Hadal Amphipods Research Articles

Development of Single-Nucleotide Polymorphism Markers and Population Genetic Analysis of the Hadal Amphipod Alicella gigantea across the Mariana and New Britain Trenches

Alicella gigantea, the largest amphipod scavengers found to date, play key roles in the food web of the hadal ecosystem. However, the genetic structure of A. gigantea populations among different trenches has not been reported yet. In this study, SNP (single-nucleotide polymorphism) markers were developed for three A. gigantea geographic populations collected from the southern Mariana Trench (SMT), the central New Britain Trench (CNBT), and the eastern New Britain Trench (ENBT), based on the SLAF-seq (specific locus amplified fragment sequencing) technology. A total of 570,168 filtered SNPs were screened out for subsequent population genetic analysis. Results showed that the inbreeding levels across the three geographic populations were relatively low, and the genomic inbreeding coefficients of the three populations were similar in magnitude. Based on the results of phylogenetic analysis, population structure analysis, and principal component analysis, it is believed that the three A. gigantea geographic populations belong to the same population, and the kinship relationship between the ENBT and CNBT populations is close. Moreover, the differential candidate adaptive sites on the SNPs suggest that there may be variations in metabolic rates among the three geographic populations, possibly linked to differences in food availability and sources in different trenches, ultimately resulting in different survival strategies in A. gigantea populations within distinct trenches. Compared with the Mariana Trench, the New Britain Trench has a richer organic matter input, and it is speculated that the A. gigantea Mariana Trench population may adopt a lower metabolic rate to cope with the harsher environment of nutrient deficiency.

Depth and predation regulate consumption of dolphin carcasses in the hadal zone

Natural whale falls and falls of smaller-sized food have been reported for more than 30 years and are known to be important sources of nutrients and organic matter for the seafloor community. However, the associated species composition and ecological processes during such events in the hadal zone were unknown. Therefore, we aimed to determine the impact of depth and predation on the hadal community during the early stages of a food-fall event. This is the first study to investigate the preliminary stages of the two deepest artificial dolphin-fall events in the Philippine Basin (PB) (∼7729 m) and Mariana Trench (MT) (∼8200 m). A total of nine dives were conducted over a period of 86 days (PB) and 50 days (MT) using the deep-sea manned submersible “Fendouzhe.” Our observations in the PB indicated that the first stage (mobile-scavenger) was controlled by the feeding ecology of hadal amphipods and snailfish. In contrast, the absence of predatory snailfish in the MT enhanced the degradation rate of the carcass compared to that in the PB. Most soft tissues were entirely consumed by scavenging amphipods within days of the event in the MT, whereas in the PB, amphipods were observed taking a feeding hiatus on Day 10 to escape snailfish predation. The second stage (enrichment-opportunist), which hosted few grazing faunas, began at different times in each location and overlapped with the mobile-scavenger stage. Dolphin carcasses, being smaller than those of full-grown whales, can only sustain a large community of scavenging amphipods, and indirectly, predatory snailfish. After the first stage, the dispersed organic matter and limited lipid content in dolphin bones were likely insufficient to sustain an active grazing community or the chemosynthetic community that typically follows. We concluded that water depth influences the successional stages and decomposition rate of food falls in the hadal zone by controlling the dynamic relationship between prey and predators. Our study elucidates the ecology of food-fall events in the hadal zone and highlights the key differences in food-fall events at different depths.

Comparably Characterizing the Gut Microbial Communities of Amphipods from Littoral to Hadal Zones

Amphipods are an important group of invertebrates in marine ecosystems due to their high abundance and diversity. As an essential part of the marine food web, amphipods play a vital role in nutrient recycling and provide large amounts of detritus-derived fine-particulate organic matter for other invertebrates. Although the importance of gut microbiota and the necessity to consider them has been increasingly recognized, the gut microbial community and diversity of amphipods have not been well studied. Here, we comparatively studied the gut microbiota of diverse amphipod species inhabiting from coastal to hadopelagic zones. The results showed that four phyla, including Proteobacteria, Firmicutes, Bacteroidota, and Actinobacteriota, occupied more than 90% of the total microbes in the studied amphipod guts, with Firmicutes being dominant in the hadal amphipods. The gut microbiome of amphipods from the hadal zone displayed the lowest richness, lowest diversity, and shared few microorganisms with the surrounding seawater compared to others. Amphipods in different inhabiting regions have discriminant taxa for their gut microbial communities. Taken together, amphipod gut microbiota was affected by both biological and abiotic factors, yet these factors are not independent. This article provides us with a further understanding of the structure and characteristics of the gut microbiota of invertebrate organisms.

