Updates on antifungal pharmacotherapy in elasmobranchs: pharmacokinetics of 4 mg/kg voriconazole after IM and IV administration in undulate skates (Raja undulata) maintained under human care.

Introduction: Fungal diseases are frequently associated with elevated mortality rates in elasmobranchs. Currently, there is a notable absence of scientifically validated therapeutic medications that can ensure both effectiveness and safety when administered to this group of animals. The empirical prescription of azole antifungal agents, particularly voriconazole, has been posited as a potentially efficacious treatment approach for addressing most common mycoses in sharks and rays. However, there are still no published pharmacokinetic studies supporting its use in elasmobranchs and there is a lack of scientific base for its utilization in elasmobranchs. Methods: For this study, voriconazole was administered intravenously (IV) and intramuscularly (IM), at a single dose of 4 mg/kg to six adult undulate skates (Raja undulata). A washout period of 8 weeks was left between each route of administration. Blood samples were collected both before and at ten predetermined intervals after each dosing (0.25, 0.5, 1, 1.5, 2, 4, 8, 12, 24, and 36 h after drug administration). Plasma concentrations were quantified using a validated high-performance liquid chromatography method, and pharmacokinetic (PK) data was analyzed through non-compartmental methods. Results: The mean extrapolated concentration at 0 h (C0) after IV administration was 27.19 ± 7.15 µg/mL and the mean peak plasma concentrations (Cmax) ± SEM after IM administration resulted 2.98 ± 0.28 µg/mL at a mean time to maximum concentration (T max) of 1.33 ± 0.17 h. Terminal half-lives were calculated and resulted 11.18 ± 1.32 h for IV injections and 9.59 ± 1.38 h for IM injections. The area under the curve extrapolated to infinity was determined as 58.14 ± 2.79 h·µg/ml following IV injections and 37.60 ± 6.67 h·µg/ml following IM injections. The IM-administered voriconazole exhibited a mean absolute bioavailability of 64.67 ± 11.47%. Discussion: These discoveries provide backing for the possible application of voriconazole through the intramuscular route in undulate skates and support using lower dosage regimens compared to those required for oral administration, emphasizing the importance of conducting further pharmacokinetic studies with antifungals in elasmobranchs.

Linking extinction risk to the economic and nutritional value of sharks in small-scale fisheries.

To achieve sustainable shark fisheries, it is key to understand not only the biological drivers and environmental consequences of overfishing, but also the social and economic drivers of fisher behavior. The extinction risk of sharks is highest in coastal tropical waters, where small-scale fisheries are most prevalent. Small-scale fisheries provide a critical source of economic and nutritional security to coastal communities, and these fishers are among the most vulnerable social and economic groups. We used Kenya's and Zanzibar's small-scale shark fisheries, which are illustrative of the many data-poor, small-scale shark fisheries worldwide, as case studies to explore the relationship between extinction risk and the economic and nutritional value of sharks. To achieve this, we combined existing data on shark landings, extinction risk, and nutritional value with sales data at 16 key landing sites and information from interviews with 476 fishers. Shark fisheries were an important source of economic and nutritional security, valued at >US$4 million annually and providing enough nutrition for tens of thousands of people. Economically and nutritionally, catches were dominated by threatened species (72.7% and 64.6-89.7%, respectively). The most economically valuable species were large and slow to reproduce (e.g. mobulid rays, wedgefish, and bull, silky, and mako sharks) and therefore more likely to be threatened with extinction. Given the financial incentive and intensive fishing pressure, small-scale fisheries are undoubtedly major contributors to the decline of threatened coastal shark species. In the absence of effective fisheries management and enforcement, we argue that within small-scale fisheries the conditions exist for an economically incentivized feedback loop in which vulnerable fishers are driven to persistently overfish vulnerable and declining shark species. To protect these species from extinction, this feedback loop must be broken.

