Several biomarkers (cell viability and phagocytosis) based on earthworm (Lumbricus terrestris) immune cells (coelomocytes), together with whole-worm mortality (LC/LD50's), were used to assess a bioremediation attempt to reduce pentachlorophenol (PCP) toxicity in a former wood processing hazardous waste site (HWS).

This research evaluated the potential of activation and attachment, as sequential companion biomarkers of phagocytosis by earthworm, Lumbricus terrestris, immunoactive coelomocytes for use in immunotoxicology. The potential was assessed by exposing earthworms to sublethal concentrations of CuSO4 and Arochlor 1254®, chemicals used as reference or standard immunotoxicants.

Equinodermos são bem conhecidos pelas suas grandes capacidades regenerativas, principalmente em processos mais complexos como a reposição de membros e órgãos. Porém pouco se sabe sobre processos aparentemente mais simples como a cicatrização. Tem sido descrito que um dos principais mecanismos envolvidos neste evento é o remodelamento da matriz extracelular e que existem células responsáveis por isto. Entretanto, vários detalhes do processo e do exato papel destas células ainda não são muito claros. Neste trabalho, foram estudados os estágios do processo de cicatrização em Holothuria grisea Selenka, 1867 (Holothuriidae). A lesão foi induzida através de uma incisão perfurando a parede do corpo do animal até alcançar a cavidade celomática. Através de análises histológicas, foram acompanhadas as mudanças estruturais e os mecanismos celulares que ocorrem no intervalo de doze horas a trinta dias após a injúria. Foi observado que houve um rápido fechamento da ferida através da síntese de novas fibras de colágeno, e que fibroblastos e duas populações de esferulócitos estão envolvidos neste processo. São discutidas as prováveis funções destes tipos celulares e possíveis diferenças funcionais entre os esferulócitos. / Echinoderms are well known by their regenerative capabilities, mostly in complex events such as the reconstruction of internal organs or entire body parts. However, the knowledge on apparently simpler processes such as wound healing is relatively scarce. It has been described that extracellular matrix remodeling is the main mechanism, and it involves participation of different cells populations. However, the specific function of these cell types is still under debate. In this work, the wound healing process in Holothuria grisea Selenka, 1867 (Holothuriidae) was studied. The lesion was made by an incision through the body wall up to the coelomic cavity. The local changes in cell types and the reorganization of the extracellular matrix were accompanied from 12 hours to 30 days after the injury. Soon after the wound, the organism responded with a localized contraction, followed by cell migration and synthesis of new collagen fibers. Four different cell types were observed, including two different types of spherulocytes. The probable functions of these cells are discussed.

Equinodermos são bem conhecidos pelas suas grandes capacidades regenerativas, principalmente em processos mais complexos como a reposição de membros e órgãos. Porém pouco se sabe sobre processos aparentemente mais simples como a cicatrização. Tem sido descrito que um dos principais mecanismos envolvidos neste evento é o remodelamento da matriz extracelular e que existem células responsáveis por isto. Entretanto, vários detalhes do processo e do exato papel destas células ainda não são muito claros. Neste trabalho, foram estudados os estágios do processo de cicatrização em Holothuria grisea Selenka, 1867 (Holothuriidae). A lesão foi induzida através de uma incisão perfurando a parede do corpo do animal até alcançar a cavidade celomática. Através de análises histológicas, foram acompanhadas as mudanças estruturais e os mecanismos celulares que ocorrem no intervalo de doze horas a trinta dias após a injúria. Foi observado que houve um rápido fechamento da ferida através da síntese de novas fibras de colágeno, e que fibroblastos e duas populações de esferulócitos estão envolvidos neste processo. São discutidas as prováveis funções destes tipos celulares e possíveis diferenças funcionais entre os esferulócitos. / Echinoderms are well known by their regenerative capabilities, mostly in complex events such as the reconstruction of internal organs or entire body parts. However, the knowledge on apparently simpler processes such as wound healing is relatively scarce. It has been described that extracellular matrix remodeling is the main mechanism, and it involves participation of different cells populations. However, the specific function of these cell types is still under debate. In this work, the wound healing process in Holothuria grisea Selenka, 1867 (Holothuriidae) was studied. The lesion was made by an incision through the body wall up to the coelomic cavity. The local changes in cell types and the reorganization of the extracellular matrix were accompanied from 12 hours to 30 days after the injury. Soon after the wound, the organism responded with a localized contraction, followed by cell migration and synthesis of new collagen fibers. Four different cell types were observed, including two different types of spherulocytes. The probable functions of these cells are discussed.

