Recent rapid speciation and ecomorph divergence in Indo‐Australian sea snakes

The viviparous sea snakes (Hydrophiinae) are a young radiation of at least 62 species that display spectacular morphological diversity and high levels of local sympatry. To shed light on the mechanisms underlying sea snake diversification, we investigated recent speciation and eco‐morphological differentiation in a clade of four nominal species with overlapping ranges in Southeast Asia and Australia. Analyses of morphology and stomach contents identified the presence of two distinct ecomorphs: a ‘macrocephalic’ ecomorph that reaches >2 m in length, has a large head and feeds on crevice‐dwelling eels and gobies; and a ‘microcephalic’ ecomorph that rarely exceeds 1 m in length, has a small head and narrow fore‐body and hunts snake eels in burrows. Mitochondrial sequences show a lack of reciprocal monophyly between ecomorphs and among putative species. However, individual assignment based on newly developed microsatellites separated co‐distributed specimens into four significantly differentiated clusters corresponding to morphological species designations, indicating limited recent gene flow and progress towards speciation. A coalescent species tree (based on mitochondrial and nuclear sequences) and isolation‐migration model (mitochondrial and microsatellite markers) suggest between one and three transitions between ecomorphs within the last approximately 1.2 million to approximately 840 000 years. In particular, the macrocephalic ‘eastern’ population of Hydrophis cyanocinctus and microcephalic H. melanocephalus appear to have diverged very recently and rapidly, resulting in major phenotypic differences and restriction of gene flow in sympatry. These results highlight the viviparous sea snakes as a promising system for speciation studies in the marine environment.     

Chemical marking of European glass eels Anguilla anguilla with alizarin red S and in combination with strontium: in situ evaluation of short‐term salinity effects on survival and efficient mass‐marking

This study presents a new chemical double‐marking technique for European glass eels Anguilla anguilla by combing alizarin red S (ARS) and strontium chloride hexahydrate (Sr). Marked eels (double marked with ARS and Sr, but also single marked with ARS) were exposed in situ to brackish water (15 g l^?1 artificial sea salt) for 14 days and did not exhibit increased mortalities compared with unmarked eels. Indeed, no mortality occurred in a marked group during the experiments. Moreover, an efficient mass‐marking approach with low handling effort for both single ARS and double ARS‐Sr techniques is described and was proven to be practicable for large‐scale stocking programmes.     

DNA barcoding of deep‐water notacanthiform fishes (Teleostei,Elopomorpha)

Notacanthiformes Goodrich, 1909, is an order of deep‐sea teleost fishes with a leptocephalus larval stage whose biology and systematics are not widely known. The aim of this work was to apply the DNA barcode standard, based on COI sequence variation, to the delimitation of the species of this order, which is composed by the families Halosauridae (halosaurus) and Notacanthidae (spiny eels). The sequence data used for the analyses were obtained from two sources: 71 samples collected during oceanographic surveys in the North Atlantic Ocean (including the rare species Lipogenys gillii Goode & Bean, 1895) and 95 sequences collected from the BOLD data set. The neighbor‐joining analysis of the barcodes was successful in identifying 96% of the specimens, representing 9 of 16 and 9 of 10 of the recognised species of halosaurus and spiny eels, respectively, including all the current genera. The comparison between the Atlantic and the BOLD data sets also flagged the possibility of occurrences of misidentification and cryptic species. A modern molecular tool like the DNA barcoding supports the previously morphological‐based systematics of the order Notacanthiformes and will provide better access to the taxonomic knowledge of these deep‐water fishes.     

