Impact of human-associated Escherichia coli clonal groups in Antarctic pinnipeds: presence of ST73, ST95, ST141 and ST131

There is growing concern about the spreading of human microorganisms in relatively untouched ecosystems such as the Antarctic region. For this reason, three pinniped species (Leptonychotes weddellii, Mirounga leonina and Arctocephalus gazella) from the west coast of the Antartic Peninsula were analysed for the presence of Escherichia spp. with the recovery of 158 E. coli and three E. albertii isolates. From those, 23 harboured different eae variants (α1, β1, β2, ε1, θ1, κ, ο), including a bfpA-positive isolate (O49:H10-A-ST206, eae-k) classified as typical enteropathogenic E. coli. Noteworthy, 62 of the 158 E. coli isolates (39.2%) exhibited the ExPEC status and 27 (17.1%) belonged to sequence types (ST) frequently occurring among urinary/bacteremia ExPEC clones: ST12, ST73, ST95, ST131 and ST141. We found similarities >85% within the PFGE-macrorrestriction profiles of pinniped and human clinic O2:H6-B2-ST141 and O16:H5/O25b:H4-B2-ST131 isolates. The in silico analysis of ST131 Cplx genomes from the three pinnipeds (five O25:H4-ST131/PST43-fimH22-virotype D; one O16:H5-ST131/PST506-fimH41; one O25:H4-ST6252/PST9-fimH22-virotype D1) identified IncF and IncI1 plasmids and revealed high core-genome similarities between pinniped and human isolates (H22 and H41 subclones). This is the first study to demonstrate the worrisome presence of human-associated E. coli clonal groups, including ST131, in Antarctic pinnipeds.The sampling was funded by the Spanish Ministry of Science and Innovation (CGL-2005-25073-E/ANT and CTM2008-00570) and the Sea World & Bush Gardens Conservation Fund. Work at USC-LREC was supported by projects AGL2013-47852-R from the Ministerio de Economía y Competitividad (MINECO, Spain) and Fondo Europeo de Desarrollo Regional (FEDER); AGL2016-79343-R from the Agencia Estatal de Investigación (AEI, Spain) and FEDER; PI16/01477 from Plan Estatal de I + D + I 2013-2016, Instituto de Salud Carlos III (ISCIII), Subdirección General de Evaluación y Fomento de la Investigación, and FEDER; CN2012/303 and ED431C 2017/57 from the Consellería de Cultura, Educación e Ordenación Universitaria, (Xunta de Galicia) and FEDER. We would like to thank the military personnel at the Spanish Antarctic Base “Gabriel de Castilla” for their help and assistance and the Marine Technology Unit (CSIC), the Spanish Navy’s Oceanographic Research Ship “Las Palmas” and the Unidad de Tecnología Marítima (UTM, CSIC) for logistics and transport. We also express our gratitude to J. Castro-Urda, F.T. García-Moreno, C. Rengifo-Herrera, S. Rojo-Montejo, I. Ferre, V. Navarro, M. Gómez-Bautista, T. Alvaro-Alvarez and J. Coello-Pérez for their invaluable help in the sample collection. A. Mora acknowledges the Ministerio de Educación, Cultura y Deporte (Spain) for the mobility grant for teachers and researchers from the Programa Estatal de Promoción del Talento y su Empleabilidad, Plan Estatal de Investigación Científica y Técnica y de Innovación 2013–2016.S

