Brook Lamprey Research Articles

Comparative study on a unique architecture of the brook lamprey liver and that of the hagfish and banded houndshark liver.

Although the liver of the lamprey, a group of cyclostomes that diverged the earliest among vertebrates, has abundant bile ducts in the larval stage, which degenerate during metamorphosis, there is no comparative study on its architecture with other early diverged vertebrates in terms of the morphological evolution of vertebrate livers. The present study was undertaken to compare the characteristics of the brook lamprey liver with those of the hagfish and banded houndshark, which have the portal triad type liver architecture, and to discuss its evolution. Although the liver of the brook lamprey had two-cell cords of hepatocytes lined by sinusoids in the ammocoetes larval stage, intrahepatic bile ducts around portal veins penetrated into the liver parenchyma with convolution and gradual reduction in diameter. They also faced dilated sinusoids. The epithelial cells had characteristic intercellular spaces. These characteristics were distinct from those of bile ducts in the hagfish and banded houndshark livers. Although the liver architectures of the hagfish and banded houndshark were similar, the latter penetrated the intrahepatic bile ducts more deeply along the portal veins than the former, in which intrahepatic bile ducts were restricted near the hilum. After metamorphosis, bile ducts degenerated in brook lampreys. These data indicate that the liver architecture of the ammocoetes larva is unique in the parenchymal distribution of bile ducts, their sinusoidal facing, and morphology among extant vertebrates. The periportal distribution of intrahepatic biliary structures may have been established prior to the divergence of the cyclostomes and gnathostomes.

Expanding the known range and practical conservation issues of the Endangered Australian brook lamprey Mordacia praecox

Parasitic and non-parasitic lamprey ‘species pairs’ are an ongoing source of taxonomic uncertainty globally. The geographic range of the only non-parasitic lamprey in the Southern Hemisphere, the Endangered Australian brook lamprey Mordacia praecox, has remained ambiguous since its description in 1968. The conservation of this species is complex as it is presently genetically and, for most of its lifecycle, morphologically indistinguishable from its parasitic paired species, short-headed lamprey M. mordax, the conservation status of which is Least Concern. Difficulty in their identification, coupled with their cryptic behaviour, has resulted in limited knowledge of the species’ ecology and distribution. This is further complicated by the sympatric geographic ranges of the paired species. Using incidental captures, targeted surveys, and open-access wildlife information database records, we describe the discovery and confirmation of lamprey populations from tropical and sub-tropical Australia and the associated ∼1400 km (5-fold) northward extension of the known geographic range of Australian brook lamprey. Surveys yielded non-parasitic lampreys of all life-history stages across 6 tropical and sub-tropical coastal watersheds of eastern Australia. We also highlight major issues associated with the conservation of a cryptic and poorly understood species and discuss potential conservation actions that may, at least in part, ameliorate such issues.

Are lipids always depleted? Comparison of hydrogen, carbon, and nitrogen isotopic values in the muscle and lipid of larval lampreys.

Stable isotope ratios in organisms can be used to estimate dietary source contributions, but lipids must first be accounted for to interpret values meaningfully. Lipids are depleted in heavy isotopes because during lipid synthesis light isotopes of carbon (12C) and hydrogen (1H) are preferentially incorporated. Prior work in larval lampreys has noted unusual lipid effects, which suggest lipids are enriched in the heavy isotope of carbon (13C), but still depleted in the heavy isotope of hydrogen (deuterium; 2H); nitrogen, a relatively rare element in lipids, has not been identified as being as sensitive to lipid content. Our objective was to determine if stable isotope ratios of hydrogen, carbon, and nitrogen behaved as expected in larval lampreys, or if their lipids presented different isotopic behavior. The δ2H, δ13C, and δ15N were measured from the muscle of four lamprey species before and after lipid extraction. In addition, muscle of least brook lamprey (Lampetra aepyptera) was collected every three months for a year from two streams in Maryland. Isotopic ratios were measured in bulk and lipid-extracted muscles, as well as in extracted lipids. The difference between muscle samples before and after lipid extraction (Δδ2H, Δδ13C, Δδ15N) was positively related to lipid proxy (%H or C:N ratio) and were fit best by linear models for Δδ2H and Δδ15N, and by a non-linear model for Δδ13C. The difference between lipid-extracted muscle and lipid δ13C (ΔMLδ13C) was negative and varied between months (ANOVA, F3,53 = 5.05, p < 0.005). Our work suggests that while lipids are often depleted in 13C, this is not a universal rule; however, the depletion of 2H in lipid synthesis appears broadly true.

