Host individual and gut location are more important in gut microbiota community composition than temporal variation in the marine herbivorous fish Kyphosus sydneyanus.

BACKGROUND: Gut microbiota play a key role in the nutrition of many marine herbivorous fishes through hindgut fermentation of seaweed. Gut microbiota composition in the herbivorous fish Kyphosus sydneyanus (family Kyphosidae) varies between individuals and gut sections, raising two questions: (i) is community composition stable over time, especially given seasonal shifts in storage metabolites of dietary brown algae, and (ii) what processes influence community assembly in the hindgut? RESULTS: We examined variation in community composition in gut lumen and mucosa samples from three hindgut sections of K. sydneyanus collected at various time points in 2020 and 2021 from reefs near Great Barrier Island, New Zealand. 16S rRNA gene analysis was used to characterize microbial community composition, diversity and estimated density. Differences in community composition between gut sections remained relatively stable over time, with little evidence of temporal variation. Clostridia dominated the proximal hindgut sections and Bacteroidia the most distal section. Differences were detected in microbial composition between lumen and mucosa, especially at genus level. CONCLUSIONS: High variation in community composition and estimated bacterial density among individual fish combined with low variation in community composition temporally suggests that initial community assembly involved environmental selection and random sampling/neutral effects. Community stability following colonisation could also be influenced by historical contingency, where early colonizing members of the community may have a selective advantage. The impact of temporal changes in the algae may be limited by the dynamics of substrate depletion along the gut following feeding, i.e. the depletion of storage metabolites in the proximal hindgut. Estimated bacterial density, showed that Bacteroidota has the highest density (copies/mL) in distal-most lumen section V, where SCFA concentrations are highest. Bacteroidota genera Alistipes and Rikenella may play important roles in the breakdown of seaweed into useful compounds for the fish host.

Chakrabartyella piscis gen. nov., sp. nov., a member of the family Lachnospiraceae, isolated from the hindgut of the marine herbivorous fish Kyphosus sydneyanus.

A Gram-stain-negative, non-spore-forming, rod-shaped, obligately anaerobic bacterium, designated strain BP5GT, was isolated from the hindgut of a silver drummer (Kyphosus sydneyanus) fish collected from the Hauraki Gulf, New Zealand. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that the isolate belonged to the family Lachnospiraceae in the phylum Bacillota and was most closely related to Anaerotignum propionicum with 94.06 % sequence identity. Isolate BP5GT grew on agar medium containing mannitol and fish gut fluid as carbon sources. Clear colonies of approximately 1 mm diameter of the isolate grew within a week at 20-28 °C (optimum, 28 °C) and pH 7.6-8.5 (optimum, pH 8.5). Strain BP5GT was very sensitive to NaCl and the optimal concentration for growth was 0.045 % (w/v). Acetate and propionate were the major fermentation products. The major cellular fatty acids were C12 : 0, C14 : 0, C15 : 0 and C16 : 0. The genome sequence of the isolate was determined. Its G+C content was 38.41 mol% and the 71.41 % average nucleotide identity of the BP5GT genome to its closest neighbour with a sequenced genome (A. propionicum DSM 1682T) indicated low genomic relatedness. Based on the phenotypic and taxonomic characteristics observed in this study, a novel genus and species named Chakrabartyella piscis gen. nov., sp. nov. is proposed for isolate BP5GT (=ICMP 24687T=JCM 35769T).

Distinct microbiota composition and fermentation products indicate functional compartmentalization in the hindgut of a marine herbivorous fish.

