In vitro and in vivo genotoxicity and cytotoxicity analysis of protein extract from Aplysina fulva sponges

This study evaluated the physicochemical and morphological properties of a marine sponge protein extract (PE) using scanning electron microscopy (SEM), Energy dispersive X-ray spectroscopy (EDS), analysis of mass loss and pH and in vitro and in vivo. Scanning electron microscopy showed that PE fibers present a granular aspect and irregular structure and the element carbon followed by oxygen was detected in the EDS analysis. Moreover, a 29% of mass loss was observed after 14 days and the pH slightly modified after 14 days. Cell viability of fibroblast cells (L929) of control and PE at a concentration of 25% demonstrated higher values compared to the groups. Osteoblast cell viability of PE at 25 and 50% was significantly higher. Comet assay on day 1 showed higher values for PE at 25%. In addition, in vivo experiments demonstrated that in the treated animals, the bone defects were filled with biomaterial particles, granulation tissue and some areas of newly formed bone. Furthermore, similar immunoexpression of Runx-2 and Cox-2 was observed. Taken together, all results suggest that PE is biocompatible, present non-citotoxicity in the in vitro studies (at the lower concentration) and in the in vivo studies and it can be considered as an alternative source of collagen for tissue engineering proposals.

Biological activities of marine invertebrates extracts from the northeast brazilian coast / Atividades biológicas de extratos de invertebrados marinhos da costa nordeste do Brasil

Abstract This paper reports the in vitro antiproliferative effects, antiprotozoal, anti-herpes and antimicrobial activities of 32 organic extracts of 14 marine sponges and 14 corals collected in northeast Brazilian coast. The ethanolic extracts of the sponges Amphimedon compressa and Tedania ignis, and the acetone extract of Dysidea sp. showed relevant results concerning the antiproliferative effects against A549, HCT-8, and PC-3 cell lines by sulforhodamine B assay, but also low specificity. Concerning the antiprotozoal screening, the ethanolic extract of Amphimedon compressa and the acetone and ethanolic extracts of Dysidea sp. were the most active against Leishmania amazonensis and Trypanosoma cruzi expressing β-galactosidase in THP-1 cells. In the preliminary anti-HSV-1 (KOS strain) screening, the ethanolic extracts of the sponges Amphimedon compressa, Haliclona sp. and Chondrosia collectrix inhibited viral replication by more than 50%. The most promising anti-herpes results were observed for the ethanolic extract of Haliclona sp. showing high selective indices against HSV-1, KOS and 29R strains (SI> 50 and >79, respectively), and HSV-2, 333 strain (IS>108). The results of the antibacterial screening indicated that only the ethanolic extract of Amphimedon compressa exhibited a weak activity against Enterococcus faecalis, Pseudomonas aeruginosa and Escherichia coli by the disk diffusion method. In view of these results, the extracts of Amphimedon compressa, Tedania ignis and Dysidea sp. were selected for further studies aiming the isolation and identification of the bioactive compounds with antiproliferative and/or antiprotozoal activities. The relevant anti-herpes activity of the ethanolic extract of Haliclona sp. also deserves special attention, and will be further investigated.

Resumo Este artigo reporta as atividades in vitro antiproliferativa, atiprotozoárica, anti-herpética e antimicrobiana de 32 extratos orgânicos provenientes de 14 esponjas marinhas e 14 corais coletados no litoral nordestino brasileiro. Os extratos etanólicos das esponjas Amphimedon compressa e Tedania ignis, e o extrato acetônico de Dysidea sp. demonstraram resultados promissores em relação aos efeitos antiproliferativos frente as linhagens celulares A549, HCT-8, PC-3 pelo método da sulforrodamina B, mas sem especificidade. Em relação à atividade antiprotozárica, os extratos etanólico de Amphimedon compressa e acetônico e etanólico de Dysidea sp. apresentaram atividade contra Leishmania amazonensis e Trypanosoma cruzi através do método de expressão de β-galactosidase em células THP-1. Na investigação preliminar de atividade antiviral frente ao vírus Herpes simplex tipo 1 (cepa KOS), os extratos etanólicos das esponjas Amphimedon compressa, Haliclona sp. e Chondrosia collectrix inibiram mais de 50% da replicação viral. O extrato etanólico da esponja Haliclona sp. demonstrou resultados promissores para atividade anti-herpética com altos índices de seletividade para as cepas KOS (IS >50) e 29R (IS>79) frente ao VHS-1 e cepa 333 (IS>108) frente ao VHS-2. O extrato etanólico da esponja Amphimedon compressa exibiu uma pequena atividade contra Enterococcus faecalis, Pseudomonas aeruginosa and Escherichia coli pelo método de difusão em disco. De acordo com os resultados apresentados, os extratos das esponjas Amphimedon compressa, Tedania ignis e Dysidea sp. serão selecionados para futuros estudos de isolamento e identificação dos compostos bioativos para as atividades antiproliferativa e antiprozoárica. O extrato etanólico de Haliclona sp. será investigado por possuir atividade relevante anti-herpética.

