Enterolith with a bonefish spine nidus in a Brazilian cownose ray Rhinoptera brasiliensis (Müller & Henle) (Myliobatiformes: Rhinopteridae).

An adult female Brazilian cownose ray (Rhinoptera brasiliensis), housed in Ubatuba Aquarium died after loss of appetite period. During necropsy, an enterolith was discovered partially obstructing the intestinal lumen. Examination of the enterolith revealed a bonefish spine nidus. Enterolithiasis has been linked to multiple factors including diet, genetics, alkaline intestinal environments and, as in this particular case, the ingestion and retention of foreign bodies. The composition of this enterolith from a cownose ray was primarily monohydrocalcite. This short communication, apparently the first to report enterolithiasis in stingrays, emphasizes the need for post-mortem examinations of carcasses on macroscopic and microscopic levels.

Characterization of the complete mitochondrial genome of the brazilian cownose ray Rhinoptera brasiliensis (Myliobatiformes, Rhinopteridae) in the western Atlantic and its phylogenetic implications.

BACKGROUND: The Brazilian cownose ray, Rhinoptera brasiliensis has undergone a global population reduction and is currently classified by IUCN as Vulnerable. This species is sometimes confused with Rhinoptera bonasus, the only external diagnostic characteristic to distinguish between both species is the number of rows of tooth plates. Both cownose rays overlap geographically from Rio de Janeiro to the western North Atlantic. This calls for a more comprehensive phylogenetic assessment using mitochondria DNA genomes to better understand the relationships and delimitation of these two species. METHODS AND RESULTS: The mitochondrial genome sequences of R. brasiliensis was obtained by next-generation sequencing. The length of the mitochondrial genome was 17,759 bp containing 13 protein-coding genes (PCGs), two ribosomal RNA (rRNA) genes, 22 transfer RNA (tRNA) genes, and a non-coding control region (D-loop). Each PCG was initiated by an authoritative ATG codon, except for COX1 initiated by a GTG codon. Most of the PCGs were terminated by a complete codon (TAA/TAG), while an incomplete termination codon (TA/T) was found in five out of the 13 PCGs. The phylogenetic analysis showed that R. brasiliensis was closely related to R. steindachneri whereas the reported mitogenome as R. steindachneri (GenBank accession number KM364982), differs from multiple mitocondrial DNA sequences of R. steindachneri and is nearly identical to that of R. javanica. CONCLUSION: The new mitogenome determined in this study provides new insight into the phylogenetic relationships in Rhinoptera, while providing new molecular data that can be applied to population genetic studies.

Extending records of albinism and skin disorders in American cownose rays to southeastern Brazil.

Albinism is a genetic condition expressed as a lack of integumentary and retinal melanin. Although documented across many vertebrate species, albinism and other skin disorders are rarely observed in elasmobranchs (sharks and rays). The present study describes the first confirmed case of albinism in American cownose ray (Rhinoptera bonasus), and three other juveniles of the species with undetermined skin disorders observed in São Paulo, southeastern Brazil. Pigmentation disorders have already been observed in American cownose rays from the North Atlantic, including two cases of leucism and a possible albinism. Based on the results, it was discussed the possible consequences of albinism for the ray survival as well as the possible causes for the undetermined skin disorders.

Physiologic responses of Cownose Rays following freshwater dips for treatment of capsalid monogeneans.

OBJECTIVE: Two Cownose Rays Rhinoptera bonasus were presented for management of a severe capsalid monogenean infection Benedeniella posterocolpa in a mixed species habitat at an aquarium. METHODS: A series of freshwater (FW) dips were elected to mitigate parasite resistance to praziquantel due to the endemic monogenean population in the system. A pretreatment blood sample was opportunistically obtained, and subsequent samples were processed due to development of clinical signs. RESULT: While the first FW dip was tolerated well and did not induce abnormal behavior, the second led to one individual presenting with tachypnea, pallor, and lethargy. Marked hematological abnormalities requiring medical intervention occurred in both individuals after the second dip. After treatment with elasmobranch Ringer's fluid therapy, sodium bicarbonate, prednisolone acetate, and vitamin E/selenium in the more severely affected ray, and elasmobranch Ringer's alone in the second ray, hematologic derangements and symptoms resolved. Hemoconcentration, hyperproteinemia, hyperlactatemia, and hyperglycemia were attributed to a stress response. A selective loss of urea after exposure to FW resulted in decreased plasma osmolality. CONCLUSION: Given the severity of the stress response and associated complications observed, hyposalinity treatments should be utilized with caution in this species. If this therapy is employed, the clinician is advised to be prepared to administer supportive care coinciding with the FW dip if necessary.