Metal Accumulations in Two Extreme-Environment Amphipods, Hadal Eurythenes gryllus and Antarctic Pseudorchomene plebs

The hadal zone and Antarctic Ocean are two of the least-explored habitats. Knowledge about human impacts on these two extreme environments is limited. Here, we analyzed the metal accumulations of two amphipod species, Eurythenes gryllus, from the Mariana Trench (6040 m, 11.36° N, 142.41° E) in the West Pacific Ocean, and Pseudorchomene plebs, from the Ross Sea (600 m, 77.12° S, 167,67° E) in the Antarctic. Bioaccumulation of thirteen elements (Na, Mg, K, Ca, Cu, Fe, Al, Cr, Mn, Zn, As, Se, and Cd) in three tissues (exoskeleton, leg muscle, and gut) of the two amphipods was investigated using inductively coupled plasma mass spectrometry (ICP-MS). Comparing the trace element concentrations between the different amphipoda species, we found higher element concentrations in the Antarctic amphipod, and an oligotrophication of the Mariana Trench. The concentrations of Cu, Zn, As, and Se in the three tissues all had a significant difference in abundance, and the Cd in the gut of P. plebs was comparably higher than that in E. gryllus, consistent with special environment adaptation. Compared with non-abyssal and shallow water decapoda and amphipoda species, hadal amphipods possessed comparably higher concentrations of Cd and Cr elements and displayed a very high environmental specificity for amphipods’ metal-element bioaccumulation strategy. This study reveals the amphipods of remote and uncontaminated areas as potential indicator species for metal-element bioaccumulation to measure anthropogenic impacts.

Occurrence of polybrominated diphenyl ethers and benzotriazole UV stabilizers in the hadal amphipod Hirondellea gigas

The accumulation of polybrominated diphenyl ethers (PBDEs) and benzotriazole UV stabilizers (BZT-UVs) were examined in the hadal amphipod Hirondellea gigas caught from a near-land trench off the Japan island (9200 m). H.gigas were collected from two distinct sites: one is located at the outlet of submarinecanyons directly connected to land and the other is apart from the outlet and geographically isolated from the first site. The level of the PBDEs in H.gigas at the canyon outlet (mean 219ng/g lipid weight (l.w.)) was significantly higher than that in the isolated site (mean 42ng/g l.w.) and BZT-UVs were only detected within the first site (mean 1.5 ng/g wet weight). In addition to vertical transport from the surface water, near-land trenches associated with submarine canyons and troughs may have more complex influx of contaminants through horizontal transportation from the land, resulting in more severe contamination.

Morphological and molecular evolution of hadal amphipod's eggs provides insights into embryogenesis under high hydrostatic pressure.

Hadal zones are unique habitats characterized by high hydrostatic pressure (HHP) and scarce food supplies. The ability of eggs of species dwelling in hadal zones to develop into normal embryo under high hydrostatic pressure is an important evolutionary and developmental trait. However, the mechanisms underlying the development of eggs of hadal-dwelling species remain unknown due to the difficulty of sampling ovigerous females. Here, morphological and transcriptome analyses of eggs of the “supergiant” amphipod Alicella gigantea collected from the New Britain Trench were conducted. The morphology of A. gigantea eggs, including size, was assessed and the ultrastructure of the eggshell was investigated by scanning electron microscopy. Transcriptome sequencing and molecular adaptive evolution analysis of A. gigantea eggs showed that, as compared with shallow-water Gammarus species, genes exhibiting accelerated evolution and the positively selected genes were mostly related to pathways associated with “mitosis” and “chitin-based embryonic cuticle biosynthetic process”, suggesting that “normal mitosis maintenance” and “cuticle development and protection” are the two main adaptation strategies for survival of eggs in hadal environments. In addition, the concentration of trimethylamine oxide (TMAO), an important osmotic regulator, was significantly higher in the eggs of hadal amphipods as compared to those of shallow-water species, which might promote the eggs’ adaptation abilities. Morphological identification, evolutionary analysis, and the trimethylamine oxide concentration of A. gigantea eggs will facilitate a comprehensive overview of the piezophilic adaptation of embryos in hadal environments and provide a strategy to analyze embryogenesis under high hydrostatic pressure.

