Genetic relationships of brook lamprey of the genus <Emphasis Type="Italic">Lampetra</Emphasis> in a Pyrenean stream in Spain

The genetic relationships of a population of brook lamprey Lampetra planeri from Spain were analysed using a fragment of the mitochondrial non-coding region in order to describe its relationships with other European Lampetra populations. DNA sequences were obtained from ten larvae, all corresponding to a single private haplotype. Its closest haplotype differed by five mutations and was found in several samples of Lampetra fluviatlis, both from the North Sea and the Baltic. These results argue for the great conservation value of this brook lamprey population, likely originated from L. fluviatilis ancestors.     

Phylogeography of the European brook lamprey (Lampetra planeri) and the European river lamprey (Lampetra fluviatilis) species pair based on mitochondrial data

The European river lamprey Lampetra fluviatilis and the European brook lamprey Lampetra planeri (Block 1784) are classified as a paired species, characterized by notably different life histories but morphological similarities. Previous work has further shown limited genetic differentiation between these two species at the mitochondrial DNA level. Here, we expand on this previous work, which focused on lamprey species from the Iberian Peninsula in the south and mainland Europe in the north, by sequencing three mitochondrial marker regions of Lampetra individuals from five river systems in Ireland and five in southern Italy. Our results corroborate the previously identified pattern of genetic diversity for the species pair. We also show significant genetic differentiation between Irish and mainland European lamprey populations, suggesting another ichthyogeographic district distinct from those previously defined. Finally, our results stress the importance of southern Italian L. planeri populations, which maintain several private alleles and notable genetic diversity.     

Growth and annual survival estimates to examine the ecology of larval lamprey and the implications of ageing error in fitting models

This study used existing western brook lamprey Lampetra richardsoni age information to fit three different growth models (i.e. von Bertalanffy, Gompertz and logistic) with and without error in age estimates. Among these growth models, there was greater support for the logistic and Gompertz models than the von Bertalanffy model, regardless of ageing error assumptions. The von Bertalanffy model, however, appeared to fit the data well enough to permit survival estimates; using length‐based estimators, annual survival varied between 0·64 (95% credibility interval: 0·44–0·79) and 0·81 (0·79–0·83) depending on ageing and growth process error structure. These estimates are applicable to conservation and management of L. richardsoni and other western lampreys (e.g. Pacific lamprey Entosphenus tridentatus) and can potentially be used in the development of life‐cycle models for these species. These results also suggest that estimators derived from von Bertalanffy growth models should be interpreted with caution if there is high uncertainty in age estimates.     

Isolation and characterization of eight microsatellite loci in the brook lamprey Lampetra planeri (Petromyzontiformes) using 454 sequence data

Eight polymorphic microsatellite loci were developed for the brook lamprey Lampetra planeri through 454 sequencing and their usefulness was tested in 45 individuals of both L. planeri and the river lamprey Lampetra fluviatilis. The number of alleles per loci ranged between two and five; the Italian and Irish populations had a mean expected heterozygosity of 0·388 and 0·424 and a mean observed heterozygosity of 0·418 and 0·411, respectively.     

The ecology of lampreys (Petromyzonidae) in the Loch Lomond area

All three lamprey species occurring in western Europe are found in the Loch Lomond area: Sea Lamprey Petromyzon marinus L., River Lamprey Lampetra fluviatilis (L.) and Brook Lamprey Lampetra planeri (Bloch). There are major differences in ecology among the three species and in each some features seem to be unique to the Lomond populations. Available data on the ecology of the three species in the area are presented and existing threats reviewed. Future research and conservation requirements are outlined.     

Mitochondrial DNA variation of an isolated population of the Adriatic brook lamprey Lampetra zanandreai (Agnatha: Petromyzontidae): phylogeographic and phylogenetic inferences

Two mitochondrial genes were examined to compare an isolated population of the Adriatic brook lamprey Lampetra zanandreai in central Italy with other populations in the species range (Po plain) and with parasitic and freshwater lampreys. A single haplotype, identical to one in a Venetian sample, was found in 10 individuals from the isolated population. The reduced variability is consistent with a history of dispersal after the Pleistocene expansion of the Po basin. The results support the hypothesis of an origin of L. zanandreai and L. fluviatilis–L. planeri from a common anadromous ancestor.     

