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Aquatic macroaggregates (flocs ≥ 0.5 mm) provide an important mechanism for vertical flux of nutrients and organic matter in aquatic ecosystems, yet their role in the transport and fate of zoonotic pathogens is largely unknown. Terrestrial pathogens that enter coastal waters through contaminated freshwater runoff may be especially prone to flocculation due to fluid dynamics and electrochemical changes that occur where fresh and marine waters mix. In this study, laboratory experiments were conducted to evaluate whether zoonotic pathogens (Cryptosporidium, Giardia, Salmonella) and a virus surrogate (PP7) are associated with aquatic macroaggregates and whether pathogen aggregation is enhanced in saline waters. Targeted microorganisms showed increased association with macroaggregates in estuarine and marine waters, as compared with an ultrapure water control and natural freshwater. Enrichment factor estimations demonstrated that pathogens are 2-4 orders of magnitude more concentrated in aggregates than in the estuarine and marine water surrounding the aggregates. Pathogen incorporation into aquatic macroaggregates may influence their transmission to susceptible hosts through settling and subsequent accumulation in zones where aggregation is greatest, as well as via enhanced uptake by invertebrates that serve as prey for marine animals or as seafood for humans.

The neurotoxin domoic acid (DA) produced by the diatoms Pseudo-nitzschia has been responsible for deaths of marine mammals and birds in Monterey Bay, California, USA. In this study we examined links between DA-related strandings of the seasonally migratory California sea lion Zalophus californianus and regional occurrences of DA-producing diatoms using a decade-long time series. Results suggest a more complex pattern of stranding than anticipated, one not related simply to regional abundance of the toxin producers. Stranding patterns of sea lions exhibiting signs of acute DA toxicosis may be best explained by multiple causative factors including timing of toxic blooms with respect to the sea lion breeding cycle, adequacy of sea lion prey during the breeding season, as well as the geographic range of this pinniped outside of the breeding season. Three DA-related stranding events occurred in Monterey Bay in 1998, 2000 and 2007, when toxic DA blooms were pre- sent in the Bay and the California coast was experiencing El Nino/Southern Oscillation conditions. Foraging centers near the breeding site likely provided inadequate food for individuals, leading to northerly movement of these highly mobile predators to other geographic sites resulting in exposure to toxic DA-producing blooms. High toxic Pseudo-nitzschia and DA concentrations did not, however, result in DA-related stranding events in 2002, 2003 and 2004, when weaker than normal upwelling events were present in the Bay. Relative productivity of central versus southern California with respect to the breeding season thus appears to strongly influence the frequency of DA-related strand- ings in Monterey Bay, California.

Domoic acid is a glutaminergic neurotoxin produced by marine algae such as Pseudo-nitzschia australis. California sea lions (Zalophus californianus) ingest the toxin when foraging on planktivorous fish. Adult females comprise 60% of stranded animals admitted for rehabilitation due to acute domoic acid toxicosis and commonly suffer from reproductive failure, including abortions and premature live births. Domoic acid has been shown to cross the placenta exposing the fetus to the toxin. To determine whether domoic acid was playing a role in reproductive failure in sea lion rookeries, 67 aborted and live-born premature pups were sampled on San Miguel Island in 2005 and 2006 to investigate the causes for reproductive failure. Analyses included domoic acid, contaminant and infectious disease testing, and histologic examination. Pseudo-nitzschia spp. were present both in the environment and in sea lion feces, and domoic acid was detected in the sea lion feces and in 17% of pup samples tested. Histopathologic findings included systemic and localized inflammation and bacterial infections of amniotic origin, placental abruption, and brain edema. The primary lesion in five animals with measurable domoic acid concentrations was brain edema, a common finding and, in some cases, the only lesion observed in aborted premature pups born to domoic acid-intoxicated females in rehabilitation. Blubber organochlorine concentrations were lower than those measured previously in premature sea lion pups collected in the 1970s. While the etiology of abortion and premature parturition was varied in this study, these results suggest that domoic acid contributes to reproductive failure on California sea lion rookeries.

