Submitted by Ricardo Cedraz Duque Moliterno (ricardo.moliterno@uefs.br) on 2016-09-15T22:23:03Z No. of bitstreams: 1 Tese-Maria do Socorro.pdf: 2248544 bytes, checksum: 195ed518eb9bcdd7d8346e1af11ce2bc (MD5) / Made available in DSpace on 2016-09-15T22:23:03Z (GMT). No. of bitstreams: 1 Tese-Maria do Socorro.pdf: 2248544 bytes, checksum: 195ed518eb9bcdd7d8346e1af11ce2bc (MD5) Previous issue date: 2015-10-23 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Brazil is one of the main centers of genetic diversity and dispersion of the genus Passiflora. In the present work, several species and interspecific hybrids of Passiflora were characterized through cytogenetic techniques and ISSR markers, as well as these genotypes were evaluated for in vitro multiplication and establishment. Chromosomal preparations were submitted to conventional staining, CMA3/DAPI banding, FISH using 45S/5S rDNA probes and GISH. Genomic DNA of P. edulis and P. cincinnata were used as genomic probes in the GISH. Twenty ISSR primers were used for the characterization by molecular markers. Nodal segments of P. cincinnata, P. laurifolia, P. setacea and P. luetzelburgii were collected and evaluated in the WPM, DKM and MS culture media to establishment of in vitro culture; for in vitro multiplication, explants were inoculated in DKW culture media under different concentrations of BAP, KIN and 2iP cytokinins. The results showed 2n = 18 chromosomes for the species and hybrids, regular meiosis, one pair of 5S rDNA sites and two or three pairs of 45S rDNA sites co-localized with the CMA3+/DAPI- bands. It is the first chromosomal counting recorded for P. luetzelburgii. Three groups distinct of chromosomes were revealed by GISH. The hybrid origin and the relationship with the parental species were also confirmed by using of the ISSR markers. DKW culture medium provided the better development of the explants, P. cincinnata CPE16 showed the better in vitro morphogenetic response, and KIN showed to be more efficient in obtaining increased shoots length. / O Brasil ? um dos principais centros de dispers?o da variabilidade gen?tica do g?nero Passiflora. O presente trabalho objetivou caracterizar esp?cies e h?bridos interespec?ficos de Passiflora, atrav?s de t?cnicas citogen?ticas e por marcadores ISSR, al?m de avaliar o estabelecimento e multiplica??o in vitro de gen?tipos de maracujazeiro. Prepara??es citol?gicas foram submetidas ? an?lise convencional, dupla colora??o com os fluorocromos CMA3/DAPI, FISH utilizando como sonda DNAr 5S/45S e GISH. O DNA gen?mico de P. edulis e P. cincinnata foram utilizados como sondas gen?micas na GISH. Foram utilizados 20 primers ISSR na caracteriza??o por marcadores moleculares. Para o estabelecimento do cultivo in vitro, segmentos nodais de P. cincinnata, P. laurifolia, P. setacea e P. luetzelburgii foram coletados e avaliados em meio de cultura WPM, DKW e MS, para a multiplica??o in vitro, os explantes foram inoculados em meio DKW sob diferentes concentra??es de citocininas BAP, KIN e 2iP. Os resultados mostraram 2n = 18 cromossomos para as esp?cies e h?bridos, al?m de meiose regular, um par de s?tios de DNAr 5S e dois a tr?s pares de s?tios de DNAr 45S que co-localizaram as bandas CMA3+/DAPI-, sendo o primeiro registro de contagem para P. luetzelburgii. Na GISH foi observada a forma??o de tr?s grupos cromoss?micos distintos. A origem h?brida e a rela??o com as esp?cies parentais foram tamb?m confirmadas pelo uso dos marcadores ISSR. No cultivo in vitro, o meio DKW proporcionou melhor desenvolvimento dos explantes, em rela??o aos gen?tipos, P. cincinnata CPE16 mostrou melhor resposta morfog?nica in vitro e KIN mostrou-se mais eficiente para obten??o de brota??es com maiores comprimentos.

