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Antibiotic resistance (AR) is an alarming global health concern, causing an annual death rate of more than 35,000 deaths in the US. AR is a natural phenomenon, reported in several pristine environments. In this study, we report AR in pristine Red Sea deep brine pools. Antimicrobial resistance genes (ARGs) were detected for several drug classes with tetracycline and macrolide resistance being the most abundant. As expected, ARGs abundance increased in accordance with the level of human impact with pristine Red Sea samples having the lowest mean ARG level followed by estuary samples, while activated sludge samples showed a significantly higher ARG level. ARG hierarchical clustering grouped drug classes for which resistance was detected in Atlantis II Deep brine pool independent of the rest of the samples. ARG abundance was significantly lower in the Discovery Deep brine pool. A correlation between integrons and ARGs abundance in brine pristine samples could be detected, while insertion sequences and plasmids showed a correlation with ARGs abundance in human-impacted samples not seen in brine pristine samples. This suggests different roles of distinct mobile genetic elements (MGEs) in ARG distribution in pristine versus human-impacted sites. Additionally, we showed the presence of mobile antibiotic resistance genes in the Atlantis II brine pool as evidenced by the co-existence of integrases and plasmid replication proteins on the same contigs harboring predicted multidrug-resistant efflux pumps. This study addresses the role of non-pathogenic environmental bacteria as a silent reservoir for ARGs, and the possible horizontal gene transfer mechanism mediating ARG acquisition. [ABSTRACT FROM AUTHOR]

Deep-sea anoxic brine pools are unique and extreme, yet habitable environments. However, their extent and processes of formation are not fully understood. Using geophysical analysis and seafloor surveying, we discovered the eastmost brine pools known in the ultraoligotrophic Eastern Mediterranean Sea, at the Palmahim Disturbance offshore Israel (~1150 m water depth). These brine pools are located directly above a ~1km wide piece of the Messinian evaporites section, which was up thrusted to ~350 m below the seafloor. We sampled brines and short cores to characterize the chemical composition of several small (up to 5m diameter) anoxic, methanic and warm (21.6°C) brine pools and adjacent seafloor sediments porewater. The maximal salinities measured at the pools and adjacent porewater were 63.9 and 72 PSU, respectively. The brines are characterized by enriched Na and Cl concentrations by a factor of ~1.8 and depleted Mg, SO4, K and Ca contents by factors of circa 6, 3, 2 and ~1.3, respectively, compared to the ambient seawater. Relations of the major element concentrations reveal a mixing curve between seawater and enriched Na/Cl and depleted Mg/Cl, K/Cl and SO4/Cl end-members, and do not coincide with relics of fossil residual evaporated seawater. We propose their composition reflects: 1) dissolution of Messinian halite (NaCl) by seawater, supported by their low Br/Cl ratios; 2) additional small rise in Na/Cl ratios due to the impact of clay mineral dehydration or/and dissolution of trace (~1% of the Na) amounts of detrital trona (Na3H(CO3)2•2H20), coinciding with the enriched alkalinity concentrations; 3) diagenesis processes depleting Mg, K and SO4, mainly by the formation of authigenic K-rich Mg-smectite, clay mineral dehydration, dolomitization/Mg-calcite precipitation and redox processes. The δ18O and δD values of the Palmahim brine may reflect the impact of clay mineral dehydration. Comparison to all other East Mediterranean brine lakes shows that the Palmahim brine pool system represents similar provenance of brines as observed for the Eastern Mediterranean Napoli, Nadir and Tyro lakes, while potentially recording additional processes attributed to its proximity to the coastal area.

The Red Sea is a marine environment with unique chemical characteristics and physical topographies. Among the various habitats offered by the Red Sea, the deep-sea brine pools are the most extreme in terms of salinity, temperature and metal contents. Nonetheless, the brine pools host rich polyextremophilic bacterial and archaeal communities. These microbial communities are promising sources for various classes of enzymes adapted to harsh environments – extremozymes. Extremozymes are emerging as novel biocatalysts for biotechnological applications due to their ability to perform catalytic reactions under harsh biophysical conditions, such as those used in many industrial processes. In this review, we provide an overview of the extremozymes from different Red Sea brine pools and discuss the overall biotechnological potential of the Red Sea proteome.