Unusual bromine enrichment in the gastric mill and setae of the hadal amphipod Hirondellea gigas.

The hadal amphipod Hirondellea gigas is an emblematic animal of the Pacific trenches, and has a number of special adaptations to thrive in this 'extreme' environment, which includes the deepest part of the Earth's ocean. One such adaptation that has been suggested is the presence of an 'aluminum gel shield' on the surface of its body in order to prevent the dissolution of calcitic exoskeleton below the carbonate compensation depth. However, this has not been investigated under experimental conditions that sufficiently prevent aluminum artefacts, and the possibility of other elements with similar characteristic X-ray energy as aluminum (such as bromine) has not been considered. Here, we show with new electron microscopy data gathered under optimized conditions to minimize aluminum artefacts that H. gigas actually does not have an aluminum shield-instead many parts of its body are enriched in bromine, particularly gastric ossicles and setae. Results from elemental analyses pointed to the use of calcite partially substituted with magnesium by H. gigas in its exoskeleton, in order to suppress dissolution. Our results exemplify the necessity of careful sample preparation and analysis of the signals in energy-dispersive X-ray spectroscopic analysis, and the importance of analyses at different electron energies.

Trimethylamine N-Oxide (TMAO) and Trimethylamine (TMA) Determinations of Two Hadal Amphipods

Hadal trenches are a unique habitat with high hydrostatic pressure, low temperature and scarce food supplies. Amphipods are the dominant scavenging metazoan species in this ecosystem. Trimethylamine (TMA) and trimethylamine oxide (TMAO) have been shown to play important roles in regulating osmotic pressure in mammals, hadal dwellers and even microbes. However, the distributions of TMAO and TMA concentrations of hadal animals among different tissues have not been reported so far. Here, the TMAO and TMA contents of eight tissues of two hadal amphipods, Hirondellea gigas and Alicella gigantea from the Mariana Trench and the New Britain Trench, were detected by using the ultrahigh performance liquid chromatography–mass spectrometry (UPLC-MS/MS) method. Compared with the shallow water Decapoda, Penaeus vannamei, the hadal amphipods possessed significantly higher TMAO concentrations and a similar level of TMA in all the detected tissues. A higher level of TMAO was detected in the external organs (such as the eye and exoskeleton) for both of the two hadal amphipods, which indicated that the TMAO concentration was not evenly distributed, although the same hydrostatic pressure existed in the outer and internal organs. Moreover, a strong positive correlation was found between the concentrations of TMAO and TMA in the two hadal amphipods. In addition, evolutionary analysis regarding FMO3, the enzyme to convert TMA into TMAO, was also conducted. Three positive selected sites in the conserved region and two specific mutation sites in two conserved motifs were found in the A. gigantea FMO3 gene. Combined together, this study supports the important role of TMAO for the environmental adaptability of hadal amphipods and speculates on the molecular evolution and protein structure of FMO3 in hadal species.

The Adaptive Evolution and Gigantism Mechanisms of the Hadal “Supergiant” Amphipod Alicella gigantea

Hadal trenches are commonly referred to as the deepest areas in the ocean and are characterized by extreme environmental conditions such as high hydrostatic pressures and very limited food supplies. Amphipods are considered the dominant scavengers in the hadal food web. Alicella gigantea is the largest hadal amphipod and, as such, has attracted a lot of attention. However, the adaptive evolution and gigantism mechanisms of the hadal “supergiant” remain unknown. In this study, the whole-body transcriptome analysis was conducted regarding the two hadal amphipods, one being the largest sized species A. gigantea from the New Britain Trench and another the small-sized species Bathycallisoma schellenbergi from the Marceau Trench. The size and weight measurement of the two hadal amphipods revealed that the growth of A. gigantea was comparatively much faster than that of B. schellenbergi. Phylogenetic analyses showed that A. gigantea and B. schellenbergi were clustered into a Lysianassoidea clade, and were distinct from the Gammaroidea consisting of shallow-water Gammarus species. Codon substitution analyses revealed that “response to starvation,” “glycerolipid metabolism,” and “meiosis” pathways were enriched among the positively selected genes (PSGs) of the two hadal amphipods, suggesting that hadal amphipods are subjected to intense food shortage and the pathways are the main adaptation strategies to survive in the hadal environment. To elucidate the mechanisms underlying the gigantism of A. gigantea, small-sized amphipods were used as the background for evolutionary analysis, we found the seven PSGs that were ultimately related to growth and proliferation. In addition, the evolutionary rate of the gene ontology (GO) term “growth regulation” was significantly higher in A. gigantea than in small-sized amphipods. By combining, those points might be the possible gigantism mechanisms of the hadal “supergiant” A. gigantea.