In California, the toxic algal species of primary concern are the dinoflagellate Alexandrium catenella and members of the pennate diatom genus Pseudo-nitzschia, both producers of potent neurotoxins that are capable of sickening and killing marine life and humans. During the summer of 2004 in Monterey Bay, we observed a change in the taxonomic structure of the phytoplankton community—the typically diatom-dominated community shifted to a red tide, dinoflagellate-dominated community. Here we use a 6-year time series (2000–2006) to show how the abundance of the dominant harmful algal bloom (HAB) species in the Bay up to that point, Pseudo-nitzschia, significantly declined during the dinoflagellate-dominated interval, while two genera of toxic dinoflagellates, Alexandrium and Dinophysis, became the predominant toxin producers. This change represents a shift from a genus of toxin producers that typically dominates the community during a toxic bloom, to HAB taxa that are generally only minor components of the community in a toxic event. This change in the local HAB species was also reflected in the toxins present in higher trophic levels. Despite the small contribution of A. catenella to the overall phytoplankton community, the increase in the presence of this species in Monterey Bay was associated with an increase in the presence of paralytic shellfish poisoning (PSP) toxins in sentinel shellfish and clupeoid fish. This report provides the first evidence that PSP toxins are present in California's pelagic food web, as PSP toxins were detected in both northern anchovies (Engraulis mordax) and Pacific sardines (Sardinops sagax). Another interesting observation from our data is the co-occurrence of DA and PSP toxins in both planktivorous fish and sentinel shellfish. We also provide evidence, based on the statewide biotoxin monitoring program, that this increase in the frequency and abundance of PSP events related to A. catenella occurred not just in Monterey Bay, but also in other coastal regions of California. Our results demonstrate that changes in the taxonomic structure of the phytoplankton community influences the nature of the algal toxins that move through local food webs and also emphasizes the importance of monitoring for the full suite of toxic algae, rather than just one genus or species.

Benthic food webs often derive a significant fraction of their nutrient inputs from phyto- plankton 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 contami- nates 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. Max- imum DA levels in N. fossatus (674 ppm), E. analoga (278 ppm), C. sordidus (515 ppm), N. californien- sis (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) through- out 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 endan- gered California sea otter, known to prey upon these benthic species.

Yessotoxin (YTX) was detected in an algal sample and two mussel samples (0.07–0.10 μg g −1 ) collected from Scripps Pier in La Jolla, California during a bloom of Lingulodinium polyedrum . Mussel samples collected from Monterey Bay, California also contained measurable YTX (levels up to 0.06 μg g −1 ) in samples obtained during a 6-month (weekly) sampling period. Gonyaulax spinifera and L. polyedrum were identified in background concentrations in Monterey Bay during the time of contamination. An algal sample from Washington coastal waters collected during non-bloom conditions also contained YTX, possibly originating from Protoceratium reticulatum . Three strains of L. polyedrum (CCMP1931, CCMP1936, 104A) isolated from southern California coastal waters and one strain of G. spinifera (CCMP409) isolated from Maine were tested for YTX production using two methods, competitive ELISA and liquid chromatography–mass spectrometry (LC–MS). The ELISA method detected YTX in the particulate phase in two of three L. polyedrum strains. The LC–MS method did not detect YTX in the particulate or dissolved phase of any of the strains. To our knowledge, this is the first study to test and confirm YTX in environmental samples from California and Washington coastal waters. It is highly likely that L. polyedrum was responsible for the YTX contamination in the southern California samples. Future research needs to conclusively determine the biological origin(s) of YTX contamination in central California and Washington coastal waters.

Over the past decade diatom blooms of domoic acid (DA)-producing Pseudo-nitzschia spp. have been responsible for numerous marine mammal and bird mortalities in Monterey Bay, CA. One possible toxin vector is the market squid, Loligo opalescens , a small pelagic mollusk that plays an important role in the near-shore food web of the California Current ecosystem as a favored vertebrate prey species. This study examined the trophic link between toxic Pseudo-nitzschia and L. opalescens using toxin and stomach content analyses of animals collected from Monterey Bay, CA in 2000. Receptor binding assay data (confirmed by tandem mass spectrometry), demonstrated the presence of DA in squid during a toxic Pseudo-nitzschia event, with P. australis frustules observed in stomach samples. Though DA levels were low ( −1 tissue) in L. opalescens during the study period, it is now clear that this potent neurotoxin can occur in squid and is likely delivered through its krill prey species, which are known to retain DA after feeding on toxic Pseudo-nitzschia . Our findings suggest that further study of the relationship between Pseudo-nitzschia blooms and DA contamination of squid is warranted to better evaluate the potential health risk to humans and wildlife associated with this major commercial seafood species and important prey item.