Using visible implant elastomer to study ammocoete populations with Cormack–Jolly–Seber models

This study examined the efficacy of marking wild populations of lampreys with visible implant elastomer (VIE) for 6–18 months to examine ammocoete movements using Cormack–Jolly–Seber (CJS) open‐population models. These methods were tested on two lamprey populations in different river systems. American brook lamprey Lethenteron appendix at Dyke Creek apparent survival (φ) was high in the summer and winter (c. 0·7), but declined after flow events in the spring and autumn. Sea lamprey Petromyzon marinus at Oquaga Creek φ in the top‐ranked models varied with stream location and time. Estimates of φ were similar to Dyke Creek during the summer (c. 0·7), but declined after flow events and remained low (c. 0·1) in winter. Open‐population models support current understanding of ammocoete movement, i.e. dispersal is driven by high‐flow events at certain times of the year. The present study provides a framework to study ammocoetes with VIE.     

Structure and Conservation of Tandem Repeats in the Mitochondrial DNA Control Region of the Least Brook Lamprey (<Emphasis Type="Italic">Lampetra aepyptera</Emphasis>)

Tandemly repeated sequences are a common feature of vertebrate mitochondrial DNA control regions. However, questions still remain about their mode of evolution, function, and phylogenetic distribution. We report phylogenetic and geographic patterns of variation of control region repeat sequence and number in a nonparasitic lamprey, Lampetra aepyptera. A survey of populations from throughout the species’ range revealed remarkably low repeat sequence polymorphism but some interpopulation variation in repeat number. The high sequence similarity extended to repeats observed in other species in the genus Lampetra and other lamprey genera. The very low levels of variation suggest a high copy turnover. Our data are consistent with the illegitimate elongation model of repeat gain and loss and further suggest that repeat change occurs at internal copies. However, the limited variation across some species of lamprey suggests that functional constraints may further limit variation.     

Basin-scale Patterns in the Drift of Embryonic and Larval Fishes and Lamprey Ammocoetes in two Coastal Rivers

We studied the distribution and abundance of drifting embryonic and larval fishes and lampreys in the Smith and Van Duzen rivers of northern California, U.S.A. We collected seven fish species in four families and at least one lamprey species in the drift. All taxa drifted almost exclusively at night. Sculpins, Cottus aleuticus and C. asper, outnumbered all other taxa, comprising 63% of the catch in the Van Duzen River and 90% of the catch in the Smith River. We estimated that sculpin drift reached 3×10⁷ individualsh^–1 during the relatively high flow period from late winter through early summer. Most sculpin in these two rivers appeared to drift to the estuaries; we estimated 2.5×10⁹ sculpin embryos and larvae reached the Smith River estuary in 1995. In contrast to the sculpins, the patterns in the drift of other taxa suggest limited transport to the estuaries. Suckers, Catostomus occidentalis in the Van Duzen River, C. rimiculus in the Smith River, threespine stickleback, Gasterosteus aculeatus, and lamprey, Lampetra tridentata and possibly L. richardsoni, drifted at much lower rates and later in the year than sculpins. In the Van Duzen River, drift appeared to serve as a dispersal mechanism for only one of three introduced cyprinids. California roach, Lavinia symmetricus, drifted at low rates throughout the summer while we captured only seven Sacramento pikeminnow, Ptychocheilus grandis, and no speckled dace, Rhynichthys osculus. The information gathered on the drift of early life history phases is germane to both the conservation of native fishes and management of non-indigenous species in coastal rivers.     

