Trichobilharzia regenti is a neurotropic fluke belonging to family Schistosomatidae. Larvae called schistosomula migrate in the definitive hosts (anseriform birds) throuth the central nervous system (CNS) to their final location in nasal mucosa, where they mature and lay eggs. In contrast with that, the infection of accidental mammalian hosts (including human), is often stopped already in the skin immediately after entering the host. However, some schistosomula are able to reach CNS of experimentally infected mice, and survive there temporarily. Reaction to the CNS infection of mice is usually provided by microglia, astrocytes or the other immune cells infiltrated from the hosts blood. Parasite protects itself against the host reaction with its tegument. It does not serve only as mechanical barrier, but also as main secretoric organ that is capable of active immune evasion. Changes within CNS of the vertebrate hosts, caused by migrating schistosomula of T. regenti, were already described by routine histological and immunohistochemical methods. Till now, there was a lack of informations about interactions of immune cells of the host and the tegument of the parasite on ultrastructural level. To fill this gap in knowledge, two different methods were used: (1) imunohistochemistry in light and electron...

In 1971, the algal class Eustigmatophyceae, Stramenopiles, was detached by Hibberd and Leedale from the class Xanthophyceae, Stramenopiles, on the basis of prominent ultrastructural differences in vegetative cells and more importantly in zoospores, such as the presence of big extraplastidial stigma. The class was divided into four families, six genera and twelve species. It seems so far, that there is a deep dichotomy within the class. Eliáš et al. (unpublished) recom- mend to establish two orders based on molecular sequencing of 18S rDNA and rbcL: Eustigmatales (sen- su Eustigmatophyceae described by Hibberd) and Goniochloridales (order containing Goniochloris, Pseu- dostaurastrum and undescribed relatives). The order Eustigmatales is divided into three separate lineages, most likely families: Eustigmataceae (A1), Monodopsidaceae (A2) and a new family Pseudellipsoidiona- ceae (A3). In this study the ultrastructure of 10 eustigmatophycean strains from three separate lineages was investigated. The typical characteristics of this class as chloroplast without a girdle lamella, a reddish glo- bule and lamellate vesicles were found in all strains studied. The appearance of the lamellate vesicles was found to change during the life cycle. My research indicates that other characteristics, formerly...

Oxymonads (Excavata, Preaxostyla) are a group of anaerobic endobiotic flagellates living primarily in guts of xylophagous insects (cockroaches and termites). Some representatives of the genus Monocercomonoides belonging to the morphologically simplest family Polymastigidae have been described also from the guts of vertebrates. Oxymonads are a group of protist in which mitochondrion has not been proven yet. In this work, we have sequenced gene fo SSU rRNA of two strains of Monocercomonoides and performed phylogenetic analysis of oxymonads. Five selected strains Monocercomonoides isolated from different hosts and distantly related on the phylogenetic tree were studied by using light (DIC and protargol staining) and transmission electron microscopy. The aim was to find differences between these strains. We have focused primarily on the morphology of the nucleus, appearance of the endoplasmic reticulum and structure of the fibre R1. We were able to distinguish the strains by the position of karyosome (central and parietal) and the number of microtubules in the R1 fibre (6-12) and we assume that they represent separate species. The phylogenetic position and appearance of the strains NAU3 were so different that it may represent a new genus. Mitochondria or double-membrane bounded organelles have not been...

Description of Trichobilharzia regenti as a new species of nasal bird schistosome in 1998 was only the first step in our knowledge of this extraordinary parasite. Natural definitive hosts of T. regeni are anseriform birds, but infective larvae - cercariae - are able to penetrate also into mammalian hosts including humans. There they are causative agents of hypersensitive skin immune reaction called cercarial dermatitis or swimmer's itch. Contrary to other schistosomes, miracidia of T. regenti hatch directly inside the definitive host tissue. Schistosomula migrate through the nervous system of vertebrates and, together with adult worms, they have predominantly extravascular localization in definitive hosts. Adult worms have a short lifespan and low degree of sexual dimorphism, connected with lower dependence of adult females on long-term contact with males. During the life cycle, T. regenti can be found within three different environments (freshwater, tissue of intermediate molluscan host and tissue of vertebrate host). Each of the seven developmental stages has a different role in the life cycle which corresponds with different organization of various organ systems. The introductory part of the thesis is focused entirely on ontogenetic changes of surface ultrastructure and body musculature of...

