Nurse Shark Research Articles

Feature

Australia's Biodiversity Crisis – For all of Australia's weird and different organisms, the richness of the country's biodiversity has never been under greater threat. A Life Devoted to Botany – APBN speaks to the executive director of Sydney's Botanic Gardens Trust, Dr Tim Entwisle, about his passion for plant life and the challenges facing botany. Saving the Shark – There are less than 500 Grey Nurse Sharks off Australia's coasts and this species is in danger of slipping into extinction. Who's Who – William Burns tells APBN about the latest ongoings at top Australian research center, the Diamantina Institute for Cancer, Immunology and Metabolic Medicine. Trading Currency – Treat flora and fauna as economic currency to prevent their deterioration, says Australia 21. The Wark™: At the Interface of Nano and Bio-materials. Molecular Plant Breeding Cooperative Research Centre. Pressing On – Scientists at Sydney's Centenary Institute are working to improve health outcomes for the community at large. Redrawing the Map – Donald Hobern takes a look at the challenges of managing biodiversity data. Fostering the Good Drop – Businesses like Foster's Wine Estates show that they can thrive while committing themselves to stronger conservation standards.

Functional morphology of the feeding apparatus, feeding constraints, and suction performance in the nurse shark Ginglymostoma cirratum

The nurse shark, Ginglymostoma cirratum, is an obligate suction feeder that preys on benthic invertebrates and fish. Its cranial morphology exhibits a suite of structural and functional modifications that facilitate this mode of prey capture. During suction-feeding, subambient pressure is generated by the ventral expansion of the hyoid apparatus and the floor of its buccopharyngeal cavity. As in suction-feeding bony fishes, the nurse shark exhibits expansive, compressive, and recovery kinematic phases that produce posterior-directed water flow through the buccopharyngeal cavity. However, there is generally neither a preparatory phase nor cranial elevation. Suction is generated by the rapid depression of the buccopharyngeal floor by the coracoarcualis, coracohyoideus, and coracobranchiales muscles. Because the hyoid arch of G. cirratum is loosely connected to the mandible, contraction of the rectus cervicis muscle group can greatly depress the floor of the buccopharyngeal cavity below the depressed mandible, resulting in large volumetric expansion. Suction pressures in the nurse shark vary greatly, but include the greatest subambient pressures reported for an aquatic-feeding vertebrate. Maximum suction pressure does not appear to be related to shark size, but is correlated with the rate of buccopharyngeal expansion. As in suction-feeding bony fishes, suction in the nurse shark is only effective within approximately 3 cm in front of the mouth. The foraging behavior of this shark is most likely constrained to ambushing or stalking due to the exponential decay of effective suction in front of the mouth. Prey capture may be facilitated by foraging within reef confines and close to the substrate, which can enhance the effective suction distance, or by foraging at night when it can more closely approach prey.

First characterization of shark nursery habitat in the United States Virgin Islands: evidence of habitat partitioning by two shark species

Little is known of the diversity, demography, and essential fish habitat of sharks within the United States Virgin Islands (USVI) marine ecosystem. To examine species diversity and the relative abundance of elasmobranchs in this region, bottom-longline and hand-gear sampling was conducted in Fish Bay, St. John, USVI, from June 2004 to December 2005. In the 8 sampling trips during this period, 54 standardized longline sets caught 174 elasmobranchs comprising 5 species of sharks and 1 batoid. Overall catch per unit effort [ln(CPUE + 1) ± SE] was 1.83 ± 0.16 elasmobranchs 100 hooks–1 h–1. Lemon sharks Negaprion brevirostris had the highest relative abundance based on log-transformed CPUE data (0.98 ± 0.15), followed by blacktip sharks Carcharhinus limbatus (0.91 ± 0.18), southern stingrays Dasyatis americana (0.28 ± 0.08), nurse sharks Ginglymostoma cirratum (0.08 ± 0.05), blacknose sharks Carcharhinus acronotus (0.06 ± 0.04) and the Caribbean sharpnose Rhizoprionodon porosus (0.03 ± 0.03). The relative abundance of all species was significantly higher in the summer (2.6 ± 0.2) than during the winter (1.1 ± 0.2). For the blacktip (N = 89 captures of 74 individuals) and lemon (N = 66, 48 individuals) sharks, which comprised the bulk of the catch, mean fork length (± SE) was 51.9 ± 0.63 cm and 59.9 ± 1.2 cm, respectively, representing primarily neonatal and young-of-the-year life stages. The recapture rates for blacktip and lemon sharks were 21% and 29%, respectively, and nearly all recaptures occurred within the bay, indicating a high degree of site fidelity. Capture information and limited acoustic tracking provided evidence of spatial and temporal habitat partitioning by these 2 shark species within the bay. Although the CPUE of both species was highest over shallow (<1 m) seagrass substrate, lemon sharks were found and tracked exclusively on shallow, mangrove-fringed seagrass habitat, while blacktip sharks utilized a wider area of the bay. Fish Bay was determined to provide important nursery habitat for young juvenile lemon and blacktip sharks in the USVI.

