Reproductive potential of crimson-spotted rainbowfish (Melanotaenia fluviatilis) following short-term exposure to bass strait crude oil and dispersed crude oil

An examination of the potential of crude oil and mixtures of dispersants and crude oil to act as reproductive toxicants is reported in this article. The short-term effects of a water-accommodated fraction of crude oil (WAF) and a dispersed crude oil water-accommodated fraction (DCWAF) on selected reproductive end points were measured by conducting 3-day exposures to the crimson-spotted rainbowfish (Melanotaenia fluviatilis). Exposures were followed by 14-day depuration periods to determine the ability of fish to recover from the exposure. There were no changes to egg production, hatchability, or larval lengths for the WAF and DCWAF test periods. There were no changes to plasma estradiol or testosterone concentrations, gonadosomatic indices, or histopathological organization of gonad tissues after the exposure and depuration periods for both WAF and DCWAF. As reproductive parameters were not altered after 3 days of exposure and 14 days of depuration, crimson-spotted rainbowfish were able to endure short-term exposures to crude oil and dispersed crude oil.     

Heterosigma akashiwo in Patagonian Fjords: Genetics,Growth, Pigment Signature and Role of PUFA and ROS in Ichthyotoxicity

Heterosigma akashiwo is the only raphidophyte described for Chilean waters. A recent 2021 fish-killing bloom event of this raphidophyte ignited scientific research, but the ichthyotoxic mechanism and environmental conditions that promote its growth are still unclear. This is the first study confirming the occurrence of H. akashiwo in Chilean waters on the basis of the region D1/D2 of the 28S ribosomal gene. The pigment signature of the CREAN_HA03 strain revealed chlorophyll-a, fucoxanthin, and violaxanthin as the most abundant pigments, but profiles were variable depending on culture and field conditions. A factorial temperature–salinity growth experiment showed a maximal growth rate of 0.48 d⁻¹ at 17 °C and 35 in salinity, but reached a maximal cell abundance of ~50,000 cells mL⁻¹ at 12 °C and 25 in salinity. The fatty acid profile included high levels of saturated (16:0) and polyunsaturated (18:4 ω3; 20:5 ω3) fatty acids, but superoxide production in this strain was low (~0.3 pmol O^2– cell⁻¹ h⁻¹). The RTgill-W1 bioassay showed that the H. akashiwo strain was cytotoxic only at high cell concentrations (>47,000 cells mL⁻¹) and after cell rupture. In conclusion, salmon mortality during H. akashiwo bloom events in Patagonian fjords is likely explained by the high production of long-chain PUFAs at high cell densities, but only in the presence of high ROS production.     

Confirmation of brevetoxin metabolism in cockle, Austrovenus stutchburyi, and greenshell mussel, Perna canaliculus, associated with New Zealand neurotoxic shellfish poisoning, by controlled exposure to Karenia brevis culture.

We examined metabolism of PbTxs in New Zealand cockle, Austrovenus (A.) stutchburyi, and greenshell mussel, Perna (P.) canaliculus, by means of liquid chromatography coupled with tandem mass spectrometry. PbTx-2, PbTx-3 and BTX-B5 were detected in Karenia (K.) brevis culture medium in the ratio of ca. 50:2:5. The amounts of PbTx-3 and BTX-B5 were greatly increased in both seawater and shellfish exposed to K. brevis cultures or supernatant prepared by disruption of K. brevis under appropriate condition, while those of PbTx-2 were decreased. Some PbTx-2 was present in P. canaliculus, but not in A. stutchburyi. Low levels of BTX-B1 were detected in A. stutchburyi, but not P. canaliculus. Levels of PbTx-3 and BTX-B5 were highest immediately after exposure and then declined rapidly in both shellfish. BTX-B1 increased in concentration after exposure, and was then gradually eliminated from A. stutchburyi. Three successive exposures of A. stutchburyi to K. brevis cultures resulted in similar initial levels of PbTx-3 and BTX-B5, while BTX-B1 accumulated after each dose. In P. canaliculus, initial levels of PbTx-3 were similar, while PbTx-2 and BTX-B5 accumulated after each dose. PbTx-3 and BTX-B5 are proposed to be suitable markers for monitoring shellfish toxicity after a red tide event.     

