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Genetické markery pro druhovou identifikaci a studium genetické variability forenzně významných druhů masařek (Sarcophagidae).
JÁNOVÁ, Nicole
The aim of this thesis was to select and test molecular markers for species identification and detection of genetic variability of forensically important Flesh flies that are most common in the Czech Republic (Sarcophaga variegata, africa, noverca, carnaria, albiceps, incisilobata, lehmanni, argyrostoma, caerulescens). Both mitochondrial (COI) and nuclear (ITS, EPIC) markers were successfully amplified and sequenced for this purpose. Bacteria Wolbachia was detected using the 16S marker.
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Forenzně významní zástupci řádu brouci (Coleoptera) vyskytující se na území České republiky
Wrzecionková, Nicole Elizabeth
The process of gradual settlement of the dead body by necrophagous insects plays an important role in forensic practice, especially when determining the time of death. This work deals with the decomposition of dead body of vertebrates, forensically important representatives from the order of Coleoptera in the Czech Republic. Experimental objects for our purposes were domestic pig and chicken. The research was realized from December 2017 to March 2019. During the study samples of larvae and adults were collected from the dead bodies and subsequently identified by anatomical-morphological and molecular-genetic methods. These methods were compared with each other. For research purposes was necessary to isolate the mitochondrial DNA, amplify the fragment of cytochrom c oxidase I (COI) gene by the PCR method and then sequencing it mitochondrial DNA. The BLAST and BOLD databases showed the same outputs and based on the results, the individuals were classified into individual families of the Coleoptera order.
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Identifikace forenzně významných druhů hmyzu s výskytem v chladných měsících
Skoupá, Kristýna
The identification of insect inhabiting dead bodies is particularly relevant in the area of forensic science, especially for post-mortem interval determination, as evidence of post-mortem manipulation of the body, evidence of the presence of toxic substances, etc. This work deals with the determination of forensically important insects occurring in cold months and extreme conditions based on molecular-genetic analyzes. For this purpose, were established 3 experimental fields in Brno-venkov and Blansko district and at the bottom of the Macocha Abyss with a specific cave microclimate. DNA isolation was performed, amplified the cytochromoxidase I gene and sequenced this part of mitochondrial DNA of insect specimens. Then the sequence was compared to the BLAST and BOLD databases, the BOLD showing a higher percentage of matching. This way, were identified 24 insect species from a total of 13 different families. The most of the trapped insects formed Calliphora vicina and Calliphora vomitoria, typical representatives of winter necrophagous fauna.
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Molekulární markery pro detekci genetické variability přírodních populací forenzně významných druhů bzučivkovitých (Calliphoridae, Diptera)
KLOJDOVÁ, Martina
This thesis was focused on suitability of selected molecular markers for detection of genetic variability of four species from the family Calliphoridae important in the forensic entomology. A set of eight markers, both mitochondrial (COI, ND6, CytB, CR) and nuclear (RP S12, RP S13, RP L12, PB2), were applied on samples from natural populations of representative species (Calliphora vicina, Lucilia caesar, Lucilia sericata, Phormia regina) most common in the Czech Republic. Level of detected variability was evaluated and compared, both with respect to the particular species as well as their geographic origin.
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Aplikace molekulárních metod na identifikaci nekrofágních zástupců řádu Diptera, typických pro Jihomoravský kraj
Mifková, Tamara
Necrophagous insects plays an important role, especially in forensic practice, especially in determining the time of death. This work was aimed to monitor necrophagous species of the Diptera order in selected localities of South Moravian Region - Rakvice and Sokolnice. Furthermore, these necrophagous species were identified with anatomical-morphological and molecular genetic methods, which have been compared to each other. For this purpose it was necessary to isolate a DNA segment and amplify cytochrome oxidase I (COI) gene by the PCR method and with further processing to obtain the sequence of selected individuals from the mitochondrial genome. The results of the experiment more coincided with morphological identification database BOLD outputs than with outputs from the BLAST database. Anatomical and morphological identification cannot always accurately determine the genus and species necrophags, the assesment is not dependent not only on the development stage of the insect and its condition, but also on the expertise of the determinator. Most accurater results are achieved with use the combination of anatomical and morphological and molecular-genetic methods of determination, which is confirmed by the results of this thesis.
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The effect of previous freezing of cadavers on the following colonization by flies of the family Calliphoridae
Černá, Monika ; Barták, Miroslav (advisor) ; Olekšáková, Tereza (referee)
The aim of the thesis was to rally available information and draw up a review of literature data on the topic of how freezing of a carcase or its storage at low temperatures affects its following colonization by insects while open exposition.
Forensic entomology is one of the special and ever-developing field of criminology. It uses a knowledge of the biology, ecology and especially the life cycles of insect, mainly flies and beetles, to assist in the investigation of violent crimes in estimating of post mortem interval (PMI) or determining the cause of death. Insect colonization openly exposed corpses go through 5 to 8 successional waves depending on the surrounding environment. One of the important factors is the temperature which affects the decomposition process, where decomposition processes are longer at lower temperatures and on the contrary at shorter at higher temperatures. Temperature has a similar effect on development of insect, which develops only within the effective temperatures, whose values vary by species.
The study performs an experiment which should verify whether insect is able to oviposit on the frozen carcasses and whether pre-frozen carcasses slow the subsequent colonization by flies of the family Calliphoridae. During the experiments, a total of twelve pieces of domestic fowl (Gallus gallus f. domestica L.), weighing about 1.5 kg, were sacrificed in the same way, and a half of them was frozen before the exposition. Experiment results showed that the percentage of each species in both variants differed. Frozen and fresh cadaveres differed in the total nu mber of individuals (larvae), which developed on them, and the way in which the dead bodies decayed.
