This study applies molecular and morphological procedures to identify larval and adult stages of trematodes of the family Strigeidae in Iceland. Intermediate hosts (snails and fishes) and definitive host (birds) from 11 freshwater lakes were sampled and examined for the presence of trematodes. Recovered species were subjected to study of morphology, sequencing and phylogenetic analyses. A total of seven species of three genera were identified via phylogenetical analyses based on mitochondrial (cox1) and nuclear (28S) sequences, and morphological data. The life-cycle of Apatemon gracilis was fully elucidated in Iceland, and those of Australapatemon burti and Australapatemon minor in part (cercariae and adults). The relationship of Cotylurus sp. 'Lin. 1I' and Cotylurus sp. 'Lin. 2I' could not be resolved.

The Crepidotaceae (Imai) Singer (Basidiomycetes: Agaricales) represents a proposed family of saprophytic fungi containing five agaricoid (Crepidotus, Tubaria, Melanomphalia, Simocybe, Pleurotellus) and four cyphelloid (Episphaeria, Phaeosolenia, Pellidiscus, Chromocyphella) genera. Several contemporary classification systems exist that delegate some or all of these genera to other agaric families. Phylogenetic relationships for the most prevalent genera in the Crepidotaceae were investigated using nuclear large subunit ribosomal DNA (LSU rDNA) sequences. Parsimony analysis of the molecular data supports the Singer classification of Crepidotus, Melanomphalia, and Simocybe as a single monophyletic unit within the Agaricales. The affinities of the genus Tubaria remain uncertain. Crepidotus (Fr.) Staude is the largest and most phenotypically variable genus in the Crepidotaceae. Sequencing of the LSU rDNA region for a cross-section of morphologically diverse species suggests that Crepidotus is not a monophyletic genus. Analysis of morphological characters for 23 Crepidotus taxa shows that characters traditionally applied for infrageneric classification of Crepidotus are homoplasic in origin, but that less commonly emphasized characters such as spore shape and ultrastructure of spore wall ornamentation may be indicative of monophyletic clades for this complex. A unique pattern of basidiospore dormancy and germination, unknown in any other species of agaric, is reported for 11 species of Crepidotus. Similar patterns were also encountered in species of Simocybe and Melanomphalia. In these species an endogenous period of spore dormancy of four to six months is followed by an activation period where the factors necessary for subsequent germination appear to involve a minimal nutritional component, water, and exposure to light. / Master of Science

BACKGROUND:Numerous studies have investigated cospeciation between parasites and their hosts, but there have been few studies concerning parasitoids and insect hosts. The high diversity and host specialization observed in Anicetus species suggest that speciation and adaptive radiation might take place with species diversification in scale insect hosts. Here we examined the evolutionary history of the association between Anicetus species and their scale insect hosts via distance-based and tree-based methods.RESULTS:A total of 94 Anicetus individuals (nine parasitoid species) and 113 scale insect individuals (seven host species) from 14 provinces in China were collected in the present study. DNA sequence data from a mitochondrial gene (COI) and a nuclear ribosomal gene (28S D2 region) were used to reconstruct the phylogenies of Anicetus species and their hosts. The distance-based analysis showed a significant fit between Anicetus species and their hosts, but tree-based analyses suggested that this significant signal could be observed only when the cost of host-switching was high, indicating the presence of parasite sorting on related host species.CONCLUSIONS:This study, based on extensive rearing of parasitoids and species identification, provides strong evidence for a prevalence of sorting events and high host specificity in the genus Anicetus, offering insights into the diversification process of Anicetus species parasitizing scale insects.

