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Characterization of bifidobacteria and lactobacilli isolated from intestinal tract of wild pigs
Šimurka, Julius ; Rada, Vojtěch (advisor) ; Švejstil, Roman (referee)
The microbiota of the gastrointestinal tract is considered as "acuqrated organ" located in a host organism. There are many performs of specific functions. The microbiota composition changes during the life of the individual, depending on his environment and nutrition. The process of identification and characterization of the bacteria from the intestinal microbiota is an important prerequisite for understanding the function of this "acuqrated organ".
This work deals with the anatomy and physiology of the digestive tract (DT) of pigs and evolutionary and eating differences of wild and domesticated pig, for understanding the natural environment of bacteria from DT, which in turn affects the bacterial diversity of the intestinal microbiota. Also mentioned is the process of domestication, which influenced the present form of domestic pig. The work includes the characteristics of lactobacilli and bifidobacteria, which contains the descriptions, taxonomy, metabolism and habitats. The work also shows the insulating media used in practice for the isolation of bifidobacteria and lactobacilli from DT of pigs and methods of their identification and related methods of characterization.
The available literature confirms the differences in the composition of microbiota DT between wild and domesticated pig, that correlate with other studies which study microbiota diversity of other domesticated and wild animals. The diversity studies of humans microbiota, also confirms the broad spectrum of bacterial species in primitive tribes of humans compared with urbanized man.
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Microbiology of unfiltered beers from craft breweries and the possibility of shelf life extending
Janíková, Jolana ; Rada, Vojtěch (advisor) ; Chládek, Ladislav (referee)
In the last ten years can be recorded a multiple number of minibreweries, called sometimes craft breweries I tis not caused only by demand of kinds of beer characterised by special types of beer styles, but as well by the fact, that they are not anyway regulated
and keep their natural taste and smell. The problém of unregulated (unfiltered
and unpasteurized) beers starts by the contamination during their production and it may cause their relatively fast spoiling. Shelf life of these beers is e few weeks only. The time of their minimal shelf life is influenced by a lot of factors content of alcohol, rest extract
and mentioned extent of contamination. The breweries called as industrial or commercional treat their beers by pasteurization or filtration which cause inactivation or killing of the microorganisms or their elimination. These beers are microbiologically stabile, they are not liable to quick spiling and i tis possible to insure their longer shelf life. Pasteurized a filterd beers show changes in their sensorial character.
The aim of this diploma thesis was to determinate the condition of flash pasteurization of unfiltered beers with the aim of microbiological stability and simultaneously no changes
of their sensorial quality. The experiment is finally in treatment unfiltered beer by differnt dosing of pasteurization units and following microbiological analysis, able to demonstrate
the stability effect. The analysis were used with unfiltered and unpasteurized beers
and with unfiltered beers where the pasteurization doses 20 PU, 30 PU, 40 PU, 60 PU, 80 PU and 100 PU were used. The idividual parameters were watched within the horizont
of 4 months. The part of these tests was even microbiological analysis of the rinse water.
Another task was to find out the dates from the microbiological controls of beer
in selected small breweries, which use in order to heighten of microbiological stability, different kinds of filtration, to verify their effectivity and to appreciate their acceptability
for minibreweries.
The test showed the effectiveness of pasteurization of unfiltered beer with the rations 40 PU minimally. Where higher ratins were used could be seen apparent sensorial changes. The pasteurization of unfiltered beers are for craft breweries useful only in case of higher production of bottled beer or beer for export, because the financial expenses bound
with provision flash pasteurizer are to big and for minibreweries mostly limiting.
The kieselguhr filtration as well the ultrafiltration are for minibreweries unusefull, as they could not be able to insure such conditions that provide their proper affectiveness.
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Molecular genetic characterization of vancomycin-resistant enterococci
Bubeníček, Karel ; Rada, Vojtěch (advisor) ; Igor, Igor (referee)
Summary
Objectives and hypothesis: This thesis concerns the study of plasmids of vancomycin-
resistant enterococci isolated from feces of American crows in the years 2012 - 2013 period.
The hypothesis is that, in various environments, there is one or more types of
epidemiologically significant vanA gene-carrying plasmids that are capable of horizontally
spread.
Methods: Based on PFGE method the number and size of plasmids were detected in selected
isolates of vancomycin-resistant E. faecium. Using PCR method the isolates were subjected
to detection of genes of replicases, relaxases and toxin-antitoxin system of plasmid-bound
resistance genes. Using 19 primers were characterized types of Tn1546.
Results: Of the 12 tested vancomycin-resistant isolates of E. faecium the following number
and size of plasmids was proven using PFGE method: 2 isolates contained two plasmids
(17%), 3 isolates contained three plasmids (25 %), 5 isolates contained four plasmids (42 %)
and 2 isolates contained five plasmids (17 %).
All isolates (n = 12) were then subjected to the detection of genes of replicases, relaxases and
toxin-antitoxin system for typing of plasmids from each plasmid families.