Submitted by Vasti Diniz (vastijpa@hotmail.com) on 2017-09-08T12:11:24Z No. of bitstreams: 1 arquivototal.pdf: 1391577 bytes, checksum: e8bbd0db33d228b40d69b547aed31f9c (MD5) / Made available in DSpace on 2017-09-08T12:11:24Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 1391577 bytes, checksum: e8bbd0db33d228b40d69b547aed31f9c (MD5) Previous issue date: 2016-02-29 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Human activities have caused climate changes and altered the salinity of the oceans. Salinity is one of the factors that limit the distribution and the survival of marine organisms. Coelomocytes are the immune system cells of the echinoderms and have been studied as biomarkers in stress situations. The aim of the present study was to investigate the effect of the salinity in the immune system cells of the tropical sea urchin Echinometra lucunter. Animals were collected in João Pessoa coast (Brazilian Northeast). Animals or coelomocytes were exposed to different salinity (25‰ to 45‰) and phagocytic parameters, production of reactive oxygen species (ROS), mitochondrial activity and ABC transporter activity analyzed. The phagocytic parameters did not change when animals or cells were exposed to low or high salinity in any time intervals monitored. However, our data showed an increase in the coelomocytes concentration when animals were exposed to 25‰. ROS levels were higher when cells were incubated at 25‰ and lower when cells were cultured at 45‰. We noted a loss of the mitochondrial inner membrane potential when coelomocytes were incubated at 45‰. The activity of ABC transporters decreased when cells were incubated at low salinity and increased when cells were incubated at high salinity. Our work shows that the immune system of the tropical sea urchins E. lucunter tolerates salinity changes from 25‰ to 45‰ and suggests two cellular parameters (ROS levels and ABC transporters activity) as potential biomarkers on the monitoring of the impact of environmental salinity changes. / As atividades humanas têm causado mudanças climáticas e alterado a salinidade dos oceanos. A salinidade é um dos fatores que limitam a distribuição e sobrevivência de organismos marinhos. Celomócitos são as células do sistema imune dos equinodermos e têm sido estudados como biomarcadores em situações de estresse. O objetivo do presente estudo foi investigar o efeito da salinidade em celomáticos do ouriço-do-mar tropical Echinometra lucunter. Os animais foram coletados na costa de João Pessoa (Nordeste do Brasil). Os animais ou os celomócitos foram expostos a diferentes salinidades (25‰ e 45‰) e parâmetros fagocíticos, produção de espécies reativas de oxigênio (ROS), atividade mitocondrial e atividade dos transportadores ABC analisados. Os parâmetros fagocíticos não alteraram quando os animais ou as células foram expostos a 25‰ ou 45‰ nos intervalos de tempo monitorados. Porém, foi observado um aumento na concentração de celomócitos quando os animais foram expostos a 25‰. Os níveis de ROS foram maiores quando as células foram incubadas a 25‰, e menores quando as células foram cultivadas a 45‰. Foi observada uma perda do potencial de membrana mitocondrial interna quando os celomócitos foram incubados a 45‰. A atividade dos transportadores ABC diminuiu quando as células foram incubadas a 25‰ e aumentou quando as células foram incubadas a 45‰. O presente trabalho demonstra que o sistema imune do ouriço-do-mar E. lucunter tolera mudanças de salinidade (25‰ até 45‰), e sugere dois parâmetros celulares (níveis de ROS e atividade de transportadores ABC) como potenciais biomarcadores no monitoramento de mudanças na salinidade ambiental.

STORAGE CELLS AND THEIR ROLE IN TARDIGRADE PHYSIOLOGY Abstract Tardigrades possess remarkable tolerance to numerous stress conditions (e.g. almost complete desiccation, exposure to very low sub-zero temperature, heat stress and even exposure to space in low Earth orbit). Indeed, they are among the most radiation-resistant multi-cellular organisms. The body cavity of tardigrades is filled with the storage cells (SC). Their role in anhydrobiosis has been discussed. The main objectives of this work were to analyse (i) the occurrence of mitosis in SC, (ii) the factors constraining anhydrobiotic survival, and (iii) the general ultrastructure of SC and their ultrastructure concerning the stress conditions. Our model species, R. cf. coronifer is one of the most extensively studied tardigrades concerning anhydrobiosis. Comprehensive histochemical techniques were used in combination with SEM, TEM, and confocal microscopy. First, mitotic divisions of tardigrade SC occur with a higher frequency in juveniles than in adults and correlate with animal growth. Mitosis is more frequent in moulting tardigrades, but the overall mitotic index is low. Furthermore, tardigrades of R. cf. coronifer can survive the maximum of 6 repeated desiccation cycles with significantly declining survival rate with repeated desiccations and...