Morfología comparativa de las células sanguíneas del tiburón nodriza Ginglymostoma cirratum (Bonnaterre, 1788) con diversos peces.

Se llevó a cabo una comparación entre la morfología de las células de la sangre periférica de 9 tiburones nodriza, de una raya americana y 9 peces teleósteos; además, se determinó la fórmula leucocitaria y la cantidad total de eritrocitos y leucocitos/mm 3 de sangre para cada pez estudiado. English Abstract This work is a comparation of the blood cells morphology between 9 nurse sharks, southern stingray and 9 bone fishes, moreover, it was maked a differential leucocytes count, red and white cells counts by blood mm3 for any fish.

Defensive strategies of neonate nurse sharks, Ginglymostoma cirratum, in an oceanic archipelago of the Western Central Atlantic

Two defensive tactics of neonates of nurse sharks, Ginglymostoma cirratum, are reported based on underwater observations. Described as "hiding behaviour" and "substrate resemblance", the defensive strategies were categorized according to the predominant habitat in which the individuals were found and to the behaviour displayed by the sharks in the presence of the observer. In structurally more complex habitats with a wide availability of shelters, the preferential behaviour displayed by neonates is to hide inside holes or crevices. When in open areas deprived of refuges, neonates tend to resemble arborescent coverings as seaweed banks or colonies of octocorals, which allows the use of more exposed habitats without increasing the susceptibility of capture by predators. Both aspects are relevant for a better understanding of the behaviour of neonates of G. cirratum and have important implications for identifying important habitat in nursery areas, and also for the management of this vulnerable species off South America.

Sequence analysis of complementary DNAs encoding C3 in the nurse shark ({\it Ginglymostoma cirratum\/})

Mammalian C3 is a pivotal complement protein, encoded for by a single gene. In some vertebrate species multiple C3 isoforms are products of different C3 genes. The goal of this study was to determine whether multiple genes encode for shark C3. A protocol was developed for the isolation of mRNA from shark blood for the isolation of C3 cDNA clones. RT-PCR amplification of mRNA, using sense (GCGEQNM) and antisense (TWLTAYV) primers encoding conserved regions of human C3, yielded 21 clones. The C3-like clones isolated shared 97% similarity with each other and 40% similarity to human C3. RACE-PCR amplification of shark liver RNA, using gene specific primers, yielded products ranging from 1800bp to 3000bp. Deduced amino acid sequence, corresponding to 408bp of the 1800bp fragment, was obtained which showed 51% similarity to human C3. These results suggest that nurse shark C3 might be encoded for by more than one gene. ^

Antibacterial proteins and peptides in nurse shark ( Ginglymostoma cirratum) peripheral blood leukocytes

In many vertebrate and invertebrate species mediators of innate immunity include antimicrobial peptides (AMPs) such as peptide fragments of histones and other proteins with previously ascribed different functions. Shark AMPs have not been described and this research examines the antibacterial activity of nurse shark (Ginglymostoma cirratum) peripheral blood leukocyte lysates. Screening of lysates prepared by homogenizing unstimulated peripheral blood leukocytes identified muramidase (lysozyme-like) and non-muramidase antibacterial activity. Lysates were tested for lysozyme using the lysoplate assays, and antibacterial (AB) activity was assayed for by a microdilution growth assay that was developed using Planococcus citreus as the target bacterium. Fractionation of crude lysates by ion exchange and affinity chromatography was followed by a combination of SDS-PAGE with LC/MS-MS and/or N-terminal sequence analysis of low molecular weight protein bands (<20 kDa). This yielded several peptides with amino acid sequence similarity to lysozyme, ubiquitin, hemoglobin, human histones H2A, H2B and H4 and to antibacterial histone fragments of the catfish and the Asian toad. Not all peptide sequences corresponded to peptides potentially antibacterial. The correlation of a specific protein band in active lysate fractions was accomplished by employing the acid-urea gel overlay assays in which AB activity was seen as zones of growth inhibition on a lawn of P. citreus at a position corresponding to that of the putative AB protein band. This study is the first to describe putative AMPs in the shark and their potential role in innate immunity.^

Construction and screening of A cDNA library for the C3 gene(s) of the Nurse Shark (Ginglymostoma Cirratum)

Mammalian C3 is a complement protein which consists of an α chain (125kDa) and β chain (75kDa) held together by a disulfide bond. The a chain contains a conserved thiolester site which provides the molecule with opsonic properties. The protein is synthesized as a single pro-C3 molecule which is post-translationally modified. C3 genes have been identified in organisms from different phyla, however, the shark C3 gene remains to be cloned. Sequence data from the shark will contribute to understanding further the evolution of this key protein. To obtain additional sequence data for shark C3 genes a cDNA library was constructed and screened with a DIG-labeled C3 probe. Fifty clones were isolated and sequenced. Analysis identified four sequences that yielded positive alignments with C3 of a variety of organisms including human C3. Deduced amino acid sequence analysis confirmed a β/α cut site (RRRR), the CR3 and properdin binding sites, the catalytic histidine, and the reactive thiolester sequence. In the shark there are at least two C3-like genes as the gene sequence obtained is distinct from that previously described.