Fascioliasis is a worldwide emerging snail-borne zoonotic trematodiasis with a great spreading capacity linked to animal and human movements, climate change, and anthropogenic modifications of freshwater environments. South America is the continent with more human endemic areas caused by Fasciola hepatica, mainly in high altitude areas of Andean regions. The Peruvian Cajamarca area presents the highest human prevalences reported, only lower than those in the Bolivian Altiplano. Sequencing of the complete rDNA ITS-2 allowed for the specific and haplotype classification of lymnaeid snails collected in seasonal field surveys along a transect including 2007–3473 m altitudes. The species Galba truncatula (one haplotype preferentially in higher altitudes) and Pseudosuccinea columella (one haplotype in an isolated population), and the non-transmitting species Lymnaea schirazensis (two haplotypes mainly in lower altitudes) were found. Climatic seasonality proved to influence G. truncatula populations in temporarily dried habitats, whereas L. schirazensis appeared to be more climatologically independent due to its extreme amphibious ecology. Along the southeastern transect from Cajamarca city, G. truncatula and L. schirazensis shared the same site in 7 localities (46.7% of the water collections studied). The detection of G. truncatula in 11 new foci (73.3%), predominantly in northern localities closer to the city, demonstrate that the Cajamarca transmission risk area is markedly wider than previously considered. Lymnaea schirazensis progressively increases its presence when moving away from the city. Results highlight the usefulness of lymnaeid surveys to assess borders of the endemic area and inner distribution of transmission foci. Similar lymnaeid surveys are still in need to be performed in the wide northern and western zones of the Cajamarca city. The coexistence of more than one lymnaeid transmitting species, together with a morphologically indistinguishable non-transmitting species and livestock movements inside the area, conform a complex scenario which poses difficulties for the needed One Health control intervention. / Ministerio de Economía y Competitividad / Revisión por pares

- ABSTRACT - Among vertebrates, reptiles represent the ideal group for the study of sex determination. Reptiles include lineages with environmental sex determination (ESD) as seen in crocodiles and tuatara, lineages with genotypic sex determination (GSD), like e.g. iguanas, chameleons, skinks, lacertid lizards and birds, and few groups which possess variability in sex determination mechanisms, i.e. geckos, dragon lizards and turtles. This thesis is focused on the evolution of sex chromosomes and sex determination in turtles. The majority of turtle species exhibit ESD, which is considered the ancestral sex determination system of this group, while GSD either as male or female heterogamety evolved independently at least five times. We investigated the presence of sex chromosomes in representative species of turtles by cytogenetic analyses. The analyses included the reconstruction of karyotypes, distribution of constitutive heterochromatin (C-banding, methylation analysis) and repetitive elements (fluorescence in situ hybridization) and comparative genome hybridization (CGH), which often characterize the degenerated Y or W and can be helpful in the identification of "cryptic" sex chromosomes. We described XX/XY sex chromosomes in seven previously unstudied Australasian chelids (Pleurodira) from the genera...

- ABSTRACT - Snakes (Serpentes) are a group of squamate reptiles (Squamata) that represents more than one third of the total reptile species diversity. Snake karyotype is generally conserved with the most common chromosome number of 36 (16 macro- and 20 microchromosomes) in diploid state. It is believed that this karyotype was also present in the common ancestor of all snakes. The majority of snake species belong to the group Caenophidia and share homologous ZW sex chromosomes. Snakes from the groups "Scolecophidia" and "Henophidia" have mostly poorly differentiated, homomorphic sex chromosomes, which made them impossible to distinguish from the autosomes in the past. These snakes were for many years assumed to have ZW sex chromosomes as well. However, recent studies demonstrated not only ZW but also two non- homologous XY sex chromosome systems in non-caenophidian snakes and thus the sex determination systems in snakes are much more variable than previously thought. In this thesis, eight species of henophidian snakes (representatives from the genera Eryx, Cylidrophis, Python and Tropidophis) and one caenophidian species (Ophiophagus hannah) were examined using conventional and molecular cytogenetic methods. However, sex chromosomes were not detected in the henophidian species, only in Ophiophagus hannah,...

