Three new eutardigrades, Doryphoribius longistipes sp. nov., Doryphoribius minimus sp. nov. and Doryphoribius tergumrudis sp. nov., are described from stream periphyton, rock lichen and cave soil and leaf litter samples collected in the limestone-based Cades Cove area of the Great Smoky Mountains National Park in eastern Tennessee, USA. Doryphoribius longistipes sp. nov. differs from other species mainly by the higher ratio of the secondary to the primary claw branches. Doryphoribius minimus sp. nov. differs from D. gibber, D. mariae and D. vietnamensis mainly by its smaller body size and different arrangement of dorsal gibbosities. Doryphoribius tergumrudis sp. nov. differs from D. evelinae by the absence of gibbosities and tubercles on legs I-III, the presence of lunulae and the absence of eyes. Additionally the three new species differ from other congeners by some other morphological and morphometric characters. Another species, Doryphoribius doryphorus Binda and Pilato, 1969, is a new record for Tennessee.

Based on the original species descriptions, a review of the genus Paramacrobiotus was conducted. We divided the genus into two subgenera, Microplacoidus subgen. nov. and Paramacrobiotus subgen. nov., based on the presence or absence of a microplacoid, characterized species within the genus based on seven different types of eggs. In a moss sample collected in Ecuador, Paramacrobiotus (Paramacrobiotus) spinosus sp. nov., was found. The new species differs from all species of the subgenus Parama crobiotus by the presence of richtersi type eggs and from other species by morphometric characters. Additionally, in the Ecuadorian material we found P. (Microplacoidus) magdalenae comb. nov., which is the first record of this species in Ecuador, we provide the full set of measurements for this species, not included in the original description. An additional new record is P. (M.) alekseevi comb. nov. found in Vietnam for the first time. After examining microscope slides from the Iharos' collection deposited in the Hungarian Natural History Museum, we prepared re-descriptions of P. (P.) csotiensis comb nov., P. (M.) submorulatus comb. nov. and P. (M.) wauensis comb. nov. Based on the morphological and morphometric characters of adults and eggs, we developed a diagnostic key to the genus Paramacrobiotus.

Durante la presente tesis doctoral se ha establecido la presencia de variabilidad interoperónica en el gen ARNr 16S de Aeromonas y observado que ésta afectaba a la taxonomía del género, limitando la identificación de A. caviae, A. media y A. veronii. La secuenciación del gen rpoD permitió identificar las cepas con variabilidad interoperónica a nivel de especie y reconocer 5 nuevas especies del género: A. fluvialis, A. taiwanensis, A. sanarelii, A. piscicola y A. rivuli. En colaboración con diversos hospitales españoles se han descrito los primeros aislados de A. aquariorum de origen extraintestinal, el segundo caso en adultos de síndrome urémico hemolítico (SUH) asociado a Aeromonas y se ha podido demostrar la presencia en este género del gen stx2 asociado al SUH. Se han revisado las características clínicas y microbiológicas de las infecciones de herida quirúrgicas relacionadas con Aeromonas. Finalmente, se ha descrito el primer caso de inducción in vivo de resistencia al imipenem en una cepa de A. veronii. Los resultados de esta tesis han dado lugar o contribuido a la publicación de 13 artículos científicos en revistas internacionales. / During this thesis has established the presence of inter-operon variability in 16S rRNA gene of Aeromonas and observed that it affected the taxonomy of the genus, limiting the identification of A. caviae, A. media and A. veronii. The rpoD gene sequences identified strains with variability at the species level and recognize 5 new Aeromonas species: A. fluvialis, A. taiwanensis, A. sanarelii, A. piscicola and A. rivuli. In collaboration with various Spanish hospitals have described the first isolation of A. aquariorum from extraintestinal origin, the second adult case of hemolytic uremic syndrome (HUS) associated with Aeromonas and demonstrated the presence of stx2 gene associated with HUS in Aeromonas. We reviewed the clinical and microbiological characteristics of surgical wound infections associated with Aeromonas. Finally, we described the first case of in vivo induction of resistance to imipenem in a strain of A. veronii. The results of this thesis have led or contributed to the publication of 13 scientific papers in international journals.

