Occurrence of Methanogenic Archaea in Highly Polluted Sediments of Tropical Santos-So Vicente Estuary (So Paulo, Brazil)

Little is known about the ability of methanogens to grow and produce methane in estuarine environments. In this study, traditional methods for cultivating strictly anaerobic microorganisms were combined with Fluorescence in situ hybridization (FISH) technique to enrich and identify methanogenic Archaea cultures occurring in highly polluted sediments of tropical Santos-So Vicente Estuary (So Paulo, Brazil). Sediment samples were enriched at 30A degrees C under strict anaerobic and halophilic conditions, using a basal medium containing 2% of sodium chloride and amended with glucose, methanol, and sodium salts of acetate, formate and lactate. High methanogenic activity was detected, as evidenced by the biogas containing 11.5 mmol of methane at 20 days of incubation time and methane yield of 0.138-mmol CH(4)/g organic matter/g volatile suspense solids. Cells of methanogenic Archaea were selected by serial dilution in medium amended separately with sodium acetate, sodium formate, or methanol. FISH analysis revealed the presence of Methanobacteriaceae and Methanosarcina sp. cells.

Influence of allochthonous organic matter on bacterioplankton biomass and activity in a eutrophic, sub-tropical estuary

Heterotrophic bacterial and phytoplankton biomass, production, specific growth rates, and growth efficiencies were studied in the Northern region of the Cananeia-Iguape estuarine system, which has recently experienced an intense eutrophication due to anthropogenic causes. Two surveys were carried out during spring and neap tide periods of the dry season of 2005 and the rainy season of 2006. This region receives large freshwater inputs with organic seston and phosphate concentrations that reach as high as 1.0 mg l(-1) and 20.0 mu M, respectively. Strong decreasing gradients of seston and dissolved inorganic nutrients were observed from the river/estuary boundary to the estuary/coastal interface. Gradients were also observed in phytoplankton and bacterial production rates. The production rates of phytoplankton were 5.6-fold higher (mean 8.5 mu g Cl(-1) h(-1)) during the dry season. Primary production rates (PP) positively correlated with salinity and euphoric depth, indicating that phytoplankton productivity was light-limited. On the other hand, bacterial biomass (BB) and production rates (BP) were 1.9- and 3.7-fold higher, respectively, during the rainy season, with mean values of up to 40.4 mu g Cl(-1) and 7.9 mu g Cl(-1) h-1, respectively. Despite such a high BP, bacterial abundance remained <2 x 106 cells ml(-1), indicating that bacterial production and removal were coupled. Mean specific growth rates ranged between 0.9 and 5.5 d(-1). BP was inversely correlated with salinity and positively correlated with temperature, organic matter, exopolymer particles, and particulate-attached bacteria; this last accounted for as much as 89.6% of the total abundance. During the rainy season, BP was generally much higher than PP, and values of BP/PP > 20 were registered during high freshwater input, suggesting that under these conditions, bacterial activity was predominantly supported by allochthonous inputs of organic carbon. In addition, BB probably represented the main pathway for the synthesis of high-quality (low C:N) biomass that may have been available to the heterotrophic components of the plankton food web, particularly nanoheterotrophs. (C) 2008 Elsevier Ltd. All rights reserved.

Bacterial and phytoplankton dynamics in a sub-tropical estuary

Heterotrophic bacterial and biomass, production, specific growth rates and growth efficiencies were studied in July 2001 and January 2002 during both spring and neap tides, along a tidal cycle, at three sites in a subtropical estuary. Major freshwater inputs located in the Northern region led to differences in both and bacterioplankton biomass and activity along the estuary. While in the Northern region is light-limited, with mean production (PP) between 1.1 and 1.9 mu g C l(-1) h(-1) and mean specific growth rates (PSG) between 0.14 and 0.16 d(-1), the Southern region registered values as high as 24.7 mu g C l(-1) h(-1) for PP and 2.45 d(-1) (mean PP between 3.4 and 7.3 mu g C l(-1) h(-1); mean PSG between 0.28 and 0.57 d(-1)). On the other hand, maximum bacterial production (BP: 63.8 mu g C l(-1) h(-1)) and specific growth rate (BSG: 32.26 d(-1)) were observed in the Northern region (mean BP between 3.4 and 12.8 mu g C l(-1) h(-1); mean BSG between 1.98 and 6.67 day(-1)). These bacterial activity rates are among the highest recorded rates in estuarine and coastal waters, indicating that this system can be highly heterotrophic, due to high loads of allochthonous carbon (mainly derived from mangrove forest). Our results also showed that, despite that BP rates usually exceeded PP, in the Southern region BP may be partially supported (similar to 45%) by PP, since a significant regression was observed between BP and PP (r = 0.455, P < 0.001).