Phylogenetic and biogeographic history of brook lampreys (Lampetra: Petromyzontidae) in the river basins of the Adriatic Sea based on DNA barcode data.

The Adriatic brook lamprey, Lampetra zanandreai Vladykov 1955, was described from northeastern Italy. Its distribution is thought to include left tributaries of the River Po and the river basins of the Adriatic Sea from the River Po to the River Isonzo/Soča in Italy, Switzerland and Slovenia. It also shows a geographically isolated distribution in the Potenza River on the Adriatic slope in Central Italy. Lampetra from the Neretva River system in Croatia and Bosnia and Herzegovina and the Morača River system in Montenegro that were previously identified as L. zanandreai were recently described as a new species Lampetra soljani Tutman, Freyhof, Dulčić, Glamuzina & Geiger 2017 based on morphological data and a genetic distance between the two species of roughly 2.5% in the DNA barcoding gene cytochrome oxidase I (COI). Since DNA barcodes for L. zanandreai are only available for one population from the upper Po River in northwestern Italy, we generated additional COI nucleotide sequence data of this species from Switzerland, northeastern and central Italy comprising near topotypic material and obtained GenBank sequences of the species from Slovenia to better assess the evolutionary history of the two brook lamprey species in the river basins of the Adriatic Sea. Our data show a low sequence divergence of <1% between L. zanandreai from Switzerland, northeastern and central Italy and Slovenia and the Balkan species L. soljani. However, members of the population previously identified as 'L. zanandreai' from northwest Italy are genetically highly divergent from those of L. zanandreai and likely belong to an undescribed species, L. sp. 'upper Po'. The presence of a unique and highly divergent brook lamprey lineage in the upper Po River suggests that L. zanandreai and Lampetra sp. 'upper Po' may have evolved in separate paleo drainages during the formation of the modern Po Valley subsequent to marine inundations in the Pliocene.

Sperm competition experiments reveal low prezygotic postmating isolation between parasitic and nonparasitic lamprey ecotypes.

The role of postmating sexual selection as a potential reproductive barrier in speciation is not well understood. Here, we studied the effects of sperm competition and cryptic female choice as putative postmating barriers in two lamprey ecotypes with a partial reproductive isolation. The European river lamprey Lampetra fluviatilis is anadromous and parasitic of other fish species, whereas the brook lamprey Lampetra planeri is freshwater resident and nonparasitic. We measured sperm traits in both ecotypes and designed sperm competition experiments to test the occurrence of cryptic female choice. We also performed sperm competition experiments either at equal semen volume or equal sperm number to investigate the role of sperm velocity on fertilization success. We observed distinct sperm traits between ecotypes with a higher sperm concentration and a lower sperm velocity for L. planeri compared with L. fluviatilis. The outcomes of sperm competition reflected these differences in sperm traits, and there was no evidence for cryptic female choice irrespective of female ecotype. At equal semen volume, L. planeri males had a higher fertilization success than L. fluviatilis and vice versa at equal sperm number. Our results demonstrate that different sperm traits between ecotypes can influence the male reproductive success and thus gene flow between L. planeri and L. fluviatilis. However, postmating prezygotic barriers are absent and thus cannot explain the partial reproductive isolation between ecotypes.

A SNP marker to discriminate the european brook lamprey (Lampetra planeri), river lamprey (L. fluviatilis) and their hybrids.

