Freshwater sponges (Porifera, Demospongiae) indicators of some coastal habitats in South America: redescriptions and key to identification

Surveys for freshwater sponges were performed at several water bodies at sandy environments along a north-south direction of particularly the Brazilian coastal line. The results allowed for the distinction of four different species-specific environments along this coastal border. The main fact considered was the dominant or the sole recurrent occurrence of a single sponge species at one particular habitat. The first one is that of the lagoonal mesohaline habitats at the tropical and subtropical realms, indicated by Spongilla alba Carter, 1849. The second one refers to shallow ponds among dunes at the tropical area indicated by Corvoheteromeyenia heterosclera (Ezcurra de Drago, 1974). The third one is that of also shallow ponds close to the dune belt at the temperate region indicated by Racekiela sheilae (Volkmer-Ribeiro, De Rosa-Barbosa & Tavares, 1988). The fourth one is that of organically enriched environments, at the marginal areas of lagoons and mouth of small rivers, evolving towards freshwater muddy ponds and coastal swamps, not far from the ocean border: Ephydatia facunda Weltner, 1895 is the species to occupy this habitat with almost exclusiveness. The above species are thus proposed as indicators of such habitats and have their descriptions improved and that of their environments summarized. A taxonomic key based on the spicules of the four species is proposed. The results presented aim to contribute to the identification of spicules of these sponges in sediment columns recovered at the Brazilian and South American coastal area. Determination of paleo ocean borders are a present issue of upmost importance in what respects projections of timing and fluctuations of ascending/regressing sea levels.

Freshwater sponges (Porifera, Demospongiae) in a benthic filter feeding community at the Guanacaste Dry Forest, Costa Rica

The freshwater sponges Trochospongilla variabilis Bonetto & Ezcurra de Drago (1973), Radiospongilla crateriformis (Potts, 1882), Spongilla cenota Penney & Racek (1968) and Corvoheteromeyenia heterosclera (Ezcurra de Drago, 1974) compose with the sphaerid bivalve Eupera cubensis (Prime, 1865) and several Phylactolaemata bryozoans a benthic filter feeding community living in seasonal lentic and lotic habitats with high Particulate Organic Carbon (POC), low conductivity and acid pH within the Costa Rica Dry Forest biome. The sponge specimens gathered led to the re-description of the four species.

Acanthotylotra alvarengai (Porifera, Demospongiae) new genus and species of sponge from Tocantins river, Pará State, Brazil

Acanthotylotra a new monotypic genus of freshwater sponge is defined. Its sole species A. alvarengai sp. nov. is described based upon microscopic specimens recovered from the Tocantins river, Pará State, Brazil, when the deep rocky substrate was temporarily exposed downstream from the dam at the time it was closed for lake formation. A unique set of megascleres, allied to the paucity of spongin, the renieroid skeleton and the fact that gemmules or new specimens remain undetected, call for the proposition of a new monospecific genus to be retained as incertae sedis until new larger and probably gemmuliferous specimens come to be found.

Dosilia (Porifera, Demospongiae) redefined

Dosilia plumosa (Carter, 1849), type species of the genus, and D. brouni (Kirkpatrick, 1906), with distribution respectively in the Oriental and Ethiopic regions, are revised based on a SEM analysis of spicules, gemmules and skeletal structure. The lectotype here designated for D. plumosa is illustrated as well as the holotype by monotypy determined for D. brouni. Dosilia palmeri (Potts, 1885) and D. radiospiculata (Mills, 1888) distributed in the Nearctic and Neotropical regions and D. pydanieli Volkmer-Ribeiro, 1992, found in the Neotropical region, are revised based on a SEM analysis of spicules, gemules and skeletal structure. The holotype by monotypy is determined for D. radiospiculata. Heteromeyenia plumosa Weltner,1895 is synonymyzed with D. radiospiculata. Upon the revision of its five species, the genus is redefined and a key presented.

Species of Oukuriella Epler (Diptera, Chironomidae) inside freshwater sponges in Brazil

Larvae of Oukuriella Epler, 1986 (Diptera, Chironomidae) inside freshwater sponges are reported for the first time in Brazil.

