Exploring Individual- to Population-Level Impacts of Disease on Coral Reef Sponges: Using Spatial Analysis to Assess the Fate,Dynamics, and Transmission of Aplysina Red Band Syndrome (ARBS)

Background

Marine diseases are of increasing concern for coral reef ecosystems, but often their causes, dynamics and impacts are unknown. The current study investigated the epidemiology of Aplysina Red Band Syndrome (ARBS), a disease affecting the Caribbean sponge Aplysina cauliformis, at both the individual and population levels. The fates of marked healthy and ARBS-infected sponges were examined over the course of a year. Population-level impacts and transmission mechanisms of ARBS were investigated by monitoring two populations of A. cauliformis over a three year period using digital photography and diver-collected data, and analyzing these data with GIS techniques of spatial analysis. In this study, three commonly used spatial statistics (Ripley’s K, Getis-Ord General G, and Moran’s Index) were compared to each other and with direct measurements of individual interactions using join-counts, to determine the ideal method for investigating disease dynamics and transmission mechanisms in this system. During the study period, Hurricane Irene directly impacted these populations, providing an opportunity to assess potential storm effects on A. cauliformis and ARBS.

Results

Infection with ARBS caused increased loss of healthy sponge tissue over time and a higher likelihood of individual mortality. Hurricane Irene had a dramatic effect on A. cauliformis populations by greatly reducing sponge biomass on the reef, especially among diseased individuals. Spatial analysis showed that direct contact between A. cauliformis individuals was the likely transmission mechanism for ARBS within a population, evidenced by a significantly higher number of contact-joins between diseased sponges compared to random. Of the spatial statistics compared, the Moran’s Index best represented true connections between diseased sponges in the survey area. This study showed that spatial analysis can be a powerful tool for investigating disease dynamics and transmission in a coral reef ecosystem.     

Aplysina red band syndrome: a new threat to Caribbean sponges

A substantial and increasing number of reports have documented dramatic changes and continuing declines in Caribbean coral reef communities over the past 2 decades. To date, the majority of disease reports have focused on scleractinian corals, whereas sponge diseases have been less frequently documented. In this study, we describe Aplysina red band syndrome (ARBS) affecting Caribbean rope sponges of the genus Aplysina observed on shallow reefs in the Bahamas. Visible signs of disease presence included 1 or more rust-colored leading edges, with or without a trailing area of necrotic tissue, such that the lesion forms a contiguous band around part or all of the sponge branch. Microscopic examination of the leading edge of the disease margin indicated that a cyanobacterium was consistently responsible for the coloration. Although the presence of this distinctive coloration was used to characterize the diseased state, it is not yet known whether this cyanobacterium is directly responsible for disease causation. The prevalence of ARBS declined significantly from July to October 2004 before increasing above July levels in January 2005. Transmission studies in the laboratory demonstrated that contact with the leading edge of an active lesion was sufficient to spread ARBS to a previously healthy sponge, suggesting that the etiologic agent, currently undescribed, is contagious. Studies to elucidate the etiologic agent of ARBS are ongoing. Sponges are an essential component of coral reef communities and emerging sponge diseases clearly have the potential to impact benthic community structure on coral reefs.     

Does the skeleton of a sponge provide a defense against predatory reef fish?

