xix, 166 p. : ill. / The nemertean worm Carcinonemertes errans is an egg predator on the Dungeness crab, Cancer magister, an important fishery species along the west coast of North America. This study examined the estuarine distribution and larval biology of C. errans. Parasite prevalence and mean intensity of C. errans infecting C. magister varied along an estuarine gradient in the Coos Bay, Oregon. Crabs nearest the ocean carried the heaviest parasite loads, and larger crabs were more heavily infected with worms. Seasonal infection patterns were seen at some sites within the bay. Crabs from coastal waters carried significantly more worms than did crabs from the bay, suggesting that the estuary may be acting as a parasite refuge for estuarine crabs. In laboratory experiments, C. errans all died in salinities below 10 within 6 days, but C. errans showed some tolerance to salinities 20 and above. These results suggest that salinity alone does not likely account for the estuarine gradient of C. errans in Coos Bay. Larvae of C. errans raised from hatching never settled in the laboratory. Competent larvae taken in plankton tows were morphologically distinct from larvae raised in laboratory cultures and did settle successfully on C. magister under laboratory conditions. Initial settlement was reversible within a 24-hour window. After 48 hours, a non-reversible metamorphosis occurred wherein worms lost one pair of eyes and the propensity to swim. In field settlement experiments, C. errans was capable of infecting hosts from the water column and preferred to settle on crabs already infected with juvenile worms, although this preference was density dependent. In monthly plankton tows, larvae of C. errans were found only between August and November, suggesting a long larval life for this species. Larvae did not feed under laboratory conditions, nor did they absorb dissolved organics. When exposed to a natural angular light distribution, larvae of C. errans were rarely photopositive. Larvae were most sensitive to blue-green light. Low intensity light invoked a photonegative response. Larvae were geopositive at hatching but geonegative thereafter. / Committee in charge: Brendan Bohannan, Chairperson; Craig Young, Advisor; Svetlana Maslakova, Member; Alan Shanks, Member; William Orr Outside, Member

Entre os meses de junho e agosto 2006 dois surtos de intoxicação pelas larvas de P. flavipes ocorreram, um em ovinos (surto 1) e outro em bovinos (surto 2). O surto nos ovinos ocorreu nos meses de junho e julho no município de Encruzilhada do Sul RS e morreram 25 ovinos de um rebanho de 175 ovinos e 11 bovinos. Os ovinos eram mantidos em uma área de 40 hectares de pasto nativo. O surto nos bovinos ocorreu no mês de agosto no município de Sombrio SC e morreram 17 animais que estavam num lote de 77 bovinos, o rebanho total da propriedade era de 280 animais. O lote de bovinos estava em uma área de 90 hectares onde dois animais morreram devido à ingestão das larvas de P. flavipes. O lote foi transferido para outra área onde morreram mais 15 animais em um período de 5 dias. Seis ovinos do surto 1 e seis bovinos do surto 2 foram necropsiados. Grande quantidade de agrupamentos de larvas de P. flavipes foram encontradas no campo e no rúmen dos animais de ambos os surtos. Larvas de P. flavipes foram coletadas em ambos os surtos e foram armazenadas congeladas a -20º. Larvas frescas e larvas congeladas foram administradas para 6 ovinos, por meio de uma seringa com a ponta cortada. A menor dose letal foi de 7,5 g/kg em administração única. Animais que receberam doses sub-letais de 5 g/kg e posteriormente doses letais de 10 g/kg e 15 g/kg em intervalos de 15 e 30 dias, mostraram menor sensibilidade à intoxicação, um animal não adoeceu e outro adoeceu apenas depois de receber uma dose de 15 g/kg. Os animais intoxicados experimentalmente mostraram sinais de doença cerca de 48 horas e morreram cerca de 54 horas após a intoxicação. O exame bioquímico realizado em intervalos de 12 horas revelou alterações apenas nos animais que adoeceram. Os níveis séricos de GGT apresentavam-se elevados depois de 24 horas da intoxicação e continuaram a se elevar até a morte, AST apresentou aumento significativo cerca de 30 horas após a dosificação e posteriormente decaindo até a morte, os