The European River lamprey Lampetra fluviatilis and the brook lamprey L. planeri are two closely related species that are also considered as partially reproductively isolated ecotypes. At the larval stage, they cannot be distinguished morphologically or genetically by mitochondrial DNA. We aimed at developing a molecular tool to identify early life stages of L. fluviatilis and L. planeri. We first identified Single Nucleotide Polymorphism (SNP) markers with distinct alleles between L. fluviatilis and L. planeri based on RADseq data from 186 individuals collected in France. Then, we developed a quantitative PCR protocol to genotype a species-specific SNP, which was tested on 270 samples including larvae and adults from both species and 17 sites from Western Europe. Results were consistent with morphological identification in all cases except for samples from the Rhône drainage and the Loch Lomond in Scotland. This marker allows the identification of larval stages of L. fluviatilis, L. planeri as well as hybrids. The two cases where the marker was not diagnostic might be explained by an isolated and distinct L. planeri lineage in the Rhône drainage, and high levels of admixture among L. fluviatilis, L. planeri and the lake-parasitic form in Loch Lomond.

Биология рыб среднего течения реки Киренги

Kirenga River is the large (746 km long) right tributary of upper part of Lena River. River headwaters situated relatively close to west coast of Lake Baikal among the high peaks of the Baikal Ridge at 1470 m a.s.l. Fishes were collected from the Kirenga’s middle reaches during the investigations of biota of Tukolon’ nature protected territory along the 25 km section of riverbed (starting from approx. 375 km upstream from the mouth) in June 2013, June and September 2014, Oktober 2015 and April 2016. According to available data the ichthyofauna at this river section consists of 20 species in 11 families: Far Eastern brook lamprey Lethenteron reissneri, dace Leuciscus leuciscus baicalensis, roach Rutilus rutilus, Czekanowski's minnow Rhynchocypris czekanowskii, Amur minnow Rhynchocypris lagowskii, Eurasian minnow Phoxinus phoxinus, loach Cobitis melanoleuca, Siberian stone loach Barbatula toni, Northern pike Esox luceus, whitefish Coregonus pidschian, tugun Coregonus tugun, roundfish Prosopium cylindraceum, Baikal-Lena grayling Thymallus baicalolenensis, taimen Hucho taimen, lenok Brachymystax lenok, burbot Lota lota, sculpin (spotted sculpin, alpine bullhead) Cottus cf. poecilopus, Siberian sculpin Cottus sibiricus, ruff Gymnocephalus cernuus, perch Perca fluviatilis. Lamprey, burbot and sculpin in our findings were reported from guts of other piscivorous fich species but tugun and Czekanowski's minnow were not registered. Lenok and Baikal-Lena grayling are still most numerous in the river. Low abundance is characteristic to lamprey, loach, Siberian stone loach, burbot, sculpin and Siberian sculpin. Recent growth features in most species are generally similar with the ones for fishes from Kirenga’s low reaches and main course of Upper Lena. Their ages of maturation mostly coincides with other large streams in East Siberia.

Immunohistochemical Characterization of the Development of Long Photoreceptor Cells in the Lamprey Retina.

The adult lamprey retina has two types of photoreceptor cells, short and long photoreceptor cells, which are equivalent to rods and cones of other vertebrates. In contrast, the retina of lamprey larvae only contains a single type of photoreceptor cell, which appears to correspond to the short photoreceptor cell. However, the developmental pattern of the long photoreceptor cell is unknown. Previously, we reported that antibodies against rhodopsin and iodopsin (the chicken red cone opsin) could discriminate between the outer segments of short and long photoreceptor cells, respectively, in the retina of adult Japanese river lamprey (Lethenteron camtschaticum). Here, we immunohistochemically investigate the appearance of long photoreceptor cells in the larval and adult retinas of the Far Eastern brook lamprey (Lethenteron reissneri), which is a close relative of the Japanese river lamprey, by using anti-iodopsin antibody. We found that iodopsin immunoreactivity was localized not only in the adult retina but also in the larval retina. Moreover, we examined the immunohistochemical localization of signal transduction molecules, such as transducin and arrestin, in the iodopsin-immunoreactive cells of the larval retina. The iodopsin-immunoreactive cells also contained both transducin and arrestin, suggesting that long photoreceptor cells are already functional in the larval stage before the acquisition of visual function. Our results suggest that the iodopsin-immunoreactive cells may be related to not only cone vision in the adult but also photoreception in the larval lamprey.