Metabólitos secundários das esponjas Aplysina fistularis e Dysidea sp. e atividade antituberculose da 11-cetofistularina-3

The present investigation reports the isolation of aeroplysinin-2, 2-(3,5-dibromo-4-methoxyphenyl)-N,N,N-trimethyletanamonium, 7,9-dibromo-10-hydroxy-8-methoxy-1-oxa-2-azaspiro[4.5]deca-2,6,8-trien-3-carboxylic acid and its methyl ester, 11-oxoaerothionin, aerothionin, 11-keto-12-hydroxyaerothionin, 11-ketofistularin-3 and fistularin-3 from Aplysina fistularis, as well as of furodysinin lactone and 9α,11α-epoxicholest-7-en-3β,5α,6α,10-tetrol-6-acetate from Dysidea sp. Although the extracts of both sponges displayed antituberculosis activity, only 11-ketofistularin-3 isolated from A. fistularis displayed antimycobacterial activity against Mycobacterium tuberculosis H34Rv, with MIC at 16 μg/mL and SI of 40, a result that reinforce that fistularin-3 derivatives are interesting leads for the development of antituberculosis drugs.

Spongia (Heterofibria) catarinensis sp. nov. (Porifera, Spongiidae) no litoral de Santa Catarina, Brasil

Uma nova espécie, Spongia (Heterofibria) catarinensis, é descrita para a Ilha das Aranhas (27º29'077''S, 48º21'380''W), Estado de Santa Catarina, Brasil. Difere das outras espécies do gênero do Atlântico sudoeste pela morfologia externa e arquitetura. A nova espécie é caracterizada por apresentar forma massiva, incrustante com projeções lobulares; fibras primárias medindo 60-100 µm; fibras secundárias 11,5-69 µm; fibras pseudoterciárias 2,3-23 µm e distância entre fibras primárias 391-920 µm; distância entre fibras secundárias/pseudoterciárias 92-575 µm; fibras secundárias/pseudoterciárias constituindo malhas poligonais com 30-700 µm de diâmetro. A espécie pertence ao subgênero Heterofibria Cook & Bergquist, 2001 por apresentar uma clara dicotomia de suas fibras.

Soluções nutritivas para cultivo e produção de frutanos em plantas de Vernonia herbacea

O crescimento limitado de rizóforos de Vernonia herbacea (Asteraceae) em solução de Hoagland levou à necessidade de estabelecer uma solução nutritiva para o cultivo dessa planta, visando ao incremento da biomassa de seus rizóforos ricos em frutanos. Essa solução (denominada Vernonia), constituída de Ca(NO3)2.4H2O 2,5 mmol L-1, KNO3 2,3 mmol L-1 , KH2PO4 0,52 mmol L-1, Mg(NO3)2.6H2O 1,7 mmol L-1 e Na2SO4 1,3 mmol L-1, foi comparada com a de Hoagland nas forças iônicas de 50%, 100% e 200%. Foram realizadas duas avaliações para análise de crescimento e conteúdo de frutanos. As plantas não sobreviveram até os dois meses na solução de Hoagland 200%. A solução Vernonia diluída duas vezes (50%) foi a mais eficiente para o incremento de massa seca dos rizóforos e produção de frutanos por planta. Maior crescimento da parte aérea foi verificado nas soluções de Hoagland e Vernonia 100%. Em comparação com a solução de Hoagland, a solução Vernonia é mais pobre em macronutrientes, confirmando a hipótese de que plantas adaptadas a solos oligotróficos são menos exigentes em nutrientes minerais.

Metabólitos secundários dos nudibrânquios Tambja stegosauriformis, Hypselodoris lajensis e Okenia zoobotryon e dos briozoários Zoobotryon verticillatum e Bugula dentata da costa do Brasil

The chemical investigation of the MeOH extract from the bryozoan B. dentata MeOH yielded tambjamines A (1), C (3), D (4), K (6), aldehyde 8 and the new tambjamine J1(9), while the extract of its predator, the nudibranch Tambja stegosauriformis, yielded tambjamines C and K, along with aldehyde 8. Furodisinin lactone (11) was isolated from the nudibranch Hypselodoris lajensis, a compound previously isolated from Dysidea sponges. The alkaloid 2,5,6-tribromo-N-methylgramine (12) was isolated from the nudibranch Okenia zoobotryon and from its prey, the bryozoan Zoobotryon verticillatum, the only source of 12 previously known.

