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Study of physiology and searching of proteomic tools useful in suppression and detection of Varroa destructor
Holenková, Martina ; Erban, Tomáš (advisor) ; Titěra, Dalibor (referee)
This work is focused on the study of physiology and proteome of the the mite Varroa destructor and on comparison with the honeybee Apis mellifera. Varroa is currently a major problem for beekeeping, because infects most of the colonies. The control of the mite can not be done without residues both in the hive and for example in the honey or other resources used by the man. Another problem can also be the simultaneously discussed issue of the connection with the Colony Collapse Disorder. The internal anatomy of V. destructor was studied by using paraffin histology. On histological sections stained using hematoxylin and PAS was possible to observe the mite digestive system, but also egg, ovaries or brain. The primary aim of this study was to identify the proteins of mite V. destructor and bee A. mellifera as a host of this parasite. For the electrophoretic separation was used two- dimensional gel electrophoresis, where the second dimension was carried out using 12% and 15% SDS-PAGE. The most abundant spots were selected for analysis using MALDI TOF/TOF mass spectrometry. The most abundant protein identified in samples of V. destructor was hexamerin, arginine kinase or hemelipoglycoprotein precursor. Hexamerins were also identified as the major proteins in the pupae samples. On the contrary the main...
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Proteolytic Enzymes of Vegetative Forms and Spores of the Bacterium Paenibacillus larvae
Hrabák, Jaroslav ; Martínek, Karel (advisor) ; Kotyza, Jaromír (referee) ; Titěra, Dalibor (referee)
Due to the high resistance of the spores, the bacterium Paenibacillus larvae is the most dangerous bacterial pathogen of the honey bee (Apis mellifera). Thanks to its biological properties and restricted pathogenicity, this bacterium can be used as a model organism to study gram positive sporulating aerobic rods. This work is focused on completing information about secreted proteases of this bacterium and in a study of proteases bound in a spore structure. MYPGP medium was used for the cultivation of P. larvae. In this medium, lysis of the culture was shown after 40 hours of cultivation. The pH of the medium decreased below 6.4 by lysis. The induction of temperate bacteriophage BLA was detected as a causative agent of this lysis. A new sporulation medium called HCBB agar was proposed for the sporulation of P. larvae. In comparison with HCBB agar with MYPGP agar by 31 strains of P. larvae stored in our collection, HCBB agar was evaluated as an appropriate sporulation medium with a median of sporulatin 4.2 ' 106 spores per cm2 in aerobic conditions and 5.65 ' 106 spores per cm2 in aerobic conditions with 10 % CO2. For purification of the secreted proteases, a one-day culture incubated at room temperature was used. Optimal purification of 87/74 kDa and 42/40 kDa proteases was observed after application of this...
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Melissococcus plutonius causing European foulbrood
Navrátilová, Blanka ; Hubert, Jan (advisor) ; Titěra, Dalibor (referee)
European foulbrood is a bacterial disease attacking honey bee larvae worldwide. It is caused by bacterium Melissococcus plutonius, which is a non-spore-forming, Gram positive, anaerobic bacterium. The adult bees are not affected but serve as a vector of the disease as they carry the bacterium within their own bodies and can travel big distances from their hive and also may interact with other hives especially when their own colony is suffering. Once the bacterium is introduced into the colony, it either remains benign and unnoticed for years, keeping its population low, or it can multiply vigorously within the brood and destroy the entire bee populations. Despite having been described many decades ago, M. plutonius as such along with its virulence remain poorly understood and therefore there is no treatment efficient enough that would keep this bacterium along with the disease under control. Hence it is of a great importance to recognize its presence soon enough to prevent the outbreak. This thesis brings together the knowledge we have so far about this mysterious bacterium and sums up how European foulbrood is being treated all around the world.
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A new approach for assessing the suspected pesticide poisonings of honeybees: certified methodology
Erban, Tomáš ; Kamler, Martin ; Kadlíková, Klára ; Markovič, Martin ; Titěra, Dalibor ; Seifrtová, Marcela ; Halešová, Taťána
Honeybee colonies are exposed to hundreds of plant protection products, which are especially due to improper use possible source of honeybee poisoning. The methodology introduces to the issue of honeybee poisoning including legislation and contributes with novel aspects to the indication and evaluation of the suspected honeybee poisonings, which are anually investigated by the State Veterinary Administration and the Central Institute for Supervising and Testing in Agriculture in Czechia. The methodology provides instructions to identify honeybee posining and notification of that finding. Moreover, possible confusions of the poisonings with honeybee diseases are highlighted. Some sections of the methodology provide innovative recommendations for improvement of honeybee poisoning assessing including determination of the extent of colony contamination. The present methodology is useful for the state administration, beekeepers, scientific and educational purposes.