The Red Sea is a marine environment with unique chemical characteristics and physical topographies. Among the various habitats offered by the Red Sea, the deep-sea brine pools are the most extreme in terms of salinity, temperature and metal contents. Nonetheless, the brine pools host rich polyextremophilic bacterial and archaeal communities. These microbial communities are promising sources for various classes of enzymes adapted to harsh environments – extremozymes. Extremozymes are emerging as novel biocatalysts for biotechnological applications due to their ability to perform catalytic reactions under harsh biophysical conditions, such as those used in many industrial processes. In this review, we provide an overview of the extremozymes from different Red Sea brine pools and discuss the overall biotechnological potential of the Red Sea proteome. [ABSTRACT FROM AUTHOR]

Abstract Background Microbial species in the brine pools of the Red Sea and the brine pool-seawater interfaces are exposed to high temperature, high salinity, low oxygen levels and high concentrations of heavy metals. As adaptations to these harsh conditions require a large suite of secondary metabolites, these microbes have a huge potential as a source of novel anticancer molecules. Methods A total of 60 ethyl-acetate extracts of newly isolated strains from extreme environments of the Red-Sea were isolated and tested against several human cancer cell lines for potential cytotoxic and apoptotic activities. Results Isolates from the Erba brine-pool accounted for 50% of active bacterial extracts capable of inducing 30% or greater inhibition of cell growth. Among the 60 extracts screened, seven showed selectivity towards triple negative BT20 cells compared to normal fibroblasts. Conclusion In this study, we identified several extracts able to induce caspase-dependent apoptosis in various cancer cell lines. Further investigations and isolation of the active compounds of these Red Sea brine pool microbes may offer a chemotherapeutic potential for cancers with limited treatment options.

A quantitative analysis of coccoliths is presented in 18 core-top samples ranging between 26° N and 21° N and covering two major deep brine pools in the northern part of the Red Sea. Non-brine sites are characterized by rich coccoliths that may reach up to 3.31 × 109 coccoliths/g made by 22 species, whereas brine sites of Shaban and Kebrit Deeps with additional two non-brine sites are characterized by a decline in coccoliths/g (3.25 × 108 coccoliths/g), Shannon diversity, CaCO3 (%), and high TOC (%). Carbonate dissolution, inferred by qualitative observation and quantitative indices, was only observed at one brine site GeoB7828 in Kebrit Deep. This suggests that the decline in coccolith assemblages may not entirely be attributed to carbonate dissolution. The major decline, however, is probably related to the suspension of fecal pellets and marine aggregates containing delicate coccolith shields within a nepheloid layer and subsequently grazed by zooplankters in which reduced the numbers of coccolith that reached the bottom of the brine sites, or alternatively a deep-sea flow current that carried and remobilized some suspended particles outside the brine pool. Latitudinal fluctuations of eutrophic/oligotrophic coccoliths suggest profound trophic changes in the photic zone in the northern part of the Red Sea. C. braarudii, a valuable nutrient-indicator species is here reported for the first time, along with G. oceanica, H. carteri as well as biogenic opal dominating the assemblage between 26° N and 24° N, suggesting elevated nutrient conditions and supporting recent high chl-a records, whereas areas between 21° N and 23° N lie under oligotrophic conditions due to the presence of U. sibogae, U. tenuis, R. clavigera, F. profunda, and S. pulchra. [ABSTRACT FROM AUTHOR]

The lower convective layer (LCL) at Atlantis II brine pool of the Red sea represents one of the exceptional, unique ecosystems. Thioredoxin is a multi-functional antioxidant redox protein that has a crucial role in various vital cellular processes. In the current study, a functional metagenomics approach was used to isolate and characterize thioredoxin from the LCL of Atlantis II Deep brine pool (Trx-ATII). From the metagenomic DNA of the LCL, the thioredoxin gene was directly retrieved and sequenced. Sequence analysis showed that the gene belonged to thioredoxin-like superfamily with classical Trx motif (-CXXC-). Phylogenetic analysis revealed that Trx-ATII was closely related to Trx of Prochlorococcus marinus with a maximum identity of 86%. Successfully, Trx-ATII was cloned and expressed in E. coli , where the purified protein had M.wt of 16 kDa. Characterization studies revealed that Trx-ATII protein is halophilic; can tolerate up to 2.5 M NaCl and thermostable, where 90% of its activity was retained at 60 °C. Trx-ATII can reduce both DTNB and insulin disulfide- containing substrates. In conclusion, a unique thioredoxin protein was isolated from a harsh environment that can maintain its activity under extreme conditions of salinity and temperature as a promising redox protein for biotechnological applications. [ABSTRACT FROM AUTHOR]

