Diseased Sponges Research Articles

Mass mortality of the keratose sponge Sarcotragus foetidus in the Aegean Sea (Eastern Mediterranean) correlates with proliferation of Vibrio bacteria in the tissues.

In the last two decades, episodes of mass mortality in benthic communities have often been associated with climatic anomalies, but the ultimate mechanisms through which they lead to death have rarely been identified. This study reports a mass mortality of wild sponges in the Aegean Sea (Turkey, Eastern Mediterranean), which affected the keratose demosponge Sarcotragus foetidus in September 2021. We examined the occurrence of thermo-dependent bacteria of the genus Vibrio in the sponges, identified through 16S rRNA of colonies isolated from sponge tissue in specific culturing media. Six Vibrio sequences were identified from the sponges, three of them being putatively pathogenic (V. fortis, V. owensii, V. gigantis). Importantly, those Vibrios were isolated from only tissues of diseased sponges. In contrast, healthy individuals sampled in both summer and winter led to no Vibrio growth in laboratory cultures. A 50 years record of sea surface temperature (SST) data for the study area reveals a progressive increase in temperature from 1970 to 2021, with values above 24°C from May to September 2021, reaching an absolute historical maximum of 28.9°C in August 2021. We hypothesize that such elevated SST values maintained for several months in 2021 promoted proliferation of pathogenic Vibrio species (thermo-dependent bacteria) in S. foetidus, triggering or aggravating the course of sponge disease. Thus, vibrioisis emerges as one of the putative mechanisms through which global water warming in the Mediterranean Sea translates into sponge mortality. The historical time course of temperature data for the studied area in the Aegean Sea predicts that recurrent waves of elevated SST are likely to occur in the coming summers. If so, recurrent disease may eventually eliminate this abundant sponge from the sublittoral in the midterm, altering the original bathymetric distribution of the species and compromising its ecological role.

COMPOSITION AND DISTRIBUTION OF MACROINVERTEBRATES ASSOCIATED WITH <i>LUBOMIRSKIA BAIKALENSIS</i> SPONGES (SPONGILLIDA, LUBOMIRSKIIDAE) DURING AN ECOLOGICAL CRISIS IN LAKE BAIKAL

This study was carried out in the littoral zone of Lake Baikal during a large-scale ecological crisis that affected all parts of the ecosystem, including populations of endemic sponges of the family Lubomirskiidae. Sponges play important roles in biotic communities of the rocky littoral of the lake, ensuring water purity and serving as a specialized habitat for a variety of organisms. In the last decade, mass disease and mortality of sponges were reported in different areas of Lake Baikal, which can lead to irreversible changes in the structure of littoral communities. We studied the taxonomic richness, distribution, and abundance of the main groups of macroinvertebrates, such as amphipods and gastropods, living on diseased branched Lubomirskia baikalensis sponges in three basins of Lake Baikal (2015, 2020, 2021, 2022) and made historic comparisons. The taxonomic richness of amphipods, including 35 species or subspecies, as well as further 5 taxa identified only to genus, increased and had 20 taxa more than 30 years ago. A total of 22 gastropod species or subspecies were found on the sponge, 20 of them for the first time. The species composition, dominant species complex and quantitative characteristics of sponge-associated amphipods and gastropods varied in different lake areas as evidenced by underwater observations. The abundance of all macroinvertebrate groups, as well as dominant amphipod and gastropod groups per unit weight of sponges decreased from south to north. The abundance of the invertebrates associated with diseased sponges was similar to the records of the pre-crisis period. Baikal sponges host invertebrates from several ecological groups with different feeding strategies (grazers, scrapers and filter feeders) providing a temporary refuge for most of them. In order to restore the abundance of sponges and preserve the populations of key invertebrate species, we propose to set up sponge “farms” in the least polluted areas of Lake Baikal.

