Mercury in Florida Bay fish: spatial distribution of elevated concentrations and possible linkages to Everglades restoration

Health advisories are now posted in northern Florida Bay, adjacent to the Everglades, warning of high mercury concentrations in some species of gamefish. Highest concentrations of mercury in both forage fish and gamefish have been measured in the northeastern corner of Florida Bay, adjacent to the dominant freshwater inflows from the Everglades. Thirty percent of spotted seatrout (Cynoscion nebulosus Cuvier, 1830) analyzed exceeded Florida’s no consumption level of 1.5 μg g−1 mercury in this area. We hypothesized that freshwater draining the Everglades served as the major source of methylmercury entering the food web supporting gamefish. A lack of correlation between mercury concentrations and salinity did not support this hypothesis, although enhanced bioavailability of methylmercury is possible as freshwater is diluted with estuarine water. Stable isotopes of carbon, nitrogen, and sulfur were measured in fish to elucidate the shared pathways of methylmercury and nutrient elements through the food web. These data support a benthic source of both methylmercury and nutrient elements to gamefish within the eastern bay, as opposed to a dominant watershed source. Ecological characteristics of the eastern bay, including active redox cycling in near-surface sediments without excessive sulfide production are hypothesized to promote methylmercury formation and bioaccumulation in the benthos. Methylmercury may then accumulate in gamefish through a food web supported by benthic microalgae, detritus, pink shrimp (Farfantepenaeus duorarum Burkenroad, 1939), and other epibenthic feeders. Uncertainty remains as to the relative importance of watershed imports of methylmercury from the Everglades and in situ production in the bay, an uncertainty that needs resolution if the effects of Everglades restoration on mercury levels in fish are to be modeled and managed.

Estimating the Energy Density of Fish: The Importance of Ontogeny

Ontogenetic patterns in the percent dry weight (%DW) and energy density (joules per gram of wet weight) were studied in the early life stages of the subtropical estuarine and marine gray snapper Lutjanus griseus and the warmtemperate estuarine and marine spotted seatrout Cynoscion nebulosus. The %DW was variable for individuals of both species but increased significantly through larval to juvenile stages (<20% for fish ,50 mm standard length to 20–30% for fish >50 mm). The lipid percentage, which was determined only for gray snapper, was also variable between individuals but showed significant increase with body size. Strong relationships between percent dry weight and energy density were evident for both species; however, the slopes of regressions were significantly lower than in general multispecies models, demonstrating the need for species- and stagespecific energy density data in bioenergetics models.

Distribution and abundance of ichthyoplankton in Florida Bay

Ichthyoplankton was sampled at 14 stations with 60 cm bongo nets fitted with 0.333 mm mesh in basins throughout Florida Bay in 1994-1995. In addition, I compared collections made using an epibenthic sled to those made with standard ichthyoplankton bongo nets at four stations during July 1997-November,1999 to determine ifthe two types of gear are complementary. In 1994-1995, in descending order of abundance, Clupeiformes, Gobiidae, Callionymidae, Sciaenidae, Labrisomidae, Soleidae and Blenniidae dominated the ichthyoplankton. Densities of clupeiforms were generally very high (> 100 larvae 100 m-3) or high (10.0 - 99.9 larvae 100 m-3). Gobiid larvae were ubiquitous with highest densities occurring in waters in close proximity to the Gulf of Mexico (109.7 larvae 100 m-3), lowest in two ofthree eastern Florida Bay stations (<1.0 larva 100 m-3). Spotted seatrout, Cynoscion nebulosus, dominated larval sciaenid collections and the only other sciaenid identified to species was the sand seatrout, Cynoscion arenarius. Taxa differed markedly between collections taken by epibenthic sled and standard ichthyoplankton bongo nets. Taxa collected with standard ichthyoplankton gear were those that spawn in Florida Bay and have pelagic larvae (i.e., engraulids and gobiids). Taxa collected with the sled were small resident species that have benthic larvae (i.e., syngnathids and cyprinodonts) or taxa that spawn outside the bay, but use the bay as a nursery area (i.e., gerreids and haemulids). Recently-settled red drum, Sciaenops ocellatus, were collected with the epibenthic sled in November 1999, although juveniles of this important gamefish are rare in the bay.