SANTRAUKA PAUKŠČIUOSE APTINKAMŲ NEAPIBŪDINTŲ SARCOCYSTIS GENTIES RŪŠIŲ FILOGENETINIŲ RYŠIŲ TYRIMAS NAUDOJANT DALINES 18S IR 28S rRNR GENŲ SEKAS Petras Prakas Vilniaus Universiteto Ekologijos Institutas, Populiacinės Genetikos Laboratorija, Vilnius, Lietuva. Pagal cistų morfologiją, baltakaktėje žąsyje (Anser albifrons) aptiktos I tipo sarkocistos, didžiojoje antyje (Anas platyrhynchos) rastos II tipo sarkocistos, dviejose baltakaktėse žąsyse ir pilkojoje žąsyje (Anser anser) nustatytos III tipo sarkocistos, varnoje (Corvus cornix) identifikuotos V tipo sarkocistos. Iš paukščių, kaip tarpinių šeimininkų išskirtos sarkosporidijos aprašomos pirmą kartą bei yra neįvardintos rūšys. Baltakaktėje bei pilkojoje žąsyse išskirtos III tipo sarkocistos gali būti tos pačios sarkocistų rūšies parazitavimo skirtingų rūšių tarpiniuose šeimininkuose atvejis. Nustatytos 18S ir 28S rDNR fragmentų sekos pasižymėjo didžiausia homologija Sarcocystis bei Frenkelia gentims. Filogenetinių ryšių medyje sugrupuojamos šiame darbe tiriamos rūšys su Frenkelia microti, Frenkelia glareoli, Sarcocystis muris, Sarcocystis neurona. Rezultatai atskleidė Sarcocystis genties rūšių koevoliuciją su jų galutiniais šeimininkais. Remiantis tyrimo duomenimis, galima teigti, kad Sarcocystidae šeimos filogenetinius ryšius apsprendžia sarkosporidijų gyvybinio ciklo ypatybės, šeimininkų spektras bei parazitų specifiškumas šeimininkui. 18S ir 28S rRNR genų dalinių sekų analizė atskleidžia tirtų Sarcocystis sp. rūšių... [toliau žr. visą tekstą] / SUMMARY INVESTIGATION OF THE PHYLOGENETIC RELATIONSHIPS OF SARCOCYSTIS SPP. FOUND IN BIRDS USING 18S AND 28S rRNA GENE PARTIAL SEQENCES Petras Prakas Institute of Ecology of Vilnius University, Laboratory of Population Genetics, Vilnius, Lithuania Based on cyst morphology, Sarcocystis cysts type I were found in white-fronted goose (Anser albifrons), cysts type II in mallard (Anas platyrhynchos), cyst type III in one grey-lag goose (Anser anser) and two white-fronted geese, cyst type V in hooded crow (Corvus cornix). The sarcocysts isolated from the infected birds as intermediate host have not been previously described and are unnamed. Type III sarcocysts detected in white-fronted and greylag geese may illustrate the case of polyhostal nature of sarcocysts when the same-species parasites infesting intermediate hosts of different species. The obtained 18S and 28S ribosomal RNA partial gene sequences showed the highest homology for the genera Sarcocystis and Frenkelia. In the tree of phylogenetic relationships, the species involved in this study were grouped with Frenkelia microti, Frenkelia glareoli, Sarcocystis muris and Sarcocystis neurona. Results show co-evolution of Sarcocystis spp. with the final host. Based on data obtained it could be stated that phylogenetic relationships of Sarcocystidae family are influenced by peculiarity of life cycle, hosts spectrum and host specificity. Analysis of the partial sequences of 18S and 28S ribosomal RNA revealed the phylogenetic and... [to full text]

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Coccidiodomycosis is a systemic infection, predominantly pulmonary, caused by the geophilic and dimorphic fungi, Coccidioides immitis and Coccidioides posadasii. In Brazil, coccidioidomycosis is associated with semi-arid areas in the Northeastern region of this country, which is considered one of the endemic areas of this disease in South America. These pathogens are morphologically indistinguishable species, but they exhibit molecular differences. Different molecular techniques have been described for the characterization of these species. The nuclear ribosomal DNA (rDNA) from Coccidioides spp. has been described as an important molecular marker for the identification, taxonomy and phylogeny. Currently, there are still shortages of maps for epidemiological approaches in order to direct the correlation between populations of Coccidioides spp. and the outbreaks of coccidiomycosis. Given the above, this study aimed at performing the