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The influence of height above ground on colonization of carcasses by blow flies (Calliphoridae)
Mudrochová, Michaela ; Barták, Miroslav (advisor) ; Olekšáková, Tereza (referee)
The forensic entomology is a special discipline of criminology which uses knowledge about insects cycles, mainly Diptera and Coeloptera, to determine the post-mortem interval. The post-mortem interval is time from death to finding of a corpse.
In the Czech Republic, there wasn´t implemented any terraneous experiment that would prove the truth or disprove about the influence of height above ground on colonization of carcasses by blow flies (Calliphoridae). Experimental part was directly focused on that. For the experiment, there was used a building with six floors, precisely it was used balconies on the third and sixth floors and bush at the ground level in the shadow in front of the building. Meat baits which represented cadavers were on the all floors. Baits were used for colonization and breeding species of flies which were determined afterwards. Finally, obtained data were evaluated for their utilization in forensic entomology.
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Ecological and evolution strategies of necrophagous beetles (Coleoptera)
Jakubec, Pavel ; Růžička, Jan (advisor) ; Petr, Petr (referee)
Necrophagous beetles (Coleoptera) are very interesting and diverse ecological group of species with an immense impact on a natural nutrient cycle. Their main food source and breeding ground are carrions of vertebrates, human remains included. This relationship is often used in various ways by forensic entomology, but its potential was not jet fully reached, because our knowledge of biology and ecology of these beetles is very much incomplete. In this thesis I would like to explore geographic distribution, ecological requirements and developmental biology of several Central European necrophagous beetles as an outcome of their ecological and evolution strategies. For that I raised three broad research questions. Which factors are determining the local abundance of carrion beetles (Coleoptera: Silphidae)? How the current geographical distribution of open-landscape carrion beetles looks like in the Czech Republic? How the temperature affects the development of Sciodrepoides watsoni (Spence, 1813)? We found out that soil type can have significant effect on abundance of carrion beetles. They showed preference for chernozem -- Nicrophorus antennatus (Reitter), N. germanicus (Linnaeus), N. interruptus (Stephens), N. sepultor (Charpentier), Silpha obscura obscura (Herbst), T. sinuatus (Fabricius) or for fluvisol as did N. humator (Gleditsch). These findings support our hypothesis that soil type could be an important factor determining the occurrence of necrophagous European carrion beetles. To collect novel data of the current geographical distribution of carrion beetles we used 420 baited pitfall traps at 84 localities, and we collected 71 234 specimens of 15 silphid species. Among them, three endangered carrion beetle species listed on the Czech Red List of Invertebrates, were found. Two are vulnerable thermophilic species of open landscapes, Nicrophorus antennatus (Reitter, 1884) (collected around Louny and Židlochovice) and Nicrophorus germanicus (Linnaeus, 1758) (Louny, Zábřeh and Židlochovice). The third is the near threatened species, Nicrophorus sepultor Charpentier, 1825 (collected around Louny, Kutná Hora, Zábřeh and Židlochovice), which also prefers open landscapes. We studied development of common Holarctic beetle Sciodrepoides watsoni under five constant temperature regimes in laboratory (15, 18, 21, 25 and 28°C). Parameters of thermal summation models and their standard errors were calculated for each developmental stage (egg, three larval instars and pupae). We also find a new character for larval instar determination (head width) and proposed novel approach for future studies of size-based characters in instar determination.
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Species composition of invertebrates during cadavers colonizatin in shallow graves
Pikal, Lukáš ; Červený, Jaroslav (advisor) ; Hana, Hana (referee)
In this work, I deal with forensic entomology and representation of invertebrates species which colonise carcasses found in shallow graves, covered with a layer of soil. There was a field experiment conducted from mid-May 2015 to mid-September 2015 for the purpose of this topic. It took place in the Bohemian Forest, known in Czech as Šumava, in the Kundratice village, where shallow graves were dug in the garden of a family house. There were 12 graves in total. There was a carcass of domestic fowl (Gallus gallus f. domestica, Linné, 1758), each weighing about 2 kg, inserted into each grave and then buried. The distance between the graves was 1.5 m. All the individuals of domestic fowl were killed in the same way under the same conditions. The individual graves were divided into four sectors, each sector containing graves with depths of 10 cm, 30 cm, and 50 cm. Subsequently, one sector was dug up every month, and all developmental stages of insects were picked up from each depth, and the samples were then sorted according to whether they were found in the soil or on the carcase itself. The identified samples were then determined by a specialist. Finally, the findings were compared with literary sources and either refuted or confirmed. The experiment has verified the knowledge that members of the Calliphoridae family do not participate on the decomposing process of buried carcasses.
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Family Muscidae on large cadavers with the beginning of decomposition in the winter and its use in forensic practice
Dočekalová, Tereza ; Barták, Miroslav (advisor) ; Klimešová, Vanda (referee)
Forensic entomology is a one of the special fields of criminology. That uses knowledge of biology, ecology and especially insects and other invertebrates, which occur on cadavers or in its vicinity. These and other arthropods are used as circumstantial evidences during the investigation and also verify other evidence in civil and criminal law.
This bachelor thesis deals with the use of Diptera of forensic practice with focusing
on family Muscidae by form of a scientific work. The first part of the thesis is focuses
on history and current use of forensic entomology, advances in characterization of thematic cases in which the family Muscidae was used as well as describes main species of the family. The second part describes the field experiment in which the presence of the family Muscidae on model organism (pig carrion) was studied and its use in forensic practice.
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