Report on the Alternative Platform Observer Program in North Carolina: March 2006 to March 2007

In February 2006, an Alternative Platform Observer Program (APP) was implemented in North Carolina (NC) to observe commercial gillnet trips by small vessels [<24 ft (7.2 m)] in nearshore waters out to three nm (5.6 km). Efforts began with outreach to the fishing industry while simultaneously gathering information to be incorporated in a Database of Fishermen. From 30 March 2006 through 31 March 2007, 36 trips were observed. Observed trips of the NC nearshore gillnet fishery targeted seven species: kingfish (Menticirrhus spp.), Spanish mackerel (Scomberomorus maculatus), spiny dogfish (Squalus acanthias), spot (Leiostomus xanthurus), spotted seatrout (Cynoscion nebulosus), striped bass (Morone saxatilis), and weakfish (Cynoscion regalis). Of the 36 trips, 20 (55.6%) were with vessels that were new to the Northeast Fisheries Observer Program (NEFOP), having never carried an observer. Based on the landings data for small vessels from North Carolina Division of Marine Fisheries (NCDMF), the APP has achieved 10.1% coverage by number of trips and 4.0% by pounds landed. No incidental takes of bottlenose dolphins were observed by the APP, although bottlenose dolphins were sighted during 19 (52.8%) observed trips. The APP has drastically increased the number of observed trips of small vessels in the nearshore waters of NC. When combined with trips observed by NEFOP (n=205), the APP resulted in a 15.6% increase in the number of observed gillnet trips. (PDF contains 34 pages)

Results of a fish health survey of North Biscayne Bay, June 1976-June 1977

Fish were collected weekly in Biscayne Bay using a monofilament gill net set from a small skiff during 20-30 minute intervals. Although weekly sampling took place for 2.5 years, only the data from samples collected from June 1976 to June 1977 were used in this document. Abnormal external conditions of fins and body were observed on each fish and recorded. Fish were returned immediately to their habitats. Fish collected in the time period for this study numbered 3,765 and included 32 species. Of these, 16 species, totaling 3,556 fish, were caught in sufficient numbers (20 or more) to warrant data analysis. Only 3 of the 16 species could be considered relatively unafflicted: Aetobatus narinari (spotted eagle ray), Diodon hystrix (porcupinefish), and Selene vomer (lookdown). More than 80% of the examined specimens of these three species were unaffected. Less than 20% of the specimens of Diapterus plumieri (striped mojarra), Micropogonias undulatus (Atlantic croaker), and Pogonias cromis (black drum) displayed normal conditions. The three most afflicted species were Diapterus plumieri, striped mojarra; Micropogonias undulatus, Atlantic croaker; and Pogonias cromis, black drum. Only 7, 3, and 7% respectively showed no external evidence of disease. Data described in this document were originally tabulated in the mid-1970s, remained unpublished, and are no longer available. This document was based on archived unpublished text, a data summary table, and figures. Most of the text and cited references were the ones used in the original manuscript and no attempt was made to update them. (PDF contains 44 pages)

Radiotherapy plus nimotuzumab or placebo in the treatment of high grade glioma patients: results from a randomized, double blind trial

Background The prognosis of patients bearing high grade glioma remains dismal. Epidermal Growth Factor Receptor (EGFR) is well validated as a primary contributor of glioma initiation and progression. Nimotuzumab is a humanized monoclonal antibody that recognizes the EGFR extracellular domain and reaches Central Nervous System tumors, in nonclinical and clinical setting. While it has similar activity when compared to other anti-EGFR antibodies, it does not induce skin toxicity or hypomagnesemia. Methods A randomized, double blind, multicentric clinical trial was conducted in high grade glioma patients (41 anaplastic astrocytoma and 29 glioblastoma multiforme) that received radiotherapy plus nimotuzumab or placebo. Treatment and placebo groups were well-balanced for the most important prognostic variables. Patients received 6 weekly doses of 200 mg nimotuzumab or placebo together with irradiation as induction therapy. Maintenance treatment was given for 1 year with subsequent doses administered every 3 weeks. The objectives of this study were to assess the comparative overall survival, progression free survival, response rate, immunogenicity and safety. Results The median cumulative dose was 3200 mg of nimotuzumab given over a median number of 16 doses. The combination of nimotuzumab and RT was well-tolerated. The most prevalent related adverse reactions included nausea, fever, tremors, anorexia and hepatic test alteration. No anti-idiotypic response was detected, confirming the antibody low immunogenicity. The mean and median survival time for subjects treated with nimotuzumab was 31.06 and 17.76 vs. 21.07 and 12.63 months for the control group. Conclusions In this randomized trial, nimotuzumab showed an excellent safety profile and significant survival benefit in combination with irradiation.

