First molecular determination of herpesvirus from two mysticete species stranded in the Mediterranean Sea

BACKGROUND Herpesvirus can infect a wide range of animal species: mammals, birds, reptiles, fish, amphibians and bivalves. In marine mammals, several alpha- and gammaherpesvirus have been identified in some cetaceans and pinnipeds species. To date, however, this virus has not been detected in any member of the Balaenoptera genus. CASE PRESENTATION Herpesvirus was determined by molecular methods in tissue samples from a male fin whale juvenile (Balaenoptera physalus) and a female common minke whale calf (Balaenoptera acutorostrata) stranded on the Mediterranean coast of the Region of Valencia (Spain). Samples of skin and penile mucosa from the fin whale and samples of skin, muscle and central nervous system tissue from the common minke whale tested positive for herpesvirus based on sequences of the DNA polymerase gene. Sequences from fin whale were identical and belonged to the Alphaherpesvirinae subfamily. Only members of the Gammaherpesvirinae subfamily were amplified from the common minke whale, and sequences from the muscle and central nervous system were identical. Sequences in GenBank most closely related to these novel sequences were viruses isolated from other cetacean species, consistent with previous observations that herpesviruses show similar phylogenetic branching as their hosts. CONCLUSIONS To our knowledge, this is the first molecular determination of herpesvirus in the Balaenoptera genus. It shows that herpesvirus should be included in virological evaluation of these animals

New insights into the pathogenesis and transmission of Brucella pinnipedialis: systemic infection in two bottlenose dolphins ( Tursiops truncatus)

The emergence of Brucella infections in marine mammals is a growing concern. The present study reports two cases of systemic Brucella pinnipedialis infection detected in bottlenose dolphins (Tursiops truncatus) pair stranded together in the Cantabrian coast of Spain. Both animals showed systemic lesions associated with the Brucella infection, more severe in the younger dolphin, considered the likely offspring of the other individual. Real-time PCR, bacterial culture, and whole-genome sequencing were used to detect and characterize the Brucella strains involved in both dolphins. The phylogenetic analysis performed on the Brucella genomes retrieved revealed that the species involved was B. pinnipedialis (ST25). Both animals resulted seropositive in a commercial multispecies blocking ELISA but tested negative in the standard Rose Bengal test. To the best of our knowledge, this is the first report of a systemic infection resulting in various lesions associated with Brucella pinnipedialis (ST25) in two bottlenose dolphins. It is also the initial isolation of Brucella in the milk of a non-pregnant or non-aborting female cetacean likely stranded with its offspring. These findings provide new insights into the epidemiology and clinical impact of B. pinnipedialis infection in cetaceans and underscore the importance of continued diagnostic surveillance to gain better understanding of brucellosis effects and transmission in marine mammal populations.Este trabajo ha contado con el apoyo de un convenio de colaboración entre la Fundación Oceanogràfic (Valencia), la Consejería de Medio Rural, Ganadería, Pesca, Alimentación y Medio Ambiente del Gobierno de Cantabria y el Centro de Vigilancia Sanitaria Veterinaria (VISAVET) de la Universidad Complutense de Madrid. Este convenio permite la recogida de muestras de cadáveres de cetáceos varados y aporta fondos para la realización de estudios post-mortem. Ignacio Vargas-Castro es beneficiario de una beca FPU del Ministerio de Ciencia, Innovación y Universidades. El trabajo del CITA fue apoyado por el Gobierno de Aragón (Grupo de Investigación A21_23R). André E. Moura recibió el apoyo del Centro Nacional de Ciencia de Polonia (beca de investigación Sonata 2018/31/D/NZ8/02835) y de la Agencia Nacional Polaca de Intercambio Académico (programa NAWA Ulam PPN/ ULM/2019/1/00162).milkBrucella pinnipedialisbottlenose dolphinmarine mammalssystemic infectionhealth surveillancetransmissionwhole-genome sequencingPublishe

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

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

Effects of 3 Tesla magnetic resonance imaging exposure on the behavior and orientation of homing pigeons <i>Columba livia domestica</i>

Homing pigeons (Columba livia domestica) were used to test whether clinical magnetic resonance (MR) imaging disrupts orientation of animals that sense the earth’s magnetic field. Thirty young pigeons were randomly separated into three groups (n = 10/group). Two groups were anaesthetized and exposed to either a constant (no sequence) or a varying (gradient echo and echo planar sequences) magnetic field within a 3 Tesla MR unit for 15 minutes. The control group was not exposed to the MR field but shared all other aspects of the procedure. One day later, animals were released from a site they had never visited, 15 km from the home loft. Three weeks after the procedure, animals were released from a different unfamiliar site 30 km from the loft. Measured variables included the time to disappear from sight (seconds), vanishing bearing (angle), and the time interval from release to entering the home loft (hours). On first release, the group exposed to varying field gradients during image acquisition using 2 different standard sequences showed more variability in the vanishing bearing compared to the other groups (p = 0.0003 compared to control group), suggesting interference with orientation. Other measures did not show significant differences between groups. On second release, there were no significant differences between groups. Our results on homing pigeons show that regular clinical MR imaging exposure may temporarily affect the orientation of species that have magnetoreception capabilities. If exposure to MR imaging disrupted processes that are not specific to magnetoreception, then it may affect other species and other capabilities as well.</p

