New species of Echinobothrium van Beneden, 1849 (Cestoda: Diphyllidea) from Indo-Pacific maskrays (Neotrygon Castelnau) off Australia and Borneo.

Four new diphyllidean species of the genus Echinobothrium van Beneden, 1849 are described from Indo-Pacific maskrays (Neotrygon Castelnau, Dasyatidae). Echinobothrium giraffaeous sp. n. from Neotrygon leylandi (Last) off northern Australia, Echinobothrium ivanovae sp. n. from Neotrygon orientalis Last, White et Serét off Borneo, and Echinobothrium bethae sp. n. from Neotrygon varidens (Garman) off Borneo are distinguished from all but one of the 33 valid species of the genus by the possession of the outermost A hooks with an extended base into which the bases of the three outermost B hooks are inserted. Echinobothrium rhynchobati (Khalil et Abdul-Salam, 1989) is the only known species with this unique feature, but its rostellum has a system of interlocking knobs and sockets that articulate bases of the A and B type hooks with one another, which is not present in any of the newly described species. Echinobothrium tyleri sp. n. from Neotrygon australiae Last, White et Serét off northern Australia is distinguished from all known species of Echinobothrium by its unique rostellar hook formula {2-3 18/17 2-3}. With the present addition of four new species, the central Indo-Pacific realm becomes the major hotspot for Echinobothrium, from which 13 species have been reported.

Fishing for oil and meat drives irreversible defaunation of deepwater sharks and rays.

The deep ocean is the last natural biodiversity refuge from the reach of human activities. Deepwater sharks and rays are among the most sensitive marine vertebrates to overexploitation. One-third of threatened deepwater sharks are targeted, and half the species targeted for the international liver-oil trade are threatened with extinction. Steep population declines cannot be easily reversed owing to long generation lengths, low recovery potentials, and the near absence of management. Depth and spatial limits to fishing activity could improve conservation when implemented alongside catch regulations, bycatch mitigation, and international trade regulation. Deepwater sharks and rays require immediate trade and fishing regulations to prevent irreversible defaunation and promote recovery of this threatened megafauna group.

Do electromagnetic fields from subsea power cables effect benthic elasmobranch behaviour? A risk-based approach for the Dutch Continental Shelf.

Subsea power cables cause electromagnetic fields (EMFs) into the marine environment. Elasmobranchs (rays, skates, sharks) are particularly sensitive to EMFs as they use electromagnetic-receptive sensory systems for orientation, navigation, and locating conspecifics or buried prey. Cables may intersect with egg laying sites, mating, pupping, and nursery grounds, foraging habitat and migration routes of elasmobranchs and the effects of encountering EMFs on species of elasmobranchs are largely unknown. Demonstrated behavioural effects are attraction, disturbance and indifference, depending on EMF characteristics, exposed life stage, exposure level and duration. We estimated exposure levels of elasmobranchs to subsea power cable EMFs, based on modelled magnetic fields in the Dutch Continental Shelf and compared these to reported elasmobranch sensory sensitivity ranges and experimental effect levels. We conclude that the risk from subsea power cables has a large uncertainty and varies per life stage and species ecology. Based on estimated no-observed effect levels (from 10-3 to 10-1 µT) we discuss what will probably be the most affected species and life stage for six common benthic elasmobranchs in the Southern North Sea. We then identify critical knowledge gaps for reducing the uncertainty in the risk assessments for EMFs effects on benthic elasmobranchs.

INTRAOCULAR PRESSURES OF AQUARIUM-HOUSED COWNOSE RAYS (RHINOPTERA BONASUS) WITH NORMAL AND ABNORMAL OPHTHALMIC EXAMS.