In Vivo Biological Effects of Marine Biosilica on a Tibial Bone Defect in Rats

Abstract Research on biomaterials of natural origin has gained prominence in the literature. Above all, marine sponges, due to their architecture and structural components, present a promising potential for the engineering of bone tissue. In vitro studies demonstrate that a biosilica of marine sponges has osteogenic potential. However, in vivo works are needed to elucidate the interaction of biosilica (BS) and bone tissue. The objective of the study was to evaluate the morphological and chemical characteristics of BS compared to Bioglass (BG) by scanning electron microscopy (SEM) and X-ray dispersive energy (EDX) spectroscopy. In addition, to evaluate the biological effects of BS, through an experimental model of tibial bone defect using histopathological, histomorphometric, immunohistochemical (IHC) and mechanical tests. SEM and EDX demonstrated the successful extraction of BS. Histopathological analysis demonstrated that Control Group (GC) had greater formation of newly formed bone tissue compared to BG and BS, yet BG bone neoformation was greater than BS. However, BS showed material degradation and granulation tissue formation, with absence of inflammatory process and formation of fibrotic capsule. The results of histomorphometry corroborate with those of histopathology, where it is worth emphasizing the positive influence of BS in osteoblastic activity. IHQ demonstrated positive VEGF and TGF-β immunoexpression for GC, BS and BG. In the mechanical test no significant differences were found. The present results demonstrate the potential of BS in bone repair, further studies are needed other forms of presentation of BS are needed.

Cold-active pectinolytic activity produced by filamentous fungi associated with Antarctic marine sponges

BACKGROUND: Pectinase enzymes catalyze the breakdown of pectin, a key component of the plant cell wall. At industrial level, pectinases are used in diverse applications, especially in food-processing industry. Currently, most of the industrial pectinases have optimal activity at mesophilic temperatures. On the contrary, very little is known about the pectinolytic activities from organisms from cold climates such as Antarctica. In this work, 27 filamentous fungi isolated from marine sponges collected in King George Island, Antarctica, were screened as new source of cold-active pectinases. RESULTS: In semi-quantitative plate assays, 8 out 27 of these isolates showed pectinolytic activities at 15 °C and one of them, Geomyces sp. strain F09-T3-2, showed the highest production of pectinases in liquid medium containing pectin as sole carbon source. More interesting, Geomyces sp. F09-T3-2 showed optimal pectinolytic activity at 30 °C, 10 °C under the temperature of currently available commercial mesophilic pectinases. CONCLUSION: Filamentous fungi associated with Antarctic marine sponges are a promising source of pectinolytic activity. In particular, pectinases from Geomyces sp. F09-T3-2 may be potentially suitable for biotechnological applications needing cold-active pectinases. To the best of our knowledge, this is the first report describing the production of pectinolytic activity from filamentous fungi from any environment in Antarctica.

Ecology and biotechnological potential of bacterial community from three marine sponges of the coast of Rio de Janeiro, Brazil

ABSTRACT Marine sponges has been a large reservoir of microbial diversity, with the presence of many species specific populations as well as producing biologically active compounds, which has attracted great biotechnological interest. In order to verify the influence of the environment in the composition of the bacterial community present in marine sponges and biotechnological potential of bacteria isolated from these organisms, three species of sponges and the waters surrounding them were collected in different beaches of Rio de Janeiro, Brazil. The profile of the bacterial community present in sponges and water was obtained by PCR-DGGE technique and the biotechnological potential of the strains isolated by producing amylase, cellulase, protease and biosurfactants. The results showed that despite the influence of the environment in the composition of the microbial community, studied marine sponges shown to have specific bacterial populations, with some, showing potential in the production of substances of biotechnological applications.

Antibiotic resistance genes detected in the marine sponge Petromica citrina from Brazilian coast

ABSTRACT Although antibiotic-resistant pathogens pose a significant threat to human health, the environmental reservoirs of the resistance determinants are still poorly understood. This study reports the detection of resistance genes (ermB, mecA, mupA, qnrA, qnrB and tetL) to antibiotics among certain culturable and unculturable bacteria associated with the marine sponge Petromica citrina. The antimicrobial activities elicited by P. citrina and its associated bacteria are also described. The results indicate that the marine environment could play an important role in the development of antibiotic resistance and the dissemination of resistance genes among bacteria.

Investigation of freshwater sponges spicules deposits in a karstic lake in Brazil / Investigação de depósito de espículas de esponjas de água doce em um lago cárstico no Brasil

Abstract The environmental conditions which contributed to the formation of the notorious quaternary deposits of freshwater sponge spicules in karstic lentic environments in Brazil have been subject of some speculation. No investigation has yet been conducted to test whether these deposits currently originate in karstic lakes. To provide for such an investigation, Serra Negra Lake, which is formed on an ultramafic-alkaline-carbonatite dome at central western Brazil, close to the area of occurrence of the paleo-deposits was selected for the study. Bottom sediments were sampled at 10 stations across the lake, and water was sampled at five of the stations, in June/2011 (rainy season) and October/2011 (dry season). Analysis of granulometry, organic matter and presence of spicules were carried out in the sediments. Lake water was analysed for the main physical and chemical characteristics. Deposit of spicules was restricted to the northern area of the lake, which is rich in macrophyte. The taxonomic analysis of the spicules indicated the contribution of five sponge species, Dosilia pydanieli, Metania spinata, Radiospongilla amazonensis, Trochospongilla variabilis and Heterorotula fistula, which formed large deposits in neighbouring areas. The high silica concentration, derived from the dome volcanic rocks, constant water level and available substrate are credited for the continuous production of sponges and spicules, confirmed by the rare presence of gemmoscleres. The lake is classed as a minerotrophic fen type of bog with a heavy contribution from the surrounding creeks. Lake sediments are fine with high levels of organic matter and peat, which contributed to the trapping of spicules in the sediments.