Host hybridization as a potential mechanism of lateral symbiont transfer in deep‐sea vesicomyid clams

Deep‐sea vesicomyid clams live in mutualistic symbiosis with chemosynthetic bacteria that are inherited through the maternal germ line. On evolutionary timescales, strictly vertical transmission should lead to cospeciation of host mitochondrial and symbiont lineages; nonetheless, examples of incongruent phylogenies have been reported, suggesting that symbionts are occasionally horizontally transmitted between host species. The current paradigm for vesicomyid clams holds that direct transfers cause host shifts or mixtures of symbionts. An alternative hypothesis suggests that hybridization between host species might explain symbiont transfers. Two clam species, Archivesica gigas and Phreagena soyoae, frequently co‐occur at deep‐sea hydrocarbon seeps in the eastern Pacific Ocean. Although the two species typically host gammaproteobacterial symbiont lineages marked by divergent 16S rRNA phylotypes, we identified a number of clams with the A. gigas mitotype that hosted symbionts with the P. soyoae phylotype. Demographic inference models based on genome‐wide SNP data and three Sanger sequenced gene markers provided evidence that A. gigas and P. soyoae hybridized in the past, supporting the hypothesis that hybridization might be a viable mechanism of interspecific symbiont transfer. These findings provide new perspectives on the evolution of vertically transmitted symbionts and their hosts in deep‐sea chemosynthetic environments.     

Relationships between fish and otolith size of nine deep‐sea fishes from the Andaman and Nicobar waters,North Indian Ocean

The study presents for the first time the otolith morphology of nine species of deep‐sea fishes. This study was based on sampling carried out on‐board FORV Sagar Sampada (Cruise No 349) during March‐April 2016, along the continental margin of the Andaman and Nicobar Islands in the Bay of Bengal using high speed demersal trawl. Unbroken (complete) otoliths from Polymixia fusca Kotthaus, 1970, Neoepinnula orientalis (Gilchrist & von Bonde 1924), Chlorophthalmus nigromarginatus (Kamohara, 1953), Cubiceps baxteri (McCulloch, 1923), Bembrops caudimacula (Steindachner, 1876), Neoscopelus microchir (Matsubara, 1943), Ostracoberyx dorygenis Fowler, 1934, Synagrops japonicus (Döderlein, 1883), and Bathyclupea hoskynii Alcock 1891) were used for this study. Length–weight relationships (LWR) and the regression between otolith size (width, weight, area and perimeter) and fish length (TL) of nine deep‐sea Fishes were considered. Numerical relationships derived from the relationship between otolith size and the fish can be used as predictors to estimate the prey size as well as to understand trophic relations and food web dynamics of these hitherto unexamined deep‐sea ichthyofauna. LWR showed negative allometric otolith growth in five species; four species showed positive allometric growth. Otolith size to fish size (TL) relation is explained by a simple linear regression considering otolith width (OW), otolith weight (OWe), otolith area (OA) and otolith perimeter (OP). Stronger r² values (>.76) indicate robustness, except for Cubiceps baxteri (r² = .65), and give better estimates for the TL of the fish.     

Critical swimming speed of yellow‐ and silver‐phase European eel (Anguilla anguilla,L.)

The prime objective of this study was to evaluate differences between the swimming performance of two distinct life stages of European eels. The critical swimming speed (U_crit) of 29 yellow‐ and 33 silver‐phase eels was evaluated in a swim tunnel. Silver‐phase eels showed a better swimming performance (U_crit = 0.66 ms^?1) than yellow individuals (U_crit = 0.43 ms^?1). Male and female silver eels reached an identical U_crit despite their different sizes, which may be a strategy to increase the synchronization of arrival at the spawning grounds.     

Spatiotemporal expression pattern of an encephalopsin orthologue of the sea urchin Hemicentrotus pulcherrimus during early development,and its potential role in larval vertical migration

We have cloned and studied Hp‐ECPN, an encephalopsin orthologue of the sea urchin Hemicentrotus pulcherrimus. Hp‐ecpn cDNA was produced and found to contain a 1461‐bp open reading frame that encodes 486 amino acids. Accumulation of Hp‐ecpn mRNA and protein expression occurred at the 14 h postfertilization (hpf) swimming blastula stage and thereafter. The Hp‐ECPN protein was N‐glycosylated, and the amino acid sequence was similar to that of vertebrate encephalopsins. Whole‐mount immunohistochemistry revealed the presence of Hp‐ECPN in cells (ECPN cells) that appeared initially around the tip of the archenteron in 20 hpf early gastrulae. By the 54 hpf pluteus stage, ECPN cells had spread through the aboral ectoderm, and, by the eight‐arm pluteus stage, were restricted to the tips of the larval arms and the posterior end of the body. The number of ECPN cells increased under conditions of continuous light, but decreased under continuous dark. Knockdown of Hp‐ecpn mRNA using morpholino antisense oligonucleotides decreased the number of ECPN cells considerably, and inhibited the vertical swimming of the larvae. This suggested that Hp‐ECPN plays a role in photosensitive larval swimming vertical migration. In adult tissues, the ECPN cells were detected exclusively in tube feet.     