Acute skin phototoxicity in hairless mice following exposure to crude shale oil or natural petroleum oil

The acute skin phototoxicity in hairless mice resulting from applications of crude shale oil or natural petroleum oil and exposure to near ultraviolet light has been investigated. In addition, the absorption and excitation-emission spectra of these oils have been described. While the application of crude shale oil alone was shown to elicit acute skin damage, the combination of the crude shale oil and near ultraviolet light induced skin damage much more severe than that produced by either agent alone. In contrast, the application of the natural petroleum oil alone was completely nontoxic and the combination of natural petroleum oil and near ultraviolet light was less toxic than that of the crude shale oil in the absence of light. These results suggest that a potential health hazard exists from contact with shale oil compounds and exposure to near ultraviolet light from either the sun or artificial sources.     

Characterization of polar brevetoxin derivatives isolated from Karenia brevis cultures and natural blooms.

Several novel brevetoxin derivatives were isolated and identified in Karenia brevis cultures and natural blooms by using solid phase extraction (SPE) and LC/MS(MS) techniques. These analogs were more polar compared with previously described brevetoxins, and were poorly extractable by conventional non-polar solvent (chloroform) partitioning. Brevetoxin analogs were structurally confirmed as hydrolyzed (open A-ring) forms of brevetoxins PbTx-1, PbTx-7, PbTx-2, and PbTx-3, and of oxidized PbTx-1 and PbTx-2. Some of these open A-ring derivatives were in greater abundance than their non-hydrolyzed counterparts. All were in much greater abundance in bloom water filtrate compared with cell-rich fractions. Open A-ring compounds were cytotoxic in mouse neuroblastoma (N2a) cell assay. In the K. brevis bloom-exposed Eastern oyster, brevetoxin metabolites with opened A rings were identified (e.g., open-ring cysteine-PbTx conjugates), contributing to their overall toxin burden.     

Monitoring of brevetoxins in the Karenia brevis bloom-exposed Eastern oyster (Crassostrea virginica).

Brevetoxin uptake and elimination were examined in Eastern oyster (Crassostrea virginica) exposed to recurring blooms of the marine alga Karenia brevis in Sarasota Bay, FL, over a three-year period. Brevetoxins were monitored by in vitro assays (ELISA, cytotoxicity assay, and receptor binding assay) and LC-MS, with in vivo toxicity of shellfish extracts assessed by the traditional mouse bioassay. Measurements by all methods reflected well the progression and magnitude of the blooms. Highest levels recorded by mouse bioassay at bloom peak were 157 MU/100g. Oysters were toxic by mouse bioassay at levels >or=20 MU/100g for up to two weeks after bloom dissipation, whereas brevetoxins were measurable by in vitro assays and LC-MS for several months afterwards. For the structure-based methods, summed values for the principal brevetoxin metabolites of PbTx-2 (cysteine and cysteine sulfoxide conjugates), as determined by LC-MS, were highly correlated (r(2)=0.90) with composite toxin measurements by ELISA. ELISA and LC-MS values also correlated well (r(2)=0.74 and 0.73, respectively) with those of mouse bioassay. Pharmacology-based cytotoxicity and receptor binding assays did not correlate as well (r(2)=0.65), and were weakly correlated with mouse bioassay (r(2)=0.48 and 0.50, respectively). ELISA and LC-MS methods offer rapid screening and confirmation, respectively, of brevetoxin contamination in the oyster, and are excellent alternatives to mouse bioassay for assessing oyster toxicity following K. brevis blooms.     

Brevetoxin metabolism and elimination in the Eastern oyster (Crassostrea virginica) after controlled exposures to Karenia brevis.

