Identification of long-preserved specimens reveals the historical geographic range of the Patagonian lamprey Geotria macrostoma (Burmeister, 1868) in southern South America.

The lamprey genus Geotria Gray, 1851 currently includes only two species: G. australis and G. macrostoma. However, taxonomic relationships within the genus have traditionally been ambiguous and difficult to establish due to the extreme changes in morphology, dentition, and coloration that lampreys undergo during their life cycles, particularly during upstream migration and sexual maturation. Consequently, several lamprey specimens held in museum collections have remained unidentified, especially those from Argentina. In this study, a series of morphometric characters were subjected to discriminant function analysis (DFA) to identify the lamprey species collected during 1867-2004 from the de la Plata River and Patagonia. These specimens are housed at the Museo Argentino de Ciencias Naturales "Bernardino Rivadavia" in Buenos Aires, the Museo de Historia Natural de Montevideo, and the Naturhistoriska riksmuseet in Stockholm. Based on the proportions of the length of the oral disc, prebranchial, and pre-caudal body regions, and the depth of the trunk, DFA provided conclusive evidence that the specimens corresponded to the recently revalidated G. macrostoma (Burmeister, 1868), which was originally incorrectly named as Petromyzon macrostomus Burmeister, 1868, Exomegas macrostomus (Berg, 1899), Geotria chilensis (Berg, 1895), and Geotria macrostoma f. gallegensis Smitt, 1901, as well as other nontype museum individuals of uncertain taxonomic status. The identifications of these long-preserved museum specimens provided key information on the historical geographic range of Argentinian lampreys and suggest that the disappearance of the species reported from northern localities (the Pampean Region) can be attributed to the degradation of their critical habitats, primarily caused by anthropogenic impact and climate change.

The effects of biochar aging on rhizosphere microbial communities in cadmium-contaminated acid soil.

Biochars are widely used in the remediation of Cd-contaminated soils. However, changes in the bacterial communities in the rhizosphere contaminated with Cd in response to biochar aging are poorly studied. Addressing this gap in knowledge is important to improving micro-ecological services on healthy growth of plants with mitigation strategies against Cd contamination. An aging experiment (270 days) was conducted with biochars derived from poultry litter and sugar-gum wood added to a Cd-contaminated acid soil. Bacterial communities in the rhizosphere of Brassica rapa and bulk soils were investigated after 1, 90 and 270 days of biochar aging. There was no significant difference (P > 0.05) in bacterial Shannon and Simpson indices between the control and biochar treatments. However, compared to the no-Cd control, the addition of Cd decreased the relative abundances of Firmicutes, Chloroflexi and Acidobacteriota but increased those of Actinobacteriota and Proteobacteria. Poultry-litter biochar had the largest effect on bacterial community composition, especially in the rhizosphere. Aging of poultry-litter biochar increased the abundance of Armatimonadota over time more than the sugar-gum-wood biochar, which was attributed to a lower pH and higher bioavailability of Cd in the sugar-gum-wood biochar treatment. The addition of poultry-litter biochar to the contaminated soil mitigated the bioaccumulation of Cd by increasing soil pH and restoring soil bacterial ecology in contaminated acid soils over time.

On the invalid resurrection of the lamprey genus Exomegas Gill, 1883.

The lamprey genus Exomegas Gill, 1883, was erected on the assumption that it was distinguishable from Geotria Gray, 1851, by possessing three rather than two cusps on the transverse lingual lamina (TLL). Based on literature review and examination of holotypes and new data, the authors reaffirm that the TLL of Geotria possesses two or three cusps in the adult stage. The reduction or disappearance of the middle cusp at the beginning or during the spawning run constitutes a key feature of Geotria. The resurrection of Exomegas by Firpo Lacoste, Fernández and Scioscia, Journal of Fish Biology, 2021, 1-6, 1507-1512, is therefore unjustified and not supported.