Janthinobacterium sp. Strain SLB01 as Pathogenic Bacteria for Sponge Lubomirskia baikalensis

Sponges (phylum Porifera) are ancient, marine and inland water, filter feeding metazoans. In recent years, diseased sponges have been increasingly occurring in marine and freshwater environments. Endemic freshwater sponges of the Lubomirskiidae family are widely distributed in the coastal zone of Lake Baikal. The strain Janthinobacterium sp. SLB01 was isolated previously from the diseased sponge Lubomirskia baikalensis (Pallas, 1776), although its pathogenicity is still unknown. The aim of this study was to confirm whether the Janthinobacterium sp. strain SLB01 is the pathogen found in Baikal sponge. To address this aim, we infected the cell culture of primmorphs of the sponge L. baikalensis with strain SLB01 and subsequently reisolated and sequenced the strain Janthinobacterium sp. PLB02. The results showed that the isolated strain has more than 99% homology with strain SLB01. The genomes of both strains contain genes vioABCDE of violacein biosynthesis and floc formation, for strong biofilm, in addition to the type VI secretion system (T6SS) as the main virulence factor. Based on a comparison of complete genomes, we showed the similarity of the studied bacterial strains of Janthinobacterium spp. with the described strain of Janthinobacterium lividum MTR. This study will help expand our understanding of microbial interactions and determine one of the causes in the development of diseases and death in Baikal sponges.

Transmission studies and the composition of prokaryotic communities associated with healthy and diseased Aplysina cauliformis sponges suggest that Aplysina Red Band Syndrome is a prokaryotic polymicrobial disease.

Aplysina cauliformis, the Caribbean purple rope sponge, is commonly affected by Aplysina Red Band Syndrome (ARBS). This transmissible disease manifests as circular lesions with red margins and results in bare spongin fibers. Leptolyngbya spp. appear to be responsible for the characteristic red coloration but transmission studies with a sponge-derived isolate failed to establish disease, leaving the etiology of ARBS unknown. To investigate the cause of ARBS, contact transmission experiments were performed between healthy and diseased sponges separated by filters with varying pore sizes. Transmission occurred when sponges were separated by filters with pore sizes ≥ 2.5 μm, suggesting a prokaryotic pathogen(s) but not completely eliminating eukaryotic pathogen(s). Using 16S rRNA gene sequencing methods, 38 prokaryotic taxa were significantly enriched in diseased sponges, including Leptolyngbya, whereas seven taxa were only found in some, but not all, of the ARBS-affected sponges. These results do not implicate a single taxon, but rather a suite of taxa that changed in relative abundance with disease, suggesting a polymicrobial etiology as well as dysbiosis. As a better understanding of dysbiosis is gained, changes in the composition of associated prokaryotic communities may have increasing importance for evaluating and maintaining the health of individuals and imperiled coral reef ecosystems.

Genome Analysis of the Janthinobacterium sp. Strain SLB01 from the Diseased Sponge of the Lubomirskia baicalensis.

The strain Janthinobacterium sp. SLB01 was isolated from the diseased freshwater sponge Lubomirskia baicalensis (Pallas, 1776) and the draft genome was published previously. The aim of this work is to analyze the genome of the Janthinobacterium sp. SLB01 to search for pathogenicity factors for Baikal sponges. We performed genomic analysis to determine virulence factors, comparing the genome of the strain SLB01 with genomes of other related J. lividum strains from the environment. The strain Janthinobacterium sp. SLB01 contained genes encoding violacein, alpha-amylases, phospholipases, chitinases, collagenases, hemolysin, and a type VI secretion system. In addition, the presence of conservative clusters of genes for the biosynthesis of secondary metabolites of tropodithietic acid and marinocine was found. We present genes for antibiotic resistance, including five genes encoding various lactamases and eight genes for penicillin-binding proteins, which are conserved in all analyzed strains. Major differences were found between the Janthinobacterium sp. SLB01 and J. lividum strains in the spectra of genes for glycosyltransferases and glycoside hydrolases, serine hydrolases, and trypsin-like peptidase, as well as some TonB-dependent siderophore receptors. Thus, the study of the analysis of the genome of the strain SLB01 allows us to conclude that the strain may be one of the pathogens of freshwater sponges.