The metabolism and elimination of brevetoxins were examined in the Eastern oyster (Crassostrea virginica) following controlled exposures to Karenia brevis cultures in the laboratory. After a 2-day exposure period ( approximately 62 million cells/oyster), elimination of brevetoxins and their metabolites was monitored by using liquid chromatography/mass spectrometry (LC/MS). Composite toxin in oyster extracts was measured by in vitro assay (i.e. cytotoxicity, receptor binding, and ELISA). Of the parent algal toxins, PbTx-1 and PbTx-2 were not detectable by LC/MS in K. brevis-exposed oysters. PbTx-3 and PbTx-9, which are accumulated directly from K. brevis and through metabolic reduction of PbTx-2 in the oyster, were at levels initially (after exposure) of 0.74 and 0.49 microg equiv./g, respectively, and were eliminated largely within 2 weeks after dosing. PbTx-7 and PbTx-10, the reduced forms of PbTx-1, were non-detectable. Conjugative brevetoxin metabolites identified previously in field-exposed oysters were confirmed in the laboratory-exposed oysters. Cysteine conjugates of PbTx-1 and PbTx-2, and their sulfoxides, were in the highest abundance, as apparent in LC/MS ion traces, and were detectable for up to 6 months after dosing. Composite toxin measurements by in vitro assay also reflected persistence (up to 6 months) of brevetoxin residues in the oyster. Levels of cysteine conjugates, as determined by LC/MS, were well correlated with those of composite toxin, as measured by ELISA, throughout depuration. Composite toxin levels by cytotoxicity assay were well correlated with those by receptor binding assay. Cysteine-PbTx conjugates are useful LC/MS determinants of brevetoxin exposure and potential markers for composite toxin in the Eastern oyster.     

Induction of microsomal mixed-function oxidase system components in striped mullet by short-term exposure to crude oil

Juvenile striped mullet were exposed to slicks of either Empire Mix (of American origin) or Saudi Arabian crude oils for 1 or 3 weeks. Microsomal protein was elevated. Cytochromes P-450 and b₅ and NADPH-cytochrome c and NADPH-dichlorophenolindophenol reductases were induced but NADH-cytochrome c and NADH-cytochrome b₅ reductases were not. Induction by Empire Mix oil increased with time of exposure but induction by Saudi Arabian oil remained about the same with time. The induction of microsomal components was greater for Saudi Arabian oil than for Empire Mix oil at one week of exposure, but the reverse was true at three weeks.     

Variation in brevetoxin and brevenal content among clonal cultures of Karenia brevis may influence bloom toxicity

Karenia brevis, the major harmful algal (HA) species in the Gulf of Mexico, produces a suite of brevetoxins and brevenal, a nontoxic brevetoxin antagonist. K. brevis growth is reported to be optimum at oceanic conditions, yet blooms are most problematic in coastal waters. Differences in growth rate, total brevetoxin production, brevetoxin profiles and brevenal production were evaluated among eight K. brevis clones grown at salinities of 35 and 27, but otherwise identical conditions. All measured parameters varied significantly among clones and the individual responses to decreased salinity varied as well. At 27, growth rates of four clones increased (Wilson, TXB3, SP1 and SP2), but decreased in three others (TXB4, SP3 and NBK) as compared to 35. Total brevetoxin cellular concentration varied up to ～ten-fold among clones. For most clones (5 of 8), no significant difference in total toxin production between salinity treatments was observed; however, there was a shift in brevetoxin profiles to a higher proportion of PbTx-1 vs. PbTx-2 (in 7 of 8 clones). Brevenal production decreased in the majority of the clones (6 of 8) when grown at a salinity of 27. Results suggest that K. brevis produces more PbTx-1 and less brevenal in lower salinity waters.     