Resumo As condições ambientais que contribuíram para a formação de notórios depósitos quaternários de espículas de esponjas de água doce em ambientes cársticos no Brasil têm sido objeto de algumas especulações. Nenhuma investigação foi ainda realizada para testar se esses depósitos originam-se atualmente em lagos cársticos. Para prover tal investigação, a Lagoa Serra Negra, formada em um domo ultramáfico-alcalino-carbonatítico, no centro oeste do Brasil, próximo à área de ocorrência dos referidos paleo-depósitos, foi selecionada para o estudo. Sedimentos de fundo foram amostrados em 10 estações ao longo do lago, e a água foi coletada em cinco das estações, em junho/2011 (estação chuvosa) e outubro/2011 (estação seca). Análises de granulometria, matéria orgânica e presença de espículas foram realizadas nos sedimentos. A água do lago foi analisada para as principais características físicas e químicas. O depósito de espículas foi restrito à área norte do lago, que é rico em macrófitas. A análise taxonômica das espículas indicou a contribuição de cinco espécies de esponjas, Dosilia pydanieli, Metania spinata, Radiospongilla amazonensis, Trochospongilla variabilis e Heterorotula fistula, que formaram grandes depósitos em áreas vizinhas. A alta concentração de sílica, derivada das rochas vulcânicas do domo, o nível constante da água e a disponibilidade de substrato são creditadas à produção contínua de esponjas e espículas, confirmada pela presença rara de gemoscleras. O lago é classificado como uma turfeira do tipo fen, minerotrófica, com forte contribuição dos arroios circundantes. Os sedimentos da lagoa são finos, com altos níveis de matéria orgânica e formação de turfa, o que contribuiu para a captura das espículas nos sedimentos.

Middle ear packing materials: comparison between absorbable hemostatic gelatine sponge and sugarcane biopolymer sponge in rats / Materiais de suporte na orelha média: comparação entre a esponja de gelatina absorvível e a esponja do biopolímero da cana-de-açúcar em ratos

INTRODUCTION: Several biomaterials can be used in ear surgery to pack the middle ear or support the graft. The absorbable gelatin sponge is the most widely used, but it may produce fibrosis and impair ventilation of the middle ear. OBJECTIVE: This experimental study aimed to investigate the inflammatory effects of the sugarcane biopolymer sponge (BP) in the rat middle ear compared with absorbable gelatin sponge (AGS). MATERIALS AND METHODS: Prospective experimental study design. Thirty adult female Wistar rats were allocated to receive the BP sponge into the right ear and AGS into the left ear. Animals were randomly killed at 4 and 12 weeks post-procedure. Qualitative histological assessments were performed to evaluate the inflammatory reaction in the tympanic bullae. RESULTS: The BP sponge caused inflammation more intense and persistent than AGS. The BP was not absorbed during the experiment. Fibrosis was observed only in the ears with AGS. There were thickening of the mucosa and neoangiogenesis in the group of AGS. CONCLUSION: Despite inflammation, the BP sponge produced less fibrosis and neoangiogenesis compared to AGS. The sponge BP appeared to be a non-absorbable biomaterial in the middle ear. .

INTRODUÇÃO: Existem diversos biomateriais que podem ser utilizados na cirurgia otológica para preencher a cavidade da orelha média ou dar suporte a enxertos. A esponja de gelatina absorvível é a mais utilizada, mas pode provocar fibrose e prejudicar a ventilação da orelha média. OBJETIVO: Investigar os efeitos da reação inflamatória provocada pela esponja do biopolímero da cana-de-açúcar (BP) comparada a esponja de gelatina absorvível (EGA) na mucosa da orelha média de ratos. MATERIAIS E MÉTODOS: Estudo experimental prospectivo. A esponja do BP foi implantada na orelha direita e a EGA na orelha esquerda de 30 ratos Wistar fêmeas. Os animais foram sacrificados com 4 e 12 semanas após o procedimento. Avaliação histológica qualitativa foi realizada para verificar a reação inflamatória na bula timpânica. RESULTADOS: A esponja do BP provocou exsudato inflamatório mais intenso e persistente que a EGA. O BP não foi absorvido durante o tempo de observação. Traves de fibrose foram observadas apenas nos ouvidos com a EGA. Houve espessamento da mucosa e neoangiogênese no grupo da EGA. CONCLUSÃO: Apesar da reação inflamatória, a esponja do BP provocou menos fibrose e neoangiogênese quando comparada a EGA. A esponja do BP comportou-se como um biomaterial não absorvível na orelha média. .

Efectos de extractos de Neopetrosia rosariensis sobre mieloperoxidasa y elastasa con la utilización del modelo neutrófilos / Effects of extracts from Neopetrosia rosariensis on myeloperoxidase and on elastase by using the neutrophil model

Introducción: los neutrófilos son la primera línea de defensa, los cuales liberan su contenido granular, constituido entre otros por enzimas generadoras de especies reactivas del oxígeno como mieloperoxidasa y enzimas proteolíticas como la elastasa, que degradan elastina y colágeno. Objetivo: evaluar la toxicidad de los extractos de la esponja marina Neopetrosia rosariensis y su acción como inhibidores de la desgranulación del neutrófilo, e inhibición de la actividad catalítica de la elastasa de neutrófilos humanos. Métodos: se determinó la toxicidad por el método de exclusión del colorante azul de tripano. Se evaluó la desgranulación por la disminución del porcentaje de liberación de la mieloperoxidasa, y se determinó la inhibición de la actividad catalítica de la elastasa de neutrófilos humanos por la reducción de la actividad de la enzima de un sustrato específico. Resultados: los extractos presentaron baja toxicidad a las concentraciones evaluadas, con porcentaje de viabilidad celular superior al 80 por ciento, incluso a la concentración más alta utilizada (25 µg/mL). También presentaron actividad inhibitoria de la liberación de mieloperoxidasa, concentrándose el efecto en los extractos más polares (metanólico total y parcial) y la inhibición de la actividad de la elastasa en los de más baja polaridad (hexano y diclorometano parcial), a concentraciones de 10 y 25 µg/mL, acorde con la naturaleza lipofílica del sitio activo de la enzima. Conclusiones: los resultados en los extractos evaluados son promisorios, teniendo en cuenta que cada muestra representa una mezcla compleja de compuestos activos entre 10 y 25 µg/mL, lo que resulta de mucho interés para continuar con su fraccionamiento como agentes potenciales para la posible terapia de enfermedades que presentan procesos inflamatorios(AU)