Life cycle and reproductive traits of marine podonids (Cladocera, Onychopoda) in a coastal subtropical area

We studied the temporal distribution and reproductive biology of marine podonids during two consecutive years off Ubatuba, southeast coast of Brazil. Podonid specimens and their eggs and embryos were counted, measured and classified into categories. Pseudevadne tergestina was the most abundant species, and was more abundant in surface layers, in warm seasons, when the water column was stratified because of bottom intrusions of the cold and nutrient-rich South Atlantic Central Water (SACW) onto the inner shelf. Evadne spinifera had a similar temporal and vertical distribution, but with lower abundance and frequency. Pleopis schmackeri did not show a clear seasonal distribution, but preferred bottom layers. Pleopis polyphemoides and Podon intermedius occurred in low abundances, and only under SACW influence. Parthenogenetic females were dominant among all podonid species. Gamogenetic females of P. polyphemoides and P. intermedius were observed, but males of neither species occurred. This suggests that in tropical and subtropical regions, P. tergestina, E. spinifera and P. schmackeri reproduce through parthenogenesis during most of the year.

Feeding ecology of the Brazilian silverside Atherinella brasiliensis (Atherinopsidae) in a sub-tropical estuarine ecosystem

The feeding ecology of the Brazilian silverside, Atherinella brasiliensis, in a sub-tropical estuary of Brazil was investigated through the gut analysis of 1431 individuals. We described dietary composition and analysed seasonal, estuarine habitat, and body size variations in the diet; trophic level; feeding diversity; and gut fullness indices. Results reveal that A. brasiliensis is a typical, generalistic and opportunistic predator that makes use of a wide array of prey types (at least 89 different types), with zooplankton (mainly calanoids), diatoms, terrestrial insects, and plant detritus making up the bulk of the overall diet. The exotic calanoid Temora turbinata ranked as the primary prey. A wide feeding diversity (mean H` = 2.26), low trophic level (mean TROPH = 2.57), and high gut replenishment were persistent across seasons and habitats. Diet composition varied largely and significantly with respect to habitat, season, and body size. A closer assessment showed that habitat and season had a stronger effect on diet than fish size.

Evidence for genetic differentiation of Octopus vulgaris (Mollusca, Cephalopoda) fishery populations from the southern coast of Brazil as revealed by microsatellites

Octopus vulgaris is a cephalopod species in several oceans and commonly caught by artisanal and industrial fisheries. In Brazil, O. vulgaris populations are mainly distributed along the southern coast and have been subjected to intensive fishing during recent years. Despite the importance of this marine resource, no genetic study has been carried out to examine genetic differences among populations along the coast of Brazil. In this study, 343 individuals collected by commercial vessels were genotyped at six microsatellite loci to investigate the genetic differences in O. vulgaris populations along the southern coast of Brazil. Genetic structure and levels of differentiation among sampling sites were estimated via a genotype assignment test and F-statistics. Our results indicate that the O. vulgaris stock consists of four genetic populations with an overall significant analogous F(ST). (phi(CT) = 0.10710, P<0.05) value. The genetic diversity was high with an observed heterozygosity of Ho = 0.987. The negative values of F(IS) found for most of the loci examined suggested a possible bottleneck process. These findings are important for further steps toward more sustainable octopus fisheries, so that this marine resource can be preserved for long-term utilization. (C) 2011 Elsevier B.V. All rights reserved.