Within Monterey Bay, California, USA, the food web transfer of domoic acid (DA), a neurotoxin produced by diatoms of the genus Pseudo-nitzschia, has led to major mortality events of marine mammals and birds. Less visible, and less well known, is whether invertebrates and fish associated with the benthos are also affected by blooms of DA-producing Pseudo-nitzschia spp. This study examines the presence of DA in benthic flatfish offshore of Davenport, California, (37°0′36″N, 122°13′12″W) and within Monterey Bay, California (36°45′0″N, 122°1′48″W), including species that feed primarily in the sediment (benthic-feeding) and species that feed primarily in the water column (benthopelagic-feeding). Flatfish caught between 10 December 2002 and 17 November 2003 at depths of 30–180 m had concentrations of DA in the viscera ranging from 3 to 26 μg DA g−1 of viscera. Although the DA values reported are relatively low, benthic-feeding flatfish were frequently contaminated with DA, especially as compared with the frequency of contamination of flatfish species that feed in the water column. Furthermore, on days in which both benthic-feeding and benthopelagic-feeding flatfish were collected, the former had significantly higher concentrations of DA in the viscera. Curlfin turbot, Pleuronicthys decurrens, the flatfish with both the highest level and frequency of DA contamination, are reported to feed exclusively on polychaetes, suggesting that these invertebrates may be an important vector of the toxin in benthic communities and may pose a risk to other benthic-feeding organisms.

Blooms of domoic acid (DA) synthesizing diatoms (Pseudo-nitzschia spp.) have been associated with the death and injury of hundreds of marine shorebirds and mammals, exposed humans to potentially serious health risks, and threatened to significantly impact coastal fisheries and commerce dependent on marine resources. While indicator organisms are widely utilized to monitor for marine biotoxins like paralytic shellfish poisoning toxins, a reliable intertidal indicator species to monitor DA remains to be identified. Here we evaluate and confirm the utility of the common sand crab (Emerita analoga) as an indicator for DA in comparison with sea mussels (Mytilus californianus). Mussels and sand crabs, collected from natural populations in Santa Cruz, California (April 1999-February 2000), were tested for DA using the HPLC-UV method. Toxin loads in sand crabs ranged from below detectable limits to 13.4 micro g DA g(-1) and coincided with the abundance of DA producing Pseudo-nitzschia species nearshore. Toxin levels in mussels collected during the study period were below HPLC-UV detectable limits. The rise and fall of DA in sand crabs in synchrony with Pseudo-nitzschia abundance, combined with this common intertidal species' accessibility and ease of DA extraction, clearly indicate the utility of sand crabs as a reliable, cost-effective monitoring tool for DA in the nearshore coastal environment.

In at least two mass mortality events in Monterey Bay, California, planktivorous fish were implicated as vectors of the neurotoxin domoic acid (DA) from diatoms to sea birds and marine mammals. Whereas the transfer of DA from planktivorous fish to piscivorous predators has been well established, the relationship between toxin levels in plankton-feeding fish and the regional abundance of DA-producing diatoms has not been documented. Here we present data from an extensive field study in which cell densities of toxic Pseudo-nitzschia species and DA levels in anchovies and sardines were measured from samples collected weekly throughout Monterey Bay from 8 October 1999 to 18 October 2000. Four distinct blooms were documented with cell densities ranging from 3.2×103 to 5.0×105 cells l–1. DA was detected in fish viscera samples whenever toxic diatom densities reached ≥103 cells l–1 in surface waters, suggesting that anchovies and sardines regularly consume toxic diatoms when present. Fish contained DA levels above the regulatory limit (20 µg DA g–1 whole fish) only when toxic cell densities exceeded 104 cells l–1. DA was only detected in fish when toxic diatom species were also present in the water, suggesting that the toxin is quickly depurated and that fish are only dangerous vectors during the bloom period. Anchovies appear to be more potent vectors than sardines as they consistently contained more DA than sardines collected simultaneously. Maximum DA levels detected in fish were 1,815 µg DA g–1 in anchovy and 728 µg DA g–1 in sardine viscera samples. In fish with high viscera levels of DA, corresponding body tissues contained 0.2–2.2 µg DA g–1 (0.2±0.1% of the viscera level), suggesting that DA is not accumulated in edible body tissues to levels that threaten human consumers. Results from this study suggest that anchovies may be a valuable indicator species for assessing the risk of DA intoxication to piscivorous sea birds and mammals during toxic Pseudo-nitzschia blooms in Monterey Bay.