Made available in DSpace on 2016-08-29T15:33:47Z (GMT). No. of bitstreams: 1 tese_6327_Fernanda Tonini.pdf: 23203883 bytes, checksum: 1918c8ce1065fde9fefab194b21cebc8 (MD5) Previous issue date: 2013-02-22 / Pristocerinae são amplamente distribuídos desde os trópicos até as regiões temperadas do mundo, sendo atualmente compostos por 19 gêneros e cerca de 800 espécies. Estudos cladísticos mostram que Pristocerinae são monofiléticos, no entanto, a classificação de alguns gêneros ainda permanece inconsistente e as relações internas entre os mesmos não conhecidas. Caloapenesia Terayama, 1995, e Pseudisobrachium Kieffer, 1904 são os únicos gêneros de Pristocerinae que apresentam parâmeros duplos. Terayama (1996) analisou os gêneros de Pristocerinae cladisticamente, mas não investigou a fundo a presença de parâmeros duplos como uma possível sinapomorfia para a relação entre Caloapenesia e Pseudisobrachium. Terayama (1996) apresentou Pseudisobrachium como grupo-irmão de Protisobrachium e juntos formando um clado com Neoapenesia Terayama, 1995 suportado por apresentar notáulices ausentes ou indistintas e hipopígio com três hastes. Entretanto, estes caracteres são encontrados em outros gêneros da subfamília, e não foram investigados alguns caracteres diagnósticos destes três gêneros. No estudo de Terayama (1996), o hipopígio de Caloapenesia foi interpretado como tendo apenas uma haste, embora o próprio autor do gênero, em 1995, tenha ilustrado as hastes laterais inconspícuas, indicando que este gênero compartilharia uma das sinapomorfias do clado rotisobrachium+Pseudisobrachium+Neoapenesia. Neoapenesia também apresenta uma projeção apical no basiparâmero, que pode ser homóloga ao segundo parâmero de Caloapenesia e Pseudisobrachium, mas este caráter não havia sido descrito para o gênero e não foi analisado na única análise cladística feita para a subfamília. Além da falta de delimitação cladística, a maioria dos gêneros de Pristocerinae tem menos de dez espécies descritas, e é conhecida restrita às regiões Afrotopical e Oriental. Caloapenesia tem atualmente apenas três espécies válidas descritas e é restrito à região Oriental. Muitos de seus caracteres considerados diagnósticos são compartilhados por outros gêneros, e alguns caracteres não haviam sido observados para o táxon. Os objetivos deste trabalho foram revisar taxonomicamente Caloapenesia; descrever um gênero novo com parâmero duplo; investigar a homologia do parâmero duplo dos três gêneros de Pristocerinae que o possuem; investigar a homologia entre a projeção de Neoapenesia com o parâmero duplo de Caloapenesia e Pseudisobrachium; testar a monofilia e investigar as possíveis relações de parentesco destes gêneros.