Development of the deep‐sea viviparous quill worm Leptoecia vivipara (Hyalinoeciinae,Onuphidae, Annelida)

Development of Leptoecia vivipara, a brooding deep‐sea onuphid polychaete with a circum‐Antarctic distribution, was studied using light microscopy, scanning and transmission electron microscopy, and histology. All specimens examined were brooding viviparous females or juveniles; no male gametes were detected. Anterior segments of juvenile and adult worms bore paired compact ovaries with clusters of vitellogenic oocytes. In adults, the mid‐body region formed a chamber containing up to 12 offspring at different stages of development, from oocyte to 13 chaetigers. Mature oocytes freely floated in the coelomic fluid, while embryos and juveniles were enclosed in peritoneal envelopes. Chaetal replacement in juveniles and the morphology of the provisional maxillae are described. Leptoecia vivipara is argued to be a progenetic species with juvenile‐like external morphology and accelerated sexual maturation. These traits may have arisen as adaptations to epibenthic life in a high‐latitude deep‐sea environment affected by seasonal pulses of organic matter.     

Isolation of alkane‐degrading bacteria from deep‐sea Mediterranean sediments

Aims: To isolate and identify alkane‐degrading bacteria from deep‐sea superficial sediments sampled at a north‐western Mediterranean station. Methods and Results: Sediments from the water/sediment interface at a 2400 m depth were sampled with a multicorer at the ANTARES site off the French Mediterranean coast and were promptly enriched with Maya crude oil as the sole source of carbon and energy. Alkane‐degrading bacteria belonging to the genera Alcanivorax, Pseudomonas, Marinobacter, Rhodococcus and Clavibacter‐like were isolated, indicating that the same groups were potentially involved in hydrocarbon biodegradation in deep sea as in coastal waters. Conclusions: These results confirm that members of Alcanivorax are important obligate alkane degraders in deep‐sea environments and coexist with other degrading bacteria inhabiting the deep‐subsurface sediment of the Mediterranean. Significance and Impact of the Study: The results suggest that the isolates obtained have potential applications in bioremediation strategies in deep‐sea environments and highlight the need to identify specific piezophilic hydrocarbon‐degrading bacteria (HCB) from these environments.     

Up‐and‐down shift in residence depth of slickheads (Alepocephalidae) revealed by otolith stable oxygen isotopic composition

Otolith δ¹⁸O profiles for four slickhead species (Alepocephalidae) suggested that Alepocephalus umbriceps, Talismania okinawensis and Rouleina watasei migrated hundreds of metres to shallower depths during the juvenile to young stages before returning to their original depth or even deeper waters. Xenodermichthys nodulosus gradually shifted residence depth from shallow to deeper water during their life. These migratory patterns indicated that the slickheads examined had allopatric residence depths at different life stages, which might enhance the pelagic survival and growth rates of the juvenile and young fishes.     

Sexual reproduction of the brooding sponge <Emphasis Type="Italic">Rhopaloeides odorabile</Emphasis>

Sexual reproduction of Rhopaloeides odorabile, a common Great Barrier Reef sponge, was quantified using histological sections from samples collected over two reproductive seasons. Rhopaloeides odorabile is viviparous and gonochoristic with a male biased sex ratio (2.5:1). Commencement and cessation of gametogenesis coincides with rising and falling sea surface temperatures (≈24–29°C). Spermatogenesis occurs from October until January. Females initiate oogenesis in September with the asynchronous development of oocytes, embryos and larvae occurring within the brood chambers. A larval release period of 5–6 weeks occurs during January and February. The minimum size of reproductive individuals was 176 cm³ for females and 192 cm³ for males. The total reproductive output for both sexes shows a positive correlation with size. A relative reproductive output index quantified the maximum reproductive investment at approximately 1 and 3% of the total choanoderm for females and males, respectively, which represents a lower range of reproduction in comparison to other viviparous sponges. Low reproductive output in R. odorabile may be offset by an extended spawning period, alleviating the risk of releasing larvae in potentially adverse conditions via one synchronised spawning event and increasing overall larval survival.     