Abundance and distribution patters of Hawaiian odontocetes: focus on O'ahu

This dissertation is an assessment of the status of odontocetes in Hawaiian waters focussing on O´ahu. The work builds on available literature, and on data collected by the author and by others in Hawaiian waters. Abundance and distribution patterns of odontocetes were derived from stranding and aerial survey data. A stranding network operated by the National Marine Fisheries Service, Pacific Area Office collected 187 stranding reports throughout the main Hawaiian Islands between 1937 and 2002. These reports included 16 odontocete species. Number of stranding reports increased over time and was highest on O´ahu. Strandings occurred throughout the year. The difference in number of strandings per month was not significant. Fifteen of the 16 species reported in the stranding record for the main Hawaiian Islands were also reported by aerial survey studies of the area between 1993 and 1998. Only 7 of the species reported were detected during aerial transects around O′ahu between 1998 and 2000. Based on the stranding record, Kogia sp., melon-headed whales, striped dolphins and dwarf killer whale appear to be more common than suggested by aerial surveys. Conversely, pilot whales and bottlenose dolphins were more common, according to aerial surveys, than predicted by the stranding data. Aerial surveys of waters between 0 and 500m around the Island of O′ahu showed that the most abundant species by frequency of occurrence was the pilot whale (30% of sightings), followed by the spinner (16%) and bottlenose dolphin (14%). Because of small sample size, abundance estimates for odontocetes have a high level of uncertainty. The unavailability of a correction factor for g(0)<1, and the reduced visibility below the aircraft further reduced accuracy and increased the inherent underestimation in the data. The most abundant species according to distance sampling estimates were spotted dolphins, pilot whales, false killer whales and spinner dolphins. A natural factor shaping the ecology of odontocete populations is predation pressure both by other odontocetes and, more frequently, by sharks. An account of predation by a tiger shark on a spotted dolphin near Penguin Banks is used as an example of the potential mechanisms of predation by sharks on odontocetes.

Stocks of Dolphins (Stenella spp. and Delphinus delphis) in the Eastern Tropical Pacific: A Phylogeographic Classification

Current information is reviewed that provides clues to the intraspecific structure of dolphin species incidently killed in the yellowfin tuna purse-seine fishery of the eastern tropical Pacific (ETP). Current law requires that management efforts are focused on the intraspecific level, attempting to preserve local and presumably locally adapted populations. Four species are reviewed: pantropical spotted, Stenella attenuata; spinner, S. longirostTis; striped, S. coeruleoalba; and common, Delphinus delphis, dolphins. For each species, distributional, demographic, phenotypic, and genotypic data are summarized, and the putative stocks are categorized based on four hierarchal phylogeographic criteria relative to their probability of being evolutionarily significant units. For spotted dolphins, the morphological similarity of animals from the south and the west argues that stock designations (and boundaries) be changed from the current northern offshore and southern offshore to northeastern offshore and a combined western and southern offshore. For the striped dolphin, we find little reason to continue the present division into geographical stocks. For common dolphins, we reiterate an earlier recommendation that the long-beaked form (Baja neritic) and the northern short-beaked form be managed separately; recent morphological and genetic work provides evidence that they are probably separate species. Finally, we note that the stock structure of ETP spinner dolphins is complex, with the whitebelly form exhibiting characteristics of a hybrid swarm between the eastern and pantropical subspecies. There is little morphological basis at present for division of the whitebelly spinner dolphin into northern and southern stocks. However, we recommend continued separate management of the pooled whitebelly forms, despite their hybrid/intergrade status. Steps should be taken to ensure that management practices do not reduce the abundance of eastern relative to whitebelly spinner dolphins. To do so may lead to increased invasion of the eastern's stock range and possible replacement of the eastern spinner dolphin genome.(PDF file contains 24 pages.)