Over the past decade, blooms of the domoic acid (DA) producing diatom Pseudo- nitzschia have been responsible for numerous deaths of marine mammals and birds in Monterey Bay, California. Euphausiids (krill) are important members of the local zooplankton grazer community and comprise the primary diet of squid, baleen whales, and many seabirds. Krill are thus a key potential vector for the transfer of DA to higher trophic level organisms in Monterey Bay. A better understand- ing of the quantitative trophic interactions and body burden of DA in krill is required to predict whether they can act as an effective vector for this neurotoxin. Here we report results of toxin analy- ses and gut content examinations of krill Euphausia pacifica collected from Monterey Bay in 2000. Corresponding counts of toxic Pseudo-nitzschia species in the water and their cellular DA concen- trations were also obtained at the collection sites. Toxin analysis by receptor binding assay demon- strated that DA in krill tissue varied between 0.1 to 44 µg DA equiv. g -1 tissue (confirmed by tandem mass spectrometry), with levels corresponding to the abundance of toxic Pseudo-nitzschia species present in the water. The occurrence of Pseudo-nitzschia australis frustules in the digestive tract of E. pacifica verified that a toxic species of this diatom was an important part of their diet and thus impli- cated this phytoplankter as the source of DA. These findings provide compelling evidence for the role of krill as a potential transfer agent of the phycotoxin DA to higher trophic levels in marine food web.

The planktivorous northern anchovy is a prominent vector of the phycotoxin domoic acid (DA) to organisms at higher trophic levels, including fish-eating seabirds and mammals. Although there are abundant data reporting DA-induced excitotoxic symptoms in higher vertebrates, to date there has been no reported evidence of neurotoxic effects in lower vertebrate vectors such as fish. To explain this apparent lack of toxicity, it has been suggested that DA may not reach the brain in anchovies and/or that fish are not as sensitive neurologically to DA. In the present study, intracoelomic (IC) injection of DA, at doses ranging from 1 to 14 μg DA g−1 total fish weight, resulted in severe neurotoxic symptoms such as spinning, disorientation, inability to school, and mortality, indicating that anchovies are neurologically susceptible and that DA crosses the blood–brain barrier in fish. An ED50 of 3.2 μg DA g−1 total body weight was determined via IC injection of DA in 83 anchovies. Comparable intraperitoneal injection studies with mice, rats, and monkeys report similar DA-induced neurotoxic symptoms at doses near 3.2 μg DA g−1, suggesting a similar neurologic sensitivity and mechanism of toxicity between anchovies and mammals. DA tissue distribution measurements from freshly collected field-exposed anchovies and orally gavaged anchovies indicate that DA uptake from the gastrointestinal tract does occur. Levels as high as 1,175 μg DA g−1 were measured in anchovy viscera, while muscle and brain tissue DA levels were 3 orders of magnitude lower, indicating low but measurable DA uptake. Further evidence is needed to confirm that uptake is sufficient during field events to induce symptoms in anchovies. Our work provides the first reported evidence of neurotoxic symptoms in fish and suggests that anchovies may be affected by DA during toxic diatom blooms. If sufficient uptake occurs, DA-induced neurotoxic symptoms and mortality may make fish easier prey targets, thereby selecting for the highest toxin levels transferred, as well as providing a possible pathway for the transfer of DA to benthic communities.