Morphometric and physical characteristics distinguishing adult Patagonian lamprey, Geotria macrostoma from the pouched lamprey, Geotria australis.

The pouched lamprey, Geotria australis Gray, 1851, has long been considered monotypic in the Geotriidae family with a wide southern temperate distribution across Australasia and South America. Recent studies have provided molecular and morphological evidence for a second Geotria species in South America; Geotria macrostoma (Burmeister, 1868). The aim of this study was to determine morphometric and physical characteristics of adult G. macrostoma that further differentiate this re-instated species of Geotriidae from G. australis. The diagnostic features discriminating immature adult G. macrostoma from G. australis when entering fresh water, are distinct differences in dentition, oral papillae and fimbriae counts and differences in coloration. In addition, G. macrostoma display greater growth of the prebranchial region and oral disc and has a deeper body depth and higher condition factor. All current ecological knowledge of the genus Geotria is based on Australasian populations, which may not be applicable to G. macrostoma. To ensure the conservation and protection of the Patagonian lamprey as a re-identified species, further investigations are needed to understand its life history, biology and ecology throughout its range.

Large variations in eutrophication among estuaries reflect massive differences in composition and biomass of macroalgal drift.

The characteristics of detached macroalgae (drift) in nearby highly eutrophic and mesotrophic estuaries in south-western Australia are compared to elucidate the magnitude and types of changes that occur in macroalgal drift when estuaries receive excessive nutrient input. Drift characteristics in the large basins of the microtidal, eutrophic Peel-Harvey and mesotrophic Swan-Canning, which is not subjected to large nutrient inputs directly from agricultural land, differed markedly. Biomass (dry weight) in mesotrophic estuary was dominated by rhodophytes (92%), particularly Laurencia and Hypnea, and in eutrophic estuary by opportunistic chlorophytes (68%), especially Chaetomorpha and Ulva. Prevalence and biomass of drift were far greater in the eutrophic estuary, particularly during summer and autumn when macroalgal growth rose sharply. Macroalgal biomass in the eutrophic estuary was positively related to salinity. These results facilitate predictions of how climatic and other anthropogenic changes influence extent of macroalgal growth and thus change the estuarine environment.

Visual opsin expression and morphological characterization of retinal photoreceptors in the pouched lamprey (Geotria australis, Gray).

Lampreys are extant members of the agnathan (jawless) vertebrates that diverged ~500 million years ago, during a critical stage of vertebrate evolution when image-forming eyes first emerged. Among lamprey species assessed thus far, the retina of the southern hemisphere pouched lamprey, Geotria australis, is unique, in that it possesses morphologically distinct photoreceptors and expresses five visual photopigments. This study focused on determining the number of different photoreceptors present in the retina of G. australis and whether each cell type expresses a single opsin class. Five photoreceptor subtypes were identified based on ultrastructure and differential expression of one of each of the five different visual opsin classes (lws, sws1, sws2, rh1, and rh2) known to be expressed in the retina. This suggests, therefore, that the retina of G. australis possesses five spectrally and morphologically distinct photoreceptors, with the potential for complex color vision. Each photoreceptor subtype was shown to have a specific spatial distribution in the retina, which is potentially associated with changes in spectral radiance across different lines of sight. These results suggest that there have been strong selection pressures for G. australis to maintain broad spectral sensitivity for the brightly lit surface waters that this species inhabits during its marine phase. These findings provide important insights into the functional anatomy of the early vertebrate retina and the selection pressures that may have led to the evolution of complex color vision.

Mechanisms for the removal of Cd(II) and Cu(II) from aqueous solution and mine water by biochars derived from agricultural wastes.