Large‐scale distribution and activity patterns of an extremely low‐light‐adapted population of green sulfur bacteria in the Black Sea

The Black Sea chemocline represents the largest extant habitat of anoxygenic phototrophic bacteria and harbours a monospecific population of Chlorobium phylotype BS‐1. High‐sensitivity measurements of underwater irradiance and sulfide revealed that the optical properties of the overlying water column were similar across the Black Sea basin, whereas the vertical profiles of sulfide varied strongly between sampling sites and caused a dome‐shaped three‐dimensional distribution of the green sulfur bacteria. In the centres of the western and eastern basins the population of BS‐1 reached upward to depths of 80 and 95 m, respectively, but were detected only at 145 m depth close to the shelf. Using highly concentrated chemocline samples from the centres of the western and eastern basins, the cells were found to be capable of anoxygenic photosynthesis under in situ light conditions and exhibited a photosynthesis–irradiance curve similar to low‐light‐adapted laboratory cultures of Chlorobium BS‐1. Application of a highly specific RT‐qPCR method which targets the internal transcribed spacer (ITS) region of the rrn operon of BS‐1 demonstrated that only cells at the central station are physiologically active in contrast to those at the Black Sea periphery. Based on the detection of ITS‐DNA sequences in the flocculent surface layer of deep‐sea sediments across the Black Sea, the population of BS‐1 has occupied the major part of the basin for the last decade. The continued presence of intact but non‐growing BS‐1 cells at the periphery of the Black Sea indicates that the cells can survive long‐distant transport and exhibit unusually low maintenance energy requirements. According to laboratory measurements, Chlorobium BS‐1 has a maintenance energy requirement of ～1.6–4.9·10^?15 kJ cell^?1 day^?1 which is the lowest value determined for any bacterial culture so far. Chlorobium BS‐1 thus is particularly well adapted to survival under the extreme low‐light conditions of the Black Sea, and can be used as a laboratory model to elucidate general cellular mechanisms of long‐term starvation survival. Because of its adaptation to extreme low‐light marine environments, Chlorobium BS‐1 also represents a suitable indicator for palaeoceanography studies of deep photic zone anoxia in ancient oceans.     

Reconstructing larval growth and habitat use in an amphidromous goby using otolith increments and microchemistry

High‐resolution analysis of growth increments, trace element chemistry and oxygen isotope ratios (δ¹⁸O) in otoliths were combined to assess larval and post‐larval habitat use and growth of Awaous stamineus, an amphidromous goby native to Hawai‘i. Otolith increment widths indicate that all individuals experience a brief period of rapid growth during early life as larvae and that the duration of this growth anomaly is negatively correlated with larval duration. A protracted high‐growth period early in larval life is associated with a lower ratio of Sr:Ca, which may reflect low salinity conditions in nearshore habitats. A distinct shift in δ¹⁸O (range: 4–5‰) is closely associated with the metamorphic mark in otoliths, indicating that larval metamorphosis occurs promptly upon return to fresh water. Strontium and other trace elements are not as tightly coupled to the metamorphosis mark, but confirm the marine‐to‐freshwater transition. Integration of microstructural and microchemical approaches reveals that larvae vary substantially in growth rate, possibly in association with habitat differences. Although time and financial costs make it difficult to achieve large sample sizes, present results show that examining even a small number of individuals can lead to novel inferences about early life history in diadromous fishes and illustrates the value of integrating analyses.     

Length‐weight relationships of five deep‐sea anglerfishes from Andaman and Nicobar waters

Length weight relationships are presented for five deep‐sea anglerfishes collected from Andaman and Nicobar waters during April 2016 using a 38 m high speed demersal trawl II (HSDT II, crustacean version, codend mesh size 40 mm). The b values ranged from 1.80 to 2.76 and the coefficient of variation (r²) ranged from 0.82 to 0.97.     