Contrasting population genetic structure among freshwater‐resident and anadromous lampreys: the role of demographic history,differential dispersal and anthropogenic barriers to movement

The tendency of many species to abandon migration remains a poorly understood aspect of evolutionary biology that may play an important role in promoting species radiation by both allopatric and sympatric mechanisms. Anadromy inherently offers an opportunity for the colonization of freshwater environments, and the shift from an anadromous to a wholly freshwater life history has occurred in many families of fishes. Freshwater‐resident forms have arisen repeatedly among lampreys (within the Petromyzontidae and Mordaciidae), and there has been much debate as to whether anadromous lampreys, and their derived freshwater‐resident analogues, constitute distinct species or are divergent ecotypes of polymorphic species. Samples of 543 European river lamprey Lampetra fluviatilis (mostly from anadromous populations) and freshwater European brook lamprey Lampetra planeri from across 18 sites, primarily in the British Isles, were investigated for 13 polymorphic microsatellite DNA loci, and 108 samples from six of these sites were sequenced for 829 bp of mitochondrial DNA (mtDNA). We found contrasting patterns of population structure for mtDNA and microsatellite DNA markers, such that low diversity and little structure were seen for all populations for mtDNA (consistent with a recent founder expansion event), while fine‐scale structuring was evident for nuclear markers. Strong differentiation for microsatellite DNA loci was seen among freshwater‐resident L. planeri populations and between L. fluviatilis and L. planeri in most cases, but little structure was evident among anadromous L. fluviatilis populations. We conclude that postglacial colonization founded multiple freshwater‐resident populations with strong habitat fidelity and limited dispersal tendencies that became highly differentiated, a pattern that was likely intensified by anthropogenic barriers.     

Alternative migration and host parasitism strategies and their long-term stability in river lampreys from the River Endrick, Scotland

The stability of a discrete body size dimorphism of sexually mature river lamprey Lampetra fluviatilis from the River Endrick, Scotland, was examined over a 21 year period. Stable isotope analysis was used to test the hypothesis that the two size forms comprise individuals with differing migration and parasitic foraging strategies. Maturing river lamprey and the brook lamprey Lampetra planeri were trapped over 3 months each year in the periods 1983–1984 and 2004–2005. Brook lamprey catches and catches of both species combined showed no significant trend in catch rate with time. The catch rate of small body size river lamprey declined between 1983–1984 and 2004–2005 (although the difference did not reach statistical significance; P = 0·055). In contrast, there was a significant increase in the catch rate of the large body size river lamprey and as a consequence, a significant change in the relative proportion of each of the two river lamprey morphs over the study period. Analysis of the stable isotopes of C and N in muscle tissue showed that brook lamprey tissue derived its carbon from a freshwater source and had a δ¹³C more consistent with that of the River Endrick than with Loch Lomond. δ¹⁵N values for this species showed it to be feeding at the base of the food chain, consistent with filter feeding as an ammocoete. The large body size and the small body size river lamprey adults differed substantially in their δ¹³C values, with the small body size δ¹³C signature indicative of a freshwater carbon source and the large body size morph of a marine source. The small body size morph had a δ¹³C signature that was consistent with that of Loch Lomond powan Coregonus lavaretus suggesting that they share a common carbon source. The large body size morph was clearly feeding at a higher trophic level than the small body size morph. A single small body size river lamprey individual with typical morphology for that group, however, had C and N signatures that clustered with those of the large body size morphs. This individual had either migrated to sea to forage, as is typical for the species, or had been feeding on an anadromous fish with a strong marine C signature in fresh water. It is concluded that the body size dimorphism is indicative of a differential migration and foraging strategy in the parasitic phase of the life cycle of river lamprey at this site.     

Morphological abnormalities in a population of Lampetra planeri,with a short review of petromyzontid teratologies

Reported here are several deviations (n = 15; 1·96%) from typical morphology in a large sample (n = 767) of European brook lamprey Lampetra planeri from a single population in the Loch Lomond catchment; this includes one specimen bearing a true anal fin. A brief review of petromyzontid teratology is provided.     

