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Major aggregations of nesting green turtles (Chelonia mydas) occur in the northern Red Sea, although little is known about the reproductive ecology of this endangered species in the region. To address this issue, we satellite-tracked 30 female green turtles to document their movements and to identify factors driving habitat use at two major rookeries in the Red Sea, Jazirat Mashabah (Mashabah Island) and Ras Al Baridi in Saudi Arabia. Between successive nesting events, turtles displayed high fidelity to nesting beaches and adjacent in-water habitats (inter-nesting habitats). Using generalized linear mixed models, we estimated the mean probability of nesting per beach emergence (nesting success rate) to be 0.628, and the mean duration between a successful nesting event and the successive emergence onto the beach (re-nesting interval) to be 10.8 days at each site. The nesting success rate was relatively high (>0.8) when the preceding daytime land surface temperature (LST) was lower than 37°C but decreased with elevated daytime LST (47°C). Re-nesting interval was longer at lower water temperatures and towards the end of the nesting season of individuals. Our study improves the robustness of abundance estimates from census data (e.g., track counts) and shows that the protection of nesting and inter-nesting habitats during a breeding season would be an effective conservation strategy for the species. We discuss how global warming could increase energy expenditure due to lowered nesting success, ultimately compromising the reproductive fitness of these populations.

Biofilms are a significant source of marine biofouling. Marine biofilm communities are established when microorganisms adhere to immersed surfaces. Despite the microbe-inhibiting effect of zinc surfaces, microbes can still attach to the surface and form biofilms. However, the diversity of biofilm-forming microbes that can attach to zinc surfaces and their common functional features remain elusive. Here, by analyzing 9,000,000 16S rRNA gene amplicon sequences and 270 Gb of metagenomic data, we comprehensively explored the taxa and functions related to biofilm formation in subtidal zones of the Red Sea. A clear difference was observed between the biofilm and adjacent seawater microbial communities in terms of the taxonomic structure at phylum and genus levels, and a huge number of genera were only present in the biofilms. Saturated alpha-diversity curves suggested the existence of more than 14,000 operational taxonomic units in one biofilm sample, which is much higher than previous estimates. Remarkably, the biofilms contained abundant and diverse transposase genes, which were localized along microbial chromosomal segments and co-existed with genes related to metal ion transport and resistance. Genomic analyses of two cyanobacterial strains that were abundant in the biofilms revealed a variety of metal ion transporters and transposases. Our analyses revealed the high diversity of biofilm-forming microbes that can attach to zinc surfaces and the ubiquitous role of transposase genes in microbial adaptation to toxic metal surfaces.

Mobile species often aggregate at predictable places and times to ensure that individuals find mates and breed in suitable habitats. Sea turtles demonstrate this life history trait, which can make these species highly susceptible to population declines if nesting habitats are lost or degraded. Conservation management thus requires knowledge of where and when turtles nest and changes in abundance in these habitats through time. Here, we compiled new and published data and used a novel analysis to describe seasonality, annual abundance and spatial distribution of nesting green (Chelonia mydas) and hawksbill (Eretmochelys imbricata) turtles in data-deficient populations that inhabit the Red Sea. Major new rookeries were identified for green turtles at Jazirat 1 Mashabah (113 and 179 nesting females in 2018 and 2019) and for hawksbill turtles at Jazirat Al Waqqadi (79 nesting females in 2018), both of which are located on nearshore islands of the Kingdom of Saudi Arabia in an area subject to industrial, residential and ecotourism developments. An upward trend in annual abundance of nesting sea turtles was estimated at some sites including Ras Al Baridi (Saudi Arabia), a major rookery of green turtles in the Red Sea, where the annual numbers increased from 14–110 individuals in 1982–1995 to 178 and 330 individuals in 2018 and 2019. This integrative work provides the most up-to-date, comprehensive information on nesting sea turtles in the Red Sea and documents a critical baseline for sea turtle conservation and future management effort.