Gut Microbial Divergence Between Three Hadal Amphipod Species from the Isolated Hadal Trenches.

Amphipods are the dominant scavenging metazoan species in the hadal trenches at water depths below 6,000 m. The gut microbiota have been considered to be contribution to the adaptation of deep-sea organisms; however, few comparative analyses of animal gut microbiota between different isolated hadal environments have been done so far. Here, we employed high-throughput 16S rRNA sequencing to compare the gut microbial taxonomic composition and functional potential diversity of three hadal amphipod species, Hirondellea gigas, Bathycallisoma schellenbergi, and Alicella gigantea, collected from the Mariana Trench, Marceau Trench, and New Britain Trench in the Pacific Ocean, respectively. Results showed that different community compositions were detected across all the amphipod specimens based on the analyses of alpha-diversity, hierarchical cluster tree, and PCoA (principal coordinate analysis). Moreover, almost no correlation was observed between genera overrepresented in different amphipods by microbe-microbe correlations analysis, which suggested that the colonization of symbionts were host-specific. At genus level, Psychromonas was dominant in H. gigas, and Candidatus Hepatoplasma was overall dominant in A. gigantea and B. schellenbergi. Comparison of the functional potential showed that, though three hadal amphipod species shared the same predominant functional pathways, the abundances of those most shared pathways showed distinct differences across all the specimens. These findings pointed to the enrichment of particular functional pathways in the gut microbiota of the different isolated trench amphipods. Moreover, in terms of species relative abundance, alpha-diversity and beta-diversity, there was high similarity of gut microbiota between the two A. gigantea populations, which dwelled in two different localities of the same hadal trench. Altogether, this study provides an initial investigation into the gut-microbial interactions and evolution at the hadal depths within amphipod. Each of these three amphipod species would be a model taxa for future studies investigating the influence habitat difference and geography on gut-microbial communities.

Metagenomic Insights Into the Structure and Function of Intestinal Microbiota of the Hadal Amphipods.

Hadal trenches are the deepest known areas of the ocean. Amphipods are considered to be the dominant scavengers in the hadal food webs. The studies on the structure and function of the hadal intestinal microbiotas are largely lacking. Here, the intestinal microbiotas of three hadal amphipods, Hirondellea gigas, Scopelocheirus schellenbergi, and Alicella gigantea, from Mariana Trench, Marceau Trench, and New Britain Trench, respectively, were investigated. The taxonomic analysis identified 358 microbial genera commonly shared within the three amphipods. Different amphipod species possessed their own characteristic dominant microbial component, Psychromonas in H. gigas and Candidatus Hepatoplasma in A. gigantea and S. schellenbergi. Functional composition analysis showed that “Carbohydrate Metabolism,” “Lipid Metabolism,” “Cell Motility,” “Replication and Repair,” and “Membrane Transport” were among the most represented Gene Ontology (GO) Categories in the gut microbiotas. To test the possible functions of “Bacterial Chemotaxis” within the “Cell Motility” category, the methyl-accepting chemotaxis protein (MCP) gene involved in the “Bacterial Chemotaxis” pathway was obtained and used for swarming motility assays. Results showed that bacteria transformed with the gut bacterial MCP gene showed significantly faster growths compared with the control group, suggesting MCP promoted the bacterial swimming capability and nutrient utilization ability. This result suggested that hadal gut microbes could promote their survival in poor nutrient conditions by enhancing chemotaxis and motility. In addition, large quantities of probiotic genera were detected in the hadal amphipod gut microbiotas, which indicated that those probiotics would be possible contributors for promoting the host’s growth and development, which could facilitate adaptation of hadal amphipods to the extreme environment.

Trace Elemental Analysis of the Exoskeleton, Leg Muscle, and Gut of Three Hadal Amphipods.