Shark Attacks in Dakar and the Cap Vert Peninsula, Senegal: Low Incidence despite High Occurrence of Potentially Dangerous Species

BackgroundThe International Shark Attack File mentions only four unprovoked shark attacks on the coast of West Africa during the period 1828–2004, an area where high concentrations of sharks and 17 species potentially dangerous to man have been observed. To investigate if the frequency of shark attacks could be really low and not just under-reported and whether there are potentially sharks that might attack in the area, a study was carried out in Dakar and the Cap Vert peninsula, Senegal.Methodology/Principal FindingsPersonnel of health facilities, administrative services, traditional authorities and groups of fishermen from the region of Dakar were interviewed about the occurrence of shark attacks, and visual censuses were conducted along the coastline to investigate shark communities associated with the coasts of Dakar and the Cap Vert peninsula. Six attacks were documented for the period 1947–2005, including two fatal ones attributed to the tiger shark Galeocerdo cuvieri. All attacks concerned fishermen and only one occurred after 1970. Sharks were observed year round along the coastline in waters 3–15 m depth. Two species potentially dangerous for man, the nurse shark Ginglymostoma cirratum and the blacktip shark Carcharhinus limbatus, represented together 94% of 1,071 sharks enumerated during 1,459 hours of observations. Threatening behaviour from sharks was noted in 12 encounters (1.1%), including 8 encounters with C. limbatus, one with Galeocerdo cuvieri and 3 with unidentified sharks.Conclusions/SignificanceThese findings suggest that the frequency of shark attacks on the coast of West Africa is underestimated. However, they also indicate that the risk is very low despite the abundance of sharks. In Dakar area, most encounters along the coastline with potentially dangerous species do not result in an attack. Compared to other causes of water related deaths, the incidence of shark attack appears negligible, at least one thousand fold lower.

Characterization of constitutive antimicrobial activity associated with leukocyte lysates from the nurse shark, Ginglymostoma cirratum (44.42)

Abstract Antimicrobial peptides are mediators of innate immunity. Studies have shown that peptide fragments of histones, proteins primarily involved with DNA packaging, display antimicrobial activity. This project examines the antibacterial activity associated with shark leukocyte lysates and the potential role of histone fragments in constitutive innate immunity. Preliminary studies have demonstrated antimicrobial activity, distinct from that of lysozyme, in crude cell lysates prepared from unstimulated shark blood leukocytes. The antimicrobial activity has been partially purified using a combination of ion exchange, size exclusion and affinity chromatography. SDS PAGE analysis of fractions showed the presence of several low molecular weight (&amp;lt;15 kDa) protein bands. N-terminal amino acid sequence analysis of four peptides identified them as histone fragments with sequence similarity to mammalian histones H2A, H2B and H4. The peptide sequences are also similar to histone fragments described for the catfish and the Asian toad, which exhibit antibacterial activity. The spectrum of antibacterial activity, biochemical characterization and mode of action of shark histone fragments is under investigation. Such information will determine their potential use against clinically significant organisms. The research was supported in part by FIU’s Comparative Immunology Initiative (NIH R25GM061347).