níveis séricos de glicose sofreram queda próximo à morte dos animais. As lesões de necropsia observadas nos casos espontâneos e experimentais foram semelhantes e mais consistentes no fígado, que se apresentava com acentuação do padrão lobular e com áreas de coloração amarelada com petéquias subcapsulares. Foram observados também edemas cavitários, edema da parede da vesícula biliar, perirrenal e na região inicial do duodeno, pâncreas e abomaso. Além das hemorragias no fígado havia hemorragias no tecido subcutâneo, coração, e mucosas e serosas da cavidade abdominal. O principal achado histopatológico era caracterizado por necrose coagulativa centrolobular ou massiva associada à hemorragia e congestão centrolobular e degeneração e tumefação hepática na região periportal. Observou-se ainda, depleção e necrose linfóide nos centros germinativos de linfonodos, nas placas de Peyer e na polpa branca do baço. A microscopia eletrônica demonstrou lesão hepática com hepatócitos necróticos, e dilatação dos espaços de Disse e endotélio vascular. Foi observada também intensa proliferação de reticulo endoplasmático liso no animal que recebeu doses graduais das larvas. As larvas descongeladas e dessecadas a 100ºC por 24 horas foram administradas a um coelho e mostraram-se tóxicas. Outros dois coelhos receberam frações sólidas e liquidas das larvas respectivamente e apenas o coelho que recebeu a fração sólida morreu. / Between June and August 2006, two outbreaks of P. flavipes larvae poisoning were observed. In outbreak 1 occurred in a farm located at the county of Encruzilhada do Sul, Rio Grande do Sul State, Brazil, 25 out of 175 sheep were affected and died. Although there were 11 cattle in the same paddock, none of them was affected. Animals were kept in a 40 hectares paddock of native pastures. In outbreak 2, occurred in a farm located at county of Sombrio, Santa Catarina State, 11 out of 77 cattle were affected and died. In total, 6 sheep and 6 cattle from respective outbreaks were necropsied. High numbers of compact masses containing up to 150 larvae were scattered on the paddocks in which animals were grazing. Larvae were collected and frozen at –20ºC. Perreyia flavipes larval body fragments and heads were found in the forestomach contents of all necropsied animals. Fresh and thawed larvae were administered to six sheep by a tip-cut plastic syringe. The lower single lethal dosis was 7,5 g/kg. Animals which received an initial sublethal doses of 5 g/kg and subsequently were dosed with lethal 10 to 15 g/kg at 15 and 30 days intervals showed lower susceptibility to the intoxication. Animals that were experimentally poisoned showed signs of disease about 48 h after dosing and died in approximately 54 h. Biochemical tests performed at 12 h intervals showed changes only in diseased animals. Serum levels of GGT started being higher 24 h after intoxication and kept enhancement until death. AST serum levels were significatively enhanced about 30 h after dosing and then, decreased to death. Glucose serum levels decreased close to the death. Necropsy lesions were similar in both, spontaneous and experimental cases and were most prominent in the livers, which had enhanced lobular pattern and yellowish areas with subcapsular pinpoint hemorrhages. Edema in body cavities, gallbladder wall, perirenal tissues, initial portion of duodenum, pancreas, and abomasum was also seen. Hemorrhages were also present in subcutaneous tissues, heart, and in mucosal and serosal membranes of the abdominal cavity. The principal histological finding was centrolobular to massive coagulative necrosis associated with centrolobular hemorrhage and congestion and periportal degeneration and tumefaction. There also were lymphoid depletion and necrosis in the germinative centres of lymph nodes, Peyer’s patches, and white pulp of the spleen. Transmission electron microscopy showed necrotic hepatocytes and dilatation in the space of Disse and vascular endothelium. Severe proliferation of smooth endoplasmic reticulum was also seen in animal receiving more than one dosis of larvae. Thawed larvae were desiccated at 100ºC for 24 h and administered to one rabbit that also became ill. Additional two rabbit were dosed with solid or liquid fractions of larvae and only which received the solid fraction died.

