ExoU, uma proteína do sistema de secreção do tipo III de Pseudomonas aeruginosa, induz a secreção de eicosanóides por células epiteliais respiratórias. Papel dos corpúsculos lipídicos neste processo

Para pesquisar o papel de ExoU no desencadeamento de resposta inflamatória nas vias aéreas, células epiteliais respiratórias humanas (CERs) da linhagem BEAS-2B foram tratadas com AA radiomarcado e infectadas com a cepa PA103 de P. aeruginosa, que secreta ExoU, e com os mutantes PA103exoU (com deleção do gene exoU), PA103ΔUT/exoU (com deleção de exoU e complementação com o gene funcional) e PA103UT/S142A (com deleção de exoU e complementação com gene com mutagênese sítio-específica no domínio catalítico da enzima). Após 1 hora, a liberação de AA pelas culturas infectadas com as cepas produtoras de ExoU foi significativamente superior à observada em culturas infectadas pelas cepas não-produtoras ou por células controle. O tratamento das bactérias com MAFP, um inibidor de PLA2, resultou em significativa redução na liberação de AA. Células infectadas pelas cepas PA103 e PA103ΔUT/exoU secretaram PGE2 e LTB4 em concentrações significativamente maiores que as secretadas por células infectadas pelas demais cepas ou não infectadas. O tratamento com o MAFP reduziu significativamente a secreção de PGE2. A análise, por citometria de fluxo, de células infectadas e não infectadas tratadas com anticorpo anti-COX-2 mostrou que o percentual de células infectadas por PA103 marcadas foi significativamente superior ao percentual encontrado em culturas controle. Nenhuma diferença foi observada quanto ao percentual de células marcadas em culturas infectadas por PA103ΔexoU. O tratamento das culturas com NS-398 (um inibidor seletivo de COX-2) resultou na diminuição significativa da concentração de PGE2, secretada por células infectadas com PA103, mas não por células infectadas com PA103ΔexoU ou por células controle. Corpúsculos lipídicos (CLs) são domínios citoplasmáticos ricos em COX-2 e outras enzimas responsáveis pelo metabolismo do AA, sede da produção de eicosanóides. Como células infectadas pelas cepas produtoras de ExoU liberam AA livre, formulamos a hipótese de que a maior produção de eicosanóides por estas células seria dependente da indução do aumento no número dos CLs. No entanto, a análise por citometria de fluxo de células tratadas com uma sonda lipofílica com afinidade com os CLs mostrou que o percentual de células marcadas em culturas infectadas pelas cepas produtoras de ExoU foi significativamente inferior ao percentual em culturas controle ou infectadas pelas outras 2 cepas bacterianas. O tratamento das células com MAFP inibiu significativamente a redução do percentual de células contendo CLs. A análise, por citometria de fluxo, de células controle ou infectadas tratadas simultaneamente com a sonda lipofílica e com o anticorpo anti-PGE2, mostrou, em células infectadas com PA103, a redução da mediana da intensidade de marcação com a sonda lipofílica e o aumento da mediana da intensidade de marcação com o anticorpo anti-PGE2. Nossa hipótese é que a presença de ExoU nas células infectadas com a cepa PA103 resulte no metabolismo de glicerofosfolipídios presente nos CLs levando à diminuição da afinidade dos CLs pela sonda lipofílica e à síntese local de PGE2.

Food habits of Sowerby's beaked whales (Mesoplodon bidens) taken in the pelagic drift gillnet fishery of the western North Atlantic

We describe the food habits of the Sowerby’s beaked whale (Mesoplodon bidens) from observations of 10 individuals taken as bycatch in the pelagic drift gillnet fishery for Swordfish (Xiphias gladius) in the western North Atlantic and 1 stranded individual from Kennebunk, Maine. The stomachs of 8 bycaught whales were intact and contained prey. The diet of these 8 whales was dominated by meso- and benthopelagic fishes that composed 98.5% of the prey items found in their stomachs and cephalopods that accounted for only 1.5% of the number of prey. Otoliths and jaws representing at least 31 fish taxa from 15 families were present in the stomach contents. Fishes, primarily from the families Moridae (37.9% of prey), Myctophidae (22.9%), Macrouridae (11.2%), and Phycidae (7.2%), were present in all 8 stomachs. Most prey were from 5 fish taxa: Shortbeard Codling (Laemonema barbatulum) accounted for 35.3% of otoliths, Cocco’s Lanternfish (Lobianchia gemellarii) contributed 12.9%, Marlin-spike (Nezumia bairdii) composed 10.8%, lanternfishes (Lampanyctus spp.) accounted for 8.4%; and Longfin Hake (Phycis chesteri) contributed 6.7%. The mean number of otoliths per stomach was 1196 (range: 327–3452). Most of the fish prey found in the stomachs was quite small, ranging in length from 4.0 to 27.7 cm. We conclude that the Sowerby’s beaked whales that we examined in this study fed on large numbers of relatively small meso- and benthopelagic fishes that are abundant along the slope and shelf break of the western North Atlantic.