Many marine herbivorous fishes harbour diverse microbial communities in the hindgut that can play important roles in host health and nutrition. Kyphosus sydneyanus is a temperate marine herbivorous fish that feeds predominantly on brown seaweeds. We employed 16S rRNA gene amplicon sequencing and gas chromatography to characterize microbial communities and their metabolites in different hindgut regions of six K. sydneyanus. Measurements were confined to three distal sections of the intestine, labelled III, IV and V from anterior to posterior. A total of 625 operational taxonomic units from 20 phyla and 123 genera were obtained. Bacteroidota, Firmicutes and Proteobacteria were the major phyla in mean relative abundance, which varied along the gut. Firmicutes (76%) was the most dominant group in section III, whereas Bacteroidota (69.3%) dominated section V. Total short-chain fatty acid (SCFA) concentration was highest in sections IV and V, confirming active fermentation in these two most distal sections. The abundance of Bacteroidota correlated with propionate concentration in section V, while Firmicutes positively correlated with formate in sections III and IV. Acetate levels were highest in sections IV and V, which correlated with abundance of Bacteroidota. Despite differences in gut microbial community composition, SCFA profiles were consistent between individual fish in the different hindgut regions of K. sydneyanus, although proportions of SCFAs differed among gut sections. These findings demonstrate functional compartmentalization of the hindgut microbial community, highlighting the need for regional sampling when interpreting overall microbiome function. These results support previous work suggesting that hindgut microbiota in marine herbivorous fish are important to nutrition in some host species by converting dietary carbohydrates into metabolically useful SCFAs.

Tannockella kyphosi gen. nov., sp. nov., a member of the family Erysipelotrichaceae, isolated from the hindgut of the marine herbivorous fish Kyphosus sydneyanus.

A Gram-stain-positive, non-spore-forming, rod-shaped, obligately anaerobic bacterium, designated strain BP52GT, was isolated from the hindgut of a Silver Drummer (Kyphosus sydneyanus) fish collected from the Hauraki Gulf, New Zealand. Phylogenetic analysis based on 16S rRNA gene sequencing indicated that the isolate belonged to the family Erysipelotrichaceae in the phylum Firmicutes and was most closely related to Clostridium saccharogumia with 93.3 % sequence identity. Isolate BP52GT grew on agar medium containing mannitol as the sole carbon source. White, opaque and shiny colonies of the isolate measuring approximately 1 mm diameter grew within a week at 20-28 °C (optimum, 24 °C) and pH 6.9-8.5 (optimum, pH 7.8). BP52GT tolerated the addition of up to 1 % NaCl to the medium. Formate and acetate were the major fermentation products. The major cellular fatty acids were C16 : 0, C16:1n-7t and C18:1n-7t. The genome sequence of the isolate was determined. Its G+C content was 30.7 mol%, and the 72.65 % average nucleotide identity of the BP52GT genome to its closest neighbour with a completely sequenced genome (Erysipelatoclostridium ramosum JCM 1298T) indicated low genomic relatedness. Based on the phenotypic and taxonomic characteristics observed in this study, a novel genus and species Tannockella kyphosi gen. nov., sp. nov. is proposed for isolate BP52GT (=NZRM 4757T=JCM 34692T).

Abalone under moderate heat stress have elevated metabolic rates and changes to digestive enzyme activities.

Abalone around the world are subject to increasing frequency of marine heatwaves, yet we have a limited understanding of how acute high temperature events impact the physiology of these commercially and ecologically important species. This study examines the impact of a 5 °C temperature increase over ambient conditions for six weeks on the metabolic rates, digestive enzyme activities in the digestive gland, and digestive efficiency of Red Abalone (Haliotis rufescens) and Paua (H. iris) on their natural diets. We test the hypothesis that abalone digestive function can keep pace with this increased metabolic demand in two separate experiments, one for each species. H. iris had higher food intake in the heat treatment. Both species had higher metabolic rates in the heat treatment with Q10 = 1.73 and Q10 = 2.46 for H. rufescens and H. iris, respectively. Apparent organic matter digestibility, protein digestibility, and carbohydrate digestibility did not differ between the heat treatment and the ambient (control) treatment in either experiment. H. rufescens exhibited higher maltase, alanine-aminopeptidase, and leucine-aminopeptidase activities in the heat treatment. Amylase, ß-glucosidase, trypsin, and alkaline phosphatase activities in the digestive gland tissue did not differ between temperature treatments. H. iris exhibited lower amylase and ß-glucosidase activities in the heat treatment, while maltase, trypsin, leucine-aminopeptidase, and alkaline phosphatase activities did not differ between treatments. We conclude that over six weeks of moderate heat stress both abalone species were able to maintain digestive function, but achieved this maintenance in species-specific ways.

