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).

Characterization of toxic impacts on living marine resources in tidal rivers of the Chesapeake Bay

In 1999, the Chesapeake Bay Program completed a survey of existing data on chemical contaminants and the potential for bioeffects in 38 tidal river systems of Chesapeake Bay. This review led to the identification of 20 areas for which there were insufficient data to adequately characterize the potential for contaminant bioeffects on the Bay’s living resources. The goal of the present study was to estimate the current status of ecological condition in five of these areas and thus help to complete the overall toxics inventory for the Bay. These five systems included the Chester River, Nanticoke River, Pocomoke River, Lower Mobjack Bay (Poquosin and Back Rivers) and the South and Rhode Rivers. This study utilized a Sediment Quality Triad (SQT) approach in combination with additional water-column contaminant analysis to allow for a “weight of evidence” assessment of environmental condition. A total of 60 stations distributed among the five systems, using a probabilistic stratified random design, were sampled during the summer of 2004 to allow for synoptic measures of sediment contamination, sediment toxicity, and benthic condition. Upon completion of all analyses, stations were assigned to one of four categories based on the three legs of the triad. Stations with high sediment quality had no hits on any of the three legs of the triad; those with moderate quality had one hit; those with marginal quality had two hits; and those with poor quality had hits for all three legs of the triad. The Pocomoke River had by far the largest proportion of the total area (97.5%) classified as having high sediment quality, while the Rhode/South system had the highest proportion (11.4%) classified as poor. None of the stations in the Chester River, Nanticoke River, and Lower Mobjack Bay systems were classified as poor. More than 65% of the area of each of the five systems was classified with high to moderate sediment quality. The Rhode/South system had 30.4% of total area classified with marginally to severely poor quality. The results of this study highlight the importance of using multiple indicators and a “weight of evidence” approach to characterize environmental quality and the potential bioeffects of toxic contaminants.

Global transcriptional responses of the toxic cyanobacterium, Microcystis aeruginosa, to nitrogen stress, phosphorus stress, and growth on organic matter

Whole transcriptome shotgun sequencing (RNA-seq) was used to assess the transcriptomic response of the toxic cyanobacterium Microcystis aeruginosa during growth with low levels of dissolved inorganic nitrogen (low N), low levels of dissolved inorganic phosphorus (low P), and in the presence of high levels of high molecular weight dissolved organic matter (HMWDOM). Under low N, one third of the genome was differentially expressed, with significant increases in transcripts observed among genes within the nir operon, urea transport genes (urtBCDE), and amino acid transporters while significant decreases in transcripts were observed in genes related to photosynthesis. There was also a significant decrease in the transcription of the microcystin synthetase gene set under low N and a significant decrease in microcystin content per Microcystis cell demonstrating that N supply influences cellular toxicity. Under low P, 27% of the genome was differentially expressed. The Pho regulon was induced leading to large increases in transcript levels of the alkaline phosphatase phoX, the Pst transport system (pstABC), and the sphX gene, and transcripts of multiple sulfate transporter were also significantly more abundant. While the transcriptional response to growth on HMWDOM was smaller (5–22% of genes differentially expressed), transcripts of multiple genes specifically associated with the transport and degradation of organic compounds were significantly more abundant within HMWDOM treatments and thus may be recruited by Microcystis to utilize these substrates. Collectively, these findings provide a comprehensive understanding of the nutritional physiology of this toxic, bloom-forming cyanobacterium and the role of N in controlling microcystin synthesis.

Porphyrin detection in denatured cnidarian tissue extracts

Porphyrin metabolic disruption from exposure to xenobiotic contaminants such as heavy metals, dioxins, and aromatic hydrocarbons can elicit overproduction of porphyrins. Measurement of porphyrin levels, when used in conjunction with other diagnostic assays, can help elucidate an organism’s physiological condition and provide evidence for exposure to certain toxicants. A sensitive microplate fluorometric assay has been optimized for detecting total porphyrin levels in detergent solubilized protein extracts from symbiotic, dinoflagellate containing cnidarian tissues. The denaturing buffer used in this modified assay contains a number of potentially interfering components (e.g., sodium dodecyl sulfate (SDS), dithiothreitol (DTT), protease inhibitors, and chlorophyll from the symbiotic zooxanthellae), which required examination and validation. Examination of buffer components were validated for use in this porphyrin assay; while the use of a specific spectrofluorometric filter (excitation 400 ± 15 nm; emission 600 ± 20 nm) minimized chlorophyll interference. The detection limit for this assay is 10 fmol of total porphyrin per μg of total soluble protein and linearity is maintained up to 5000 fmol. The ability to measure total porphyrins in a SDS protein extract now allows a single extract to be used in multiple assays. This is an advantage over classical methods, particularly when tissue samples are limiting, as is often the case with coral due to availability and collection permit restrictions.

