Physiological and behavioral responses to hypoxia in the bonnethead shark, Sphyrna tiburo: routine swimming and respiratory regulation

We examined the effect of hypoxia on the swimming speed, respiration rate (oxygen uptake), gape and ventilation volume of the bonnethead shark, Sphyrna tiburo. We used a sonic flowmeter developed for this study to examine swimming speed changes of sharks held in artificial lagoons during diurnal dissolved oxygen changes. Sharks were observed to swim at about 34 cm s^-1 during the day but increased to about 40 cm s^-1 at night when dissolved oxygen levels fell to < 3 mg l^-1. Using a closed system respirometer we examined changes in swimming speed, respiration rate and gape at four dissolved oxygen levels. Swimming speeds averaged 24 to 25 cm s^-1 under normoxic conditions but increased to 38 to 40 cm s^-1 during hypoxia. Similarly, respiration rate increased with increasing speed and decreasing dissolved oxygen. Gape averaged about 1.0 cm under normoxic conditions and increased to a maximum of about 3.5 cm during hypoxia. Using assumed oxygen extraction efficiencies of 25, 50 and 75% and observed respiration rates, we estimated that ventilation volumes of about 25 to 470 l h^-1, depending upon oxygen concentration, would be necessary for gill ventilation. These experiments suggest that changes in swimming speed and mouth gape are important for respiratory regulation in ram ventilating sharks.     

Serum steroid concentrations and development of reproductive organs during puberty in male bonnethead sharks, Sphyrna tiburo

Puberty is a critical, hormone-mediated event during which an animal acquires the ability to breed and propagate. Despite the importance of this stage in animal reproduction, little is known regarding the physiological factors that regulate and/or accompany puberty in several vertebrate groups including elasmobranchs. To address the need for such information, the present study investigated morphological and hormonal changes that occur during puberty in male bonnethead sharks (Sphyrna tiburo). Serial changes in development of claspers, paired copulatory organs in male elasmobranchs, and serum steroid concentrations during puberty were evaluated in captive-held male S. tiburo. Captive-animal studies were supplemented by observations on gonadal development, gonaduct morphology, and serum steroid concentrations in feral, peripubertal male S. tiburo. Changes in size and histological architecture of testes and gonaducts of peripubertal sharks mirrored the seasonal progression of events that occur in these structures in mature males. Claspers grew in length continuously during puberty, but sharks did not reach functional maturity until a short period before mating activity commences in the mature population. Clasper growth appeared to be strictly regulated in S. tiburo, perhaps to ensure growth of these organs to sizes deemed critical for reproductive success. Serum concentrations of testosterone, dihydrotestosterone, progesterone, and 17β-estradiol increased in both captive and feral sharks during pubertal development, and may be associated with development of the gonads and gonaducts. Differences in hormone profiles of captive and feral sharks were observed at certain periods during puberty, but their origin remains unclear. This revised version was published online in July 2006 with corrections to the Cover Date.     

Hepatic cholangiocarcinoma and testicular mesothelioma in a wild-caught blue shark,Prionace glauca (L.)

An adult male blue shark, Prionace glauca (L.), caught in July 2000 by a recreational fisherman off Long Island, New York, USA, had a retained fishing hook from a previous capture. The hook penetrated the gastric wall and lacerated the right liver lobe. Macroscopic lesions consisted of transmural gastritis and peritonitis. Alteromonas sp. and Vibrio alginolyticus were isolated from the peritoneal fluid. In addition, a well delineated, sessile mass was found on the otherwise normal serosa of the right testis. Histopathological findings included mesothelial hyperplasia and hypertrophy involving diffusely the gastric, hepatic and parietal serosae, and forming a discrete testicular capsular mass compatible with mesothelioma. In the liver an intrahepatic cholangiocarcinoma, chronic hepatitis, biliary hyperplasia and increased numbers of melanomacrophages were found. In addition organisms compatible with histozoic and coelozoic myxosporeans were found within the skeletal muscle of the abdomen and intrahepatic bile ducts, respectively. This is the third literature report of a liver tumour and the first report of a coelomic mesothelioma from a shark.     