A utilização de produtos naturais para a terapêutica do câncer foi iniciada com a actinomycina D, obtida a partir de culturas de Streptomyces e desde então, a busca por compostos bioativos de origem natural constitui uma importante linha de pesquisa. Estima-se que aproximadamente 60% dos agentes antineoplásicos, introduzidos para a terapia do câncer nas últimas décadas, tem origem vegetal ou microbiana. Dentre os micro-organismos proeminentes para produção de compostos ativos, as actinobactérias se destacam pela versatilidade metabólica, praticidade para cultivo in vitro e eficiência para produção de compostos com atividade anticâncer. Em seu último relatório, a Organização Mundial da Saúde reportou 8,8 milhões de mortes em decorrência de câncer, no ano de 2017. O índice representa um em cada seis óbitos em todo o mundo, sendo mais expressivo em países de média e baixa renda. Vale ressaltar que avanços significativos foram alcançados nos últimos anos para o tratamento de leucemia aguda infantil e tumores derivados de células germinais. Contudo, tumores sólidos de pulmão, próstata, mama e cólon ainda representam altos índices de mortalidade. Frente a isso, torna-se evidente a necessidade de identificar e desenvolver estratégias para o tratamento da doença. Com intuito de acessar novos recursos microbianos com potencial biotecnológico, a prospecção avança para áreas pouco exploradas, como por exemplo, o Continente Antártico. A Antártica foi o último dos continentes a ser acessado pelo homem e apresenta características edafoclimáticas favoráveis ao endemismo. Em vista da problemática e da potencialidade do Continente Antártico, os recursos microbiológicos associados à rizosfera de Deschampsia antarctica Desv. foram acessados e avaliados para a produção de compostos com propriedade antitumoral. Em resultado foram obtidos 42.528 clones metagenômicos e 72 linhagens de actinobactérias, dentre as quais Streptomyces sp. CMAA 1527, que apresentou pronunciada atividade antiproliferativa in vitro, para tumores de mama, pulmão, rim e sistema nervoso central, através da produção de cinerubina B. A análise taxonômica das actinobactérias isoladas revelou a presença de linhagens com baixo índice de similaridade, com as linhagens tipo conhecidas, o que pode significar a presença de novas espécies para os gêneros Nocardia, Rhodococcus e Streptomyces, reconhecidos pela capacidade de produzir metabólitos ativos e enzimas de interesse biotecnológico. A análise taxonômica polifásica da linhagem CMAA 1533 possibilitou a descrição da espécie Rhodococcus psychrotolerans sp. nov. (TaxoNumber TA00191; NRRL B-65465T = DSM 104532T), grupo bacteriano promissor como agente de biorremediação e produção de compostos bioativos. Com isso, o Continente Antártico foi considerado um ambiente promissor para a busca de novos micro-organismos, dentre eles actinobactérias, eficientes na produção de compostos antitumorais e outras substâncias com potencial biotecnológico. / The use of natural products for cancer therapy was initiated with the actinomycin D, obtained from Streptomyces. Since then, the search of bioactive from natural sources represent an essential line of research. It is estimated that approximately 60% of the antineoplasic agents inserted for the cancer therapy in recent decades have vegetal and microbial origin. Among the prominent microorganisms used to produce active compounds, actinobacterias are known by their metabolical versatility, convenience related to in vitro culture, and efficiency on the production of anticancer compounds. The Health World Organization, on its last review, reported 8.8 million of deaths in 2017, caused by cancer. Those numbers represent one out of six deaths worldwide, being more expressive in middle and low income countries. It is worth pointing out that meaningful advances were established in recent years for the treatment of childhood acute leukemia and germ cell-derived tumors. However, solid tumors of the lung, prostate, breast and colon still represent high mortality rates. For this reason, it is necessary to identify and develop strategies for the treatment of the disease. With the aim of accessing new microbial resources that contain biological potential, the prospection advance to areas barely explored, such as the Antarctic Continent. Antarctica was the last of the continents to be accessed by man and presents edaphoclimatic characteristics favorable to endemism. In light of the problematic and the potentiality of the Antarctic Continent, the microbiological resources associated with the rhizosphere of Deschampsia antarctica Desv. were accessed and evaluated for the production of compounds with antitumor properties. The results obtained had shown 42,528 metagenomic clones and 72 strains of actinobacteria, among them Streptomyces sp. CMAA 1527, which had presented anti-proliferative activity in vitro to breast, lung, kidney and central nervous system tumors, through the production of cinerubin B. The taxonomic analysis of the actinobacteria isolated revealed the presence of strains with low rate of similarity, with known type strains, which may mean the presence of new species for the genera Nocardia, Rhodococcus and Streptomyces, recognized for the ability to produce active metabolites and enzymes of biotechnological interest. The polyphasic approach of the CMAA 1533 strain made possible the description of the species Rhodococcus psychrotolerans sp. nov. (TaxoNumber TA00191; NRRL B-65465T = DSM 104532T), promising bacterial group as a bioremediation agent and production of bioactive compounds. As a result, the Antarctic Continent was considered a promising environment to search new microorganisms, among them, the actinobacteria, which is efficient on the production of antitumor compounds and other substances with biotechnological potential.