Transcriptomic analysis of gonadal development in parasitic and non-parasitic lampreys (Ichthyomyzon spp.), with a comparison of genomic resources in these non-model species.

Lampreys are jawless fishes that diverged ∼550 million years ago from other vertebrates. Sequencing of the somatic and the germline genomes of the sea lamprey (Petromyzon marinus) in 2013 and 2018, respectively, has helped to improve our understanding of the genes and gene networks that control many aspects of lamprey development. However, little is known about the genetic basis of gonadal differentiation in lampreys, partly due to the prolonged period during which their gonads remain sexually indeterminate. We performed RNA-sequencing on gonadal samples from four chestnut lamprey (Ichthyomyzon castaneus) and six northern brook lamprey (I. fossor) to identify differentially expressed genes (DEG’s) and pathways associated with transcriptomic differences in: (1) larvae during early gonadal differentiation versus definitive females (i.e., with oocytes in the slow cytoplasmic growth phase); and (2) females versus definitive males undergoing spermatogonial proliferation. We compared the mapping percentages of these transcriptomes to the two available sea lamprey reference genomes and three annotation files (Ensembl and UCSC for the somatic genome and SIMRbase for the germline genome). We found that mapping the RNA-seq reads to the germline genome gave superior results and, using Trinotate, we provided new putative annotations for 8161 genes in the somatic assembly and 880 genes for the germline assembly. We identified >2000 DEG’s between stages and sexes, as well as biological pathways associated with each. Interestingly, some of the upregulated genes (e.g., DEG’s associated with spermiation) suggest that changes in gene expression can precede morphological changes by several months. In contrast, only 81 DEG’s were evident between the chestnut lamprey (that remains sexually immature during an extended post-metamorphic parasitic feeding phase) and the nonparasitic northern brook lamprey (that undergoes sexual maturation near the end of metamorphosis), but few replicates were available for comparable stages and sexes. This work lays the foundation for identifying and confirming the orthology and the function of genes involved in gonadal development in these and other lamprey species across more developmental stages.

Changes in native lamprey populations in the Great Lakes since the onset of sea lamprey (Petromyzon marinus) control

The control of invasive sea lamprey in the Great Lakes basin has been highly successful, but has deleteriously affected native lamprey species. American brook, northern brook, silver and chestnut lampreys are all susceptible to lampricide treatments. Incidental catch from surveys that targeted sea lamprey were used to determine changes in distributions and catch rates of these native species within the Great Lakes basin since the onset of the sea lamprey control program. Over 2.4 million native lampreys were collected during sea lamprey control operations between 1947 and 2018. Native lampreys were no longer captured from many treated and untreated streams. Silver lamprey and northern brook lamprey distributions were reduced most substantially. These species were no longer found in about three quarters of the number of streams in which they were historically present. The number of streams inhabited by chestnut lamprey has diminished by half. American brook lamprey were still present in the majority of streams previously occupied. Catch rates declined in northern brook lamprey and Ichthyomyzon larvae in some Great Lakes basins, but changes in collection gear and methods made comparisons difficult. Trap and weir data show drastic declines in silver lamprey and chestnut lamprey. Streams that have not been treated with lampricide have suffered similar or greater declines in both the number of streams occupied and catch rates for all four species when compared to untreated streams. Therefore reductions in native lamprey populations were likely due to a number of environmental changes, and not exclusively from lampricide treatments.