Immunotoxic effects of in vitro exposure of dolphin lymphocytes to Louisiana sweet crude oil and Corexit™

The Deepwater Horizon oil spill was one of the worst environmental disasters on record in the United States. Response efforts to reduce the magnitude of the oil slick included the use of thousands of gallons of the chemical dispersant Corexit™ in surface and deep‐water environments. The immunotoxicity of Louisiana sweet crude oil and the chemical dispersant Corexit was examined using lymphocyte proliferation (LP) and natural killer cell (NK) assays as measures of impact on the adaptive (LP) and innate (NK) immune response in bottlenose dolphins. Study results show that both high‐energy media‐accommodated fractions (MAF) and chemically enhanced MAF (CEMAF) mixtures modulate immune function. Following exposure to Louisiana sweet crude, both B‐ and T‐cell proliferation of white blood cells was increased for all exposure concentrations, compared to control; however, this increase was only significant for the 50% and 100% treatments. In contrast, exposure of white blood cells to the CEMAF mixture significantly decreased both T‐ and B‐cell proliferation in the 25%, 50% and 100% treatments. NK cell activity was enhanced significantly by CEMAF mixtures for the 50% and 100% treatments. The immunosuppression of LP at environmentally relevant concentrations of oil and dispersant suggests that marine mammals may be unable to mount an adequate defense against xenobiotic threats following exposure to oil and dispersant, leaving them more susceptible to disease. In contrast, NK cell activity was significantly enhanced, which may increase an organism's tumor or viral surveillance ability by mounting an enhanced immune response. Copyright © 2016 John Wiley & Sons, Ltd.     

Effects of the dinoflagellate Karenia brevis on larval development in three species of bivalve mollusc from Florida.

The effects of Karenia brevis (Wilson clone) on larval survival and development of the northern quahog, Mercenaria mercenaria, eastern oyster, Crassostrea virginica and bay scallop, Argopecten irradians, were studied in the laboratory. Larvae were exposed to cultures of whole and lysed cells, with mean total brevetoxin concentrations of 53.8 and 68.9 microgL(-1), respectively. Survival of early (3-day-old) larvae was generally over 85% for all shellfish species at K. brevis densities of 100 cells ml(-1) or less, and not significantly different between whole and lysed culture. At 1000 cells ml(-1), survival was significantly less in lysed culture than whole culture for both M. mercenaria and C. virginica. Survival of late (7-day-old) larvae in all three species was not significantly affected by K. brevis densities of 1000 cells ml(-1) or less. At 5000 cells ml(-1), however, survival was reduced to 37%, 26% and 19% for A. irradians, M. mercenaria and C. virginica, respectively. Development of C. virginica and M. mercenaria larvae was protracted at K. brevis densities of 1000 cells ml(-1). These results suggest that blooms of K. brevis, and particularly their associated brevetoxins, may have detrimental consequences for Florida's shellfisheries by disrupting critical larval processes. Special attention should be paid to blooms of K. brevis where these shellfish occur naturally or where aquaculture and restoration activities are either ongoing or planned.     

Toxicity of crude oil and dispersed crude oil to ghost shrimp Palaemon serenus and larvae of Australian bass Macquaria novemaculeata

Acute 96‐h LC50 values of the water‐accommodated fraction (WAF) of crude oil, dispersants (Corexit 9500 and Corexit 9527) and dispersed oil combinations were determined in semistatic bioassays with seawater, using the ghost shrimp Palaemon serenus and larval Australian bass (fish) Macquaria novemaculeata. Sodium dodecyl sulphate (SDS) and zinc sulphate were used as reference toxicants and identical bioassays were conducted using these compounds. Total petroleum hydrocarbon (TPH) uptake of shrimp was also measured on the samples taken from the bioassays. The nominal mean (n=4) 96‐h LC50 standard error (SE) values for WAF of crude oil, Corexit 9527, Corexit 9500, dispersed oil (9527) and dispersed oil (9500) were 258,000 ppm (13,000), 49.4 ppm (6.4), 83.1 ppm (5.8), 8.1 ppm (0.3), and 3.6 ppm (0.3) in the shrimp bioassays, respectively. The nominal mean (n=4) 96‐h LC50 (SE) values calculated from the fish larval bioassays were 465,000 ppm (16,000), 14.3 ppm (0.9), 19.8 ppm (1.6), 28.5 ppm (1.4), and 14.1 ppm (2.6) for WAF of crude oil, Corexit 9527, Corexit 9500, dispersed oil (9527), and dispersed oil (9500), respectively. These LC50 values indicate that dispersed oil combinations were significantly more toxic to these organisms than WAF of crude oil. TPH uptake of shrimp increased in correlation to exposure concentrations, and the presence of dispersant made oil more available for shrimp. © 2000 John Wiley & Sons, Inc. Environ Toxicol 15: 91–98, 2000     

