Lamprey Species Research Articles

Bioinformatic Characterization of Genes and Proteins Involved in Blood Clotting in Lampreys

Lampreys and hagfish are the earliest diverging of extant vertebrates and are obvious targets for investigating the origins of complex biochemical systems found in mammals. Currently, the simplest approach for such inquiries is to search for the presence of relevant genes in whole genome sequence (WGS) assemblies. Unhappily, in the past a high-quality complete genome sequence has not been available for either lampreys or hagfish, precluding the possibility of proving gene absence. Recently, improved but still incomplete genome assemblies for two species of lamprey have been posted, and, taken together with an extensive collection of short sequences in the NCBI trace archive, they have made it possible to make reliable counts for specific gene families. Particularly, a multi-source tactic has been used to study the lamprey blood clotting system with regard to the presence and absence of genes known to occur in higher vertebrates. As was suggested in earlier studies, lampreys lack genes for coagulation factors VIII and IX, both of which are critical for the "intrinsic" clotting system and responsible for hemophilia in humans. On the other hand, they have three each of genes for factors VII and X, participants in the "extrinsic" clotting system. The strategy of using raw trace sequence "reads" together with partial WGS assemblies for lampreys can be used in studies on the early evolution of other biochemical systems in vertebrates.

Termophilic and Eastern Pacific migrants in ichthyofauna in the Pacific waters off the northern Kuril Islands and Kamchatka in the 20th–21st centuries

The data on the species composition and occurrence of the termophilic and Eastern Pacific representatives of ichthyofauna in the Pacific waters off the northern Kuril Islands and eastern Kamchatka in the 20th–21st centuries are provided. Thirty-three species of termophilic and twelve species of Eastern Pacific fish and lampreys from 35 families have been registered during this period in the Pacific waters near the Kuril Islands and Kamchatka. Annual and seasonal dynamics of appearance of some fish species in the surveyed region in 1992–2002 are analyzed.

Low reproductive isolation and highly variable levels of gene flow reveal limited progress towards speciation between European river and brook lampreys.

Ecologically based divergent selection is a factor that could drive reproductive isolation even in the presence of gene flow. Population pairs arrayed along a continuum of divergence provide a good opportunity to address this issue. Here, we used a combination of mating trials, experimental crosses and population genetic analyses to investigate the evolution of reproductive isolation between two closely related species of lampreys with distinct life histories. We used microsatellite markers to genotype over 1000 individuals of the migratory parasitic river lamprey (Lampetra fluviatilis) and freshwater-resident nonparasitic brook lamprey (Lampetra planeri) distributed in 10 sympatric and parapatric population pairs in France. Mating trials, parentage analyses and artificial fertilizations demonstrated a low level of reproductive isolation between species even though size-assortative mating may contribute to isolation. Most parapatric population pairs were strongly differentiated due to the joint effects of geographic distance and barriers to migration. In contrast, we found variable levels of gene flow between sympatric populations ranging from panmixia to moderate differentiation, which indicates a gradient of divergence with some population pairs that may correspond to alternative morphs or ecotypes of a single species and others that remain partially isolated. Ecologically based divergent selection may explain these variable levels of divergence among sympatric population pairs, but incomplete genome swamping following secondary contact could have also played a role. Overall, this study illustrates how highly differentiated phenotypes can be maintained despite high levels of gene flow that limit the progress towards speciation.

Studies on endangered and rare non-commercial fish species recorded in the Pomeranian Bay (southern Baltic Sea) in 2010–2013

This paper presents the results of studies on endangered and rare non-commercial fish species (Spinachia spinachia, Nerophis ophidion, Syngnathus typhle, Agonus cataphractus, Pholis gunnellus, Enchelyopus cimbrius, Cyclopterus lumpus) and one lamprey species (Lampetra fluviatilis), recorded as bycatch during monitoring surveys in 2010–2013 in the Pomeranian Bay. Two species were observed for the first time in the Pomeranian Bay: A. cataphractus and E. cimbrius. Descriptions of parasite fauna are provided for C. lumpus and E. cimbrius, which were infected with four pathogenic species from Neomonada, Digenea, Nematoda, and Acanthocephala. Almost all parasite species were new in the hosts examined.