The capacity of biochars derived from agricultural wastes to remove Cd(II) and Cu(II) from aqueous solution and contaminated mine water was evaluated using laboratory-based batch sorption experiments. To examine immobilization of heavy metals, biochars produced in a commercial-scale mobile pyrolizer from feedstocks: poultry litter; lucerne shoot; vetch shoot; canola shoot; wheat straws; and sugar-gum wood, were tested in a liquid-based system. Biochars were characterized by FTIR, XPS and XRD before and after the mine water treatment. Lucerne biochar had the highest Langmuir sorption capacity of Cd(II) (6.28 mg g-1) and vetch-derived biochar had the highest Cu(II) sorption capacity (18.0 mg g-1) at pH 5.5. All the biochars exhibited higher sorption capacity for Cu(II) than for Cd(II). The smaller ionic radius and higher electronegativity of Cu(II), and the PO43-, CO32- and N-containing functional groups of biochars enhanced their binding affinity. The results demonstrated that poultry litter-derived biochar was effective at removal of the Cd(II) and Cu(II) from mine water up to the levels recommended by the World Health Organisation. The results revealed that precipitation with CO32- and PO43-, complexation with -OH and -COOH groups and electrostatic interaction with O-containing surface functional groups were the main mechanisms involved in the removal of multi-metals by biochars, and that selection of feedstock materials for biochar production is important to maximise remediation of multi-metals in contaminated water.

Lipid and protein catabolism contribute to aerobic metabolic responses to exhaustive exercise during the protracted spawning run of the lamprey Geotria australis.

This paper has integrated new and past data to elucidate how lipid, protein and glycogen metabolism contribute to generating the ATP required by the southern hemisphere lamprey Geotria australis during its ~ 13-15 months, non-trophic upstream spawning migration. Energy is required for maintenance, swimming, the development of gonads and secondary sexual characters and spawning and post-spawning activities. Plasma and muscle metabolites were measured in animals subjected to an exercise-recovery regime at the commencement and completion of the spawning run. The present study demonstrated the following. At all stages of the migration, plasma glucose and glycerol concentrations increased during exercise and then declined, whereas plasma FFAs exhibited the reverse trend. During exercise and recovery, alanine declined and ammonia increased in the plasma of early migrants, while the opposite occurred in mature males. Following exercise, muscle alanine rose and then declined in early migrants, but declined and then rose in mature males. The composite data emphasise that, while the same catabolic processes are employed by both sexes early in the migration, when animals are immature, they differ markedly between the sexes as they mature and then spawn, reflecting their different demands. Energy is supplied predominantly via anaerobic metabolism in early migrants, but by anaerobic and aerobic metabolism in prespawning females and by aerobic metabolism in mature males and spent females. Although proteolysis is limited early in the migration, it is employed extensively during maturation and particularly by females, which undergo a substantial reduction in length in the lead-up to spawning.

Zooplankton dynamics in a highly eutrophic microtidal estuary.

Mesozooplankton was sampled seasonally in a large microtidal estuary (Peel-Harvey) suffering from massive macroalgal growths and cyanobacterial blooms. Comparisons with other estuaries indicate that eutrophication led to copepod abundance declining and macroalgal-associated species increasing. Mesozooplankton species are almost exclusively autochthonous, i.e. spend entire life cycle within the estuary. Meroplanktonic species are virtually absent because main benthic macroinvertebrate species undergo direct benthic rather than planktotrophic development. There are also few abundant holoplanktonic species. Most species are tychoplanktonic, i.e. benthic and transported into plankton through physical disturbance of sediment. Species number, concentration and Simpson's Index are greater during night than day. Annual cyclical changes in species composition are related closely to changes in salinity. At the most degraded site, nematode concentrations were high and the species number and concentration changed markedly during extreme eutrophication, when oxygen concentrations were low, disrupting annual cyclical changes in species composition.

Microtidal estuaries warrant special management measures that recognise their critical vulnerability to pollution and climate change.