Conexión entre el riesgo de extinción y el valor nutricional de los tiburones en las pesquerías a pequeña escala Resumen Para lograr la sustentabilidad de las pesquerías de tiburones se deben entender los factores ecológicos y las consecuencias ambientales de la sobrepesca, así como los factores sociales y económicos del comportamiento del pescador. El riesgo de extinción de los tiburones es mucho mayor en las aguas tropicales costeras, en donde son más frecuentes las pesquerías a pequeña escala. Las pesquerías a pequeña escala, que además se encuentran entre los grupos con mayor vulnerabilidad social y económica, proporcionan una fuente importante de seguridad económica y nutricional para las comunidades costeras. Usamos las pesquerías de Kenia y Zanzíbar, las cuales representan muy bien a muchas de las pequeñas pesquerías de tiburones con deficiencia de datos, como estudios de caso para explorar la relación entre el riesgo de extinción y el valor económico y nutricional de los tiburones. Para lograr esto, combinamos los datos ya existentes de desembarques de tiburones, riesgo de extinción y valor nutricional con la información de ventas en 16 sitios clave de desembarque e información de las entrevistas a 476 pescadores. Las pesquerías de tiburones son una fuente importante de seguridad alimentaria y económica, valorada en más de US$4 millones anuales y que proporciona suficiente alimentación para miles de personas. En cuanto a la economía y la alimentación, las capturas estuvieron dominadas por especies amenazadas (72.7% y 64.6­89.7%, respectivamente). Las especies con mayor valor económico eran aquellas de gran tamaño y lenta reproducción, y, por lo tanto, con mayor probabilidad de estar en peligro de extinción. A causa del incentivo económico y la presión intensa de pesca, las pesquerías pequeñas sin duda son uno de los principales contribuyentes a la declinación de especies amenazadas de tiburones en las costas. Ya que no hay una aplicación ni un manejo efectivos de las pesquerías, argumentamos que en las pequeñas pesquerías existen las condiciones para un bucle de retroalimentación con incentivación económica en el que los pescadores vulnerables con frecuencia necesitan sobre pescar las especies de tiburones vulnerables y en declinación. Para proteger a estas especies de la extinción, este bucle de retroalimentación debe romperse.

Mercury bioaccumulation and its relationship with trophic biomarkers in a Mediterranean elasmobranch mesopredator.

Human activity has led to increased concentrations of mercury (Hg) in the world's oceans. Mercury can bioaccumulate and biomagnify in animal tissues via trophic transfer, thus, becoming most pronounced in larger and older predators. Here, we measured Hg concentrations and their relationship with stable isotopes-based proxies of trophic level (δ13C and δ15N values) in multiple tissues of Mustelus spp. from the Mediterranean Sea. We found higher Hg concentrations in muscle than in liver and fin tissues. The relationship between Hg concentrations and δ15N values in muscle suggested repeated foraging for low trophic level and Hg-poor prey, and biomagnification of Hg at higher trophic levels. Seasonal variations in δ13C values could indicate shifts in primary production sources and/or in local prey availability. The HBVSe index suggested no risk to human health, however the safe meal limit recommendations are 4.5 and 2.2 portions per month for adults and children, respectively.

The effect of fishing-capture stress on the oxygen uptake rate and swimming activity of the holocephalan Callorhinchus milii.

Overfishing, capture mortality, and consequences following the release of surviving animals represent severe threats to chondrichthyans. Although holocephalans are common bycaught and discarded species, other than postrelease mortality, little is known of fishing capture stress impacts. The stress response elicited after capture, essential to increase survival chances, is energetically demanding and affects the amount of energy available for other biological activities, with potential long-term impairments. We measured the effect of 30-min simulated gillnet capture on oxygen uptake rate (MO2 ), a proxy for metabolic rate and energy use, on recovery pattern, and on swimming activity of elephant fish (Callorhinchus milii). Immediately after simulated capture, Active and Inactive MO2 , measured during swimming and resting periods, respectively, were 27.5% and 43.1% lower than precapture values. This metabolic decline is likely an adaptation for reducing the energy allocated to non-essential activities, thus preserving it to sustain the stress response and processes essential for immediate survival. Supporting this, after gillnet capture, animals decreased their swimming time by 26.6%, probably due to a reduction in the energy allocated to movement. After 7 days, swimming activity and both Inactive MO2 and Active MO2 returned to precapture values. Although metabolic decline may enhance survival chances, the associated decreased swimming activity might increase predation risk and slow the physiological recovery after a fishing event. Moreover, some of the activities involved in Inactive MO2 are fundamental for life maintenance and therefore its depression after a capture event might have long-term repercussions for life sustenance and health.

Heavy metal concentrations in sharks, rays and chimaeras from the western Mediterranean Sea.