Cownose rays (Rhinoptera bonasus) are common elasmobranchs in zoos and aquaria; however, there is a lack of published information regarding ocular findings in this species. Intraocular pressure (IOP) was measured in a total of 52 cownose rays (Rhinoptera bonasus) from two unrelated aquaria (n = 22 from A1, n = 30 from A2) using a TonoVet rebound tonometer on two settings (dog = D, and unidentified species = P) as part of a full ophthalmologic examination. Adult (n = 38) and juvenile (n = 14) rays were sampled out of water briefly in sternal recumbency. Intraocular pressure (mean ± SD [range]) in the D setting (9.10 ± 2.57 [4-18] mmHg) was higher than the P setting (5.21 ± 2.32 [0-12] mmHg) (P<0.001). Statistical analysis revealed no difference in IOP between right and left eyes, and no correlation between body weight and IOP. No differences in IOP between sex, age group, and location were identified in either setting. However, a significant difference was observed between levels of severity of corneal disease in IOP D setting (P=0.006) and P setting (P=0.024), and levels of severity of intraocular disease in IOP D setting (P=0.034) only. This study provides baseline IOP values using rebound tonometry in aquarium-housed cownose rays with apparent corneal and intraocular lesions and reveals that the D setting may be more sensitive in identifying IOP changes in eyes with intraocular disease.

Levels of Total Mercury and Health Risk Assessment of Consuming Freshwater Stingrays (Chondrichthyes: Potamotrygoninae) of the Brazilian Amazon.

Mercury is an element with potential risk to fish and those who consume it. Thus, this study aimed to determine the levels of total mercury (THg), carry out a health risk assessment related to the consumption of the freshwater stingrays Potamotrygon motoro, and determine the physical and chemical properties of the water where stingrays occur. Stingrays of the species P. motoro were obtained from the Amazon River, and samples of the animals' musculature were collected to determine THg levels. Risk assessment was conducted using pre-established formulas of estimated monthly intake (EMI), maximum monthly intake rate (IRmm), and hazard quotient (HQ). Three population scenarios were evaluated, considering both sexes and differences between rural and urban areas. There was no relationship between weight and THg concentration nor between total length and THg concentration. Higher EMI values were observed in rural children; for the IRmm, male children had the lowest consumption levels. For the hazard quotient, there was a similarity between the three age groups when comparing the male and female sexes. In addition, the representatives of the rural area always had lower values than the urban area. Freshwater stingrays, like other elasmobranchs, can be crucial animal species because they act as sentinels in studies that assess harmful chemicals like mercury.

Evolution of the Critically Endangered Green Sawfish Pristis zijsron (Rhinopristiformes, Pristidae), Inferred from the Whole Mitochondrial Genome.

The green sawfish Pristis zijsron (Bleeker, 1851), a species of sawfish in the family Pristidae (Rhinopristiformes), mainly inhabits the Indo-West Pacific region. In this study, the complete mitochondrial genome of the critically endangered green sawfish is first described. The length of the genome is 16,804 bp, with a nucleotide composition of 32.0% A, 24.8% C, 13.1% G, and 30.0% T. It contains 37 genes in the typical gene order of fish. Two start (GTG and ATG) and two stop (TAG and TAA/T-) codons are found in the thirteen protein-coding genes. The 22 tRNA genes range from 67 bp (tRNA-Ser) to 75 bp (tRNA-Leu). The ratio of nonsynonymous substitution (Ka) and synonymous substitution (Ks) indicates that the family Pristidae are suffering a purifying selection. The reconstruction of Bayesian inference and the maximum likelihood phylogenetic tree show the same topological structure, and the family Pristidae is a monophyletic group with strong posterior probability. Pristis zijsron and P. pectinata form a sister group in the terminal clade. And the divergence time of Rhinopristiformes show that P. zijsron and P. pectinata diverged as two separate species in about Paleogene 31.53 Mya. Complete mitochondrial genomes of all five sawfishes have been published and phylogenetic relationships have been analyzed. The results of our study will provide base molecular information for subsequent research (e.g., distribution, conservation, phylogenetics, etc.) on this endangered group.

Exploring indicators of success in chronic total occlusion percutaneous coronary intervention with Stingray system-based antegrade dissection re-entry: Insights from retrospective analysis.