Not all estuaries are equally susceptible to anthropogenic perturbation. Microtidal estuaries with long residence times are intrinsically less robust than well-flushed macrotidal estuaries, facilitating the accumulation of contaminants. This promotes development of blooms of non-toxic and toxic phytoplankton, and hypoxia and anoxia may occur in deeper sections of the typically stratified water column. In Mediterranean and arid climates, high temperatures and low and/or seasonal rainfall can result in marked hypersalinity. Thus, any increase in anthropogenic perturbation will further decrease the health of a system in which the biota already experiences natural stress. Microtidal estuaries are also more susceptible to climate change, the detrimental longer-term effects of which are becoming manifestly obvious. Numerous attempts have been made to develop novel solutions to problems caused by eutrophication, phytoplankton blooms, hypoxia and hypersalinity, which have met with various levels of success, but the need for such measures and effective legislation is increasingly critical.

Growth, condition, and maturity schedules of an estuarine fish species change in estuaries following increased hypoxia due to climate change.

Understanding challenges posed by climate change to estuaries and their faunas remains a high priority for managing these systems and their communities. Freshwater discharge into a range of estuary types in south-western Australia between 1990 and 2015 is shown to be related to rainfall. This largely accounts for decreases in discharge in this microtidal region being more pronounced on the west coast than south coast, where rainfall decline was less. Results of an oxygen-balance model imply that, as demonstrated by empirical data for the Swan River Estuary, declines in discharge into a range of estuary types would be accompanied by increases in the extent of hypoxia. In 2013-15, growth and body condition of the teleost Acanthopagrus butcheri varied markedly among three permanently open, one intermittently-open, one seasonally-closed and one normally-closed estuary, with average time taken by females to reach the minimum legal length (MLL) of 250 mm ranging from 3.6 to 17.7 years. It is proposed that, in a given restricted period, these inter-estuary variations in biological characteristics are related more to differences in factors, such as food resources and density, than to temperature and salinity. The biological characteristics of A. butcheri in the four estuaries, for which there are historical data, changed markedly between 1993-96 and 2013-15. Growth of both sexes, and also body condition in all but the normally-closed estuary, declined, with females taking between 1.7 and 2.9 times longer to attain the MLL. Irrespective of period, body condition, and growth are positively related. Age at maturity typically increased between periods, but length at maturity declined only in the estuary in which growth was greatest. The plasticity of the biological characteristics of A. butcheri, allied with confinement to its natal estuary and ability to tolerate a wide range of environmental conditions, makes this sparid and comparable species excellent subjects for assessing estuarine "health."

The Central Nervous System of Jawless Vertebrates: Encephalization in Lampreys and Hagfishes.

Lampreys and hagfishes are the sole surviving representatives of the early agnathan (jawless) stage in vertebrate evolution, which has previously been regarded as the least encephalized group of all vertebrates. Very little is known, however, about the extent of interspecific variation in relative brain size in these fishes, as previous studies have focused on only a few species, even though lampreys exhibit a variety of life history traits. While some species are parasitic as adults, with varying feeding behaviors, others (nonparasitic species) do not feed after completing their macrophagous freshwater larval phase. In addition, some parasitic species remain in freshwater, while others undergo an anadromous migration. On the basis of data for postmetamorphic individuals representing approximately 40% of all lamprey species, with representatives from each of the three families, the aforementioned differences in life history traits are reflected in variations in relative brain size. Across all lampreys, brain mass increases with body mass with a scaling factor or slope (α) of 0.35, which is less than those calculated for different groups of gnathostomatous (jawed) vertebrates (α = 0.43-0.62). When parasitic and nonparasitic species are analyzed separately, with phylogeny taken into account, the scaling factors of both groups (parasitic α = 0.43, nonparasitic α = 0.45) approach those of gnathostomes. The relative brain size in fully grown adults of parasitic species is, however, less than that of the adults of nonparasitic species, paralleling differences between fully grown adults and recently metamorphosed individuals of anadromous species. The average degree of encephalization is found in anadromous parasitic lampreys and might thus represent the ancestral condition for extant lampreys. These results suggest that the degree of encephalization in lampreys varies according to both life history traits and phylogenetic relationships.