Genetic evidence supports a range extension for the Brazilian cownose ray Rhinoptera brasiliensis in the western North Atlantic.

We report 24 new records of the Brazilian cownose ray Rhinoptera brasiliensis outside its accepted geographic range. Sequencing of a 442-base pair portion of the mitochondrial NADH dehydrogenase subunit 2 gene for 282 Rhinoptera samples revealed eight records off the east coast of the USA and 16 from the eastern Gulf of Mexico. Both sexes of all life stages were documented in all seasons over multiple years in the Indian River and Lake Worth lagoons, Florida, indicating that their range extends further in the western North Atlantic than previously described.

Variation in cownose ray Rhinoptera neglecta abundance and group size on the central east coast of Australia.

Here, we provide baseline information about the relative abundance and group size of the Australian cownose ray Rhinoptera neglecta on the central east coast of Australia. Using drone monitoring over 2 years, we completed 293 transects, each 2 km in length, at four locations distributed along c.100 km of coast. In total, 5979 R. neglecta were observed with overall relative abundance (±SE) of, 20.4 (±3.3) individuals per transect. The numbers of R. neglecta varied among locations, with the highest density found off the beach adjacent to the river mouth at Evans Head. The number of rays observed also decreased with increasing wind speed. While some of this relationship was probably associated with visibility, R. neglecta may also move offshore during strong winds. We found no evidence that R. neglecta was under significant threat. Additionally, our cost-effective surveys demonstrate the utility of aerial drones in fisheries conservation biology.

TEMPORAL STABILITY OF IN VITRO VENOUS BLOOD GAS, pH, AND LACTATE VALUES OF COWNOSE RAYS (RHINOPTERA BONASUS) AND RED-EARED SLIDER TURTLES (PSEUDEMYS SCRIPTA ELEGANS).

This study assessed the in vitro temporal changes that occur in blood pH and lactate concentrations for an elasmobranch species and a chelonian species, as well as blood gases (partial pressures of carbon dioxide [pCO2] and oxygen [pO2]) for a chelonian species, with a portable clinical point-of-care analyzer. Blood samples were collected from 10 cownose rays (Rhinoptera bonasus) and 10 red-eared sliders (Pseudemys scripta elegans), stored on ice, and serially analyzed at six time points up to 90 min postcollection. Results indicate that analysis should be conducted as soon as possible after blood collection for these species, with immediate analysis being preferred. However, if analysis must be delayed, syringes may be capped, placed on ice, and analyzed at a later time. Analysis within 90 min provided clinically acceptable results for pH and lactate in both species and for pCO2 in red-eared sliders, whereas substantial artifactual increases of pO2 were seen in red-eared sliders.

RADIOGRAPHIC DETERMINATION OF GASTRIC EMPTYING AND GASTROINTESTINAL TRANSIT TIME IN COWNOSE RAYS (RHINOPTERA BONASUS) AND WHITESPOTTED BAMBOO SHARKS (CHILOSCYLLIUM PLAGIOSUM) AND THE EFFECT OF METOCLOPRAMIDE ON GASTROINTESTINAL MOTILITY.

Gastrointestinal (GI) pathology is common in elasmobranchs; however, information regarding normal GI transit time and the effect of therapeutics on GI motility is lacking. The objective of this study was to determine baseline gastric emptying and GI transit times in cownose rays (Rhinoptera bonasus) and whitespotted bamboo sharks (Chiloscyllium plagiosum) via radiographic barium sulfate contrast studies. Additionally, a pilot study was undertaken to determine the effect of metoclopramide on GI transit time in whitespotted bamboo sharks. Eight cownose rays and eight whitespotted bamboo sharks were administered a 98% w/w barium sulfate suspension at 8 ml/kg via orogastric tube. Post-contrast radiographs were obtained at 2 min, 3, 6, 12, and 23 hr for rays; and 2 min, 3, 6, 9, 12, 16, 25, 30, 36, and every 12 hr until complete gastric emptying occurred for sharks. In cownose rays, the mean and standard error were established for time of initial spiral colon filling (3.4 ± 0.4 hr), complete spiral colon opacification (12 ± 0 hr), initial spiral colon emptying (21.6 ± 1.4 hr), and complete gastric emptying (23 ± 0 hr). In bamboo sharks, the mean and standard error were established for time of initial spiral colon filling (5.3 ± 0.5 hr), complete spiral colon opacification (12.4 ± 1.3 hr), initial spiral colon emptying (22.5 ± 2.7 hr), and complete gastric emptying (39.9 ± 3.3 hr). Cownose rays had a significantly shorter time to spiral colon filling and complete gastric emptying compared with bamboo sharks (P < 0.05). Whitespotted bamboo sharks (n = 8) were administered metoclopramide (0.4 mg/kg orally once daily for 10 days) and the barium series was repeated. Complete gastric emptying time was significantly shorter in treated sharks compared with control (P < 0.05), suggesting that metoclopramide may be a useful therapeutic for GI motility disorders in elasmobranchs.