Sponge tissue often contains two structural components in high concentrations: spicules of silica, and refractory fibers of protein (spongin). Some terrestrial plants contain analogous structures, siliceous inclusions and refractory lignins, that have been demonstrated to deter herbivory. We performed feeding experiments with predatory reef fish to assess the deterrent properties of the structural components of three common Caribbean demosponges, Agelas clathrodes, Ectyoplasia ferox, and Xestospongia muta. The concentrations of spicules and spongin in the tissues varied widely between the three species, but when assayed at their natural volumetric concentrations, neither spicules (all three species assayed) nor the intact spiculated spongin skeleton (A. clathrodes and X. muta assayed) deterred feeding by reef fish in aquarium or field assays using prepared foods of a nutritional quality similar to, or higher than, that of sponge tissue. Spicules deterred feeding in aquarium assays when incorporated into prepared foods of a nutritional quality lower than that of sponge tissue (15–19 times less protein), but spiculated spongin skeleton was still palatable, even in prepared foods devoid of measurable protein, and even though spicules embedded in spongin were oriented in their natural conformation. Based on comparisons of the nutritional qualities of the tissues of the three sponge species and of the prepared foods, sponge tissue would have to be much lower in food value (5 times less protein or lower) for spicules to provide an effective defense, and spicules in combination with the spongin skeleton would be unlikely to provide an effective defense regardless of the nutritional quality of the tissue. Unlike terrestrial plants, marine sponges may use silica and refractory fibers solely for structural purposes.     

Effects of a hurricane on survival and orientation of large erect coral reef sponges

In October 1988, Hurricane Joan struck reefs in the San Blas Island, Panama, where hurricanes had never been recorded. Effects on large erect sponges were dramatic. For several years before the hurricane, the three most common sponges had been studied, providing pre-storm data on population structure and dynamics. Nearly half the individuals and biomass of three species were lost in the storm. The species were not affected in the same way, even though they are all of erect branching growth forms. Iotrochota birotulata lost significantly more individuals chan Amphimedon rubens (57.6% vs 42.9%), which lost significantly more individuals than Aplysina fulva (31.6%). Patterns of biomass loss were very different, with both Iotrochota and Aplysina suffering losses of about 50%, but Amphimedon losing only 4.9%. Patterns of loss appear to be related to differences between species in the relative proportions of spicules (siliceous) and spongin (protein) in skeletal fibers and by differences in the speed and success rate of fragment reattachment. The incidence of toppling due to base failure varied among the six most common large erect sponge species, with significantly less toppling of the two species with skeletons composed solely of spongin. Clones of Iotrochota birotulata characterized by harrow branches suffered disproportionately greater losses than clones with more robust branches. The abundance of very small sponges, possibly developed from sexually produced larvae, was an order of magnitude higher after the storm than before.     

Shifts in microbial and chemical patterns within the marine sponge Aplysina aerophoba during a disease outbreak

The microbial community composition in affected and unaffected portions of diseased sponges and healthy control sponges of Aplysina aerophoba was assessed to ascertain the role of microbes in the disease process. Sponge secondary metabolites were also examined to assess chemical shifts in response to infection. The microbial profile and aplysinimine levels in unaffected tissue near the lesions closely reflected those of healthy sponge tissue, indicating a highly localized disease process. DGGE detected multiple sequences that were exclusively present in diseased sponges. Most notably, a Deltaproteobacteria sequence with high homology to a coral black band disease strain was detected in all sponge lesions and was absent from all healthy and unaffected regions of diseased sponges. Other potential pathogens identified by DGGE include an environmental Cytophaga strain and a novel Epsilonproteobacteria strain with no known close relatives. The disease process also caused a major shift in prokaryote community structure at a very high taxonomic level. Using 16S rRNA gene sequence analysis, only the diseased sponges were found to contain sequences belonging to the Epsilonproteobacteria and Firmicutes, and there was a much greater number of Bacteroidetes sequences within the diseased sponges. In contrast, only the healthy sponges contained sequences corresponding to the cyanobacteria and 'OP1' candidate division, and the healthy sponges were dominated by Chloroflexi and Gammaproteobacteria sequences. Overall bacterial diversity was found to be considerably higher in diseased sponges than in healthy sponges. These results provide a platform for future cultivation-based experiments to isolate the putative pathogens from A. aerophoba and perform re-infection trials to define the disease aetiology.     