Fish contamination has been extensively investigated along the Saudi coasts, but studies pertaining to bacterial pathogens are scarce. We conducted qualitative assessment and molecular identification of culture-dependent bacteria in 13 fish species from three coastal sites and a local fish market in Jeddah, Saudi Arabia. Bacterial counts of gills, skin, gut and muscle were examined on agar plates of Macconkey’s (Mac), Eosin Methylene Blue (EMB) and Thiosulfate Citrate Bile Salts (TCBS) culture media. Bacterial counts significantly differed between species, sources and feeding habits of examined fishes. Mugil cephalus exhibited higher counts on TCBS (all body parts), Mac (gills, muscle and gut) and EMB (gills and muscle). Fishes from Area I had higher bacterial loads, coinciding with those in seawater and sediment from the same site, indicating direct association between habitat conditions and the levels of bacterial contamination. By feeding habit, detritivorous fish harbored higher counts than herbivorous and carnivorous species. Bacterial counts of skin were higher in fish from market than field sites, and positively correlated with other body parts indicating the relation of surface bacterial load on the overall quality of fish. Rahnella aquatilis (Enterobacteriaceae) and Photobacterium damselae (Vibrionaceae) were among the dominant species from fish muscle based on 16S rRNA sequencing. These species are known human pathogens capable of causing foodborne illness with severe antibiotic resistance. Opportunistic pathogens, e.g. Hafnia sp. (Enterobacteriaceae) and Pseudomonas stutzeri (Pseudomonadaceae) also occurred in fish muscle. The inclusion of bacterial contamination in future monitoring efforts is thus crucial. Keywords: Marine environment, Fish, Bacterial contamination, Population count, Molecular identification, Sanger sequencing

All 209 congeners of polychlorinated biphenyls (PCBs) in edible fish tissue from the central Red Sea coast (Jeddah region) of Saudi Arabia were analyzed by isotope dilution high-resolution gas chromatography–mass spectrometry. The upper-bound total PCB (ΣPCB) levels in nine commonly consumed fish species from three areas were 0.2–82.5 ng/g wet weight (17–8450 ng/g lipid weight), which were at the lower end of reported global range and far below international tolerance limits (500–3000 ng/g ww). Dioxin-like congeners contributed up to 12.8% (mean 6.5%) to ΣPCB in tissue samples, with the total PCB toxic equivalencies (TEQs) at a tolerable range (0.05–2.6 pg TEQ/g ww or 2–238 pg TEQ/g lw) for all species. PCB profiles were dominated by moderately chlorinated homologs, mainly hexachlorobiphenyls, but less chlorinated congeners were also consistently elevated, notably in Siganus rivulatus (Area III) and Mugil cephalus (Area I). It remains to be ascertained if the latter were breakdown products or due to fresh inputs. The top congeners based on dominance by both occurrence and abundance were identified as potential markers of ΣPCB in fish tissue, which can be used for future selective biomonitoring in case of reasonable constraints on full congener approach.

We thank the Center for Environment and Water (CEW) at the Research Institute of King Fahd University of Petroleum and Minerals, Dhahran, Saudi Arabia, for providing research facilities and the Marine Studies Section team of CEW for performing the fieldwork and laboratory analyses. We express our sincere thanks to the General Authority of Meteorology and Environmental Protection, Saudi Arabia and Saudi Company for Environmental Works Ltd. for their constant support. We also thank M/S Al- Gosaibi for providing services of their Oceanographic Vessel.

This study assesses the presence of microplastic litter in the contents of the gastrointestinal tract of 26 commercial and non-commercial fish species from four difference habitats sampled along the Saudi Arabian coast of the Red Sea. A total of 178 individual were examined for microplastics. In total, 26 microplastic fragments were found. Of these, 16 being films (61.5%) and 10 being fishing thread (38.5%). FTIR analysis revealed that the most abundant polymers were polypropylene and polyethylene. Parascolosps eriomma species sampled at Jazan registered the highest number of ingested microplastic. This fish species is benthic and feeds on benthic invertebrates. Although differences in the abundance of microplastic ingestion among species were not statistically significant, a significant change was observed when the level of ingestion of microplastics particles was compared among the habitats. The higher abundance of microplastics particles may be related to the habitats of fish and the presence of microplastics debris near the seabed. The results of this study represent a first evidence that microplastic pollution represents an emerging threat to Red Sea fishes, their food web and human consumers.