Introduction: neutrophils are the first line of defense, releasing their granular contents made up of reactive oxygen species-generating enzymes such as myeloperoxidase and proteolytic enzymes such as elastase, which degrade elastin and collagen. Objective: to evaluate toxicity of marine sponge Neopetrosia rosariensis extracts and their action as inhibitors of neutrophil degranulation and inhibition of the catalytic activity of human neutrophil elastase. Method: the cytotoxicity was assessed by Trypan blue exclusion test of cell viability. Degranulation inhibition was evaluated by reduction of myeloperoxidase release percentage and inhibition of catalytic activity of elastase was determined by reduction of enzyme activity on specific substrate. Results: the extracts exhibited low toxicity at the evaluated concentrations, with cellular viability percentage exceeding 80 percent even at the highest concentration (25 µg/mL). They also showed inhibitory action of myeloperoxidase release, the greatest effect being concentrated on the most polar extracts (total and partial methanolic). The inhibition of the elastase activity was mostly found in the lowest polarity extracts (hexane and partial dichloromethane) at concentrations of 10 and 25 µg/mL, according to the hydrophobic character of the active site of the enzyme. Conclusions: these results of the evaluated extracts are encouraging, considering each extract represents a complex mixture of active compounds of 10-25 µg/mL. This arouses great interest to continue their fractioning as potential agents for possible therapy of inflammatory processes(AU)

Acidobacteria appear to dominate the microbiome of two sympatric Caribbean Sponges and one Zoanthid

BACKGROUND: Marine invertebrate-associated microbial communities are interesting examples of complex symbiotic systems and are a potential source of biotechnological products. RESULTS: In this work, pyrosequencing-based assessment from bacterial community structures of sediments, two sponges, and one zoanthid collected in the Mexican Caribbean was performed. The results suggest that the bacterial diversity at the species level is higher in the sediments than in the animal samples. Analysis of bacterial communities' structure showed that about two thirds of the bacterial diversity in all the samples belongs to the phyla Acidobacteria and Proteobacteria. The genus Acidobacteriumappears to dominate the bacterial community in all the samples, reaching almost 80% in the sponge Hyrtios. CONCLUSIONS: Our evidence suggests that the sympatric location of these benthonic species may lead to common bacterial structure features among their bacterial communities. The results may serve as a first insight to formulate hypotheses that lead to more extensive studies of sessile marine organisms' microbiomes from the Mexican Caribbean.

Bromotyrosine derivatives from marine sponges inhibit the HIV-1 replication in vitro / Replication in vitro bromotirosinas derivadas de esponjas marinas inhiben la replicación in vitro del VIH-1

Background: Human immunodeficiency virus type 1 (HIV-1) infection and Acquired immunodeficiency syndrome are mayor global public health issues. HIV-1 infection is now manageable as a chronic disease thanks to the development of antiretroviral therapy; however, the existence of HIV drug resistance and collateral effects have increased the search for therapeutic alternatives. Compounds of marine resources have been studied for their antiviral potential. Objectives: To evaluate the antiviral activity of isolated bromotyrosine-derivative compounds from the Colombian marine sponges, Verongula rigida and Aiolochoria crassa against HIV-1 infection in vitro. Methods: Cytotoxicity of 11 bromotyrosine-derivative compounds was determined by the MTT assay. Inhibition of HIV-1 replication was performed using the U373-MAGI cell line, which was infected with recombinant green fluorescent protein (GFP)-expressing viruses pseudotyped, in the presence or absence of the compounds. The percentage of infected cells was evaluated by flow cytometry. In addition, the inhibition of reverse transcription and nuclear import was determined by quantification of early and late reverse transcription products and 2-LTR circles, respectively, using quantitative PCR. Results: Aeroplysinin-1, purealidin B and 3-bromo-5-hydroxy-Omethyltyrosine inhibited the HIV-1 replication in a dose-dependent manner, with a median maximum percentage of inhibition of 74% (20 µM), 57% (80 µM) and 47% (80 µM), respectively. Importantly, none of these concentrations were cytotoxic. Aeroplysinin-1, 19-deoxyfistularin 3, purealidin B, fistularin 3 and 3-bromo-5-hydroxy-O-methyltyrosine inhibited the nuclear import efficiently; while 3,5-dibromoN,N,N,O-tetramethyltyraminium, aeroplysinin-1, purealidin B, fistularin 3 and 3-bromo-5-hydroxy-Omethyltyrosine inhibited X4 HIV-1 cell entry with a median maximum percentage of inhibition ranging between 2 to 30%. Conclusions: Aeroplysinin-1, 19-deoxyfistularin 3, purealidin B, fistularin 3 and 3-bromo-5-hydroxy-O-methyltyrosine inhibited HIV replication at different steps. This study opens the possibility of chemically synthesizing these compounds and evaluating them as alternative therapies against HIV-1.