O carrapato bovino Boophilus microplus está presente em áreas tropicais e subtropicais no mundo e é um ectoparasito hematófago que causa inúmeras perdas à bovinocultura, através da espoliação ao bovino ou das doenças que transmite como vetor. Atualmente, o principal método de controle empregado baseia-se em produtos químicos, que são onerosos e contaminam o ambiente. Estudos a respeito da ecologia, comportamento e fisiologia de B. microplus são importantes para o desenvolvimento de novos métodos de controle do carrapato. A resposta ao estresse oxidadivo e a morte celular programada, examinadas neste estudo, permitem compreender melhor os mecanismos fisiológicos usados pelo carrapato adulto e seus ovos e larvas para sobreviver no ambiente. A resposta a estresse oxidativo foi analisada em ovos e larvas. A atividade de GST e outras moléculas envolvidas em mecanismos de proteção contra estresse oxidativo variaram dependendo do tempo transcorrido após a postura e eclosão. A cinética do consumo de oxigênio apresentou correlação positiva com o aumento na atividade de GST durante a embriogênese. Um alto conteúdo de substâncias reativas ao ácido tiobarbitúrico foi observado em extratos de ovos e larvas, indicando que carrapatos apresentam alto estresse oxidativo durante a embriogênese e desenvolvimento larval. Além disso, a atividade de GST apresentou forte correlação com peroxidação de lipídeos, uma indicação de seu papel na defesa antioxidante em ovos. Com o objetivo de melhor caracterizar o processo de morte celular, que elimina tecidos não mais necessários e leva à degeneração de glândulas salivares, ovários e singânglios, foi feita uma investigação usando ensaios de visualização de fragmentação de DNA em gel de agarose, teste cometa e TUNEL e a ativação da via apoptótica foi analisada usando ensaio de caspase. A fragmentação de DNA e a atividade enzimática de caspase-3 foram observadas em glândulas salivares e ovários 48 e 72 h após a remoção do carrapato do hospedeiro; em singânglios estes parâmetros foram mantidos em baixos níveis após 48 h. Estes resultados obtidos sugerem que há um controle refinado de manutenção de tecido através de apoptose. / The cattle tick Boophilus microplus is present in tropical and subtropical areas in the world and it is a haematophagous ectoparasite that causes several losses to cattle breeding, through the bovine exploiting and diseases transmitted. Currently, the main control method is based on chemicals, which are expensive and contaminate the environment. Studies about ecology, behavior and physiology of B. microplus are important to develop new methods for the tick control. The data examined in this study allow understanding the physiologic mechanisms used by the tick, eggs and larvae to survive in the environment. For this, the oxidative stress response in eggs and larvae and the cell death in different tissues of adult tick females during the pre-oviposition period were analyzed. The oxidative stress response was analyzed in eggs and larvae and the results showed that the GST activity and other molecules involved in mechanism of protection against oxidative stress varied depending on the time elapsed after oviposition and eclosion. A positive correlation was observed between the oxygen consumption kinetics and the increase in GST activity during embryogenesis. A high content of thiobarbituric acid reactive substances were observed in egg and larva extracts, indicating that ticks face high oxidative stress during embryogenesis and aging. In addition, GST activity presented strong positive correlation with lipid peroxidation, an indication that it plays a role in oxidant defences in eggs. In order to better characterize the cell death process that eliminates unnecessary tissues, the degeneration of salivary glands, ovaries and synganglia was investigated using DNA fragmentation in agarose gel, comet and TUNEL assays, and apoptosis activation pathway by the caspase assay. DNA fragmentation and enzymatic activity of caspase-3 were observed in salivary glands and ovaries at 48 and 72 h after tick removal from the host; in synganglia these parameters were maintained at low levels upon 48 h. These results obtained suggest that there is a refined control of tissue maintenance through apoptosis.