The potential bioaccumulation of pollutants, such as heavy metals, may pose a threat to the western Mediterranean chondrichthyans and human consumers. Therefore, the first extensive assessment of cadmium (Cd), lead (Pb), and copper (Cu) concentrations in the muscle tissue of 17 species of sharks, rays, and chimaeras in this region was conducted via Microwave Assisted Extraction (MAE) and Graphite Furnace Atomic Absorption Spectrometry (GFAAS). Significant differences between species were observed, particularly related to the rabbit fish (Chimaera monstrosa) and the velvet belly lantern shark (Etmopterus spinax), which exceeded the European Union (EU) Commission Regulation 2023/915 threshold of Cd. Overall, heavy metal concentrations correlated negatively with size and trophic level but positively with depth. Although the consumption of these species may entail minimal risk to adult humans, caution is advised, especially for children. These findings are important due to the widespread consumption of chondrichthyans in many western Mediterranean regions.

First record of partial albinism in the critically endangered Angelshark (Squatina squatina) (Linnaeus, 1758).

We report the first case of partial albinism in the Critically Endangered angelshark, Squatina squatina. The encounter with this specimen took place while SCUBA diving on the beach of Tufia, located on the east coast of the island of Gran Canaria on 2 April 2021. This is also the first confirmed finding of an albino elasmobranch specimen in the Canary Island archipelago.

Influence of female reproductive state and of fishing-capture stress on the oxygen uptake rate of a viviparous elasmobranch.

In animals discarded after a fishing capture event, the elicited stress response necessary to ensure their survival is energetically costly. This energy is diverted from other important biological activities, including growth and reproduction, possibly impairing them. Given that elasmobranchs are among the most threatened vertebrate groups, estimating capture-induced energetic changes and comparing these variations to the energy requirements of pregnancy maintenance is necessary. In pregnant southern fiddler rays (Trygonorrhina dumerilii), we measured changes in oxygen uptake rate (MO2 ; a proxy for metabolic rate and energy usage) in response to trawling simulation and air exposure, and estimated the oxygen requirements of sustaining late-term pregnancy and embryos. MO2 was measured in pregnant females, before (prestress MO2 ) and after trawling simulation (after-capture MO2 ), and again after females gave birth (postpartum MO2 ). After-capture MO2 was 31.7% lower than MO2 measured in minimally stressed females, suggesting a reduction in energy expenditure. This reduction is likely triggered by an initially excessive energetic investment in the stress response, and is aimed at shutting down nonessential activities to redirect energy to processes fundamental for survival. Prestress MO2 was 78.5% higher than postpartum MO2 . Capture simulation decreased MO2 to values similar to those observed postpartum, suggesting a capture-induced reduction in oxygen and energy allocation to pregnancy and embryonic respiration, which could be associated with reproductive impairments. These data, by better estimating the impact of capture and discard on energetic requirements and reproductive fitness, may support the introduction of area and/or seasonal closures to fishing.

Trace elements in edible tissues of elasmobranchs from the North Aegean Sea (Eastern Mediterranean) and potential risks from consumption.

Trace elements have the potential to bioaccumulate in marine organisms and to biomagnify towards the upper levels of marine trophic webs, resulting in a range of negative effects on organisms. Elasmobranchs are highly susceptible to bioaccumulation of trace metals, while their consumption by humans is increasing worldwide. Therefore, it is important to monitor the trace metal content in the edible tissues of elasmobranchs. This work reveals the content of 12 trace metals in the edible tissues of 10 elasmobranch species caught in Greek waters. Levels above the permissible limits for Hg and Pb were found in some species, while analysis of the lifetime consumption risk for adults and children using the Target Hazard Quotient (THQ), revealed a high risk for two of the most toxic substances on the priority list for substances, namely As and Hg. These are preliminary results, and further research is required to understand better the issue.

Conservation benefits of a marine protected area on South African chondrichthyans.