Compliance Program report of findings: FY 79 pesticides and metals in Fish Program (7305.007)

Because fish bioaccumulate* certain chemicals, levels of chemical contaminants in their edible portion must be closely monitored. In recent years, FDA has conducted several surveys designed to determine the occurrence and levels of selected chemicals or groups of chemicals in fish. Previous fish surveillance programs included the Mercury in Wholesale Fish Survey (FY 71), the FY 73 and 74 Comprehensive Fish Surveys, the Canned Tuna Program (FY 75), the Kepone and Mirex Contamination Program (FY 77), and the FY 77 Mercury in Swordfish Program. In addition, recent Compliance Programs for Pesticides and Metals in Foods and Pesticides, Metals, and Industrial Chemicals in Animal Feed have specified coverage of fish and fish products. Because of previous findings and the sustained high level of fish imported into the United States, a separate compliance program dealing solely with chemical contaminants in fish was initiated by the FDA Bureau of Foods in FY 78. The program includes all domestic and imported fish coverage except that directed by the Bureau of Veterinary Medicine for animal feed components derived from fishery products. The earlier surveys indicated that "bottom feeder" species such as catfish generally had the highest levels of pesticides and polychlorinated biphenyls (PCBs). For this reason, coverage at these species has been emphasized. Similarly, tuna has received special attention because it is the most prevalent fish in the U.S. diet and because of potential problems with mercury. Halibut, swordfish, and snapper also were emphasized in the sampling because of potential problems with mercury levels determined in previous years. The findings in this program were used in detecting emerging problems in fish and directing FDA efforts to deal with them. Care must be exercised in drawing conclusions about trends from the data because this Compliance Program was not statistically designed. Sampling objectives and sources may vary from year to year; thus the results are not directly comparable.

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.

Characterizing Jobos Bay, Puerto Rico: a watershed modeling analysis and monitoring plan

Land-based pollution is commonly identified as a major contributor to the observed deterioration of shallow-water coral reef ecosystem health. Human activity on the coastal landscape often induces nutrient enrichment, hypoxia, harmful algal blooms, toxic contamination and other stressors that have degraded the quality of coastal waters. Coral reef ecosystems throughout Puerto Rico, including Jobos Bay, are under threat from coastal land uses such as urban development, industry and agriculture. The objectives of this report were two-fold: 1. To identify potentially harmful land use activities to the benthic habitats of Jobos Bay, and 2. To describe a monitoring plan for Jobos Bay designed to assess the impacts of conservation practices implemented on the watershed. This characterization is a component of the partnership between the U.S. Department of Agriculture (USDA) and the National Oceanic and Atmospheric Administration (NOAA) established by the Conservation Effects Assessment Project (CEAP) in Jobos Bay. CEAP is a multi-agency effort to quantify the environmental benefits of conservation practices used by private landowners participating in USDA programs. The Jobos Bay watershed, located in southeastern Puerto Rico, was selected as the first tropical CEAP Special Emphasis Watershed (SEW). Both USDA and NOAA use their respective expertise in terrestrial and marine environments to model and monitor Jobos Bay resources. This report documents NOAA activities conducted in the first year of the three-year CEAP effort in Jobos Bay. Chapter 1 provides a brief overview of the project and background information on Jobos Bay and its watershed. Chapter 2 implements NOAA’s Summit to Sea approach to summarize the existing resource conditions on the watershed and in the estuary. Summit to Sea uses a GIS-based procedure that links patterns of land use in coastal watersheds to sediment and pollutant loading predictions at the interface between terrestrial and marine environments. The outcome of Summit to Sea analysis is an inventory of coastal land use and predicted pollution threats, consisting of spatial data and descriptive statistics, which allows for better management of coral reef ecosystems. Chapters 3 and 4 describe the monitoring plan to assess the ecological response to conservation practices established by USDA on the watershed. Jobos Bay is the second largest estuary in Puerto Rico, but has more than three times the shoreline of any other estuarine area on the island. It is a natural harbor protected from offshore wind and waves by a series of mangrove islands and the Punta Pozuelo peninsula. The Jobos Bay marine ecosystem includes 48 km² of mangrove, seagrass, coral reef and other habitat types that span both intertidal and subtidal areas. Mapping of Jobos Bay revealed 10 different benthic habitats of varying prevalence, and a large area of unknown bottom type covering 38% of the entire bay. Of the known benthic habitats, submerged aquatic vegetation, primarily seagrass, is the most common bottom type, covering slightly less than 30% of the bay. Mangroves are the dominant shoreline feature, while coral reefs comprise only 4% of the total benthic habitat. However, coral reefs are some of the most productive habitats found in Jobos Bay, and provide important habitat and nursery grounds for fish and invertebrates of commercial and recreational value.

Toxic effect of chalkones and antitoxic role of ascorbic acid

One of the chalkones synthesised in the author's laboratory was selected to determine its toxicity to fish, Lepidocephalicthys thermalis at different concentrations and time periods. Ascorbic acid contents were determined and it was found to be antitoxic.