Strandings as indicators of marine mammal biodiversity and human interactions off the coast of North Carolina

The adjacency of 2 marine biogeographic regions off Cape Hatteras, North Carolina (NC), and the proximity of the Gulf Stream result in a high biodiversity of species from northern and southern provinces and from coastal and pelagic habitats. We examined spatiotemporal patterns of marine mammal strandings and evidence of human interaction for these strandings along NC shorelines and evaluated whether the spatiotemporal patterns and species diversity of the stranded animals reflected published records of populations in NC waters. During the period of 1997–2008, 1847 stranded animals were documented from 1777 reported events. These animals represented 9 families and 34 species that ranged from tropical delphinids to pagophilic seals. This biodiversity is higher than levels observed in other regions. Most strandings were of coastal bottlenose dolphins (Tursiops truncatus) (56%), harbor porpoises (Phocoena phocoena) (14%), and harbor seals (Phoca vitulina) (4%). Overall, strandings of northern species peaked in spring. Bottlenose dolphin strandings peaked in spring and fall. Almost half of the strandings, including southern delphinids, occurred north of Cape Hatteras, on only 30% of NC’s coastline. Most stranded animals that were positive for human interaction showed evidence of having been entangled in fishing gear, particularly bottlenose dolphins, harbor porpoises, short-finned pilot whales (Globicephala macrorhynchus), harbor seals, and humpback whales (Megaptera novaeangliae). Spatiotemporal patterns of bottlenose dolphin strandings were similar to ocean gillnet fishing effort. Biodiversity of the animals stranded on the beaches reflected biodiversity in the waters off NC, albeit not always proportional to the relative abundance of species (e.g., Kogia species). Changes in the spatiotemporal patterns of strandings can serve as indicators of underlying changes due to anthropogenic or naturally occurring events in the source populations.

Temporal and spatial aspects of bottlenose dolphin occurrence in coastal and estuarine waters near Charleston, South Carolina

The spatial and temporal occurrence of Atlantic bottlenose dolphins (Tursiops truncatus) in the coastal and estuarine waters near Charleston, SC were evaluated. Sighting and photographic data from photo-identification (ID), remote biopsy, capture-release and radio-tracking studies, conducted from 1994 through 2003, were analyzed in order to further delineate residence patterns of Charleston area bottlenose dolphins. Data from 250 photo-ID, 106 remote biopsy, 15 capture-release and 83 radio-tracking surveys were collected in the Stono River Estuary (n = 247), Charleston Harbor (n = 86), North Edisto River (n = 54), Intracoastal Waterway (n = 26) and the coastal waters north and south of Charleston Harbor (n = 41). Coverage for all survey types was spatially and temporally variable, and in the case of biopsy, capture-release and radio-tracking surveys, data analyzed in this report were collected incidental to other research. Eight-hundred and thirty-nine individuals were photographically identified during the study period. One-hundred and fifteen (13.7%) of the 839 photographically identified individuals were sighted between 11-40 times, evidence of consistent occurrence in the Charleston area (i.e., site fidelity). Adjusted sighting proportions (ASP), which reflect an individual’s sighting frequency in a subarea relative to other subareas after adjusting for survey effort, were analyzed in order to evaluate dolphin spatial occurrence. Forty-three percent (n = 139) of dolphins that qualified for ASP analyses exhibited a strong subarea affiliation while the remaining 57% (n = 187) showed no strong subarea preference. Group size data were derived from field estimates of 2,342 dolphin groups encountered in the five Charleston subareas. Group size appeared positively correlated with degree of “openness” of the body of water where dolphins were encountered; and for sightings along the coast, group size was larger during summer months. This study provides valuable information on the complex nature of bottlenose dolphin spatial and temporal occurrence near Charleston, SC. In addition, it helps us to better understand the stock structure of dolphins along the Atlantic seaboard.