Estructura de la comunidad de esponjas (Porifera) en tres arrecifes del Parque Nacional Morrocoy, Venezuela y su relación con algunas variables ambientales / Community structure of sponges (Porifera) in three reefs at Morrocoy National Park, Venezuela and its correspondence with some environmental variables

Sponges have an important ecological role in coral reef ecosystems. However, when compared to other benthic Phyla, it has been little researched. This research was focused in the variability of the community structure of sponges in three locations at Morrocoy National Park (Cayo Sombrero, Playa Mero and Punta Brava) exposed to different environmental conditions (transparency and currents intensity) and affected in different degree of severity by a mass mortality event in 1996. A total of 15 transects (10m long and 1m wide) were evaluated in three strata (between 3 and 15m depth) in each site, where all the individuals were counted by species. Relative abundance by species, diversity and evenness were calculated. Locations showed differences respect turbidity, wave and current intensity. 27 species were found in Morrocoy; Cayo Sombrero (23), Playa Mero (18) and Punta Brava (15). Agelas sceptrum, Amphimedon erina and Niphates erecta were the most common in first location; Niphates erecta and Dysidea etheria in Playa Mero and Dysidea etheria, Niphates erecta and Amphimedon erina in Punta Brava. The species composition showed statistical differences between all three locations; Cayo Sombrero resulted the most diverse and even, followed by Playa Mero and Punta Brava. According to Sorensen Similarity Index results, Cayo Sombrero and Playa Mero were more similar, while Punta Brava resulted the most different. The variability in environmental conditions and the differential mass mortality effects of 1996 in all three reefs, were probably the main causes of the differences between their sponge communities. Nevertheless, we cannot conclude about the weight of these factors.

El presente estudio pone en evidencia la variación en la estructura de la comunidad de esponjas en tres localidades del Parque Nacional Morrocoy (Cayo Sombrero, Playa Mero y Punta Brava) con distintas condiciones ambientales y afectadas diferencialmente por una mortalidad masiva ocurrida en 1996. En un total de 15 transectos de 10m de largo y 1m de ancho en cuatro estratos comprendidos entre 3 y 15m de profundidad en cada localidad; se contabilizaron los individuos por especie para calcular la abundancia relativa, diversidad y equidad. Se analizaron las variables ambientales por localidad y se aprecian diferencias entre ellas con respecto a la corriente, turbidez, y exposición al oleaje. Fueron totalizadas 27 especies; Cayo Sombrero (23), Playa Mero (18) y Punta Brava (15), en la primera localidad domina: Agelas sceptrum, Amphimedon erina y Niphates erecta, en Playa Mero: Niphates erecta y Dysidea etherea y en Punta Brava: Dysidea etherea, Niphates erecta y Amphimedon erina. La composición de especies mostró diferencias estadísticamente significativas entre localidades. La mayor diversidad y equidad correspondió a Cayo Sombrero; donde las condiciones fueron más favorables para el crecimiento de estos organismos, seguido de Playa Mero y Punta Brava, guardando las dos primeras mayor similitud entre sí, según Índice de Sorensen.

Antifouling activity of twelve demosponges from Brazil / Atividade anti-incrustante de doze demosponjas do Brasil

Benthic marine organisms are constantly exposed to fouling, which is harmful to most host species. Thus, the production of secondary metabolites containing antifouling properties is an important ecological advantage for sessile organisms and may also provide leading compounds for the development of antifouling paints. High antifouling potential of sponges has been demonstrated in the Indian and Pacific oceans and in the Caribbean and Mediterranean seas. Brazilian sponges remain understudied concerning antifouling activities. Only two scientific articles reported this activity in sponges of Brazil. The objective of this study was to test crude extracts of twelve species of sponges from Brazil against the attachment of the mussel Perna perna through laboratorial assays, and highlight promising species for future studies. The species Petromica citrina, Amphimedon viridis, Desmapsamma anchorata, Chondrosia sp., Polymastia janeirensis, Tedania ignis, Aplysina fulva, Mycale angulosa, Hymeniacidon heliophila, Dysidea etheria, Tethya rubra, and Tethya maza were frozen and freeze-dried before extraction with acetone or dichloromethane. The crude extract of four species significantly inhibited the attachment of byssus: Tethya rubra (p = 0.0009), Tethya maza (p = 0.0039), Petromica citrina (p = 0.0277), and Hymeniacidon heliophila (p = 0.00003). These species, specially, should be the target of future studies to detail the substances involved in the ability antifouling well as to define its amplitude of action.

Organismos bentônicos marinhos estão expostos constantemente à incrustação, que pode ser danosa para a maioria das espécies. Assim, a produção de metabólitos secundários com propriedades anti-incrustantes é uma vantagem ecológica importante para organismos sésseis e pode também orientar o estudo de substâncias para o desenvolvimento de tintas anti-incrustantes. O alto potencial anti-incrustante de esponjas tem sido demonstrado nos oceanos Índico e Pacífico, nos mares Mediterrâneo e Caribenho. Esponjas brasileiras permanecem pouco estudadas em relação à atividade anti-incrustante. Apenas dois artigos científicos registraram essa atividade em esponjas do Brasil. O objetivo desse estudo foi testar os extratos brutos de doze espécies de esponjas do Brasil contra a fixação do molusco Perna perna através de ensaios laboratoriais e também destacar espécies promissoras para estudos futuros. As espécies Petromica citrina, Amphimedon viridis, Desmapsamma anchorata, Chondrosia sp., Polymastia janeirensis, Tedania ignis, Aplysina fulva, Mycale angulosa, Hymeniacidon heliophila, Dysidea etheria, Tethya rubra e Tethya maza foram congeladas e liofilizadas logo após a coleta e posteriormente procedeu-se a extração com acetona ou diclorometano. O extrato bruto de quatro espécies inibiu significativamente a fixação de bissos: Tethya rubra (p = 0.0009), Tethya maza (p = 0.0039), Petromica citrina (p = 0.0277), e Hymeniacidon heliophila (p = 0.00003). Essas espécies, especialmente, devem ser priorizadas em estudos futuros para detalhamento das substâncias envolvidas na capacidade anti-incrustante, bem como para definir sua amplitude de ação.