The adaptation of sporeformers to extreme environmental conditions is frequently questioned due to their capacity to produce highly resistant endospores that are considered as resting contaminants, not representing populations adapted to the system. In this work, in order to gain a better understanding of bacterial adaptation to extreme habitats, we investigated the phenotypic and genomic characteristics of the halophile Virgibacillus sp. 21D isolated from the seawater-brine interface (SBI) of the MgCl2-saturated deep hypersaline anoxic basin Discovery located in the Eastern Mediterranean Sea. Vegetative cells of strain 21D showed the ability to grow in the presence of high concentrations of MgCl2, such as 14.28% corresponding to 1.5 M. Biolog phenotype MicroArray (PM) was adopted to investigate the strain phenotype, with reference to carbon energy utilization and osmotic tolerance. The strain was able to metabolize only 8.4% of 190 carbon sources provided in the PM1 and PM2 plates, mainly carbohydrates, in accordance with the low availability of nutrients in its habitat of origin. By using in silico DNA-DNA hybridization the analysis of strain 21D genome, assembled in one circular contig, revealed that the strain belongs to the species Virgibacillus dokdonensis. The genome presented compatible solute-based osmoadaptation traits, including genes encoding for osmotically activated glycine-betaine/carnitine/choline ABC transporters, as well as ectoine synthase enzymes. Osmoadaptation of the strain was then confirmed with phenotypic assays by using the osmolyte PM9 Biolog plate and growth experiments. Furthermore, the neutral isoelectric point of the reconstructed proteome suggested that the strain osmoadaptation was mainly mediated by compatible solutes. The presence of genes involved in iron acquisition and metabolism indicated that osmoadaptation was tailored to the iron-depleted saline waters of the Discovery SBI. Overall, both phenomics and genomics highlighted the potential capability of V. dokdonensis 21D vegetative cells to adapt to the environmental conditions in Discovery SBI.

The adaptation of sporeformers to extreme environmental conditions is frequently questioned due to their capacity to produce highly resistant endospores that are considered as resting contaminants, not representing populations adapted to the system. In this work, in order to gain a better understanding of bacterial adaptation to extreme habitats, we investigated the phenotypic and genomic characteristics of the halophile Virgibacillus sp. 21D isolated from the seawater-brine interface (SBI) of the MgCl2-saturated deep hypersaline anoxic basin Discovery located in the Eastern Mediterranean Sea. Vegetative cells of strain 21D showed the ability to grow in the presence of high concentrations of MgCl2, such as 14.28% corresponding to 1.5 M. Biolog phenotype MicroArray (PM) was adopted to investigate the strain phenotype, with reference to carbon energy utilization and osmotic tolerance. The strain was able to metabolize only 8.4% of 190 carbon sources provided in the PM1 and PM2 plates, mainly carbohydrates, in accordance with the low availability of nutrients in its habitat of origin. By using in silico DNA-DNA hybridization the analysis of strain 21D genome, assembled in one circular contig, revealed that the strain belongs to the species Virgibacillus dokdonensis. The genome presented compatible solute-based osmoadaptation traits, including genes encoding for osmotically activated glycine-betaine/carnitine/choline ABC transporters, as well as ectoine synthase enzymes. Osmoadaptation of the strain was then confirmed with phenotypic assays by using the osmolyte PM9 Biolog plate and growth experiments. Furthermore, the neutral isoelectric point of the reconstructed proteome suggested that the strain osmoadaptation was mainly mediated by compatible solutes. The presence of genes involved in iron acquisition and metabolism indicated that osmoadaptation was tailored to the iron-depleted saline waters of the Discovery SBI. Overall, both phenomics and genomics highlighted the potential capability of V. dokdonensis 21D vegetative cells to adapt to the environmental conditions in Discovery SBI. [ABSTRACT FROM AUTHOR]