The complete mitochondrial genome of the verongid sponge Aplysina cauliformis: implications for DNA barcoding in demosponges

DNA “barcoding,” the determination of taxon-specific genetic variation typically within a fragment of the mitochondrial cytochrome oxidase 1 (cox1) gene, has emerged as a useful complement to morphological studies, and is routinely used by expert taxonomists to identify cryptic species and by non-experts to better identify samples collected during field surveys. The rate of molecular evolution in the mitochondrial genomes (mtDNA) of nonbilaterian animals (sponges, cnidarians, and placozoans) is much slower than in bilaterian animals for which DNA barcoding strategies were developed. If sequence divergence among nonbilaterian mtDNA and specifically cox1 is too slow to generate diagnostic variation, alternative genes for DNA barcoding and species-level phylogenies should be considered. Previous study across the Aplysinidae (Demospongiae, Verongida) family of sponges demonstrated no nucleotide substitutions in the traditional cox1 barcoding fragment among the Caribbean species of Aplysina. As the mitochondrial genome of Aplysina fulva has previously been sequenced, we are now able to make the first comparisons between complete mtDNA of congeneric demosponges to assess whether potentially informative variation exists in genes other than cox1. In this article, we present the complete mitochondrial genome of Aplysina cauliformis, a circular molecule 19620 bp in size. The mitochondrial genome of A. cauliformis is the same length as is A. fulva and shows six confirmed nucleotide differences and an additional 11 potential SNPs. Of the six confirmed SNPs, NADH dehydrogenase subunit 5 (nad5) and nad2 each contain two, and in nad2 both yield amino acid substitutions, suggesting balancing selection may act on this gene. Thus, while the low nucleotide diversity in Caribbean aplysinid cox1 extends to the entire mitochondrial genome, some genes do display variation. If these represent interspecific differences, then they may be useful alternative markers for studies in recently diverged sponge clades.     

ITS-2 and 18S rRNA Gene Phylogeny of Aplysinidae (Verongida, Demospongiae)

18S ribosomal DNA and internal transcribed spacer 2 (ITS-2) full-length sequences, each of which was sequenced three times, were used to construct phylogenetic trees with alignments based on secondary structures, in order to elucidate genealogical relationships within the Aplysinidae (Verongida). The first poriferan ITS-2 secondary structures are reported. Altogether 11 Aplysina sponges and 3 additional sponges (Verongula gigantea, Aiolochroia crassa, Smenospongia aurea) from tropical and subtropical oceans were analyzed. Based on these molecular studies, S. aurea, which is currently affiliated with the Dictyoceratida, should be reclassified to the Verongida. Aplysina appears as monophyletic. A soft form of Aplysina lacunosa was separated from other Aplysina and stands at a basal position in both 18S and ITS-2 trees. Based on ITS-2 sequence information, the Aplysina sponges could be distinguished into a single Caribbean–Eastern Pacific cluster and a Mediterranean cluster. The species concept for Aplysina sponges as well as a phylogenetic history with a possibly Tethyan origin is discussed.Reviewing Editor: Dr. Martin Kreitman     

The iron mineralization of Spongia officinalis L. (Porifera,Dictyoceratida) and its relationships with the collagen skeleton

Lepidocrocite (γ FeOOH) granules are frequently found in the spongin fibers of some horny sponges. The granules also contain calcite and a small amount of goethite (α FeOOH). In the genus Spongia, the amount of iron is highly variable according to the habitat and to the age of the fiber. Some indications about the skeletal growth and structure can be inferred from the distribution of the biominerals and the arrangement of the crystallites: i.e. (1) Mineralization is a slow process in comparison with fiber formation; (2) Secondary connective fibers may have a discontinuous growth; (3) The collagen microfibrils display a definite arrangement in the successive spongin layers; (4) Although spongin is a collagen highly resistant in vitro to enzymatic digestion, the sponge is able to perform remodeling of its skeleton through autodigestion of the fibers.     