Evaluation of antimicrobial activity and cell viability of Aloe vera sponges

Background: Aloe vera L., member of the Liliaceae family, has been shown to stimulate cell proliferation and contribute to healing and angiogenesis, has anti-bacterial, anti-fungal and anti-inflammatory activity. In addition, Aloe vera can be used as a support for drug transport. Our objective is to evaluate antimicrobial activity and cytotoxicity of sponges of Aloe vera L. for use as a carrying support of drugs. Results: In this work, sponge of free Aloe vera (AV) loaded with amoxicillin (AMX) or nystatin (NYS) at 1% w/w, were prepared and physico-chemically characterized via X-ray diffraction, Fourier Transform Infrared Spectroscopy and thermal analysis. Antimicrobial potency of AV sponge alone, loaded with AMX or NYS, against strains of Streptococcus mutans, Staphylococcus aureus, Aggregatibacter actinomycetemcomitans, Enterococcus faecalis and Candida albicans was determined. Osteoblasts and human gingival fibroblasts were cultivated on AV, Aloe vera loaded with amoxicillin (AV/AMX) and Aloe vera loaded with nystatin (AV/NYS) and cellular viability was assessed. The physico-chemical characterization performed suggested that the loaded drugs were dispersed in the sponge and those interactions between the AV sponge and the loaded drugs were weak. Furthermore, AV loaded with AMX or NYS demonstrated antimicrobial potency and osteoblasts and fibroblasts were viable after 24 hrs on free AV, and AV loaded with AMX or NYS. Conclusions: Our results indicate that sponges of free AV, loaded with AMX or NYS, are biocompatible and exhibit antimicrobial activity.

Diversity of the candidate phylum Poribacteria in the marine sponge Aplysina fulva

Poribacterial clone libraries constructed for Aplysina fulva sponge specimens were analysed with respect to diversity and phylogeny. Results imply the coexistence of several, prevalently "intraspecific" poribacterial genotypes in a single sponge host, and suggest quantitative analysis as a desirable approach in studies of the diversity and distribution of poribacterial cohorts in marine sponges

Actividad antiplasmodial in vivo de las esponjas Mycale laxissima y Clathria echinata / In vivo antiplasmodial activity of Mycale laxissima and Clathria echinata sponges

Introducción: la búsqueda de nuevas sustancias antimaláricas incluye entre sus retos el desarrollo de alternativas para el tratamiento de la malaria cerebral, debido a la alta mortalidad y las deficiencias neurológicas persistentes después del tratamiento con los medicamentos recomendados actualmente. Objetivos: evaluar la actividad de fracciones orgánicas de Mycale laxissima y Clathria echinata en el modelo de malaria cerebral producida por la infección de ratones C57BL/6 con Plasmodium berghei ANKA. Métodos: se evaluaron fracciones orgánicas obtenidas mediante cromatografía flash en fase inversa a partir de los extractos crudos de las dos especies. Se realizó un análisis químico cualitativo para detectar la presencia de saponinas, triterpenos/esteroides y alcaloides en estas fracciones. El efecto esquizonticida de las fracciones se determinó mediante la prueba de supresión de la parasitemia al inicio de la infección. Se evaluaron la supervivencia, los síntomas neurológicos y la reducción del peso corporal en los días subsiguientes. Resultados: las fracciones orgánicas de Mycale laxissima a dosis de 200 mg/kg y Clathria echinata a100 mg/kg no mostraron una disminución sustancial del peso de los animales o muertes hasta el día 4; para las cuales se obtuvieron reducciones significativas de las medianas de la parasitemia de 45 % y 53 %, respectivamente. La fracción de Mycale laxissima a 200 mg/kg produjo un incremento significativo en el tiempo de supervivencia hasta 20 d, mientras los animales tratados con Clathria echinata a 100 mg/kg presentaron una mediana de tiempo de supervivencia de 16 d. Ambos incrementos fueron superiores a 7 d. En este período, los animales tratados con las fracciones orgánicas de Clathria echinata y Mycale laxissima no presentaron los síntomas neurológicos observados en los controles. Esta prolongación de la supervivencia fue similar a la observada en presencia de dosis de 7,5 mg/kg de cloroquina. Conclusiones: las fracciones orgánicas de Mycale laxissima y Clathria echinata mostraron actividades antimaláricas, promisorias en el modelo de infección de ratones C57BL/6 con Plasmodium berghei ANKA, que sugieren el estudio de sus constituyentes químicos activos.

Introduction: the search of new antimalarial compounds comprises, among its challenges, the development of therapeutic alternatives for cerebral malaria; due to the high mortality and neurological deficiencies that persist after treatment with recommended drugs. Objectives: to evaluate the activity of organic fractions of Mycale laxissima and Clathria echinata in the cerebral malaria model of infection of C57BL/6 mice with Plasmodium berghei ANKA. Methods: preparative fractions of both species were obtained by reverse-phase flash chromatography. In order to detect the presence of saponins, triterpenods/steroids and alkaloids, a qualitative chemical analysis was performed. The schyzontocidal effect of the extracts was determined by the suppression test at the beginning of the infection. Survival, neurological symptoms and body weight changes were evaluated in subsequent days. Results: the organic fractions of Mycale laxissima at 200 mg/kg and Clathria echinata at 100 mg/kg showed neither substantial reductions of body weights, nor deaths of animals until day 4; but caused significant reductions of median parasitemia of 45 % and 53 % respectively. The fraction of Mycale laxissima at 200 mg/kg caused a significant increase in the median survival time up to day 20, whereas animals treated with Clathria echinata at 100 mg/kg presented a survival of 16 days. Both increases the survival time 7 days. Neurological alterations were not observed in the groups treated with organic fractions when compared to the control group. This survival extension was similar to the effect of administration of 7.5 mg/kg of chloroquine. Conclusions: the organic fractions of Mycale laxissima and Clathria echinata exhibited promising antimalarial activities in the infection model of C57BL/6 mice with Plasmodium berghei ANKA. This indicates that their active chemical constituents should be studied.