The lower convective layer (LCL) of the Atlantis II brine pool of the Red sea is a unique environment in terms of high salinity, temperature, and high concentrations of heavy metals. Mercuric reductase enzymes functional in such extreme conditions could be considered as a potential tool in the environmental detoxification of mercurial poisoning and might alleviate ecological hazards in the mining industry. Here, we constructed a mercuric reductase library from Atlantis II, from which we identified genes encoding two thermostable mercuric reductase (MerA) isoforms: one is halophilic (designated ATII-LCL) while the other is not (designated ATII-LCL-NH). The ATII-LCL MerA has a short motif composed of four aspartic acids (4D414-417) and two characteristic signature boxes that played a crucial role in its thermal stability. To further understand the mechanism behind the thermostability of the two studied enzymes, we mutated the isoform ATII-LCL-NH and found that the substitution of 2D at positions 415/416 enhanced the thermal stability, while other mutations had the opposite effect. The 2D mutant showed superior thermal tolerance as it retained 81% of its activity after 10 minutes incubation at 70°C. Three-dimensional structure prediction revealed newly formed salt-bridges and H-bonds in the 2D mutant, compared to the parent molecule. To the best of our knowledge, this study is the first to rationally design a mercuric reductase with enhanced thermal stability, which we propose to have a strong potential in bioremediation of mercurial poisoning. [ABSTRACT FROM AUTHOR]

Prediction of the geometry and petrophysical properties of dolomite geobodies depends on understanding both the hydrological system supplying reactive fluids and the chemistry of these fluids. However, patterns are complicated by the non-linear response of the diagenetic system to depositional texture, which controls both fluid flux via permeability architecture and reaction rate via effective surface area. This study explores interactions between extrinsic controls (spatial distribution of brine composition and temperature) and intrinsic controls (permeability and reactivity) using local and regional scale reactive transport models of sequential episodes of brine reflux that resulted in partial dolomitisation of the Mississippian Madison ramp. Inter-well scale models show preferential early dolomitisation of fine grained, more reactive beds. Pervasive dolomitisation can occur most readily beneath the brine pool where flow is perpendicular to bedding, and is most rapid at high brine fluxes. Down-dip of the brine pool, bedding-parallel flow is focused in relatively permeable coarse grained beds, providing reactants for strongly preferential alteration of intervening more reactive fine grained beds. In contrast, thicker sequences of fine grained beds dolomitise more slowly, limited by the rate of supply of magnesium. Regional-scale models, with injection of brines of increasing salinity towards the ramp interior, reproduce the observed pattern of dolomitisation. However, more realistic simulations in which reflux is driven by lateral density contrasts, generate flow rates orders of magnitude too low for significant dolomitisation. Simulations suggest pervasive dolomitisation of epeiric-scale ramps by a platform-wide reflux circulation, as often invoked, is not feasible. Rather, dolomitisation of such extensive systems critically requires local-scale flow systems, such as may result from topographically-controlled variations in restriction of platform-top seawater circulation. [ABSTRACT FROM AUTHOR]

Oceanic deep hypersaline anoxic basins (DHABs) are characterized by drastic changes in physico-chemical conditions in the transition from overlaying seawater to brine body. Brine-seawater interfaces (BSIs) of several DHABs across the Mediterranean Sea have been shown to possess methanogenic and sulfate-reducing activities, yet no systematic studies have been conducted to address the potential functional diversity of methanogenic and sulfate-reducing communities in the Red Sea DHABs. Here, we evaluated the relative abundance of Bacteria and Archaea using quantitative PCR and conducted phylogenetic analyses of nearly full-length 16S rRNA genes as well as functional marker genes encoding the alpha subunits of methyl-coenzyme M reductase ( mcrA ) and dissimilatory sulfite reductase ( dsrA ). Bacteria predominated over Archaea in most locations, the majority of which were affiliated with Deltaproteobacteria , while Thaumarchaeota were the most prevalent Archaea in all sampled locations. The upper convective layers of Atlantis II Deep, which bear increasingly harsh environmental conditions, were dominated by members of the class Thermoplasmata (Marine Benthic Group E and Mediterranean Sea Brine Lakes Group 1). Our study revealed unique microbial compositions, the presence of niche-specific groups, and collectively, a higher diversity of sulfate-reducing communities compared to methanogenic communities in all five studied locations. [ABSTRACT FROM AUTHOR]