The occurrence of an unusual mortality event involving California sea lions (Zalophus californianus) along the central California coast in May 1998 was recently reported. The potent neurotoxin domoic acid (DA), produced naturally by the diatom Pseudo-nitzschia australis and transmitted to the sea lions via planktivorous northern anchovies (Engraulis mordax), was identified as the probable causative agent. Details of DA analyses for anchovy tissues and sea lion feces are described. Domoic acid levels were estimated in anchovy samples by HPLC-UV, and in sea lion feces using the same method as well as a microplate receptor binding assay, with absolute confirmation by tandem mass spectrometry. The highest DA concentrations in anchovies occurred in the viscera (223 +/- 5 microg DA g(-1)), exceeding values in the body tissues by seven-fold and suggesting minimal bioaccumulation of DA in anchovy tissue. HPLC values for DA in sea lion fecal material (ranging from 152 to 136.5 microg DA g(-1)) required correction for interference from an unidentified compound. Inter-laboratory comparisons of HPLC data showed close quantitative agreement. Fecal DA activity determined using the receptor binding assay corresponded with HPLC values to within a factor of two. Finally, our detection of P. australis frustules, via scanning electron microscopy, in both anchovy viscera and fecal material from sea lions exhibiting seizures provides corroborating evidence that this toxic algal species was involved in this unusual sea lion mortality event.

Bacterioplankton abundance and metabolic characteristics were observed in northern San Francisco Bay, California, during spring and summer 1996 at three sites: Central Bay, Suisun Bay, and the Sacramento River. These sites spanned a salinity gradient from marine to freshwater, and sampling occurred during a period of seasonally declining river flow. The microbial measures included radio-labeled amino acid uptake ( L-leucine, L-proline, Lserine), ectoenzyme activity (aminopeptidase and b-D-glucosidase), and bacterial abundance using 1-mm filters to separate free from particle-associated bacteria. A seasonal decline in all bacterial metabolic measures was observed at all stations, suggesting that a system-wide variable may be important in controlling bacterial activity. One such variable is freshwater flow into the Bay (as a proxy for organic matter flux), which positively covaried with all metabolic measures. A sharp decline in particle-associated bacteria was also observed in Suisun Bay and the Sacramento River between July and August. This decline may have been due to combined effects of declining nutritive value of the aging particles and increasing grazing pressure by benthic filter feeders. Aminopeptidase activity was positively related with increasing salinity, and b-D-glucosidase was negatively correlated with increasing salinity, indicating a gradient in the relative quality of organic matter from carbohydrate-rich riverine to proteinrich oceanic material. Overall, Suisun Bay had the highest mean proportion of particle-associated bacteria (49%), followed by Sacramento River (36%) and Central Bay (11%). Particles were the sites of enhanced ectoenzyme activity but not amino acid incorporation. Bacteria may be actively dissolving the particulate organic matter, but their growth rates on particles are not significantly enhanced.

During the autumn of 1991, numerous seabird fatalities in Monterey Bay, California, led to the discovery of a new domoic acid-producing diatom, Pseudonitzschia australis. Since this initial event, sizable populations of P. australis, as well as other likely toxin producers, P. pungens f. multiseries and P. pseudodelicatissima, have occurred biannually in Monterey Bay. Using the highly sensitive FMOC-HPLC method, we detected domoic acid whenever Fseudonitzschia australis was found in the plankton, even at densities as low as 4.0 × 103 cells/L. Based on correlations of domoic acid and P. australis abundances and the overwhelming biovolume dominance of P. australis, we conclude that P. australis has been the major domoic acid producer during the period of our study. Our study suggests that P. australis cells may always be toxic in natural populations and that toxin concentrations on a per-cell basis have no statistically significant relationship to population density or to nutrient concentrations other than silicate. Cellular levels of domoic acid were positively correlated with silicate concentrations, which is at variance with reports from prior culture experiments. These conclusions must be tentative because of the limited extent of our sampling. Nevertheless, these preliminary data indicate that further investigations of environmental conditions affiliated with cell growth and toxin production in P. australis are warranted. As a practical matter, domoic acid in the pelagic environment cannot be reliably or consistently detected by monitoring domoic acid levels in intertidal mussels. Direct measurement of domoic acid using sensitive HPLC methods is probably the most cost-effective and accurate approach for an ongoing phycotoxin monitoring program. © 1994 wiley-Liss, Inc.

