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PCR testing for selected diseases of bees
GREGOROVÁ, Sabina
Nosema ceranae is a species of microsporidia that negatively affects Apis mellifera. It is most commonly transmitted by coprophagy, or eating faeces when filling faecal sacs. It is a worldwide pathogen that is considered to be one of the causes of CCD (colony collapse disorder). The lesser known Crithidia mellificae belongs to the class Trypanosomatidae, which acts similarly like N. ceranae. Both of these pathogens target the guts of honey bees. As a result, infected bees weaken and the colony dies. The aim of this bachelor thesis was the practical examination of selected Apis mellifera samples from the beehive of the University of South Bohemia Faculty of Agriculture and Technology using the PCR method. In the theoretical part I describe Nosema ceranae and Crithidii mellificae and their effect on honey bees. Furthermore, the route of transmission of these pathogens and the possible treatment of infected honey bee colonies are mentioned. In the practical part, which I did in the laboratory at the Department of Plant Production of the Faculty of Agriculture and Technology, the procedure of DNA isolation, PCR and gel electrophoresis is described in detail. The evaluation of the results is also presented in this part.
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Lotmaria passim and Crithidia mellificae prevalence in bee colonies in Czechia
VACÍKOVÁ, Kristýna
In the Czech Republic, beekeeping has a traditional position among human economic activities. Honey bees have been bred by humans for about 15,000 years. In recent years, increased losses of farmed bee colonies have attracted the attention of scientists and the public. A number of biological and abiotic factors play an important role in the loss of bee colonies, especially the action of pathogens and parasites. Trypanosomatidae is one of the most numerous groups of parasitic protozoa. They are unicellular parasites and can be hosted by vertebrates, including humans, insect and plants. The aim of may bachelor thesis was to master the methodology of analysis of the occurrence of trypanosome parasites Crithidia mellificae and Lotmaria passim in the digestive tract of bees using PCR, including DNA isolation and gel electrophoresis .
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Molekulární epidemiologie druhů Crithidia mellificae a Lotmaria passim v populaci včelstev
VOČADLOVÁ, Kateřina
The increased honey bee colony losses in the last decade gain a considerable attention of scientists and public. The causes of these losses include a wide range of biotic and abiotic factors, but pathogens and parasites are probably the main ones. Monoxenous trypanosomatids in honey bee gut Lotmaria passim and Crithidia mellificae were neglected for a long time but according to recent studies they seem to participate in those colony declines. Trypanosomatids are widespread parasites, including also the causes of some human illnesses, such as sleeping sickness, Chagas disease and leishmaniasis. The mechanism of the effect on honey bee health is not well understood so far. The aim of this thesis was to verify the occurrence of the trypanosomatids in honey bee samples from two regions in Czech Republic. The methods, based on detection of specific DNA loci, confirm the high prevalence of L. passim, which was founded in majority (71 %) of the samples. On the contrary, no samples were positive to C. mellificae.
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The parasitic microorganisms in immunodeficient and healthy population of honebees (Apis mellifra)
Bičianová, Martina ; Hubert, Jan (advisor) ; Titěra, Dalibor (referee)
Immunodeficient honey bee (Apis mellifera) colonies suffer from broad range of parasites including eukaryotic protozoa. Despite this fact, the eukaryotic parasites are still poorly documented in the Czech Republic. The presence of eukaryotic parasites (Nosema ceranae, Nosema apis, Crithidia mellificae and Apicystis bombi) was observed in different apiaries in the Czech Republic. The samples were taken in 9 apiaries in 53 beehives during the 2014/2015 season. From each beehive, 10 adult of honey bees were taken from the peripheral comb in triplicate. DNA was isolated from every sample of honey bees. The parasites were detected by polymerase chain reaction (PCR) with specific primers. The treatment fall of parasitic mite Varroa destructor was obtained from beekeepers for season of 2014. Crithidia mellificae was detected by 5 types of specific primers (SEF, SER; SSU, SSU rRNA, Cyt b, Tryp cyt b) and positive amplicons were cloned and sequenced. The obtained sequences were compared with GeneBank and showed similarity from 98-100% to sequences of Lotmaria passim (Trypanosomatid). Crithidia mellificae was not detected. L. passim had prevalence of 79,2% and is reported in the Czech Republic for the first time. Primer Tryp-cyt b is recommended for the routine detection of L. passim. Nosema ceranae was...
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