Tension pneumothorax in small odontocetes

Pneumothorax, the accumulation of air in the pleural cavity, occurs when air enters the pleural space by the pleuro-cutaneous, pleuro-pulmonary, or pleuro-oesophageal-mediastinal route. Tension pneumothorax is an infrequent and severe form of pneumothorax where a positive pressure in the pleural space is built up during at least part of the respiratory cycle, with compression of both lungs and mediastinal vessels, and, if unilateral, with midline deviation towards the unaffected hemithorax. We describe 9 cases of tension pneumothorax in 3 species of small cetaceans (striped dolphin Stenella coeruleoalba, common dolphin Delphinus delphis, and common bottlenose dolphin Tursiops truncatus) from the western Mediterranean coast of Spain, and one case from a dolphinarium. Computed tomography (CT) imaging performed in 2 carcasses before necropsy showed lung compression, midline deviation, and pressure on the diaphragm, which was caudally displaced. Tension pneumothorax was recognized at necropsy by the presence of pressurized air in one of the hemithoraces. Seven of the pneumothorax cases were spontaneous (2 primary and 5 secondary to previous lung pathology). In the other 2 dolphins, the pneumothorax was traumatic, due to oesophageal-pleural perforation or rib fractures. We hypothesize that pneumothorax in dolphins is predominantly tensional because of their specific anatomical and physiological adaptations to marine life and the obligate exposure to extreme pressure changes as diving mammals.This research was conducted under Direcció General de Medi Natural i Biodiversitat Permit. We thank the Servei de Vida Silvestre Conselleria d’Agricultura, Medi Ambient, Canvi Climàtic i Desenvolupament Rural, Generalitat Valenciana for their support and permits. We thank Blanca Pérez and Aida Neira from the Servei de Diagnòstic de Patologia Veterinària, Universitat Autònoma de Barcelona. Our thanks to colleagues from the Marine Zoology Unit from University of Valencia, for their assistance and support in obtaining data from stranded cetaceans. We also thank ZOETIS Spain for supporting Fundación Oceanogràfic in veterinary diagnosis and stranding medical response. Funding Financial support for necropsies at UAB was provided by Direcció General de Medi Natural i Biodiversitat, Generalitat de Catalunya. Fundación Oceanogràfic has the support of the Fundación Biodiversidad of the Ministerio de Transición Ecológica y Reto Demográfico Challenge (MITECO) within the framework of the Recovery, Transformation and Resilience Plan (PRTR), financed by the European Union − NextGenerationEU.info:eu-repo/semantics/publishedVersio

Effect of temperature and cell concentration on dolphin (Tursiops truncatus) spermatozoa quality evaluated at different days of refrigeration

12 Pág.The use of cryopreserved dolphin spermatozoa facilitates the exchange of genetic material between aquatic parks and makes spermatozoa accessible to laboratories for studies to further our understanding of marine mammal reproduction. Sperm cryopreservation in the bottlenose dolphin (Tursiops truncatus) has been developed for the exchange of gametes within the ex situ population. The aim of this study was to develop an effective method for refrigeration of bottlenose dolphin spermatozoa diluted in a commercial extender (BTS). In Experiment 1, the effect of temperature (5 compared with 15 °C) on sperm quality was evaluated during 7 days of storage at 100 × 106 spermatozoa/ml. In Experiment 2, the effect of the storage concentration (100 × 106 compared with 20 × 106 spermatozoa/ml) on sperm quality was assessed during 7 days of storage at 5 °C. In Experiment 1, total motility (including % of rapid sperm) was greater at 5 than 15 °C. When the effect of storage concentration was evaluated (Experiment 2), total motility and ALH were greater at the higher storage concentration (100 × 106 spermatozoa/ml). For both experiments, values for viability, acrosome integrity, and normal morphology variables were consistent throughout the 7 days of refrigeration. In Experiment 3, a microbiological study was performed to evaluate the effect of the refrigeration temperature and days of storage on bacterial growth. The results of microbiological analysis indicated there was Staphylococcus aureus in some samples, however, there was no effect of temperature or days of refrigeration. In conclusion, bottlenose dolphin semen can be refrigerated for a short to medium period of storage and there is maintenance of functionality of sperm when stored at 100 × 106 spermatozoa/ml at 5 °C.There was not any specific grant from funding agencies in the public, commercial, or not-for-profit sectors obtained to conduct the research reported in this manuscript.Peer reviewe

B-esterase measurements and other blood related biomarkers in loggerhead sea turtles (Caretta caretta) as indicators of health status