BACKGROUND: The predictors of success of chronic total occlusion (CTO) percutaneous coronary intervention (PCI) through antegrade dissection and re-entry (ADR) using the Stingray system (Stingray ADR) remain elusive, mainly owing to the lack of consecutive angiographic and procedural records of patients. OBJECTIVES: This study aimed to identify indicators that can determine the success of CTO PCI performed using the Stingray ADR technique. METHODS: The clinical data of 115 patients who underwent CTO PCI through Stingray ADR at the same cardiac center were retrospectively and consecutively collected. Multivariate logistic regression analysis was performed to investigate the indicators of the success of ADR attempts. RESULTS: The technical success rate of Stingray ADR in CTO PCI was 72.2%. The overall technical success rate of CTO recanalization was 78.3% in all CTO PCIs having used Stingray Low Profile balloon. Vessel calcification (odds ratio [OR]: 4.03; 95% confidence interval [CI]: 1.49-11.88; p = 0.008), and retrograde puncture indicator (OR: 4.89; 95% CI: 1.51-17.11; p = 0.009) were identified as independent positive predictors. Blunt/no stump proximal to the occlusion segment (OR: 0.22; 95% CI: 0.06-0.64; p = 0.009), decision time before Stingray ADR (per 1 h increase) (OR: 0.54; 95% CI: 0.31-0.92; p = 0.026), operation duration of Stingray ADR (per 10 min increase) (OR: 0.62; 95% CI: 0.40-0.94; p = 0.028), and puncture site at the intraplaque region (OR: 0.24; 95% CI: 0.06-0.84; p = 0.026) were identified as the four negative independent predictors. CONCLUSIONS: This study revealed independent predictors of the success of CTO PCI performed using the Stingray ADR technique. As for CTO characteristics, the presence of calcification in the CTO segment and a tapered stump proximal to the lesion site can facilitate successful Stingray ADR. As for the procedures, the success rate of Stingray ADR can be improved by initiating the technique decisively and promptly, operating the system quickly and accurately and creating a puncture in the distal cap region of CTO under retrograde guidance.

Accuracy of Emergency Medicine Residents Using Point-of-Care Ultrasound (POCUS) to Detect Retained Stingray Barbs.

BACKGROUND: Stingray envenomation is a common presenting complaint for coastal emergency departments in the United States. Currently, radiograph is the gold standard to evaluate for a retained stingray barb, but ultrasound may be a useful tool to detect retained barbs. OBJECTIVE: To determine if emergency medicine residents could use ultrasound to identify stingray barbs embedded in animal tissue models. A secondary objective was to determine if resident experience affected their ability to detect stingray barbs. METHODS: Thirty-two emergency medicine residents participated in the study. After a short didactic session on foreign body identification with ultrasound, they rotated through six simulation stations and were asked to identify whether a stingray barb was present in pig and chicken tissue models. They were given 2 min per model to identify the presence, size, and depth of a stingray barb. Pre- and postexperiment surveys were collected to assess the residents' level of experience and confidence regarding foreign body identification using ultrasound. RESULTS: Residents accurately identified barbs in chicken drumsticks with a sensitivity of 72.92% (95% confidence interval [CI] 63.89-81.48) and a specificity of 64.58% (95% CI 54.16-74.08), and in pig's feet with a sensitivity of 50.00% (95% CI 39.62-60.38) and specificity of 68.75% (95% CI 58.48-77.82). There was no statistically significant difference regarding accuracy for any outcome measured based on experience or level of training. CONCLUSIONS: The use of point-of-care ultrasound by novice sonographers lacks sensitivity to identify retained stingray barbs in animal models and is not significantly impacted by resident experience with point-of-care ultrasound.

An integrated system to assess marine extinctions.