Molecular characterization of GcC8 alpha, the functional homologue of human C8 alpha in the shark, Ginglymostoma cirratum

The focus of this study is to elucidate the components of the nurse shark (Ginglymostoma cirratum) membrane attack complex (MAC), specifically complement component C8a (GcC8u). Nurse shark C8a gene was cloned, sequenced, and analyzed and Western blot analysis performed to identify components of shark MAC. GcC8a consists of 2341 nucleotides that translate into a 589 amino acid sequence that shares 41.1% and 47.4 % identity with human and xenopus C8a, respectively. GcC8a conserves the MAC modular architecture and cysteine-rich backbone characteristic of complement proteins, including the cysteine residue that forms the C8a-y bond as well as the indel that is unique to C8a. Conservation of MAC protein structure is evident from crossreactivity of antihuman-MAC antibodies with shark serum proteins in Western blots which confirmed the presence of C8 and C9-like proteins in shark serum, however, did not resolve the question of whether C6 and/or C7 like proteins are present in shark.

Fish movement patterns in Virgin Islands National Park, Virgin Islands Coral Reef National Monument and adjacent waters

NOAA’s National Centers for Coastal Ocean Science (NCCOS)-Center for Coastal Monitoring and Assessment’s (CCMA) Biogeography Branch, National Park Service (NPS), US Geological Survey, and the University of Hawaii used acoustic telemetry to quantify spatial patterns and habitat affinities of reef fishes around the island of St. John, US Virgin Islands. The objective of the study was to define the movements of reef fishes among habitats within and between the Virgin Islands Coral Reef National Monument (VICRNM), the Virgin Islands National Park (VIIS), and Territorial waters surrounding St. John. In order to better understand species’ habitat utilization patterns among management regimes, we deployed an array of hydroacoustic receivers and acoustically tagged reef fishes. Thirty six receivers were deployed in shallow near-shore bays and across the shelf to depths of approximately 30 m. One hundred eighty four individual fishes were tagged representing 19 species from 10 different families with VEMCO V9-2L-R64K transmitters. The array provides fish movement information at fine (e.g., day-night and 100s meters within a bay) to broad spatial and temporal scales (multiple years and 1000s meters across the shelf). The long term multi-year tracking project provides direct evidence of connectivity across habitat types in the seascape and among management units. An important finding for management was that a number of individuals moved among management units (VICRNM, VINP, Territorial waters) and several snapper moved from near-shore protected areas to offshore shelf-edge spawning aggregations. However, most individuals spent the majority of their time with VIIS and VICRNM, with only a few wide-ranging species moving outside the management units. Five species of snappers (Lutjanidae) accounted for 31% of all individuals tagged, followed by three species of grunts (Haemulidae) accounting for an additional 23% of the total. No other family had more than a single species represented in the study. Bluestripe grunt (Haemulon sciurus) comprised 22% of all individuals tagged, followed by lane snappers (Lutjanus synagris) at 21%, bar jack (Carangoides ruber) at 11%, and saucereye porgy (Calamus calamus) at 10%. The largest individual tagged was a 70 cm TL nurse shark (Ginglymostoma cirratum), followed by a 65 cm mutton snapper (Lutjanus analis), a 47 cm bar jack, and a 41 cm dog snapper (Lutjanus jocu). The smallest individuals tagged were a 19 cm blue tang (Acanthurus coeruleus) and a 19.2 cm doctorfish (Acanthurus chirurgus). Of the 40 bluestriped grunt acoustically tagged, 73% were detected on the receiver array. The average days at large (DAL) was 249 (just over 8 months), with one individual detected for 930 days (over two and a half years). Lane snapper were the next most abundant species tagged (N = 38) with 89% detected on the array. The average days at large (DAL) was 221 with one individual detected for 351 days. Seventy-one percent of the bar jacks (N = 21) were detected on the array with the average DALs at 47 days. All of the mutton snapper (N = 12) were detected on the array with an average DAL of 273 and the longest at 784. The average maximum distance travelled (MDT) was ca. 2 km with large variations among species. Grunts, snappers, jacks, and porgies showed the greatest movements. Among all individuals across species, there was a positive and significant correlation between size of individuals and MDT and between DAL and MDT.