Retinal temporal resolution and contrast sensitivity in the parasitic lamprey Mordacia mordax and its non-parasitic derivative Mordacia praecox.

Lampreys and hagfishes are the sole extant representatives of the early agnathan (jawless) vertebrates. We compared retinal function of fully metamorphosed, immature Mordacia mordax (which are about to commence parasitic feeding) with those of sexually mature individuals of its non-parasitic derivative Mpraecox We focused on elucidating the retinal adaptations to dim-light environments in these nocturnally active lampreys, using electroretinography to determine the temporal resolution (flicker fusion frequency, FFF) and temporal contrast sensitivity of enucleated eyecups at different temperatures and light intensities. FFF was significantly affected by temperature and light intensity. Critical flicker fusion frequency (cFFF, the highest FFF recorded) of M. praecox and M. mordax increased from 15.1 and 21.8 Hz at 9°C to 31.1 and 36.9 Hz at 24°C, respectively. Contrast sensitivity of both species increased by an order of magnitude between 9 and 24°C, but remained comparatively constant across all light intensities. Although FFF values for Mordacia spp. are relatively low, retinal responses showed a particularly high contrast sensitivity of 625 in M. praecox and 710 in M. mordax at 24°C. This suggests selective pressures favour low temporal resolution and high contrast sensitivity in both species, thereby enhancing the capture of photons and increasing sensitivity in their light-limited environments. FFF indicated all retinal photoreceptors exhibit the same temporal response. Although the slow response kinetics (i.e. low FFF) and saturation of the response at bright light intensities characterise the photoreceptors of both species as rod-like, it is unusual for such a photoreceptor to be functional under scotopic and photopic conditions.

Ontogenetic shifts in brain scaling reflect behavioral changes in the life cycle of the pouched lamprey Geotria australis.

Very few studies have described brain scaling in vertebrates throughout ontogeny and none in lampreys, one of the two surviving groups of the early agnathan (jawless) stage in vertebrate evolution. The life cycle of anadromous parasitic lampreys comprises two divergent trophic phases, firstly filter-feeding as larvae in freshwater and secondly parasitism as adults in the sea, with the transition marked by a radical metamorphosis. We characterized the growth of the brain during the life cycle of the pouched lamprey Geotria australis, an anadromous parasitic lamprey, focusing on the scaling between brain and body during ontogeny and testing the hypothesis that the vast transitions in behavior and environment are reflected in differences in the scaling and relative size of the major brain subdivisions throughout life. The body and brain mass and the volume of six brain structures of G. australis, representing six points of the life cycle, were recorded, ranging from the early larval stage to the final stage of spawning and death. Brain mass does not increase linearly with body mass during the ontogeny of G. australis. During metamorphosis, brain mass increases markedly, even though the body mass does not increase, reflecting an overall growth of the brain, with particularly large increases in the volume of the optic tectum and other visual areas of the brain and, to a lesser extent, the olfactory bulbs. These results are consistent with the conclusions that ammocoetes rely predominantly on non-visual and chemosensory signals, while adults rely on both visual and olfactory cues.

Bovine pancreatic trypsin inhibitor is a new antifungal peptide that inhibits cellular magnesium uptake.