Hadal trenches are the deepest areas worldwide. Amphipods are considered a key factor in hadal ecosystems because of their important impacts on the hadal environment. Amphipods have benthic habits, and therefore, serve as good metal biomonitors. However, little is known about the hadal amphipod metal accumulations. In the present study, Alicella gigantea, Hirondellea gigas, and Scopelocheirus schellenbergi were sampled from the New Britain Trench (8824m, 7.02S 149.16E), Mariana Trench (10,839m, 11.38N 142.42E), and Marceau Trench (6690m, 1.42N 148.74E) in the West Pacific Ocean, respectively. The elemental concentrations of the three hadal amphipods were subsequently investigated. Nine trace elements (V, Cr, Mn, Co, Ni, Se, Mo, Ag, and Cd) of three tissues (exoskeleton, leg muscle, and gut) of the hadal amphipods were detected by using inductively coupled plasma mass spectrometry (ICP-MS) method. The concentrations of Cr, Cd, and Mn were comparably higher among those nine examined elements. The greatest accumulations of the elements Cr, Ag, and V in the exoskeleton and leg muscle were observed in H. gigas, and elements Mn, Co, and Se showed the highest accumulations in the gut in H. gigas among the three hadal amphipods. In addition, comparisons of the leg muscle trace element accumulation between the hadal amphipods and non-abyssal and shallow water decapoda and amphipoda species showed that the hadal amphipods possessed comparably higher concentrations of the trace elements Cd, Co, Mo, Ag, and V. This finding suggested a bottom-up effect of food availability and indicated the effects of human activities within the hadal environments. This study reveals the trace element bio-accumulation of three hadal amphipods, and suggests that deep-sea amphipods are potential indicator species for trace element bioavailability in the deep-sea environment.

Scavenging amphipods from the Wallaby-Zenith Fracture Zone: Extending the hadal paradigm beyond subduction trenches

Our understanding of the ecology of the hadal zone (> 6000 m depth) is based solely on subduction trenches, leaving other geomorphological features, such as fracture zones, troughs, and basins, understudied. To address this knowledge gap, the Wallaby-Zenith Fracture Zone, Indian Ocean (WZFZ; ~ 22°S, 102°E; maximum depth 6625 m measured during Expedition SO258) was studied using free-fall baited landers. We assessed the amphipod distribution and community assemblage of this non-subduction hadal feature and compared it to subduction hadal features. Eleven species were identified across the abyssal-hadal transition zone using a paired morphological and DNA barcoding approach. The community composition was found to change gradually from abyssal to hadal depths, which contrasts with the ecotone shift characteristic of subduction trenches. A large population of Bathycallisoma schellenbergi (Birstein & Vinogradov, 1958), a quintessential hadal amphipod, was present at the flat bottom of the WZFZ. Further, an mtDNA phylogeny resolved a degree of phylogeographic structure between the B. schellenbergi WZFZ population and four previously sampled Pacific Ocean subduction trench populations, indicating these features are not interconnected through ongoing gene flow. Combined, these data indicate that some amphipods have far broader distributions than previously understood, with some species present in both hadal subduction trenches and non-subduction fracture zones and basins interspersed across the abyssal plains. This initial exploration highlights that whilst non-subduction features are an overlooked minor fraction of the total hadal area, they are essential to our understanding of the ecological and evolutionary dynamics across the hadal zone.

Ontogenetic diet change of hadal amphipods in the New Britain Trench revealed by fatty acid biomarker and stable isotope ratio

The hadal zone (from 6 to 11 km depth) is one of the least explored habitats on Earth, and the knowledge about trophic ecology of hadal organisms is limited. Here, we analyzed fatty acid biomarker and stable isotope ratios of the amphipod species (Alicella gigantean) collected in the New Britain Trench (8225 m depth) in the southwestern Pacific Ocean. A total of 50 samples were divided into five developmental stages according to body length (1–129 cm): Stages I (juvenile), II (sub-adult) and III/IV/V (adult). Stages III, IV and V were further divided according to sex. Mean total fatty acid content for stage I (237.7 ± 105.6 mg g-1) was significantly lower than that for stages II to V (476.4 ± 49.0 mg g-1). The δ15N (14.7–18.4‰) positively correlated with the body length, whereas the EPA/DHA (C20:5ω3/C22:6ω3) ratio (0.09–0.51) negatively correlated with the body length. These results suggested an increase of trophic level with growth. Principal component analysis (PCA) separated major fatty acids into three clusters: (1) C18:1ω9, C16:1ω7, C14:1ω5 (indicative of carrion-based diet); (2) C14:0, C15:0, isoC15:0, C16:0, C17:0, isoC17:0, anteisoC17:0, C18:1ω7, C18:0, C20:1ω9, and C22:1ω9 (bacteria/zooplankton-based diet); and (3) C20:4ω6, C20:5ω3, C22:6ω3 and C22:5ω6 (benthic fauna/algae-based diet). The PCA result revealed that stage I-individuals consumed more bacteria/zooplankton debris, stages II to IV-individuals primarily preyed on carrion, while stage V-individuals had more diverse diet (benthic fauna/algae and carrion). Female amphipods had higher proportions of monounsaturated fatty acids, lower proportions of polyunsaturated fatty acids, and higher δ15N compared to male amphipods, suggesting that female individuals rely more on carrion from higher trophic organisms. This sex-related difference, however, diminished from stage III to V. Overall, our fatty acid and stable isotope data suggest an ontogenetic diet shift for hadal amphipods that may contribute to their success in the deepest ocean.