Protocols for conducting dolphin capture-release health assessment studies

Marine mammals, such as dolphins, can serve as key indicator species in coastal areas by reflecting the effects of natural and anthropogenic stressors. As such they are often considered sentinels of environmental and ecosystem health (Bossart 2006; Wells et al. 2004; Fair and Becker 2000). The bottlenose dolphin is an apex predator and a key component of many estuarine environments in the southeastern United States (Woodward-Clyde Consultants 1994; SCDNR 2005). Health assessments of dolphins are especially critical in areas where populations are depleted, show signs of epidemic disease and/or high mortality and/or where habitat is being altered or impacted by human activities. Recent assessments of environmental conditions in the Indian River Lagoon, Florida (IRL) and the estuarine waters surrounding Charleston, South Carolina (CHS) highlight the need for studies of the health of local bottlenose dolphins. While the condition of southeastern estuaries was rated as fair in the National Coastal Condition Report (U.S. EPA 2001), it was noted that the IRL was characterized by poorer than expected benthic communities, significant sediment toxicity and increased nutrient concentrations. Similarly, portions of the CHS estuary have sediment concentrations of aliphatic aromatic hydrocarbons, select inorganic metals, and some persistent pesticides far in excess of reported bioeffect levels (Hyland et al. 1998). Long-term trends in water quality monitoring and recent scientific research suggest that waste load assimilation, non-point source runoff impacts, contaminated sediments, and toxic pollutants are key issues in the CHS estuary system. Several ‘hot spots’ with high levels of heavy metals and organic compounds have been identified (Van Dolah et al. 2004). High concentrations of anthropogenic trace metals, polychlorinated biphenyls (PCB’s) and pesticides have been found in the sediments of Charleston Harbor, as well as the Ashley and Cooper Rivers (Long et al. 1998). Two superfund sites are located within the CHS estuary and the key contaminants of concern associated with these sites are: polycyclic aromatic hydrocarbons (PAH), lead, chromium, copper, arsenic, zinc and dioxin. Concerns related to the overall health of IRL dolphins and dermatologic disease observed in many dolphins in the area (Bossart et al. 2003) initiated an investigation of potential factors which may have impacted dolphin health. From May-August 2001, 35 bottlenose dolphins died in the IRL during an unusual mortality event (MMC 2003). Many of these dolphins were diagnosed with a variety of skin lesions including proliferative ulcerative dermatitis due to protozoa and fungi, dolphin pox and a vesicular dermatopathy of unknown etiology (Bossart et al. 2003). Multiple species from fish to dolphins in the IRL system have exhibited skin lesions of various known and unknown etiologies (Kane et al. 2000; Bossart et al. 2003; Reif et al. 2006). On-going photo-identification (photo-ID) studies have documented skin diseases in IRL dolphins (Mazzoil et al. 2005). In addition, up to 70% of green sea turtles in the IRL exhibit fibropapillomas, with the highest rates of occurrence being seen in turtles from the southern IRL (Hirama 2001).

Harmful algal blooms in coastal waters: options for prevention, control and mitigation

This report is the product of a panel of experts in the science of blooms of unicellular marine algae which can cause mass mortalities in a variety of marine organisms and cause illness and even death in humans who consume contaminated seafood. These phenomena are collectively termed harmful algal blooms or HABs for short. As a counterpart to recent assessments of the priorities for scientific research to understand the causes and behavior of HABs, this assessment addressed the management options for reducing their incidence and extent (prevention), actions that can quell or contain blooms (control), and steps to reduce the losses of resources or economic values and minimize human health risks (mitigation). This assessment is limited to an appraisal of scientific understanding, but also reflects consideration of information and perspectives provided by regional experts, agency managers and user constituencies during three regional meetings. The panel convened these meetings during the latter half of 1996 to solicit information and opinions from scientific experts, agency managers and user constituencies in Texas, Washington, and Florida. The panel's assessment limited its attention to those HABs that result in neurotoxic shellfish poisoning, paralytic shellfish poisoning, brown tides, amnesic shellfish poisoning, and aquaculture fish kills. This covers most, but certainly not all, HAB problems in the U.S.

Development, validation, and utilization of a competitive enzyme-linked immunosorbent assay for the detection of antibodies against Brucella species in marine mammals

A competitive enzyme-linked immunosorbent assay (cELISA) was developed by using a whole-cell antigen from a marine Brucella sp. isolated from a harbor seal (Phoca vitulina). The assay was designed to screen sera from multiple marine mammal species for the presence of antibodies against marine-origin Brucella. Based on comparisons with culture-confirmed cases, specificity and sensitivity for cetacean samples tested were 73% and 100%, respectively. For pinniped samples, specificity and sensitivity values were 77% and 67%, respectively. Hawaiian monk seal (Monachus schauinslandi; n = 28) and bottlenose dolphin (Tursiops truncatus; n = 48) serum samples were tested, and the results were compared with several other assays designed to detect Brucella abortus antibodies. The comparison testing revealed the marine-origin cELISA to be more sensitive than the B. abortus tests by the detection of additional positive serum samples. The newly developed cELISA is an effective serologic method for detection of the presence of antibodies against marine-origin Brucella sp. in marine mammals.