Trichomonads (Parabasalia) are anaerobic microeukaryotes classified in the supergroup Excavata. Inclusion of parabasalids within Excavata is exclusively based on the molecular- phylogenetic evidence. Over 400 species of parabasalids have been described so far, and the vast majority of them are endobiotic. In contrast, only few species of free-living parabasalids forming four independent lineages have been described (Pseudotrichomonas keilini, Ditrichomonas honigbergii, Monotrichomonas carabina, Honigbergiella sp., Tetratrichomonas undula, and Lacusteria cypriaca). Lacusteria cypriaca is a new species and genus described in our recent paper. In this paper we published the first two sequences of SSU rDNA from Pseudotrichomonas keilini as well. All of these lineages are likely secondarily free-living, and they developed from endobiotic ancestors. In addition to the already published Lacusteria cypriaca and Pseudotrichomonas keilini strains, we have recently obtained seven another isolates of free-living trichomonads (LAGOS2D, E2NT, CK, LAGOS2M, GR8, GOU23 LIVADIAN, and VAV1A1); from all of these isolates we sequenced SSU rDNA and performed phylogenetic analyses. These isolates split into four independent evolutionary lineages, which indicate that free-living parabasalids are more diversed and...

Actualmente, en el tratamiento de aguas residuales no se presta demasiada atención a los microorganismos que realizan la depuración ni interfieren en el proceso. Es por ello que con este estudio se pretende estudiar este tipo de microorganismos y conocer qué diversidad de microorganismos se puede encontrar, así como encontrar aplicaciones biotecnológicas a partir de ellos. El objetivo principal del trabajo es, mediante la taxonomía polifásica, estudiar la biodiversidad de actinomicetos productores de espumas, además de estudiar su capacidad de biodegradación de ciertos productos tóxicos. Esto se realizará mediante el aislamiento de la bacteria, su caracterización quimiotaxonómica (incluyendo extracción de ácidos micólicos, determinación de los isómeros del ácido diaminopimélico y la extracción de los azúcares predominantes e la pared celular), caracterización genotípica (que incluye extracción de DNA, amplificación por PCR, secuenciación y construcción e interpretación de árboles filogenéticos), caracterización fenotípica (realización de diferentes tests fenotípicos), estudios de biodegradación de productos tóxicos derivados del petróleo (fenol y naftaleno) y detección molecular del gen catecol 1,2-dioxigenasa para determinar qué aislados poseen el potencial catabólico para degradar hidrocarburos aromáticos. Con todo ello, se obtuvieron 152 aislados, pertenecientes a 28 estaciones depuradoras de aguas residuales, pertenecientes al suborden phylum Actinobacteria. Tras realizar la caracterización quimiotaxonómica se obtuvo que 147 de ellos pertenecían al suborden Corynebacterineae, 4 al suborden Pseudonocardiaceae y 1 al suborden Micrococcineae. Dentro de estos subórdenes, los aislados pertenecían a 9 géneros diferentes y a 37 especies diferentes, lo que mostró la gran diversidad existente en estos ambientes. Los ensayos de biodegradación mostraron que 76 aislados eran capaces de degradar al menos un producto tóxico (fenol o naftaleno). / Soler Hernández, A. (2012). Biodiversidad de actinomicetos aislados de plantas depuradoras de aguas residuales. Estudio de la capacidad de biodegradación de compuestos tóxicos [Tesis doctoral]. Editorial Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/14982 / Palancia

No description available.

No description available.