Fecundity of the lampreys Lampetra hubbsi and Eudontomyzon hellenicus

This is the first report on the fecundity in adults of two nonparasitic lamprey species at risk, the Vulnerable Kern Brook Lamprey, Lampetra hubbsi and the Critically Endangered Macedonia Brook Lamprey, Eudontomyzon hellenicus. The mean absolute fecundity of L. hubbsi was 1865 eggs and that of E. hellenicus was 994. The higher fecundity in L. hubbsi was directly related to its higher mean gonadal mass (0.90 g) relative to that in E. hellenicus (0.52 g). Based on preserved museum material possessing well-developed primary and secondary sexual characters, the spawning period for L. hubbsi was inferred to be February-March and for E. hellenicus to either extend from January to May or to occur as separate events; one in January and the other in May. Because metamorphosing ammocoetes of E. hellenicus have been collected in October and in January, it is hypothesized that those metamorphosing in October spawn in January and those metamorphosing in January spawn in May. These periods need to be corroborated with field observations. Evidence for E. hellenicus spawning in January is based on the presence of spent females of this species collected in that month.

Molecular prey identification in Central European piscivores.

Diet analysis is an important aspect when investigating the ecology of fish‐eating animals and essential for assessing their functional role in food webs across aquatic and terrestrial ecosystems. The identification of fish remains in dietary samples, however, can be time‐consuming and unsatisfying using conventional morphological analysis of prey remains. Here, we present a two‐step multiplex PCR system, comprised of six assays, allowing for rapid, sensitive and specific detection of fish DNA in dietary samples. This approach encompasses 78 fish and lamprey species native to Central European freshwaters and enables the identification of 31 species, six genera, two families, two orders and two fish family clusters. All targeted taxa were successfully amplified from 25 template molecules, and each assay was specific when tested against a wide range of invertebrates and vertebrates inhabiting aquatic environments. The applicability of the multiplex PCR system was evaluated in a feeding trial, wherein it outperformed morphological prey analysis regarding species‐specific prey identification in faeces of Eurasian otters. Additionally, a wide spectrum of fish species was detected in field‐collected faecal samples and regurgitated pellets of Common Kingfishers and Great Cormorants, demonstrating the broad applicability of the approach. In conclusion, this multiplex PCR system provides an efficient, easy to use and cost‐effective tool for assessing the trophic ecology of piscivores in Central Europe. Furthermore, the multiplex PCRs and the primers described therein will be applicable wherever DNA of the targeted fish species needs to be detected at high sensitivity and specificity.

Evolution of retinoic acid receptors in chordates: insights from three lamprey species, Lampetra fluviatilis, Petromyzon marinus, and Lethenteron japonicum

BackgroundRetinoic acid (RA) signaling controls many developmental processes in chordates, from early axis specification to late organogenesis. The functions of RA are chiefly mediated by a subfamily of nuclear hormone receptors, the retinoic acid receptors (RARs), that act as ligand-activated transcription factors. While RARs have been extensively studied in jawed vertebrates (that is, gnathostomes) and invertebrate chordates, very little is known about the repertoire and developmental roles of RARs in cyclostomes, which are extant jawless vertebrates. Here, we present the first extensive study of cyclostome RARs focusing on three different lamprey species: the European freshwater lamprey, Lampetra fluviatilis, the sea lamprey, Petromyzon marinus, and the Japanese lamprey, Lethenteron japonicum.ResultsWe identified four rar paralogs (rar1, rar2, rar3, and rar4) in each of the three lamprey species, and phylogenetic analyses indicate a complex evolutionary history of lamprey rar genes including the origin of rar1 and rar4 by lineage-specific duplication after the lamprey-hagfish split. We further assessed their expression patterns during embryonic development by in situ hybridization. The results show that lamprey rar genes are generally characterized by dynamic and highly specific expression domains in different embryonic tissues. In particular, lamprey rar genes exhibit combinatorial expression domains in the anterior central nervous system (CNS) and the pharyngeal region.ConclusionsOur results indicate that the genome of lampreys encodes at least four rar genes and suggest that the lamprey rar complement arose from vertebrate-specific whole genome duplications followed by a lamprey-specific duplication event. Moreover, we describe a combinatorial code of lamprey rar expression in both anterior CNS and pharynx resulting from dynamic and highly specific expression patterns during embryonic development. This ‘RAR code’ might function in regionalization and patterning of these two tissues by differentially modulating the expression of downstream effector genes during development.Electronic supplementary materialThe online version of this article (doi:10.1186/s13227-015-0016-4) contains supplementary material, which is available to authorized users.