The biology of biofilm in deep-sea environments is barely being explored. Here, biofilms were developed at the brine pool (characterized by limited carbon sources) and the normal bottom water adjacent to Thuwal cold seeps. Comparative metagenomics based on 50 Gb datasets identified polysaccharide degradation, nitrate reduction and proteolysis as enriched functional categories for brine biofilms. The genomes of two dominant species: a novel Deltaproteobacterium and a novel Epsilonproteobacterium in the brine biofilms were reconstructed. Despite rather small genome sizes, the Deltaproteobacterium possessed enhanced polysaccharide fermentation pathways, whereas the Epsilonproteobacterium was a versatile nitrogen reactor possessing nar, nap and nif gene clusters. These metabolic functions, together with specific regulatory and hypersaline-tolerant genes, made the two bacteria unique compared with their close relatives, including those from hydrothermal vents. Moreover, these functions were regulated by biofilm development, as both the abundance and the expression level of key functional genes were higher in later stage biofilms, and co-occurrences between the two dominant bacteria were demonstrated. Collectively, unique mechanisms were revealed: (i) polysaccharides fermentation, proteolysis interacted with nitrogen cycling to form a complex chain for energy generation, and (ii) remarkably exploiting and organizing niche-specific functions would be an important strategy for biofilm-dependent adaptation to the extreme conditions.

Fish are a healthful source of protein, but contaminants in some fish pose a risk. While there are multiple risk assessments from Europe and North America, there are far fewer for other parts of the world. We examined the risks from mercury, arsenic, lead, and other metals in fish consumed by people in Jeddah area, Saudi Arabia, using site-specific data on consumption patterns and metal levels in fish. The U.S. Environmental Protection Agency's Hazard Quotient (HQ) and cumulative Hazard Index (HI) for non-cancer endpoints and Carcinogenic Index for cancer were used to determine the health risk based on fish consumption rates. Of the 13 fish species examined, HQ was greater than 1 (indicating elevated risk) in two species for arsenic, and seven species for methylmercury. The cumulative HI for all metals was above 1 for all but three species of fish at the mean consumption rates. Generally, fish species with HI above 1 for one sampling location, had HI above 1 for all sampling locations. The implica...

Metal levels in fish have been extensively studied, but little data currently exists for the Middle East. We examined the levels of metals and metalloids (aluminum, arsenic, copper, manganese, selenium, zinc, and mercury) in the flesh of 13 fish species collected from three fishing sites and a local fish market in Jeddah, Saudi Arabia. We tested the following null hypotheses: (1) there are no interspecific differences in metal levels, (2) there are no differences in metal levels in fishes between market and fishing sites, (3) there are no size-related differences in metal levels, and (4) there are no differences in selenium:mercury molar ratio among different fish species. There were significant interspecific differences in concentrations for all metals. There was an order of magnitude difference in the levels of aluminum, arsenic, mercury, manganese, and selenium, indicating wide variation in potential effects on the fish themselves and on their predators. Fishes from Area II, close to a large commercial port, had the highest levels of arsenic, mercury, and selenium, followed by market fishes. Mercury was positively correlated with body size in 6 of the 13 fish species examined. Mercury was correlated positively with arsenic and selenium, but negatively with aluminum, cobalt, copper, manganese, and zinc. Selenium:mercury molar ratios varied significantly among species, with Carangoides bajad, Cephalopholis argus, Variola louti, and Ephinephelus tauvina having ratios below 10:1. These findings can be used in risk assessments, design of mercury reduction plans, development of fish advisories to protect public health, and future management decision-making.

Atlantis II and Discovery are two hydrothermal and hypersaline deep-sea pools in the Red Sea rift that are characterized by strong thermohalo-stratification and temperatures steadily peaking near the bottom. We conducted comprehensive vertical profiling of the microbial populations in both pools and highlighted the influential environmental factors. Pyrosequencing of the 16S rRNA genes revealed shifts in community structures vis-à-vis depth. High diversity and low abundance were features of the deepest convective layers despite the low cell density. Surprisingly, the brine interfaces had significantly higher cell counts than the overlying deep-sea water, yet they were lowest in diversity. Vertical stratification of the bacterial populations was apparent as we moved from the Alphaproteobacteria -dominated deep sea to the Planctomycetaceae - or Deferribacteres -dominated interfaces to the Gammaproteobacteria- dominated brine layers. Archaeal marine group I was dominant in the deep-sea water and interfaces, while several euryarchaeotic groups increased in the brine. Across sites, microbial phylotypes and abundances varied substantially in the brine interface of Discovery compared with Atlantis II, despite the near-identical populations in the overlying deep-sea waters. The lowest convective layers harbored interestingly similar microbial communities, even though temperature and heavy metal concentrations were very different. Multivariate analysis indicated that temperature and salinity were the major influences shaping the communities. The harsh conditions and the low-abundance phylotypes could explain the observed correlation in the brine pools.