Abstract: In October 2008, we measured temperature and salinity in hot, hypersaline brine filling the Atlantis II and Discovery Deeps on the Red Sea spreading center west of Jeddah, Saudi Arabia. In agreement with previous observations in the Atlantis II Deep, we found a stack of four convective layers with vertically uniform temperature profiles separated by thin interfaces with high vertical temperature gradients. Temperature in the thick lower convective layer in the Atlantis II Deep continued to slowly increase at 0.1°C/year since the last observations in 1997. Previously published data show that the temperature of all four convective layers increased since the 1960s at the same rate, from which we infer that diffusive vertical heat flux between convective layers is rapid on time scales of 3–5 years and, thus, heat is lost from the brine pools to overlying Red Sea Deep Water. Heat budgets suggest that the heat flux from hydrothermal venting has decreased from 0.54GW to 0.18GW since 1966. A tow-yo survey found that temperature in the upper convective layers changes about 0.2°C over 5–6km but the temperature in the lower brine layer remains constant. Temperature in the lower convective layer in the Discovery Deep remains unchanged at 48°C. To explain these results, we hypothesize that heat flux from a hydrothermal vent in the floor of the Discovery Deep has been stable for 40 years, whereas temperature of the brine in the Atlantis II Deep is adjusting to the change in hydrothermal heat flux from the vent in the Southwest Basin. We found no changes in the upper transition layer at 1900–1990m depth that appeared between 1976 and 1992 and suggest that this layer originated from the seafloor elsewhere in the rift. [Copyright &y& Elsevier]

Abstract: The Monarch, a mountain in southeastern British Columbia, exposes the Cathedral Escarpment, which abuts the basinal Burgess Shale and an underlying limestone, the upper Takakkaw Tongue. At one section, a dolomitized megabreccia, representing an intraformational collapsed cavern, forms the Cathedral Escarpment wall. The megabreccia is cross-cut by the subsequent megatruncation event marking the collapse of the Cathedral margin, which supports previous hypotheses that dolomitizing brines were active at the Cathedral margin, prior to and during deposition of the Burgess Shale. Epiphyton-rich carbonate megablocks within the basinal strata, previously interpreted as olistoliths spalled off the Escarpment, are unlike the peritidal strata comprising the Cathedral margin and are more likely deep-water carbonate mud mounds that grew in the basin near the Escarpment face. Evidence for early brines in basinal environments at this locality includes lenses of dark, magnesium-rich clinochlore (>30 wt.% MgO vs ~2% in background shales) within the Burgess Shale at the Escarpment wall. These clinochlores are interpreted as precipitates from seafloor brine seeps analogous to ones currently found at the foot of the Florida Escarpment and on the distal shelf and slope of the northern Gulf of Mexico. Like the Gulf examples, the brines formed standing pools on the Cambrian seafloor and hosted abundant organisms in a narrow fringe around their peripheries but not within the pools. Sponges, priapulid worms, hyoliths, linguliform brachiopods and trilobites predominate. Shales only a few metres basinward from the pools are nearly barren of animal fossils. Increased biologic productivity localized at the seeps would account for the patchy distribution of fossils in the Burgess Shale in a narrow zone close to the Escarpment. The discovery of brine seep deposits and associated fringing communities at The Monarch shows that at least some assemblages in the Burgess Shale were preserved within their habitat and were supported by chemosynthesis. [Copyright &y& Elsevier]

Eight Zn–Pb–Cu massive sulphide deposits that appear to have formed on the sea floor (seven in Spain, one in Tasmania) are believed to have been precipitated in brine pools, based on the salinities and temperatures of fluid inclusions in underlying stockworks. Comparing the geological features of these deposits with those of the Zn–Pb–Cu massive sulphide ores of the Hokuroku Basin, Japan, which have formed as mounds from buoyant fluids of low salinity, shows that brine pool deposits have: (1) potentially very large size and tonnage, and high aspect ratio, (2) higher Zn/Cu and Fe/Cu values, (3) no evidence of chimneys, (4) relatively abundant framboidal pyrite and primary mineral banding, (5) reduced mineral assemblages (pyrite-arsenopyrite/pyrrhotite), and minor or rare barite in the massive sulphide, (6) associated stratiform and/or vein carbonates, (7) relatively unimportant zone refining, (8) lack of vertical variation in sphalerite and sulphur isotopic compositions, and (9) evidence of local bacterial sulphate reduction. Application of these criteria to the Rosebery deposit in Tasmania, for which there are no fluid inclusion data, leads to the conclusion that the southern section was deposited as separate lenses in a brine-filled basin or basins. Other potential candidates include Brunswick no. 12 and Heath Steele (Canada), Woodlawn and Captains Flat (New South Wales), Hercules and Que River (Tasmania), and Tharsis and the orebodies at Aljustrel (Spain and Portugal). Recently published fluid inclusion data for Gacun (China) and Mount Chalmers (Queensland) suggest that not all ores deposited from highly saline fluids have reduced mineral assemblages. [Copyright &y& Elsevier]