Extremely slow feeding in a tropical drilling ectoparasite, Vitularia salebrosa (King and Broderip, 1832) (Gastropoda: Muricidae), on molluscan hosts from Pacific Panama

This study documents one of the slowest feeding behaviors ever recorded for a muricid gastropod in one of the most biotically rigorous regions on the planet. In Pacific Panama, Vitularia salebrosa attacks mollusks by drilling through their shells. The duration of attacks estimated by isotope sclerochronology of oyster shells collected during attacks in progress range from 90 to 230 days, while experimental observation of interactions documented one attack greater than 103 days. The prolonged nature of attacks suggests that V. salebrosa is best characterized as an ectoparasite than as a predator, which is the ancestral condition in the Muricidae. An ectoparasitic lifestyle is also evident in the unusual interaction traces of this species, which include foot scars, feeding tunnels and feeding tubes, specialized soft anatomy, and in the formation of male-female Pairs, which is consistent with protandrous hermaphroditism, as is typical in sedentary gastropods. To delay death of its host, V. salebrosa targets renewable resources when feeding, such as blood and digestive glands. A congener, Vitularia miliaris from the Indo-Pacific, has an identical feeding biology The origin and persistence of extremely slow feeding in the tropics challenges our present understanding of selective pressures influencing the evolution of muricid feeding behaviors and morphological adaptations. Previously, it has been suggested that faster feeding is advantageous because it permits predators to spend a greater proportion of time hiding in enemy-free refugia or to take additional prey, the energetic benefits of which could be translated into increased fecundity or defenses. The benefits of exceptionally slow feeding have received little consideration. In the microhabitat preferred by V. salebrosa (beneath boulders), it is possible that prolonged interactions with hosts decrease vulnerability to enemies by reducing the frequency of risky foraging events between feedings . Ectoparasitic feeding through tunnels by V. salebrosa may also reduce competitive interactions with kleptoparasites (e.g., crabs, snails) that steal food through the gaped valves of dead or dying hosts.

Antibacterial compound from the endophytic fungus Phomopsis longicolla isolated from the tropical red seaweed Bostrychia radicans

Endophytic fungi isolated from the red seaweed Bostrychia radicans were studied to identify their molecularly diverse and biologically active natural chemical products. According to 28S ribosomal DNA-based identification, the strain named C81 was 98% identical to Phomopsis longicolla. This strain was cultivated in solid rice medium and produced three major metabolites identified as 18-deoxycytochalasin H (1), mycophenolic acid (2), and dicerandrol C (3). The chemical structures of these compounds were elucidated by 1D and 2D nuclear magnetic resonance as well as by mass spectrometry. Dicerandrol C had significant antimicrobial activity against Staphylococcus aureus (ATCC 6538) and Staphylococcus saprophyticus (ATCC 15305), with minimum inhibitory concentrations of 1 and 2 mu g ml(-1) (1.33 and 2.66 mu M), respectively. These results show the presence of promising metabolites and indicate that these natural products should be considered in the development of new antibiotics.

Invasion of the dinoflagellate Ceratium furcoides (Levander) Langhans 1925 at tropical reservoir and its relation to environmental variables

Dinoflagellates of the genus Ceratium are chiefly marine but there are rare occurrences in freshwater. In this study we analyze the invasion and progressive establishment of Ceratium furcoides, an exotic species, in the Furnas Reservoir. Samples were taken at 36 points in the reservoir, during the months of March, June, September and December, 2007. Measurements of some physical and chemical variables were simultaneously performed at each site. The occurrence of C. furcoides was registered at 20 sites, with densities varying between 0.57 and 28,564,913.0 ind.m(-3). Blooms of this species were recorded in points which were classified as mesotrophic, coinciding with the places receiving high amounts of untreated domestic sewage. C. furcoides density was correlated with temperature, nutrients (nitrate and nitrite) and water electric conductivity. The highest density was recorded in June when temperature was low. The presence of Ceratium furcoides in the reservoir apparently has not yet affected the reservoir water quality or other plankton communities. However, if it becomes fully established it could perhaps become a problem in the reservoir or even to spread out to other reservoirs in Rio Grande basin.