A slow-growing, strictly aerobic, Gram-negative, coccus bacterial strain, designated KAUST100406-0324T, was isolated from sea-floor sediment collected from the Red Sea, Saudi Arabia. The catalase- and oxidase-positive strain was non-sporulating and only slightly halophilic. Optimum growth occurred at 20–25 °C and at pH values ranging from 7.0 to 8.0. The major cellular fatty acids of the strain were unsaturated C18 : 1ω6c and/or C18 : 1ω7c, C18 : 1ω7c 11-methyl and C16 : 1ω7c and/or C16 : 1ω6c. The major polar lipids were phosphatidylglycerol, phosphatidylethanolamine and two unidentified phospholipids. Ubiquinone 10 was the predominant lipoquinone. The DNA G+C content of strain KAUST100406-0324T was 64.0 mol%. Phylogenetic analysis of 16S rRNA gene sequences revealed that the novel strain belonged to the family Rhodobacteraceae of the class Alphaproteobacteria but formed a distinct evolutionary lineage from other bacterial species with validly published names. The 16S rRNA gene sequence of the novel strain was distantly related, but formed a monophyletic cluster with, those of bacteria from two moderately halophilic genera, Hwanghaeicola and Maribius . The similarity of the sequence between the novel strain KAUST100406-0324T and the type strains Hwanghaeicola aestuarii Y26T (accession number FJ230842), Maribius pelagius B5-6T (DQ514326) and Maribius salinus CL-SP27T (AY906863) were 94.5 %, 95.2 % and 95.3 %, respectively. Based on the physiological, phylogenetic and chemotaxonomic characteristics presented in this study, we propose that this strain represents a novel species of a new genus in the family Rhodobacteraceae , for which the name of Profundibacterium mesophilum gen. nov., sp. nov. was proposed, with KAUST100406-0324T ( = JCM 17872T = NRRL B-59665T) as the type strain.

A new cold brine seep system with microbial mats and metazoan assemblages was discovered by a remotely operated vehicle (ROV) on the Saudi continental margin of central Red Sea. Now named as Thuwal Seeps, it has a shallow brine pool between 840 and 850 m water depths that is formed by focused brine expulsions from two sites (Seep I: 22°17.3′N, 38°53.8′E; Seep II: 22°16.9′N, 38°53.9′E). The seep is located at the base of a steep wall rock closer to the shore (20 km) than to the axial trough (120 km). The brine pool does not exhibit a significant thermal anomaly (

The anti-neoplastic agent didemnin B from the Caribbean tunicate Trididemnum solidum was the first marine drug to be clinically tested in humans. Because of its limited supply and its complex cyclic depsipeptide structure, considerable challenges were encountered during didemnin B's development that continue to limit aplidine (dehydrodidemnin B), which is currently being evaluated in numerous clinical trials. Herein we show that the didemnins are bacterial products produced by the marine α-proteobacteria Tistrella mobilis and Tistrella bauzanensis via a unique post-assembly line maturation process. Complete genome sequence analysis of the 6,513,401 bp T. mobilis strain KA081020-065 with its five circular replicons revealed the putative didemnin biosynthetic gene cluster (did) on the 1,126,962 bp megaplasmid pTM3. The did locus encodes a 13-module hybrid non-ribosomal peptide synthetase-polyketide synthase enzyme complex organized in a collinear arrangement for the synthesis of the fatty acylglutamine ester derivatives didemnins X and Y rather than didemnin B as first anticipated. Imaging mass spectrometry of T. mobilis bacterial colonies captured the time-dependent extracellular conversion of the didemnin X and Y precursors to didemnin B, in support of an unusual post-synthetase activation mechanism. Significantly, the discovery of the didemnin biosynthetic gene cluster may provide a long-term solution to the supply problem that presently hinders this group of marine natural products and pave the way for the genetic engineering of new didemnin congeners.