Substrate degradation pathways, conserved functions and community composition of the hindgut microbiota in the herbivorous marine fish Kyphosus sydneyanus.

Symbiotic gut microbiota in the herbivorous marine fish Kyphosus sydneyanus play an important role in digestion by converting refractory algal carbohydrate into short-chain fatty acids. Here we characterised community composition using both 16S rRNA gene amplicon sequencing and shotgun-metagenome sequencing. Sequencing was carried out on lumen and mucosa samples (radial sections) from three axial sections taken from the hindgut of wild-caught fish. Both lumen and mucosa communities displayed distinct distributions along the hindgut, likely an effect of the differing selection pressures within these hindgut locations, as well as considerable variation among individual fish. In contrast, metagenomic sequences displayed a high level of functional similarity between individual fish and gut sections in the relative abundance of genes (based on sequencing depth) that encoded enzymes involved in algal-derived substrate degradation. These results suggest that the host gut environment selects for functional capacity in symbionts rather than taxonomic identity. Functional annotation of the enzymes encoded by the gut microbiota was carried out to infer the metabolic pathways used by the gut microbiota for the degradation of important dietary substrates: mannitol, alginate, laminarin, fucoidan and galactan (e.g. agar and carrageenan). This work provides the first evidence of the genomic potential of K. sydneyanus hindgut microbiota to convert highly refractory algal carbohydrates into metabolically useful short-chain fatty acids.

Bacillus licheniformis FA6 Affects Zebrafish Lipid Metabolism through Promoting Acetyl-CoA Synthesis and Inhibiting ß-Oxidation.

The intestinal microbiota contributes to energy metabolism, but the molecular mechanisms involved remain less clear. Bacteria of the genus Bacillus regulate lipid metabolism in the host and are thus commonly used as beneficial probiotic supplements. In the present study, Bacillus licheniformis FA6 was selected to assess its role in modulating lipid metabolism of zebrafish (Danio rerio). Combining 16S rRNA high-throughput sequencing, micro-CT scan, metabolic parameters measurement, and gene expression analysis, we demonstrated that B. licheniformis FA6 changed the gut microbiota composition of zebrafish and increased both the Firmicutes/Bacteroidetes ratio and lipid accumulation. In terms of metabolites, B. licheniformis FA6 appeared to promote acetate production, which increased acetyl-CoA levels and promoted lipid synthesis in the liver. In contrast, addition of B. licheniformis lowered carnitine levels, which in turn reduced fatty acid oxidation in the liver. At a molecular level, B. licheniformis FA6 upregulated key genes regulating de novo fatty acid synthesis and downregulated genes encoding key rate-limiting enzymes of fatty acid ß-oxidation, thereby promoting lipid synthesis and reducing fatty acid oxidation. Generally, our results reveal that B. licheniformis FA6 promotes lipid accumulation in zebrafish through improving lipid synthesis and reducing ß-oxidation.

Evolutionary origin of the Atlantic Cabo Verde nibbler (Girella stuebeli), a member of a primarily Pacific Ocean family of antitropical herbivorous reef fishes.

Nibblers (family Girellidae) are reef fishes that are mostly distributed in the Indo-Pacific, with one exception: Girella stuebeli, which is found in the Cabo Verde Archipelago, in the Atlantic Ocean. We capitalized on this unusual distribution to study the evolutionary history of the girellids, and determine the relationship between G. stuebeli and the remaining nibbler taxa. Based on thousands of genomic markers (RAD sequences), we identified the closest relatives of G. stuebeli as being a clade of three species endemic to the northwestern Pacific, restricted to the Sea of Japan and vicinity. This clade diverged from G. stuebeli approximately 2.2 Mya. Two alternative potential routes of migration may explain this affinity: a western route, from the Tropical Eastern Pacific and the Tropical Western Atlantic, and an eastern route via the Indian Ocean and Southern Africa. The geological history and oceanography of the regions combined with molecular data presented here, suggest that the eastern route of invasion (via the Indian Ocean and Southern Africa) is a more likely scenario.