Near-surface waters ranging from the Pacific subarctic (58°N) to the Southern Ocean (66°S) contain the neurotoxin domoic acid (DA), associated with the diatom Pseudo-nitzschia . Of the 35 stations sampled, including ones from historic iron fertilization experiments (SOFeX, IronEx II), we found Pseudo-nitzschia at 34 stations and DA measurable at 14 of the 26 stations analyzed for DA. Toxin ranged from 0.3 fg·cell −1 to 2 pg·cell −1 , comparable with levels found in similar-sized cells from coastal waters. In the western subarctic, descent of intact Pseudo-nitzschia likely delivered significant amounts of toxin (up to 4 μg of DA·m −2 ·d −1 ) to underlying mesopelagic waters (150–500 m). By reexamining phytoplankton samples from SOFeX and IronEx II, we found substantial amounts of DA associated with Pseudo-nitzschia . Indeed, at SOFeX in the Antarctic Pacific, DA reached 220 ng·L −1 , levels at which animal mortalities have occurred on continental shelves. Iron ocean fertilization also occurs naturally and may have promoted blooms of these ubiquitous algae over previous glacial cycles during deposition of iron-rich aerosols. Thus, the neurotoxin DA occurs both in coastal and oceanic waters, and its concentration, associated with changes in Pseudo-nitzschia abundance, likely varies naturally with climate cycles, as well as with artificial iron fertilization. Given that iron fertilization in iron-depleted regions of the sea has been proposed to enhance phytoplankton growth and, thereby, both reduce atmospheric CO 2 and moderate ocean acidification in surface waters, consideration of the potentially serious ecosystem impacts associated with DA is prudent.

Domoic acid (DA) is a neurotoxin that causes amnesic shellfish poisoning, and fish are recog- nized vectors of DA to marine fauna. However, the exposure of anglers through consumption of DA- contaminated fish is unknown. We measured DA in 11 fish species targeted by Santa Cruz Wharf (SCW) anglers in Monterey Bay, California, USA, and sur- veyed anglers regarding their fish consumption pat- terns. In addition, we used California mussel Mytilus californianus DA data provided by the state of Califor- nia and our measurements of DA in seawater to exam- ine the associations between DA in fish viscera versus in mussels and seawater. DA was detected in the vis- cera of 7 fish species commonly consumed by anglers, and toxin uptake in fishes varied according to their diet. DA was almost entirely in the viscera, with low DA concentrations detected in muscle tissue. The majority of anglers (58% of 565) reported consuming their catch, with a small fraction ingesting the viscera. Total DA concentrations in fish decreased significantly after 11 mo storage at -20°C. DA concentration in sea- water and California mussels was correlated with DA in the viscera of some but not all fish groups. We con- clude that SCW anglers who consume their catch are exposed to asymptomatic DA doses, and that exposure is a function of the species and parts consumed, as well as storage methods and DA levels in the seawater when the fish are caught.

Harmful algal blooms are increasing worldwide, including those of Pseudo-nitzschia spp. producing domoic acid off the California coast. This neurotoxin was first shown to cause mortality of marine mammals in 1998. A decade of monitoring California sea lion ( Zalophus californianus ) health since then has indicated that changes in the symptomatology and epidemiology of domoic acid toxicosis in this species are associated with the increase in toxigenic blooms. Two separate clinical syndromes now exist: acute domoic acid toxicosis as has been previously documented, and a second novel neurological syndrome characterized by epilepsy described here associated with chronic consequences of previous sub-lethal exposure to the toxin. This study indicates that domoic acid causes chronic damage to California sea lions and that these health effects are increasing.

Domoic acid (DA) is a potent food web transferred algal toxin that has caused dramatic mortality events involving sea birds and sea lions. Although no confirmed DA toxicity events have been reported in whales, here we present data demonstrating that humpback and blue whales are exposed to the toxin and consume DA contaminated prey. Whale fecal samples were found to contain DA at levels ranging from 10 to 207microg DA g(-1) feces via HPLC-UV methods. SEM analysis of whale feces containing DA, collected from krill-feeding whales, revealed the presence of diatom frustules identified as Pseudo-nitzschia australis, a known DA producer. Humpback whales were observed feeding on anchovies and sardines that contained DA at levels ranging from 75 to 444microg DA g(-1) viscera. DA contamination of whale feces and fish occurred only during blooms of toxic Pseudo-nitzschia. Additionally, several novel fish species collected during a toxic diatom bloom were tested for DA. Fish as diverse as benthic sanddabs and pelagic albacore were found to contain the neurotoxin, suggesting that DA permeates benthic as well as pelagic communities.