Characterization of two neurogenin genes from the brook lamprey lampetra planeri and their expression in the lamprey nervous system.

Neurogenins are required for the specification of neuronal precursors and regulate the expression of basic Helix-Loop-Helix genes involved in neuronal differentiation. Jawed vertebrates possess three Neurogenin paralogy groups and their combined expression covers the entire nervous system, apart from the autonomic nervous system. Here we report the isolation of two Neurogenin genes, LpNgnA and LpNgnB, from the lamprey Lampetra planeri. Phylogenetic analyses show both genes have orthologues in other lamprey species and in a hagfish. Neither gene shows evidence of orthology to specific jawed vertebrate Neurogenin paralogues. LpNgnA is expressed in the ventricular zone of regions of the brain and spinal cord, with expression in the brain demarcating brain sub-compartments including the pallium, tegmentum, tectum, and dorsal thalamus. In the peripheral nervous system, LpNgnA is expressed in cranial sensory placodes and their derivatives, and in the dorsal root ganglia. LpNgnB is expressed transiently in placodal head ectoderm and throughout the central nervous system in early development, and in a small population cells that form part of the macula. Combined, LpNgnA and LpNgnB were detected in most cell populations marked by Neurogenin gene expression in jawed vertebrates, with the exception of the cerebellum, retina and the non-neural expression sites. Developmental Dynamics 244:1096-1108, 2015. © 2015 Wiley Periodicals, Inc.

Variations in the presence of chloride cells in the gills of lampreys (Petromyzontiformes) and their evolutionary implications.

Although confined to fresh water, non-parasitic species of lampreys and the landlocked parasitic sea lamprey, all of which were derived relatively recently from an adromous ancestors, still develop chloride cells, whose function in their ancestors was for osmoregulation in marine waters during the adult parasitic phase. In contrast, such cells are not developed by the non-parasitic least brook lamprey Lampetra aepyptera, which has been separated from its ancestor for >2 million years, nor by the freshwater parasitic species of the genus Ichthyomyzon. The length of time that a non-parasitic species or landlocked parasitic form or species has spent in fresh water is thus considered the overriding factor determining whether chloride cells are developed by those lampreys.

Application of a putative alarm cue hastens the arrival of invasive sea lamprey (Petromyzon marinus) at a trapping location

The sea lamprey Petromyzon marinus is an invasive pest in the Laurentian Great Lakes basin, threatening the persistence of important commercial and recreational fisheries. There is substantial interest in developing effective trapping practices via the application of behavior-modifying semiochemicals (odors). Here we report on the effectiveness of utilizing repellent and attractant odors in a push–pull configuration, commonly employed to tackle invertebrate pests, to improve trapping efficacy at permanent barriers to sea lamprey migration. When a half-stream channel was activated by a naturally derived repellent odor (a putative alarm cue), we found that sea lamprey located a trap entrance significantly faster than when no odor was present as a result of their redistribution within the stream. The presence of a partial sex pheromone, acting as an attractant within the trap, was not found to further decrease the time to when sea lamprey located a trap entrance relative to when the alarm cue alone was applied. Neither the application of alarm cue singly nor alarm cue and partial sex pheromone in combination was found to improve the numbers of sea lamprey captured in the trap versus when no odor was present — likely because nominal capture rate during control trials was unusually high during the study period. Behavioural guidance using these odors has the potential to both improve control of invasive non-native sea lamprey in the Great Lakes as well as improving the efficiency of fish passage devices used in the restoration of threatened lamprey species elsewhere.