Length–weight relationships of eight deep‐sea fish species collected from the southwest coast of India

Length‐weight relationships (LWR) for eight deep‐sea fishes belonging to six families collected from the south west coast of India are presented. Specimens were caught by deep‐sea shrimp trawls with mesh size 24 mm during 2011 and 2013. The b values ranged from 2.13 to 3.71 and the coefficient of determination (r²) ranged from 0.84 to 0.98. A new standard length maximum was recorded for Neoscopelus microchir. The b values estimated during the present study were compared with the values estimated using models based on Bayesian approach deposited in Fish Base.     

Location,size and age at onset of metamorphosis in the Japanese eel Anguilla japonica

This study clarifies the location, size and age at the onset of metamorphosis in Japanese eels Anguilla japonica through oceanic surveys, rearing experiments and analyses of the morphology and otoliths of leptocephali and glass eels. Twenty‐eight metamorphosing leptocephali were collected in the mesoscale eddy region to the east of Taiwan during research expeditions in 2004. Rearing experiments showed that the total length (L_T) of leptocephali decreased by an average of 12·5% during metamorphosis and 13·9% during the 2–12 h after death. Thus, the mean back‐calculated L_T at the onset of metamorphosis for 630 glass eels from Taiwan and Japan was estimated at 67·8 ± 2·7 mm (mean ± S.D.). The estimated mean ante‐mortem size of the fully grown pre‐metamorphic leptocephali collected in 2004 was 64·6 ± 3·4 mm, which was consistent with the L_T estimate for glass eels. Otolith analysis showed that the mean age at the onset of metamorphosis was 137 ± 15 days and indicated that Japanese eels may have a recruitment route through the mesoscale eddies to the east of Taiwan in addition to the direct transfer route from the North Equatorial Current to the Kuroshio Current.     

Interspecific and sexual differences in riverine distribution of tropical eels Anguilla spp.

A total of 261 individuals of the four tropical eel species, Anguilla celebesensis, Anguilla marmorata, Anguilla bicolor pacifica and Anguilla interioris, were collected from 12 locations around Sulawesi Island, Indonesia, to gain knowledge about the riverine distribution of tropical eels. Anguilla marmorata was predominant in the lower reaches of Poso River (94·4% of total eel catch in the sampling area), Poso Lake (93·3%), three small inlet rivers of Tomini Bay (100%) and Laa River (92·3%). Anguilla celebesensis occurred frequently in the inlet rivers of Poso Lake (63·5%). Anguilla bicolor pacifica and Anguilla interioris were rare (1.5 and 0.4%, respectively). Otolith Sr:Ca ratio electron‐probe micro analysis (EPMA) for individual migratory histories revealed that 15 A. celebesensis caught in Poso Lake and its inlet rivers were categorized into 14 river eels (Sr:Ca < 2·5) showing upstream migration seemingly at their elver stage and only one sea eel (Sr:Ca ≥ 6·0) that stayed in the marine habitat for the majority of its life after recruiting to Sulawesi Island before its late upstream migration. In A. marmorata, 19 examined eels from Poso Lake and its inlet rivers were all river eels, while 17 eels from the lower reaches of Poso River were two river eels, six sea eels and nine estuarine eels (2·5 ≤ Sr:Ca < 6·0) that mostly lived in the brackish water. The sex ratio of A. celebesensis was highly skewed towards a dominance of females (99%). In A. marmorata, females were predominant in Poso Lake (95·2%), its inlet rivers (94·7%) and Laa River (100%), while males were more frequent in the lower reaches of Poso River (76·5%) and small inlet rivers of Tomini Bay (94·1%). These results indicate that the riverine distribution pattern of tropical eels differs among species and between sexes.     