Molecular cloning, structural analysis and expression of complement component Bf/C2 genes in the nurse shark, Ginglymostoma cirratum

Factor B and C2 are serine proteases that provide the catalytic subunits of C3 and C5 convertases of the alternative (AP) and classical (CP) complement pathways. Two Bf/C2 cDNAs, GcBf/C2-1 and -2 (previously referred to as nsBf/C2-A and nsBf/C2-B), were isolated from the nurse shark, Ginglymostoma cirratum. GcBf/C2-1 and -2 are 3364 and 3082 bp in length and encode a leader peptide, three CCPs, one VWFA, the serine protease domain and have a putative factor D/C1s/MASP cleavage site. Southern blots show that there might be up to two Bf/C2-like genes for each of the two GcBf/C2 isoforms. GcBf/C2-1 and -2 are constitutively expressed, albeit at different levels, in all nine tissues examined. Expression in erythrocytes is a novel finding. Structural analysis has revealed that the localization of glycosylation sites in the SP domain of both putative proteins indicates that the molecular organization of the shark molecules is more like C2 than factor B. Phylogenetic analysis indicates that GcBf/C2-1 and -2 and TrscBf of Triakis scyllia (another shark species) originated from a common ancestor and share a remote ancestor with Bf and C2 of mammals and bony fish.

Selection of cholera toxin specific IgNAR single-domain antibodies from a naïve shark library

Shark immunoglobulin new antigen receptor (IgNAR, also referred to as NAR) variable domains (Vs) are single-domain antibody (sdAb) fragments containing only two hypervariable loop structures forming 3D topologies for a wide range of antigen recognition and binding. Their small size (∼12 kDa) and high solubility, thermostability and binding specificity make IgNARs an exceptional alternative source of engineered antibodies for sensor applications. Here, two new shark NAR V display libraries containing >10 7 unique clones from non-immunized (naïve) adult spiny dogfish ( Squalus acanthias) and smooth dogfish ( Mustelus canis) sharks were constructed. The most conserved consensus sequences derived from random clone sequence were compared with published nurse shark ( Ginglymostoma cirratum) sequences. Cholera toxin (CT) was chosen for panning one of the naïve display libraries due to its severe pathogenicity and commercial availability. Three very similar CT binders were selected and purified soluble monomeric anti-CT sdAbs were characterized using Luminex 100 and traditional ELISA assays. These novel anti-CT sdAbs selected from our newly constructed shark NAR V sdAb library specifically bound to soluble antigen, without cross reacting with other irrelevant antigens. They also showed superior heat stability, exhibiting slow loss of activity over the course of one hour at high temperature (95 °C), while conventional antibodies lost all activity in the first 5–10 min. The successful isolation of target specific sdAbs from one of our non-biased NAR libraries, demonstrate their ability to provide binders against an unacquainted antigen of interest.

Shark-specific repeat sequences in multiple factor I-like genes in Ginglymostoma cirratum, the nurse shark

Shark-specific repeat sequences in multiple factor I-like genes in Ginglymostoma cirratum, the nurse shark

Construction of a nurse shark (Ginglymostoma cirratum) bacterial artificial chromosome (BAC) library and a preliminary genome survey.

BackgroundSharks are members of the taxonomic class Chondrichthyes, the oldest living jawed vertebrates. Genomic studies of this group, in comparison to representative species in other vertebrate taxa, will allow us to theorize about the fundamental genetic, developmental, and functional characteristics in the common ancestor of all jawed vertebrates.AimsIn order to obtain mapping and sequencing data for comparative genomics, we constructed a bacterial artificial chromosome (BAC) library for the nurse shark, Ginglymostoma cirratum.ResultsThe BAC library consists of 313,344 clones with an average insert size of 144 kb, covering ~4.5 × 1010 bp and thus providing an 11-fold coverage of the haploid genome. BAC end sequence analyses revealed, in addition to LINEs and SINEs commonly found in other animal and plant genomes, two new groups of nurse shark-specific repetitive elements, NSRE1 and NSRE2 that seem to be major components of the nurse shark genome. Screening the library with single-copy or multi-copy gene probes showed 6–28 primary positive clones per probe of which 50–90% were true positives, demonstrating that the BAC library is representative of the different regions of the nurse shark genome. Furthermore, some BAC clones contained multiple genes, making physical mapping feasible.ConclusionWe have constructed a deep-coverage, high-quality, large insert, and publicly available BAC library for a cartilaginous fish. It will be very useful to the scientific community interested in shark genomic structure, comparative genomics, and functional studies. We found two new groups of repetitive elements specific to the nurse shark genome, which may contribute to the architecture and evolution of the nurse shark genome.