Riverscape genetics in brook lamprey: genetic diversity is less influenced by river fragmentation than by gene flow with the anadromous ecotype.

Understanding the effect of human-induced landscape fragmentation on gene flow and evolutionary potential of wild populations has become a major concern. Here, we investigated the effect of riverscape fragmentation on patterns of genetic diversity in the freshwater resident European brook lamprey (Lampetra planeri) that has a low ability to pass obstacles to migration. We tested the hypotheses of (i) asymmetric gene flow following water current and (ii) an effect of gene flow with the closely related anadromous river lamprey (L. fluviatilis) ecotype on L. planeri genetic diversity. We genotyped 2472 individuals, including 225 L. fluviatilis, sampled from 81 sites upstream and downstream barriers to migration, in 29 western European rivers. Linear modelling revealed a strong positive relationship between genetic diversity and the distance from the river source, consistent with expected patterns of decreased gene flow into upstream populations. However, the presence of anthropogenic barriers had a moderate effect on spatial genetic structure. Accordingly, we found evidence for downstream-directed gene flow, supporting the hypothesis that barriers do not limit dispersal mediated by water flow. Downstream L. planeri populations in sympatry with L. fluviatilis displayed consistently higher genetic diversity. We conclude that genetic drift and slight downstream gene flow drive the genetic make-up of upstream L. planeri populations whereas gene flow between ecotypes maintains higher levels of genetic diversity in L. planeri populations sympatric with L. fluviatilis. We discuss the implications of these results for the design of conservation strategies of lamprey, and other freshwater organisms with several ecotypes, in fragmented dendritic river networks.

Infection of the Carpathian brook lamprey (Eudontomyzon danfordi Regan, 1911) with a dermocystid parasite in the Tisza River Basin, Hungary.

The Carpathian brook lamprey (Eudontomyzon danfordi Regan, 1911) is an endemic protected species of Cephalaspidomorphi in the Carpathian Basin. No parasites have become known from these jawless vertebrates to date. Here, the authors describe an infection from a single specimen manifesting in protuberant skin cysts 7-10mm in diameter, scattered on the body surface. Similar dermal infection was observed in 25 of the 274 lampreys recorded in the population survey. Skin cysts filled with round spore-like structures of a dermocystid parasite were found. These particles measured 8-14µm in diameter and had an about 0.5µm thick wall, and containing mainly a granular mass and a relatively scarce plasma. No hyphae were recorded. Despite conspicuous morphological changes in the skin, no inflammatory reactions were found. The molecular analysis of 18S rDNA showed similarity to dermocystid species of several fish species but differed from them approximately by 2%. This is the first record of a dermocystid parasite infecting a jawless vertebrate.

The non-visual opsins expressed in deep brain neurons projecting to the retina in lampreys

In lower vertebrates, brain photoreceptor cells express vertebrate-specific non-visual opsins. We previously revealed that a pineal-related organ-specific opsin, parapinopsin, is UV-sensitive and allows pineal wavelength discrimination in lampreys and teleost. The Australian pouched lamprey was recently reported as having two parapinopsin-related genes. We demonstrate that a parapinopsin-like opsin from the Japanese river lamprey exhibits different molecular properties and distribution than parapinopsin. This opsin activates Gi-type G protein in a mammalian cell culture assay in a light-dependent manner. Heterologous action spectroscopy revealed that the opsin forms a violet to blue-sensitive pigment. Interestingly, the opsin is co-localised with green-sensitive P-opsin in the cells of the M5 nucleus of Schober (M5NS) in the mesencephalon of the river and brook lamprey. Some opsins-containing cells of the river lamprey have cilia and others an axon projecting to the retina. The opsins of the brook lamprey are co-localised in the cilia of centrifugal neurons projecting to the retina, suggesting that cells expressing the parapinopsin-like opsin and P-opsin are sensitive to violet to green light. Moreover, we found neural connections between M5NS cells expressing the opsins and the retina. These findings suggest that the retinal activity might be modulated by brain photoreception.