Phylogeographical analysis reveals multiple conservation units in brook lampreys Lampetra planeri of Portuguese streams

The populations of brook lamprey Lampetra planeri of Portuguese Rivers were analysed phylogeographically using a fragment of 644 bp of the mitochondrial control region of 158 individuals from six populations. Samples representing L. planeri and migratory lampreys Lampetra fluviatilis of rivers draining to the North Sea and the Baltic Sea were also included to assess the relationships of Portuguese samples. The data support a clear differentiation of all the populations studied. Several populations, which are isolated among themselves and also from the migratory lampreys, proved to be entirely composed of private haplotypes, a finding that supports some time of independent evolutionary history for these populations. This, combined with the geographic confinement to small water bodies, justifies the recognition of at least four conservation units in the Portuguese rivers Sado, São Pedro, Nabão and Inha.     

Effect of the western pearlshell mussel Margaritifera falcata on Pacific lamprey Lampetra tridentata and ecosystem processes

Suspension feeders concentrate organic material from the water column and enhance deposition to the surrounding benthos. On the South Fork of the Eel River (Mendocino, California) two suspension feeders, the freshwater mussel Margaritifera falcata and Pacific lamprey larvae Lampetra tridentata, co‐occur in areas with low flow velocities and boundary sheer stresses. We investigated mussel/lamprey larvae interactions, and their impacts on nutrient and organic matter cycling, in flow‐through enclosures placed where lamprey larvae and mussels naturally occurred. Over the 80‐day study, lamprey larvae grew faster in the presence of mussels and in food addition treatments. Our results suggest that lamprey larvae benefit from native mussels, and that lamprey populations and organic matter retention in rivers may decrease with the rapid decline of native freshwater mussels.     

Allozyme Polymorphisms and Biochemical-Genetic Taxon Markers in Lampreys (Lampetra, Eudontomyzon, Petromyzon)

Variation at 23 putative enzyme-coding loci was scored in 424 lampreys, including 321 European brook lampreys (Lampetra planeri), 83 European river lampreys (L. fluviatilis), 11 Ukrainian brook lampreys (Eudontomyzon mariae), and nine sea lampreys (Petromyzon marinus). Twelve polymorphisms are described for Lampetra species (LDH*, SOD-2*, PNP*, AAT-1*, AK-1*, ES-2*, LGL*, MPI*, GPI-1*, GPI-2*, PGM*, IDHP-2*), and two each for E. mariae (GPI-1*, ME-2*) and P. marinus (MDH-1*, ME-2*). Diagnostic allozymes are presented for the discrimination of lamprey taxa, some of which are difficult to recognize from the morphology of ammocoetes larvae, the life stage usually encountered when collecting cyclostomes. The allelic markers described permit the clear allocation to a genus, except for the species L. fluviatilis and L. planeri, which are not differentiated by qualitative allozyme analysis.     

HPLC and ELISA analyses of larval bile acids from Pacific and western brook lampreys

Comparative studies were performed on two native lamprey species, Pacific lamprey (Lampetra tridentata) and western brook lamprey (Lampetra richardsoni) from the Pacific coast along with sea lamprey (Petromyzon marinus) from the Great Lakes, to investigate their bile acid production and release. HPLC and ELISA analyses of the gall bladders and liver extract revealed that the major bile acid compound from Pacific and western brook larval lampreys was petromyzonol sulfate (PZS), previously identified as a migratory pheromone in larval sea lamprey. An ELISA for PZS has been developed in a working range of 20 pg-10 ng per well. The tissue concentrations of PZS in gall bladder were 127.40, 145.86, and 276.96 micro g/g body mass in sea lamprey, Pacific lamprey, and western brook lamprey, respectively. Releasing rates for PZS in the three species were measured using ELISA to find that western brook and sea lamprey released PZS 20 times higher than Pacific lamprey did. Further studies are required to determine whether PZS is a chemical cue in Pacific and western brook lampreys.     