Entre os meses de junho e agosto 2006 dois surtos de intoxicação pelas larvas de P. flavipes ocorreram, um em ovinos (surto 1) e outro em bovinos (surto 2). O surto nos ovinos ocorreu nos meses de junho e julho no município de Encruzilhada do Sul RS e morreram 25 ovinos de um rebanho de 175 ovinos e 11 bovinos. Os ovinos eram mantidos em uma área de 40 hectares de pasto nativo. O surto nos bovinos ocorreu no mês de agosto no município de Sombrio SC e morreram 17 animais que estavam num lote de 77 bovinos, o rebanho total da propriedade era de 280 animais. O lote de bovinos estava em uma área de 90 hectares onde dois animais morreram devido à ingestão das larvas de P. flavipes. O lote foi transferido para outra área onde morreram mais 15 animais em um período de 5 dias. Seis ovinos do surto 1 e seis bovinos do surto 2 foram necropsiados. Grande quantidade de agrupamentos de larvas de P. flavipes foram encontradas no campo e no rúmen dos animais de ambos os surtos. Larvas de P. flavipes foram coletadas em ambos os surtos e foram armazenadas congeladas a -20º. Larvas frescas e larvas congeladas foram administradas para 6 ovinos, por meio de uma seringa com a ponta cortada. A menor dose letal foi de 7,5 g/kg em administração única. Animais que receberam doses sub-letais de 5 g/kg e posteriormente doses letais de 10 g/kg e 15 g/kg em intervalos de 15 e 30 dias, mostraram menor sensibilidade à intoxicação, um animal não adoeceu e outro adoeceu apenas depois de receber uma dose de 15 g/kg. Os animais intoxicados experimentalmente mostraram sinais de doença cerca de 48 horas e morreram cerca de 54 horas após a intoxicação. O exame bioquímico realizado em intervalos de 12 horas revelou alterações apenas nos animais que adoeceram. Os níveis séricos de GGT apresentavam-se elevados depois de 24 horas da intoxicação e continuaram a se elevar até a morte, AST apresentou aumento significativo cerca de 30 horas após a dosificação e posteriormente decaindo até a morte, os níveis séricos de glicose sofreram queda próximo à morte dos animais. As lesões de necropsia observadas nos casos espontâneos e experimentais foram semelhantes e mais consistentes no fígado, que se apresentava com acentuação do padrão lobular e com áreas de coloração amarelada com petéquias subcapsulares. Foram observados também edemas cavitários, edema da parede da vesícula biliar, perirrenal e na região inicial do duodeno, pâncreas e abomaso. Além das hemorragias no fígado havia hemorragias no tecido subcutâneo, coração, e mucosas e serosas da cavidade abdominal. O principal achado histopatológico era caracterizado por necrose coagulativa centrolobular ou massiva associada à hemorragia e congestão centrolobular e degeneração e tumefação hepática na região periportal. Observou-se ainda, depleção e necrose linfóide nos centros germinativos de linfonodos, nas placas de Peyer e na polpa branca do baço. A microscopia eletrônica demonstrou lesão hepática com hepatócitos necróticos, e dilatação dos espaços de Disse e endotélio vascular. Foi observada também intensa proliferação de reticulo endoplasmático liso no animal que recebeu doses graduais das larvas. As larvas descongeladas e dessecadas a 100ºC por 24 horas foram administradas a um coelho e mostraram-se tóxicas. Outros dois coelhos receberam frações sólidas e liquidas das larvas respectivamente e apenas o coelho que recebeu a fração sólida morreu. / Between June and August 2006, two outbreaks of P. flavipes larvae poisoning were observed. In outbreak 1 occurred in a farm located at the county of Encruzilhada do Sul, Rio Grande do Sul State, Brazil, 25 out of 175 sheep were affected and died. Although there were 11 cattle in the same paddock, none of them was affected. Animals were kept in a 40 hectares paddock of native pastures. In outbreak 2, occurred in a farm located at county of Sombrio, Santa Catarina State, 11 out of 77 cattle were affected and died. In total, 6 sheep and 6 cattle from respective outbreaks were necropsied. High numbers of compact masses containing up to 150 larvae were scattered on the paddocks in which animals were grazing. Larvae were collected and frozen at –20ºC. Perreyia flavipes larval body fragments and heads were found in the forestomach contents of all necropsied animals. Fresh and thawed larvae were administered to six sheep by a tip-cut plastic syringe. The lower single lethal dosis was 7,5 g/kg. Animals which received an initial sublethal doses of 5 g/kg and subsequently were dosed with lethal 10 to 15 g/kg at 15 and 30 days intervals showed lower susceptibility to the intoxication. Animals that were experimentally poisoned showed signs of disease about 48 h after dosing and died in approximately 54 h. Biochemical tests performed at 12 h intervals showed changes only in diseased animals. Serum levels of GGT started being higher 24 h after intoxication and kept enhancement until death. AST serum levels were significatively enhanced about 