High species richness of Northwest Pacific deep-sea amphipods revealed through DNA barcoding

Although deep sea has become a subject of intense studies in the recent years, our knowledge of the diversity of benthic organisms, especially at the molecular level, is far from complete. In the present study we focused on Amphipoda, known to be an abundant and diverse component of the marine zoobenthos. COI barcoding of an extensive collection of abyssal and hadal amphipods from the Kuril-Kamchatka Trench (KKT) area revealed 133 Molecular Operational Taxonomical Units (MOTUs). The richness of species varied between stations, with the highest observed in the areas where high organic carbon (Corg) content was recorded. Among the recognized MOTUs as many as 42% were singletons and only 28 were found at three or more stations. Their distribution was variable with seven taxa occurring at the most distant stations situated across the KKT. This shows that the trench does not constitute a barrier for the distribution of certain species. However, it may limit the gene flow in the case of others. The most common MOTUs were divided into three bathymetric categories: typically abyssal and hadal species as well as eurybathic ones inhabiting both the abyssal and hadal depth zones.

The complete mitochondrial genome of the largest amphipod, Alicella gigantea: Insight into its phylogenetic relationships and deep sea adaptive characters

Alicella gigantea (Alicelloidae) is a scavenger with the largest body size among amphipods. It is a participant in the foodweb of deepsea ecosystem and distributed with vast bathymetric and geographic ranges. In this study, the mitochondrial genome of A. gigantea was completely assembled and characterized. The complete sequence has a total length of 16,851 bp, comprising the usual eukaryotic components, with 13 protein-coding genes (PCGs), 2 ribosomal RNA genes (rRNAs), 22 transfer RNA genes (tRNAs), and 2 noncoding control regions (CRs). The gene rearrangement and reverse nucleotide strand bias of its mitochondrial genome are similar to those observed in the deepsea amphipod Eurythenes maldoror (Eurytheneidae), but different from the characters of Halice sp. MT-2017 (Dexaminoidea), an inhabitant of a deeper environment. Phylogenetic analysis indicates that A. gigantea occupies the basal branch of deepsea species—E. maldoror and Hirondellea gigas. This phylogeny supports the hypothesis that the evolution of hadal amphipods has undergone a transition from the abyssal depth. Compared to 41 available shallow water equivalents, the four accessible mitochondrial genomes from the deep sea, including the one produced in this study, show significantly fewer charged amino acids in the 13 PCGs, which suggests an adaption to the deepsea environment.

Comparative analysis of the gut microbial communities between two dominant amphipods from the Challenger Deep, Mariana Trench

The gut microbiota is contributable to the adaptation and health of deep-sea organisms and has been revealed to be affected by many factors, especially the host species and diet. Here, we employed high-throughput sequencing of 16S rDNA to compare the gut microbial composition and diversity of two hadal amphipod species, Hirondellea gigas and Halice sp. MT-2017, which are predominant inhabitants in the Challenger Deep. Results showed that Tenericutes and Proteobacteria were the most abundant phyla, occupying more than 50% of total phyla, across all the specimens. At genus level, “Candidatus Hepatoplasma” was overall dominant, followed by Psychromonas in H. gigas and Psychrobacter in Halice sp. MT-2017 respectively. Notably, though two hadal amphipod species shared the same predominant symbiont of genus “Ca. Hepatoplasma”, they were classified into two different OTUs, which suggested that the colonization of symbionts were host-specific. Moreover, in terms of species relative abundance, alpha-diversity and beta-diversity, there was still a significant divergence of gut microbiota between two hadal amphipod species, which dwelled in the same extreme environment. Altogether, the results dropped a hint to the importance of symbiont genus of “Ca. Hepatoplasma” to amphipod survival in the hadal zone.