More than 20 global marine extinctions and over 700 local extinctions have reportedly occurred during the past 500 years. However, available methods to determine how many of these species can be confidently declared true disappearances tend to be data-demanding, time-consuming, and not applicable to all taxonomic groups or scales of marine extinctions (global [G] and local [L]). We developed an integrated system to assess marine extinctions (ISAME) that can be applied to any taxonomic group at any geographic scale. We applied the ISAME method to 10 case studies to illustrate the possible ways in which the extinction status of marine species can be categorized as unverified, possibly extinct, or extinct. Of the 10 case studies we assessed, the ISAME method concludes that 6 should be categorized as unverified extinctions due to problems with species' identity and lack of reliable evidence supporting their disappearance (periwinkle-Littoraria flammea [G], houting-Coregonus oxyrinchus [G], long-spined urchin-Diadema antillarum [L], smalltooth sawfish-Pristis pectinata [L], and largetooth sawfish-P. pristis [L]). In contrast, ISAME classified the Guadalupe storm-petrel (Oceanodroma macrodactyla [G]) and the lost shark (Carcharhinus obsolerus [G]) as possibly extinct because the available evidence indicates that their extinction is plausible-while the largetooth sawfish [L] and Steller's sea cow (Hydrodamalis gigas [G]) were confirmed to be extinct. Determining whether a marine population or species is actually extinct or still extant is needed to guide conservation efforts and prevent further biodiversity losses.

Morphological and molecular characterization of Chloromyxum dasyatidis n. sp. (Myxosporea: Chloromyxidae) in the common stingray Dasyatis pastinaca (Linnaeus) from Tunisian waters (Central Mediterranean Sea).

A new species of Chloromyxum Mingazzini, 1890, C. dasyatidis n. sp., is described from the gallbladder of the elasmobranch fish Dasyatis pastinaca (Linnaeus) from the Mediterranean coast off Tunisia. Mature myxospores were subspherical measuring 13.0 ± 0.3 (12.5-13.9) µm in length, 11.3 ± 0.5 (10.2-12.2) µm in width, and 11.8 ± 0.5 (11.4-12.7) µm in thickness. Four pyriform polar capsules 4.4 ± 0.3 (4.0-4.9) µm long and 3.5 ± 0.3 (3.0-3.8) µm wide. Valves were ornamented by 5 to 7 elevated surface ridges and displayed short caudal filaments. Chloromyxum dasyatidis n. sp. had an overall prevalence of infection of 38.5%, with significant seasonal variation, being more prevalent during summer (60.0%) and autumn (70.8%). Phylogenetically, this species grouped within the Chloromyxum sensu stricto lineage, forming a small clade together with the genetically more similar species C. acuminatum and C. myolibati, both from stingray hosts.

Progesterone receptors in extratesticular ducts of the Amazonian stingray Potamotrygon wallacei: A potential role in sperm maturation and aggregate formation.

In cururu stingray (Potamotrygon wallacei Carvalho, Rosa and Araújo 2016) males, plasma progesterone (P4) levels appear to be associated with spermiation events. However, the specific contribution of P4 in sperm maturation via extratesticular ducts in this stingray species is unknown. With the aim of filling this knowledge gap, this study examined the morphology and the presence of progesterone receptors (PR) in the ducts, and analyzed the relationship of progesterone (P4) with sperm maturation and formation of aggregates. Morphological analysis showed that a columnar pseudostratified epithelium with stereocilia lined all the attached ducts. In active males, the secretory cells of the epididymis and the Leydig glands presented PR; however, these receptors were not found in the distal region of the epididymis (essential for nurturing and capacitation events) of regressing males. In the seminal vesicles of active males, the spermatozoa are parallelly aligned and embedded in a matrix to form the spermatozeugmata. The matrixes are formed by proteins secreted by the ducts and Sertoli cell cytoplasts. These structures presented PR, which suggests that P4 engages in sperm metabolism during storage. Our findings allude to the potential role of P4 in regulating the development and function of the attached ducts in different reproductive phases. Furthermore, P4 seems to be an essential component for regulating sperm progress, protein secretion, aggregate formation, and maintenance of sperm during storage in this freshwater stingray.

Genomic Characteristics of Okamejei kenojei and the Implications to Its Evolutionary Biology Study.