Isolation and cultivation of fungal strains from in vitro cell cultures of two marine sponges (Porifera: Halichondrida and Haplosclerida)

Despite the large number of reports describing sponge-microbe associations, limited knowledge is available about associated fungi and their relationships with the hosts. In this work, specific fungal strains were obtained directly from in vitro sponge cell cultures (primmorphs) and single sponge cells (cytospins) and compared with those obtained from whole tissue preparations. A total of 27 fungal strains were isolated from the marine sponges Hymeniacidon heliophila and Haliclona melana. Fifteen strains, nine from H. heliophila and six from H. melana, were obtained from whole tissue and were considered as possible mesohyl associated or transient fungi. Twelve strains were isolated from in vitro sponge cell cultures (primmorphs) and were, therefore, considered as cell associated. From these, five different strains were obtained from H. heliophila isolated cells, while five were identified from cytospins and two from primmorphs of H. melana. The fungal strains obtained from cell cultures from both sponge species were different, and none of them were detected in the whole tissue preparations of the same species. Nine H. heliophila and seven H. melana strains shows low similarity with the sequences available in public databases and belong to potentially new species. This is the first report of fungi isolated directly from sponge cells, which allowed the observation and selection of specific strains that probably would not be obtained by usual culture dependent techniques.

Environmental state and tendencies of the Puerto Morelos CARICOMP site, Mexico

The CARICOMP site at Puerto Morelos, Mexico was monitored from 1993 to 2005. No significant changes in air temperature, wind patterns, periodicity and quantity of rainfall, sea-surface temperature and water transparency were observed between sampling years. During the study four hurricane impacts were registered. At the coral reef site overall mean cover of fleshy algae (47%) and turf algae (36%) were high, whereas cover of corals (2%) and sponges (3%), and abundance of sea-urchins (0.04 org m-2) were consistently low. Gorgonians were dominant and showed changes in their community structure; the number of species increased from 1993 to 1995, their abundance decreased after Hurricane Roxanne (1995) and recovered by 2001. At four seagrass sites total community biomass remained constant (707.1-929.6 g dry m-2) but the above-ground biomass of the seagrass Syringodium filiforme and fleshy algae increased gradually. Total biomass (531-699 g dry m-2) and leaf productivity (0.89-1.56 g dry m-2 d-1) of the seagrass Thalassia testudinum remained constant, but the species invested proportionally more biomass in above-ground leaf tissues at the end of the study. The minor hurricanes from 1993 until 2005 had no detectable impacts on the seagrass beds, however, the major Hurricane Wilma (October 2005) changed the community composition at three stations and caused complete burial of the vegetation at a coastal station. The gradual changes in the seagrass and reef communities recorded in the 12 years of continuous monitoring of the CARICOMP site may reflect the increased pollution caused by the rapid augment in urban and tourist developments along the coasts and inland from Puerto Morelos, coupled with poor water management practices. Rev. Biol. Trop. 58 (Suppl. 3): 23-43. Epub 2010 October 01.

El sitio CARICOMP en Puerto Morelos, México, se monitoreo de 1993 a 2005. No se registraron cambios significativos en la temperatura del aire, los patrones de viento, la periodicidad y cantidad de lluvia, la temperatura superficial del mar y la transparencia del agua entre los años de muestreo. Durante el estudio se registraron cuatro huracanes menores. En el sitio del arrecife coralino la cobertura global promedio de algas carnosas (47%) y de algas tipo tapete (36%) fue elevada, mientras que la cobertura de corales (2%), esponjas (3%) y la abundancia de erizos (0.04 m-2) fue consistentemente baja. Los gorgonáceos fueron dominantes y mostraron cambios en su estructura comunitaria; el número de especies aumentó de 1993 a 1995, su abundancia disminuyó después del Huracán Roxanne (1995) y se recuperó en el 2001. En cuatro estaciones de pastos marinos la biomasa total de la comunidad permaneció constante (707.1-929.6 gps m-2) pero la biomasa sobre el sustrato del pasto Syringodium filiforme y de algas carnosas aumentó gradualmente. La biomasa total (531-699 gps m-2) y productividad de hojas (0.89-1.56 gps m-2 d-1) del pasto Thalassia testudinum permaneció constante, pero la especie invirtió proporcionalmente más biomasa en el tejido de hojas sobre el sustrato al final del estudio. Los huracanes menores entre 1993 y 2005 no produjeron impactos detectables en las camas de pastos, sin embargo, el huracán mayor Wilma (Octubre 2005) cambió la composición de la comunidad en tres estaciones y causó el enterramiento completo de la vegetación en la estación costera. El cambio gradual en las comunidades de pastos marinos y arrecife registrado en 12 años de monitoreo continuo en el sitio CARICOMP puede reflejar un aumento en la contaminación causada por el rápido incremento en desarrollo urbano y turístico a lo largo de la costa y tierra adentro de Puerto Morelos, acompañado por malas prácticas de manejo del agua.