Asterocherids (Copepoda; Siphonostomatoida) associated with invertebrates from California Reefs: Abrolhos (Brazil)

Four new asterocherid siphonostomatoid copepods are described, and several new geographical and host records are provided from California Reefs off the coast of Brazil. Acontiophorus monanchorae sp. nov. and Scottocheres youngi sp. nov. were both found associated with the sponge Monanchora sp. Asterocheres aplysinus sp. nov. and Asterocheres spongus sp. nov. were each associated with three genera of sponges. Asterocheres crenulatus previously recorded from the region, was associated with the sponge Xestospongia sp. Asterocheres paraboecki was recorded for the first time in the region. Asterocheres spinopaulus, previously reported from sponges, was found associated with bryozoans and cnidarians. Cletopontius titanus and Orecturus bahiensis were again recorded from the region and from sponges. Scottocheres laubieri, previously known from the Mediterranean Sea, was recorded for the first time in Brazilian waters, associated with the sponge Aplysina sp.     

Leptothoe,a new genus of marine cyanobacteria (Synechococcales) and three new species associated with sponges from the Aegean Sea

Cyanobacterial diversity associated with sponges remains underestimated, though it is of great scientific interest in order to understand the ecology and evolutionary history of the symbiotic relationships between the two groups. Of the filamentous cyanobacteria, the genus Leptolyngbya is the most frequently found in association with sponges as well as the largest and obviously polyphyletic group. In this study, five Leptolyngbya‐like sponge‐associated isolates were investigated using a combination of molecular, chemical, and morphological approach and revealed a novel marine genus herein designated Leptothoe gen. nov. In addition, three new species of Leptothoe, Le. sithoniana, Le. kymatousa, and Le. spongobia, are described based on a suite of distinct characters compared to other marine Leptolyngbyaceae species/strains. The three new species, hosted by four sponge species, showed different degrees of host specificity. Leptothoe sithoniana and Le. kymatousa hosted by the sponges Petrosia ficiformis and Chondrilla nucula, respectively, seem to be more specialized than Le. spongobia, which was hosted by the sponges Dysidea avara and Acanthella acuta. All three species contained nitrogen‐fixing genes and may contribute to the nitrogen budget of sponges. Leptothoe spongobia TAU‐MAC 1115 isolated from Acanthella acuta was shown to produce microcystin‐RR indicating that microcystin production among marine cyanobacteria could be more widespread than previously determined.     

Phylogenetic analyses of marine sponges within the order Verongida: a comparison of morphological and molecular data

Abstract. Because the taxonomy of marine sponges is based primarily on morphological characters that can display a high degree of phenotypic plasticity, current classifications may not always reflect evolutionary relationships. To assess phylogenetic relationships among sponges in the order Verongida, we examined 11 verongid species, representing six genera and four families. We compared the utility of morphological and molecular data in verongid sponge systematics by comparing a phylogeny constructed from a morphological character matrix with a phylogeny based on nuclear ribosomal DNA sequences. The morphological phylogeny was not well resolved below the ordinal level, likely hindered by the paucity of characters available for analysis, and the potential plasticity of these characters. The molecular phylogeny was well resolved and robust from the ordinal to the species level. We also examined the morphology of spongin fibers to assess their reliability in verongid sponge taxonomy. Fiber diameter and pith content were highly variable within and among species. Despite this variability, spongin fiber comparisons were useful at lower taxonomic levels (i.e., among congeneric species); however, these characters are potentially homoplasic at higher taxonomic levels (i.e., between families). Our molecular data provide good support for the current classification of verongid sponges, but suggest a re-examination and potential reclassification of the genera Aiolochroia and Pseudoceratina . The placements of these genera highlight two current issues in morphology-based sponge taxonomy: intermediate character states and undetermined character polarity.     

