Cesarian section and long-term outcomes for cownose rays (Rhinoptera bonasus).

Cownose rays (Rhinoptera bonasus) are schooling rays commonly displayed in large groups in public aquariums. They are long-lived, have an annual reproductive cycle, and readily breed in managed care with most pregnancies culminating with the unaided and successful birth of a single neonate. Occasionally, females are observed to have prolonged pregnancies or suffer dystocia during parturition and intervention via a cesarian section (C-section) is required to deliver the neonate. Monthly reproductive monitoring at Ripley's Aquarium of the Smokies using ultrasound to stage pregnancies allows for the prediction of anticipated due dates and guides the decision to assist with delivery. Recognizing when to assist birth and best practices for performing C-section are important for the reproductive health, sustainability, and longevity of this species in managed care. This report describes a surgical technique for C-section in cownose rays and includes short-term complications and long-term outcomes for females.

Comparative Tissue Identification and Characterization of Long Non-Coding RNAs in the Globally Distributed Blue Shark Prionace glauca.

Long non-coding RNAs (lncRNAs) are involved in numerous biological processes and serve crucial regulatory functions in both animals and plants. Nevertheless, there is limited understanding of lncRNAs and their patterns of expression and roles in sharks. In the current study, we systematically identified and characterized lncRNAs in the blue shark (Prionace glauca) from four tissues (liver, spleen, muscle, and kidney) using high-throughput sequencing and bioinformatics tools. A total of 21,932 high-confidence lncRNAs were identified, with 8984 and 3067 stably and tissue-specific expressed lncRNAs, respectively. In addition, a total of 45,007 differentially expressed (DE) lncRNAs were obtained among tissues, with kidney versus muscle having the largest numbers across tissues. DE lncRNAs trans target protein-coding genes were predicted, and functional gene ontology enrichment of these genes showed GO terms such as muscle system processes, cellular/metabolic processes, and stress and immune responses, all of which correspond with the specific biological functions of each tissue analyzed. These results advance our knowledge of lncRNAs in sharks and present novel data on tissue-specific lncRNAs, providing key information to support future functional shark investigations.

Novel use of deep neural networks on photographic identification of epaulette sharks (Hemiscyllium ocellatum) across life stages.

Photographic identification (photo ID) is an established method that is used to count animals and track individuals' movements. This method performs well with some species of elasmobranchs (i.e., sharks, skates, and rays) where individuals have distinctive skin patterns. However, the unique skin patterns used for ID must be stable through time to allow re-identification of individuals in future sampling events. More recently, artificial intelligence (AI) models have substantially decreased the labor-intensive process of matching photos in extensive photo ID libraries and increased the reliability of photo ID. Here, photo ID and AI are used for the first time to identify epaulette sharks (Hemiscyllium ocellatum) at different life stages for approximately 2 years. An AI model was developed to assess and compare the reliability of human-classified ID patterns in juvenile and neonate sharks. The model also tested the persistence of unique patterns in adult sharks. Results indicate that immature life stages are unreliable for pattern identification, using both human and AI approaches, due to the plasticity of these subadult growth forms. Mature sharks maintain their patterns through time and can be identified by AI models with approximately 86% accuracy. The approach outlined in this study has the potential of validating the stability of ID patterns through time; however, testing on wild populations and long-term datasets is needed. This study's novel deep neural network development strategy offers a streamlined and accessible framework for generating a reliable model from a small data set, without requiring high-performance computing. Since many photo ID studies commence with limited datasets and resources, this AI model presents practical solutions to such constraints. Overall, this approach has the potential to address challenges associated with long-term photo ID data sets and the application of AI for shark identification.

Life cycle and development of the marine leech Branchellion lobata (Hirudinea: Piscicolidae), from round stingrays, Urobatis halleri, from southern California.