30 h after dosing and then, decreased to death. Glucose serum levels decreased close to the death. Necropsy lesions were similar in both, spontaneous and experimental cases and were most prominent in the livers, which had enhanced lobular pattern and yellowish areas with subcapsular pinpoint hemorrhages. Edema in body cavities, gallbladder wall, perirenal tissues, initial portion of duodenum, pancreas, and abomasum was also seen. Hemorrhages were also present in subcutaneous tissues, heart, and in mucosal and serosal membranes of the abdominal cavity. The principal histological finding was centrolobular to massive coagulative necrosis associated with centrolobular hemorrhage and congestion and periportal degeneration and tumefaction. There also were lymphoid depletion and necrosis in the germinative centres of lymph nodes, Peyer’s patches, and white pulp of the spleen. Transmission electron microscopy showed necrotic hepatocytes and dilatation in the space of Disse and vascular endothelium. Severe proliferation of smooth endoplasmic reticulum was also seen in animal receiving more than one dosis of larvae. Thawed larvae were desiccated at 100ºC for 24 h and administered to one rabbit that also became ill. Additional two rabbit were dosed with solid or liquid fractions of larvae and only which received the solid fraction died.

A spiny (Palinuridae), slipper (Scyllaridae) and coral (Synaxidae) lobster larval composition and distribution study on the western edge of the Florida Current is presented. From the samples collected during 2007, phyllosoma larvae of Justitia longimanus, Panulirus argus, Parribacus spp., S. americanus, S. depressus and P. gundlachi were positively identified. Relative density catches of the phyllosoma larvae indicate that P. argus (Florida spiny lobster) is the most abundant species in the Straits of Florida. Data gathered during the study shows that P. argus larvae at different developmental stages occur throughout the year which supports the theory of multiple spawning and/or multiple sources. Larvae of genus Panulirus are difficult to distinguish from plankton samples and hence were identified as of P. argus since it is the most common species in the area. Family Palinuridae and Scyllaridae larval density spikes in July and May, coincide with their spawning peak periods of late March – early May and late January to late March respectively. Members of family Palinuridae showed higher densities at night, whereas those of family Scyllaridae showed higher densities during day catches. Larvae of all species showed close association between total length and developmental stage at earlier age. Variability in size and morphology within stage increases with older stages due to multiple molts and/or smaller sample size.

The Florida Keys experienced some of the most drastic transitions from coral to macroalgae dominated states, known as phase-or regime-shifts, of any reefs in the Caribbean. Macroalgae on coral reefs lower coral recruitment by deterring coral settlement either directly through competition or indirectly by changing the chemical environment near the benthos. With evidence of species-specific interactions to coral-macroalgae competition, the type of macroalgae on a phase-shifted coral reef might be more important than just identifying a reef transition. To answer this question, I tested the effect of Laurencia intricata (a macroalgae related to the settlement inducing crustose coralline algae) and Dictyotaceae (known for its toxic or allelopathic compounds) on Porites astreoides planulae behavior, settlement and choice settlement preference, and post-settlement survival. I found that P. astreoides planulae show a positive response to chemical cues released from L. intricata, crustose coralline algae, and species in the Dictyotaceae family. However, the positive chemical cue response becomes algal-specific as larvae start probing for settlement substrate. Providing P. astreoides larvae with a choice between settlement substrates, revealed that the algal structure caused higher settlement next to L. intricata, while Dictyotaceae deterred larval settlement. It may be beneficial for larvae to settle next to L. intricata over Dictyotaceae algae. I identified that post-settlement survival was enhanced when P. astreoides larvae settled next to L. intricata while Dictyotaceae species did not enhance or deter post-settlement survival. These results indicate that coral larvae may be responding differently to a variety of chemical cues. Any chemical or physical cue from a reef may be used by coral larvae to identify and locate settlement substrate on a reef. Once they identify a reef’s location, they express a more selective behavior during settlement by avoiding Dictyotaceae macroalgae and favoring L. intricata. This suggests that the composition of a phase-shifted reef matters to coral recovery, not only that it has shifted to a dominated macroalgal state.