Toxicity of heavy metals to brine shrimp Artemia

Bioassay were carried out on 48h cultured nauplii of brine shrimp Artemia by exposing them to seven trace elements viz. copper (Cu), zinc (Zn), cadmium (Cd), nickel (Ni), lead (Pb), iron (Fe) and manganese (Mn). Synergistic effects of all these elements and additive effects of Cu and Zn, Cd and Pb, and Ni and Fe were also investigated. Comparatively, the degree of toxicity for compound bioassays was higher than individual simple tests. The values were averaged and the expected median lethal concentration [LC sub(50)] of tested heavy metals was obtained by probit analysis on the basis of cumulative numbers of dead organisms after 24 and 48h. The order of toxicity of the metals to Artemia was Pb>Cd>Cu>Ni>Zn>Fe>Mn. Potency ratios of the seven metals were also calculated. The 24 and 48h variations obtained in LC sub(50) values were significantly different and relative implications of these are discussed.

Toxic effects of dietary seaweeds Sargassum bovianum, Caulera faridii and Gracilaria corticata on blood parameters of albino rats

The present communication deals with the feeding trials of brown (Sargassum bovianum), green (Caulerpa faridii) and red (Gracilaria corticola) seaweeds in albino rats for a period of thirty days in order to investigate their digestibility and acceptability as supplementary food for animals. The parameters used were: changes in blood hemoglobin, ESR, MCHC, PCV and plasma vitamin levels. The result revealed that all the three species of seaweeds had acceptability up to 5% level, as no ill effect was noted during the experiment. But at 10% and 20% levels, marked changes were observed in blood parameters with diarrhea, vomiting and convulsions indicating possibilities of either tissue and muscular dystrophy, gastrointestinal tract necrosis or functional disorder of central nervous system. A heavy mortality was noted due to excessive water loss through diarrhea and vomiting. However, no mortality was observed after 22nd day at both 10% and 20% levels with subsided clinical signs. The results suggest that these three seaweed species could be used safely as a supplementary food, in native form, in animals at low concentrations.

Toxic effects of dimecron to fingerlings of Labeo rohita (Ham.) during acute and chronic exposures

The toxic effects of dimecron on growth, body composition and oxygen consumption of fingerlings of Labeo rohita were studied. Dimecron concentrations of 4 and 8 mg/l were used. Both acute (3-h) and chronic (15- 42 d) exposure schedules were followed. Compared with the control fish, both 4 and 8 mg/l dimecron treatment significantly suppressed weight gained in fish by 9.71% and 30% respectively during a 42 day exposure period. However, the length of fish was suppressed by 11.46% significantly only in fish group exposed to 8 mg/l dimecron. Similarly, the protein content was also significantly reduced in the above group of fish. The oxygen consumption of fish was elevated considerably, but not significantly in both group of treated fish (8.5% and 26.07%) during acute exposure. However, after 15 days of exposure the rate decreased by 18.98% significantly only in fish exposed to 8 mg/l dimecron. The threshold level of DO at low oxygen environment found to be slightly higher in fish at 8 mg/l dimecron. The survival time at the above oxygen condition was reduced during acute exposure (3-h) and that was extended during chronic (15-d) exposure.

Circulation system and areas of potentially successful tuna fishing with surface methods off Mozambique

Charts with 3-months running means of thermal data collected during 1977-1982 are used to describe the seasonal variability of the circulation pattern off Mozambique, and to identify the areas of tuna vulnerability to surface gears. The main conclusions reached by Sætre and Jorge da Silva (1984) have received further support. Areas of potentially successful exploitation of tunas by surface methods have been identified during the whole year, with emphasis for November-April.

Acute toxicity of copper, zinc and mercury on intertidal gastropods of Mumbai coast

The acute toxicity test conducted by static bioassay techniques have revealed that among selected heavy metals, copper is more toxic than zinc and mercury to Planaxis sulcatus and Trochus radiatus. The natural availability of heavy metals in the surrounding environment of these organisms is found to be deciding factor for their toxicity. Natural habitat of the animal also contributes to the sensitivity of a particular animal to the heavy metals tested. In addition the tendency of the animal to overcome the adverse conditions in their surrounding also plays a significant role in toxicity of pollutants.

Toxic effects of dimecron to Labeo rohita Ham. fingerlings during acute and chronic exposures: histopathological changes

The exposure to the highest dimecron cone. (8 mg/1) resulted in severe histopathological changes in different tissues of Labeo rohita fingerling. Cell necrosis, cytoplasmic vacuolation and pycnotic nuclei were major abnormalities observed in liver tissue. The degeneration of glomeruli and proximal tubules, cytoplasmic vacuolation and focal haemorrhagic area were noted in case of kidney tissues. Major changes observed in intestinal tissues were degeneration of villi, disintegrity of mucosal layers, necrosis of epithelial cells etc. However, hypertrophy of cells and granulation of cytoplasm were major histopathological changes observed in fish at lower dimecron cones. (4 mg/1).