Interações tróficas entre espécies pelágicas do Atlântico Sudoeste: utilizando isótopos estáveis e inferência bayesiana

Pós-graduação em Ciências Biológicas (Zoologia) - IBRC

Hábitos alimentares de jovens de tubarão-martelo Sphyrna lewini (Griffith & Smith, 1834) (Chondrichthyes, Sphyrnidae) no litoral do estado de São Paulo

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

Diets of Baird’s Beaked Whales, Berardius bairdii, in the Southern Sea Of Okhotsk and Off the Pacific Coast Of Honshu, Japan

Stomach contents were analyzed from 127 Baird’s beaked whales, Berardizls bairdii, taken in coastal waters of Japan. During late July-August of 1985- 1987, 1989, and 1991, 107 samples were collected from off the Pacific coast of Honshu. An additional 20 samples were collected from whales taken in the southern Sea of Okhotsk during late August-September of 1988 and 1989. Prey identification using fish otoliths and cephalopod beaks revealed the whales fed primarily on deep-water gadiform fishes and cephalopods in both regions. Prey species diversity and the percentage of cephalopods and fish differed between the two regions. Off the Pacific coast of Honshu the whales fed primarily on benthopelagic fishes (81.8%) and only 18.0% on cephalopods. Eight species of fish representing two families, the codlings (Moridae) and the grenadiers (Macrouridde), collectively made up 81.3% of the total. Thirty species of cephalopods representing 14 families made up 12.7%. In the southern Sea of Okhotsk, cephalopods accounted for 87.1% of stomach contents. The families Gonatidae and Cranchiidae were the predominant cephalopod prey, accounting for 86.7% of the diet. Gadiform fish accounted for only 12.9% of the diet. Longfin codling, Laernonma longipes, was the dominant fish prey in both regions. Depth distribution of the two commonly consumed fish off the Pacific coast of Honshu indicate the whales in this region fed primarily at depths ranging from 800 to 1,200 m.

POPULATION BIOLOGY AND DIET OF THE SOUTHERN KINGCROAKER Menticirrhus americanus (LINNAEUS, 1758) (PERCIFORMES: SCIAENIDAE) IN CARAGUATATUBA BAY, SOUTHEASTERN BRAZIL

This study evaluated the spatio-temporal distribution, population biology and diet of Menticirrhus americanus in Caraguatatuba Bay. Samples were taken monthly between August 2003 and October 2004, by trawling in two previously selected areas. The northern area is more exposed to wave activity and is influenced by a river, functioning as a small estuary. In contrast, the southern area is relatively sheltered from wave energy and influenced to a lesser degree by smaller rivers. The fishes' length was measured, and the sex and gonadal stage macroscopically identified. The abundance of this species was compared between areas and among months. The diet was identified and quantified. M. americanus occurred in equal proportions in the two study areas, being most abundant in April 2004, followed by December 2003 and January 2004. The population was dominated by small immature individuals. The few individuals in maturation or mature that were captured showed no seasonal pattern of distribution. This species had a varied diet, feeding on worms (nemerteans, sipunculans and echiurans), mollusks (bivalves and cephalopods), polychaetes, crustaceans and fish. The presence of intact nematodes in the intestine suggests that these are parasites. The results demonstrated that M. americanus has a homogeneous spatial and temporal distribution in Caraguatatuba Bay, being uniformly distributed between the south and north areas as well as across the months. This species can be considered a carnivorous predator, showing a preference for consuming benthic sandy-beach species such as glycerids and other polychaetes, crustaceans, and bivalve siphons.

FEEDING HABITS OF THE ATLANTIC SPOTTED DOLPHIN, Stenella frontalis, IN SOUTHEASTERN BRAZIL

This study presents new information on the feeding habits of the Atlantic spotted dolphins, Stenella frontalis, in the Western South Atlantic. Nine stomach contents of S. frontalis incidentally caught in fishing operations conducted by the gillnet fleet based on main harbour of Cananeia (25 degrees 00' S; 47 degrees 55'W), southeastern Brazil, were analyzed. These specimens were captured between 2005 and 2007. A total of 1 422 cephalopod beaks, 147 otoliths and three crustaceans were recovered from the stomach contents. The dolphins assessed preyed on at least eight different fish species of the families Trichiuridae, Carangidae, Sparidae, Merluccidae, Engraulidae, Sciaenidae, Congridae and Scombridae, five cephalopod species of the families Loliginidae, Sepiolidae, Tremoctopodidae and Thysanoteuthidae, and one shrimp species of the Penaeidae family. Based on the analysis of the Index of Relative Importance (IRI), the Atlantic cutlassfish, Trichiurus lepturus, was the most important fish species represented. Of the cephalopods, the squid Doryteuthis plei was by far the most representative species. Several items were reported for the first time as prey of the S. frontalis: Xiphopenaeus kroyeri, Tremoctopus violaceus, Semirossia tenera, Merluccius hubbsi, Pagrus pagrus and Paralonchurus brasiliensis. S. frontalis presented teuthophagous and ichthyofagous feeding habits, with apparent predominance of the first, and preyed mainly on pelagic and demersal items.