Synchronous biological feedbacks in parrotfishes associated with pantropical coral bleaching.

Biological feedbacks generated through patterns of disturbance are vital for sustaining ecosystem states. Recent ocean warming and thermal anomalies have caused pantropical episodes of coral bleaching, which has led to widespread coral mortality and a range of subsequent effects on coral reef communities. Although the response of many reef-associated fishes to major disturbance events on coral reefs is negative (e.g., reduced abundance and condition), parrotfishes show strong feedbacks after disturbance to living reef structure manifesting as increases in abundance. However, the mechanisms underlying this response are poorly understood. Using biochronological reconstructions of annual otolith (ear stone) growth from two ocean basins, we tested whether parrotfish growth was enhanced following bleaching-related coral mortality, thus providing an organismal mechanism for demographic changes in populations. Both major feeding guilds of parrotfishes (scrapers and excavators) exhibited enhanced growth of individuals after bleaching that was decoupled from expected thermal performance, a pattern that was not evident in other reef fish taxa from the same environment. These results provide evidence for a more nuanced ecological feedback system-one where disturbance plays a key role in mediating parrotfish-benthos interactions. By influencing the biology of assemblages, disturbance can thereby stimulate change in parrotfish grazing intensity and ultimately reef geomorphology over time. This feedback cycle operated historically at within-reef scales; however, our results demonstrate that the scale, magnitude, and severity of recent thermal events are entraining the biological responses of disparate communities to respond in synchrony. This may fundamentally alter feedbacks in the relationships between parrotfishes and reef systems.

World-wide species distributions in the family Kyphosidae (Teleostei: Perciformes).

Sea chubs of the family Kyphosidae are major consumers of macroalgae on both temperate and tropical reefs, where they can comprise a significant proportion of fish biomass. However, the relationships and taxonomic status of sea chubs (including the junior synonyms Hermosilla, Kyphosus, Neoscorpis and Sectator) worldwide have long been problematical due to perceived lack of character differentiation, complicating ecological assessment. More recently, the situation has been further complicated by publication of conflicting taxonomic treatments. Here, we resolve the relationships, taxonomy and distribution of all known species of sea chubs through a combined analysis of partial fragments from mitochondrial markers (12s, 16s, cytb, tRNA -Pro, -Phe, -Thr and -Val) and three nuclear markers (rag1, rag2, tmo4c4). These new results provide independent evidence for the presence of several junior synonyms among Atlantic and Indo-Pacific taxa, demonstrating that several sea chub species are more widespread than previously thought. In particular, our results can reject the hypothesis of endemic species in the Atlantic Ocean. At a higher taxonomic level, our results shed light on the relationships between Girellidae, Kuhliidae, Kyphosidae, Microcanthidae, Oplegnathidae and Scorpididae, with Scorpididae resolved as the sister group to Kyphosidae.

Intestinal microbiota in fishes: what's known and what's not.

High-throughput sequencing approaches have enabled characterizations of the community composition of numerous gut microbial communities, which in turn has enhanced interest in their diversity and functional relationships in different groups of vertebrates. Although fishes represent the greatest taxonomic and ecological diversity of vertebrates, our understanding of their gut microbiota and its functional significance has lagged well behind that of terrestrial vertebrates. In order to highlight emerging issues, we provide an overview of research on fish gut microbiotas and the biology of their hosts. We conclude that microbial community composition must be viewed within an informed context of host ecology and physiology, and that this is of particular importance with respect to research planning and sampling design.

Temperature-related variation in growth rate, size, maturation and life span in a marine herbivorous fish over a latitudinal gradient.