Standardized catch and survival rates,and effect of a ban on shark retention,Palau pelagic longline fishery

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Induction and duration of tonic immobility in the lemon shark,Negaprion brevirostris

Tonic immobility (TI) is an unlearned behavioral response characterized by a state of immobility and torpor. Effect of inter-trial interval on duration of tonic immobility was assessed in thirty, juvenile lemon sharks (Negaprion brevirostris). Regression analyses showed that massed trials of 12 per session increased the average duration of tonic immobility by 475 sec compared to spaced trials of 1 per session. Each experiment was composed of 24 trials. TI is stable and durations much longer in the lemon shark than for other sharks. These findings have enabled us to develop a quantitative bioassay for use in testing chemical shark repellents.     

Evaluation of the use of metallothionein as a biomarker for detecting physiological responses to mercury exposure in the bonnethead,Sphyrna tiburo

Previous studies have demonstrated that sharks, perhaps more so than any other fishes, are capable of bioaccumulating the non-essential toxic metal mercury (Hg) to levels that threaten the health of human seafood consumers. However, few studies have explored the potential effects of Hg accumulation in sharks themselves. Therefore, the goal of this study was to examine if physiological effects occur in sharks in response to environmentally relevant levels of Hg exposure. To address this goal, the relationship between muscle Hg concentrations and muscle/hepatic levels of metallothionein (MT), a widely used protein biomarker of toxic metal exposure in fish, was examined in bonnetheads, Sphyrna tiburo, from three Florida estuaries. Total Hg concentrations in bonnethead muscle, as determined using thermal decomposition and atomic absorption spectrometry, ranged from 0.22 to 1.78 μg/g wet weight and were correlated with animal size. These observations were consistent with earlier studies on Florida bonnetheads, illustrating that they experience bioaccumulation of Hg, often to levels that threaten the health of these animals or consumers of their meat. However, despite this, MT concentrations measured using Western blot analysis were not correlated with muscle Hg concentrations. These results suggest that either environmentally relevant levels of Hg exposure and uptake are below the physiological threshold for inducing effects in sharks or MT is a poor biomarker of Hg exposure in these fishes. Of these two explanations, the latter is favored based on a growing body of evidence that questions the use of MTs as specific indicators of Hg exposure and effects in fish.     

Epizootics of Streptococcus agalactiae infection in captive rays from Queensland,Australia

The aim of this study was to describe two epizootics of high mortalities from infection with Streptococcus agalactiae, occurring in captive rays held in a marine display aquarium in south‐east Queensland, Australia, in 2009 and 2010. Five different species of rays were affected, including mangrove whiprays (Himantura granulata), estuary rays (Dasyatis fluviorum), eastern shovelnose rays (Aptychotrema rostrata), white‐spotted eagle rays (Aetobatus narinari) and blue‐spotted mask rays (Neotrygon kuhlii). This report describes the history of both epizootics including collection, quarantine and husbandry of rays, the disease epizootics, clinico‐pathological features of the disease, antimicrobial therapy, autogenous vaccine production, and laboratory studies including clinical and histopathology, bacteriology, PCR, molecular serotyping and sequencing of the bacterium S. agalactiae.     

First record of Tetramicra brevifilum in lumpfish (Cyclopterus lumpus,L.)

A microsporidian species with 98.3–98.4% nucleotide identity to Tetramicra brevifilum (Journal of Fish Diseases, 3 , 1980, 495) was diagnosed in lumpfish (Cyclopterus lumpus, L.) broodstock held at a breeding and rearing facility in western Ireland. The fish were wild‐caught from the west coast of Ireland, and the first case was diagnosed one year after capture. Clinical signs included severe bloating, lethargy, exophthalmos, anorexia, white patches on the cornea and externally visible parasitic cysts on skin and fins. Necropsy revealed severe ascites, white nodules and vacuoles in all the internal organs and partial liquefaction of the skeletal muscle. On histological examination, microsporidian xenomas were observed in all internal organs, the skin, skeletal muscle, gills and the eyes. The microsporidian species was identified by molecular analysis and transmission electron microscopy. This is the first record of T. brevifilum infecting lumpfish, and the disease is considered to be of potential significance to the rising aquaculture industry of this species.     

Aeromonas jandaei and Aeromonas veronii caused disease and mortality in Nile tilapia,Oreochromis niloticus (L.)