Revisiting a medical case of "stinging" in the human oral cavity caused by ingestion of raw squid (Cephalopoda: Teuthida): new data on the functioning of squid's spermatophores

Male squid produce intricate spermatophores that, when transferred to the female, undergo the spermatophoric reaction, a complex process of evagination that leads to the attachment of the spermatangium, that is, the everted spermatophore containing the sperm mass. While this process is still not completely understood, the medical literature includes several reports of "oral stinging" (i.e., punctured wounds in the human oral cavity) following consumption of raw male squid, which contains undischarged spermatophores able to inflict such wounds. Here, we revisit a recent medical report of oral stinging by Shiraki et al. (Pathol Int 61:749-751, 2011), providing an in-depth reanalysis of their histological biopsies and revealing vital information on the functioning of squid spermatophores. The morphology of the spermatangia attached within the oral cavity is similar to the condition found in spermatangia naturally attached to female squids. The spermatangia were able to superficially puncture the superficial layers of the oral stratified squamous epithelium, and numerous, minute stellate particles from the squid spermatophore were found adhered to the oral epithelium. These findings corroborate previous hypotheses on the functioning of squid spermatophores, namely that spermatophore attachment generally involves tissue scarification, and that stellate particles play a vital role in the attachment process. Moreover, spermatophore attachment is confirmed to be autonomous (i.e., performed by the spermatophore itself) in another squid species (possibly a loliginid), and the results strongly indicate that the attachment mechanism is not dependent upon a specialized epithelium, nor a mate's specific chemical stimulus. From the pathological point of view, the best prophylactic measure at present is the removal of the internal organs of the raw squid prior to its consumption.

Feeding habits of the atlantic spotted dolphin, Stenella frontalis, in southeastern Brazil

This study presents new information on the feeding habits of the Atlantic spotted dolphins, Stenella frontalis, in the Western South Atlantic. Nine stomach contents of S. frontalis incidentally caught in fishing operations conducted by the gillnet fleet based on main harbour of Cananéia (25°00'S; 47°55'W), southeastern Brazil, were analyzed. These specimens were captured between 2005 and 2007. A total of 1 422 cephalopod beaks, 147 otoliths and three crustaceans were recovered from the stomach contents. The dolphins assessed preyed on at least eight different fish species of the families Trichiuridae, Carangidae, Sparidae, Merluccidae, Engraulidae, Sciaenidae, Congridae and Scombridae, five cephalopod species of the families Loliginidae, Sepiolidae, Tremoctopodidae and Thysanoteuthidae, and one shrimp species of the Penaeidae family. Based on the analysis of the Index of Relative Importance (IRI), the Atlantic cutlassfish, Trichiurus lepturus, was the most important fish species represented. Of the cephalopods, the squid Doryteuthis plei was by far the most representative species. Several items were reported for the first time as prey of the S. frontalis: Xiphopenaeus kroyeri, Tremoctopus violaceus, Semirossia tenera, Merluccius hubbsi, Pagrus pagrus and Paralonchurus brasiliensis. S. frontalis presented teuthophagous and ichthyofagous feeding habits, with apparent predominance of the first, and preyed mainly on pelagic and demersal items.

Population biology and diet of the southern kingcroaker Menticirrhus americanus (Linnaeus, 1758) (Perciformes: Sciaenidae) in Caraguatatuba Bay, southeastern Brazil