Inulin production by Vernonia herbacea as influenced by mineral fertilization and time of harvest

The underground organs of Vernonia herbacea (Vell.) Rusby, known as rhizophores, acumulate 80% of their dry mass as fructans of the inulin type. In view of the growing industrial use of fructans as dietetic and general food products, and of their medical application, the present investigation aimed at evaluating the effect of mineral fertilization and period of cultivation on the production of these carbohydrates in field trials. Plants used in the experiments were obtained by vegetative propagation from rhizophores collected from plants growing in natural areas of the cerrado, and cultivated for two years. Fertilization consisted of N:P2O5:K2O (80:200:150 kg.ha-1) plus 80 kg.ha-1 nitrogen as dressing. Soil fertilization did not stimulate biomass or inulin production, but in the second year of cultivation a dramatic gain in biomass and inulin was detected in both treated and control plants. Inulin production varied from 113 to 674 kg.ha-1 which corresponds to 43% of the rhizophore dry mass. The composition of fructans was not altered by fertilization, although treated plants had a higher proportion of sucrose and fructans with degree of polymerization 3-8 in the second year of cultivation. The results identify this species as a fructan source similar to other commercial crops and recommend further agronomic studies, aimed at increasing the production of this polysaccharide.

Fructan variation in the rhizophores of Vernonia herbacea (Vell.) Rusby, as influenced by temperature

The influence of climatic variations on fructan content in tropical regions is not well known. The present study deals with the effects of temperature on fructan contents in rhizophores of plants of Vernonia herbacea, a native species from the Brazilian cerrado vegetation. Intact plants and fragmented rhizophores were subjected to different temperatures under natural and controlled environmental conditions. Rhizophores of plants in pre-dormant stage (aerial parts showing some yellowish leaves) presented higher fructan content at 5oC than those kept at 25oC, whereas in dormant plants (aerial parts absent) temperature treatments did not affect fructan contents. Fragmented rhizophores obtained from dormant plants presented higher levels of fructo-polysaccharides at the end of the experiment than at the beginning of the treatment, regardless of the temperature they were stored, whereas fragments obtained from vegetative plants showed a decrease in fructan content under the same treatments. It was concluded that variations observed in fructan contents are related to the phenological state of the plants prior to the treatment rather than to extraneous temperatures they are subjected to during this stage.

Crescimento e conteúdo de frutanos em plantas de Vernonia herbacea (Vell.) Rusby submetidas à adubação nitrogenada

Vernonia herbacea acumula altas concentrações de frutanos do tipo inulina em seus órgãos subterrâneos de reserva, denominados rizóforos. Foi estudada a influência da adubação nitrogenada no crescimento dessa espécie e na produção de frutanos. Plantas obtidas a partir de fragmentos de rizóforos foram cultivadas em área de cerrado onde receberam três doses de nitrogênio: 6, 12 e 24 kg N.ha-1 na forma de (NH4)2SO4. Plantas jovens, com seis meses de idade, mostraram aumento em altura, área foliar e massa seca da parte aérea e dos rizóforos em resposta ao aumento da dose de nitrogênio. Por outro lado, o teor de frutanos decresceu com o aumento da concentração do nitrogênio. Após doze meses de cultivo, o crescimento das plantas foi estimulado apenas nas doses entre 6 e 12 kg.ha-1 de nitrogênio. Os resultados mostraram, também, que o teor de frutanos em plantas jovens está correlacionado negativamente tanto ao crescimento quanto às concentrações crescentes de nitrogênio. Já as plantas tratadas com 24 kg N.ha-1 chegaram a acumular 6,0 g de frutanos por planta, enquanto nas plantas controle, que não receberam N suplementar, esse valor foi de 3,5 g. Assim, aplicação de 24 kg N.ha-1 é recomendada para uma colheita antecipada (12 meses) visando à produção de frutanos.