REPRODUCTIVE HORMONE PATTERNS IN MALE AND FEMALE COWNOSE RAYS ( RHINOPTERA BONASUS) IN AN AQUARIUM SETTING AND CORRELATION TO ULTRASONOGRAPHIC STAGING.

Reproductive management of cownose rays ( Rhinoptera bonasus) under professional care plays an important role in conservation of the species, but hormone and ultrasonographic analyses of their 12-mo reproductive cycle have not been documented previously. Plasma reproductive hormone concentrations (17B-estradiol, progesterone, testosterone, and androstenedione) were measured monthly via radioimmunoassay for 1 yr in an aquarium-managed population of adult females ( n = 15) and males ( n = 5). Ultrasounds of the uterus were performed each month at the time of sample collection to identify gestation stage (0-5) based on a previously developed in-house staging system. Stages were correlated to hormone concentrations to track progression through pregnancy. Thirteen females were reproductively active, and each produced one pup in March-June, similar to timing for free-ranging populations. Female estradiol increased steadily throughout gestation from stages 0 to 5, while progesterone, testosterone, and androstenedione were increased only in early gestation (stages 1 and 2). Unlike month of year, gestation stage strongly predicted hormone concentration, but specific values to predict parturition date were not identified. Male testosterone and progesterone were higher in March-June (mating season) than July-January, while estradiol and androstenedione did not exhibit a seasonal pattern. Aquarium-managed cownose rays have similar reproductive patterns to what is reported in wild populations. Ultrasonographic monitoring with serial hormone analysis and accurate mating records will provide the most useful information for managing a reproductive population of cownose rays in an aquarium setting.

EVALUATION OF HEALTH PARAMETERS IN COWNOSE RAYS ( RHINOPTERA BONASUS) HOUSED IN A SEASONAL TOUCH POOL HABITAT COMPARED WITH AN OFF-EXHIBIT HABITAT.

Cownose rays ( Rhinoptera bonasus) are commonly displayed in zoo and aquarium touch pool exhibits; however, there is a gap in our understanding of how these practices might impact the health of these animals. The aim of this study was to evaluate and compare selected health parameters in cownose rays housed in a seasonal outdoor exhibit touch pool system with abundant public contact and an indoor off-exhibit holding system with minimal human contact. All animals underwent physical examination, ultrasound, cloacal wash and cytology, and blood collection for complete blood counts, point-of-care blood analysis, plasma protein electrophoresis, and plasma cholesterol electrophoresis in May and October 2014. Physical examination, ultrasound, and cloacal wash cytology findings were all unremarkable for both groups of animals. Significant differences in health parameters among animals by location and time point were few and included decreased heart rate ( F = 12.158, P = 0.001), increased lactate ( F = 6.838, P = 0.012), and increased low-density lipoproteins ( F = 19.961, P = 0.000) in touch pool animals over time. Based on these results, cownose rays inhabiting a touch pool exhibit and an off-exhibit system remained in comparable planes of health based on routine diagnostic modalities with few differences in measured health parameters.

WHOLE BLOOD TRANSFUSIONS TO TREAT SEVERE ANEMIA IN SEVEN COWNOSE RAYS ( RHINOPTERA BONASUS) AND ONE SHORT-TAIL STINGRAY ( DASYATIS BREVICAUDATA).