Microbiome Changes of Endemic Lake Baikal Sponges during Bleaching Syndrome Development

The sponge (Porifera) microbiome is an indicator of both natural and anthropogenic stressors. Studying Baikal sponge microbial communities could help reveal if there is a connection between bacterial symbionts and a mass sponge bleaching event that was recently detected; 16S rRNA sequencing was performed among healthy and diseased freshwater sponges of Lubomirskia baikalensis and Baikalospongia intermedia, which were collected from Lake Baikal, Russia. A phylum-based taxonomic classification showed that Chlorophyta, Acidobacteria, Bacteroidetes, Actinobacteria and Cyanobacteria were most abundant across samples. When comparing healthy and diseased L. baikalensis samples, large variations in microbial composition were found at the phylum level. Comparative analyses, which were performed for the first time for B. intermedia, showed a decrease in Chlorophyta (unicellular green algae) and an increase in Bacteroidetes and Cyanobacteria in diseased specimens. At the genus level, the Opitutus (Verrucomicrobia), Planctomyces, and Nitrospira content increased in all diseased sponges, which reflected a general tendency toward an increase in Cyanobacteria in diseased sponges. Comparative analysis of the diseased and healthy sponge metagenomes showed that diseased sponges underwent various nonspecific changes in bacterial composition. The bacterial community composition is probably influenced by sponge type and degree of disease affection.

Baikal endemic sponge disease and anthropogenic factor

This article presents the results of observations of the state of the Baikal endemic sponge fauna (Fam. Lubomirskiidae) in the five-year dynamics against the background of their mass mortality. We indicate the change in the projective bottom cover with healthy and diseased sponges at two sites that are radically different from each other in the level of anthropogenic pressure. We also analyse the types of diseases and sponge species exposed to these at the study sites. We observe a significant decrease in the area of the projective bottom cover with healthy sponges both at the site experiencing heavy anthropogenic pressure and at the site where it is almost absent. However, our data have indicated great influence of the anthropogenic factor on the course of the disease and death of sponges. Degradation of sponges at the anthropogenically unfavourable site began much earlier than that at the site with the minimum anthropogenic impact. The most common species are the most susceptible to all types of diseases. At the same time, there is a catastrophic decrease in the abundance of the only branching species in Lake Baikal, Lubomirskia baikalensis.

Cooperative Interaction of Janthinobacterium sp. SLB01 and Flavobacterium sp. SLB02 in the Diseased Sponge Lubomirskia baicalensis.

Endemic freshwater sponges (demosponges, Lubomirskiidae) dominate in Lake Baikal, Central Siberia, Russia. These sponges are multicellular filter-feeding animals that represent a complex consortium of many species of eukaryotes and prokaryotes. In recent years, mass disease and death of Lubomirskia baicalensis has been a significant problem in Lake Baikal. The etiology and ecology of these events remain unknown. Bacteria from the families Flavobacteriaceae and Oxalobacteraceae dominate the microbiomes of diseased sponges. Both species are opportunistic pathogens common in freshwater ecosystems. The aim of our study was to analyze the genomes of strains Janthinobacterium sp. SLB01 and Flavobacterium sp. SLB02, isolated from diseased sponges to identify the reasons for their joint dominance. Janthinobacterium sp. SLB01 attacks other cells using a type VI secretion system and suppresses gram-positive bacteria with violacein, and regulates its own activity via quorum sensing. It produces floc and strong biofilm by exopolysaccharide biosynthesis and PEP-CTERM/XrtA protein expression. Flavobacterium sp. SLB02 utilizes the fragments of cell walls produced by polysaccharides. These two strains have a marked difference in carbohydrate acquisition. We described a possible means of joint occupation of the ecological niche in the freshwater sponge microbial community. This study expands the understanding of the symbiotic relationship of microorganisms with freshwater Baikal sponges.

Draft Genome Sequence of Flavobacterium sp. Strain SLB02, Isolated from the Diseased Sponge Lubomirskia baicalensis.

There are significant changes in the consortium of microorganisms of freshwater Baikal sponges during their mass death, which began in 2011. The alleged cause of disease is a significant increase in the number of opportunistic microorganisms. Here, we report the draft genome sequence of Flavobacterium sp. strain SLB02.

Microbiome analysis of healthy and diseased sponges Lubomirskia baicalensis by using cell cultures of primmorphs.