A utilização de produtos naturais para a terapêutica do câncer foi iniciada com a actinomycina D, obtida a partir de culturas de Streptomyces e desde então, a busca por compostos bioativos de origem natural constitui uma importante linha de pesquisa. Estima-se que aproximadamente 60% dos agentes antineoplásicos, introduzidos para a terapia do câncer nas últimas décadas, tem origem vegetal ou microbiana. Dentre os micro-organismos proeminentes para produção de compostos ativos, as actinobactérias se destacam pela versatilidade metabólica, praticidade para cultivo in vitro e eficiência para produção de compostos com atividade anticâncer. Em seu último relatório, a Organização Mundial da Saúde reportou 8,8 milhões de mortes em decorrência de câncer, no ano de 2017. O índice representa um em cada seis óbitos em todo o mundo, sendo mais expressivo em países de média e baixa renda. Vale ressaltar que avanços significativos foram alcançados nos últimos anos para o tratamento de leucemia aguda infantil e tumores derivados de células germinais. Contudo, tumores sólidos de pulmão, próstata, mama e cólon ainda representam altos índices de mortalidade. Frente a isso, torna-se evidente a necessidade de identificar e desenvolver estratégias para o tratamento da doença. Com intuito de acessar novos recursos microbianos com potencial biotecnológico, a prospecção avança para áreas pouco exploradas, como por exemplo, o Continente Antártico. A Antártica foi o último dos continentes a ser acessado pelo homem e apresenta características edafoclimáticas favoráveis ao endemismo. Em vista da problemática e da potencialidade do Continente Antártico, os recursos microbiológicos associados à rizosfera de Deschampsia antarctica Desv. foram acessados e avaliados para a produção de compostos com propriedade antitumoral. Em resultado foram obtidos 42.528 clones metagenômicos e 72 linhagens de actinobactérias, dentre as quais Streptomyces sp. CMAA 1527, que apresentou pronunciada atividade antiproliferativa in vitro, para tumores de mama, pulmão, rim e sistema nervoso central, através da produção de cinerubina B. A análise taxonômica das actinobactérias isoladas revelou a presença de linhagens com baixo índice de similaridade, com as linhagens tipo conhecidas, o que pode significar a presença de novas espécies para os gêneros Nocardia, Rhodococcus e Streptomyces, reconhecidos pela capacidade de produzir metabólitos ativos e enzimas de interesse biotecnológico. A análise taxonômica polifásica da linhagem CMAA 1533 possibilitou a descrição da espécie Rhodococcus psychrotolerans sp. nov. (TaxoNumber TA00191; NRRL B-65465T = DSM 104532T), grupo bacteriano promissor como agente de biorremediação e produção de compostos bioativos. Com isso, o Continente Antártico foi considerado um ambiente promissor para a busca de novos micro-organismos, dentre eles actinobactérias, eficientes na produção de compostos antitumorais e outras substâncias com potencial biotecnológico. / The use of natural products for cancer therapy was initiated with the actinomycin D, obtained from Streptomyces. Since then, the search of bioactive from natural sources represent an essential line of research. It is estimated that approximately 60% of the antineoplasic agents inserted for the cancer therapy in recent decades have vegetal and microbial origin. Among the prominent microorganisms used to produce active compounds, actinobacterias are known by their metabolical versatility, convenience related to in vitro culture, and efficiency on the production of anticancer compounds. The Health World Organization, on its last review, reported 8.8 million of deaths in 2017, caused by cancer. Those numbers represent one out of six deaths worldwide, being more expressive in middle and low income countries. It is worth pointing out that meaningful advances were established in recent years for the treatment of childhood acute leukemia and germ cell-derived tumors. However, solid tumors of the lung, prostate, breast and colon still represent high mortality rates. For this reason, it is necessary to identify and develop strategies for the treatment of the disease. With the aim of accessing new microbial resources that contain