Several studies have been carried out to understand reproductive biology of invasive crayfish. Many researchers have described how invasive species establish their population in new habitats, interactions between native and invasive species, their fast spreading, changing of behavior and biology especially reproductive behavior and even genetics. But it seems that there are still lots of questions and gaps which need to be addressed in case of invasive crayfish species. This thesis tried to focus on different aspects of reproduction biology of invasive crayfish. Generally, the normal crayfish reproduction mode is gonochorism. But, it has been showed in the literature that a few species such as Cherax quadricarinatus, Samastacus spinifrons, Parastacus virilastacus and Pacifastacus leniusculus may have different reproduction modes such as hermaphroditism or intersex. Parthenogenesis has been found in Procambarus fallax f. virginalis. Also, apomictic parthenogenesis has been reported in Orconectes limosus. Moreover, there have been several manipulations which directly affect reproduction biology of crayfish under lab conditions. Additionally, hybridization can be expected between Astacus astacus and Astacus leptodactylus, Orconectes rusticus and Orconectes propinquus under natural conditions. Some studies supposed that chemical factors also one of the reasons which could lead to some changes in reproductive system of crayfish. The objective of the Chapter 2 was to report first evidence of intersex in the signal crayfish, Pacifastacus leniusculus (Dana, 1852). That specimen appeared morphologically male but with both male and female genital openings, as well. Spermophores were obtained from vas defertia of this speciemen. Histological study was carried out and it has shown that both spermatophores and oocytes were present. The gonadosomatic index showed that intersex male had a GSI three times more higher than normal males. The ultrastructure of spermatozoa in three cambarid species Cambarus robustus, Orconectes propinquus, Orconectes rusticus were described and compered with eight previously studied species from family of Astacidae, Cambaridae and Parastacidae. In the studied cambarids, the crest-like protrusions in the anterior part of the acrosome is one of the most remarkable differences in case of morphological feature in spermatozoa and can be used for distinguishing the members of Cambaridae. The results of biometrical data showed that the smallest and biggest acrosome sizes in the studied species were in Parastacidae and Astacidae, respectively. The duration of post-mating spermatophore storage as well as the timing and temperature of spawning in two crayfish species (Pacifastacus leniusculus and Astacus astacus) were investigated in the Chapter 4. Seventy-one pairs of adult signal crayfish (Pacifastacus leniusculus) and thirty six pairs of noble crayfish (Astacus astacus) were used for experiment. The results indicated that there was a significant difference (P<0.05) between noble crayfish and signal crayfish in average duration of the post-mating spermatophore storage. The duration of the post-mating spermatophore storage is longer in the noble crayfish with 34.6 ? 1.7 days than the signal crayfish with 3.9 ? 0.5 days. There were also differences in the timing of mating and egg laying between the signal crayfish and noble crayfish. In the population of signal crayfish both mating and egg laying overlapped, but in the population of noble crayfish there was at least two weeks gap between last mating and first egg laying individuals. Water temperature was significantly (P<0.05) higher during mating and egg laying in the signal crayfish than the noble crayfish. The average temperatures for mating in both species were significantly (P<0.05) higher than the temperatures that they utilized for egg laying. As a conclusion more research is needed for better understand of the reproduction strategies in invasive crayfish.

Members of the group Archamoebae are free-living or endobiotic amoeboid flagellates and amoebae. They live in anoxic or microoxic habitats, and their mitochondria have been reduced. They were originally thought to lack mitochondria and represent one of the earliest eukaryotes. However, this hypothesis has been refuted, and now it is evident that the Archamoebae belongs to the lineage Conosa within the supergroup Amoebozoa, together with aerobic slime molds (Macromycetozoa) and variosean amoebae and flagellates. Relatively simple microtubular cytoskeleton is a characteristic feature of Archamoebae. It consists of a single basal body from which a flagellum arises, lateral root, and microtubular cone. Cytoskeleton of aflagellated genera has been completely reduced. About 350 species names of Archamoebae have been created so far. However, most descriptions were based on inadequate morphological features. The identity of numerous species is uncertain, and many of them are likely synonymous. Another problem is a small amount of available molecular data. During our project, we have substantially improved the dataset of DNA sequences of archamoebae. On the basis of molecular and morphological data, we described 13 new species. We showed that genus Rhizomastix belongs to Archamoebae and displays a new type...