molecular identification of 18 clinical and environmental isolates of C. posadasii, from Northeastern Brazil, maintained in the fungal collection of the Specialized Medical Mycology Center (CEMM), through PCR, as well as, to analyze the genetic diversity of these isolates by sequencing of 18S-28S regions of nuclear rDNA. The identification of the isolates was performed through PCR, using specific primers Coi9-1F and Coi9-R. The sequencing of the 18S-28S rDNA regions was performed through the method of chain termination by dideoxynucleotides, using the kit DYEnamicTM ET terminators cycle sequencing (GE Healthcare). The results confirmed the identification of all strains included in this study as belonging to the species C. posadasii. The phylogenetic tree based on 18S-28S rDNA region of C. posadasii from CEMM and Coccidioides spp. from Genbank. reveals the formation of a unique cluster encompassing the following strains CEMM 05-2-063, CEMM 05-2-064, CEMM 05-2-066 and CEMM 05-2-065, in a properly sustained branch, which apparently seems to group these isolates according to their geographical origin. The strains of C. posadasii showed lower genetic divergence in the ITS1 and ITS2 regions, when compared to strains of C. immitis. Analyses did not detect differences between strains of clinical origin and those of environmental origin. Further studies involving the analysis of fast evolving markers, such as microsatellites, can provide evidences to determine whether the groups found in this study are derived from a lineage of clonal reproduction. / A coccidioidomicose à uma infecÃÃo sistÃmica, predominantemente pulmonar, causada pelos fungos dimÃrficos e geofÃlicos, Coccidioides immitis e Coccidioides posadasii. No Brasil, a coccidioidomicose està associada a locais situados na zona semi-Ãrida da regiÃo Nordeste, considerada uma das Ãreas endÃmicas da doenÃa na AmÃrica do Sul. Estes patÃgenos consistem em espÃcies morfologicamente indistinguÃveis, mas que exibem diferenÃas moleculares peculiares. O DNA ribossÃmico nuclear (rDNA) de Coccidioides spp. tem sido apontado como importante marcador molecular utilizado na identificaÃÃo, taxonomia e filogenia. Atualmente, ainda hà escassez de mapas de abordagens epidemiolÃgicas para direcionar a correlaÃÃo entre as populaÃÃes de Coccidioides spp. com os surtos de coccidioidomicose. Diante do exposto, este estudo teve por objetivo, realizar a identificaÃÃo molecular de 18 isolados clÃnicos e ambientais de C. posadasii, oriundos do Nordeste brasileiro, mantidos na Micoteca do Centro Especializado em Micologia MÃdica (CEMM), atravÃs da tÃcnica de PCR, bem como, analisar a diversidade genÃtica destes isolados por meio do seqÃenciamento das regiÃes 18S-28S do rDNA nuclear. A identificaÃÃo dos isolados foi realizada por PCR utilizando os primers especÃficos Coi9-1F e Coi9-R. O sequenciamento das regiÃes 18S-28S rDNA foi realizado pelo mÃtodo da terminaÃÃo da cadeia pelo didesoxinucleotÃdeo, usando-se o kit DYEnamicTM ET terminators cycle sequencing (GE Healthcare). Os resultados confirmaram a identificaÃÃo de todas as cepas incluÃdas neste estudo, como pertencentes à espÃcie C. posadasii. A Ãrvore filogenÃtica, baseada na regiÃo 18S-28S rDNA de C. posadasii do CEMM, juntamente com sequÃncias de Coccidioides spp. depositadas no Genbank. revela a formaÃÃo de um cluster exclusivo englobando as cepas CEMM 05-2-063, CEMM 05-2-064, CEMM 05-2-065 e CEMM 05-2-066, em um ramo adequadamente sustentado, que aparentemente parece agrupar estes isolados segundo sua origem geogrÃfica. As cepas de C. posadasii apresentaram menor Ãndice de divergÃncia genÃtica nas regiÃes ITS1 e ITS2, quando comparadas Ãs cepas de C. immitis A anÃlise nÃo detectou diferenÃas entre as cepas de origem clÃnica e as de origem ambiental. Estudos posteriores envolvendo a anÃlise de marcadores de evoluÃÃo mais rÃpida, os microssatÃlites, podem fornecer evidÃncias para determinar se os agrupamentos encontrados neste estudo sÃo resultantes de uma linhagem de reproduÃÃo clonal.