Antimicrobial peptides (AMPs) are promising agents for control of bacterial and fungal infections. Traditionally, AMPs were thought to act through membrane disruption but recent experiments have revealed a diversity of mechanisms. Here we describe a novel antifungal activity for bovine pancreatic trypsin inhibitor (BPTI). BPTI has several features in common with a subset of antimicrobial proteins in that it is small, cationic and stabilized by disulphide bonds. BPTI inhibits growth of Saccharomyces cerevisiae and the human pathogen Candida albicans. Screening of the yeast heterozygous essential deletion collection identified the magnesium transporter Alr1p as a potential BPTI target. BPTI treatment of wild type cells resulted in a lowering of cellular Mg(2+) levels. Populations treated with BPTI had fewer cells in S-phase of the cell cycle and a corresponding increase of cells in G(0)/G(1) and G(2) phases. The same patterns of cell cycle arrest obtained with BPTI were also obtained with the magnesium channel inhibitor hexamine(III)cobalt chloride. Analysis of the growth inhibition of C. albicans revealed that BPTI is inhibiting growth via the same mechanism in the two yeast species. Inhibition of magnesium uptake by BPTI represents a novel mechanism of action for AMPs.

Classification of retinal ganglion cells in the southern hemisphere lamprey Geotria australis (Cyclostomata).

Lampreys are one of two extant representatives of the earliest group of vertebrates, the agnathans or jawless fishes. The single species of the southern hemisphere lamprey family Geotriidae, Geotria australis, possesses the potential for pentachromatic color discrimination opposed to the mono- or dichromacy found in other lampreys. However, little is known of the retinal ganglion cell types that contribute to visual processing in G. australis. A quantitative morphological approach was used to distinguish and describe retinal ganglion cell types in G. australis. The morphology of retinal ganglion cells was revealed by retrograde biocytin labeling from the optic disc. Cells were digitally reconstructed, and somatic area and position and dendritic field size, density, tortuosity, and stratification were subjected to quantitative morphometric analyses. Cluster analysis, in conjunction with similarity profile analysis (SIMPROF), statistically identified five discrete monostratified retinal ganglion cell types, one of which may comprise two subtypes. Two bistratified types were identified separately, including a biplexiform and a bistratified subtype. The use of cluster analysis with SIMPROF provided a robust statistical technique for objectively identifying cell types whose characteristics were similar and significantly different from those of other types and thus provides an objective resolution of the problems posed by "lumpers vs. splitters" when designating cell types. The diversity of retinal ganglion cells suggests that visual information in the lamprey G. australis is processed in parallel streams, as in gnathostomes. These findings, together with the results of previous studies, indicate that the visual system of the lamprey G. australis represents the upper limit of visual complexity in extant agnathans.

Retinal amino acid neurochemistry of the southern hemisphere lamprey, Geotria australis.

Lampreys are one of the two surviving groups of the agnathan (jawless) stages in vertebrate evolution and are thus ideal candidates for elucidating the evolution of visual systems. This study investigated the retinal amino acid neurochemistry of the southern hemisphere lamprey Geotria australis during the downstream migration of the young, recently-metamorphosed juveniles to the sea and during the upstream migration of the fully-grown and sexually-maturing adults to their spawning areas. Glutamate and taurine were distributed throughout the retina, whilst GABA and glycine were confined to neurons of the inner retina matching patterns seen in most other vertebrates. Glutamine and aspartate immunoreactivity was closely matched to Müller cell morphology. Between the migratory phases, few differences were observed in the distribution of major neurotransmitters i.e. glutamate, GABA and glycine, but changes in amino acids associated with retinal metabolism i.e. glutamine and aspartate, were evident. Taurine immunoreactivity was mostly conserved between migrant stages, consistent with its role in primary cell functions such as osmoregulation. Further investigation of glutamate signalling using the probe agmatine (AGB) to map cation channel permeability revealed entry of AGB into photoreceptors and horizontal cells followed by accumulation in inner retinal neurons. Similarities in AGB profiles between upstream and downstream migrant of G. australis confirmed the conservation of glutamate neurotransmission. Finally, calcium binding proteins, calbindin and calretinin were localized to the inner retina whilst recoverin was localized to photoreceptors. Overall, conservation of major amino acid neurotransmitters and calcium-associated proteins in the lamprey retina confirms these elements as essential features of the vertebrate visual system. On the other hand, metabolic elements of the retina such as neurotransmitter precursor amino acids and Müller cells are more sensitive to environmental changes associated with migration.