9 pages, 4 figures, 4 tables, supplementary data https://doi.org/10.1016/j.scitotenv.2023.163040.-- Data availability: Data will be made available on requestThe loggerhead sea turtle (Caretta caretta) has been selected as sentinel species by the Marine Strategy Framework Directive (MSFD) descriptor 10 in relation to marine litter. In this, and other protected species, there is a need to develop conservative pollution biomarkers equally informative of chemical exposures to those traditionally carried out in metabolic organs, such as the liver. With this aim, plasma from turtles undergoing rehabilitation at the Fundació Oceanogràfic rescue centre (Arca del Mar) were selected and tested for B-esterase measurements. Hydrolysis rates of acetylcholinesterase (AChE), butyrylcholinesterase (BChE) and carboxylesterases (CEs) using four commercial substrates were undertaken on 191 plasma samples. Results indicated that acetylthiocholine was the most adequate substrate of cholinesterases and butyrate esters for CE measures. The correlation of these parameters with well-established blood biochemistry measurements was analysed. B-esterase measures in wild specimens were discussed in relation to age group, pathology on admission to the rescue centre and season; moreover, contrasts with long-term resident turtles were also made. Although this study provides baseline data on B-esterase measures in a large sample size for this species, more complementary information is still needed in terms of population genetics, chemical exposures, and in relation to other biochemical parameters before they can be confidently applied in wild specimens within the regulatory MSFDTo the institutional support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S). This work has received funding from CSIC Interdisciplinary Thematic Platform (PTI+) Interdisciplinary Platform for Sustainable Plastics towards a Circular Economy+ (PTI-SusPlast+Peer reviewe

Characterisation of plasmatic B-esterases in bottlenose dolphins (Tursiops truncatus) and their potential as biomarkers of xenobiotic chemical exposures

9 pages, 5 pages, 3 figures, supplementary data https://doi.org/10.1016/j.envpol.2022.120149.-- Data availability: Data will be made available on requestA total of 164 blood samples from 16 clinically healthy bottlenose dolphins (Tursiops truncatus), were obtained from an aquarium in Spain between 2019 and 2020, as part of their preventive medicine protocol. In addition to conventional haematological and biochemical analyses, plasmatic B-esterase activities were characterised to determine the potential application of such analyses in wild counterparts. The hydrolysis rates for the substrates of acetylcholinesterase (AChE), butyrylcholinesterase (BuChE) and carboxylesterase (CE) activity in plasma were measured, the last using two commercial substrates, p-nitrophenyl acetate (pNPA) and p-nitrophenyl butyrate (pNPB). Activity rates (mean ± SEM in nmol/min/mL plasma) were (in descending order): AChE (125.6 ± 3.8), pNPB-CE (65.0 ± 2.2), pNPA-CE (49.7 ± 1.1) and BuChE (12.8 ± 1.3). These values for dolphins are reported in here for the first time in this species. Additionally, the in vitro sensitivity of two B-esterases (AChE and pNPB-CE) to chemicals of environmental concern was determined, and the protective role of plasmatic albumin assessed. Out of the B-esterases measured in plasma of dolphin, AChE activity was more responsive in vitro to pesticides, while CEs had a low response to plastic additives, likely due to the protective presence of albumin. However, the clear in vitro interaction of these environmental chemicals with purified AChE from electric eels and recombinant human hCEs (hCE1 and hCE2) and albumin, predicts their impact in other tissues that require in vivo validation. A relationship between esterase-like activities and health parameters in terrestrial mammals has already been established. Thus, B-esterase measures could be easily included in marine mammal health assessment protocols for dolphins as well, once the relationship between these measures and the animal's fitness has been establishedTo the institutional support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S)Peer reviewe

Sinergia entre el Institut de Ciències del Mar (ICM-CSIC) y el Oceanogràfic de Valencia: un bloom de conocimiento sobre medusas