Parasitism is a common phenomenon in nature, where one species benefits at the expense of another. In marine environments, fungal parasites are known to infect phytoplankton, causing disease and death, and contributing to the loss of phytoplankton populations. The impact of fungal parasites on phytoplankton communities can be significant, as selective parasitism on one species of phytoplankton can encourage the development of other species and may be one of the factors influencing communities' seasonal succession. Fungi parasites play a crucial role in sinking by inducing cell death and promoting cell aggregation, which can lead to the export of phytoplankton to the deep sea. Although fungal parasites are well studied in freshwater phytoplankton, there is still little information on marine phytoplankton parasites. Recent studies have shown that large diatoms are the preferred hosts for fungal parasites in cold waters, while the infection of dinoflagellates is reported in the oligotrophic Mediterranean Sea. However, the occurrence and impact of fungal parasites on phytoplankton communities in the Red Sea remain largely unknown. Therefore, the proposed project aims to investigate the occurrence of fungal parasites that infect phytoplankton and analyze their role in the export of phytoplankton to the deep sea in the Red Sea. By understanding the ecological impacts, life cycle, host range, and infection strategies of fungal parasites on populations of Red Sea phytoplankton, this study can provide critical information on the dynamics of marine ecosystems and carbon cycling.

Submitted by Angela Maria de Oliveira (amolivei@uepg.br) on 2017-10-19T19:19:13Z No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) Rafael Bonfim de Almeida.pdf: 1398729 bytes, checksum: b49f522af530568d079f7e0bdc9a7372 (MD5) / Made available in DSpace on 2017-10-19T19:19:13Z (GMT). No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) Rafael Bonfim de Almeida.pdf: 1398729 bytes, checksum: b49f522af530568d079f7e0bdc9a7372 (MD5) Previous issue date: 2013-02-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A grande quantidade de dados filogenéticos que vêm sendo disponibilizados, principalmente de caráter molecular, com os avanços de técnicas de obtenção, amplificação e sequenciamento de moléculas de DNA, RNA e proteínas, oferece uma nova perspectiva para a resolução de relações filogenéticas de grupos problemáticos. Porém, esta crescente quantidade de dados necessita de estratégia de integração e análise conjunta tanto entre os dados moleculares a per se quanto com outros dados anteriores, como dados morfológicos, citogenéticos, etológicos, entre outros. Uma alternativa recente para essa integração e metanálise filogenética é a abordagem de construção de super árvores, que consiste no princípio de agrupamento de filogenias independentes menores que possuem táxons terminais em comum em uma filogenia maior, construída com maior número de caracteres, com maior número de táxons terminais, sendo, portanto uma importante ferramenta para inferências mais acuradas quanto à sistemática, evolução, biologia, etologia, biogeografia e citogenética já que oferece uma perspectiva mais ampla do grupo em estudo. Neste estudo, dois grupos problemáticos quanto à sistemática filogenética foram escolhidos e analisados sob a perspectiva da abordagem de construção de super árvore. Para a subfamília Galerucinae sensu lato foi realizada uma filogenia molecular para espécies de Alticinae com ênfase na tribo Oedionychini a partir do alinhamento das sequências do segmento de expansão 2 (D2) do gene nuclear para 28S rDNA (28S-D2) e o gene mitocondrial para a subunidade I do Citocromo Oxidase C (COI), sendo essa filogenia juncionada com sequências associadas a bancos de dados e utilizadas em estudos filogenéticos anteriores. Posteriormente essa filogenia molecular foi integrada a uma super árvore construída para Galerucinae sensu