During captivity, round stingrays, Urobatis halleri, became infected with the marine leech Branchellion lobata. When adult leeches were deprived of blood meal, they experienced a rapid decrease in body mass and did not survive beyond 25 days. If kept in aquaria with host rays, B. lobata fed frequently and soon produced cocoons, which were discovered adhered to sand grains. A single leech emerged from each cocoon (at ~ 21 days), and was either preserved for histology or molecular analysis, or monitored for development by introduction to new hosts in aquaria. Over a 74-day observation period, leeches grew from ~ 2 to 8 mm without becoming mature. Newly hatched leeches differed from adults in lacking branchiae and apparent pulsatile vesicles. The microbiome of the hatchlings was dominated by a specific, but undescribed, member of the gammaproteobacteria, also recovered previously from the adult leech microbiome. Raising B. lobata in captivity provided an opportunity to examine their reproductive strategy and early developmental process, adding to our limited knowledge of this common group of parasites.

Baseline polycyclic aromatic hydrocarbon maternal transfer data in Lesser Numbfish Narcine brasiliensis (Elasmobranchii: Batoidea) from an impacted estuary in Southeastern Brazil.

Maternal offloading of polycyclic aromatic hydrocarbons (PAHs) poses a significant exposure route for developing embryos, with implications for subsequent generations. Despite known developmental effects regarding fish physiology and behavior, maternal PAH transfer assessments in elasmobranchii are still lacking. This study investigated PAH contamination and maternal transfer in one female Lesser Numbfish (Narcine brasiliensis) electric ray and seven embryos for the first time. Naphthalene was identified as the predominant low molecular weight PAH, and dibenzo[a,h]anthracene was the most abundant high molecular weight compound. Most embryos exhibited some level of PAH exposure, with varying accumulation patterns potentially influenced by size, developmental stage, and yolk absorption rates. Further investigation is warranted to understand the impacts of PAH maternal offloading on elasmobranchii uterine contents and embryos.

Spiny dogfish, Squalus suckleyi, shows a good tolerance for hypoxia but need long recovery times.

Pacific spiny dogfish, Squalus suckleyi, move to shallow coastal waters during critical reproductive life stages and are thus at risk of encountering hypoxic events which occur more frequently in these areas. For effective conservation management, we need to fully understand the consequences of hypoxia on marine key species such as elasmobranchs. Because of their benthic life style, we hypothesized that S. suckleyi are hypoxia tolerant and able to efficiently regulate oxygen consumption, and that anaerobic metabolism is supported by a broad range of metabolites including ketones, fatty acids and amino acids. Therefore, we studied oxygen consumption rates, ventilation frequency and amplitude, blood gasses, acid-base regulation, and changes in plasma and tissue metabolites during progressive hypoxia. Our results show that critical oxygen levels (P crit) where oxyregulation is lost were indeed low (18.1% air saturation or 28.5 Torr at 13°C). However, many dogfish behaved as oxyconformers rather than oxyregulators. Arterial blood PO2 levels mostly decreased linearly with decreasing environmental PO2. Blood gases and acid-base status were dependent on open versus closed respirometry but in both set-ups ventilation frequency increased. Hypoxia below Pcrit resulted in an up-regulation of anaerobic glycolysis, as evidenced by increased lactate levels in all tissues except brain. Elasmobranchs typically rely on ketone bodies as oxidative substrates, and decreased concentrations of acetoacetate and ß-hydroxybutyrate were observed in white muscle of hypoxic and/or recovering fish. Furthermore, reductions in isoleucine, glutamate, glutamine and other amino acids were observed. After 6 hours of normoxic recovery, changes persisted and only lactate returned to normal in most tissues. This emphasizes the importance of using suitable bioindicators adjusted to preferred metabolic pathways of the target species in conservation physiology. We conclude that Pacific spiny dogfish can tolerate severe transient hypoxic events, but recovery is slow and negative impacts can be expected when hypoxia persists.

The complete mitochondrial genome of the imperiled Bullnose ray Myliobatis freminvillei (Myliobatiformes: Myliobatidae) with comments on its phylogenetic position and claims of diversifying selection affecting protein coding genes in a closely related species.