Evoked Potential Audiograms of the Nurse Shark (Ginglymostoma cirratum) and the Yellow Stingray (Urobatis jamaicensis)

The hearing thresholds of the nurse shark, Ginglymostoma cirratum, and the yellow stingray, Urobatis jamaicensis, were measured using auditory evoked potentials (AEP). Stimuli were calibrated using a pressure-velocity probe so that the acoustic field could be completely characterized. The results show similar hearing thresholds for both species and similar hearing thresholds to previously measured audiograms for the lemon shark, Negaprion brevirostris, and the horn shark, Heterodontis francisi. All of these audiograms suggest poor hearing abilities, raising questions about field studies showing attraction of sharks to acoustic signals. By extrapolating the particle acceleration thresholds into estimates of their equivalent far-field sound pressure levels, it appears that these sharks cannot likely detect most of the sounds that have attracted sharks in the field.

Rapid isolation of IgNAR variable single-domain antibody fragments from a shark synthetic library

The immunoglobulin isotype IgNAR (Novel Antigen Receptor) was discovered in the serum of the nurse shark ( Ginglymostoma cirratum) and wobbegong shark ( Orectolobus maculates) as a homodimer of two protein chains, each composed of a single variable domain (V) domain and five constant domains. The IgNAR variable domain contains an intact antigen-binding site and functions as an independent domain able to react to antigen with both high specificity and affinity. Here we describe the successful construction of a synthetic phage-displayed library based upon a single anti-lysozyme clone HEL-5A7 scaffold, which was previously selected from an immune IgNAR variable domain library. The complementarity-determining region 3 (CDR3) loop of this clone was varied in both length and composition and the derived library was used to pan against two model proteins, lysozyme and leptin. A single anti-lysozyme clone (Ly-X20) and anti-leptin clone (Lep-12E1) were selected for further study. Both clones were shown to be functionally expressed in Escherichia coli, extremely thermostable and bind to corresponding antigens specifically. The results here demonstrate that a synthetic IgNAR variable domain library based on a single framework scaffold can be used as a route to generate antigen binders quickly, easily and without the need of immunization.

First molecular and biochemical analysis ofin vivoaffinity maturation in an ectothermic vertebrate

The cartilaginous fish are the oldest phylogenetic group in which Igs have been found. Sharks produce a unique Ig isotype, IgNAR, a heavy-chain homodimer that does not associate with light chains. Instead, the variable (V) regions of IgNAR bind antigen as soluble single domains. Our group has shown that IgNAR plays an integral part in the humoral response of nurse sharks (Ginglymostoma cirratum) upon antigen challenge. Here, we generated phage-displayed libraries of IgNAR V regions from an immunized animal and found a family of clones derived from the same rearrangement event but differentially mutated during expansion. Because of the cluster organization of shark Ig genes and the paucicopy nature of IgNAR, we were able to construct the putative ancestor of this family. By studying mutations in the context of clone affinities, we found evidence that affinity maturation occurs for this isotype. Subsequently, we were able to identify mutations important in the affinity improvement of this family. Because the family clones were all obtained after immunization, they provide insight into the in vivo maturation mechanisms, in general, and for single-domain antibody fragments.

Somatic Hypermutation and Junctional Diversification at Ig Heavy Chain Loci in the Nurse Shark

We estimate there are approximately 15 IgM H chain loci in the nurse shark genome and have characterized one locus. It consists of one V, two D, and one J germline gene segments, and the constant (C) region can be distinguished from all of the others by a unique combination of restriction endonuclease sites in Cmu2. On the basis of these Cmu2 markers, 22 cDNA clones were selected from an epigonal organ cDNA library from the same individual; their C region sequences proved to be the same up to the polyadenylation site. With the identification of the corresponding germline gene segments, CDR3 from shark H chain rearrangements could be analyzed precisely, for the first time. Considerable diversity was generated by trimming and N addition at the three junctions and by varied recombination patterns of the two D gene segments. The cDNA sequences originated from independent rearrangements events, and most carried both single and contiguous substitutions. The 53 point mutations occurred with a bias for transition changes (53%), whereas the 78 tandem substitutions, mostly 2-4 bp long, do not (36%). The nature of the substitution patterns is the same as for mutants from six loci of two nurse shark L chain isotypes, showing that somatic hypermutation events are very similar at both H and L chain genes in this early vertebrate. The cis-regulatory elements targeting somatic hypermutation must have already existed in the ancestral Ig gene, before H and L chain divergence.