The globin gene repertoire of lampreys: convergent evolution of hemoglobin and myoglobin in jawed and jawless vertebrates.

Agnathans (jawless vertebrates) occupy a key phylogenetic position for illuminating the evolution of vertebrate anatomy and physiology. Evaluation of the agnathan globin gene repertoire can thus aid efforts to reconstruct the origin and evolution of the globin genes of vertebrates, a superfamily that includes the well-known model proteins hemoglobin and myoglobin. Here, we report a comprehensive analysis of the genome of the sea lamprey (Petromyzon marinus) which revealed 23 intact globin genes and two hemoglobin pseudogenes. Analyses of the genome of the Arctic lamprey (Lethenteron camtschaticum) identified 18 full length and five partial globin gene sequences. The majority of the globin genes in both lamprey species correspond to the known agnathan hemoglobins. Both genomes harbor two copies of globin X, an ancient globin gene that has a broad phylogenetic distribution in the animal kingdom. Surprisingly, we found no evidence for an ortholog of neuroglobin in the lamprey genomes. Expression and phylogenetic analyses identified an ortholog of cytoglobin in the lampreys; in fact, our results indicate that cytoglobin is the only orthologous vertebrate-specific globin that has been retained in both gnathostomes and agnathans. Notably, we also found two globins that are highly expressed in the heart of P. marinus, thus representing functional myoglobins. Both genes have orthologs in L. camtschaticum. Phylogenetic analyses indicate that these heart-expressed globins are not orthologous to the myoglobins of jawed vertebrates (Gnathostomata), but originated independently within the agnathans. The agnathan myoglobin and hemoglobin proteins form a monophyletic group to the exclusion of functionally analogous myoglobins and hemoglobins of gnathostomes, indicating that specialized respiratory proteins for O2 transport in the blood and O2 storage in the striated muscles evolved independently in both lineages. This dual convergence of O2-transport and O2-storage proteins in agnathans and gnathostomes involved the convergent co-option of different precursor proteins in the ancestral globin repertoire of vertebrates.

Egg drift and hatching success in European river lamprey Lampetra fluviatilis: is egg deposition in gravel vital to spawning success?

Abstract The European river lamprey Lampetra fluviatilis (Linnaeus, 1758) is a threatened species, formerly widespread throughout western Europe, for which loss and degradation of habitat is one of the main causes of decline. As with other lamprey species, areas of gravel substrate with moderate flows are considered necessary for spawning and egg development of river lamprey. This study investigated the dispersal of river lamprey eggs downstream of a spawning area and the hatching success of eggs in the laboratory under a range of potential abiotic conditions (substrate type, water flow and dissolved oxygen level) which eggs could experience in nature. Lamprey eggs were found to drift for a minimum of 50 m downstream of spawning excavations, facilitating dispersal in riffle habitat and to bankside depositional zones. Under conditions mimicking natural microhabitats, but without predation, median egg hatching success was 85.0% in 'spawning habitat' conditions, but surprisingly, was 50.2% in 'larval habitat' conditions with natural silt. The study suggests that egg dispersal out of spawning excavations may be common in this species and demonstrates that habitat located downstream of spawning areas, even larval habitat characterized by fine sediment and moderate to low flow rates, could play an important role in larval recruitment. This suggests that even small areas of gravel or degraded spawning habitat may enable a higher degree of spawning success than has previously been assumed to be necessary for conservation or recolonization of this species. Copyright © 2014 John Wiley & Sons, Ltd.