The Bullnose ray Myliobatis freminvillei is a bentho-pelagic eagle ray that inhabits the north Gulf of Mexico and the Western Atlantic Ocean Coast, discontinuously, from Massachusetts, USA to Buenos Aires, Argentina. Myliobatis freminvillei is currently listed as vulnerable by the 2019 IUCN Red List of Threatened Species given that it is often captured as bycatch by artisanal and commercial fisheries, along the coasts of Argentina, Brazil, and Venezuela. This study, for the first time, assembled and characterized the mitochondrial genome of M. fremnvillei. The A+T rich mitochondrial genome of M. fremnvillei is 18,356 bp long and encodes 22 transfer RNA genes (tRNA), 2 ribosomal RNA genes (12S ribosomal RNA and 16S ribosomal RNA), 13 protein coding genes (PCGs), and also contains a non coding control region 2,617 bp long. Nonsynonomous codon usage with a preference for A+T rich codons was observed in all 13 PCGs. Leu (CTA), Ile (ATC), Phe (TTC), Thr (ACA), and Ala (GCC) were the most frequently used codons. Ka/Ks ratios estimated for all 13 PCGs exhibited values < 1, indicating strong purifying selection affecting all these genes. In contrast to the results of a previous study that claimed diversifying selective pressure in two mitochondrial PCGs of Mobula tarapacana, reanalysis of the Ka/Ks values for the same species indicated purifying selection in all 13 PCGs. Of the 22 tRNA genes, all have a cloverleaf secondary structure except tRNA-Ser1 which has a truncated dihydrouridine arm. In the control region, A+T rich microsatellites (n = 42) and short tandem repeats (n = 6) were identified, and the secondary structure of the same region contained numerous hairpin loops. Phylomitogenomic analyses supported the monophyletic status of the order Myliobatiformes and family Myliobatidae. The assembled mitochondrial genome will assist with conservation efforts in Myliobatis fremnvillei.

How hot is too hot? Thermal tolerance, performance, and preference in juvenile mangrove whiprays, Urogymnus granulatus.

Mangrove habitats can serve as nursery areas for sharks and rays. Such environments can be thermally dynamic and extreme; yet, the physiological and behavioural mechanisms sharks and rays use to exploit such habitats are understudied. This study aimed to define the thermal niche of juvenile mangrove whiprays, Urogymnus granulatus. First, temperature tolerance limits were determined via the critical thermal maximum (CTMax) and minimum (CTMin) of mangrove whiprays at summer acclimation temperatures (28 °C), which were 17.5 °C and 39.9 °C, respectively. Then, maximum and routine oxygen uptake rates (MO2max and MO2routine, respectively), post-exercise oxygen debt, and recovery were estimated at current (28 °C) and heatwave (32 °C) temperatures, revealing moderate temperature sensitivities (i.e., Q10) of 2.4 (MO2max) and 1.6 (MO2routine), but opposing effects on post-exercise oxygen uptake. Finally, body temperatures (Tb) of mangrove whiprays were recorded using external temperature loggers, and environmental temperatures (Te) were recorded using stationary temperature loggers moored in three habitat zones (mangrove, reef flat, and reef crest). As expected, environmental temperatures varied between sites depending on depth. Individual mangrove whiprays presented significantly lower Tb relative to Te during the hottest times of the day. Electivity analysis showed tagged individuals selected temperatures from 24.0 to 37.0 °C in habitats that ranged from 21.1 to 43.5 °C. These data demonstrate that mangrove whiprays employ thermotaxic behaviours and a thermally insensitive aerobic metabolism to thrive in thermally dynamic and extreme habitats. Tropical nursery areas may, therefore, offer important thermal refugia for young rays. However, these tropical nursery areas could become threatened by mangrove and coral habitat loss, and climate change.

The olfactory system of sharks and rays in numbers.

Cartilaginous fishes have large and elaborate olfactory organs, but only a small repertoire of olfactory receptor genes. Here, we quantitatively analyze the olfactory system of 21 species of sharks and rays, assessing many features of the olfactory organ (OOR) (number of primary lamellae, branches of the secondary folds, sensory surface area, and density and number of sensory neurons) and the olfactory bulb (OB) (number of neurons and non-neuronal cells), and estimate the ratio between the number of neurons in the two structures. We show that the number of lamellae in the OOR does not correlate with the sensory surface area, while the complexity of the lamellar shape does. The total number of olfactory receptor neurons ranges from 30.5 million to 4.3 billion and the total number of OB neurons from 1.5 to 90 million. The number of neurons in the olfactory epithelium is 16 to 158 times higher (median ratio is 46) than the number of neurons in the OB. These ratios considerably exceed those reported in mammals. High convergence from receptor neurons to neurons processing olfactory information, together with the remarkably small olfactory receptor repertoire, strongly suggests that the olfactory system of sharks and rays is well adapted to detect a limited number of odorants with high sensitivity.