Grey Nurse Shark Human Interactions and Portrayals: A Study of Newspaper Portrayals of the Grey Nurse Shark from 1969-2003

The content analysis of 41 Australian newspaper articles pertaining to the grey nurse shark.

Modulation in Feeding Kinematics and Motor Pattern of the Nurse Shark Ginglymostoma cirratum

Studies of feeding in bony fishes have almost universally demonstrated the ability of individuals to modulate their method of capture in response to differing stimuli. Preliminary evidence indicates that morphologically specialized inertial suction feeding sharks are the most likely fishes to lack inherent modulatory ability. We examined the ability of the nurse shark, Ginglymostoma cirratum, to modulate its feeding behavior based on different food types and sizes. G. cirratum is an inertial suction feeding fish that is apparently stereotyped in its food capture behavior. Electromyography showed no statistical difference between feeding motor patterns based on food type (squid or fish) or size (gape width or twice gape width), although there were slight inter-individual differences in the onset of muscle firing for some muscles. Kinematic analysis showed a statistical difference in variables associated with durations for different food types, with the durations for all variables being faster for squid bites than fish bites, but no difference based on the size of the food item. This apparent lack of modulation may be associated with specialization of the morphology and behavior of G. cirratum for obligate suction prey capture. This functional specialization constrains the method in which G. cirratum captures prey but does not appear to result in dietary specialization. An unusual post capture spit-suck manipulation allows this shark to handle and ingest large prey.

Variations in gastric acid secretion during periods of fasting between two species of shark

Vertebrates differ in their regulation of gastric acid secretion during periods of fasting, yet it is unknown why these differences occur. Elasmobranch fishes are the earliest known vertebrates to develop an acid secreting stomach and as such may make a good comparative model for determining the causative factors behind these differences. We measured gastric pH and temperature continuously during periods of fasting in captive free-swimming nurse sharks ( Ginglymostoma cirratum) using autonomous pH/temperature data-loggers. All nurse sharks secreted strong gastric acids (minimum pH 0.4) after feeding; however, for most of the sharks, pH increased to 8.2–8.7, 2–3 days after feeding. Half of the sharks also exhibited periodic oscillations in pH when the stomach was empty that ranged from 1.1 to 8.7 (acid secretion ceased for 11.3 ± 4.3 h day − 1 ). This is in contrast to the gastric pH changes observed from leopard sharks ( Triakis semifasciata) in a previous study, where the stomach remains acidic during fasting. The leopard shark is a relatively active, more frequently feeding predator, and continuous acid secretion may increase digestive efficiency. In contrast, the nurse shark is less active and is thought to feed less frequently. Periodic cessation of acid secretion may be an energy conserving mechanism used by animals that feed infrequently and experience extended periods of fasting.

Nitric oxide production by nurse shark ( Ginglymostoma cirratum) and clearnose skate ( Raja eglanteria) peripheral blood leucocytes

Reactive nitrogen intermediates, such as nitric oxide (NO), are important immunomodulators in vertebrate immune systems, but have yet to be identified as mediators of host defence in any member of class Chondrichthyes, the cartilaginous fishes. In the present study, production of NO by nurse shark ( Ginglymostoma cirratum) peripheral blood leucocytes (PBL) stimulated with bacterial cell wall lipopolysaccharide (LPS) was investigated. PBL were cultured for 24 to 96 h following stimulation with LPS at concentrations ranging from 0 to 25 μg ml −1, in both serum-supplemented and serum-free culture conditions. Production of NO was measured indirectly using the Griess reaction, with maximal NO production occurring after 72 h using 10% FBS and 10 μg LPS ml −1. Application of these culture conditions to PBL from another cartilaginous fish (clearnose skate, Raja eglanteria) resulted in a similar NO response. Addition of a specific inhibitor of inducible nitric oxide synthase (iNOS), l- N 6-(1-iminoethyl)lysine ( l-NIL), resulted in a significant decrease in the production of NO by PBL from both species.