Endemic sponges (Demosponges, Lubomirskiidae) dominate the fauna of the littoral zone of Lake Baikal. These freshwater sponges live in symbiosis with diverse eukaryotes and prokaryotes, including chlorophyll-containing microalgae. Within the last 5 years, the incidence of sponge disease and mortality events in Lake Baikal has increased. The etiology and ecology of these events remain unknown, in part because of the lack of models to study sponge-microbe interactions. In this work, we tested the use of primmorph cell cultures of Lubomirskia baicalensis as a tool for investigating the microbiomes of sponges. We infected primmorphs, cultured in vitro, with samples from diseased sponges and observed, by microscopy, disease symptoms, including loss of green symbionts, associated with mass die-off events. Subsequent sequencing of 16S rRNA gene fragments revealed that the microbiome community of healthy sponge and primmorphs formed a group separate from the community of diseased sponges and infected primmorphs. This confirms the suitability of the primmorph cell culture as a model sponge system. We also discovered mass mortality of green symbionts (Chlorophyta) was associated with a shift in the microbial communities of sponges/primmorphs. Microbes in diseased sponges, and infected primmorphs, belonged mainly to the phyla Bacteroidetes and Proteobacteria and these families Flavobacteriaceae, Burkholderiaceae, and Moraxellaceae. Primmorphs cell culture may provide a model to study interactions between these bacteria and their host and elucidate the cause of mass mortality events.

The signs of adaptive mutations identified in the chloroplast genome of the algae endosymbiont of Baikal sponge.

Monitoring and investigating the ecosystem of the great lakes provide a thorough background when forecasting the ecosystem dynamics at a greater scale. Nowadays, changes in the Baikal lake biota require a deeper investigation of their molecular mechanisms. Understanding these mechanisms is especially important, as the endemic Baikal sponge disease may cause a degradation of the littoral ecosystem of the lake. The chloroplast genome fragment for the algae endosymbiont of the Baikal sponge was assembled from metagenomic sequencing data. The distributions of the polymorphic sites were obtained separately for the genome fragments from healthy, diseased and dead sponge tissues. The distribution of polymorphic sites allows for the detection of the signs of extensive mutations in the chloroplasts isolated from the diseased sponge tissues. Additionally, the comparative analysis of chloroplast genome sequences suggests that the symbiotic algae from Baikal sponge is close to the Choricystis genus of unicellular algae. Mutations observed in the chloroplast genome could be interpreted as signs of rapid adaptation processes in the symbiotic algae. The development of sponge disease is still expanding in Baikal, but an optimistic prognoses regarding a development of the disease is nevertheless considered.

The signs of adaptive mutations identified in the chloroplast genome of the algae endosymbiont of Baikal sponge.

Background: Monitoring and investigating the ecosystem of the great lakes provide a thorough background when forecasting the ecosystem dynamics at a greater scale. Nowadays, changes in the Baikal lake biota require a deeper investigation of their molecular mechanisms. Understanding these mechanisms is especially important, as the endemic Baikal sponge disease may cause a degradation of the littoral ecosystem of the lake. Methods: The chloroplast genome fragment for the algae endosymbiont of the Baikal sponge was assembled from metagenomic sequencing data. The distributions of the polymorphic sites were obtained separately for the genome fragments from healthy, diseased and dead sponge tissues. Results: The distribution of polymorphic sites allows for the detection of the signs of extensive mutations in the chloroplasts isolated from the diseased sponge tissues. Additionally, the comparative analysis of chloroplast genome sequences suggests that the symbiotic algae from Baikal sponge is close to the Choricystis genus of unicellular algae. Conclusions: Mutations observed in the chloroplast genome could be interpreted as signs of rapid adaptation processes in the symbiotic algae. The development of sponge disease is still expanding in Baikal, but an optimistic prognoses regarding a development of the disease is nevertheless considered.

First data on cyanobacterial biodiversity in benthic biofilms during mass mortality of endemic sponges in Lake Baikal