Use of genotyping by sequencing data to develop a high-throughput and multifunctional SNP panel for conservation applications in Pacific lamprey.

Next-generation sequencing data can be mined for highly informative single nucleotide polymorphisms (SNPs) to develop high-throughput genomic assays for nonmodel organisms. However, choosing a set of SNPs to address a variety of objectives can be difficult because SNPs are often not equally informative. We developed an optimal combination of 96 high-throughput SNP assays from a total of 4439 SNPs identified in a previous study of Pacific lamprey (Entosphenus tridentatus) and used them to address four disparate objectives: parentage analysis, species identification and characterization of neutral and adaptive variation. Nine of these SNPs are FST outliers, and five of these outliers are localized within genes and significantly associated with geography, run-timing and dwarf life history. Two of the 96 SNPs were diagnostic for two other lamprey species that were morphologically indistinguishable at early larval stages and were sympatric in the Pacific Northwest. The majority (85) of SNPs in the panel were highly informative for parentage analysis, that is, putatively neutral with high minor allele frequency across the species' range. Results from three case studies are presented to demonstrate the broad utility of this panel of SNP markers in this species. As Pacific lamprey populations are undergoing rapid decline, these SNPs provide an important resource to address critical uncertainties associated with the conservation and recovery of this imperiled species.

Reduced fecundity in non‐parasitic lampreys may not be due to heterochronic shift in ovarian differentiation

Heterochrony is an important mechanism for the evolution of differences between and within species. In lampreys, heterochrony has been suggested as a mechanism contributing to fecundity differences in non-parasitic versus parasitic species. Non-parasitic lampreys, which do not feed at all after metamorphosis, have much smaller body sizes at maturation and therefore much lower fecundity than parasitic lampreys. Previous studies have suggested that this fecundity difference is established in the larval stage through ovarian differentiation at a younger age (and therefore smaller body size) in non-parasitic females, leading to production of fewer oocytes. The current study examined whether this pattern is applicable in two additional lamprey species. The timing of histological ovarian differentiation was determined in larval parasitic chestnut lamprey Ichthyomyzon castaneus and non-parasitic northern brook lamprey I. fossor in southeastern Manitoba, Canada, and potential fecundity was compared through oocyte counts in differentiated females of both species. Ovarian differentiation occurred in age classes 1 and 2 in both chestnut and northern brook lampreys, and there were no significant differences in the timing of ovarian differentiation between species; this indicates that the timing of ovarian differentiation is not a reliable marker of life history type. Factors such as growth and body condition may determine whether an individual undergoes ovarian differentiation in age class 1 or 2. Chestnut and northern brook lampreys had similar minimum oocyte counts; however, the average and maximum oocyte counts were higher in chestnut lamprey in each age class. As the timing of ovarian differentiation is similar in chestnut and northern brook lampreys, the higher potential fecundity of chestnut lamprey must originate through mechanisms other than heterochrony.

Status assessment of lampreys in Natura 2000 network in Lithuania

The two lamprey species, European river lamprey Lampetra fluviatilis L. and European brook lamprey L. planeri Bloch, which inhabit Lithuanian rivers are of conservation concern. They are considered either Vulnerable, Critically Endangered, or even Extinct in different regions of Europe. Migration obstacles and consequent habitat loss and population fragmentation are probably the most widespread and significant factors contributing to the current status. Therefore, all member states are obliged to designate Special Areas of Conservation (SACs). This study assessed the conservation status of L. fluviatilis and L. planeri in 16 Proposed Sites of Community Interest (pSCIs) for lamprey conservation in Lithuania. Lamprey ammocoete surveys were conducted at 41 sites following the established national protocol. In particular, we compare historical and recent data of lampreys in Lithuanian rivers, discuss the factors that contributed most to their decline and recommend conservation measures that may contribute to their recovery.