Distribución de "coxiella burnetii" en los rumiantes domésticos y en la fauna silvestre de la Comunidad Atutónoma Vasca. Evaluacióndel efecto del tratamiento antibiótico y de la vacunación en el control de la fiebre Q en rebaños ovinos naturalmente infectados

243 p. : il.

Some potential occupational and environmental hazards associated with fish pond production in Nigeria

The paper highlights the concept of information and the significance of environmental and occupational hazards associated with pond fish production in Nigeria and discuss the possible options for the ways forward. The major raw material used in fish production system is the organic manure (cow dung, poultry droppings, porcine manure etc) that serves as substrate for heterotrophic production of bacteria and protozoa, which act as food for zooplankton and the fish. The pathogenic organisms (viruses, bacteria, protozoa's, and parasites), are noted for the potential hazard to the fish handlers and consumers. Nine species from seven genera of bacteria associated with fish diseases are found to have association with diseases of human such as typhoid fever, bacillary dysentery and other gastrointestinal tract related problems. Also the environmental contaminants in pond fish production become important because of its significance to consumers' acceptance of the fish products

Engineered underdominance as a method of insect population replacement and reproductive isolation

Insect vector-borne diseases, such as malaria and dengue fever (both spread by mosquito vectors), continue to significantly impact health worldwide, despite the efforts put forth to eradicate them. Suppression strategies utilizing genetically modified disease-refractory insects have surfaced as an attractive means of disease control, and progress has been made on engineering disease-resistant insect vectors. However, laboratory-engineered disease refractory genes would probably not spread in the wild, and would most likely need to be linked to a gene drive system in order to proliferate in native insect populations. Underdominant systems like translocations and engineered underdominance have been proposed as potential mechanisms for spreading disease refractory genes. Not only do these threshold-dependent systems have certain advantages over other potential gene drive mechanisms, such as localization of gene drive and removability, extreme engineered underdominance can also be used to bring about reproductive isolation, which may be of interest in controlling the spread of GMO crops. Proof-of-principle establishment of such drive mechanisms in a well-understood and studied insect, such as Drosophila melanogaster, is essential before more applied systems can be developed for the less characterized vector species of interest, such as mosquitoes. This work details the development of several distinct types of engineered underdominance and of translocations in Drosophila, including ones capable of bringing about reproductive isolation and population replacement, as a proof of concept study that can inform efforts to construct such systems in insect disease vectors.

Proteolytic processing of the nonstructural proteins of dengue 2 virus

The genomes of many positive stranded RNA viruses and of all retroviruses are translated as large polyproteins which are proteolytically processed by cellular and viral proteases. Viral proteases are structurally related to two families of cellular proteases, the pepsin-like and trypsin-like proteases. This thesis describes the proteolytic processing of several nonstructural proteins of dengue 2 virus, a representative member of the Flaviviridae, and describes methods for transcribing full-length genomic RNA of dengue 2 virus. Chapter 1 describes the in vitro processing of the nonstructural proteins NS2A, NS2B and NS3. Chapter 2 describes a system that allows identification of residues within the protease that are directly or indirectly involved with substrate recognition. Chapter 3 describes methods to produce genome length dengue 2 RNA from cDNA templates.

The nonstructural protein NS3 is structurally related to viral trypsinlike proteases from the alpha-, picorna-, poty-, and pestiviruses. The hypothesis that the flavivirus nonstructural protein NS3 is a viral proteinase that generates the termini of several nonstructural proteins was tested using an efficient in vitro expression system and antisera specific for the nonstructural proteins NS2B and NS3. A series of cDNA constructs was transcribed using T7 RNA polymerase and the RNA translated in reticulocyte lysates. Proteolytic processing occurred in vitro to generate NS2B and NS3. The amino termini of NS2B and NS3 produced in vitro were found to be the same as the termini of NS2B and NS3 isolated from infected cells. Deletion analysis of cDNA constructs localized the protease domain necessary and sufficient for correct cleavage to the first 184 amino acids of NS3. Kinetic analysis of processing events in vitro and experiments to examine the sensitivity of processing to dilution suggested that an intramolecular cleavage between NS2A and NS2B preceded an intramolecular cleavage between NS2B and NS3. The data from these expression experiments confirm that NS3 is the viral proteinase responsible for cleavage events generating the amino termini of NS2B and NS3 and presumably for cleavages generating the termini of NS4A and NS5 as well.