In ectotherms, growth rate, body size and maturation rate covary with temperature, with the direction and magnitude of variation predicted by the Temperature-Size Rule (TSR). Nutritional quality or availability of food, however, may vary over latitudinal gradients, resulting in ambiguous effects on body size and maturation rate. The Temperature-Constraint Hypothesis (TCH) predicts that marine herbivorous ectotherms are nutritionally compromised at latitudes exceeding 30°. This provides an opportunity to resolve the contrasting demographic responses of ectotherms to variation in temperature and nutritional status over latitudinal gradients. This study uses analysis of demographic rates to evaluate the predictions of the TSR in a marine herbivorous ectotherm sampled over a significant latitudinal gradient. The direction and magnitude of demographic variation was established in the marine herbivorous fish, Odax pullus (the butterfish), and compared with that of a phylogenetically related but trophically distinct species, the carnivorous Notolabrus fucicola (the banded wrasse). Both species were sampled at three locations across the length of New Zealand covering latitudes between 35°S and 49°S. Growth rate, mean size-at-age, age- and size-at-maturity, life span and abundance were estimated for each species at each location. Demographic traits of both taxa varied with latitude. Both species showed slower initial growth rates, and matured later at a larger body size at higher latitudes than populations sampled at lower latitudes. In addition, abundances increased significantly at higher latitudes in both species. These results were consistent with the TSR but not with the TCH, confirming that nutritional ecology (herbivore vs. carnivory) did not determine demographic patterns over a biologically significant latitudinal gradient. Results from this study suggest that the absence of herbivorous reef fishes from the higher latitudes of the Northern Hemisphere may not reflect a general physiological mechanism as suggested by the TCH and highlights the need to clarify the evolutionary histories of the marine biota of each hemisphere.

New observations on the ciliate genus Vestibulongum (Pycnotrichidae): vestibular ultrastructure, macronuclear endosymbiotic bacteria, biogeography, and evidence for host specificity.

Two isolates of the pycnotrichid ciliate genus, Vestibulongum, were collected from the host fish, Acanthurus xanthopterus, from two locations in the Southern Pacific Ocean. One was from the Great Barrier Reef (GBR), and a second from Papua New Guinea. These sites are thousands of km from the type locality, off the coast of South Africa. New data were collected from protargol-stained samples to more fully characterize the general form and light microscopic structures of the ciliate. Specimens from all three sites had a long vestibule, characteristic of most members of the family. Data suggest that specimens from each site are the same genus. The kinetids of the Vestibulongum isolated from the GBR contained the typical components of postciliary, transverse, and nemodesmatal microtubules, and Kd fibrils. Also, two quite different forms of endomacronuclear bacteria were observed and are described. One of those has distinct endospores, which are similar to endospores in nuclear endosymbiotic bacteria in a species of Balantidium from the gut of another species of surgeonfish.

Kyphosus gladius, a new species of sea chub from Western Australia (Teleostei: Kyphosidae), with comments on Segutilum klunzingeri Whitley.

Two morphologically distinct forms of the nominal species Kyphosus sydneyanus (Günther, 1886) (Kyphosidae) were discerned while collecting off Western Australia near Perth in 2009. A morphological comparison with recognized species of Kyphosus and an analysis of mtDNA (Cytochrome b, control region, 12S and 16S) and three nDNA markers (RAG1, RAG2 and Tmo-4C4) demonstrated that the more elongate of these forms was an undescribed species of Kyphosus. It differs from congeners in the Pacific and Indian Oceans in the combination of the following characters: green bar on the operculum, 11-12 dorsal soft fin rays, depth of caudal peduncle 9.9-11.8 % SL, body depth 33.3-41.6 % SL, 55-63 scales in lateral line, 12-16 interorbital scales, 44-55 pored scales in the lateral line, 3-5 gill rakers on upper limb of first gill arch internally, 11-15 gill rakers on lower limb of first gill arch internally, 15-19 total gill rakers on first gill arch, and by having 10 precaudal vertebrae and 16 caudal vertebrae. Examination of museum specimens and available underwater photographs suggests that the new species is restricted to Western Australia, and to date it has been recorded between the Houtman Abrolhos Islands and Albany. Discrepancies between the type specimen and original description of Segutilum klunzingeri Whitley made it impossible to determine the relationship between this taxon and the new species from Western Australia, and thus we consider S. klunzingeri a nomen dubium.