Chondrichthyans are threatened worldwide due to their life-history traits combined with a plethora of anthropogenic impacts that are causing populations to collapse. Marine Protected Areas (MPAs) are a conservation option, but their efficacy for chondrichthyans is still unclear. Conservation efforts might be challenging especially in developing countries, due to a lack of resources and monitoring and limited data and stakeholder support. Here Baited Remote Underwater Stereo-Video systems (stereo-BRUVs) were deployed inside and outside a small partially protected MPA (Robberg MPA, Western Cape, South Africa) to assess the status of cartilaginous fishes' assemblages and to investigate the potential benefits derived from the presence of a marine reserve. Overall, 19 chondrichthyan species in 11 different families were observed. Chondrichthyans were observed in 78.5% of the sites and, of these, 89.7% of the MPA sites showed at least one chondrichthyan, while only in the 67.5% of surrounding exploited sites a cartilaginous fish was sighted. The presence of the MPA had a significant effect on the relative abundance of batoids, threatened species and local endemics, with more observations inside the MPA than outside, indicating the potential benefit of marine reserves on species that are more vulnerable to fishing pressure. Relative abundance was generally higher inside the bay than in the exposed area, and both relative abundance and species richness decreased significantly with depth. The analysis of the body length showed that the 35.5% of species had an average body length below maturity length, indicating that the area might be used as nursery ground for different species. This study provides evidence that MPAs, even though small and partially protected, can provide benefits for chondrichthyans, specifically to threatened species, endemic species and lesser-known species. Importantly, different environmental parameters must be considered to maximize the benefits an MPA can provide.

Diverse stem-chondrichthyan oral structures and evidence for an independently acquired acanthodid dentition.

The teeth of sharks famously form a series of transversely organized files with a conveyor-belt replacement that are borne directly on the jaw cartilages, in contrast to the dermal plate-borne dentition of bony fishes that undergoes site-specific replacement. A major obstacle in understanding how this system evolved is the poorly understood relationships of the earliest chondrichthyans and the profusion of morphologically and terminologically diverse bones, cartilages, splints and whorls that they possess. Here, we use tomographic methods to investigate mandibular structures in several early branching 'acanthodian'-grade stem-chondrichthyans. We show that the dentigerous jaw bones of disparate genera of ischnacanthids are united by a common construction, being growing bones with non-shedding dentition. Mandibular splints, which support the ventro-lateral edge of the Meckel's cartilage in some taxa, are formed from dermal bone and may be an acanthodid synapomorphy. We demonstrate that the teeth of Acanthodopsis are borne directly on the mandibular cartilage and that this taxon is deeply nested within an edentulous radiation, representing an unexpected independent origin of teeth. Many or even all of the range of unusual oral structures may be apomorphic, but they should nonetheless be considered when building hypotheses of tooth and jaw evolution, both in chondrichthyans and more broadly.

The erroneous chondrichthyan egg case assignments from the Devonian: implications for the knowledge on the evolution of the reproductive strategy within chondrichthyans.

Spiraxis interstrialis, and its junior synonym Fayolia mourloni, an uppermost Famennian (Upper Devonian) fossil first described as algae and subsequently interpreted as the oldest known chondrichthyan egg case, is reinvestigated based on the discovery of several additional specimens in Belgian collections. New data, in particular from micro-CT imaging, allow to refute S. interstrialis, and by extension also Spiraxis major (the type species of Spiraxis Newberry, non Adams) and Spiraxis randalli from the Famennian of New York and Pennsylvania, as chondrichthyan egg cases. Alternative interpretations of these enigmatic helicoidal fossils are discussed. The first occurrence of oviparity in the fossil record of chondrichthyans is thus not as old as previously thought and is close to the first occurrence of viviparity in this group, both being recognised now in the Mississippian. The question of which of both conditions is plesiomorphic within chondrichthyans, and more widely within vertebrates, is discussed. Also, the presence of the genus Spiraxis in both the USA (east coast) and Belgium reinforces the strong faunal resemblance already observed in both palaeogeographical areas. It suggests important faunal exchanges between these regions of the Euramerica landmass during the Famennian.

Persistent organic pollutants and chlorpyrifos in the cockfish Callorhinchus callorynchus (Holocephali: Callorhynchidae) from Argentine coastal waters: Influence of sex and maturity.