Evaluation of tropical water sources and mollusks in southern Brazil using microbiological, biochemical, and chemical parameters

Florianopolis, a city located in the Santa Catarina State in southern Brazil, is the national leading producer of bivalve mollusks. The quality of bivalve mollusks is closely related to the sanitary conditions of surrounding waters where they are cultivated. Presently, cultivation areas receive large amounts of effluents derived mainly from treated and non-treated domestic, rural, and urban sewage. This contributes to the contamination of mollusks with trace metals, pesticides, other organic compounds, and human pathogens such as viruses, bacteria, and protozoan. The aim of this study was to perform a thorough diagnosis of the shellfish growing areas in Florianopolis, on the coast of Santa Catarina. The contamination levels of seawater, sediments, and oysters were evaluated for their microbiological, biochemical, and chemical parameters at five sea sites in Florianopolis, namely three regular oyster cultivation areas (Sites 1, 2, and oyster supplier), a polluted site (Site 3), and a heavily polluted site (Site 4). Samples were evaluated at day zero and after 14 days. Seawater and sediment samples were collected just once, at the end of the experiment. Antioxidant defenses, which may occur in contaminated environments in response to the increased production of reactive oxygen species (ROS) by organisms, were analyzed in oysters, as well as organic compounds (in oysters and sediment samples) and microbiological contamination (in oysters and seawater samples). The results showed the presence of the following contaminants: fecal coliforms in seawater samples (four sites), human adenovirus (all sites), human noroviruses GI and GII (two sites), Hepatitis A viruses (one site), JC Polyomavirus in an oyster sample from the oyster supplier, Giardia duodenalis cysts, and Cryptosporidium sp oocysts (one site). Among organochlorine pesticides, only DDT (dichlorodiphenyltrichloroethane) and HCH (hexachlorocyclohexane) were detected in some sediment and oysters samples in very low levels; site 4 had the highest concentrations of total aliphatic hydrocarbons. PAHs, and linear alkylbenzenes (LABs) found either in oysters or in sediment samples. The major concentration of fecal sterol coprostanol was found at site 4, followed by site 3. After 14 days of allocation in the four selected sites, there was a significant difference in the enzymes analyzed at the monitored spots. The detection of different contaminants in oysters, seawater, and sediment samples in the present study shows the impact untreated or inadequately treated effluents have on coastal areas. These results highlight the need for public investment in adequate wastewater treatment and adequate treatment of oysters, ensuring safe areas for shellfish production as well as healthier bivalve mollusks for consumption.

Feeding ecology of the American freshwater goby Ctenogobius shufeldti (Gobiidae, Perciformes) in a sub-tropical estuary

The feeding ecology of the American freshwater goby Ctenogobius shufeldti in a low salinity salt-marsh habitat in the Paranagua Bay estuarine complex (Brazil) was assessed through the gut analysis of 632 individuals. The effects of a set of abiotic factors (type of sediment, salinity, temperature and estuarine reach), season and body size on dietary composition were analysed. Seasonal and size-related changes in feeding strategy, feeding intensity and trophic level were assessed. The effects of gape and body size on prey size use were also analysed. The results showed that C. shufeldti is a typical omnivorous, generalized benthic predator of low trophic levels throughout the seasons and size classes, feeding on 56 dietary items; tanaids, chlorophyte algae, ostracods, gastropods, detritus and benthic diatoms made up the bulk of its diet. The tanaid Kalliapseudes schubarti was the main prey item in both numerical and volumetric terms. The gut fullness was persistently high across the seasons. As expected for a typical generalized, opportunistic omnivorous feeder: (1) seasonal and spatial-temporal variability of abiotic factors had a significant effect on diet structure, (2) season accounted for most of the dietary variation and (3) diet composition and the size of prey consumed did not vary across the size classes.