Composição de desembarque, conhecimento tradicional e risco de extinção: a captura de elasmobrânquios do estado da Paraíba, Brasil

Tese de mestrado em Ecologia Marinha, apresentada à Universidade de Lisboa, através da Faculdade de Ciências, 2016

Ecologia e conservação dos tubarões do arquipélago de Fernando de Noronha, com ênfase no tubarão-cabeça-de-cesto Carcharhinus perezi (Poey, 1876) (Carcharhiniformes, Carcharhinidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

A shark antibody heavy chain encoded by a nonsomatically rearranged VDJ is preferentially expressed in early development and is convergent with mammalian IgG

In most vertebrate embryos and neonates studied to date unique antigen receptors (antibodies and T cell receptors) are expressed that possess a limited immune repertoire. We have isolated a subclass of IgM, IgM1gj, from the nurse shark Ginglymostoma cirratum that is preferentially expressed in neonates. The variable (V) region gene encoding the heavy (H) chain underwent V-D-J rearrangement in germ cells (“germline-joined”). Such H chain V genes were discovered over 10 years ago in sharks but until now were not shown to be expressed at appreciable levels; we find expression of H1gj in primary and secondary lymphoid tissues early in life, but in adults only in primary lymphoid tissue, which is identified in this work as the epigonal organ. H1gj chain associates covalently with light (L) chains and is most similar in sequence to IgM H chains, but like mammalian IgG has three rather than the four IgM constant domains; deletion of the ancestral IgM C2 domain thus defines both IgG and IgM1gj. Because sharks are the members of the oldest vertebrate class known to possess antibodies, unique or specialized antibodies expressed early in ontogeny in sharks and other vertebrates were likely present at the inception of the adaptive immune system.

The functional roles of passive electroreception in non-electric fishes

Passive electroreception is a complex and specialised sense found in a large range of aquatic vertebrates primarily designed for the detection of weak bioelectric fields. Particular attention has traditionally focused on cartilaginous fishes, but a range of teleost and non-teleost fishes from a diversity of habitats have also been examined. As more species are investigated, it has become apparent that the role of electroreception in fishes is not restricted to locating prey, but is utilised in other complex behaviours. This paper presents the various functional roles of passive electroreception in non-electric fishes, by reviewing much of the recent research on the detection of prey in the context of differences in species' habitat (shallow water, deep-sea, freshwater and saltwater). A special case study on the distribution and neural groupings of ampullary organs in the omnihaline bull shark, Carcharhinus leucas, is also presented and reveals that prey-capture, rather than navigation, may be an important determinant of pore distribution. The discrimination between potential predators and conspecifics and the role of bioelectric stimuli in social behaviour is discussed, as is the ability to migrate over short or long distances in order to locate environmentally favourable conditions. The various theories proposed regarding the importance and mediation of geomagnetic orientation by either an electroreceptive and/or a magnetite-based sensory system receives particular attention. The importance of electroreception to many species is emphasised by highlighting what still remains to be investigated, especially with respect to the physical, biochemical and neural properties of the ampullary organs and the signals that give rise to the large range of observed behaviours.

New microsatellite loci for Carcharhinid sharks (Carcharhinus tilstoni and C. sorrah) and their cross-amplification in other shark species

Twelve microsatellite DNA markers were isolated in the spot-tail shark (Carcharhinus sorrah) and nine were isolated in Australian black-tip shark (Carcharhinus tilstoni). These loci plus 18 others developed for sharks from the genera Negaprion, Ginglymostoma, Carcharodon and Isurus were tested for amplification success on four species of Carcharhinus (including C. sorrah and C. tilstoni) and four other species representing three diverse families. Cross-amplification was most common within families. Five loci were subsequently tested for polymorphism on 50 C. sorrah and 60 C. tilstoni. The number of alleles per locus was two to 24 and the average heterozygosity was 0.54 (range 0.16-0.87) for C. sorrah and 0.64 (range 0.44-0.78) for C. tilstoni. These loci may be useful tools for genetic analyses of the Carcharhinidae.

A comunidade de tubarões (Chondrichthyes : selachii : Galea) da formação pirabas, Neógeno da Amazônia Oriental

A comunidade de tubarões (Selachii:Galea) proveniente da Formação Pirabas pode ser considerada como uma das mais representativas e de maior diversidade entre as unidades do Neógeno da América do Sul. A presença de 8 gêneros permitiu elaborar hipóteses sobre a reconstrução da cadeia trófica envolvendo outros elementos da paleofauna de vertebrados, assim como também serviu como um indicador paleoecológico adicional que corrobora dados anteriores sobre a reconstituição dos parâmetros paleoambientais da unidade geológica. Foi realizado uma revisão taxonômica das espécies previamente conhecidos para a Formação Pirabas, cuja composição da paleocomun idade de tubarões foi a seguinte: Carcharhinus spl Carcharhinus sp2, Carcharhinus sp3, Carcharhinus priscus, Sphyma magna, Hemipristis serra, Carcharodon megalodon, Isurus sp, Ginglyrnostoma serra, Ginglymostoma obliquum. Novas coletas possibilitaram a expansão de 31 exemplares depositados em museus brasileiros para 231 novos indivíduos, incluindo material microscópico. Esta coleta serviu para aumentar o conhecimento sobre a diversidade e aspectos paleoecológicos dos grupos representados.