Penetration of Bomb 14C Into the Deepest Ocean Trench

AbstractThe food source of hadal endemic fauna provides an insight into the carbon cycle in trenches and a biological adaptation to the impoverished and harsh trench environment. Here, we present the first Δ14C results of hadal amphipods from three trenches in the Pacific to define the organic matter source in these remote ecosystems. Amphipod muscle tissues contain a bomb 14C signature (Δ14C from 10 ± 2‰ to 65 ± 2‰), thereby revealing a dietary preference for labile and fresh organic matter derived from the surface water. Thus, the carbon cycle in the deepest ocean trench has a tight linkage with the surface ocean via the food chain. The bomb 14C dating result suggests that hadal amphipods have a low tissue turnover rate and an unexpectedly long lifetime (>10 years), at more than 4 times higher than the common longevity (~2 years) of amphipods in shallow waters.

Characterization of the mitochondrial genome of an ancient amphipod Halice sp. MT-2017 (Pardaliscidae) from 10,908\u2009m in the Mariana Trench

Small amphipods (Halice sp. MT-2017) with body length <1 cm were collected from the Challenger Deep (~10,920 m below sea level). The divergence time of their lineage was approximately 109 Mya, making this group ancient compared to others under study. The mitochondrial genome of Halice sp. shared the usual gene components of metazoans, comprising 13 protein coding genes (PCGs), 22 transfer RNAs (tRNAs), and 2 ribosomal RNAs (rRNAs). The arrangement of these genes, however, differed greatly from that of other amphipods. Of the 15 genes that were rearranged with respect to the pancrustacean gene pattern, 12 genes (2 PCGs, 2 rRNAs, and 8 tRNAs) were both translocated and strand-reversed. In contrast, the mitochondrial genomes in other amphipods never show so many reordered genes, and in most instances, only tRNAs were involved in strand-reversion-coupled translocation. Other characteristics, including reversed strand nucleotide composition bias, relatively higher composition of non-polar amino acids, and lower evolutionary rate, were also identified. Interestingly, the latter two features were shared with another hadal amphipod, Hirondellea gigas, suggesting their possible associations with the adaptation to deep-sea extreme habitats. Overall, our data provided a useful resource for future studies on the evolutionary and adaptive mechanisms of hadal faunas.

Gut Microbial Divergence between Two Populations of the Hadal Amphipod Hirondellea gigas.

Hadal environments sustain diverse microorganisms. A few studies have investigated hadal microbial communities consisting of free-living or particle-associated bacteria and archaea. However, animal-associated microbial communities in hadal environments remain largely unexplored, and comparative analyses of animal gut microbiota between two isolated hadal environments have never been done so far. In the present study, 228 Gb of gut metagenomes of the giant amphipod Hirondellea gigas from two hadal trenches, the Mariana Trench and Japan Trench, were sequenced and analyzed. Taxonomic analysis identified 49 microbial genera commonly shared by the gut microbiota of the two H. gigas populations. However, the results of statistical analysis, in congruency with the alpha and beta diversity analyses, revealed significant differences in gut microbial composition across the two trenches. Abundance variation of Psychromonas, Propionibacterium, and Pseudoalteromonas species was observed. Microbial cooccurrence was demonstrated for microbes that were overrepresented in the Mariana trench. Comparison of functional potential showed that the percentage of carbohydrate metabolic genes among the total microbial genes was significantly higher in the guts of H. gigas specimens from the Mariana Trench. Integrating carbon input information and geological characters of the two hadal trenches, we propose that the differences in the community structure might be due to several selective factors, such as environmental variations and microbial interactions.IMPORTANCE The taxonomic composition and functional potential of animal gut microbiota in deep-sea environments remain largely unknown. Here, by performing comparative metagenomics, we suggest that the gut microbial compositions of two Hirondellea gigas populations from the Mariana Trench and the Japan Trench have undergone significant divergence. Through analyses of functional potentials and microbe-microbe correlations, our findings shed light on the contributions of animal gut microbiota to host adaptation to hadal environments.