Influence of microbial interactions on the susceptibility of Karenia spp. to algicidal bacteria

A bacterial strain (D38BY) belonging to the family Flavobacteriaceae and antagonistic towards an algicidal bacterium (strain S03; Flavobacteriaceae) was isolated from a culture of the red tide dinoflagellate Karenia brevis that had previously been characterized as resistant to attack by strain S03. This antagonistic bacterium increased the survival time of otherwise susceptible, bacteriafree K. brevis cultures in a concentration-dependent manner during exposure to the algicidal bacterium. Experimental evidence indicated that direct contact was required in order for strain D38BY to inhibit the killing activity of algicidal strain S03. While further work is needed to determine its precise mode of action, the antagonistic properties of strain D38BY provide further evidence that the resistance or susceptibility of certain algal taxa to algicidal attack can be more a function of interactions within the ambient microbial community than an intrinsic property of the alga.

Bartonella species detection in captive, stranded and free-ranging cetaceans

We present prevalence of Bartonella spp. for multiple cohorts of wild and captive cetaceans. One hundred and six cetaceans including 86 bottlenose dolphins (71 free-ranging, 14 captive in a facility with a dolphin experiencing debility of unknown origin, 1 stranded), 11 striped dolphins, 4 harbor porpoises, 3 Risso's dolphins, 1 dwarf sperm whale and 1 pygmy sperm whale (all stranded) were sampled. Whole blood (n = 95 live animals) and tissues (n = 15 freshly dead animals) were screened by PCR (n = 106 animals), PCR of enrichment cultures (n = 50 animals), and subcultures (n = 50 animals). Bartonella spp. were detected from 17 cetaceans, including 12 by direct extraction PCR of blood or tissues, 6 by PCR of enrichment cultures, and 4 by subculture isolation. Bartonella spp. were more commonly detected from the captive (6/14, 43%) than from free-ranging (2/71, 2.8%) bottlenose dolphins, and were commonly detected from the stranded animals (9/21, 43%; 3/11 striped dolphins, 3/4 harbor porpoises, 2/3 Risso's dolphins, 1/1 pygmy sperm whale, 0/1 dwarf sperm whale, 0/1 bottlenose dolphin). Sequencing identified a Bartonella spp. most similar to B. henselae San Antonio 2 in eight cases (4 bottlenose dolphins, 2 striped dolphins, 2 harbor porpoises), B. henselae Houston 1 in three cases (2 Risso's dolphins, 1 harbor porpoise), and untyped in six cases (4 bottlenose dolphins, 1 striped dolphin, 1 pygmy sperm whale). Although disease causation has not been established, Bartonella species were detected more commonly from cetaceans that were overtly debilitated or were cohabiting in captivity with a debilitated animal than from free-ranging animals. The detection of Bartonella spp. from cetaceans may be of pathophysiological concern.

Anti-grazing properties of the toxic dinoflagellate Karlodinium veneficum during predator-prey interactions with the copepod Acartia tonsa

Karlodinium veneficum (syn. Karlodinium micrum, Bergholtz et al. 2006; J Phycol 42:170–193) is a small athecate dinoflagellate commonly present in low levels in temperate, coastal waters. Occasionally, K. veneficum forms ichthyotoxic blooms due to the presence of cytotoxic, hemolytic compounds, putatively named karlotoxins. To evaluate the anti-grazing properties of these karlotoxins, we conducted food removal experiments using the cosmopolitan copepod grazer Acartia tonsa. Wild-caught, adult female A. tonsa were exposed to 6 monoalgal or mixed algal diets made using bloom concentrations of toxic (CCMP 2064) and non-toxic (CSIC1) strains of K. veneficum. Ingestion and clearance rates were calculated using the equations of Frost (1972). Exposure to the toxic strain of K. veneficum did not contribute to an increased mortality of the copepods and no significant differences in copepod mortality were found among the experimental diets. However, A. tonsa had significantly greater clearance and ingestion rates when exposed to a monoalgal diet of the non-toxic strain CSIC1 than when exposed to the monoalgal diet of toxic strain CCMP 2064 and mixed diets dominated by this toxic strain. These results support the hypothesis that karlotoxins in certain strains of K. veneficum deter grazing by potential predators and contribute to the formation and continuation of blooms.