biological potential, the prospection advance to areas barely explored, such as the Antarctic Continent. Antarctica was the last of the continents to be accessed by man and presents edaphoclimatic characteristics favorable to endemism. In light of the problematic and the potentiality of the Antarctic Continent, the microbiological resources associated with the rhizosphere of Deschampsia antarctica Desv. were accessed and evaluated for the production of compounds with antitumor properties. The results obtained had shown 42,528 metagenomic clones and 72 strains of actinobacteria, among them Streptomyces sp. CMAA 1527, which had presented anti-proliferative activity in vitro to breast, lung, kidney and central nervous system tumors, through the production of cinerubin B. The taxonomic analysis of the actinobacteria isolated revealed the presence of strains with low rate of similarity, with known type strains, which may mean the presence of new species for the genera Nocardia, Rhodococcus and Streptomyces, recognized for the ability to produce active metabolites and enzymes of biotechnological interest. The polyphasic approach of the CMAA 1533 strain made possible the description of the species Rhodococcus psychrotolerans sp. nov. (TaxoNumber TA00191; NRRL B-65465T = DSM 104532T), promising bacterial group as a bioremediation agent and production of bioactive compounds. As a result, the Antarctic Continent was considered a promising environment to search new microorganisms, among them, the actinobacteria, which is efficient on the production of antitumor compounds and other substances with biotechnological potential.

This is the first survey of subtidal marine tardigrades from the Bahamas, and we compare our results with earlier studies of Bahamian intertidal habitats. In 2011 and 2014 we collected 60 subtidal sand samples from Bimini, the Berry Islands, New Providence, Eleuthera, and the Exumas. We found 11 species only one of which, Dipodarctus subterraneus (Renaud- Debyser, 1959), had been found in the previous intertidal Bahamian collections. Thus, 10 species are new records for the Bahamas, and four of these are new to science. We describe two of the new species (Archechiniscus bahamensis sp. nov. and Anisonyches eleutherensis sp. nov.). Additional material is needed for a conclusive identification of the undescribed species of Batillipes and Florarctus. Based on examination of the holotype of Anisonyches diakidius Pollock, 1975, we redescribe this species and give an emended diagnosis of the genus and the species. We also provide new keys to the spe-cies in the genera Archechiniscus and Anisonyches. A Chao2 species richness estimate indicated that our sampling effort was extensive and probably recovered most common species. Correlations of abundance and diversity indices with regard to depth, distance to nearest shore, and sediment grain size were not significant.

Background. The Cysteine-rich protein, Connective tissue growth factor, and Nephroblastoma overexpressed protein (CCN) family of matricellular proteins are signaling molecules found in the extracellular space, which can have pro-angiogenic, anti-inflammatory and anti-fibrotic properties. Their expression and role in repair and remodeling after myocardial infarction (MI) remains to be better elucidated. In this study, the age-associated expression of Nov (CCN3) and Wisp1 (CCN4) were examined post-MI in mice. Methods and Results. In vivo, MI was induced in young (6 week) and old (12-14 months) mice. Cardiac function was assessed by echocardiography, showing that LVEF was reduced in old mice (33.9%) at 14 days post-MI compared to young mice (43.9%; p=0.002). RT-qPCR analysis of harvested myocardial tissue revealed that mRNA expression of several matricellular proteins in healthy tissue was decreased by 2.5- to 5-fold in old compared to young mice (p=0.03 for Nov, p=0.04 for Wisp1, p=0.0002 for TnC, p=0.04 for TSP-1). Post-MI, mRNA expression of Nov was reduced in the infarct (by up to 13-fold; p<0.03) and border zone (by up to 16-fold; p<0.002) in old compared to young mice. Nov and Wisp1 protein expression was also reduced in old compared to young mice in the infarct and border