Biochemical and genetic experiments using viral proteinases have defined the sequence requirements for cleavage site recognition, but have not identified residues within proteinases that interact with substrates. A biochemical assay was developed that could identify residues which were important for substrate recognition. Chimeric proteases between yellow fever and dengue 2 were constructed that allowed mapping of regions involved in substrate recognition, and site directed mutagenesis was used to modulate processing efficiency.

Expression in vitro revealed that the dengue protease domain efficiently processes the yellow fever polyprotein between NS2A and NS2B and between NS2B and NS3, but that the reciprocal construct is inactive. The dengue protease processes yellow fever cleavage sites more efficiently than dengue cleavage sites, suggesting that suboptimal cleavage efficiency may be used to increase levels of processing intermediates in vivo. By mutagenizing the putative substrate binding pocket it was possible to change the substrate specificity of the yellow fever protease; changing a minimum of three amino acids in the yellow fever protease enabled it to recognize dengue cleavage sites. This system allows identification of residues which are directly or indirectly involved with enzyme-substrate interaction, does not require a crystal structure, and can define the substrate preferences of individual members of a viral proteinase family.

Full-length cDNA clones, from which infectious RNA can be transcribed, have been developed for a number of positive strand RNA viruses, including the flavivirus type virus, yellow fever. The technology necessary to transcribe genomic RNA of dengue 2 virus was developed in order to better understand the molecular biology of the dengue subgroup. A 5' structural region clone was engineered to transcribe authentic dengue RNA that contains an additional 1 or 2 residues at the 5' end. A 3' nonstructural region clone was engineered to allow production of run off transcripts, and to allow directional ligation with the 5' structural region clone. In vitro ligation and transcription produces full-length genomic RNA which is noninfectious when transfected into mammalian tissue culture cells. Alternative methods for constructing cDNA clones and recovering live dengue virus are discussed.

Características clínicas e frequência de diarreia por norovírus em crianças hospitalizadas, vacinadas e não vacinadas contra rotavírus Rio de Janeiro, Brasil, 2004-2009

Os norovírus (NV) são uma importante causa de hospitalização infantil. Crianças internadas por gastroenterite por NV (GENV) são consideradas portadoras de diarreia grave. O objetivo desse estudo, realizado na cidade do Rio de Janeiro, Brasil, é descrever as características clínicas e a frequência da diarreia por NV em crianças hospitalizadas, comparando as taxas de detecção de NV em crianças vacinadas e não vacinadas contra rotavírus (Rotarix). Foram coletadas 659 amostras de fezes de igual número de crianças e encaminhadas para análise pela reação em cadeia pela polimerase, precedida de transcrição reversa no período de janeiro de 2004 a dezembro de 2009. O percentual de amostras positivas para os NV foi de 27,3% nesse período. Das 180 amostras positivas para NV, 55% tiveram origem na comunidade (aqCo) e 45% foram de aquisição nosocomial (aqNo). O percentual de GENV nos dois anos anteriores (2004 e 2005) à introdução da vacina Rotarix foi de 28,3%, sendo 11,3% o percentual de amostras aqCo. Nos dois anos posteriores (2008 e 2009), a GENV significou 24,4%, e as amostras aqCo foram 14,9% (p<0,05). Em 647 crianças, 494 não receberam a vacina Rotarix, enquanto 151 crianças receberam, pelo menos, uma dose. O percentual de GENV foi de 23,8% e 39,7%, respectivamente (p<0,05). Apesar do comportamento sazonal dos casos de GENV aqCo, esse fato não teve significância estatística. Das 180 crianças, 61,6% tinham peso ≤ p10 do NCHS, 82,2% tinham idade ≤ 5anos. As crianças com idade ≤ 2 anos foram mais acometidas nos casos de aqCo do que àquelas de aqNo (p<0,05). Foram observados em 82 crianças: vômitos (73,2%), febre (54,9%), tosse (20,7%), coriza (2,2%), sangue nas fezes (8,5%), erupção cutânea (4,9%) e broncoespasmo (7,3%). Houve significância estatística com relação à frequência maior de febre, coriza, tosse e broncoespasmo nas crianças com GENV de aqCo do que naquelas de aqNo (p<0,05). De 69 crianças, 73,9% apresentaram desidratação e, dessas, 76,5% necessitaram de hidratação venosa. Esses dados tiveram significância estatística, representada por maiores percentuais nas crianças com GENV de aqCo do que naquelas de aqNo (p<0,05). Esse estudo demonstra que os NV foram um importante agente etiológico nos casos de gastroenterites em crianças hospitalizadas e responsável por altas taxas de infecções nosocomiais. Estatisticamente, não foi comprovada uma tendência de aumento dos casos de GENV no período do estudo, como também do aumento da frequência de GENV nos anos posteriores em relação aos anos anteriores à introdução da vacina Rotarix no Brasil em 2006. No entanto, houve significância estatística quando foi avaliado o percentual de GENV em crianças hospitalizadas vacinadas e não vacinadas contra RV. Um aumento dos casos de GENV em crianças poderá vir a acontecer nos próximos anos, quando é esperado que um número maior de crianças será vacinado contra RV. Tosse, coriza e broncoespasmo são sintomas que devem ser mais detalhadamente investigados. Estratégias de prevenção contra a disseminação dos NV são condutas importantes em unidades de internação. Uma vacina eficaz contra norovírus pode ser um benefício significativo para reduzir o percentual de crianças hospitalizadas por diarreia.