Reference databases, primer choice, and assay sensitivity for environmental metabarcoding: Lessons learnt from a re‐evaluation of an eDNA fish assessment in the Volga headwaters

AbstractBiodiversity monitoring via environmental DNA, particularly metabarcoding, is evolving into a powerful assessment tool for riverine systems. However, for metabarcoding to be fully integrated into standardized monitoring programmes, some current challenges concerning sampling design, laboratory workflow, and data analysis need to be overcome. Here, we review some of these major challenges and potential solutions. We further illustrate three potential pitfalls, namely the choice of suitable metabarcoding primers, the necessity of complete reference databases, and varying assay sensitivities, by a reappraisal of our‐own recently carried out metabarcoding study in the Volga headwaters. TaqMan qPCRs had detected catfish (Silurus glanis) and European eel (Anguilla anguilla), whereas metabarcoding had not, in the same samples. Furthermore, after extending the genetic reference database by 12 additional species and re‐analysing the metabarcoding data, we additionally detected the Siberian spiny loach (Cobitis sibirica) and Ukrainian brook lamprey (Eudontomyzon mariae) and reassigned the operational taxonomic units previously assigned to Misgurnus fossilis to Cobitis sibirica. In silico analysis of metabarcoding primer efficiencies revealed considerable variability among primer pairs and among target species, which could lead to strong primer bias and potential false‐negatives in metabarcoding studies if not properly compensated for. These results highlight some of the pitfalls of eDNA‐metabarcoding as a means of monitoring fish biodiversity in large rivers, which need to be considered in order to fully unleash the full potential of these approaches for freshwater biodiversity monitoring.

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.

An experimental study to evaluate predation threats on two native larval lampreys in the Columbia River Basin, USA

AbstractExperimental predation studies were conducted to evaluate and compare the predation threats of 10 species of native and non‐native fishes on larvae of Pacific Lamprey, Entosphenus tridentatus, and Western Brook Lamprey, Lampetra richardsoni. The relative predatory threats were examined over four sets of binary factors, including the following: (a) short (2‐day) or long (7‐day) duration, (b) presence/absence of fine sediment, (c) live or dead larvae and (d) species of lampreys. Our short‐term results showed a positive correlation with the sizes of predator fishes and consumed lamprey larvae. Also, most predator fishes had a significantly higher propensity to prey on lamprey larvae when sediment was absent. Conversely, this demonstrated the importance of sediment in protecting lamprey larvae from predation. Based on the predatory behaviour for live and dead larvae, predator fishes were classified into four groups using principal component analysis. Predation rates of larvae in sediment by piscivorous predators including Northern Pikeminnow Ptychocheilus oregonensis and Smallmouth Bass Micropterus dolomieu were lower and showed no differences even when the study duration was extended. In contrast, predation rates by benthic predators including White Sturgeon Acipenser transmontanus and Common Carp Cyprinus carpio, increased when we extended the study duration. This suggests that when given longer duration, benthic predators are more adept at consuming larvae within the sediment. These results provide important context for assessing the potential threat of predation on lampreys in streams, which is a key knowledge gap for lampreys.

Novel “omega muscle units” in superficial body-wall myotomes during metamorphosis in the northern brook lamprey (Ichthyomyzonfossor)

Lampreys transform from sedentary filter feeders to more mobile adults through a dramatic metamorphosis that includes remodeling of head muscle and skeletal systems. Metamorphic modifications of body-wall myotomes that could support changes in swimming behavior from larvae to adults have not been previously reported. Thus, transverse sections of northern brook lamprey (Ichthyomyzon fossor Reighard and Cummins, 1916) in larval (n = 4), metamorphosing (n = 3), and adult (n = 2) stages were used to investigate the architecture of body-wall muscle and to detect whether Pax7 and MyoD, proteins important in myogenesis, were co-localized in any muscle nuclei. In addition to myotomal complexity of muscle units composed of parietal and central fibers, there was a novel pattern of omega-shaped muscle units with curves of muscle fibers in the superficial mid-body myotome in metamorphosing lamprey. Small satellite-like cells were identified on central fibers in metamorphosing and adult lamprey muscle using routine histology and immunolocalization of Pax7 and MyoD with antibodies that specifically detect mammalian and teleost proteins. Transient “omega muscle units” may be a marker for impending myotomal growth and increasing swimming efficiency during maturation, possibly restricted to metamorphosis. Finding satellite-like cells suggests that Pax7 and MyoD may have distinctive roles in lamprey myogenesis.