Okamejei kenojei is an important economic species widely distributed in shallow coastal waters of the western North Pacific. In this study, the whole-genome survey analysis of O. kenojei was conducted to reveal its genomic characteristics. The genome size was estimated to be 2027.44 Mb, the repeat sequence content was 44.90%, and the heterozygous ratio was 1.04%. The mitochondrial genome excavated from the sequencing data was 16,974 bp, and it can form the closed circular molecule. The phylogenetic tree based on 13 protein-coding gene sequences supported the validity of Okamejei and assisted the conclusion that Raja porosa was the junior synonym of O. kenojei. Plenty of potential microsatellite loci were identified, and the distribution frequency was estimated to be approximately 236.3 SSRs per Mb. Among all motif types of microsatellites, the dinucleotide repeats were dominant (82.59%), followed by the trinucleotide repeats (8.05%), tetranucleotide repeats (5.80%), pentanucleotide repeats (2.83%), and hexanucleotide repeats (0.72%). The results of the present study could not only provide useful information for understanding the genome structure and functional characteristics of O. kenojei, but also lay the foundation for the subsequent mapping of the whole genome.

Elemental analysis of vertebrae discerns diadromous movements of threatened non-marine elasmobranchs.

River sharks (Glyphis spp.) and some sawfishes (Pristidae) inhabit riverine environments, although their long-term habitat use patterns are poorly known. We investigated the diadromous movements of the northern river shark (Glyphis garricki), speartooth shark (Glyphis glyphis), narrow sawfish (Anoxypristis cuspidata), and largetooth sawfish (Pristis pristis) using in situ laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) on vertebrae to recover elemental ratios over each individual's lifetime. We also measured elemental ratios for the bull shark (Carcharhinus leucas) and a range of inshore and offshore stenohaline marine species to assist in interpretation of results. Barium (Ba) was found to be an effective indicator of freshwater use, whereas lithium (Li) and strontium (Sr) were effective indicators of marine water use. The relationships between Ba and Li and Ba and Sr were negatively correlated, whereas the relationship between Li and Sr was positively correlated. Both river shark species had elemental signatures indicative of prolonged use of upper-estuarine environments, whereas adults appear to mainly use lower-estuarine environments rather than marine environments. Decreases in Li:Ba and Sr:Ba at the end of the prenatal growth zone of P. pristis samples indicated that parturition likely occurs in fresh water. There was limited evidence of prolonged riverine habitat use for A. cuspidata. The results of this study support elemental-environment relationships observed in teleost otoliths and indicate that in situ LA-ICP-MS elemental characterization is applicable to a wide range of elasmobranch species as a discriminator for use and movement across salinity gradients. A greater understanding of processes that lead to element incorporation in vertebrae, and relative concentrations in vertebrae with respect to the ambient environment, will improve the applicability of elemental analysis to understand movements across the life history of elasmobranchs into the future.

Identification and characterization of a polyomavirus in the thornback skate (Raja clavata).

Members of the family Polyomaviridae have a circular double-stranded DNA genome that have been identified in various hosts ranging from mammals to arachnids. Here we report the identification and analysis of a complete genome sequence of a novel polyomavirus, Raja clavata polyomavirus (RcPyV1), from a cartilaginous fish, the thornback skate (Raja clavata). The genome sequence was determined using a metagenomics approach with an aim to provide baseline viral data in cartilaginous fish in different ecosystems. The RcPyV1 genome (4,195 nucleotides) had typical organization of polyomavirus, including early antigens (small T; Large T) encoded on one strand and late viral proteins (VP1; VP2) on the complementary strand. Maximum-likelihood phylogenetic analysis of the large T-antigen revealed that RcPyV1 clusters with a polyomavirus obtained from another cartilaginous fish, the guitarfish polyomavirus 1 (GfPyV1). These two share ~ 56% pairwise identity in LT and VP1 protein sequences. These analyses support the hypothesis that cartilaginous fishes have a specific lineage of polyomaviruses.

Okamejei picta sp. nov., a new rajid skate from the South China Sea (Rajiformes: Rajidae).