lato. A super árvore recuperou a monofilia de Galerucinae com um grupo Alticinae parafilético. Para a ordem Siluriformes (Teleostei: Ostariophysi) foi construída uma super árvore baseada tanto em dados morfológicos quanto dados moleculares, contendo todas as famílias do grupo na topologia final. A topologia final recuperou a monofilia de Siluriformes, com Diplomystidae sendo o silurídeo mais basal e grupo-irmão do restante dos Eusiluroidei. / The large amount of phylogenetic data that are being available, with predominance of molecular, with the recent advances in techniques to obtainment, amplification and sequencing of DNA molecules, RNA and proteins, offer a new perspective for solving phylogenetic relationships of problem groups. However, this increased amount of data requires integration strategies and combined analysis of data, gathering the molecular per se with other previous data, such as morphological, cytological, ethological, and others. A recent alternative for this integration and phylogenetic meta-analysis is the Supertree Construction approach, that in principle consists in a gathering of independent smaller phylogenies that have some terminal taxa in common into a larger phylogeny, built with more characters, major number of terminal taxa, being an important tool for more accurate inferences about the systematics, evolution, biology, ethology, biogeography, cytogenetics, and others, once it offers a broader perspective of the study group. In this study, two problematic groups about their phylogenetic systematics were selected and analyzed under the perspective of the Supertree Construction approach. For subfamily Galerucinae sensu lato was performed a molecular phylogeny for Alticinae species with emphasis on the segment Oedionychini, based on alignment of the sequences of expansion segment 2 (D2) of nuclear 28S rDNA gene (28SD2), and the mitochondrial gene for the subunit I of cytochrome oxidase c (COI), being this phylogenies gather with other data used in previous phylogenetic studies. Later, this molecular phylogeny was incorporated into a supertree, constructed for Galerucinae sensu lato. The supertree recovered the monophyly of a Galerucinae group, and paraphyletic Alticinae. To order Siluriformes (Teleostei: Ostariophysi) was built a supertree based on both, morphological and molecular data, containing all families recognized for the group. The final topology recovered the monophyly of Siluriformes with Diplomystidae being the most basal catfish and sister group of the rest of Eusiluroidei.

Made available in DSpace on 2014-12-17T14:10:26Z (GMT). No. of bitstreams: 1 TiaraSC_DISSERT_PARCIAL.pdf: 3904486 bytes, checksum: c7dbd69e3fa57b6310c7eaf6781c6f83 (MD5) Previous issue date: 2011-07-15 / Gasteroids fungi are characterized by the basidiospores maturation inside the basidioma, from which spores liberation occurs in a passive manner. These fungi were once seen as a well definite class of Basidiomycota, but nowadays they are considered an artificial assemblage, because the organisms have independent evolutionary histories forming a polyphyletic group with a vast morphological variety. Despite their diversity, studies with this group in the tropics are incipient, and the phylogenetic relationships of the species from temperate climate remain unknown. Thus, this work aimed to elucidate the phylogenetic relationships of gasteroids fungi from the Geastrales and Phallales orders, with the inclusion of tropical and temperate species, and with these analyses suggest a systematic position of species like Asero? floriformis and Phallus roseus, as well as to verify if the lignicolous habit can indicate parental relationship in the Geastrum genus. For this, basidiomata were collected at Atlantic rain forest areas, during the rainy season, and the specimen identification followed specific literature for gasteroid fungi. The phylogenetic analyses were performed with