Chemical defense of Mediterranean sponges Aplysina cavernicola and Aplysina aerophoba

The Mediterranean sponges Aplysina aerophoba and A. cavernicola accumulate brominated isoxazoline alkaloids including aplysinamisin-1 (1), aerophobin-2 (2), isofistularin-3 (3) or aerothionin (4) at concentrations up to 10% of their respective dry weights. In laboratory feeding experiments employing the polyphagous Mediterranean fish Blennius sphinx crude extracts of both Aplysina sponges were incorporated into artificial fish food at their physiological concentrations (based on volume) and offered to B. sphinx in choice feeding experiments against untreated control food. In addition to the Aplysina sponges, extracts from nine other frequently occurring Mediterranean sponges were likewise included into the experiments. Both Aplysina species elicited strong feeding deterrence compared to the other sponges tested. Bioassay-guided fractionation of A. cavernicola yielded the isoxazoline alkaloids aerothionin (4) and aplysinamisin-1 (1) as well as the 3,4-dihydroxyquinoline-2-carboxylic acid (8) as major deterrent constituents when tested at their physiological concentrations as present in sponges. Aeroplysinin-1 (5) and dienone (6), however, which are formed in A. aerophoba and A. cavernicola from isoxazoline precursors through bioconversion reactions upon tissue injury showed no or only little deterrent activity. Fractionation of a crude extract of A. aerophoba yielded aerophobin-2 (2) and isofistularin-3 (3) as major deterrent constituents against B. sphinx. We propose that the isoxazoline alkaloids 1-4 of Mediterranean Aplysina sponges as well as the 3,4-dihydroxyquinoline-2-carboxylic acid (8) (in the case of A. cavernicola) act as defensive metabolites against B. sphinx and possibly also against other predators while the antibiotically active bioconversion products aeroplysinin-1 (5) and dienone (6) may protect sponges from invasion of bacterial pathogens.     

Thermostablility of phycobiliproteins and antioxidant activity from four thermotolerant cyanobacteria

Four cyanobacterial strains including Cyanosarcina sp. SK40, Phormidium sp. PD40‐1, Scytonema sp. TP40 and Leptolyngbya sp. KC45 were selected and investigated for the phycobiliprotein (PBP) content and thermostable antioxidant activity of their cell‐free extracts. The highest content of 181.63 mg/g dry weight phycobiliprotein was found in Leptolyngbya sp. KC45 with phycoerythrin (PE) as the main phycobiliprotein. Among the PBPs of four thermotolerant cyanobacteria, PE from Leptolyngbya sp. KC45 exhibited the highest thermal stability as 80% of the original level remained after being heated at 60°C for 30 min. Antioxidant activities were detected in the cell‐free extracts of all cyanobacteria and that of Leptolyngbya sp. KC45 was also found in the highest value of 7.44 ± 0.14 and 3.89 ± 0.08 mg gallic acid equivalent (GAE) g^?1 dry weights determined by 2,2‐diphenyl‐1‐picrylhydrazyl radical (DPPH) and reducing power assay, respectively. This also corresponded to the phenolic compound content. Based on DPPH and reducing power assay, antioxidant activities of all cyanobacterial extracts showed the high thermostability as approximately 80% remained after being heated at 80°C for 30 min. However, it clearly indicated that the thermostability of antioxidant activity from the hot spring cyanobacterial cell‐free extract was not contributed only by the PE, but also came from phenolic compounds and other oxidative substances.     

Pyrosequencing Reveals the Microbial Communities in the Red Sea Sponge Carteriospongia foliascens and Their Impressive Shifts in Abnormal Tissues