Advanced signal processing analysis of laser-induced breakdown spectroscopy data for the discrimination of obsidian sources.

Obsidian is a natural glass of volcanic origin and a primary resource used by indigenous peoples across North America for making tools. Geochemical studies of obsidian enhance understanding of artifact production and procurement and remain a priority activity within the archaeological community. Laser-induced breakdown spectroscopy (LIBS) is an analytical technique being examined as a means for identifying obsidian from different sources on the basis of its 'geochemical fingerprint'. This study tested whether two major California obsidian centers could be distinguished from other obsidian localities and the extent to which subsources could be recognized within each of these centers. LIBS data sets were collected in two different spectral bands (350±130 nm and 690±115 nm) using a Nd:YAG 1064 nm laser operated at ~23 mJ, a Czerny-Turner spectrograph with 0.2-0.3 nm spectral resolution and a high performance imaging charge couple device (ICCD) detector. Classification of the samples was performed using partial least-squares discriminant analysis (PLSDA), a common chemometric technique for performing statistical regression on high-dimensional data. Discrimination of samples from the Coso Volcanic Field, Bodie Hills, and other major obsidian areas in north-central California was possible with an accuracy of greater than 90% using either spectral band.

Electrochemical impedance spectroscopy-a simple method for the characterization of polymer inclusion membranes containing aliquat 336.

Electrochemical impedance spectroscopy (EIS) has been used to estimate the non-frequency dependent (static) dielectric constants of base polymers such as poly(vinyl chloride) (PVC), cellulose triacetate (CTA) and polystyrene (PS). Polymer inclusion membranes (PIMs) containing different amounts of PVC or CTA, along with the room temperature ionic liquid Aliquat 336 and plasticizers such as trisbutoxyethyl phosphate (TBEP), dioctyl sebecate (DOS) and 2-nitrophenyloctyl ether (NPOE) have been investigated. In this study, the complex and abstract method of EIS has been applied in a simple and easy to use way, so as to make the method accessible to membrane scientists and engineers who may not possess the detailed knowledge of electrochemistry and interfacial science needed for a rigorous interpretation of EIS results. The EIS data reported herein are internally consistent with a percolation threshold in the dielectric constant at high concentrations of Aliquat 336, which illustrates the suitability of the EIS technique since membrane percolation with ion exchangers is a well-known phenomenon.

Exposure of the gill epithelial cells of larval lampreys to an ion-deficient environment: a stereological study.

Three kinds of epithelial cells comprise the surfaces of the gill filaments and lamellae of larval lampreys (ammocoetes): ammocoete mitochondria-rich cells (AMRCs), intercalated mitochondria-rich cells (IMRCs) and pavement cells. Selected characteristics of these cell types in ammocoetes of Geotria australis held in distilled water and in 10% sea water were compared using an ultrastructural stereological approach to determine which of those cell type(s) respond to exposure to an ion-deficient environment in a manner that indicates that they are involved in ion uptake. Particular focus was placed on the enigmatic AMRC, which comprises ca 60% of the cells and contains numerous mitochondria. The mean percentage contributions of both AMRCs and pavement cells to the total number of the three cell types in the two experimental groups were not significantly different, whereas that of IMRCs was >7% in distilled water and <1% in 10% sea water (P < 0.001). Furthermore, the mean apical surface areas of neither AMRCs nor pavement cells differed significantly between the two experimental groups, whereas that of IMRCs was nearly 3-fold greater in distilled water than in 10% sea water. The volume densities and size of mitochondria in AMRCs did not differ between the two exposure regimes. The above comparisons provide no indications that the uptake of Na(+) and Cl(-) in the gill epithelium of ammocoetes involves either the AMRC or pavement cell but, when considered in conjunction with data on ion-transporting cells in other vertebrates, they are consistent with the conclusion that the IMRC plays a crucial role in this process.