This study evaluated the spatio-temporal distribution, population biology and diet of Menticirrhus americanus in Caraguatatuba Bay. Samples were taken monthly between August 2003 and October 2004, by trawling in two previously selected areas. The northern area is more exposed to wave activity and is influenced by a river, functioning as a small estuary. In contrast, the southern area is relatively sheltered from wave energy and influenced to a lesser degree by smaller rivers. The fishes' length was measured, and the sex and gonadal stage macroscopically identified. The abundance of this species was compared between areas and among months. The diet was identified and quantified. M. americanus occurred in equal proportions in the two study areas, being most abundant in April 2004, followed by December 2003 and January 2004. The population was dominated by small immature individuals. The few individuals in maturation or mature that were captured showed no seasonal pattern of distribution. This species had a varied diet, feeding on worms (nemerteans, sipunculans and echiurans), mollusks (bivalves and cephalopods), polychaetes, crustaceans and fish. The presence of intact nematodes in the intestine suggests that these are parasites. The results demonstrated that M. americanus has a homogeneous spatial and temporal distribution in Caraguatatuba Bay, being uniformly distributed between the south and north areas as well as across the months. This species can be considered a carnivorous predator, showing a preference for consuming benthic sandy-beach species such as glycerids and other polychaetes, crustaceans, and bivalve siphons.

Daily ration and feeding chronology of dominant diel vertical migrant fishes in the Subtropical Eastern North Atlantic Ocean

[EN] Diel Vertical Migrants (DVMs) are mainly zooplankton and micronekton which migrate upward from 400-500 m depth every night to feed on the productive epipelagic zone, coming back at dawn to the mesopelagic zone, where they defecate, excrete, and respire the ingested carbon. DVMs should contribute to the biological pump in the ocean and, accordingly, to the global CO2 balance. Although those migrants are mainly small fishes, cephalopods and crustaceans, the lanternfishes (myctophidae) usually contribute up to 80% of total DVMs biomass. Thus, myctophids may represent a pathway accounting for a substantial export of organic carbon to the deep ocean. However, the magnitude of this transport is still poorly known. In order to assess this active flux of carbon, we performed a preliminary study of mesopelagic organisms around the Canary Islands. Here we present the results of diet, daily rations and feeding chronology of Lobianchia dofleini, Hygophum hygomii and Ceratoscopelus maderensis, 3 dominant species of myctophids performing diel vertical migrations in the Subtropical Eastern North Atlantic Ocean. Samples were obtained on board the RV La Bocaina during June 2009. Myctophids were sorted and fixed in 4% buffered formalin and the stomach contents of target species were examined and weighted. Feeding chronology was approached by studying stomach fullness and state of digestion of prey items in individuals from hauls performed at different times and depths. Our results provide further information about lanternfishes feeding ecology in relation to their vertical migration patterns as well as their contribution to the biological carbon pump.

Identification, Abundance and Rearing of Mysids of Gran Canaria: aplication to laboratory and oceanographic respiratory metabolism studies

Programa de doctorado en Oceanografía

3D-electron microscopy of different conformations of cephalopod hemocyanins

Die Hämocyanine der Cephalopoden Nautilus pompilius und Sepia officinalis sorgen für den Sauerstofftransport zwischen den Kiemen und den Geweben. Sie bestehen aus einem zylindrischen Dekamer mit interner Kragenstruktur. Während eine Untereinheit (also eine Polypeptidkette) bei NpH aus sieben paralogen funktionellen Domänen (FU-a bis FU-g) besteht, führte ein Genduplikationsereignis der FU-d zu acht FUs in SoH (a, b, c, d, d´, e, f, g). In allen Mollusken Hämocyaninen bilden sechs dieser FUs den äußeren Ring und die restlichen die interne Kragenstruktur. rnrnIn dieser Arbeit wurde ein dreidimensionales Modell des Hämocyanins von Sepia officinalis (SoH) erstellt. Die Rekonstruktion, mit einer Auflösung von 8,8Å (FSC=0,5), erlaubt das Einpassen von Homolologiemodellen und somit das Erstellen eines molekularen Modells mit pseudo atomarer Auflösung. Des Weiteren wurden zwei Rekonstruktionen des Hämocyanins von Nautilus pompilius (NpH) in verschiedenen Oxygenierungszuständen erstellt. Die auf 10 und 8,1Å aufgelösten Modelle zeigen zwei verschiedene Konformationen des Proteins. Daraus ließ sich eine Modellvorstellung über die allosterische Funktionsweise ableiten. Die hier erreichte Auflösung von 8Å ist die momentan höchste eines Molluskenhämocyanins. rnAuf Grundlage des molekularen Modells von SoH konnte die Topologie des Proteins aufgeklärt werden. Es wurde gezeigt, dass die zusätzliche FU-d´ in den Kragen integriert ist und somit die prinzipielle Wandarchitektur aller Mollusken Hämocyanine identisch ist. Wie die Analyse des erstellten molekularen Modells zeigt werden sind die beiden Isoformen (SoH1 und SoH2) in den Bereichen der Interfaces nahezu identisch; auch der Vergleich mit NpH zeigt grosse Übereinstimmungen. Des weiteren konnte eine Fülle von Informationen bezüglich der allosterischen Signalübertragung innerhalb des Moleküls gewonnen werden. rnDer Versuch, NpH in verschiedenen Oxygenierungszuständen zu zeigen, war erfolgreich. Die Datensätze, die unter zwei atmosphärischen Bedingungen präpariert wurden, führten reproduzierbar zu zwei unterschiedlichen Rekonstruktionen. Dies zeigt, daß der hier entwickelte experimentelle Ansatz funktioniert. Er kann nun routinemäßig auf andere Proteine angewandt werden. Wie der strukturelle Vergleich zeigte, verändert sich die Orientierung der FUs durch die Oxygenierung leicht. Dies wiederum beeinflusst die Anordnung innerhalb der Interfaces sowie die Abstände zwischen den beteiligten Aminosäuren. Aus dieser Analyse konnte eine Modellvorstellung zum allosterischen Signaltransfer innerhalb des Moleküls abgeleitet werden, die auf einer Umordnung von Salzbrücken basiert.