Organic contaminants are of great environmental concern due to their negative impacts on coastal ecosystems, especially on highly vulnerable species as chondrichthyans. Accumulation of organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs) and chlorpyrifos was assessed in muscle, gonads, and liver of the cockfish Callorhinchus callorynchus for both sexes and maturity stages. This species has a wide distribution in South Atlantic Ocean and it is an important resource for coastal Argentine fisheries. Pollutants were quantified by gas chromatography with electron capture detector and the distribution pattern found in C. callorynchus was the following: (∑OCPs+chlorpyrifos) > ∑PCBs>∑PBDEs. Endosulfan was predominant among OCP groups, penta-CBs and hexaCBs among PCBs and BDE 47, 66, 99, and 100 in PBDE group. The highest levels were found in liver followed by muscle and gonads. The highest percentage of lipids was also observed in the liver. Moreover, the concentrations of ∑OCPs and ∑PCBs in C. callorynchus liver varied with maturity stage, and ∑OCPs also with sexes. Females presented higher values than males, and mature individuals showed higher concentrations than immature ones, according with biological parameters such as age, sex, maturity stage, metabolic and redistribution processes and habitat use influence. These results indicate that C. callorynchus reflects a historical and recent contamination in their tissues, and therefore, especially females, becomes as a good biomonitor of these pollutants in the marine environment. To our knowledge, this work represents one of the few investigations on the occurrence of POPs and chlorpyrifos in chondrichtyans from South Atlantic Ocean; therefore more research is mandatory for an adequate management and conservation of existing fisheries and aquatic resources.

Ancient fishes and the functional evolution of the corticosteroid stress response in vertebrates.

The neuroendocrine mechanism underlying stress responses in vertebrates is hypothesized to be highly conserved and evolutionarily ancient. Indeed, elements of this mechanism, from the brain to steroidogenic tissue, are present in all vertebrate groups; yet, evidence of the function and even identity of some elements of the hypothalamus-pituitary-adrenal/interrenal (HPA/I) axis is equivocal among the most basal vertebrates. The purpose of this review is to discuss the functional evolution of the HPA/I axis in vertebrates with a focus on our understanding of this neuroendocrine mechanism in the most ancient vertebrates: the agnathan (i.e., hagfish and lamprey) and chondrichthyan fishes (i.e., sharks, rays, and chimeras). A review of the current literature presents evidence of a conserved HPA/I axis in jawed vertebrates (i.e., gnathostomes); yet, available data in jawless (i.e., agnathan) and chondrichthyan fishes are limited. Neuroendocrine regulation of corticosteroidogenesis in agnathans and chondrichthyans appears to function through similar pathways as in bony fishes and tetrapods; however, key elements have yet to be identified and the involvement of melanotropins and gonadotropin-releasing hormone in the stress axis in these ancient fishes warrants further investigation. Further, the identities of physiological glucocorticoids are uncertain in hagfishes, chondrichthyans, and even coelacanths. Resolving these and other knowledge gaps in the stress response of ancient fishes will be significant for advancing knowledge of the evolutionary origins of the vertebrate stress response.

Structure of the paraplacenta and the yolk sac placenta of the viviparous Australian sharpnose shark, Rhizoprionodon taylori.

INTRODUCTION: Viviparity (live-birth) has evolved from oviparity (egg-laying) multiple times in sharks. While most transitions from oviparity to viviparity have resulted in non-placental forms of viviparity, some sharks develop a yolk sac placenta during pregnancy. The Australian sharpnose shark (Rhizoprionodon taylori) is a placental species that suspends embryonic development in a diapause for most of pregnancy. METHODS: To identify structures involved in supporting rapid embryonic growth in late pregnancy, we examined uterine and placental morphology by light and electron microscopy. RESULTS: Paraplacental uterine regions have morphological specialisations consistent with secretion and fluid transport between uterine tissues and the lumen. Uterine secretions in the lumen may be absorbed by the outgrowths on the embryonic umbilical cord ('appendiculae'), which are densely covered by microvilli. The placenta consists of uterine villi that interdigitate with the yolk sac and enhance the surface area available for fetomaternal exchange. The yolk sac does not invade the uterine epithelium, and the egg capsule remains intact at the placental interface, separating maternal and fetal tissues. Some placental uterine epithelial cells are secretory, and endocytic vesicles in the opposing yolk sac ectodermal cells suggest that nutrient transport is by histotrophic uterine secretion followed by fetal absorption. Respiratory gases, water and possibly small nutrients likely diffuse across the placenta, where maternal and fetal blood vessels are ~2 µm apart. DISCUSSION: Placental structure in R. taylori is similar to most other sharks, but there are differences in cellular structures between species that may indicate species-specific placental transport mechanisms.

Architectural and ultrastructural features of tessellated calcified cartilage in modern and extinct chondrichthyan fishes.