Hypersaline environments are found around the world, above and below ground, and many are exposed to hydrocarbons on a continuous or a frequent basis. Some surface hypersaline environments are exposed to hydrocarbons because they have active petroleum seeps while others are exposed because of oil exploration and production, or nearby human activities. Many oil reservoirs overlie highly saline connate water, and some national oil reserves are stored in salt caverns. Surface hypersaline ecosystems contain consortia of halophilic and halotolerant microorganisms that decompose organic compounds including hydrocarbons, and subterranean ones are likely to contain the same. However, the rates and extents of hydrocarbon biodegradation are poorly understood in such ecosystems. Here we describe hypersaline environments potentially or likely to become contaminated with hydrocarbons, including perennial and transient environments above and below ground, and discuss what is known about the microbes degrading hydrocarbons and the extent of their activities. We also discuss what limits the microbial hydrocarbon degradation in hypersaline environments and whether there are opportunities for inhibiting (oil storage) or stimulating (oil spills) such biodegradation as the situation requires. Unlabelled Image • Many environments are permanently or transiently hypersaline. • Hypersaline ecosystems can be exposed to hydrocarbons. • Biodegradation is slow in hypersaline ecosystems, and it is not clear why. • It would be beneficial to inhibit biodegradation in reservoirs and storage caverns. • As would stimulating biodegradation of oil spills in surface hypersaline systems [ABSTRACT FROM AUTHOR]

Background: Microbial species in the brine pools of the Red Sea and the brine pool-seawater interfaces are exposed to high temperature, high salinity, low oxygen levels and high concentrations of heavy metals. As adaptations to these harsh conditions require a large suite of secondary metabolites, these microbes have a huge potential as a source of novel anticancer molecules. Methods: A total of 60 ethyl-acetate extracts of newly isolated strains from extreme environments of the Red-Sea were isolated and tested against several human cancer cell lines for potential cytotoxic and apoptotic activities. Results: Isolates from the Erba brine-pool accounted for 50% of active bacterial extracts capable of inducing 30% or greater inhibition of cell growth. Among the 60 extracts screened, seven showed selectivity towards triple negative BT20 cells compared to normal fibroblasts. Conclusion: In this study, we identified several extracts able to induce caspase-dependent apoptosis in various cancer cell lines. Further investigations and isolation of the active compounds of these Red Sea brine pool microbes may offer a chemotherapeutic potential for cancers with limited treatment options. [ABSTRACT FROM AUTHOR]

Two moderately halophilic marine bacterial strains of the family Rhodobacteraceae, designated ZGT108 T and ZGT118 T , were isolated from the brine-seawater interface at Erba Deep in the Red Sea (Saudi Arabia). Cells of both strains were aerobic, rod-shaped, non-motile, and Gram-stain-negative. The sequence similarity of the 16S rRNA genes of strains ZGT108 T and ZGT118 T was 94.9 %. The highest 16S rRNA gene sequence similarity of strain ZGT108 T to its closest relative, Ruegeria conchae JCM 17315 T , was 98.9 %, while the 16S rRNA gene of ZGT118 T was most closely related to that of Ruegeria intermedia LMG 25539 T (97.7 % similarity). The sizes of the draft genomes as presented here are 4 258 055 bp (strain ZGT108 T ) and 4 012 109 bp (strain ZGT118 T ), and the G+C contents of the draft genomes are 56.68 mol% (ZGT108 T ) and 62.94 mol% (ZGT108 T ). The combined physiological, biochemical, phylogenetic and genotypic data supported placement of both strains in the genus Ruegeria and indicated that the two strains are distinct from each other as well as from all other members in the genus Ruegeria. This was also confirmed by low DNA-DNA hybridization values (<43.6 %) and low ANI values (<91.8 %) between both strains and the most closely related Ruegeria species. Therefore, we propose two novel species in the genus Ruegeria to accommodate these novel isolates: Ruegeriaprofundi sp. nov. (type strain ZGT108 T =JCM 19518 T =ACCC 19861 T ) and Ruegeriamarisrubri sp. nov. (type strain ZGT118 T =JCM 19519 T =ACCC 19862 T ).