Distribución de esponjas (Porifera) a lo largo de un gradiente de profundidad en un arrecife coralino, Parque Nacional San Esteban, Carabobo, Venezuela

Sponges (Porifera) distribution along a depth gradient in a coral reef, Parque Nacional San Esteban, Carabobo, Venezuela. Sponges constitute one of the most diverse and abundant animal groups in the marine tropical benthos especially in coral reefs, though poorly studied to species level. The aim of this study is to characterize the sponge community along a depth gradient at Isla Larga (Parque Nacional San Esteban, Venezuela) fringe reef. Net and total sedimentation, roughness index, sponge species richness, density and proportion of the bottom covered by sponges, were evaluated at seven depths (1, 3, 6, 9, 12, 15, 18m), 17 species were identified grouped in 10 demosponges families. The highest densities and coverage corresponded to 6m of depth (6.03ind/m2; 11%), that coincides with the lowest net sedimentation and highest substrate heterogeneity. Most abundant species were Desmapsamma anchorata, Amphimedon erina and Scopalina rueztleri. Principal component analysis divided this community in three zones according to depth. The shallow zone of the reef (1 and 3m), where wave force and high irradiance exert a constant "stress sponges, shows the lowest density and coverage by sponges. In contrast, medium depth (6, 9 y 12m) and deep zone (15 y 18m) with lower light and sedimentation levels seem to enhance sponge growth and survival that are reflected on the higher densities and coverage of sponges. Rev. Biol. Trop. 58 (Suppl. 3): 175-187. Epub 2010 October 01.

Las esponjas son uno de los grupos de animales sésiles más abundantes y diversos de los fondos marinos tropicales, siendo un componente importante en los arrecifes coralinos, aunque poco estudiado a nivel de especies. El objetivo de este estudio fue caracterizar la comunidad de esponjas en el gradiente de profundidad de un arrecife coralino en Isla Larga, Parque Nacional San Esteban, Venezuela. Se trabajaron siete profundidades (1, 3, 6, 9, 12, 15 y 18m) donde se midió la sedimentación neta y bruta, el índice de rugosidad, y se evaluó la riqueza, densidad y cobertura de las esponjas. Se identificaron 17 especies en 10 familias. La mayor densidad y cobertura se encontró a los 6m (6.03ind/m2, 11%), coincidiendo con la mínima sedimentación neta y la máxima rugosidad del sustrato. Las especies más abundantes fueron Desmapsamma anchorata, Amphimedon erina y Scopalina rueztleri. El análisis de componentes principales arrojó una separación de esta comunidad en 3 zonas, la somera (1 y 3m), donde las esponjas están sometidas a una tensión producida por el oleaje y alta iluminación, y las zonas media (6, 9 y 12m) y profunda (15 y 18m), con características más favorables, dada una menor iluminación y sedimentación.

High spatial variability of coral, sponges and gorgonian assemblages in a well preserved reef

The main goal of this research was to obtain baseline field data of the composition of sponges, corals, and gorgonian assemblages that can be used as a reference for future analyses of anthropogenic impact. We tested the hypothesis that relatively homogeneous and well preserved reef units can present notable natural variability in the composition of their communities which are unassociated with changes in land proximity or a human impact gradient. Research was carried out in July 2006 at Los Colorados reef, located in the northwestern region of Pinar del Río Province, Cuba at 12 sampling stations. The biotopes selected were crest, terrace edge and spur and grove. Ecological indicators were diversity of corals, species composition, density of corals, hydrocorals, gorgonians and sponges, and density of selected coral species. A total of 2659 colonies of scleractineans corals representing 36 species were counted. The most abundant species in the crest biotope were Millepora alcicornis, Acropora palmata and Porites astreoides; in the terrace edge and spur and grove, the most abundant species were Siderastrea siderea, Stephanocoenia intersepta, Porites astreoides, Agaricia agaricites and Montastraea cavernosa. We found differences among sites for several indicators (e.g. density of corals, sponges and gorgonians and for selected species), but they could not be associated to any gradient of land influence or human impact. Therefore, sites inside a relatively homogeneous reef unit can present notable natural differences in the composition of their communities. Rev. Biol. Trop. 58 (2): 621-634. Epub 2010 June 02.

El objetivo principal de la investigación fue obtener una línea base de la composición de las comunidades de esponjas, corales y gorgonias que pueda ser utilizada como referencia para futuros análisis de impacto antrópico. Nuestra hipótesis es que un arrecife relativamente homogéneo y bien conservado, presenta una variabilidad natural notable en la composición de sus comunidades, lo cual no está asociado con cambios en la proximidad a tierra o a un gradiente de impacto humano. La investigación se llevó a cabo en julio de 2006, en 12 estaciones de muestreo ubicadas en el arrecife de Los Colorados, en la región noroccidental de la Provincia de Pinar del Río, Cuba. Los biotopos seleccionados fueron cresta, veril y camellones. Los indicadores ecológicos estimados fueron: diversidad de corales, composición por especies, densidad de corales, hidrocorales, gorgonias y esponjas, y densidad de especies de corales seleccionadas. Se contaron 2 659 colonias de corales escleractíneos pertenecientes a 36 especies. Las especies más abundantes en el biotopo de cresta fueron: Millepora alcicornis, Acropora palmata y Porites astreoides; mientras que en el veril y los camellones fueron Siderastrea siderea, Stephanocoenia intersepta, Porites astreoides, Agaricia agaricites y Montastraea cavernosa. Entre sitios, se encontraron diferencias significativas estadísticamente para varios indicadores (densidad de corales, esponjas y gorgonias y densidad de especies seleccionadas), pero ello no pudo asociarse con ningún gradiente de uso de la tierra o de impacto humano. Por lo tanto, sitios dentro de un arrecife relativamente homogéneo pueden presentar diferencias notables en la composición de sus comunidades.