Tessellated calcified cartilage (TCC) is a distinctive kind of biomineralized perichondral tissue found in many modern and extinct chondrichthyans (sharks, rays, chimaeroids and their extinct allies). Customarily, this feature has been treated somewhat superficially in phylogenetic analyses, often as a single "defining" character of a chondrichthyan clade. TCC is actually a complex hard tissue with numerous distinctive attributes, but its use as a character complex for phylogenetic analysis has not yet been optimized. This study attempts to improve this situation by presenting new terminology for certain aspects of tesseral architecture, including single-monolayered, multiple-monolayered, polylayered and voussoir tesserae; new histological data, including thin sections of TCC in several Palaeozoic taxa, and new proposals for ways in which various characters and states (many of which are defined here for the first time) could be applied in future phylogenetic analyses of chondrichthyan fishes. It can be concluded that many, but not all, of the unique attributes of modern TCC evolved by the Early Devonian (ca. 400 before present (bp)). The globular calcified cartilage reported in Silurian sinacanthids and the so-called subtessellated perichondral biomineralization (with irregular and ill-defined geometries of a layer or layers of calcified cartilage blocks) of certain extinct "acanthodians" (e.g., Climatius, Ischnacanthus, Cheiracanthus) could represent evolutionary precursors of TCC, which seems to characterize only part of the chondrichthyan total group. It is hypothesized that heavily biomineralized "layer-cake" TCC in certain Palaeozoic chondrichthyans perhaps served a dual physiological role, as a phosphate sink and in providing increased skeletal density in very large (>7 m) Devonian-Permian marine sharks such as ctenacanths and as an adaptation to calcium-deficient environments among Permo-Carboniferous non-marine sharks such as xenacanths. By contrast, the equivalent tissue in modern elasmobranchs probably serves only to reinforce regions of cartilage (mostly in the jaws) subjected to high loading. It is also noted that much of the variation observed in tesseral architecture (including localized remodelling), ultrastructure and histology in modern and extinct chondrichthyans is confined to the perichondrally facing cap zone (where Type-1 collagen matrix predominates in modern TCC), whereas the main body of the tessera (where Type-2 collagen matrix predominates) exhibits comparatively little evidence of remodelling and histological or structural variation.

Comparative anatomy of the fin muscles of non-sarcopterygian fishes, with notes on homology and evolution.

Limited gross anatomical information about the muscles of fins, in particular those of the median fins, creates substantial gaps in the comparative anatomy, homologies, and evolution of these muscles across fishes. The scarcity of data also makes it difficult to interpret results obtained in developmental studies done in model organisms, such as zebrafish. To overcome these gaps, we provide descriptions of the configuration of all appendicular muscles of Amia, median fins of Polypterus, and the dorsal and anal fins of Lepisosteus and Chondrostei. The musculature of other species, including sharks and sturgeons, is also revised. We describe muscles that were previously overlooked, report sexual dimorphism in the muscles of the anal fin of Polypterus, and reveal muscle variations within Polypterus males. Species dissected for the present study thus represent all major non-sarcopterygian extant clades of gnathostomes, i.e. Chondrichthyes, Polypteriformes, Chondrostei, Lepisosteiformes, Amiiformes and Teleostei. Moreover, we compare our observations with the relatively few works that have provided information about muscles of at least some fins of these taxa in order to provide a broad discussion on - and detailed schemes showing - the major evolutionary patterns within the appendicular musculature of these fishes. Such discussion provides an opportunity for a more comprehensive understanding of appendicular evolution and fish evolution in particular and of gnathostome and morphological evolution in general.

The importance of a single amino acid substitution in reduced red blood cell carbonic anhydrase function of early-diverging fish.

In most vertebrates, red blood cell carbonic anhydrase (RBC CA) plays a critical role in carbon dioxide (CO2) transport and excretion across epithelial tissues. Many early-diverging fishes (e.g., hagfish and chondrichthyans) are unique in possessing plasma-accessible membrane-bound CA-IV in the gills, allowing some CO2 excretion to occur without involvement from the RBCs. However, implications of this on RBC CA function are unclear. Through homology cloning techniques, we identified the putative protein sequences for RBC CA from nine early-diverging species. In all cases, these sequences contained a modification of the proton shuttle residue His-64, and activity measurements from three early-diverging fish demonstrated significantly reduced CA activity. Site-directed mutagenesis was used to restore the His-64 proton shuttle, which significantly increased RBC CA activity, clearly illustrating the functional significance of His-64 in fish red blood cell CA activity. Bayesian analyses of 55 vertebrate cytoplasmic CA isozymes suggested that independent evolutionary events led to the modification of His-64 and thus reduced CA activity in hagfish and chondrichthyans. Additionally, in early-diverging fish that possess branchial CA-IV, there is an absence of His-64 in RBC CAs and the absence of the Root effect [where a reduction in pH reduces hemoglobin's capacity to bind with oxygen (O2)]. Taken together, these data indicate that low-activity RBC CA may be present in all fish with branchial CA-IV, and that the high-activity RBC CA seen in most teleosts may have evolved in conjunction with enhanced hemoglobin pH sensitivity.