RepA_N family of plasmids:
genes characterizing plasmids related to pRUM: rep17 in 11 isolates (92 %),
gene Axe-Txe was detected in 5 isolates (42 %)
genes characterizing plasmids related to pLG1: rep20 in 7 isolates (58 %)
genes characterizing plasmids related to pAD1: relpAD1 gene was detected in one isolate (8 %)
Inc18 family of plasmids:
genes characterizing plasmids related to pIL501: rep1 gene detected in one case (8 %)
genes characterizing plasmids related to pRES25: rep2 gene in 2 isolates (17 %)
genes characterizing plasmids related to pEF1: relpEF1 detected in 11 isolates (92 %)
pHTB family of plasmids:
genes characterizing plasmids related to pHTB: rep22 gene was detected in 4 isolates (33%) and in 2 isolates gene relpHTB was detected (17%)
RCR family of plasmids:
genes characterizing plasmids related to pRI: positive detection of Rep14 gene in 8 isolates (67%) and in 4 isolates relpRI gene was detected
Small theta-replicating plasmids:
genes characterizing plasmids related to pEF418 plasmids: rep18a gene in 2 isolates (17%)
genes characterizing plasmids related to pB82: rep18b gene was detected in one isolate (8%)
genes characterizing plasmids related to pCIZ2: relpCIZ2 gene was detected in 9 isolates tested (75%)
Types of transposon Tn1546
Using the PCR method types of Tn1546 were characterized. In 4 isolates (n = 12; 33 %)
Tn1546 was characterized as a F3 type. In one isolate (8 %) Tn1546 was characterized as a
type F5, in one isolate (8 %) as a type PP-16. In 6 isolates Tn1546 was untypeable. Most
likely these are new, yet unknown types.
Conclusion: This is the first study of plasmids of vancomycin-resistant isolates E. faecium
isolated from feces of American crows. These results emphasize not only a high proportion of
plasmids in individual isolates, but also a high proportion of genes with horizontal transfer.
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Impact of pesticides on the gut microbiota of human
Kočová, Kateřina ; Rada, Vojtěch (advisor) ; Jakub, Jakub (referee)
Pesticides are substances or mixtures of substances used in agriculture against pests (animals, plants and parasitic fungi), who damage the crops, stocks of agricultural products, food and reduce livestock utility or threaten human health.
The thesis consists of two parts. The first (theoretical) part describes composition and function of gut microbiota; pesticides and their classification, environmental transport and effects of these substances on human health, and glyphosate as the active substance of herbicide Roundup, its mechanism of action, environmental fate and proven effects of glyphosate on human.
The aim of second (practical) part of this thesis was to test impact of pesticide Roundup on the gut microorganisms of human. The impact of pesticide was tested in in vitro pure cultures. Roundup was added in concentrations of glyphosate 17 g/L, 1,7 g/L and
0,17 g/L to the complex media and after cultivation the growth of microorganisms was evaluated. The next samples of stool were collected from human volunteers; these were cultivated similarly in the complex media with different concentrations of glyphosate. The total numbers of microorganisms, bifidobacteria, lactobacilli, enterococci and coliformn bacteria were defined after cultivation.
When the pesticide was tested in pure cultures, concentration of 17 g/L glyphosate significantly inhibited total bacterial growth (P < 0.05), at lower concentrations no difference was observed. The cultivated microorganisms from samples of stool demonstrated only that bifidobacteria are sensitive to glyphosate at the highest used concentration of this pesticide compared with the control sample.
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Clostridium butyricum as a probioticum
Veselá, Jana ; Rada, Vojtěch (advisor) ; Pechar, Radko (referee)
Probiotics are live microorganisms that, when administered in adequate amounts, confer a health benefit on the host. Evidence of positive effects of probiotics is rapidly expanding in gastroenterology. Probiotics find use in antibiotic-associated diarrhea, gastroenteritis and lactose intolerance.
Many different strains of bacteria are considered as probiotic, including clostridia. The most used probiotics are Lactobacillus and Bifidobacterium species. But there are also suggestions that some Clostrium butyricum strains could be also probiotic.
Aim of this study was therefore to evaluate the effectiveness of Miya-Gold (a feed additive containing viable spores of probiotic Clostridium butyricum MIYAIRI 588 strain as an active substance) to the improvement of feed conversion, average daily gain and final body weight in the chickens for fattening and also to observe the improvement of the stability of the gut and crop microbiota.
We found statistically significantly higher (P < 0.05) average weights of individuals in Miya-Gold group than in the control group at the day 7, 10, 20 and 49. By the analysis of microbiota we found at the day 42 statistically significantly lower (P < 0.05) amounts of Escherichia coli in the caecum and Escherichia coli and coliform bacteria in the crop and also lower pH in the crop in the Miya-Gold group.
The analysis of short-chain fatty acids was also performed and we found statistically significantly higher (P < 0.05) amount of butyrate in the caecum of Miya-Gold group.
The results indicate the ability of Miya-Gold to affect positively the performance and the intestinal microbiota of broiler chickens.
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Výroba energie z obnovitelných zdrojů jako forma diverzifikace činnosti zemědělského podniku
Rada, Vojtěch
This thesis deals with renewable energy sources with a focus on the biogas plant as a new form of diversification of farm activities. In the theses is characterized renewable energy source of energy, their development and tools for their support in the European Union and the Czech Republic. At the same time provides the legislative framework for the support of renewable energy sources. Describes the development of biogas plants in the Czech Republic, lists the various types of biogas plants, the principles of their operationn and types of purchase of electricity. The work focuses on the characterization and comparison of two technologically different biogas plants for two years, which are operated in farms. These biogass plants compares between each other by analysis of the costs and revenues related to the production of electricity. At the same time, the thesis analyzed the external and internal environment of these biogas plants
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