The photosynthesis of the aerial roots of epiphytic orchids has been the subject of numerous studies. However, the roots are always evaluated as a homogeneous structure, even though they actually consist of significantly different areas. This work deals for the first time with the possibility of the spatial distribution of photosynthesis between the root layers, specifically the cortex and the stele. A combination of various microscopic techniques, the histochemical characterization of the apoplastic barriers and the immunohistochemical localization of the photosynthetic enzyme phosphoenolpyruvate carboxylase has been used. The results show that well-developed chloroplasts in the stele probably occur in the subfamily Epidendroideae in all epiphytic representatives, while in the subfamily Vanilloideae they occur occasionally. The ultrastructure of chloroplasts from both areas is systematically different, so it is likely that their functions also differ. Apoplastic barriers are prominent in the roots and differentiate early during root development, which can effectively isolate chloroplasts in the stele from the cortex. Chloroplasts also occur in the sclerenchyma cells of the stele, where were identified hitherto unknown pits in cell walls that could provide gas exchange within the stele....

In the nature, crayfish are represented like predators or a part of food base of some fish. Also in free waters they acts as detrivors and are an important element in the nutrient and energy circle in aquatic ecosystem. The crayfish weren't able to avoid negative impacts of anthropogenic activities such as water pollution and devastation of habitats. For these reasons reproduction of the crayfish is necessary in hatcheries and farms. The main aim of my work was to study the reproductive biology of the crayfish focused on the process of mating, the interval between mating and laying eggs, fertilization and morphological structure of the male and female gametes. Among the secondary objectives there was included using of repeated electrostimulation of the crayfish and the possibility of hybrids of the noble crayfish (Astacus astacus) and the narrow-clawed crayfish (Astacus leptodactylus). The observation of the period of reproduction was realized on representativies of the noble crayfish and the narrow-clawed crayfish throughout natural mating in the tanks. The structure of gametes was observed on the samples taken from thenoble crayfish (Astacus astacus), the narrow-clawed crayfish (Astacus leptodactylus), the signal crayfish (Pacifastacus leniusculus), the red swamp crayfish (Procambarus clarkii) and spinycheek crayfish (Orconectes limosus). The experiments were carried out during the autumn season in 2012 and 2013. The results of observations of the reproduction of crayfish made us clear as for the period and intervals for all phases of the process of mating crayfish. For females the noble crayfish and the narrow-clawed crayfish was found oviposition without the presence of males. Their individual components have been described by using ultrastructure of males and female gametes. Repeated electrostimulation showed at the family Astacidae as a suitable tool for collecting large mass of sperm material. Hybridization experiment proved that it can occur to mate the two species of crayfish in areas where they occur simultaneously. These experiments helped to further knowledge of reproductive biology of crayfish.

Forest stands may act as important carbon storage places - sinks, due to carbon allocation into both the plant biomass in the process of photosynthesis and the soil. Enhancement of CO2 concentration affects a whole range of plant physiological processes and, thus, it is necessary to study its effect on photosynthetic apparatus - leaf anatomical structure and chloroplast ultrastructure. The first aim of the Thesis was to evaluate changes in chloroplast ultrastructure of common beech (Fagus sylvatica L.) under the effects of both elevated CO2 concentration and different irradiance. The second aim was to evaluate if the anatomical parameters obtained from the middle part of the leaf are representative for the whole leaf blade. The trees were grown in glass domes at the Bílý Kříž experimental site in the Beskids Mountains (Czech Republic), owned by the CzechGlobe Institute. Leaves were sampled in 2010 from juvenile trees, which were planted in 2005 being 5-year old and cultivated since then in ambient (AC; 390 micromol/mol) and elevated (EC; 700 micromol/mol) CO2 concentrations. The EC effect was recorded to be an increased proportion of starch grains in the chloroplast median section and decreased proportion of of intergranal thylakoids (IGT) while the ratio of granal to intergranal thylakoids...