XXIV Bienal de la Real Sociedad Española de Historia Natural, La huella Humana en la Naturaleza, del 8 al 11 de septiembre de 2021, ValenciaLa cooperación potencia la cultura y aumenta la creatividad individual. Esta afirmación fue postulada por Charles Darwin en 1871 en su obra El Origen del Hombre. Este año se cumplen 150 años de su publicación y, coincidiendo con esta efeméride, se acaba de formalizar la cooperación entre el Institut de Ciències del Mar (ICM-CSIC) y el Oceanogràfic de Valencia con el objetivo de mejorar y ampliar el conocimiento sobre medusas. Esta sinergia pretende aumentar la investigación sobre estos cnidarios a través de la colaboración entre la empresa pública y privada. Ambas instituciones pueden sumar sus potencialidades; el ICM-CSIC aporta su gran experiencia científica y académica, mientras que el Oceanogràfic contribuye con la dilatada experiencia alcanzada en el manejo de las medusas y las extensas instalaciones dedicadas a estos animales. La exhibición de medusas del Oceanogràfic se inauguró en el año 2017, siendo por aquel entonces la exposición más grande dedicada a estos animales en Europa. En un principio, prácticamente todos los individuos de las 17 especies distintas que se llegaron a exhibir, fueron cedidos por otros acuarios. De hecho, el ICM-CSIC proporcionó ejemplares de algunas medusas mediterráneas para complementar la exposición. Actualmente, se ha alcanzado el mayor número de especies exhibidas (18 en total), siendo todas ellas reproducidas en el propio acuario de Valencia. La experiencia a lo largo de estos años ha permitido que el equipo de acuaristas haya alcanzado un nivel muy alto de conocimiento y manejo de estos animales, cerrando el ciclo biológico de prácticamente todas las especies que forman parte de l colección (casi 30). En efecto, el acuario de Valencia ha sido pionero en cerrar el ciclo y exhibir algunas especies a nivel nacional. Este es el caso de Tima formosa, Catostylus mosaicus, Chrysaora plocamia, Chrysaora lactea y Stomolophus meleagris, entre otras. El ICM-CSIC cuenta con una línea de investigación consolidada sobre medusas desde hace más de 20 años. La variabilidad de las condiciones ambientales, la estacionalidad de la mayoría de especies de medusas y la falta de conocimiento sobre sus ciclos biológicos, no garantizan su disponibilidad en la naturaleza. Bajo estas premisas, muchos de los conocimientos pluridisciplinares adquiridos a lo largo del tiempo son debidos a la posibilidad de trabajar en la Zona de Acuarios Experimentales (ZAE) del ICM-CSIC, un área destinada al manejo y cultivo de medusas en condiciones de cautividad permanente. Recientemente, y gracias a la amplia experiencia en cultivos de medusas, el equipo científico ha descrito, por primera vez, el cnidoma (conjunto de células urticantes, distribución y abundancia durante cada etapa del ciclo de vida) de la especie Pelagia noctiluca, la medusa más importante del mar Mediterráneo en sentido de abundancia y gravedad de su picadura. Trabajar bajo condiciones de cautividad, a su vez, ha permitido desarrollar proyectos de carácter industrial donde la disponibilidad de medusas es esencial para la optimización de los resultados. Gracias a la nueva colaboración entre el ICM-CSIC y el Oceanogràfic de Valencia se inicia una nueva era de transferencia de conocimientos entre ambas instituciones fomentando el intercambio de personal, actividades conjuntas y acciones de colaboración, en distintos campos relacionados con medusasPeer reviewe

Identifying biomarkers of pollutant exposure in ocean sentinels: Characterisation and optimisation of B-esterases in plasma from loggerhead turtles undergoing rehabilitation

9 pages, 4 figures, 4 tables, supplementary data https://doi.org/10.1016/j.chemosphere.2023.140770.-- Data will be made available on requestSea turtles are frequently proposed as indicator species for assessing ocean health. To faciliate the use of these species as bioindicators requires the development of tools for rapidly and effectively assessing individual health. Here, we collected 104 blood samples from 69 loggerhead sea turtles, Caretta caretta, undergoing rehabilitation to determine the connection between health status, the activity of B-esterases, and other biochemical parameters. To determine the optimal assay protocol for B-esterases, we measured the activity and kinetics of cholinesterases-(ChEs) and carboxylesterases (CEs) using 3 and 5 commercial substrates, respectively, at different assay conditions. IC50 values for the activity of B-esterases were calculated within a concentration range for model pesticide inhibitors. Turtles’ health status was determined via routine veterinary procedures. During rehabilitation (which was associated with improving health status), we observed a decrease in the activity of most enzymes (especially in acetylcholinesterase) alongside an increase in CE when using p-nitrophenyl acetate as a substrate. As such, it is possible that the activity rates of plasmatic B-esterases could serve as an indicator of health status. There is also high potential that B-esterases could be specifically sensitive to marine pollutants although to further validate this would require future studies to specifically correlate B-esterarse activities to pollutant concentrations in blood or excretaTo the institutional support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX 2019-000928-S). This work has received funding from CSIC Interdisciplinary Thematic Platform (PTI+) Interdisciplinary Platform for Sustainable Plastics towards a Circular Economy+ (PTI-SusPlast+)Peer reviewe

