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).

Flux and sources of nutrients in the Mississippi-Atchafalaya River Basin: Topic 3 Report for the Integrated Assessment on Hypoxia in the Gulf of Mexico.

Ths report addresses the following two questions: 1) What are the loads (flux) of nutrients transported from the Mississippi-Atchafalaya River Basin to the Gulf of Mexico, and where do they come from within the basin? 2) What is the relative importance of specific human activities, such as agriculture, point-source discharges, and atmospheric deposition in contributing to these loads? These questions were addressed by first estimating the flux of nutrients from the Mississippi-Atchafalaya River Basin and about 50 interior basins in the Mississippi River system using measured historical streamflow and water quality data. Annual nutrient inputs and outputs to each basin were estimated using data from the National Agricultural Statistics Service, National Atmospheric Deposition Program, and point-source data provided by the USEPA. Next, a nitrogen mass balance was developed using agricultural statistics, estimates of nutrient cycling in agricultural systems, and a geographic information system. Finally, multiple regression models were developed to estimate the relative contributions of the major input sources to the flux of nitrogen and phosphorus to the Gulf of Mexico.

Change in Pacific Northwest coastal ecosystems. Proceedings of the Pacific Northwest Coastal Ecosystems Regional Study Workshop, August 73-14, 1996, Troutdale, Oregon

Over the past one hundred and fifty years, the landscape and ecosystems of the Pacific Northwest coastal region, already subject to many variable natural forces, have been profoundly affected by human activities. In virtually every coastal watershed from the Strait of Juan de Fuca to Cape Mendocino, settlement, exploitation and development of resou?-ces have altered natural ecosystems. Vast, complex forests that once covered the region have been largely replaced by tree plantations or converted to non-forest conditions. Narrow coastal valleys, once filled with wetlands and braided streams that tempered storm runoff and provided salmon habitat, were drained, filled, or have otherwise been altered to create land for agriculture and other uses. Tideflats and saltmarshes in both large and small estuaries were filled for industrial, commercial, and other urban uses. Many estuaries, including that of the Columbia River, have been channeled, deepened, and jettied to provide for safe, reliable navigation. The prodigious rainfall in the region, once buffered by dense vegetation and complex river and stream habitat, now surges down sirfiplified stream channels laden with increased burdens of sediment and debris. Although these and many other changes have occurred incrementally over time and in widely separated areas, their sum can now be seen to have significantly affected the natural productivity of the region and, as a consequence, changed the economic structure of its human communities. This activity has taken place in a region already shaped by many interacting and dynamic natural forces. Large-scale ocean circulation patterns, which vary over long time periods, determine the strength and location of currents along the coast, and thus affect conditions in the nearshore ocean and estuaries throughout the region. Periodic seasonal differences in the weather and ocean act on shorter time scales; winters are typically wet with storms from the southwest while summers tend to be dry with winds from the northwest. Some phenomena are episodic, such as El Nifio events, which alter weather, marine habitats, and the distribution and survival of marine organisms. Other oceanic and atmospheric changes operate more slowly; over time scales of decades, centuries, and longer. Episodic geologic events also punctuate the region, such as volcanic eruptions that discharge widespread blankets of ash, frequent minor earthquakes, and major subduction zone earthquakes each 300 to 500 years that release accumulated tectonic strain, dropping stretches of ocean shoreline, inundating estuaries and coastal valleys, and triggering landslides that reshape stream profiles. While these many natural processes have altered, sometimes dramatically, the Pacific Northwest coastal region, these same processes have formed productive marine and coastal ecosystems, and many of the species in these systems have adapted to the variable environmental conditions of the region to ensure their long-term survival.

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.

Population structure of island-associated dolphins: evidence from photo-identification of common bottlenose dolphins (Tursiops truncatus) in the main Hawaiian Islands

Management agencies often use geopolitical boundaries as proxies for biological boundaries. In Hawaiian waters a single stock is recognized of common bottlenose dolphins, Tursiops truncatus, a species that is found both in open water and near-shore among the main Hawaiian Islands. To assess population structure, we photo-identified 336 distinctive individuals from the main Hawaiian Islands, from 2000 to 2006. Their generally shallow-water distribution, and numerous within-year and between-year resightings within island areas suggest that individuals are resident to the islands, rather than part of an offshore population moving through the area. Comparisons of identifications obtained from Kaua‘i/Ni‘ihau, O‘ahu, the “4-island area,” and the island of Hawai‘i showed no evidence of movements among these island groups, although movements from Kaua‘i to Ni‘ihau and among the “4-islands” were documented. A Bayesian analysis examining the probability of missing movements among island groups, given our sample sizes for different areas, indicates that interisland movement rates are less than 1% per year with 95% probability. Our results suggest the existence of multiple demographically independent populations of island-associated common bottlenose dolphins around the main Hawaiian islands.

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.

Azaspiracid shellfish poisoning: a review on the chemistry, ecology, and toxicology with an emphasis on human health impacts

Azaspiracids (AZA) are polyether marine toxins that accumulate in various shellfish species and have been associated with severe gastrointestinal human intoxications since 1995. This toxin class has since been reported from several countries, including Morocco and much of western Europe. A regulatory limit of 160 μg AZA/kg whole shellfish flesh was established by the EU in order to protect human health; however, in some cases, AZA concentrations far exceed the action level. Herein we discuss recent advances on the chemistry of various AZA analogs, review the ecology of AZAs, including the putative progenitor algal species, collectively interpret the in vitro and in vivo data on the toxicology of AZAs relating to human health issues, and outline the European legislature associated with AZAs.