This study investigates in detail species diversity of cyanobacteria which proliferated on the surface of diseased sponges and bedrock in Lake Baikal (Siberia, Russia) during the ecological crisis that has been progressing in the littoral zone since 2011 and caused numerous negative changes in biota. In 2014–2015, we examined samples from diseased and healthy sponges as well as rock substrates using Sanger sequencing and high-throughput 16S rRNA gene analysis. Based on morphology, we identified 15 cyanobacterial species in biofoulings. Tychonema sp., Symplocastrum sp., Pseudanabaena galeata and Tolypothrix distorta dominated on diseased sponges. In communities of healthy sponges, we detected mainly picocyanobacteria Synechococcus. Fascicles of Symplocastrum sp. and T. distorta dominated on bedrock substrates. Phylogenetically, most 16S rDNAs clustered among Tychonema, Pseudanabaena, Geitlerinema, Kamptonema, Leptolyngbya, and Tolypothrix spp. Cyanobacterial proliferation in the littoral zone of Lake Baikal should be considered as one of the most important negative ecological processes in recent times which requires an urgent study in order to understand the causes of its emergence and development. Mat-forming cyanobacteria Tychonema characterized by the highest tropism to sponges plays a special role in the disease and death of sponges.

Draft Genome Sequence of Janthinobacterium sp. Strain SLB01, Isolated from the Diseased Sponge Lubomirskia baicalensis.

The draft genome sequence of Janthinobacterium sp. strain SLB01, a violacein-producing psychrotolerant bacterium isolated from the diseased sponge Lubomirskia baicalensis, was determined. We identified five genes encoding VioA, VioB, VioC, VioD, and VioE proteins related to violacein biosynthesis that were like those identified in published Janthinobacterium lividum strains MTR and RIT308.

Variation of sponge-inhabiting infauna with the state of health of the sponge Lubomirskia baikalensis (Pallas, 1776) in Lake Baikal

Our investigation was conducted during a period of ecological crisis in the coastal zone of Lake Baikal. Mass disease and mortality of the endemic sponges inhabiting the nearshore zone of the lake is one of the characteristics of this crisis. We identified and quantified infaunal organisms associated with Lubomirskia baikalensis (Pallas, 1776), which is experiencing mass morbidity and mortality. L. baikalensis specimens were subdivided into three groups depending on the degree of body damage they presented: “healthy”, diseased, or dead. We found that infauna was almost absent from “healthy” sponges. Infaunal abundance in diseased sponges was 1820 times greater than that in “healthy” sponges, and varied in relation to the type of damage suffered by the sponges. Unaffected fragments of diseased sponges were inhabited by communities that exhibited an average abundance of 13 ind/dm2, with Oligochaeta, Nematoda, Harpacticoida, and Chironomidae predominating. In the bleached fragments of morbid sponges, the average abundance of infauna was 1303 ind/dm2, with Harpacticoida, Tardigrada, Cyclopoida, and Oligochaeta dominating. The highest concentration of infaunal animals (18,293 ind/dm2) was observed in spots that were covered by filamentous cyanobacteria. Diverse and densely populated infaunal communities (4767 ind/dm2) occurred in the dead sponges, with Nematoda, Tardigrada, Turbellaria, and Oligochaeta particularly abundant.

Diversity and shifts of the bacterial community associated with Baikal sponge mass mortalities.

The disease of freshwater sponges was first discovered in 2011, when pink samples were found in the Central Basin of Lake Baikal. Subsequently, the visible signs of the disease have changed, and now sponges appear with various symptoms of damage to the body, such as discoloration, tissue necrosis, the formation of brown patches and dirty-purple biofilms on some branches. These signs of the disease are accompanied by the mass death of sponges. We identified differences in microbiomes by sequencing 16S rRNA genes and found changes in the consortium of microorganisms of freshwater Baikal sponges. We found that the observed imbalance in the studied microbial communities of diseased sponges is caused by several different conditionally pathogenic microorganisms that increase their negative effect by acting together and in concert, which leads to the death of photosynthetic microalgae and sponges. Sponges are an important component of coastal communities, and the massive loss of sponges can obviously affect the structure of benthic communities and the purity of water.

Diet of the Symbiotic Amphipod, Brandtia parasitica parasitica (Crustacea, Amphipoda), Living on Diseased Baikal Sponges of the Family Lubomirskiidae in Southern Lake Baikal

The diet of a symbiont, Brandtia parasitica parasitica (Dybowsky 1874), of Baikal sponges collected during an ecological crisis in the littoral zone of Lake Baikal is analyzed. In June 2015, the main components of the diet of the amphipod that lived on diseased sponges were fine detritus, sedimented chrysophycean algae of the genus Dinobryon, and planktonic diatoms of the genus Synedra. These algae dominate in phytoplankton in the southern part of Baikal during the study period. Abundant blue-green algae were present on the surface of diseased sponges, but B. p. parasitica did not feed on these algae. As toxins secreted by blue-green algae are known to render a negative effect on hydrobionts, mass overgrowth of sponges with blue-green algae should reduce the population of the symbiotic amphipod.