3D-Strukturanalyse von Molluskenhämocyaninen aus elektronenmikroskopischen Bildern

Diese Arbeit präsentiert die bislang höchst aufgelösten KryoEM-Strukturen für ein Cephalopoden hämocyanin Dekamer (Nautilus pompilus Hämocyanin, NpH) und ein Gastropoden Hämocyanin Didekamer (keyhole limpet hemocyanin isoform 1). Durch die Methoden des “molecular modelling” und “rigid-body-fiting” wurde auch eine detaillierte Beschreibung beider Strukturen auf atomarem Niveau erstmalig möglich. Hämocyanine sind kupferhaltige Sauerstoff-Transportproteine die frei gelöst in Blut zahlreicher Arthropoden und Mollusken vorkommen. Allgemein sind Molluskenhämocyanine als Dekamere (Hohlzylinder aus 5 Untereinheiten-dimere) oder Didecamere (Zusammenlagerung von zwei Dekameren) zu finden. Durch Anlagerung weiterer Dekamere bilden sich teilweise tubuläre Multidekamere. Hämocyanine der Cephalopoden bestehen ausschließlich aus solitären Decameren. In Octopus und Nautilus bestehen die 10 Untereinheiten aus 7 funktionellen Einheiten(FU-a bis FU-g), wobei jede FU ein Sauerstoffmolekül binden kann. FUs a-f bilden die Wand des ringförmigen Moleküls und 10 Kopien der FU-g bilden einen sogenannten „inneren Kragenkomplex“. Das im Rahmen dieser Arbeit erstelltes molekulares Modell von NpH klärt die Struktur des Dekamers vollständig auf. Wir waren zum ersten Mal in der Lage das Untereinheiten-dimer, den Verlauf der Polypeptidkette und 15 unterschiedliche Kontaktstellen zwischen FUs zu identifizieren. Viele der inter-FU-Kontakte weisen Aminosäurenkonstellationen auf, die die Basis für die Übertragung allosterischer Wechselwirkungen zwischen FUs darstellen könnten und Hinweise für den Aufbau der allosterische Einheit geben. Potentielle Bindungsstellen für N-glykosidische Zucker und bivalente Kationen wurden auch identifiziert. Im Gegensatz zu NpH, kommen Gastropoden Hämocyanine (inkl. KLH) hauptsächlich als Didekamere vor und der Kragenkomplex wird in diesem Fall aus 2 FUs gebildet (Fu-g und FU-h). Die zusätzliche C'-terminale FU-h zeichnet sich durch eine spezielle Verlängerung von ~ 100 Aminosäuren aus. KLH stammt aus der kalifornische Schnecke Megathura crenulata und kommt seit mehreren Jahrzehnten als Immunostimulator in der immunologischen Grundlagenforschung und klinischen Anwendung zum Einsatz. KLH weist zwei Isoformen auf, KLH1 und KLH2. Das vorliegende Modell von KLH1 erlaubt die komplexe Architektur dieses riesigen Proteins in allen Details zu verstehen, sowie einen Vergleich zum dem NpH Dekamer auf atomare Ebene. Es wurde gefunden, dass das Untereinheitensegment a-b-c-d-e-f-g, sowie die equivalenten Kontaktstellen zwichen FUs stark konserviert sind. Dies deutet darauf hin, dass in Bezug auf die Übertragung allosterische Signale zwischen benachbarten FUs, grundlegende Mechanismen in beiden Molekülen beibehalten wurden. Weiterhin, konnten die Verbindungen zwischen den zwei Dekameren ertsmalig identifiziert werden. Schließlich, wurde die Topologie der N-glycosidischen Zucker, welche für die immunologische Eigenschaften von KLH1 von großer Bedeutung sind, auch aufgeklärt. Somit leistet die vorliegende Arbeit einen wesentlichen Schritt zum Verständnis der Quartärstruktur und Funktion der Molluskenhämocyanine.rn