Vulnerability of Eastern Tropical Pacific chondrichthyan fish to climate change.

Climate change is an environmental emergency threatening species and ecosystems globally. Oceans have absorbed about 90% of anthropogenic heat and 20%-30% of the carbon emissions, resulting in ocean warming, acidification, deoxygenation, changes in ocean stratification and nutrient availability, and more severe extreme events. Given predictions of further changes, there is a critical need to understand how marine species will be affected. Here, we used an integrated risk assessment framework to evaluate the vulnerability of 132 chondrichthyans in the Eastern Tropical Pacific (ETP) to the impacts of climate change. Taking a precautionary view, we found that almost a quarter (23%) of the ETP chondrichthyan species evaluated were highly vulnerable to climate change, and much of the rest (76%) were moderately vulnerable. Most of the highly vulnerable species are batoids (77%), and a large proportion (90%) are coastal or pelagic species that use coastal habitats as nurseries. Six species of batoids were highly vulnerable in all three components of the assessment (exposure, sensitivity and adaptive capacity). This assessment indicates that coastal species, particularly those relying on inshore nursery areas are the most vulnerable to climate change. Ocean warming, in combination with acidification and potential deoxygenation, will likely have widespread effects on ETP chondrichthyan species, but coastal species may also contend with changes in freshwater inputs, salinity, and sea level rise. This climate-related vulnerability is compounded by other anthropogenic factors, such as overfishing and habitat degradation already occurring in the region. Mitigating the impacts of climate change on ETP chondrichthyans involves a range of approaches that include addressing habitat degradation, sustainability of exploitation, and species-specific actions may be required for species at higher risk. The assessment also highlighted the need to further understand climate change's impacts on key ETP habitats and processes and identified knowledge gaps on ETP chondrichthyan species.

El cambio climático es una emergencia medioambiental que amenaza a especies y ecosistemas en todo el mundo. Los océanos han absorbido alrededor del 90% del calor antropogénico y entre el 20% y el 30% de las emisiones de carbono, lo que ha provocado su calentamiento, acidificación, desoxigenación, cambios en la estratificación de los océanos y en la disponibilidad de nutrientes, así como fenómenos extremos más pronunciados. Dadas las predicciones de cambios, hay una importante necesidad de entender cómo las especies marinas se verán afectadas. En este estudio utilizamos una Evaluación Integrada de Riesgos para evaluar la vulnerabilidad de 132 condrictios del Pacífico Tropical Oriental (PTO) a los impactos del cambio climático. Adoptando un enfoque preventivo, estimamos que la vulnerabilidad general al cambio climático es Alta para casi una cuarta parte (23%) de las especies de condrictios del PTO evaluadas y Moderada para gran parte del resto (76%). La mayoría de las especies altamente vulnerables son batoideos (77%), y una gran proporción de éstas (90%) son especies costeras o especies pelágicas que utilizan los hábitats costeros como áreas de crianza. Seis especies de batoideos tuvieron una vulnerabilidad Alta en los tres componentes de la evaluación. Esta evaluación indica que las especies costeras, en particular las que dependen de áreas de crianza costeras, son las más vulnerables al cambio climático. Es probable que el calentamiento de los océanos, junto con la acidificación y la posible desoxigenación, tenga efectos generalizados sobre las especies de condrictios del PTO, pero las especies costeras se verán también afectadas por los cambios en los aportes de agua dulce, la salinidad y el aumento del nivel del mar. Esta vulnerabilidad relacionada con el clima se ve agravada por otros factores antropogénicos que ya se están produciendo en la región, como la sobrepesca y la degradación del hábitat. La mitigación de los impactos del cambio climático sobre los condrictios del PTO implica medidas que incluyan abordar la degradación del hábitat y la sostenibilidad de la explotación pesquera, y acciones para las especies de mayor riesgo son necesarias. Esta evaluación también destaca la necesidad de comprender mejor los impactos del cambio climático en los hábitats y procesos clave del PTO y las lagunas de conocimiento identificadas en relación con las especies de condrictios del PTO.