The 1991 Gulf oil spill heavily impacted the coastal areas of the Saudi waters of the Arabian Gulf and recent studies have indicated that even 15 years after the incident, macrobenthos had not completely recovered in the sheltered bays in the affected region such as, Manifa Bay. This study investigates the community conditions of macrobenthos in the open waters in one of the impacted areas, Al-Khafji waters, about 14 years after the spill. Diversity measures and community structure analyses indicate a healthy status of polychaete communities. The BOPA index reveals that oil sensitive amphipods were recolonized in the study area. This confirms that the benthic communities of the oil spill impacted area had taken only

Shallow water bays located in the western Arabian Gulf experience harsh environmental conditions. Some of these bays, including Manifa-Tanajib Bay System (MTBS), were also exposed to the 1991 oil pollution event. This study investigates the status of the macrobenthos in MTBS during 2006. This bay system is characterized by very shallow inner bays with elevated salinity and temperature compared to the rest of the bay area. As a result mainly of the hyper salinity, the inner bay communities are distinct from the outer bay communities. Overall, fairly high species richness with several rare species was observed. High Shannon-Wiener diversity values and ABC plots indicated the healthy status of the polychaete communities, while BOPA index indicated slightly polluted status in 20% of the stations. The oil sensitive amphipods were not completely re-colonized in 20% of the stations, even after 15 years of recovery from the 1991 oil spill.

Hydrothermal ecosystems have a wide distribution on Earth and many can be found in the basin of the Red Sea. Production of aromatic compounds occurs in a temperature window of ∼60–150 °C by utilizing organic debris. In the past 50 years, the temperature of the Atlantis II Deep brine pool in the Red Sea has increased from 56 to 68 °C, whereas the temperature at the nearby Discovery Deep brine pool has remained relatively stable at about 44 °C. In this report, we confirmed the presence of aromatic compounds in the Atlantis II brine pool as expected. The presence of the aromatic compounds might have disturbed the microbes in the Atlantis II. To show shifted microbial communities and their metabolisms, we sequenced the metagenomes of the microbes from both brine pools. Classification based on metareads and the 16S rRNA gene sequences from clones showed a strong divergence of dominant bacterial species between the pools. Bacteria capable of aromatic degradation were present in the Atlantis II brine pool. A comparison of the metabolic pathways showed that several aromatic degradation pathways were significantly enriched in the Atlantis II brine pool, suggesting the presence of aromatic compounds. Pathways utilizing metabolites derived from aromatic degradation were also significantly affected. In the Discovery brine pool, the most abundant genes from the microbes were related to sugar metabolism pathways and DNA synthesis and repair, suggesting a different strategy for the utilization of carbon and energy sources between the Discovery brine pool and the Atlantis II brine pool.

Marine sponges are associated with a remarkable array of microorganisms. Using a tag pyrosequencing technology, this study was the first to investigate in depth the microbial communities associated with three Red Sea sponges, Hyrtios erectus, Stylissa carteri and Xestospongia testudinaria. We revealed highly diverse sponge-associated bacterial communities with up to 1000 microbial operational taxonomic units (OTUs) and richness estimates of up to 2000 species. Altogether, 26 bacterial phyla were detected from the Red Sea sponges, 11 of which were absent from the surrounding sea water and 4 were recorded in sponges for the first time. Up to 100 OTUs with richness estimates of up to 300 archaeal species were revealed from a single sponge species. This is by far the highest archaeal diversity ever recorded for sponges. A non-negligible proportion of unclassified reads was observed in sponges. Our results demonstrated that the sponge-associated microbial communities remained highly consistent in the same sponge species from different locations, although they varied at different degrees among different sponge species. A significant proportion of the tag sequences from the sponges could be assigned to one of the sponge-specific clusters previously defined. In addition, the sponge-associated microbial communities were consistently divergent from those present in the surrounding sea water. Our results suggest that the Red Sea sponges possess highly sponge-specific or even sponge-species-specific microbial communities that are resistant to environmental disturbance, and much of their microbial diversity remains to be explored.

Sponge diseases have been widely reported, yet the causal factors and major pathogenic microbes remain elusive. In this study, two individuals of the sponge Crella cyathophora in total that showed similar disease-like characteristics were collected from two different locations along the Red Sea coast separated by more than 30 kilometers. The disease-like parts of the two individuals were both covered by green surfaces, and the body size was much smaller compared with adjacent healthy regions. Here, using high-throughput pyrosequencing technology, we investigated the prokaryotic communities in healthy and disease-like sponge tissues as well as adjacent seawater. Microbes in healthy tissues belonged mainly to the Proteobacteria, Cyanobacteria and Bacteroidetes, and were much more diverse at the phylum level than reported previously. Interestingly, the disease-like tissues from the two sponge individuals underwent shifts of prokaryotic communities and were both enriched with a novel clade affiliated with the phylum Verrucomicrobia, implying its intimate connection with the disease-like Red Sea sponge C. cyathophora. Enrichment of the phylum Verrucomicrobia was also considered to be correlated with the presence of algae assemblages forming the green surface of the disease-like sponge tissues. This finding represents an interesting case of sponge disease and is valuable for further study.