<p>U radu je ispitana zastupljenost azotofiksirajućih cijanobakterija, ukupnog broja algi i ukupnog broja bakterija u različitim tipovima zemlji&scaron;ta na jedanaest lokaliteta u Vojvodini, od čega se sedam nalaze u za&scaron;tićenim prirodnim dobrima. Ispitana je brojnost u zavisnosti od dubine pedolo&scaron;kog profila, kao i od godi&scaron;njeg doba. Izvr&scaron;ena je izolacija sojeva azotofiksirajućih cijanobakterija, određena njihova taksonomska pripadnost i osnovne citolo&scaron;ke karakteristike. Ispitana je ultrastruktura vegetativnih ćelija, heterocista i spoljnih struktura na ćelijama fimbrije i pili, transmisionim elektronskim mikroskopom. Izvr&scaron;ena je genetička karakterizacija izolovanih sojeva azotofiksirajućih cijanobakterija PCR metodom analizom STRR fragmenata DNA.<br />Brojnost azotofiksirajućih cijanobakterija i ukupna brojnost algi je bila znatno veća kod hidromorfnih i halomorfnih zemlji&scaron;ta, nego kod automorfnih zemlji&scaron;ta. Najveća prosečna godi&scaron;nja brojnost azotofiksirajućih cijanobakterija, u povr&scaron;inskom sloju 0-5cm, je utvrđena kod zemlji&scaron;ta fluvisol u SRP &ldquo;Koviljsko petrovaradinskom ritu&rdquo;, 150864 jedinki po gramu apsolutno suvog zemlji&scaron;ta. Zemlji&scaron;te sa najnižom brojno&scaron;ću azotofiksirajućih cijanobakterija, je gajnjača u NP Fru&scaron;ka gora, 1582 jed./gr zemlji&scaron;ta u povr&scaron;inskom sloju.<br />Kod svih ispitivanih zemlji&scaron;ta brojnost azotofiksirajućih cijanobakterija je bila najveća u povr&scaron;inskom sloju zemlji&scaron;ta, 0-5 cm dubine, opadala je sa dubinom zemlji&scaron;ta i bila najmanja u najdubljem sloju, 30 - 60 cm. Kod najvećeg broja ispitivanih zemlji&scaron;ta brojnost azotofiksirajućih cijanobakterija je bila najveća tokom zimskog perioda. Azotofiksirajuće cijanobakterije su dominantne u na&scaron;im zemlji&scaron;tima i zastupljene sa 56.27% u odnosu na druge grupe algi.Izolovano je 30 sojeva azotofiksirajućih cijanobakterija: 19 sojeva Nostoc-a, 4 soja Anabaena, 4 Cylindrospermum, i po jedan soj Calothrix, Tolypothrix i Phormidium. Prosečna zastupljenost heterocista, ćelija koje vr&scaron;e azotofiksaciju, kod roda Nostoc je iznosila 8.28%, Anabaena 4.25%, Cylindrospermum-a 2.93%, Calotrix elenkinii 6.19% i Tolypothrix 7.76%.</p><p>Ultrastrukturnim ispitivanjem, TEM mikroskopom, vegetativnih ćelija azotofiksirajućih cijanobakterija uočili smo inkluzije redovnog pojavljivanja: karboksizome (Cs), cijanoficinkse granule (CG), polifosfatne granule (PG), ribozome (R), lipidne granule (&szlig; &ndash;granule) i tilakoide (T), kao i inkluzije neredovnog pojavljivanja: membranom ograničene kristalne inkluzije.<br />Koristeći TEM i tehniku bojenja ćelija sa RR i ultratankih preseka utvrdili smo prisustvo omotača od fimbrija kod tri soja (A.314, A.azollae i N.302) i tipične fimbrije kod dva soja (N.311 i N.9229). Metodom negativnog bojenja NS PTA uočili smo takođe tipične fimbrije, igličastog-dlakastog izgleda, jasnih granica niti kod tri soja (N.302, N.7901 i N.9229), međutim uočili smo i atipične sluzne fimbrije, koje nemaju jasno izražene granice, ali su veoma moćno ra&scaron;irene oko vegetativnih ćelija, kod tri soja (A.314, A.azollae, N.311).<br />Kod simbiotskih-infektivnih sojeva N.7901 i N.9229 javljaju se samo tipične fimbrije iz prve klase, a kod diazotofnih sojeva i simbiotskog - neinfektivnog soja A.azollae javljaju se atipične-sluzne fimbrije iz druge klase.<br />Za ispitivanje sličnosti cijanobakterija metodom PCR-a pomoću STRR konzervativnih sekvenci DNA genoma kori&scaron;ćeno je 39 sojeva azotofiksirajućih cijanobakterija i kod 38 je utvrđeno njihovo prisustvo. Svi sojevi se mogu podeliti u tri grupe, klastera. Prvi klaster je najveći i obuhvata 24 soja i deli se na dva podklastera: Ia koji obuhvata 12 sojeva gde dominiraju sojevi Nostoc-a (8), i podklaster Ib koji obuhvata takođe 12 sojeva, od čega 6 sojeva pripada rodu Anabaena. Podklaster Ia i podklaster Ib pokazuju različitost od 90%. Sva tri simbiozna, infektivna, soja Nostoc-a se nalaze u klasteru I: N.7901, N.9229 i N. 8001. Svaki simbiozni soj Nostoc-a ima genetske sličnosti sa po jednim diazotrofnim sojem Nostoc-a izolovanim iz zemlji&scaron;ta Vojvodine.