Maximum Parsimony and Bayesian Analysis, making use of RPB2 and 28S nuclear genes and atp6 mitochondrial gene. It could be observed on the Phallales dendogram, that Asero? floriformis did not cluster with A. rubra, and that it has an anterior divergence from all others species of the family Clathraceae used in this analysis, assuming a basal position in the clade. Phallus roseus, which once was recognized as Itajahya, has previous divergence from the group formed by Phallus species. At the Geastrales dendogram, in the group corresponding to Geastrum genus, it could be observed that species with lignicolous habitat clustered in a clade with high support values. So, the results suggest the creation of a new genus to accommodate A. floriformis, and the revalidation of Itajahya, as well as it can be affirmed that the lignicolous habitat on the Geastrum genus in fact indicates parental relationships, and that it has arised only once at the evolutionary history of the genus / Os fungos gasteroides s?o reconhecidos pela matura??o dos basidi?sporos dentro do basidioma, cuja libera??o ocorre de forma passiva. Esses fungos j? foram vistos como uma classe bem definida de Basidiomycota, mas atualmente s?o considerados um grupo artificial, por tratar-se de organismos com hist?rias evolutivas independentes formando um grupo polifil?tico com grande diversidade morfol?gica. Apesar da grande diversidade, estudos com esse grupo nos tr?picos ainda ? incipiente, e as rela??es filogen?ticas com esp?cies de regi?es temperadas permanecem desconhecidas. Dessa forma, objetivou-se neste trabalho elucidar as rela??es filogen?ticas de fungos gasteroides das ordens Geastrales e Phallales, com a inclus?o de esp?cies tropicais e de regi?es temperadas, e atrav?s dessas an?lises conhecer a rela??o de Asero? floriformis e Phallus roseus com outros membros do grupo, assim como verificar se o h?bito lign?cola em Geastrum pode indicar rela??o de parentesco. Para isso, coletas de basidiomas foram realizadas em ?reas de Mata Atl?ntica em per?odo chuvoso, com a identifica??o dos esp?cimes seguindo bibliografia espec?fica para fungos gasteroides. Para as an?lises filogen?ticas foram utilizados os m?todos de M?xima Parcim?nia e An?lise Bayesiana, utilizando-se os genes nucleares RPB2 e 28S, e o gene mitocondrial atp6. Observou-se no dendograma obtido para a ordem Phallales, que Asero? floriformis n?o se agrupou com A. rubra, apresentando diverg?ncia anterior a todas as esp?cies da fam?lia Clathraceae utilizadas na an?lise, assumindo uma posi??o basal no clado. J? Phallus roseus, antes reconhecido como Itajahya, possui diverg?ncia anterior ao grupo formado pelas esp?cies de Phallus. No dendograma da ordem Geastrales, no grupamento correspondente ao g?nero Geastrum, p?de-se observar esp?cies que possuem h?bito lign?cola se agrupando com alto valor de suporte. Assim, os resultados sugerem a cria??o de um novo g?nero para acomodar A. floriformis, e a revalida??o do g?nero Itajahya, assim como se pode afirmar que o h?bito lign?cola em Geastrum de fato indica rela??es de parentesco, aparentemente tendo surgindo apenas uma vez na hist?ria evolutiva do grupo

<p>Род <em>Merodon Meigen</em> (Diptera: Syrphidae) се одликује великм бројем врста које имају&nbsp; функцију у опрашивању&nbsp; биљака. У оквиру овог рода својом разноврсношћу издваја се <em>Merodon aureus</em>&nbsp; група&nbsp; врста коју поред фенотипски различитих одликује и присуство већег броја криптичних&nbsp; врста. Због морфолошке сличности криптичне врсте представљају изазов за таксономе, те су молекуларне методе од посебног значаја. У том светлу примарни циљ овог истраживања је био утврђивање молекуларног диверзитета групе и могућности његове примене у таксономији. Истраживање је било засновано на анализи варијабилности секвенци&nbsp; COI&nbsp; и&nbsp; 28S&nbsp; рРНК гена за 718 јединки 41 описане или новоткривене врсте сирфида. Додатно, тестирана је примењивост мултилокусних&nbsp; ISSR маркера у раздвајању врста&nbsp; <em>M.