Synopsis of researcher meeting -- Bottlenose Dolphin Health & Risk Assessment Project, February 22-24, 2005

A meeting was convened on February 22-24, 2005 in Charleston, South Carolina to bring together researchers collaborating on the Bottlenose Dolphin Health and Risk Assessment (HERA) Project to review and discuss preliminary health-related findings from captured dolphins during 2003 and 2004 in the Indian River Lagoon (IRL), FL and Charleston (CHS), SC. Over 30 researchers with diverse research expertise representing government, academic and marine institutions participated in the 2-1/2 day meeting. The Bottlenose Dolphin HERA Project is a comprehensive, integrated, multi-disciplinary research program designed to assess environmental and anthropogenic stressors, as well as the health and long-term viability of Atlantic bottlenose dolphins (Tursiops truncatus). Standardized and comprehensive protocols are being used to evaluate dolphin health in the coastal ecosystems in the IRL and CHS. The Bottlenose Dolphin Health and Risk Assessment (HERA) Project was initiated in 2003 by Dr. Patricia Fair at the National Oceanic and Atmospheric Administration/National Ocean Service/Center for Coastal Environmental Health and Biomolecular Research and Dr. Gregory Bossart at the Harbor Branch Oceanographic Institution under NMFS Scientific Research Permit No. 998-1678-00 issued to Dr. Bossart. Towards this end, this study focuses on developing tools and techniques to better identify health threats to these dolphins, and to develop links to possible environmental stressors. Thus, the primary objective of the Dolphin HERA Project is to measure the overall health and as well as the potential health hazards for dolphin populations in the two sites by performing screening-level risk assessments using standardized methods. The screening-level assessment involves capture, sampling and release activities during which physical examinations are performed on dolphins and a suite of nonlethal morphologic and clinicopathologic parameters, to be used to develop indices of dolphin health, are collected. Thus far, standardized health assessments have been performed on 155 dolphins during capture-release studies conducted in Years 2003 and 2004 at the two sites. A major collaboration has been established involving numerous individuals and institutions, which provide the project with a broad assessment capability toward accomplishing the goals and objectives of this project.

Some perspectives on the late Quaternary paleoclimate of Beringia

EXTRACT (SEE PDF FOR FULL ABSTRACT): Analyses of the modern summer synoptic climatology of Beringia illustrate that the region cannot be treated as a homogenous climatic unit as a result of different circulation controls that operate over the region. GCM (general circulation model) simulations and information from the modern synoptic climatology were used to infer the summer paleosynoptic climatology of the region since the last glacial maximum. ... Variations in these climatic controls offer important implications in assessing the vegetation histories of western Beringia versus eastern Beringia.

Lista de peces de la provincia de San Luis.

Esta serie tiene como finalidad dar a conocer las especies presentes en los diferentes estados provinciales. Tomando como base los trabajos de López et al. (2003), Reis et al. (2003), y Liotta (2006), mencionamos para cada territorio los cambios y novedades posteriores a estas publicaciones. Consideramos que este modesto aporte contribuirá a precisar el conocimiento ictiofaunístico regional, ya que, además de las listas de especies, adjuntamos bibliografía de referencia y el marco biogeográfico e hídrico correspondientes, que podrán ser de utilidad para quienes hagan uso de este trabajo. Por otra parte entendemos que la participación de autores involucrados en la región considerada, le da un verdadero sentido federal a esta contribución, además de reforzar vínculos en los protagonistas de nuestra especialidad. En este nuevo número presentamos la provincia de San Luis que se encuentra limitada al norte por San Juan y La Rioja, al este por Córdoba, al oeste por Mendoza y al sur por la provincia de La Pampa. Si bien las especies potencialmente pueden ampliar su rango de distribución, ante cambios climáticos y/o modificaciones de cuencas, es importante considerar para futuros análisis, la acción generada por la antropocoria. Esta, muchas veces realizada de forma involuntaria por los propios pescadores, y en otras ocasiones, bajo responsabilidad de los organismos del estado han desvirtuado la distribución natural de varias especies, afectando tanto estudios zoogeográficos como las medidas dirigidas a la conservación y manejo de los recursos. A las 16 especies citadas por Liotta 2006 para la provincia debemos agregar nuevas citas de especies introducidas (Tabla I y II). Además consideramos que tanto Trichomycterus tenuis como Jenynsia obscura requieren ser confirmadas para la provincia ya que sólo se cuenta con un registro (Tabla III).