The genomic basis for the evolution of a novel form of cellular reproduction in the bacterium Epulopiscium.

BACKGROUND: Epulopiscium sp. type B, a large intestinal bacterial symbiont of the surgeonfish Naso tonganus, does not reproduce by binary fission. Instead, it forms multiple intracellular offspring using a process with morphological features similar to the survival strategy of endospore formation in other Firmicutes. We hypothesize that intracellular offspring formation in Epulopiscium evolved from endospore formation and these two developmental programs share molecular mechanisms that are responsible for the observed morphological similarities. RESULTS: To test this, we sequenced the genome of Epulopiscium sp. type B to draft quality. Comparative analysis with the complete genome of its close, endospore-forming relative, Cellulosilyticum lentocellum, identified homologs of well-known sporulation genes characterized in Bacillus subtilis. Of the 147 highly conserved B. subtilis sporulation genes used in this analysis, we found 57 homologs in the Epulopiscium genome and 87 homologs in the C. lentocellum genome. CONCLUSIONS: Genes coding for components of the central regulatory network which govern the expression of forespore and mother-cell-specific sporulation genes and the machinery used for engulfment appear best conserved. Low conservation of genes expressed late in endospore formation, particularly those that confer resistance properties and encode germinant receptors, suggest that Epulopiscium has lost the ability to form a mature spore. Our findings provide a framework for understanding the evolution of a novel form of cellular reproduction.

Histology and ultrastructure of the gastrointestinal tract in four temperate marine herbivorous fishes.

While alimentary tract anatomy in many terrestrial herbivorous vertebrates is well documented, the digestive systems of marine herbivorous fishes are poorly characterised. The gastrointestinal tract (GIT) of four species of marine herbivorous fishes from northeastern New Zealand, butterfish Odax pullus (Labridae), marblefish Aplodactylus arctidens (Aplodactylidae), notch-head marblefish A. etheridgii (Aplodactylidae) and silver drummer Kyphosus sydneyanus (Kyphosidae), were examined using histology and transmission electron microscopy (TEM) to provide a detailed histological and ultrastructural description of gut anatomy. Gastric glands were distributed over rugae in the stomach of A. arctidens, A. etheridgii and K. sydneyanus. The luminal surface of the stomach of these three species was lined by columnar mucous cells, and oxynticopeptic cells lined the glands in the stomach. Villi were present along the length of the intestine in all four species. The anterior intestine had thin musculature, and was lined by absorptive cells with long microvilli and numerous small vesicles in the apical cytoplasm. The posterior intestine was lined by absorptive columnar cells with long microvilli, invaginations between microvilli with electron-dense membranes, and pinocytotic vesicles. Surface area generally decreased from the anterior to posterior intestine. Histological and ultrastructural results were consistent with lipid absorption occurring in the anterior GIT and protein absorption in the posterior GIT. The results of this study indicate clear differences in GIT structure among the study species, and digestion models based on chemical reactor theory were developed to characterise these differences.

Novel field observations of coral reef fishes feeding on epiphytic and epizoic organisms associated with the allelopathic seaweed Galaxaura divaricata.