Abnormality and disease in sponges have been widely reported, yet how sponge-associated microbes respond correspondingly remains inconclusive. Here, individuals of the sponge Carteriospongia foliascens under abnormal status were collected from the Rabigh Bay along the Red Sea coast. Microbial communities in both healthy and abnormal sponge tissues and adjacent seawater were compared to check the influences of these abnormalities on sponge-associated microbes. In healthy tissues, we revealed low microbial diversity with less than 100 operational taxonomic units (OTUs) per sample. Cyanobacteria, affiliated mainly with the sponge-specific species “Candidatus Synechococcus spongiarum,” were the dominant bacteria, followed by Bacteroidetes and Proteobacteria. Intraspecies dynamics of microbial communities in healthy tissues were observed among sponge individuals, and potential anoxygenic phototrophic bacteria were found. In comparison with healthy tissues and the adjacent seawater, abnormal tissues showed dramatic increase in microbial diversity and decrease in the abundance of sponge-specific microbial clusters. The dominated cyanobacterial species Candidatus Synechococcus spongiarum decreased and shifted to unspecific cyanobacterial clades. OTUs that showed high similarity to sequences derived from diseased corals, such as Leptolyngbya sp., were found to be abundant in abnormal tissues. Heterotrophic Planctomycetes were also specifically enriched in abnormal tissues. Overall, we revealed the microbial communities of the cyanobacteria-rich sponge, C. foliascens, and their impressive shifts under abnormality.     

Platelet Aggregation Inhibitors from Philippine Marine Invertebrate Samples Screened in a New Microplate Assay

A new microplate assay for Ca²⁺-induced platelet aggregation as detected by Giemsa dye was used to screen marine invertebrate samples from the Philippines for inhibitors of human platelet aggregation. Out of 261 crude methanol extracts of marine sponges and tunicates, 25 inhibited aggregation at 2 mg/ml. Inhibition of agonist-induced aggregation in an aggregometer was used to confirm results of the microplate assay and to determine the specific mode of inhibition of 2 samples. The marine sponge Xestospongia sp. yielded a xestospongin/araguspongine-type molecule that inhibited collagen-induced aggregation by 87% at 2 µg/ml, and epinephrine-induced aggregation by 78% at 20 µg/ml, while the marine sponge Aplysina sp. yielded 5,6-dibromotryptamine, which inhibited epinephrine-induced aggregation by 51% at 20 µg/ml. In this study we have found that the microplate assay is a simple, inexpensive, yet useful preliminary tool to qualitatively screen a large number of marine samples for antiplatelet aggregation activity.     

Soft sponges with tricky tree: On the phylogeny of dictyoceratid sponges

Keratose (horny) sponges constitute a very difficult group of Porifera in terms of taxonomy due to their paucity of diagnostic morphological features. (Most) keratose sponges possess no mineral skeletal elements, but an arrangement of organic (spongin) fibers, with little taxonomic or phylogenetic information. Molecular phylogenetics have targeted this evolutionary and biochemically important lineage numerous times, but the conservative nature of popular markers combined with ambiguous identification of the sponge material has so far prevented any robust phylogeny. In the following study, we provide a phylogenetic hypothesis of the keratose order Dictyoceratida based on nuclear markers of higher resolution potential (ITS and 28S C-region), and particularly aim for the inclusion of type specimens as reference material. Our results are compared with previously published data of CO1, 18S, and 28S (D3-D5) data, and indicate the paraphyly of the largest dictyoceratid family, the Thorectidae, due to a sister group relationship of its subfamily Phyllospongiinae with Family Spongiidae. Irciniidae can be recovered as monophyletic. Results on genus level and implications on phylogenetic signals of the most frequently described morphological characters are discussed.     

Bioactive Brominated Metabolites from the Natural Habitat and Tank‐Maintained Cuttings of the Jamaican Sponge Aplysina fistularis

Cut specimens of the common reef sponge of the Verongid family, Aplysina fistularis, were retained in flow‐through seawater tanks over a six‐week period to assess the metabolite profile of the sponge when subjected to stress, compare the profile with the source material, and assess the preliminary feasibility of the protocol for sponge culture. The living specimens were harvested, extracted with MeOH/CH₂Cl₂ 1 : 1, and subjected to column chromatography to identify metabolites. The brominated isoxazoline compounds, aerothionin ( 1 ) and 11‐oxoaerothionin ( 2 ), along with aeroplysinin 2 ( 3 ) and 2‐(3,5‐dibromo‐4‐hydroxyphenol)acetamide ( 4 ), were detected in the cuttings from the tank‐maintained sponge. An examination of the metabolite profile of the sponge from the natural habitat showed that the compounds 1 and 2 were present. The identities of all the compounds were ascertained by analysis of the mass‐spectral data and NMR spectra (¹H, ¹³C, HMBC, and HSQC) of the compounds, which were compared with reported data. The survival rate was 44% with limited necrosis or exposed skeletal tissue being observed in eight of the 18 cuttings, suggesting that protocol modifications would be required for culturing the sponge.     