The functional basis for species sorting theory remains elusive, especially for microbial community assembly in deep-sea environments. Using artificial surface-based biofilm models, our recent work revealed taxonomic succession during biofilm development in a newly defined cold seep system, the Thuwal cold seeps II, which comprises a brine pool and the adjacent normal bottom water (NBW) to form a metacommunity via the potential immigration of organisms from one patch to another. Here, we designed an experiment to investigate the effects of environmental switching between the brine pool and the NBW on biofilm assembly, which could reflect environmental filtering effects during bacterial immigration to new environments. Analyses of 16S rRNA genes of 71 biofilm samples suggested that the microbial composition of biofilms established in new environments was determined by both the source community and the incubation conditions. Moreover, a comparison of 18 metagenomes provided evidence for biofilm community assembly that was based primarily on functional features rather than taxonomic identities; metal ion resistance and amino acid metabolism were the major species sorting determinants for the succession of biofilm communities. Genome binning and pathway reconstruction of two bacterial species (Marinobacter sp. and Oleispira sp.) further demonstrated metal ion resistance and amino acid metabolism as functional traits conferring the survival of habitat generalists in both the brine pool and NBW. The results of this study shed new light on microbial community assembly in special habitats and bridge a gap in species sorting theory.

In deep-sea geothermal rift zones, the dispersal of hydrothermal fluids of moderately-high temperatures typically forms subseafloor mounds. Major mineral components of the crust covering the mound are barite and metal sulfides. As a result of the continental rifting along the Red Sea, metalliferous sediments accumulate on the seafloor of the Atlantis II Deep. In the present study, a barite crust was identified in a sediment core from the Atlantis II Deep, indicating the formation of a hydrothermal mound at the sampling site. Here, we examined how such a dense barite crust could affect the local environment and the distribution of microbial inhabitants. Our results demonstrate distinctive features of mineral components and microbial communities in the sediment layers separated by the barite crust. Within the mound, archaea accounted for 65% of the community. In contrast, the sediments above the barite boundary were overwhelmed by bacteria. The composition of microbial communities under the mound was similar to that in the sediments of the nearby Discovery Deep and marine cold seeps. This work reveals the zonation of microbial communities after the formation of the hydrothermal mound in the subsurface sediments of the rift basin.

Studies focusing on biofilm assembly in deep-sea environments are rarely conducted. To examine the effects of substrate type on microbial community assembly, biofilms were developed on different substrates for different durations at two locations in the Red Sea: in a brine pool and in nearby bottom water (NBW) adjacent to the Thuwal cold seep II. The composition of the microbial communities in 51 biofilms and water samples were revealed by classification of pyrosequenced 16S rRNA gene amplicons. Together with the microscopic characteristics of the biofilms, the results indicate a stronger selection effect by the substrates on the microbial assembly in the brine pool compared with the NBW. Moreover, the selection effect by substrate type was stronger in the early stages compared with the later stages of the biofilm development. These results are consistent with the hypotheses proposed in the framework of species sorting theory, which states that the power of species sorting during microbial community assembly is dictated by habitat conditions, duration and the structure of the source community. Therefore, the results of this study shed light on the control strategy underlying biofilm-associated marine fouling and provide supporting evidence for ecological theories important for understanding the formation of deep-sea biofilms.