Comprendre la dynamique des populations marines est essentiel à une gestion efficaceet requiert des connaissances sur la dispersion et la connectivité entre populationsqui sont encore très lacunaires. Beaucoup d’organismes marins ont un cycle de viebipartite avec une phase larvaire pélagique qui représente souvent la seule possibilitéde dispersion. De nouvelles techniques de mesure de la dispersion larvaire, parmarquage ou modélisation, ont été développées durant ces quinze dernières années.Cependant, les résultats de ces deux types d’approches ont rarement été comparésau sein d’un même système marin, limitant l’utilisation des modèles de dispersiondans les modèles de métapopulation. Dans cette thèse, nous utilisons ces deux typesd’approches pour étudier la connectivité larvaire d’un poisson de récif corallien,Dascyllus aruanus, dans le lagon sud-ouest de Nouvelle-Calédonie. Notre modèle dedispersion montre que la rétention larvaire présente une variabilité temporelle élevéeà l’échelle lagonaire et à l’échelle d’un patch de récif, et atteint périodiquement desvaleurs élevées malgré des temps moyens de résidence courts. Le marquage artificieltransgénérationnel des otolithes montre des taux d’auto-recrutement relativementbas à l’échelle de la saison reproductive, suggérant une ouverture importante despopulations, et une variabilité temporelle considérable de l’auto-recrutement. Enfin,les grandes différences entre les résultats du modèle et ceux des marquages appuientle besoin de mieux comprendre les processus qui facilitent la rétention larvaire commeles comportements de homing et la circulation des courants à très petite échelle. / Understanding marine populations dynamics is critical to their effective management,and requires information on patterns of dispersal and connectivity that are still poorlyknown. Many marine organisms have a bipartite life history with a pelagic larvalstage that often represents the only opportunity for dispersal. In the last decade,new empirical and simulation approaches to measuring larval dispersal have beendeveloped, but results from these two different approaches have rarely been comparedin the context of a single marine system, impeding the use of larval dispersal modelsin metapopulation models supporting decision making. In this doctoral research, weused both approaches to investigate larval connectivity for a coral reef fish, Dascyllusaruanus, in the South-West Lagoon of New Caledonia. Our biophysical dispersalmodel shows that larval retention exhibits considerable temporal variability at bothlagoon and patch reef scales and periodically reaches large values despite low averagewater residence time. Artificial transgenerational marking of embryonic otoliths inthe wild also showed relatively low self-recruitment rates indicating high populationopenness at the reproductive season scale, with considerable monthly variability ofself-recruitment. Large quantitative discrepancies between simulations and empiricalresults emphasize the need to better understand processes that facilitate local retention,such as homing behavior and very small scale circulation patterns.

The global stressors of ocean warming and acidification, as well as local stressors such as eutrophication, overfishing, and coastal construction, have all contributed to the severe decline in coral populations worldwide. Recovery of coral reefs depends partly on recruitment, which relies on the response of larvae to settlement cues indicative of habitat quality; however, it remains unclear whether recruitment in disturbed areas will be compromised. Specifically, as reefs become more disturbed and dominated by macroalgae, it is important to understand larval behavior in response to changes in habitat quality. In this study, we first assessed the settlement success of newly released Agaricia agariciteslarvae in response to several settlement cues, including temperature and water movement. Then, to test the Desperate Larva Hypothesis, the ability of larvae of different ages (0-7 days) to settle and discriminate between inducing and inhibitory settlement cues was assessed. Newly released larvae displayed a stronger preference for settling on crustose coralline algae (93%) than on ceramic plates in macroalgae- (30%) or conspecific-treated seawater (5%), or filtered seawater (13%) (control). Older larvae became progressively less discriminatory of settlement cues, settling even in response to inhibitory cues. This confirms that, although the absence of good settlement cues initially deters settlement, larvae become desperate as they become older and settle even on unfavorable substrates, thus contributing to recruitment in poor quality habitats.

No description available.

No description available.