First molecular determination of herpesvirus from two mysticete species stranded in the Mediterranean Sea

BACKGROUND Herpesvirus can infect a wide range of animal species: mammals, birds, reptiles, fish, amphibians and bivalves. In marine mammals, several alpha- and gammaherpesvirus have been identified in some cetaceans and pinnipeds species. To date, however, this virus has not been detected in any member of the Balaenoptera genus. CASE PRESENTATION Herpesvirus was determined by molecular methods in tissue samples from a male fin whale juvenile (Balaenoptera physalus) and a female common minke whale calf (Balaenoptera acutorostrata) stranded on the Mediterranean coast of the Region of Valencia (Spain). Samples of skin and penile mucosa from the fin whale and samples of skin, muscle and central nervous system tissue from the common minke whale tested positive for herpesvirus based on sequences of the DNA polymerase gene. Sequences from fin whale were identical and belonged to the Alphaherpesvirinae subfamily. Only members of the Gammaherpesvirinae subfamily were amplified from the common minke whale, and sequences from the muscle and central nervous system were identical. Sequences in GenBank most closely related to these novel sequences were viruses isolated from other cetacean species, consistent with previous observations that herpesviruses show similar phylogenetic branching as their hosts. CONCLUSIONS To our knowledge, this is the first molecular determination of herpesvirus in the Balaenoptera genus. It shows that herpesvirus should be included in virological evaluation of these animals.Centro de Vigilancia Sanitaria Veterinaria (VISAVET)TRUEpu

Effects of 3 Tesla magnetic resonance imaging exposure on the behavior and orientation of homing pigeons <i>Columba livia domestica</i>

Funding: PLT was funded by the US Office of Naval Research (https://www.onr.navy.mil) Grant Number N00014-18-1-2062 and N00014-20-1-2709. The employees of the Fundación Oceanogràfic de la Comunidad Valenciana were funded by the Fundación.Homing pigeons (Columba livia domestica) were used to test whether clinical magnetic resonance (MR) imaging disrupts orientation of animals that sense the earth’s magnetic field. Thirty young pigeons were randomly separated into three groups (n = 10/group). Two groups were anaesthetized and exposed to either a constant (no sequence) or a varying (gradient echo and echo planar sequences) magnetic field within a 3 Tesla MR unit for 15 minutes. The control group was not exposed to the MR field but shared all other aspects of the procedure. One day later, animals were released from a site they had never visited, 15 km from the home loft. Three weeks after the procedure, animals were released from a different unfamiliar site 30 km from the loft. Measured variables included the time to disappear from sight (seconds), vanishing bearing (angle), and the time interval from release to entering the home loft (hours). On first release, the group exposed to varying field gradients during image acquisition using 2 different standard sequences showed more variability in the vanishing bearing compared to the other groups (p = 0.0003 compared to control group), suggesting interference with orientation. Other measures did not show significant differences between groups. On second release, there were no significant differences between groups. Our results on homing pigeons show that regular clinical MR imaging exposure may temporarily affect the orientation of species that have magnetoreception capabilities. If exposure to MR imaging disrupted processes that are not specific to magnetoreception, then it may affect other species and other capabilities as well.Publisher PDFPeer reviewe

To Swim or Not to Swim: Potential Transmission of Balaenophilus manatorum (Copepoda: Harpacticoida) in Marine Turtles.

Species of Balaenophilus are the only harpacticoid copepods that exhibit a widespread, obligate association with vertebrates, i.e., B. unisetus with whales and B. manatorum with marine turtles and manatees. In the western Mediterranean, juveniles of the loggerhead sea turtle, Caretta caretta are the only available hosts for B. manatorum, which has been found occurring at high prevalence (>80%) on them. A key question is how these epibionts are transmitted from host to host. We investigated this issue based on experiments with live specimens of B. manatorum that were cultured with turtle skin. Specimens were obtained from head-started hatchlings of C. caretta from the western Mediterranean. Hatched nauplii crawled only on rough substrates and lacked the ability to swim. Only copepodites IV and V, and adults, were able to perform directional swimming. Legs 2, 3 and 4 played a major role in swimming and were only well-developed in these stages. Nauplii reared in wells with turtle skin readily fed on this item. Late copepodites and adults also fed on turtle skin but did not consume other potential food items such as fish skin, baleen plates or planktonic algae. Evidences suggest that the transmission of B. manatorum should rely on hosts' bodily contacts and/or swimming of late developmental stages between spatially close hosts. The possibility of long-ranged dispersal is unlikely for two reasons. First, all developmental stages seem to depend on turtle skin as a food resource. Second, the average clutch size of ovigerous females was small (< 70 eggs) for free-living phases to successfully contact turtles that occur at very low densities (< 0.6 turtles·km-2) in the western Mediterranean. The high prevalence of B. manatorum in loggerhead turtles in this area raises the question whether these turtles have contacts, or tend to closely aggregate, more than is currently believed