Blood transfusions can provide life-saving treatment to severely anemic animals. Due to limited availability and the difficulty of storing whole blood and blood products, such as fresh frozen plasma and packed red blood cells, exotic animals often receive fresh whole blood transfusions. Little is known about elasmobranch blood types and transfusions. Conspecific cross-matches within several different elasmobranch species were negative, indicating that in an emergency situation a single whole blood transfusion may be possible without causing a transfusion reaction. Experimental transfusions between healthy conspecific Atlantic rays ( Dasyatis sabina) showed no adverse reactions and autotransfusions in marbled electric rays ( Torpedo marmorata) were successful. There are no published reports of blood transfusions performed on clinically abnormal elasmobranchs. The following case series documents blood transfusions performed on seven cownose rays ( Rhinoptera bonasus) and one short-tail stingray ( Dasyatis brevicaudata). All rays were treated with the same protocol, which included pretreatment with steroids and antibiotics followed by an intravenous transfusion of freshly collected, heparinized, whole blood. Three animals survived and currently exhibit no abnormal clinical signs. Two animals died 55 days and 100 days post transfusion. Three animals died 2-22 days post transfusion. Although complications from blood transfusions could not be ruled out, all five animals that died had other health problems that likely contributed to their demise. All eight animals would almost certainly have died without a blood transfusion as they were severely anemic and moribund at the time of presentation. The methods described in this paper may be useful in the treatment of severely anemic elasmobranchs and this is the first report of blood transfusions in clinically abnormal elasmobranchs.

COMPARISON OF PRE- AND POSTQUARANTINE BLOOD CHEMISTRY AND HEMATOLOGY VALUES FROM WILD-CAUGHT COWNOSE RAYS (RHINOPTERA BONASUS).

Though one of the most widely kept elasmobranchs in human care, the cownose ray (CNR; Rhinoptera bonasus ), remains a species with minimal published information on hematologic reference intervals. As part of a larger study investigating the health and nutrition of the CNR, this study established a preliminary data set of plasma chemistry and hematology values specific to animals recently caught from the wild and compared this data set (intake sample) to values obtained following a period of quarantine (27-40 days) in an aquarium (exit sample). Blood samples were collected from 47 wild female (n = 46) and male (n = 1) CNR caught in pound nets off the coast of North Carolina and South Carolina. Differences between intake and exit values were analyzed. Due to the preponderance of female animals, data were not analyzed for sex differences. Plasma biochemical profiles were performed and analyzed. A select number of complete blood cell counts were performed (n = 24 from 12 animals). Statistically significant differences (P < 0.05) specific to time of sampling were determined for packed cell volume, total solids, blood urea nitrogen, sodium, chloride, potassium, phosphorus, cholesterol, glucose, and aspartate aminotransferase. Values reported are a significant expansion on the existing limited data for CNRs and will serve as a reference for health assessment of individuals both in the wild and in exhibit populations.

Case report: Management and long-term ophthalmic sequelae of monogenean ocular infestation in cownose rays (Rhinoptera bonasus).

Monogenean ectoparasitic flatworm infestations, particularly in closely confined populations, can result in severe epizootic disease that is often devastating and occasionally fatal. This case series describes a population of cownose rays (Rhinoptera bonasus) (n = 52) housed in an aquarium touch-tank exhibit that presented with severe ocular disease due to infestation with monogeneans, presumably Benedeniella posterocolpa of the Capsilidae family. The most severely affected individuals (n = 9), including several cases with bilaterally ruptured corneas, underwent serial examinations prior to and following treatment with praziquantel, systemic antibiotics, and corticosteroids. The entire population underwent frequent therapeutic water changes with a scheduled decrease in salinity, increase in temperature, and a series of seven praziquantel tank treatments. At the last follow up examination (3.75 years), the most common ocular findings were corneal fibrosis (18/18 eyes; 100%), cataract formation (13/18 eyes; 72.2%), synechia (8/18 eyes; 44.4%), and dyscoria (5/18 eyes; 27.8%). Despite severe corneal disease, including corneal rupture, all examined eyes (18/18; 100%) showed remarkable corneal remodeling and a largely clear visual axis. There are very few reports describing corneal disease in aquarium housed elasmobranchs, and no reports describe ophthalmic implications of monogenean infestation in these animals. This further underscores the importance of this case series in demonstrating the capacity for healing of elasmobranch eyes and can provide further guidance regarding prognosis and treatment in cases of severe corneal disease.

A Bioinspired Cownose Ray Robot for Seabed Exploration.

This article presents the design and the experimental tests of a bioinspired robot mimicking the cownose ray. These fish swim by moving their large and flat pectoral fins, creating a wave that pushes backward the surrounding water so that the fish is propelled forward due to momentum conservation. The robot inspired by these animals has a rigid central body, housing motors, batteries, and electronics, and flexible pectoral fins made of silicone rubber. Each of them is actuated by a servomotor driving a link inside the leading edge, and the traveling wave is reproduced thanks to the flexibility of the fin itself. In addition to the pectoral fins, two small rigid caudal fins are present to improve the robot's maneuverability. The robot has been designed, built, and tested underwater, and the experiments have shown that the locomotion principle is valid and that the robot is able to swim forward, perform left and right turns, and do floating or diving maneuvers.