&nbsp; luteomaculatus</em>&nbsp; комплекса. Узорци су тестирани и на присуство<em> Wolbachia</em> с обзиром да она може утицати на еволуцију митохондријалних гена. Добијени резултати су указали на високу варијабилност секвенци&nbsp; COI&nbsp; гена&nbsp; који се показао корисним у&nbsp; утврђивању граница&nbsp; криптичних&nbsp; врста у анализираним комплексима. Секвенце 28Ѕ рРНК гена у већини случајева нису биле од већег значаја за раздвајање врста, али би могле имати потенцијал у&nbsp; раздвајању комплекса&nbsp; или подгрупа&nbsp; врста као допуна анализи&nbsp; секвенци&nbsp; COI&nbsp; гена. Додатно,&nbsp; ISSR&nbsp; маркери су показали потенцијал за примену у молекуларној таксономији. Све анализиране врсте изузев <em>М. balkanicus</em> су биле заражене&nbsp; <em>Wolbachia,</em> али није утврђена јасна веза између варијабилности&nbsp; секвенци&nbsp; митохондријалног&nbsp; COI гена и инфекционог статуса врста. Утврђен образац генетичке варијабилности је вероватно&nbsp; обликован драстичним климатским променама&nbsp; током Плеистоцена и фрагментацијом хабитата. Да би се донели крајњи закључци о таксономском статусу предложених врста потребно је добијене резултате интегрисати са подацима других релевантних&nbsp; научних дисциплина.</p> / <p>Rod <em>Merodon Meigen</em> (Diptera: Syrphidae) se odlikuje velikm brojem vrsta koje imaju&nbsp; funkciju u oprašivanju&nbsp; biljaka. U okviru ovog roda svojom raznovrsnošću izdvaja se <em>Merodon aureus</em>&nbsp; grupa&nbsp; vrsta koju pored fenotipski različitih odlikuje i prisustvo većeg broja kriptičnih&nbsp; vrsta. Zbog morfološke sličnosti kriptične vrste predstavljaju izazov za taksonome, te su molekularne metode od posebnog značaja. U tom svetlu primarni cilj ovog istraživanja je bio utvrđivanje molekularnog diverziteta grupe i mogućnosti njegove primene u taksonomiji. Istraživanje je bilo zasnovano na analizi varijabilnosti sekvenci&nbsp; COI&nbsp; i&nbsp; 28S&nbsp; rRNK gena za 718 jedinki 41 opisane ili novotkrivene vrste sirfida. Dodatno, testirana je primenjivost multilokusnih&nbsp; ISSR markera u razdvajanju vrsta&nbsp; <em>M.&nbsp; luteomaculatus</em>&nbsp; kompleksa. Uzorci su testirani i na prisustvo<em> Wolbachia</em> s obzirom da ona može uticati na evoluciju mitohondrijalnih gena. Dobijeni rezultati su ukazali na visoku varijabilnost sekvenci&nbsp; COI&nbsp; gena&nbsp; koji se pokazao korisnim u&nbsp; utvrđivanju granica&nbsp; kriptičnih&nbsp; vrsta u analiziranim kompleksima. Sekvence 28Ѕ rRNK gena u većini slučajeva nisu bile od većeg značaja za razdvajanje vrsta, ali bi mogle imati potencijal u&nbsp; razdvajanju kompleksa&nbsp; ili podgrupa&nbsp; vrsta kao dopuna analizi&nbsp; sekvenci&nbsp; COI&nbsp; gena. Dodatno,&nbsp; ISSR&nbsp; markeri su pokazali potencijal za primenu u molekularnoj taksonomiji. Sve analizirane vrste izuzev <em>M. balkanicus</em> su bile zaražene&nbsp; <em>Wolbachia,</em> ali nije utvrđena jasna veza između varijabilnosti&nbsp; sekvenci&nbsp; mitohondrijalnog&nbsp; COI gena i infekcionog statusa vrsta. Utvrđen obrazac genetičke varijabilnosti je verovatno&nbsp; oblikovan drastičnim klimatskim promenama&nbsp; tokom Pleistocena i fragmentacijom habitata. Da bi se doneli krajnji zaključci o taksonomskom statusu predloženih vrsta potrebno je dobijene rezultate integrisati sa podacima drugih relevantnih&nbsp; naučnih disciplina.