Complete genome sequence of the mitochondrial DNA of the river lamprey, Lethenteron japonicum

Lampreys are eel-like jawless fishes evolutionarily positioned between invertebrates and vertebrates, and have been used as model organisms to explore vertebrate evolution. In this study we determined the complete genome sequence of the mitochondrial DNA of the Japanese river lamprey, Lethenteron japonicum, using next-generation sequencers. The sequence was 16,272 bp in length. The gene content and order were identical to those of the sea lamprey, Petromyzon marinus, which has been the reference among lamprey species. However, the sequence similarity was less than 90%, suggesting the need for the whole-genome sequencing of L. japonicum.

A spatial age-structured model for describing sea lamprey (Petromyzon marinus) population dynamics

The control of invasive sea lampreys (Petromyzon marinus) presents large-scale management challenges in the Laurentian Great Lakes. No modeling approach has been developed that describes spatial dynamics of lamprey populations. We developed and validated a spatial and age-structured model and applied it to a sea lamprey population in a large river in the Great Lakes basin. We considered 75 discrete spatial areas, included a stock–recruitment function, spatial recruitment patterns, natural mortality, chemical treatment mortality, and larval metamorphosis. Recruitment was variable, and an upstream shift in recruitment location was observed over time. From 1993 to 2011, recruitment, larval abundance, and the abundance of metamorphosing individuals decreased by 80%, 84%, and 86%, respectively. The model successfully identified areas of high larval abundance and showed that areas of low larval density contribute significantly to the population. Estimated treatment mortality was less than expected but had a large population-level impact. The results and general approach of this work have applications for sea lamprey control throughout the Great Lakes and for the restoration and conservation of native lamprey species globally.

Sneak male mating tactics between lampreys (Petromyzontiformes) exhibiting alternative life‐history strategies

Interspecific sneak male mating tactics between paired lamprey species are described for the first time. Although alternative mating tactics among petromyzontids have been described previously, including intraspecific sneak males, the presence of sneak male tactics between parasitic and non-parasitic forms suggests that high levels of gene flow between putative lamprey species could remain high, despite large body size discrepancies.

Distribution of Pacific LampreyEntosphenus tridentatusin Watersheds of Puget Sound Based on Smolt Monitoring Data

Abstract Lamprey populations are in decline worldwide and the status of Pacific lamprey (Entosphenus tridentatus) is a topic of current interest. They and other lamprey species cycle nutrients and serve as prey in riverine ecosystems. To determine the current distribution of Pacific lamprey in major watersheds flowing into Puget Sound, Washington, we sampled lamprey captured during salmonid smolt monitoring that occurred from late winter to mid-summer. We found Pacific lamprey in 12 of 18 watersheds and they were most common in southern Puget Sound watersheds and in watersheds draining western Puget Sound (Hood Canal). Two additional species, western brook lamprey (Lampetra richardsoni) and river lamprey (L. ayresii) were more common in eastern Puget Sound watersheds. Few Pacific lamprey macrophthalmia were found, suggesting that the majority of juveniles migrated seaward during other time periods. In addition, “dwarf” adult Pacific lamprey (< 300 mm) were observed in several watersheds and may represent ...

Post-zygotic hybrid viability in sympatric species pairs: a case study from European lampreys

Ecological speciation mechanisms are widely assumed to play an important role in the early stages of divergence between incipient species, and this especially true of fishes. In the present study, we tested explicitly for post-zygotic barriers to gene flow between a sympatric, recently diverged lamprey species pair that likely arose through ecological divergence. Experimental in vitro hybridization between anadromous parasitic Lampetra fluviatilis and resident nonparasitic Lampetra planeri resulted in a high proportion of embryos capable of attaining the burrowing pro-larval stage, strongly indicating no post-zygotic barriers to gene flow between these species. A sympatric, locally-adapted resident parasitic form of L. fluviatilis was also found to successfully hybridize with both members of this species pair. The consequences of these findings are discussed in the context of lamprey speciation. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 108, 378–383.