Over 400 California sea lions (Zalophus californianus) died and many others displayed signs of neurological dysfunction along the central California coast during May and June 1998. A bloom of Pseudo-nitzschia australis (diatom) was observed in the Monterey Bay region during the same period. This bloom was associated with production of domoic acid (DA), a neurotoxin1 that was also detected in planktivorous fish, including the northern anchovy (Engraulis mordax), and in sea lion body fluids. These and other concurrent observations demonstrate the trophic transfer of DA resulting in marine mammal mortality. In contrast to fish, blue mussels (Mytilus edulus) collected during the DA outbreak contained no DA or only trace amounts. Such findings reveal that monitoring of mussel toxicity alone does not necessarily provide adequate warning of DA entering the food web at levels sufficient to harm marine wildlife and perhaps humans.

The neurotoxin domoic acid (DA) was identified as the probable cause of death and neurological illness for approximately 70 California sea lions, Zalophus californianus, along the central California coast in May 1998. The apparent vector for sea lion intoxication was the filter-feeding northern anchovy, Engraulis mordax, via transfer of DA from the diatom, Pseudo-nitzschia australis, a natural source of this toxin. Both DA and siliceous frustules of P. australis were found in anchovy gut contents and in fecal material collected from sea lions exhibiting seizures. Anchovies contained 200.29±3.34 μg DA/g in viscera samples, 49.69±0.79 μg DA/g in whole body samples and 35.14±0.47 μg DA/g in body tissue samples. DA concentrations in sea lion fecal material ranged from 1.31 to 182.01 μg DA/g. This is the first documented case of DA toxicity in a marine mammal species.

The fecal pellets of zooplankton are thought to be major carriers of organic matter from surface to deeper waters of the oceans. As the pellets descend, they release soluble components, partially due to breakdown by associated microorganisms. Previous laboratory work of other investigators has suggested that the surface of a fecal pellet rapidly acquires bacteria, which increase in abundance until they and their protozoan consumers disrupt the pellet membrane, spilling contents into the water. In contrast, our field collections of fecal pellets from free-floating particle interceptor traps (from the “Vertex” project off Central California in 1980 and off Mexico in 1981), suggest that microbial decomposition probably is initiated in the sea from inside the fecal pellets. Transmission and scanning electron microscopy indicate that bacterial populations are most abundant in the interior of fecal pellets obtained from the sea, but that the same pellets will acquire the surface bacterial lawn typically observed in laboratory studies if maintained aboard ship. If the fecal pellets are decomposed from the inside, then the principal agents are enteric bacteria or ingested, digestion-resistant bacteria, or both. Such bacteria may differ metabolically from those that colonize the fecal pellet surfaces. Further-more, the abundance of healthy-appearing bacteria inside the pellets suggests that their metabolic activities may produce microhabitats of reduced oxygen tension that could differ considerably from that of the pellet exteriors. Decomposition in these semi-enclosed microenvironments may proceed in a manner not yet predicted by models that attribute decomposition to well-aerated, surface-dwelling bacterial populations on fecal pellets in the sea.

Nitrogen fixation, apparently by bacterial endosymbionts, is associated with intertwining chains of two species of the diatom Rhizosolenia. In situ fixation rates were enhanced by incubation in the dark, whereas concurrent shipboard experiments either underestimated or did not detect nitrogen fixation. This is the first example of nitrogen fixation associated with a bacteria-diatom symbiosis in the pelagic zone, and it indicates that these systems may contribute a significant amount of "new" nitrogen to oligotrophic waters.

Evidence of in situ metal (iron and manganese) deposition onto bacteria associated with rapidly sinking particles in the open ocean is reported. Below 100 meters, bacteria are found with extracellular capsules containing metal precipitates; the frequency of these capsules increases with depth. The capsular metal deposits appear to contribute a major portion of the weakly bound fraction of the particulate iron flux.