“Candidatus Synechococcus spongiarum” is a cyanobacterial symbiont widely distributed in sponges, but its functions at the genome level remain unknown. Here, we obtained the draft genome (1.66 Mbp, 90% estimated genome recovery) of “Ca. Synechococcus spongiarum” strain SH4 inhabiting the Red Sea sponge Carteriospongia foliascens. Phylogenomic analysis revealed a high dissimilarity between SH4 and free-living cyanobacterial strains. Essential functions, such as photosynthesis, the citric acid cycle, and DNA replication, were detected in SH4. Eukaryoticlike domains that play important roles in sponge-symbiont interactions were identified exclusively in the symbiont. However, SH4 could not biosynthesize methionine and polyamines and had lost partial genes encoding low-molecular-weight peptides of the photosynthesis complex, antioxidant enzymes, DNA repair enzymes, and proteins involved in resistance to environmental toxins and in biosynthesis of capsular and extracellular polysaccharides. These genetic modifications imply that “Ca. Synechococcus spongiarum” SH4 represents a low-light-adapted cyanobacterial symbiont and has undergone genome streamlining to adapt to the sponge’s mild intercellular environment., IMPORTANCE Although the diversity of sponge-associated microbes has been widely studied, genome-level research on sponge symbionts and their symbiotic mechanisms is rare because they are unculturable. “Candidatus Synechococcus spongiarum” is a widely distributed uncultivated cyanobacterial sponge symbiont. The genome of this symbiont will help to characterize its evolutionary relationship and functional dissimilarity to closely related free-living cyanobacterial strains. Knowledge of its adaptive mechanism to the sponge host also depends on the genome-level research. The data presented here provided an alternative strategy to obtain the draft genome of “Ca. Synechococcus spongiarum” strain SH4 and provide insight into its evolutionary and functional features.

A hypoxic/suboxic brine pool at a depth of about 850 m was discovered near the Thuwal cold seeps in the Red Sea. Filled with high concentrations of hydrogen sulfide and ammonia, such a brine pool might limit the spread of eukaryotic organisms. Here, we compared the communities of the eukaryotic microbes in a microbial mat, sediments and water samples distributed in 7 sites within and adjacent to the brine pool. Taxonomic classification of the pyrosequenced 18S rRNA amplicon reads showed that fungi highly similar to the species identified along the Arabic coast were almost ubiquitous in the water and sediment samples, supporting their wide distribution in various environments. The microbial mat displayed the highest species diversity and contained grazers and a considerable percentage of unclassified species. Phylogeny-based methods revealed novel lineages representing a majority of the reads from the interface between the sea water and brine pool. Phylogenetic relationships with more reference sequences suggest that the lineages were affiliated with novel Alveolata and Euglenozoa inhabiting the interface where chemosynthetic prokaryotes are highly proliferative due to the strong chemocline and halocline. The brine sediments harbored abundant species highly similar to invertebrate gregarine parasites identified in different oxygen-depleted sediments. Therefore, the present findings support the uniqueness of some microbial eukaryotic groups in this cold seep brine system.

Using microscopic and molecular techniques combined with computational analysis, this study examined the structure and composition of microbial communities in biofilms that formed on different artificial substrates in a brine pool and on a seep vent of a cold seep in the Red Sea to test our hypothesis that initiation of the biofilm formation and spreading mode of microbial structures differs between the cold seep and the other aquatic environments. Biofilms on different substrates at two deployment sites differed morphologically, with the vent biofilms having higher microbial abundance and better structural features than the pool biofilms. Microbes in the pool biofilms were more taxonomically diverse and mainly composed of various sulfate-reducing bacteria whereas the vent biofilms were exclusively dominated by sulfur-oxidizing Thiomicrospira. These results suggest that the redox environments at the deployment sites might have exerted a strong selection on microbes in the biofilms at two sites whereas the types of substrates had limited effects on the biofilm development.

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.

It is usually assumed that metabolic constraints restrict deep-sea corals to cold-water habitats, with ‘deep-sea’ and ‘cold-water’ corals often used as synonymous. Here we report on the first measurements of biological characters of deep-sea corals from the central Red Sea, where they occur at temperatures exceeding 20°C in highly oligotrophic and oxygen-limited waters. Low respiration rates, low calcification rates and minimized tissue cover indicate that a reduced metabolism is one of the key adaptations to prevailing environmental conditions. We investigated four sites and encountered six species of which at least two appear to be undescribed. One species is previously reported from the Red Sea but occurs in deep cold waters outside the Red Sea raising interesting questions about presumed environmental constraints for other deep-sea corals. Our findings suggest that the present understanding of deep-sea coral persistence and resilience needs to be revisited.