The signs of adaptive mutations identified in the chloroplast genome of the algae endosymbiont of Baikal sponge.

Background: The study of ecosystems of the great lakes is important as observations can be extended to ecosystems of larger scale. The ecological crisis of Lake Baikal needs investigations to discover the molecular mechanisms involved in the crisis. The disease of Baikal sponges is one of the processes resulting in the degradation of the littoral zone of the lake. Methods: The chloroplast genome fragment for the algae endosymbiont of Baikal sponge was assembled from metagenomic sequencing data. The distributions of polymorphic sites were obtained for the genome fragment, separately for samples from healthy sponge, diseased sponge and dead sponge tissues. Results: The comparative analysis of chloroplast genome sequences suggests that the symbiotic algae from Baikal sponge is close to Choricystis genus of unicellular algae. Also, the distributions of polymorphic sites allowed detection of the signs of extensive mutations in the chloroplasts isolated from the diseased sponge tissues. Conclusions: The study demonstrate the particular case of evolution at the molecular level due to the conditions of a severe crisis of a whole ecosystem in Lake Baikal. The detection of adaptive mutations in the chloroplast genome is an important feature which could represent the behavior of an ecosystem in the event of a severe crisis.

Mass disease and mortality of Baikal sponges

In recent years, signifcant changes in the ecological system of the coastal (littoral) zone, including mass death of the endemic representatives of the freshwater sponges of the Lubomirskiidae family, have been an urgent problem of Lake Baikal. Similar problems are known all over the world. Thus, mass disease and death of corals and sponges are indicated in the Mediterranean, Adriatic, Caribbean and other seas, which raises serious concerns about the future of these biocenoses (Olson et al., 2006; Webster, 2007; Wulff et al., 2007; Stabili et al., 2012). In Baikal, diseased sponges were frst found in 2011. The area of sponge disease is constantly expanding, and, to date, dying specimens have been found throughout the lake. The mass death of sponges occur in presence of the large-scale violation of the spatial distribution and the structure of phytocenoses in the littoral zone, but the causes of these phenomena are unknown. The relevance of the problem arises from the fact that changes in the littoral zone of Lake Baikal can signifcantly affect the productivity and composition of planktonic organisms and zoobenthos, which are the food base for fsh, as well as the quality of drinking water. The deterioration of the ecological state also affects the attractiveness of the lake for tourism. At the international level, serious intellectual and fnancial resources were mobilized to solve similar problems. Despite the obvious relevance, in Russia such studies are carried out irregularly by small groups of researchers.

Current state of the sponge fauna (Porifera: Lubomirskiidae) of Lake Baikal: Sponge disease and the problem of conservation of diversity

Abstract The spatial distribution of healthy and diseased sponges in Lake Baikal was investigated. The endemic sponges were studied in May − June 2015 at 11 transects within the depth range of 0 to 40 m. More than 500 sponge specimens were collected, and 29 taxa including 12 species and 17 morphospecies (sp. taxa) were identified. Among the 29 taxa, only B. martinsoni specimens and 14 morphospecies were healthy, whereas specimens of the other 11 species and 3 morphospecies were diseased, making up 35% of the total number of specimens (512). The different body forms of sponge found among the specimens were encrusting (66%), globulous (21%), and branched (13%). Sponges with necrosis signs, discolored specimens or covered with violet film or brown spots, were found in all transects under study (37 stations of 68). The percent cover of the bottom (per m2) by various body forms of sponges was evaluated with a sponge faunal state assessment test. Changes in the diversity and abundance of sponges due to their disease may jeopardize the functioning of the entire ecosystem of Lake Baikal which is a World Heritage Site. This baseline data on patterns of sponges, including their spatial distribution and percent cover, will be utilized to support future management and monitoring of Lake Baikal. Data on the current state of Baikal endemic sponge fauna are essential for understanding the causes and prognosis of disease processes of freshwater sponges.