A new species of Okamejei is described based on two adult males collected from deep waters in the South China Sea. The new species, Okamejei picta sp. nov., is readily distinguished from most other congeners in having densely scattered black spots on dorsal disc. Okamejei hollandi and O. mengae is quite similar to the new species by their spot patterns on dorsal disc, but the new species differs from the former by a combination of characters: a yellowish brown dorsal surface densely covered with small, circular to irregular-shaped black spots; blotches on dorsal disc indistinct; posterior ocellus absent; ventral disc white; disc length 45.0-47.7% TL; distance between cloaca to caudal-fin tip 53.6-55.1% TL; trunk centra 31; total basal radials 73-76, morphology of clasper terminal skeleton, and lacking component funnel at the clasper end.

Ontogenetic morphometry of the brown smoothhound shark Mustelus henlei with implications for ecology and evolution.

The central tenet of ecomorphology links ecological and morphological variation through the process of selection. Traditionally used to rationalise morphological differences between taxa, an ecomorphological approach is increasingly being utilised to study morphological differences expressed through ontogeny. Elasmobranchii (sharks, rays and skates) is one clade in which such ontogenetic shifts in body form have been reported. Such studies are limited to a relatively small proportion of total elasmobranch ecological and morphological diversity, and questions remain regarding the extent to which ecological selection are driving observed morphometric trends. In this study, we report ontogenetic growth trajectories obtained via traditional linear morphometrics from a large data set of the brown smoothhound shark (Mustelus henlei). We consider various morphological structures including the caudal, dorsal and pectoral fins, as well as several girth measurements. We use an ecomorphological approach to infer the broad ecological characteristics of this population and refine understanding of the selective forces underlying the evolution of specific morphological structures. We suggest that observed scaling trends in M. henlei are inconsistent with migratory behaviour, but do not contradict a putative trophic niche shift. We also highlight the role of predation pressure and sex-based ecological differences in driving observed trends in morphometry, a factor which has previously been neglected when considering the evolution of body form in sharks.

A New Species of Branchellion (Hirudinea: Piscicolidae) Parasitizing the Gills of Short-Tail Stingrays (Batoidea: Dasyatidae) from the West Pacific.

A new fish leech, Branchellion brevicaudatae sp. n., is described based on specimens parasitizing the gills of the short-tail stingray, Bathytoshia brevicaudata (Hutton, 1875), collected from Japanese waters. The new species can be distinguished from other congeners by having: i) pulsating vesicles emerging from posterior base of branchiae, one pair per somite; ii) dorsal white spots, not arranged in longitudinal row; and iii) blackish body. A phylogenetic tree based on partial sequences of the mitochondrial cytochrome c oxidase subunit I gene from the new species and other piscicolid worms showed that the new species is sister to Branchellion torpedinis Savigny, 1822. This is the first record of Branchellion Savigny, 1822 from Japanese waters.

Characterization of Some Dermato-Cosmetic Preparations with Marine Lipids from Black Sea Wild Stingray.

The traditional knowledge about the therapeutic and nutritional value of fish has been unanimously recognized among the population since ancient times. So, thanks to the therapeutic virtues of these marine animals, it was possible to develop therapies for certain pathologies as well as the use of bioactive compounds as adjunctive therapies incorporated into the treatment regimen of patients. In the present study, stingray liver oil from wild species collected from the Romanian coast of the Black Sea was isolated and analyzed. Fatty acid analysis was performed by gas chromatography. The analysis of the distribution of fatty acids in the composition of stingray liver oil indicates a ratio of 2.83 of omega 3 fatty acids to omega 6, a ratio of 1.33 of polyunsaturated fatty acids to monounsaturated fatty acids, an iodine index of 111.85, and a total percentage of 68.98% of unsaturated fatty acids. Stingray liver oil was used to evaluate the healing action after preparing a fatty ointment. According to the experimental data, a complete regeneration capacity of the wounds was noted in 12 days without visible signs. Four emulgels with stingray liver oil were formulated and analyzed from a rheological and structural point of view in order to select the optimal composition, after which the anti-inflammatory effect on inflammation caused in laboratory rats was studied and an anti-inflammatory effect was found significant (a maximum inhibitory effect of 66.47% on the edemas induced by the 10% kaolin suspension and 65.64% on the edemas induced by the 6% dextran solution).