Multiple stable isotopes identify sediment organic matter as the primary nutritional source to American brook lamprey larvae along a stream gradient

Multiple stable isotope ratios (δ2H, δ13C, δ15N, and δ34S) of American brook lamprey (Lethenteron appendix) ammocoetes and their potential sources were measured along a stream gradient. Isotopic ratios were successful at differentiating three potential sources: terrestrial organic matter (TOM; upland plants), sediment organic matter (SOM; benthic organic material), and algae. Bayesian mixing models (MixSIAR) utilizing δ13C, δ15N, and δ34S estimated ammocoetes were highly reliant on SOM at all sites (&gt;95%), but inclusion of δ2H increased the importance of both algae and model uncertainty. Ammocoete δ2H was more depleted than anticipated even after lipid extraction and after accounting for trophic water contributions, suggesting δ2H in consumer tissues (especially for long-lived animals) is still incompletely understood. Model sensitivity testing with trophic fractionation values did not substantially change the results. Regardless of the mixing model investigated, none indicated more algae in ammocoete diets in downstream rather than upstream sites, in contrast with expectations about river productivity. Stable isotopes suggest SOM, and not algae, is nutritionally most important to ammocoetes, even in downstream reaches of rivers.

Brook lamprey survival in the dry riverbed of an intermittent stream

Intermittent streams account for approximately one half of the global river network, and are especially common in arid and semi-arid regions. These systems are inhabited by several fish species that are adapted to their natural flow intermittency. While many fish species rely on permanent pools or perennial reaches to survive during the dry season, our knowledge regarding fish survival strategies in dry reaches is incomplete. Our observations revealed that ammocoetes (i.e. lamprey larvae) of a non-parasitic brook lamprey (Lampetra sp.) survived in the hyporheic zone of an intermittent stream in the absence of surface water for at least 22 days. This is the first reported observation of ammocoetes persisting in the hyporheic water after loss of surface water, a behavior that has been previously reported for just a few fish species.

Stream characteristics associated with feeding type in silver (Ichthyomyzon unicuspis) and northern brook (I. fossor) lampreys and tests for phenotypic plasticity

In most lamprey genera, “paired” species exist in which the larvae are morphologically indistinguishable but adult feeding type differs. The lack of diagnostic genetic differences in many pairs has led to suggestions that they constitute a single gene pool with environmentally influenced feeding types. To investigate whether stream characteristics are correlated with feeding type in the parasitic silver lamprey Ichthyomyzon unicuspis and non-parasitic northern brook lamprey I. fossor, eight variables (pH, alkalinity, conductivity, discharge, total dissolved solids, and density of larval sea lamprey Petromyzon marinus, Ichthyomyzon spp., and total larval density) were compared among eight streams with only silver lamprey, 10 with only northern brook lamprey, and 13 with both species, using classification tree analysis. The most parsimonious model had a 24% misclassification rate, with silver lamprey tending to inhabit streams with higher sea lamprey larval density and northern brook lamprey tending to inhabit streams with higher Ichthyomyzon larval density. We then conducted a pilot study investigating phenotypic plasticity in a lab-based common garden experiment and an in situ transplant experiment. These studies encountered myriad difficulties and were unable to demonstrate plasticity, but did identify challenges associated with culturing Ichthyomyzon larvae. Development of effective rearing procedures for Ichthyomyzon lampreys is essential for any future similar studies.