Chritidia mellificae is widespread in Europe and can be used as predictive marker of honeybee colony colony losses”. The authors of the paper stated that the (* even then) in the United States found in bee colonies, but were not able to examine potential pathogens that could cause this could have on the vitality of bee colonies. In various parts of Europe in trisezonalnom sampling kritidije are found in large numbers of samples in Switzerland N 29, N 97 in France but the authors noted, and the fact that in these communities could not speak to talk about any “recognizable and characteristic clinical symptoms.” In conclusion, this study states that “Crithidia mellificae in case of (vital) reasonably healthy communities harder can cause losses, but in the case of weaker communities where there are intensive infectious capacity, the role of this pathogen can have a significant share in the secondary infectious progress, especially in the period of hibernation community. ”

In the Veterinary Institute of the Republic of Serbian “Dr Vaso Butozan” Banja Luka (VIRSVB) is ongoing laboratory surveillance for the presence of protozoa, tripanozomida, Crithidia mellificae that literature appoints as Lotmaria passium in samples of bees.

During the regular diagnostic requires diagnostics nosemosis of the material in the microscopic dark field and targeted search presence Crithidia mellificae. Diagnosis is simple and the “trained eye” morphology with typical gait trypanosomes is sufficient sign of infection with this parasite. In case of a positive microscopic findings it is possible to do the confirmation of findings by cultivation on liquid mediums, specially designed for this pathogen. The Institute currently has a molecular (PCR) diagnosis of this pathogen and which is in the world because of the specificity, sensitivity and robustness commonly used for evidence in bee colonies. Having laboratory capacity and knowledge of basic microbiological requirements that have this kinetoplastida, we wanted to determine the possible presence of Crithidia mellificae and prevalence in tested samples in the apiaries of the Republic of Serbian and BiH.

The first test we did during the 2014 project “Apin”. This year in May, June, July, on samples of healthy and dead bees, which are made ​​with different diagnostic requirements arrived in VIRSVB, we conducted simultaneously and microscopic examinations of the dark-field X 200, with the aim of detecting Crithidia mellifica.

As much as we seriously try to find the cause in our apiaries, because that would be essential in all tested samples did not find the presence of the “pathogen”. We will continue with the examination of the material in order to, if nothing else, then at least with more security and statistical significance, to inform beekeepers and veterinarians on the status of this “new” pathogens in our apiaries.

In May 2015, I was on education in the framework of the “Super B”, in Ghent, Belgium, a workshop on the application of mRNA technology in controlling infections of honeybee colonies different pathogens (viruses, mites). On that occasion I was staying in the laboratory, incidentally, had the opportunity to see and work on the isolation and cultivation. Reference cultures Crithidia mellifica and method in the laboratory used for cultivating and maintaining kritidije helped us in the work in VIRSVB. In our laboratory conditions, we did “optimize” growth medium Crithidia mellifica, made ​​photos pathogens, and videos about the typical movement tripanozomide in pure culture, which on this occasion made ​​available.

Experience with reference kritidije culture, has helped us to be more researchers from VIRSVB familiar with the “new” pathogenic and also to proceed further with the implementation and verification methods for the detection kritidije.

The video was shot in VIRSVB, telephone through the eyepiece of the microscope for dark field microscopy, magnification 200x, the amount of drops of 10μl inoculum.

Video can see HERE .

Figure 1. Microdot Crithidia mellifica