Fulltext:  PDF
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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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Study of physiology and searching of proteomic tools useful in suppression and detection of Varroa destructor
Holenková, Martina ; Erban, Tomáš (advisor) ; Titěra, Dalibor (referee)
This work is focused on the study of physiology and proteome of the the mite Varroa destructor and on comparison with the honeybee Apis mellifera. Varroa is currently a major problem for beekeeping, because infects most of the colonies. The control of the mite can not be done without residues both in the hive and for example in the honey or other resources used by the man. Another problem can also be the simultaneously discussed issue of the connection with the Colony Collapse Disorder. The internal anatomy of V. destructor was studied by using paraffin histology. On histological sections stained using hematoxylin and PAS was possible to observe the mite digestive system, but also egg, ovaries or brain. The primary aim of this study was to identify the proteins of mite V. destructor and bee A. mellifera as a host of this parasite. For the electrophoretic separation was used two- dimensional gel electrophoresis, where the second dimension was carried out using 12% and 15% SDS-PAGE. The most abundant spots were selected for analysis using MALDI TOF/TOF mass spectrometry. The most abundant protein identified in samples of V. destructor was hexamerin, arginine kinase or hemelipoglycoprotein precursor. Hexamerins were also identified as the major proteins in the pupae samples. On the contrary the main...
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Proteolytic Enzymes of Vegetative Forms and Spores of the Bacterium Paenibacillus larvae
Hrabák, Jaroslav ; Martínek, Karel (advisor) ; Kotyza, Jaromír (referee) ; Titěra, Dalibor (referee)
Due to the high resistance of the spores, the bacterium Paenibacillus larvae is the most dangerous bacterial pathogen of the honey bee (Apis mellifera). Thanks to its biological properties and restricted pathogenicity, this bacterium can be used as a model organism to study gram positive sporulating aerobic rods. This work is focused on completing information about secreted proteases of this bacterium and in a study of proteases bound in a spore structure. MYPGP medium was used for the cultivation of P. larvae. In this medium, lysis of the culture was shown after 40 hours of cultivation. The pH of the medium decreased below 6.4 by lysis. The induction of temperate bacteriophage BLA was detected as a causative agent of this lysis. A new sporulation medium called HCBB agar was proposed for the sporulation of P. larvae. In comparison with HCBB agar with MYPGP agar by 31 strains of P. larvae stored in our collection, HCBB agar was evaluated as an appropriate sporulation medium with a median of sporulatin 4.2 ' 106 spores per cm2 in aerobic conditions and 5.65 ' 106 spores per cm2 in aerobic conditions with 10 % CO2. For purification of the secreted proteases, a one-day culture incubated at room temperature was used. Optimal purification of 87/74 kDa and 42/40 kDa proteases was observed after application of this...
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Content of succrose in authentic honey from Czech Republic
Mrázková, Iveta ; Titěra, Dalibor (advisor)
The work is focused on the analysis of data on sugar composition of honey coming from beekeepers in the Czech Republic. The database includes a number of years and several hundred locations. The aim is to test the hypothesis that sucrose content in honeys vary in individual years in connection with the course of the weather, especially the sufficiency or deficiency of rainfall, so that in rare cases it can even unadulterated honeys sucrose content exceed the limit specified standard.
It will formulate recommendations for action at the findings of the high sucrose content of genuine honeys to prevent unauthorized punishment.
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Abundance of honeybee
Brandejsová, Jana ; Titěra, Dalibor (advisor) ; Marcela, Marcela (referee)
Determining place thesis Abundance of bee colonies in the country and the carrying capacity of the land register. Establishing a Connection in Úvaly near Prague with three beekeepers. For these three beekeepers were required to determine reserves of pollen with three visits in the period of May, June and July. Each measurement was carried out always at the three strongest hives. Proven reserves of pollen was performed using photographs of each honeycomb in the hive. After calculating reserves of pollen, to assess the value of honey to the surrounding plants. The conclusion was to determine whether a load for the site Úvaly near Prague is present or not enough bees and suggest possible solutions.
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Beekeeping farms in the micro-region Holicko
Ropek, Milan ; Titěra, Dalibor (advisor) ; Hana, Hana (referee)
This Diploma Thesis is focused on obtaining data, how many bee colonies can be found in the micro-region Holicko. Then it wants to confirm or disprove the hypothesis, if the number of bee colonies is sufficient in this micro-region.
To meet this objective the analysis in this area was made. The list of bee colony territories was written in collaboration with ČSV, especially with the aid of ČSV ZO Holice. Maps of bee colonies in particular areas of the micro-region Holicko were made according to the data. There you can see flight circuit of bee colonies and their overlapping in relation with bee food sources.
The bees have really important role in the nature and an irreplaceable function for men. They provide known bee products and ensure lives of many plant species by pollinating. In the monitored area the number of bee colonies is sufficient but its allocation is not optimal. It means, that there are too many bee colonies in some areas and it can have a negative impact on bee nourishment and their state of health.
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