Two cases of pseudohermaphroditism in loggerhead sea turtles Caretta caretta

Two juvenile (curved carapace lengths: 28 and 30 cm) loggerhead sea turtles Caretta caretta with precocious male external characteristics were admitted to the ARCA del Mar rescue area at the Oceanogràfic Aquarium in Valencia, Spain, in 2009 and 2010. Routine internal laparoscopic examination and subsequent histopathology confirmed the presence of apparently healthy internal female gonads in both animals. Extensive tissue biopsy and hormone induction assays were consistent with female sex. To the best of our knowledge, this is the first report of pseudohermaphroditism in loggerhead sea turtles based on sexual external characteristics and internal laparoscopic examination. Our findings suggest that the practice of using external phenotypical characteristics as the basis for gender identification in sea turtles should be reevaluated. Future research should focus on detecting more animals with sexual defects and their possible effects on the sea turtle populationPeer reviewe

Brushes and Shelters as Low-Cost Environmental Enrichment Devices for Loggerhead Turtles (Caretta caretta) During Rehabilitation

7 pages, 3 figures, 3 tablesEnvironmental enrichment (EE) can significantly improve the welfare and health of captive animals. Despite the known benefits of EE, there has been a lack of research on the effects of EE devices (EED) for wild animals. Here, we quantified the time loggerhead turtles (Caretta caretta) in a rehabilitation center spent interacting with EEDs, specifically brushes and sheltering objects. We found turtles spent up to 50% of their time interacting with EEDs and demonstrated a preference to rest under sheltering objects (assisted resting), when compared with the open. Based on these findings, we propose the inclusion of brushes and sheltering objects as low-cost EEDs for improving the welfare of sea turtles in captivity.J.H.C. and D.M. acknowledge support from the GenT program of the Generalitat Valenciana (CIDEGENT/2021/058). This work was also supported by the Doctoral Career Development Fund awarded via the East of Scotland Bioscience Doctoral Training Partnership (EASTBIO) and funded by the UK Research and Innovation (UKRI) Biotechnology and Biological Sciences Research Council grant number BB/M010996/1. This research was funded under Fundación Oceanogràfic research grant OCE-32-22.With the institutional support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S)Peer reviewe

New insights into risk variables associated with gas embolism in loggerhead sea turtles (Caretta caretta) caught in trawls and gillnets

9 pages, 3 figures, 2 tables.-- Data Availability: The Statistical Appendix, R-code and data sets used in this study are freely available at the following link: http://osf.io/5t9qgTissue and blood gas embolism (GE) associated with fisheries bycatch are likely a widespread, yet underestimated, cause of sea turtle mortality. Here, we evaluated risk factors associated with tissue and blood GE in loggerhead turtles caught incidentally by trawl and gillnet fisheries on the Valencian coastline of Spain. Of 413 turtles (303 caught by trawl, 110 by gillnet fisheries), 54% (n = 222) exhibited GE. For sea turtles caught in trawls, the probability and severity of GE increased with trawl depth and turtle body mass. In addition, trawl depth and the GE score together explained the probability of mortality (P[mortality]) following recompression therapy. Specifically, a turtle with a GE score of 3 caught in a trawl deployed at 110 m had a P[mortality] of ~50%. For turtles caught in gillnets, no risk variables were significantly correlated with either the P[GE] or GE score. However, gillnet depth or GE score, separately, explained P[mortality], and a turtle caught at 45 m or with a GE score between 3 and 4 had a P[mortality] of 50%. Differences in the fishery characteristics precluded direct comparison of GE risk and mortality between these gear types. Although P[mortality] is expected to be significantly higher in untreated turtles released at sea, our findings can improve estimates of sea turtle mortality associated with trawls and gillnets, and help guide associate conservation effortsFunding for this project was provided by the Fundación Oceanografic and through a contract from the National Oceanic and Atmospheric AdministrationWith the institutional support of the ‘Severo Ochoa Centre of Excellence’ accreditation (CEX2019-000928-S)Peer reviewe