Viral metagenomic analysis of feces of wild small carnivores

Background: Recent studies have clearly demonstrated the enormous virus diversity that exists among wild animals. This exemplifies the required expansion of our knowledge of the virus diversity present in wildlife, as well as the potential transmission of these viruses to domestic animals or humans. Methods: In the present study we evaluated the viral diversity of fecal samples (n = 42) collected from 10 different species of wild small carnivores inhabiting the northern part of Spain using random PCR in combination with next-generation sequencing. Samples were collected from American mink (Neovison vison), European mink (Mustela lutreola), European polecat (Mustela putorius), European pine marten (Martes martes), stone marten (Martes foina), Eurasian otter (Lutra lutra) and Eurasian badger (Meles meles) of the family of Mustelidae; common genet (Genetta genetta) of the family of Viverridae; red fox (Vulpes vulpes) of the family of Canidae and European wild cat (Felis silvestris) of the family of Felidae. Results: A number of sequences of possible novel viruses or virus variants were detected, including a theilovirus, phleboviruses, an amdovirus, a kobuvirus and picobirnaviruses. Conclusions: Using random PCR in combination with next generation sequencing, sequences of various novel viruses or virus variants were detected in fecal samples collected from Spanish carnivores. Detected novel viruses highlight the viral diversity that is present in fecal material of wild carnivores.

Comparative feeding ecology of two elasmobranch species, Squalus blainville and Scyliorhinus canicula, off the coast of Portugal

The small-spotted catshark (Scyliorhinus canicula) (Linnaeus, 1758) and the longnose spurdog (Squalus blainville) (Risso, 1826) are two species occurring in the European and western African continental shelves with a wide geographical distribution. In this study, the diet of S. blainville and S. canicula off the Portuguese western Atlantic coast was investigated in 2006 by collecting monthly samples of these two species from local fishing vessels. In the stomachs of both species, crustaceans and teleosts were the dominant prey items, and molluscs, polychaetes, echinoderms, and sipunculids were found in lower abundance. In S. canicula, urochordate and chondrichthyan species were also observed in stomachs and were classified as accidental prey items. Scyliorhinus canicula consumed a broader group of prey items than did S. blainville. A significant diet overlap was observed, despite both species occupying different depth ranges over the continental shelf. Scyliorhinus canicula exhibited a consistency in diet composition among seasons, sexes, and maturity stages. Nonetheless, for both adults and juveniles, an increase in relative abundance of teleosts in the diet was observed in the spring and summer. This study provides evidence of the importance of S. canicula and S. blainville as benthic and pelagic predators along the western Atlantic coast.