Redescription and First Nucleotide Sequences of Rhinobatonchocotyle pacifica Oliva & Luque, 1995 (Monogenea: Hexabothriidae), a Parasite of Pseudobatos planiceps (Garman, 1880) (Rhinopristiformes: Rhinobatidae) from Peru.

INTRODUCTION: Specimens of Rhinobatonchocotyle pacifica Oliva & Luque, 1995 (Hexabothriidae) were collected from the gill filaments of Pacific guitarfish, Pseudobatos planiceps (Garman, 1880) (Rhinopristiformes: Rhinobatidae), from off the coast of Chorrillos, Lima, Peru. Since the original description of R. pacifica was incomplete, we herein redescribe R. pacifica based on re-examination of the holotype, paratypes, and newly collected voucher specimens. Nucleotide sequences (18S, 28S) for R. pacifica are presented for the first time, and a phylogenetic analysis conducted. MATERIALS AND METHODS: Monogeneans were examined with light microscopy and partial sequences of 18S and 28S rDNA genes were obtained. RESULTS: The following taxonomically important morphological features are described: presence of seminal receptacle, spiniform papillae within oral cavity, haptoral sucker sclerites and anchors, hook surface striations, muscular pad associated with each haptoral sucker sclerite (except sclerite 1), muscular bulbs in appendix suckers, and oötype lacking longitudinal rows of large cells (oötype not côtelé). The genital complex is detailed. The diagnosis of Rhinobatonchocotyle is emended to include species with or without seminal receptacle. Representative nucleotide sequences of partial 18S and 28S rDNA genes of R. pacifica were obtained for the first time and used for reconstructing the phylogeny of Hexabothriidae. CONCLUSION: The phylogenetic reconstruction confirmed the species within the Hexabothriidae.

Observations of two deep-sea chimaeroids in Icelandic waters.

Long-term survey time series repeated occurrence of the deep-sea chimaeroid species small-eyed rabbitfish Hydrolagus affinis and large-eyed rabbitfish H. mirabilis have been observed at same but few locations in Icelandic waters over 23 years. For H. affinis the catch at one specific station has consisted almost exclusively of mature males over the time period. Identification of potential aggregation locations is of vital importance to natural resource management because such aggregation may render these species vulnerable to potential fishing activities.

A Tournaisian (earliest Carboniferous) conglomerate-preserved non-marine faunal assemblage and its environmental and sedimentological context.

A conglomerate bed from the Tournaisian Ballagan Formation of Scotland preserves a rich array of vertebrate and other nonmarine fossils providing an insight into the wider ecosystem and paleoenvironment that existed during this pivotal stage of Earth history. It challenges hypotheses of a long-lasting post-extinction trough following the end-Devonian extinction event. The fauna recovered includes a wide size range of tetrapods, rhizodonts, and dipnoans, from tiny juveniles or small-bodied taxa up to large adults, and more than one taxon of each group is likely. Some fauna, such as actinopterygians and chondrichthyans, are rare as macrofauna but are better represented in the microfossil assemblage. The fauna provides evidence of the largest Carboniferous lungfish ever found. The specimens are preserved in a localized, poorly-sorted conglomerate which was deposited in the deepest part of a river channel, the youngest of a group of channels. In addition to the fossils (micro- and macro-), the conglomerate includes locally-derived clasts of paleosols and other distinctive elements of the surrounding floodplains. Charcoal fragments represent small woody axes and possible larger trunk tissue from arborescent pteridosperms. Preservation of the fossils indicates some aerial exposure prior to transport, with abrasion from rolling. The findings presented here contrast with other published trends in vertebrate size that are used to interpret a reduction in maximum sizes during the Tournaisian. The richness of the fauna runs counter to the assumption of a depauperate nonmarine fauna following the end-Devonian Hangenberg event, and charcoal content highlights the occurrence of fire, with the requisite levels of atmospheric oxygen during that stage.