Auditory evoked potential measurements in elasmobranchs

Auditory evoked potentials (AEP) were first used to examine hearing in elasmobranchs by Corwin and Bullock in the late 1970s and early 1980s, marking the first time AEPs had been measured in fishes. Results of these experiments identified the regions of the ear and brain in which sound is processed, though no actual hearing thresholds were measured. Those initial experiments provided the ground work for future AEP experiments to measure fish hearing abilities in a manner that is much faster and more convenient than classical conditioning. Data will be presented on recent experiments in which AEPs were used to measure the hearing thresholds of two species of elasmobranchs: the nurse shark, Ginglymostoma cirratum, and the yellow stingray, Urobatis jamaicencis. Audiograms were analyzed and compared to previously published audiograms obtained using classical conditioning with results indicating that hearing thresholds were similar for the two methods. These data suggest that AEP testing is a viable option when measuring hearing in elasmobranchs and can increase the speed in which future hearing measurements can be obtained.

Abundance and distribution of sharks in Los Roques Archipelago National Park and other Venezuelan oceanic islands, 1997–1998

The relative abundance and distribution of sharks caught by the artisanal fishery was determined for the most important offshore islands of Venezuela (Los Roques, Las Aves and La Blanquilla). Data were collected from January 1997 to October 1998, corresponding to 123 fishing days targeting sharks using bottom longline. We also present the results of 10 days of fishing activities targeting tunas and marlins by the same fleet using pelagic longline, in order to determine the importance of the shark bycatch in these fisheries around the offshore islands. A total of 15 shark species representing five families were captured during the study period. The catch composition and CPUE values expressed in weight and number showed that the most abundant species were Carcharhinus perezi and C. falciformis for Los Roques Archipelago and La Blanquilla Island, while C. perezi and Ginglymostoma cirratum were more common in Las Aves Archipelago. In Los Roques Archipelago, C. perezi and C. falciformis were most common during the second half of the year; a similar trend was observed for the rest of the shark species caught by the fishery. A positive relationship was observed between CPUE and fishing depth in the case of C. perezi and C. falciformis off Los Roques Archipelago: CPUE by weight increases with depth, while CPUE by number decreases with depth. The main factors that could be determining shark abundance and their spatial-temporal distribution are the characteristics of the ecosystem, depth, and biological characteristics such as birth, mating season and migratory patterns of the species.

Marine Reserve Design and Evaluation Using Automated Acoustic Telemetry: A Case-study Involving Coral Reef-associated Sharks in the Mesoamerican Caribbean

A non-overlapping acoustic receiver array was used to track the movements of two common shark species, nurse Ginglymostoma cirratum (n=25) and Caribbean reef Carcharhinus perezi (n=5), in and around Glover's Reef Marine Reserve (GRMR), off the coast of Belize, between May and October, 2004. Although both species exhibited partial site fidelity in that they were most likely to be detected near the area of original capture, both species also moved widely throughout the 10 by 30 km atoll. One Caribbean reef shark was detected by a monitor at Lighthouse Reef, 30 km from Glover's Reef across deep (&gt;400m) open water. The mean minimum linear dispersal (MLD) was 10.5 km for Caribbean reef sharks and 7.7 km for nurse sharks, with many individuals traveling more than the 10 km width of the no-take "conservation zone" of the marine reserve. Although most sharks were tagged within the conservation zone, individuals were detected outside this part of GRMR on average 48 days out of the 150 days of observations. However, of 7 nurse sharks tagged near the center of the conservation zone, 4 were never detected outside of this part of the reserve. In general, this study suggests that effective conservation of these large roving predators requires an ecosystem-based management approach including a zoned management plan, similar to that used at GRMR, in which a fairly large no-take reserve, incorporating diverse habitats and the connections between them, is surrounded by a larger area in which fishing is regulated.