Identification of Planctomycetes with Order-, Genus-, and Strain-Specific 16S rRNA-Targeted Probes

A specific 16S rRNA-targeted oligonucleotide probe (PIR1223) for the genus Pirellula and a species-specific probe (RB454) for Pirellula sp. strain SH1 have been designed and optimized. Together with the already existing order-specific probe PLA886, the two newly designed probes were used to detect and identify planctomycetes, pirellulae, and close relatives of Pirellula sp. strain SH1 in different habitats. With the help of these probes for detection and identification, bacteria of the genus Pirellula were detected and cultivated from tissue of the Mediterranean sponge Aplysina aerophoba and from the water column of the Kiel Fjord. An unexpected result was the close phylogenetic relationship of the isolate from the sponge and the brackish water habitat Kiel Fjord as revealed by DNA/DNA hybridization.     

Isolation and phylogenetic analysis of bacteria with antimicrobial activities from the Mediterranean sponges Aplysina aerophoba and Aplysina cavernicola

The aim of this study was to isolate bacteria with antimicrobial activities from the marine sponges Aplysina aerophoba and Aplysina cavernicola. The obtained 27 isolates could be subdivided into eight phylogenetically different clusters based on comparative sequence analysis of their 16S rDNA genes. The sponge isolates were affiliated with the low (Bacillus) and high G+C Gram-positive bacteria (Arthobacter, Micrococcus), as well as the alpha-Proteobacteria (unknown isolate) and gamma-Proteobacteria (Vibrio, Pseudoalteromonas). One novel Bacillus species was identified and two species were closely related to previously uncharacterized strains. Isolates with antimicrobial activity were numerically most abundant in the genera Pseudoalteromonas and the alpha-Proteobacteria. The sponge isolates show antimicrobial activities against Gram-positive and Gram-negative reference strains but not against the fungus Candida albicans. A general pattern was observed in that Gram-positive bacteria inhibited Gram-positive strains while Gram-negative bacteria inhibited Gram-negative isolates. Antimicrobial activities were also found against clinical isolates, i.e. multi-resistant Staphylococcus aureus and Staphylococcus epidermidis strains isolated from hospital patients. The high recovery of strains with antimicrobial activity suggests that marine sponges represent an ecological niche which harbors a hitherto largely uncharacterized microbial diversity and, concomitantly, a yet untapped metabolic potential.     

Cyanobacterial diversity in extreme environments in Baja California, Mexico: a polyphasic study

Cyanobacterial diversity from two geographical areas of Baja California Sur, Mexico, were studied: Bahia Concepcion, and Ensenada de Aripez. The sites included hypersaline ecosystems, sea bottom, hydrothermal springs, and a shrimp farm. In this report we describe four new morphotypes, two are marine epilithic from Bahia Concepcion, Dermocarpa sp. and Hyella sp. The third, Geitlerinema sp., occurs in thermal springs and in shrimp ponds, and the fourth, Tychonema sp., is from a shrimp pond. The partial sequences of the 16S rRNA genes and the phylogenetic relationship of four cyanobacterial strains (Synechococcus cf. elongatus, Leptolyngbya cf. thermalis, Leptolyngbya sp., and Geitlerinema sp.) are also presented. Polyphasic studies that include the combination of light microscopy, cultures and the comparative analysis of 16S rRNA gene sequences provide the most powerful approach currently available to establish the diversity of these oxygenic photosynthetic microorganisms in culture and in nature. Electronic Publication