<br />Klaster II obuhvata sedam (7) sojeva među kojima dominiraju sojevi Cylindrospermum-a, dok klaster III obuhvata 7 sojeva od čega 6 pripadaju rodu Nostoc, a jedan rodu Rivularia.Detaljno poznavanje svojstava izolovanih azotofiksirajućih cijanobakterija doprineće njihovoj budućoj primeni kako u proizvodnji ratarskih i povrtarskih kultura, tako i u biotehnolo&scaron;koj proizvodnji</p> / <p>In this study examined is the frequency of nitrogen-fixing cyanobacteria, total number of algae and total number of bacteria in different soil types on eleven localities in the Vojvodina Province. Seven out of those eleven localities are found in protected nature reserves. Actually, studied was the number of the cyanobacteria and algae depending on the depth of pedological characterization as well as on season. First, isolated were the types of nitrogen-fixing cyanobacteria, determined was their taxonomic origin and basic cytological characteristics. Also examined was the ultrastructure of vegetative cells, heterocysts and other outer structures on fimbriae and pili cells using TEM, transmission electron microscope. Finally, performed was the genetic characterization of isolated types of nitrogen-fixing cyanobacteria using the PCR method and analyzing STRR fragments of DNA.<br />The presence of nitrogen-fixing cyanobacteria and total number of algae was significantly higher with hydromorphic and halomorphic soils than with authomorphic ones. Highest annual average number of nitrogen-fixing cyanobacteria in the topsoil (0-5 cm) was reported with fluvisol soil in Special Nature Reserve &bdquo;Koviljsko petrovaradinski rit&rdquo; (Swamp) and there were 150864 units of bacteria per gram of absolutely dry soil. The soil with the lowest presence of nitrogen-fixing cyanobacteria recorded was cambisol in National Park &ldquo;Fruska gora&rdquo; with 1582 units per gram of soil in the topsoil.<br />With all the researched types of soils the number of nitrogen-fixing cyanobacteria was in the topsoil, 0-5 cm of depth and decreased in line with the depth<br />of soil and lowest was at the deepest layer, 30-60cm. The highest frequency of nitrogen-fixing cyanobacteria was found during the winter season with most of the examined soils. Nitrogen-fixing cyanobacteria are the dominant type of bacteria in our soils and are presented with 56, 27% compared to other types of algae.<br />30 strains of nitrogen-fixing cyanobacteria were isolated: 19 types of Nostoc sp., 4 of Anabaena sp. and one in each genus of Calothrix, Tolypothrix and Phormidium.<br />Using ultrastructural examination and TEM microscope when studying vegetative cells of nitrogen-fixing cyanobacteria observed were the inclusions of regular frequency: carboxysomes (Cs), cyanophycin granules (CG) , polyphosphate granules (PG), ribosomes (R), lipid granules (SS -granule ) and thylakoids ( T ) as well as the inclusion of irregular occurrence: a membrane-bound crystal inclusions.<br />Using TEM technique and staining the cells with the RR and ultra thin cross section, we determined the presence of depletion of the fimbriae with three strains (A.314, A.azollae and N.302) and typical fimbriae with the two strains (N.311 and N.9229). Applying the method of negative staining NS PTA also noticed were a typical fimbriae, needle-hairy like looks with clear boundaries with the three strains (N.302, N.7901, N.9229). However, also observed were atypical mucous fimbriae, which do not have clearly expressed borders, but they are very strongly spread around the vegetative cells, with the three strains (A.314, A.azollae, N.311).<br />With symbiotic - infective strains N.7901 and N.9229 only typical fimbriae of first class occurred, and in diazotroph strains and symbiotic &ndash; non infectious strain A. azollae found were atypical mucous fimbriae of second class.<br />To test the similarity of cyanobacteria by PCR method and using a STRR - conservative DNA sequence of the genome used were 39 strains fixing cyanobacteria and with 38 established was their presence. All strains can be divided into three groups of clusters. The first cluster is the largest and covers 24 strains, and is divided into two subclusters: Ia which includes 12 strains,where predominant are Nostoc strains ( 8 ) , and subcluster Ib , which also implies 12 strains , out of which 6 strains belong to the genus Anabaena. Subcluster Ia and Ib show a difference of 90 %. All three symbiotic , infectious Nostoc strains are classified in a cluster I: N.7901, N.9229 and N. 8001. Each symbiotic Nostoc strain has a genetic similarity with one di-nitrogen Nostoc strain isolated from a lot of different soils in Vojvodina.<br />Cluster II includes seven (7) strains , including strains among which the predominant are Cylindrospermum ones , while cluster III includes 7 strains of which 6 belong to the genus Nostoc and one to genus Rivularia.<br />Detailed knowledge of the properties of isolated fixing cyanobacteria could contribute to their future application both in the production of field crops and vegetables, as well as in biotechnological production.</p>