The Halomonhystera disjuncta population is homogeneous across the Håkon Mosby mud volcano (Barents Sea) but is genetically differentiated from its shallow‐water relatives

The deep sea has a high biodiversity and a characteristic bathyal fauna. Earlier evidence suggested that at least some shallow‐water species invaded the ecosystem followed by radiation leading to endemic deep‐sea lineages with a genetic and/or morphological similarity to their shallow‐water counterparts. The nematode Halomonhystera disjuncta has been reported from shallow‐water habitats and the deep sea [Håkon Mosby mud volcano (HMMV)], but the morphological features and the phylogenetic relationships between deep‐sea and shallow‐water representatives remain largely unknown. Furthermore, nothing is known about the genetic structure of the H. disjuncta population within the HMMV. This study is the first integrative approach in which the morphological and phylogenetic relationships between a deep‐sea and shallow‐water free‐living nematode species are investigated. To elucidate the phylogenetic relationships, we analysed the mitochondrial gene Cytochrome oxidase c subunit I (COI) and three nuclear ribosomal genes (Internal Transcribed Spacer region, 18S and the D2D3 region of 28S). Our results show that deep‐sea nematodes comprise an endemic lineage compared to the shallow‐water representatives with different morphometric features. COI genetic divergence between the deep‐sea and shallow‐water specimens ranges between 19.1% and 25.2%. Taking these findings into account, we conclude that the deep‐sea form is a new species. amova revealed no genetic structure across the HMMV, suggesting that nematodes are able to disperse efficiently in the mud volcano.     

APPLIED ISSUES: Size‐dependent mortality of migratory silver eels at a hydropower plant,and implications for escapement to the sea

1. The European eel population has decreased drastically during recent decades, and new EU‐legislation calls for measures to change this negative trend. This decline has been attributed to a number of factors, including habitat fragmentation by structural barriers that prevent eels moving between freshwater and the sea. The success of downstream migrating adult silver eels migrating past a hydroelectric plant (HEP) in Sweden was examined by radio‐telemetry, and the results were considered in a historical context by analysing catch data from the river for 1957–2006. 2. The choice of routes and passage success were quantified for three treatment groups and one control group of silver eels. The first treatment, the reservoir group (n = 50), was released into the reservoir upstream of the HEP, and these fish could proceed downstream by passing through the HEP (20 mm rack and turbines) or by entering the spill gates into the former channel, bypassing the HEP. The second treatment group (inside rack, n = 15) was released downstream of the 20‐mm rack and had to pass through the turbines to continue migration to the sea. The third treatment group consisted of dead radio‐tagged eels (n = 6) that were released into the turbines to study the extent of drifting by dead individuals. Finally, the control group (n = 50) was released downstream of the HEP to test for effects of confounding factors. 3. Most live individuals displayed migratory behaviour and continued to proceed downstream after release. Only 8% of the fish released in the reservoir or downstream of the HEP (control) did not migrate. The probability of reaching the next HEP, 24 km further downstream, was high for the control group (96%) and the reservoir‐released individuals that passed the HEP via the spill gates and the former channel (83%). Survival was low and size‐dependent for the individuals that passed the turbines (40%) and even lower for the individuals that had to pass through the rack and the turbines (26%). The overall passage success for eels released in the reservoir was 30%, including both routes. 4. Annual catch data from 1957 to 2006 showed that the number of eels in the River Ätran has decreased. Despite this decrease, escapement biomass has remained unchanged, because of the fact that the mean size of eels has doubled. Passage data from 2007 show that changes in size and abundance have resulted in a reduction of relative escapement to the sea to values that are 21–24% of what they were in 1957–66. However, this low level of escapement could potentially be rectified if appropriate measures facilitating HEP passage are successfully implemented, since the potential escapement biomass in the river, owing to the large size of the eels, has changed little since the 1950s.     

Length–weight relationships of 11 deep‐sea fishes from the western Bay of Bengal and Andaman waters,India

Length–weight relationships (LWRs) are presented for 11 deep‐sea fishes caught in the western Bay of Bengal and Andaman waters during August 2010 using a 38 m high speed demersal trawl II (HSDT II, crustacean version, codend mesh size 40 mm) and a 45.6 m Expo model demersal trawl (codend mesh size 30 mm). The b values ranged from 2.34 to 3.3 and the coefficient of variation (r²) ranged from .82 to .98. LWR estimates of eight deep‐sea fishes are provided for the first time. The estimated LWR values were compared with the Bayesian LWR estimates available in FishBase, based on models developed to improve the accuracy and predictability of species‐specific growth parameters of data‐poor species.