Morfoanatomia comparada dos frutos em desenvolvimento de Vernonia brevifolia Less. e V. herbacea (Vell.) Rusby (Asteraceae)

Neste trabalho, o pericarpo e pápus de Vernonia brevifolia e V. herbacea são descritos morfoanatômica e ontogeneticamente. As espécies são muito similares entre si, possuindo ovário ínfero, bicarpelar, sincárpico e unilocular. Na formação do pericarpo, nenhuma das regiões é multiplicativa. O exocarpo é unisseriado e recoberto por fina cutícula. Observam-se tricomas tectores longos, multicelulares e bisseriados, que persistem até a maturidade; os tricomas glandulares capitados não se mantêm na maturidade. O mesocarpo externo é composto por duas ou três camadas de fibras em V. herbacea e apenas uma em V. brevifolia, acumulando cristais prismáticos. O mesocarpo interno é similar nas espécies, de natureza parenquimática. Feixes vasculares colaterais ocorrem imersos entre o mesocarpo externo e o interno. O endocarpo é unisseriado, havendo duas a três camadas apenas nas regiões de fusão dos carpelos. Na porção apical do pericarpo, observa-se uma protuberância onde se insere o pápus duplo, composto por células lignificadas, algumas projetadas perifericamente. Na base do fruto, ocorre o carpopódio que, em V. herbacea, é mais amplo e abriga drusas e estilóides; em V. brevifolia, o carpopódio é reduzido e não ocorrem cristais. Na maturidade, o pericarpo de ambas as espécies é desidratado, de modo que a maioria das camadas fica colapsada, distinguindo-se apenas algumas células do exocarpo e tricomas tectores, as fibras e cristais mesocárpicos externos e o xilema dos feixes vasculares.

Fructan production in Vernonia herbacea (Vell.) Rusby is related to adequate nitrogen supply and period of cultivation

Previous studies showed that plants of Vernonia herbacea grown for one year under a limited nitrogen supply presented reduced growth and higher fructan content than plants treated with sufficient nitrogen supply. However, the total fructan production was similar in both plant groups due to the higher biomass of the underground reserve organ in nitrogen-sufficient (N-sufficient) plants. In the present study we aimed to evaluate if a stress growing condition under nitrogen-limited (N-limited) supply, following cultivation under N-sufficient supply would have a positive effect on fructan production. Plants cultivated during one year under N-sufficient supply (10.7 mmol L-1 N-NO3-) were separated in two groups. During the following six months, one group continued to receive the same treatment (control) while the other received an N-limited supply (1.3 mmol L-1 N-NO3-). Growth, photosynthesis and soluble carbohydrates were measured at days 0, 30, 60, 90 and 180. At day 30, plants transferred to N-limited supply showed a significant increase in growth and a decrease in fructan concentration, as a response to the stressing condition. However, in the following period growth was reduced and fructan concentration was increased, confirming the inverse relationship between nitrogen concentration and fructan content. After 180 days, although the fructan concentration in N-limited was significantly higher, with a fructan production of 6.0 g plant¹, the higher gain in rhizophore biomass after 18 months of cultivation in N-sufficient solution led to a fructan production of 8.3 g plant¹, thus surpassing the higher fructan concentration of N-limited plants.

Immunization by subcutaneous implants of polyester-polyurethane sponges coupled with antigen

A new protocol is described for immunization of outbred Swiss mice. The procedure is based on subcutaneous implantation of antigen-coupled polyester-polyurethane sponges cut into disks of 10 mm in diameter vs 2 mm in thickness. Antigen coupling was performed by overnight incubation of the sponge with a solution of ovalbumin (Ova) (2 mg/ml) diluted in sodium carbonate buffer, pH 9.6. The amount of ovalbumin that was taken up by the sponge was between 71.4 to 82.5 µg. This was estimated by comparing the Ova absorbance at 280 nm in coating buffer solutions before and after incubation. To compare the efficiency of the proposed method, experimental groups immunized with the antigen in the presence of adjuvants (10 µg in Al(OH)3 or 100 µg in complete Freund's adjuvant (CFA)) were run in parallel. The data obtained after the 3rd week of immunization indicate that both cellular and humoral immune responses were achieved. These were assayed by antigen-induced footpad swelling and ELISA (specific antibodies), respectively. The levels of both immune responses elicited were similar to the responses observed in mice immunized with ovalbumin in the presence of Al(OH)3. The method might represent an advantage when immunizing with pathogenic antigens. Preliminary experiments have suggested that the antigen remains immobilized or bound to the sponge for a long period of time, since there is an increment on the cell population inside the sponges after boosting the animals. If so, the undesirable effects of immunization would be reduced.