Rhizoctonia blight (sensu lato) is a common and serious disease of many turfgrass species. The most widespread causal agent R. solani consists of several genetically different anastomosis groups (AGs) and subgroups. Though anastomosis or hyphal fusion reactions have been used to group Rhizoctonia species, they are time consuming and sometimes difficult to interpret. Anastomosis reactions are incapable of identifying isolates belonging to different AG subgroups within an AG. This study evaluated molecular techniques in comparison with traditional anastomosis grouping (AG) to identify and group isolates of Rhizoctonia. More than 400 Rhizoctonia isolates were collected from diseased turfgrass leaves from eight geographic areas in Virginia and Maryland. A random sample of 86 isolates was selected and initially characterized by colony morphology, nuclei staining and anastomosis grouping. Molecular identification was performed by analysis of rDNA-ITS region and DNA fingerprinting techniques universally primed PCR (UP-PCR) and amplified fragment length polymorphism (AFLP). The cladistic analysis of ITS sequences and UP-PCR fragments supported seven clusters. Isolates of R. solani AG 1-IB (n=18), AG 2-2IIIB (n=30) and AG 5 (n=1) clustered separately. Waitea circinata var. zeae (n=11), and var. circinata (n=4) grouped separately. A cluster of six isolates (UWC) did not fall into any known Waitea group. Most of the binucleate Rhizoctonia-like fungi (BNR) (n=16) grouped separately. AFLP grouping also largely agreed with the above results. However, UWC isolates clustered into two groups. Molecular analyses corresponded well with traditional anastomosis grouping by clustering isolates within an AG or AG subgroup together. UP-PCR cross-hybridization could distinguish closely related Rhizoctonia isolates to their infraspecies level. Genetically related isolates belonging to the same AG subgroups cross-hybridized strongly, while isolates of different AGs did not cross-hybridize or did so weakly. Sequence-characterized amplified region (SCAR) markers were generated from UP-PCR products to identify isolates of major pathogenic groups AG 1-IB and AG 2-2IIIB. Specific primer pairs successfully distinguished isolates of AG 1-IB and AG 2-2IIIB from isolates of other AGs. Sensitivity of Rhizoctonia species and AGs was tested in vitro to commercial formulations of iprodione, triticonazole and pyraclostrobin. W. circinata isolates were moderately sensitive to iprodione while isolates of R. solani and BNR were extremely sensitive. Isolates of AG 2-2IIIB showed less sensitivity to triticonazole than other Rhizoctonia isolates. W. circinata var. zeae isolates were moderately sensitive to pyraclostrobin while most of the other isolates were extremely sensitive. / Ph. D.

Salmonella enterica serotypes have been linked to outbreaks associated with low water activity foods. The ability of biofilm forming pathogens, such as Salmonella, to survive thermal and chemical processes is improved; it is unclear if biofilms will also improve survival to desiccation and gastric stresses. The purpose of this study was to quantify the effect of physiological state (planktonic versus biofilm) and prior exposure to desiccation on Salmonella survival and gene expression after passage through an in-vitro digestion model. Cells of Salmonella enterica serotype Tennessee were deposited onto membranes for planktonic cells or on glass beads to create biofilms. The cells were subsequently dried at room temperature and stored in dried milk powder (aw = 0.3) for up to 30 days. Salmonella survival was quantified by serial dilution onto brilliant green agar before desiccation, after desiccation, after 1-day storage and after 30-day storage. At each sampling both physiological states were tested for survival through a simulated gastrointestinal system. RNA was extracted at the identical time points and relative gene expression determined for genes associated with stress response (rpoS, otsB), virulence (hilA, hilD, invA, sipC) and a housekeeping gene 16S rRNA using quantitative real-time PCR. The physiological state and length of storage effected the survival and gene expression of Salmonella within the desiccated milk powder environment and after passage through an in-vitro digestion system (p<0.05). Larger numbers of S. Tennessee were recovered by plate counts for biofilm cells, compared to planktonic cells. However, the numbers of 16S rRNA gene copies were not significantly different suggesting entry of S. Tennessee into a viable but non-culturable state. Prolonged storage in dry milk powder was not associated with increased cross-protection to gastric stress. Increased expression of stress response genes rpoS and otsB correlated with survival, indicating cross protection of low water activity and acid stress. Increased expression of virulence-associated genes was seen in cells exposed to short periods of dry storage, suggesting an increased virulence potential. / Master of Science in Life Sciences