In degraded coral reef ecosystems, allelopathic macroalgae have received increasing attention from marine ecologists because their secondary metabolites (also known as allelochemicals) kill corals that grow adjacent to them and weaken the recovery of degraded reefs. One well-known coral-killing macroalga is the calcareous red seaweed Galaxaura. However, our knowledge of how coral reef fishes interact with allelopathic algae like Galaxaura is very limited. Here, we documented novel observations of feeding interactions of 17 species of coral reef fishes (herbivorous and carnivorous) with the filamentous Galaxaura divaricata on degraded lagoon patch reefs in Dongsha Atoll (South China Sea). Video analyses showed that territorial farming damselfishes (i.e., Dischistodus perspicillatus, D. prosopotaenia, Hemiglyphidodon plagiometopon, Pomacentrus grammorhynchus, P. adelus, and Neoglyphidodon nigroris) and juvenile parrotfishes (Scarus schlegeli, S. ghobban, S. rivulatus, and Chlorurus spilurus) likely used G. divaricata as a feeding substratum. Further, microscopic analyses revealed that the filamentous surface of G. divaricata harbored a wealth of epiphytic microalgae, such as filamentous cyanobacteria (i.e., Leptolyngbya, Lyngbya, Rivularia, Oscillatoria, and Stigonema), diatoms (i.e., Synedra, Nitzschia, Mastogloia, and Pleurosigma), and filamentous red algae (i.e., Heterosiphonia), suggesting that these fishes targeted the nutrient-rich microscopic epiphytes rather than the nutrient-poor host. Juvenile benthic carnivores (i.e., Labridae, Parupeneus multifasciatus, and Meiacanthus grammistes) form feeding assemblages with roving parrotfishes to feed on small invertebrates (i.e., amphipods, copepods, isopods, gastropods, and polychaetes) associated with G. divaricata. Given that coral reef fishes appear to target the epiphytes associated with Galaxaura rather than the alga itself, these observations thus substantiate the threat posed by the overgrowth of G. divaricata to coral recovery in degraded reef systems due to the lack of natural grazers.

The spoIIE homolog of Epulopiscium sp. type B is expressed early in intracellular offspring development.

Epulopiscium sp. type B is an enormous intestinal symbiont of the surgeonfish Naso tonganus. Intracellular offspring production in Epulopiscium shares features with endospore formation. Here, we characterize the spoIIE homolog in Epulopiscium. The timing of spoIIE gene expression and presence of interacting partners suggest that the activation of σ(F) occurs early in Epulopiscium offspring development.

Extreme polyploidy in a large bacterium.

Cells rely on diffusion to move metabolites and biomolecules. Diffusion is highly efficient but only over short distances. Although eukaryotic cells have broken free of diffusion-dictated constraints on cell size, most bacteria and archaea are forced to remain small. Exceptions to this rule are found among the bacterial symbionts of surgeonfish; Epulopiscium spp. are cigar-shaped cells that reach lengths in excess of 600 mum. A large Epulopiscium contains thousands of times more DNA than a bacterium such as Escherichia coli, but the composition of this DNA is not well understood. Here, we present evidence that Epulopiscium contains tens of thousands of copies of its genome. Using quantitative, single-cell PCR assays targeting single-copy genes, we have determined that copy number is positively correlated with Epulopiscium cell size. Although other bacteria are known to possess multiple genomes, polyploidy of the magnitude observed in Epulopiscium is unprecedented. The arrangement of genomes around the cell periphery may permit regional responses to local stimuli, thus allowing Epulopiscium to maintain its unusually large size. Surveys of the sequences of single-copy genes (dnaA, recA, and ftsZ) revealed genetic homogeneity within a cell consistent with only a small amount ( approximately 1%) of the parental DNA being transferred to the next generation. The results also suggest that the abundance of genome copies in Epulopiscium may allow for an unstable genetic feature, a long mononucleotide tract, in an essential gene. With the evolution of extreme polyploidy and large cell size, Epulopiscium has acquired some of the advantages of eukaryotic cells.

A new species of deep-water triplefin (Pisces: Tripterygiidae) in the genus Ruanoho from coastal New Zealand waters.

A new endemic species of triplefin Ruanoho scurra is described from deep water (108216 m) on the shelf region around coastal New Zealand (Northland to Stewart Island). It is differentiated from its congeners by the combination of fresh colour (bright yellow spots on the head and anterior body, oblique lines on the dorsal and anal fins, and sub-vertical lines on the caudal) as well as some proportional measurements. Comments are made on the relationship with its congeners, and evolutionary history of the family in New Zealand waters, along with observations on the habitat in which this new species is found. This paper formally describes the species first mentioned in Stewart Clements 2015:1523 as the polkadot triplefin.