The nematode parasite fauna in three odontocete species: Phocoena phocoena, Lagenorhynchus albirostris, Lagenorhynchus acutus from Northern Scotland and its relationships with diet composition

Food items and nematode parasites were identified from the stomachs of 42 individuals of Phocoena phocoena, 6 of Lagenorhynchus acutus and 8 of L. albirostris stranded off the coastal waters of Northern Scotland between 2004 and 2014. Post-mortem examinations have revealed heavy parasitic worm burdens. Four nematode species complex as Anisakis spp., Contracaeucum spp., Pseudoterronova spp., and Hysterothylacium spp. were recorded. Data on presence of the anisakid species in cetaceans, reported a significative relationship between the presence of Hysterothylacium and the month of host stranding; suggesting a decrease of larval H. aduncum abundance in the period between April and August due to a seasonal effect related to prey availability. Similarly, the parasite burden of the all anisakid genera was related to the year fraction of stranding, and a relationship statistically significant was found just for L. albirostris with an increase between April and October. This finding is explained by a seasonality in occurrence of white-beaked dolphins, with a peak during August, that might be related to movements of shared prey species and competition with other species (Tursiops truncatus). Geographical differences were observed in parasites number of all anisakid species, which was the highest in cetaceans from the East area and lowest in the North coast. The parasites number also increased significantly with the length of the animal and during the year, but with a significant seasonal pattern only for P. phocoena. Regarding diet composition, through a data set consisting of 34 harbour porpoises and 1 Atlantic white-sided dolphins, we found a positive association between parasite number and the cephalopods genus Alloteuthis. This higher level of parasite infection in squid from this area, is probably due to a quantitative distribution of infective forms in squid prey, an abundance of the final host and age or size maturity of squid.

(Fig. 3) Sepia officinalis standard metobolic rate and seawater pH (NBS) during experiments, 2008

Ocean acidification and associated changes in seawater carbonate chemistry negatively influence calcification processes and depress metabolism in many calcifying marine invertebrates. We present data on the cephalopod mollusc Sepia officinalis, an invertebrate that is capable of not only maintaining calcification, but also growth rates and metabolism when exposed to elevated partial pressures of carbon dioxide (pCO2). During a 6 wk period, juvenile S. officinalis maintained calcification under ~4000 and ~6000 ppm CO2, and grew at the same rate with the same gross growth efficiency as did control animals. They gained approximately 4% body mass daily and increased the mass of their calcified cuttlebone by over 500%. We conclude that active cephalopods possess a certain level of pre-adaptation to long-term increments in carbon dioxide levels. Our general understanding of the mechanistic processes that limit calcification must improve before we can begin to predict what effects future ocean acidification will have on calcifying marine invertebrates.

Adverse effects of ocean acidification on early development of squid (Doryteuthis pealeii)

Anthropogenic carbon dioxide (CO2) is being absorbed into the ocean, altering seawater chemistry, with potentially negative impacts on a wide range of marine organisms. The early life stages of invertebrates with internal and external aragonite structures may be particularly vulnerable to this ocean acidification. Impacts to cephalopods, which form aragonite cuttlebones and statoliths, are of concern because of the central role they play in many ocean ecosystems and because of their importance to global fisheries. Atlantic longfin squid (Doryteuthis pealeii), an ecologically and economically valuable taxon, were reared from eggs to hatchlings (paralarvae) under ambient and elevated CO2 concentrations in replicated experimental trials. Animals raised under elevated pCO2 demonstrated significant developmental changes including increased time to hatching and shorter mantle lengths, although differences were small. Aragonite statoliths, critical for balance and detecting movement, had significantly reduced surface area and were abnormally shaped with increased porosity and altered crystal structure in elevated pCO2-reared paralarvae. These developmental and physiological effects could alter squid paralarvae behavior and survival in the wild, directly and indirectly impacting marine food webs and commercial fisheries.