Diseases caused by motile aeromonads in freshwater fish have been generally assumed to be linked with mainly Aeromonas hydrophila while other species were probably overlooked. Here, we identified two isolates of non‐A. hydrophila recovered from Nile tilapia exhibiting disease and mortality after exposed to transport‐induced stress and subsequently confirmed their virulence in artificial infection. The bacterial isolates were identified as Aeromonas jandaei and Aeromonas veronii based on phenotypic features and homology of 16S rDNA. Experimental infection revealed that the high dose of A. jandaei (3.7 × 10⁶ CFU fish^?1) and A. veronii (8.9 × 10⁶ CFU fish^?1) killed 100% of experimental fish within 24 h, while a 10‐fold reduction dose killed 70% and 50% of fish, respectively. When the challenge dose was reduced 100‐fold, mortality of the fish exposed to A. jandaei and A. veronii decreased to 20% and 10%, respectively. The survivors from the latter dose administration were rechallenged with respective bacterial species. Lower mortality of rechallenged fish (0%–12.5%) compared to the control groups receiving a primary infection (37.5%) suggested that the survivors after primary infection were able to resist secondary infection. Fish exposed to either A. jandaei or A. veronii exhibited similar clinical signs and histological manifestation.     

Induction of fertilizable eggs by conspecific vitellogenin implantation in captive female walking catfish,Clarias batrachus (Linn.)

The synthesis of vitellogenin (Vg) is induced by conspecific Vg (Vg1 and Vg2) and estradiol‐17β (E₂) as demonstrated by the pattern of ³H‐serine incorporation in the liver and plasma proteins. The incorporation studies indicated that the label was first incorporated into the liver after which it appeared in the blood in both E₂‐ and Vg‐treated male catfish. Since Vg was capable of inducing its own synthesis, experiments were conducted in females during preparatory–prespawning period (March–May) to make them gravid by implanting Vg pellets. Two implantations of 4 mg Vg1 pellets into female catfish with an interval of 15 days, followed by laboratory maintenance for 45 days of initial implantation showed a significant increment in ovarian weight with concomitant formation of yolky oocytes through synthesis and incorporation of Vg, whereas Vg2 implantation was not effective in this regard. Histological observation of yolky oocytes in Vg1‐treated group showed the peripheral migration of germinal vesicle (eccentric germinal vesicle), which indicates the onset of maturation. On 45th day, third implantation with 2 mg Vg pellets was performed and after 15 days, fish were hormonally induced with a single injection of hCG (2,000 IU/kg fish). Six groups were considered such as initial control, BSA‐implanted control, Vg1‐implanted, Vg2‐implanted, catfish collected from the field on the last day of the experiment and catfish collected during spawning period in this experiment with 3–7 fish in each group. Each of the experimental fish was sexually mature and the body weight was between 100 and 125 g. The percentage of ovulation and fertilization in the eggs of Vg1‐implanted group was 91% and 78%, respectively, which was almost similar to that of gravid female catfish collected during breeding period (July). The breeding performance in BSA‐ and Vg2‐treated females was very poor. The fertilized eggs were hatched in the laboratory conditions. Thus, in the female catfish, Vg1 not only induces vitellogenesis but also makes the oocytes viable for fertilization.     

Co‐Infection with Pseudomonas anguilliseptica and Delftia acidovorans in the European eel,Anguilla anguilla (L.): a case history of an illegally trafficked protected species

Inspections by customs agents at Barcelona airport discovered 420 kg of contraband glass eels prepared for shipment to Hong Kong. After confiscation of these animals by police, they were transported to holding facilities to be maintained until after a judicial hearing. Upon arrival, they were separated into two groups and held under ambient flow‐through conditions in fresh water. During their captivity period, several peaks in mortality occurred and multiple bacterial strains were isolated from moribund animals. Sequencing of 16S rDNA was used to determine specific identity of the isolates. An initial isolation of Pseudomonas anguilliseptica was treated with oxytetracycline. A subsequent isolation of Delftia acidovorans proved resistant to oxytetracycline and was treated with gentamicin in combination with sulphadiazine–trimethoprim. Once the health condition of the animals was stabilized, they were partitioned into groups and subsequently released as part of a restocking effort for the species following the guidelines of Regulation (EC) 1100/2007 (Anon 2007). This represents the first record for both bacterial species in the host Anguilla anguilla in the Spanish Mediterranean.