PERUANOCOTYLE PELAGICA N. SP. (MONOGENEA: MONOCOTYLIDAE), PARASITE OF THE PACIFIC COWNOSE RAY RHINOPTERA STEINDACHNERI EVERMANN AND JENKINS, 1891 (BATOIDEA: RHINOPTERIDAE) FROM THE SOUTHERN MEXICAN PACIFIC OCEAN.

Peruanocotyle pelagica n. sp. is described based on specimens collected from the wall of the pharyngeal cavity of the Pacific cownose ray Rhinoptera steindachneri offshore Oaxaca and Guerrero, Mexico. The new species is distinguished from Peruanocotyle chisholmae by its anchors, which include a slender guard that curves towards the tip of the blade and which lack an accessory piece, morphological differences of the seminal vesicle, the lack of a male copulatory organ accessory piece and a greater number of spines, and an unsclerotized vagina. Molecular data of Peruanocotyle pelagica were generated to place the phylogenetic position of the genus within Monocotylidae.

Site-specific and density-dependent extinction of prey by schooling rays: generation of a population sink in top-quality habitat for bay scallops.

Bay scallops (Argopecten irradians concentricus) are patchily distributed on two dominant spatial scales: (1) geographically restricted to highly saline marine lagoons, and (2) locally abundant within such lagoons only in relatively discrete beds of seagrass habitat. In the Cape Lookout lagoonal system of North Carolina, adult bay scallop abundance in the most densely occupied seagrass bed (Oscar Shoal) exhibits repeatable declines from up to 70 m-2 to near zero in a 2- to 4-week period during late summer. This crash is completed before fall spawning can be initiated, thereby creating a population sink in what is the singly most productive patch of habitat. Field experiments conducted in the summers of 1996 and 1998 demonstrated that the seasonal extinction of bay scallops on Oscar Shoal can be prevented by the erection of 1-m2 stockades, made of 50-cm-high vertical poles, spaced every 25 cm, which inhibit access by cownose rays. Because these stockades were porous to emigration and physical transport, and open to access by all other predators of adult scallops, predation by migrating cownose rays is the only viable explanation for the crash. Consequently, the natural predation process in this system achieves the reproductive extinction of prey in the habitat patch of highest productivity. Over 7 years of observation, the mortality rate in this patch increased with summer density, reaching the asymptote of 100% at 10 m-2. The site-specific habitat selection by schools of rays may be based on prey density, which could render this example representative of a widespread generator of population sinks in habitat patches of high quality. The virtual extinction of scallops within Oscar Shoal despite nearby patches with relatively high density may be related to the highly efficient feeding behavior of schools and the high vulnerability of bay scallops in a context of multiple alternative prey types.

Behavioral responses of batoid elasmobranchs to prey-simulating electric fields are correlated to peripheral sensory morphology and ecology.

Electrosensory pore number, distribution, and sensitivity to prey-simulating electric fields have been described for many shark species. Electrosensory systems in batoids have received much less attention. Pore number and distribution have yet to be correlated to differences in sensitivity. However, pore number, pore distribution and sensitivity have been linked to behavior, diet, and morphology and follow species-specific trends. We report here that cownose rays have a greater number of pores than the yellow stingray, most of which are concentrated on the anterior ventral surface for both species. However, yellow stingrays have a broader arrangement of pores on both their dorsal and ventral surfaces than the cownose rays. Yellow stingrays demonstrated a median behavioral sensitivity to weak electric fields of 22nVcm(-1) and are among the most highly sensitive batoids studied to date. Cownose rays are less sensitive than all other elasmobranch species with a median sensitivity of 107nVcm(-1). As reported in previous studies, a higher pore number did not result in greater sensitivity. Cownose rays are benthopelagic schooling rays and may benefit from reduced sensitivity to bioelectric fields when they are surrounded by the bioelectric fields of conspecifics. Yellow stingrays, on the other hand, are typically solitary and bury in the substrate. A greater number of pores on their dorsal surface might improve detection of predators above them. Also, increased sensitivity and a broader distribution of pores may be beneficial as small prey items move past a buried ray.