Reproductive biology of the ghost shrimp Lepidophthalmus bocourti (A. Milne-Edwards, 1870) (Decapoda: Axiidea: Callianassidae): A tropical species with a seasonal reproduction

Based on the hypothesis that reproduction is a continuous process in tropical habitats, we analysed reproductive periodicity and egg production in the callianassid ghost shrimp Lepidophthalmus bocourti, one of the most common species in mangrove systems along the Pacific coast of Central America. During one year (May 2008 to April 2009), individuals of L. bocourti (N = 499) were collected nearshore Gulf of Nicoya, Pacific coast of Costa Rica. Observations were made on presence or absence of incubated embryos, and gonad activity of females was analysed as gonadosomatic index (GSI). Our results revealed that L. bocourti has a marked seasonal breeding period, which contradicts previous reports regarding coastal marine decapods from the tropics. Ovigerous females were found only from June to August, while high GSI values were obtained from March to July. The increase of GSI and appearance of ovigerous females were associated with a concomitant decrease of salinity, but not with temperature. We assume that reproduction of L. bocourti is adapted to local changes of environmental conditions, and that a decrease in salinity during rainy season may serve as a triggering factor for ovarian development. Compared to other ghost shrimps, L. bocourti produced on average more (2002 +/- 1365) and smaller (0.87 +/- 0.109 mm) eggs, which seems to suggest that this species does not have an abbreviated larval development as reported for other species of genus. The deviation from the generalization of constant reproduction in the tropics for shallow water marine invertebrates and its probable cause are adequately discussed.

BIOLOGICAL ASPECTS OF THE SAILFIN DORY Zenopsis conchifer (LOWE, 1852) CAUGHT BY DEEP-SEA TRAWLING FISHERY OFF SOUTHERN BRAZIL

Biological aspects of sailfin dory, Zenopsis conchifer, were studied from 839 individuals obtained from deep-sea commercial bottom trawling off southern Brazil at depths up to 526 m in 2002 and 2003. Samples included fish from 101 mm Lt and 15 g up to 640 mm Lt and 2,9 g. The sex-ratio was 50% at 150 mm Lt and between 300-350 mm Lt, with females outnumbering males in the remaining size classes. Reproductive activity seems to peak between July and August ( austral winter). Size at attainment of 50% maturity (Lt(50)) was 311 mm Lt in females. The mean length and maturity of the specimens increased with depth, suggesting that larger fish concentrate in deeper waters.

Invasion of the dinoflagellate Ceratium furcoides (Levander) Langhans 1925 at tropical reservoir and its relation to environmental variables

Dinoflagellates of the genus Ceratium are chiefly marine but there are rare occurrences in freshwater. In this study we analyze the invasion and progressive establishment of Ceratium furcoides, an exotic species, in the Furnas Reservoir. Samples were taken at 36 points in the reservoir, during the months of March, June, September and December, 2007. Measurements of some physical and chemical variables were simultaneously performed at each site. The occurrence of C. furcoides was registered at 20 sites, with densities varying between 0.57 and 28,564,913.0 ind.m-3. Blooms of this species were recorded in points which were classified as mesotrophic, coinciding with the places receiving high amounts of untreated domestic sewage. C. furcoides density was correlated with temperature, nutrients (nitrate and nitrite) and water electric conductivity. The highest density was recorded in June when temperature was low. The presence of Ceratium furcoides in the reservoir apparently has not yet affected the reservoir water quality or other plankton communities. However, if it becomes fully established it could perhaps become a problem in the reservoir or even to spread out to other reservoirs in Rio Grande basin.

The impact of rising sea temperature on innate immune parameters in the tropical subtidal sea urchin Lytechinus variegatus and the intertidal sea urchin Echinometra lucunter

Ocean temperatures are rising throughout the world, making it necessary to evaluate the impact of these temperature changes on sea urchins, which are well-known bioindicators. This study evaluated the effect of an increase in temperature on the immune response of the subtidal Lytechinus variegatus and the intertidal Echinometra lucunter sea urchins. Both species were exposed to 20 (control), 25 and 30 °C temperatures for 24 h, 2, 7 and 14 days. Counting of coelomocytes and assays on the phagocytic response, adhesion and spreading of coelomocytes were performed. Red and colorless sphere cells were considered biomarkers for heat stress. Moreover, a significant decrease in the phagocytic indices and a decrease in both cell adhesion and cell spreading were observed at 25 and 30 °C for L. variegatus. For E. lucunter, the only alteration observed was for the cell proportions. This report shows how different species of sea urchins respond immunologically to rising temperatures