zones; specifically, for Nov in the infarct zone (p=0.01) and the border zone (p=0.009) at 2 days post-MI and for Wisp1 in the infarct zone at 2 days (p=0.0003) and 14 days (p=0.003), along with 7 days post-MI in the border zone (p=0.0003). To identify possible sources of matricellular proteins, in vitro culture experiments were performed. The expression of Nov protein was increased (1.9-fold; p=0.006) in TGF-B stimulated cardiac fibroblasts after 48h, as was the expression of the myofibroblast marker a-SMA (1.7-fold; p=0.035). Wisp1 mRNA expression was increased (4.5-fold; p=0.03) in stimulated cardiac fibroblasts after 48h in a hypoxic environment. There was also a trend for increased mRNA expression of Nov (p=0.118) and Wisp1 (p=0.121) in M2 macrophages. Cardiac fibroblasts treated with Nov+TGF-B exhibited greater proliferation (by 29%; p0.01), as did those treated with Wisp1+TGF-B (by 16%; p<0.05). Treatment with Nov or Wisp1 led to an increase in viability of cardiac fibroblasts both in the presence (Nov; p=0.0004, Wisp1; p=0.01) and absence of TGF-B (Nov; p=0.0005, Wisp1; p=0.003). Summary. There is an age-associated difference in the expression of matricellular proteins Nov and Wisp1 between both healthy and MI mice. In vitro studies suggest that cardiac fibroblasts may produce Nov and Wisp1 upon their activation to myofibroblasts. The presence of these proteins was also shown to increase the proliferation and viability of fibroblasts. Therefore, reduced levels of Nov and Wisp1 in old mice may negatively affect the repair and remodeling process post-MI compared to young mice. A better understanding of Nov and Wisp1 function in aging and post-MI repair may help identify novel therapeutic targets for limiting damage post-MI and improving repair and heart function.

The present study documents the first confirmed fossil record of Sinomenium in Menispermaceae, Sinomenium macrocarpum sp. nov., from the recently discovered Gray Fossil Site in Tennessee, southeastern USA. The fossil species is represented by more than 120 endocarps, all of which are characterized by their horseshoe-shaped form and occurrence of highly ornamented protuberances on both dorsal and lateral crests. A combination of their relatively large size and highly developed of protuberances on the surface of endocarp warrants the new species. The new species is justified through a detailed comparison with the related and published extant and fossil species. The discovery of the genus in North America appears to support that the Gray site in southern Appalachian region represented a forest refugium during the late Neogene when the global cooling was intensified and grasslands were globally expanded.

As part of the All Taxa Biodiversity Inventory (http://www.dlia.org), we are conducting a large-scale multihabitat inventory of tardigrades in the Great Smoky Mountains National Park (GSMNP) in Tennessee and North Carolina, USA. Here we report our findings for the genus Macrobiotus (Eutardigrada, Macrobiotidae). Two new species, Macrobiotus martini sp. nov. and Macrobiotus halei sp. nov., are described from moss, lichen, soil and leaf litter samples. Macrobiotus martini sp. nov. differs from all other species of the Macrobiotus hufelandi group by having a very simple buccal armature without bands of teeth, very large elliptical cuticular pores, unique characteristics of the egg, and other morphometric characters. Macrobiotus halei sp. nov. differs from other species in the Macrobiotus richtersi group by having very small cuticular tubercles, well-developed macroplacoids, unique characteristics of the egg, and other morphometric characters. Seven additional Macrobiotus species have been identified in the GSMNP: M. harmsworthi Murray, 1907, M. hufelandi Schultze, 1834, M. islandicus Richters, 1904, M, montanus Murray, 1910, M. pallarii Maucci, 1954, M. recens Cuénot, 1932, and M. tonollii Ramazzotti, 1956. All of these except M. harmsworthi are new records for the national park. In addition, M. pallarii is a new record for North Carolina, M. islandicus is a new record for North Carolina and Tennessee, M. montanus is a new record for the eastern USA, and M. recens is a new record for the USA.