</p> / <p>Genus<em>&nbsp; Merodon&nbsp; Meigen&nbsp;</em> (Diptera: Syrphidae)&nbsp; is characterized by a large number of species which have a function in plants pollination. Within this genus, <em>Merodon </em>aureus species group is distinguished by its diversity and it is characterized not only by phenotypically divergent species but also with a large number of cryptic species. Because of the high morphological similarity, cryptic species are a challenge for taxonomist, thus molecular methods are of special importance. The primary goal of this research was to determine the molecular diversity of the group and the possibility of its application in taxonomy. The study was based on the sequences variability analysis of COI and 28S rRNA genes for 718 individuals belonging to 41 described or newly discovered hoverfly species. Additionally, the applicability in species delimitation of multilocus marker ISSR was tested on&nbsp; <em>M. luteomaculatus</em>&nbsp; species complex. Specimens have also been tested for<em> Wolbachia</em> since&nbsp; it may affect the evolution of mitochondrial genes. The obtained results indicated a high variability of the COI gene&nbsp; sequences that proved useful for determining the&nbsp; cryptic&nbsp; species boundaries in the analyzed complexes. Sequences of the 28S rRNA gene in&nbsp;&nbsp; most cases were not of much significance for the species delimitation, but they could have the potential to separate species complexes&nbsp; оr subgroups&nbsp; as a complement to the analysis of the COI gene&nbsp; sequences. Additionally, the ISSR markers showed potential for application in molecular taxonomy. All analyzed species&nbsp; except&nbsp; <em>M. balkanicus</em>&nbsp; were infected with&nbsp; <em>Wolbachia</em>, but no clear&nbsp; relation&nbsp; was established between the sequence variability of the mitochondrial COI gene and&nbsp; the infectious status of species. The established pattern of genetic variability is probably shaped&nbsp; by drastic climatic changes during Pleistocene and habitats fragmentation. In order to&nbsp; achieve&nbsp; the final conclusions on the taxonomic status of the proposed species, it is necessary to integrate the obtained results with the data of other&nbsp; relevant scientific disciplines.</p>

Chaque année, environ 175 millions de personnes souffrent d’infection du tractus urinaire (ITU) dans le monde. Pour diagnostiquer une ITU, une culture d’urine est réalisée mais nécessite un long délai. Pendant ce temps les patients sont souvent traités avec un antibiotique à large spectre qui peut entrainer des complications et contribuer à l’augmentation du nombre de bactéries résistantes. La plupart des échantillons reçus au laboratoire sont négatifs (≤10[indice supérieur 5] ufc /mL) ou contiennent Escherichia coli. Nous avons mis au point une méthode de dépistage rapide des échantillons positifs afin de sauver des coûts et éviter des complications. D’abord un protocole d’extraction d’ADNg a été élaboré avec la même efficacité pour tous les microorganismes. Cette méthode, appelé EtNa, chauffe le microorganisme dans une solution de 70 % éthanol et NaOH. Cette solution permet l’adsorption de l’ADN directement sur une colonne de silice afin de favoriser sa purification par un robot pipeteur. Elle a été comparée favorablement avec d’autres méthodes retrouvées dans la littérature et trousses commerciales, mais a l’avantage d’extraire les bactéries Gram positifs, Gram négatifs et levures avec une efficacité similaire et ce, avec un même protocole simple et sans produits chimiques toxiques. La deuxième étape du projet a été de mettre au point une méthode pour dépister les microorganismes pouvant causer les ITU en utilisant la PCR en temps réel pour amplifier le gène d’ARNr 23S des bactéries et 28S des levures. Grâce à une stratégie innovatrice de dénaturation des sondes, le pathogène peut être détecté et partiellement identifié. Une valeur seuil permettant de distinguer les échantillons positifs a été évaluée et correspond à un Cp de 26 (10[indice supérieur 5] ufc/mL). Un témoin interne est utilisé et permet de contrôler l’extraction et l’amplification. La méthode de dépistage a été éprouvée in vitro avec des souches de bactéries et levures, puis essayée avec quelques échantillons d’urines en provenance de la clinique. Puisque les résultats obtenus étaient très encourageants, l’analyse d’autres échantillons est importante pour faire une validation complète.