In the Red Sea, two neighboring deep-sea brine pools, Atlantis II and Discovery, have been studied extensively, and the results have shown that the temperature and concentrations of metal and methane in Atlantis II have increased over the past decades. Therefore, we investigated changes in the microbial community and metabolic pathways. Here, we compared the metagenomes of the two pools to each other and to those of deep-sea water samples. Archaea were generally absent in the Atlantis II metagenome; Bacteria in the metagenome were typically heterotrophic and depended on aromatic compounds and other extracellular organic carbon compounds as indicated by enrichment of the related metabolic pathways. In contrast, autotrophic Archaea capable of CO2 fixation and methane oxidation were identified in Discovery but not in Atlantis II. Our results suggest that hydrothermal conditions and metal precipitation in the Atlantis II pool have resulted in elimination of the autotrophic community and methanogens.

Microbial associations with corals are common and are most likely symbiotic, although their diversity and relationships with environmental factors and host species remain unclear. In this study, we adopted a 16S rRNA gene tag-pyrosequencing technique to investigate the bacterial communities associated with three stony Scleractinea and two soft Octocorallia corals from three locations in the Red Sea. Our results revealed highly diverse bacterial communities in the Red Sea corals, with more than 600 ribotypes detected and up to 1,000 species estimated from a single coral species. Altogether, 21 bacterial phyla were recovered from the corals, of which Gammaproteobacteria was the most dominant group, and Chloroflexi , Chlamydiae , and the candidate phylum WS3 were reported in corals for the first time. The associated bacterial communities varied greatly with location, where environmental conditions differed significantly. Corals from disturbed areas appeared to share more similar bacterial communities, but larger variations in community structures were observed between different coral species from pristine waters. Ordination methods identified salinity and depth as the most influential parameters affecting the abundance of Vibrio , Pseudoalteromonas , Serratia , Stenotrophomonas , Pseudomonas , and Achromobacter in the corals. On the other hand, bacteria such as Chloracidobacterium and Endozoicomonas were more sensitive to the coral species, suggesting that the host species type may be influential in the associated bacterial community, as well. The combined influences of the coral host and environmental factors on the associated microbial communities are discussed. This study represents the first comparative study using tag-pyrosequencing technology to investigate the bacterial communities in Red Sea corals.

The complexity and dynamics of microbial metagenomes may be evaluated by genome size, gene duplication and the disruption rate between lineages. In this study, we pyrosequenced the metagenomes of microbes obtained from the brine and sediment of a deep-sea brine pool in the Red Sea to explore the possible genomic adaptations of the microbes in response to environmental changes. The microbes from the brine and sediments (both surface and deep layers) of the Atlantis II Deep brine pool had similar communities whereas the effective genome size varied from 7.4 Mb in the brine to more than 9 Mb in the sediment. This genome expansion in the sediment samples was due to gene duplication as evidenced by enrichment of the homologs. The duplicated genes were highly disrupted, on average by 47.6% and 70% for the surface and deep layers of the Atlantis II Deep sediment samples, respectively. The disruptive effects appeared to be mainly due to point mutations and frameshifts. In contrast, the homologs from the Atlantis II Deep brine sample were highly conserved and they maintained relatively small copy numbers. Likely, the adaptation of the microbes in the sediments was coupled with pseudogenizations and possibly functional diversifications of the paralogs in the expanded genomes. The maintenance of the pseudogenes in the large genomes is discussed.

The ecosystems of the Red Sea are among the least-explored microbial habitats in the marine environment. In this study, we investigated the microbial communities in the water column overlying the Atlantis II Deep and Discovery Deep in the Red Sea. Taxonomic classification of pyrosequencing reads of the 16S rRNA gene amplicons showed vertical stratification of microbial diversity from the surface water to 1500 m below the surface. Significant differences in both bacterial and archaeal diversity were observed in the upper (20 [corrected] and 50 m) and deeper layers (200 and 1500 m). There were no obvious differences in community structure at the same depth for the two sampling stations. The bacterial community in the upper layer was dominated by Cyanobacteria whereas the deeper layer harbored a large proportion of Proteobacteria. Among Archaea, Euryarchaeota, especially Halobacteriales, were dominant in the upper layer but diminished drastically in the deeper layer where Desulfurococcales belonging to Crenarchaeota became the dominant group. The results of our study indicate that the microbial communities sampled in this study are different from those identified in water column in other parts of the world. The depth-wise compositional variation in the microbial communities is attributable to their adaptations to the various environments in the Red Sea.