Characterization of 12 microsatellite loci for the Pacific lamprey, Entosphenus tridentatus (Petromyzontidae), and cross-amplification in five other lamprey species

The Pacific lamprey (Entosphenus tridentatus) is an anadromous fish that is of conservation concern in North America and Asia. Data on Pacific lamprey population structure are scarce and conflicting, impeding conservation efforts. We optimized 12 polymorphic microsatellite loci for the Pacific lamprey. Three to 13 alleles per locus were observed in a sample of 51 fish collected from the West Fork Illinois River, Oregon. Observed heterozygosity ranged from 0.235 to 0.902 and expected heterozygosity ranged from 0.214 to 0.750. Cross-species amplification produced 8 to 12 polymorphic loci in four other Entosphenus species and in the western brook lamprey (Lampetra richardsoni). Two loci appear to be diagnostic for distinguishing Entosphenus from Lampetra. These markers will be valuable for evaluating population structure and making conservation decisions for E. tridentatus and other lamprey species.     

Using stable isotopes and C:N ratios to examine the life‐history strategies and nutritional sources of larval lampreys

Natural abundance stable‐isotope analysis (δ¹³C and δ¹⁵N) and C:N ratios were used to study the ammocoete phase of two common non‐parasitic lamprey species (least brook lamprey Lampetra aepyptera and American brook lamprey Lethenteron appendix) in two tributaries of the Ohio River (U.S.A.). The C:N ratios suggest that each species employs different lipid accumulation strategies to support its metamorphosis and recruitment into an adult animal. Ammocoete δ¹³C values generally increased with increasing C:N values. In contrast to δ¹³C, ammocoete δ¹⁵N values were weakly related to the total length (L_T) in L. aepyptera, but positively correlated to both L_T and C:N ratios in L. appendix. In L. appendix, C:N also correlated positively with L_T, and presumably age. A Bayesian mixing model using δ¹³C and δ¹⁵N was used to estimate nutritional subsidies of different potential food resources to ammocoetes at each site. The models suggested that although nutritional subsidies to ammocoetes varied as a function of site, ammocoetes were generally reliant on large contributions (42–62% at three sites) from aquatic plants. Contributions from aquatic sediment organic matter were also important at all sites (32–63%) for ammocoetes, with terrestrially derived plant materials contributing smaller amounts (4‐33%). These findings provide important insights into the feeding ecology and nutrition of two species of lampreys. They also suggest that similar and other quantitative approaches are required to (1) fully understand how the observed stable‐isotopes ratios are established in ammocoetes and (2) better assess ammocoete nutritional subsidies in different natal streams.     

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 anadromous 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.     

Population genetics of a cyclostome species pair, river lamprey (Lampetra fluviatilis L.) and brook lamprey (Lampetra planeri Bloch)

Horizontal agarose gel electrophoresis of 24 allozyme loci in four species of Central European lampreys (321 Lampetra planeri , 83 L. fluviatilis , 11 Eudontomyzon mariae and nine Petromyzon marinus ) was used to study the 'paired species' L. fluviatilis and L. planeri . The genetic differentiation of the anadromous river lamprey ( L. fluviatilis ) from the stationary brook lamprey ( L. planeri ) was within the range of ingroup differentiation of the latter, but L. fluviatilis exhibited much greater population cohesion over a more extended geographic range: G _ST = 0.0537 versus G _ST = 0.3398, N _em = 4.402 versus N _em = 0.4856, mean genetic among-stock distances D = 0.0047 versus D = 0.0257. L. planeri populations coexisting geographically with L. fluviatilis in the Rhine and Elbe river systems were genetically more cohesive than L. planeri stocks from the Danubian basin where L. fluviatilis is absent. Danubian L. planeri populations exhibit a lower degree of heterozygosity than brook lampreys from the Rhine river system, but comprise deeper genetic lineages ( G _ST = 0.4629 versus G _ST = 0.2434), despite being sampled from a much more restricted area. Isolation-by-distance is observed for L. planeri from the Danubian but not from the Atlantic drainage basins. Transspecific gene flow between L. planeri from Atlantic drainage basins and the long-distant migrating L. fluviatilis is inferred, raising doubt on the validity of two separate biospecies. E. mariae and P. marinus are clearly differentiated from Lampetra spp. at several allozyme loci.