Domoic acid contamination within eight representative species from the benthic food web of Monterey Bay, California, USA

Benthic food webs often derive a significant fraction of their nutrient inputs from phytoplankton in the overlying waters. If the phytoplankton include harmful algal species like Pseudo-nitzschia australis, a diatom capable of producing the neurotoxin domoic acid (DA), the benthic food web can become a depository for phycotoxins. We tested the general hypothesis that DA contaminates benthic organisms during local blooms of P. australis, a widespread toxin producer along the US west coast. To test for trophic transfer and uptake of DA into the benthic food web, we sampled 8 benthic species comprising 4 feeding groups: filter feeders (Emerita analoga and Urechis caupo); a predator (Citharichthys sordidus); scavengers (Nassarius fossatus and Pagurus samuelis) and deposit feeders (Neotrypaea californiensis, Dendraster excentricus and Olivella biplicata). Sampling occurred before, during and after blooms of P. australis in Monterey Bay, CA, USA during 2000 and 2001. DA was detected in all 8 species, with contamination persisting over variable time scales. Maximum DA levels in N. fossatus (674 ppm), E. analoga (278 ppm), C. sordidus (515 ppm), N. californiensis (145 ppm), P. samuelis (56 ppm), D. excentricus (15 ppm) and O. biplicata (3 ppm) coincided with P. australis blooms, while DA levels in U. caupo remained above 200 ppm (max. = 751 ppm) throughout the study period. DA in 6 species exceeded levels thought to be safe for higher level consumers (i.e. ≥20 ppm) and thus is likely to have deleterious effects on marine birds, sea lions and the endangered California sea otter, known to prey upon these benthic species.

Field applications of the second-generation Environmental Sample Processor (ESP) for remote detection of harmful algae: 2006-2007

We assess the application of the second-generation Environmental Sample Processor (ESP) for the detection of harmful algal bloom (HAB) species in field and laboratory settings using two molecular probe techniques: a sandwich hybridization assay (SHA) and fluorescent in situ hybridization (FISH). During spring 2006, the first time this new instrument was deployed, the ESP successfully automated application of DNA probe arrays for various HAB species and other planktonic taxa, but non-specific background binding on the SHA probe array support made results interpretation problematic. Following 2006, the DNA array support membrane that we were using was replaced with a different membrane, and the SHA chemistry was adjusted. The sensitivity and dynamic range of these modifications were assessed using 96-well plate and ESP array SHA formats for several HAB species found commonly in Monterey Bay over a range of concentrations; responses were significantly correlated (p < 0.01). Modified arrays were deployed in 2007. Compared to 2006, probe arrays showed improved signal:noise, and remote detection of various HAB species was demonstrated. We confirmed that the ESP and affiliated assays can detect HAB populations at levels below those posing human health concerns, and results can be related to prevailing environmental conditions in near real-time.

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.

Abundance of cetatceans in the southern U.S. North Atlantic Ocean during summer 1998

The U.S. Marine Mammal Protection Act requires that the abundance of marine mammals in U.S. waters be assessed. Because this requirement had not been met for a large portion of the North Atlantic Ocean (U.S. waters south of Maryland), a ship-based, line-transect survey was conducted with a 68 m research ship between Maryland (38.00°N) and central Florida (28.00°N) from the 10-m isobath to the boundary of the U.S. Exclusive Economic Zone. The study area (573,000 km2) was surveyed between 8 July and 17 August 1998. Minimum abundance estimates were based on 4163 km of effort and 217 sightings of at least 13 cetacean species and other taxonomic categories. The most commonly sighted species (number of groups) were bottlenose dolphins, Tursiops truncatus (38); sperm whales, Physeter macrocephalus (29); Atlantic spotted dolphins, Stenella frontalis (28); and Risso’s dolphins, Grampus griseus (22). The most abundant species (abundance; coeffi cient of variation) were Atlantic spotted dolphins (14,438; 0.63); bottlenose dolphins (13,085; 0.40); pantropical spotted dolphins, S. attenuate (12,747; 0.56); striped dolphins, S. coeruleoalba (10,225; 0.91); and Risso’s dolphins (9533; 0.50). The abundance estimate for the Clymene dolphin, S. clymene (6086; 0.93), is the first for the U.S. Atlantic Ocean. Sperm whales were the most abundant large whale (1181; 0.51). Abundances for other species or taxonomic categories ranged from 20 to 5109. There were an estimated 77,139 (0.23) cetaceans in the study area. Bottlenose dolphins and Atlantic spotted dolphins were encountered primarily in continental shelf (<200 m) and continental slope waters (200−2000 m). All other species were generally sighted in oceanic waters (>200 m). The distribution of some species varied north to south. Striped dolphins, Clymene dolphins, and sperm whales were sighted primarily in the northern part of the study area; whereas pantropical spotted dolphins were sighted primarily in the southern portion.

Age and growth of the blue shark (Prionace glauca) in the North Atlantic Ocean

Age and growth estimates for the blue shark (Prionace glauca) were derived from 411 vertebral centra and 43 tag-recaptured blue sharks collected in the North Atlantic, ranging in length from 49 to 312 cm fork length (FL). The vertebrae of two oxytetracycline-injected recaptured blue sharks support an annual spring deposition of growth bands in the vertebrae in sharks up to 192 cm FL. Males and females were aged to 16 and 15 years, respectively, and full maturity is attained by 5 years of age in both sexes. Both sexes grew similarly to age seven, when growth rates decreased in males and remained constant in females. Growth rates from tag-recaptured individuals agreed with those derived from vertebral annuli for smaller sharks but appeared overestimated for larger sharks. Von Bertalanffy growth parameters derived from vertebral length-at-age data are L∞ = 282 cm FL, K = 0.18, and t0 = –1.35 for males, and L∞ = 310 cm FL, K = 0.13, and t0 = −1.77 for females. The species grows faster and has a shorter life span than previously reported for these waters.