Confirmation of brevetoxin metabolism in the Eastern oyster (Crassostrea virginica) by controlled exposures to pure toxins and to Karenia brevis cultures.

Previously, we analyzed Eastern oysters (Crassostrea virginica) naturally exposed to a Karenia brevis red tide and found that brevetoxins (PbTx) are rapidly accumulated and metabolized. Several metabolites were isolated and later identified, including a cysteine-PbTx conjugate (MH(+): m/z 1018) and its sulfoxide product (m/z 1034). In the present study, we confirm and extend those findings by examining PbTx metabolism and elimination in oysters exposed to pure toxins (PbTx-2 and -3) under controlled conditions. Waterborne PbTx-3 was rapidly accumulated, but not metabolized, in the oyster and was largely eliminated within 2 weeks after exposure. In contrast, PbTx-2 was accumulated and rapidly metabolized. Metabolites of PbTx-2 included the reduction product PbTx-3 (m/z 897), and the cysteine conjugates (m/z 1018 and 1034) isolated previously from the field samples. Levels of the metabolite PbTx-3 in PbTx-2-exposed oysters were highest immediately after exposure and declined at a rate similar to parent PbTx-3 in PbTx-3-exposed oysters. Cysteine-PbTx persisted for 8 weeks after exposure. The same metabolites were confirmed in oysters exposed to laboratory cultures of K. brevis. PbTx metabolites contribute to neurotoxic shellfish poisoning (NSP) and should be included in analytical protocols for monitoring shellfish toxicity after a K. brevis red tide event.     

Studies of the effect of Psi-APONIN from Nannochloris sp. on the Florida red tide organism Karenia brevis.

Studies were conducted on the conditions under which the red tide organism, Karenia brevis (a.k.a., Gymnodinium breve), was treated with Nannochloris sp. The latter organism is known to produce cytolytic agents called Apparent Oceanic Naturally Occurring Cytolin (APONINs). Conventional wisdom might suggest that brevetoxins would be released upon destruction of the single-celled dinoflagellate K. brevis and that efforts to treat red tide outbreaks would lead to release of brevetoxins and enhanced toxicity toward marine species. Earlier studies described conditions by which K. brevis cells were converted to a non-motile form when cultures of K. brevis were treated with an isolate (Psi-APONIN) produced by Nannochloris sp. but when centrifuged only a small amount of the toxin was released. The present study confirms that the toxin is not released when the K. brevis is undisturbed, however, when the culture is stressed (stirred with a magnetic stirring bar for 24, 48, and 72h) toxin was released, and the toxicity could be measured using a Microtox analyzer. In the study, it was found that at as few as eighty cells of K. brevis produced a toxic effect of 20% as measured by the effect on Vibrio fischeri. Nannochloris sp. had no effect on the bacteria used in the Microtox analyzer, nor did interaction of Nannochloris sp. with K. brevis in the short term. This effect is presumed to be due to the production of Psi-APONIN and conversion of K. brevis to a non-motile or resting form.     

Toxicity of French strains of the dinoflagellate Prorocentrum minimum experimental and natural contaminations of mussels.

Mediterranean strains of Prorocentrum minimum do not appear to have the same toxic component as Japanese strains since they showed no cytotoxicity for hepatocytes in culture. However, their toxic components, which appear to block calcium channels, were detectable by the immobilisation test on Diptera larvae. A bio-accumulation experiment in the laboratory showed that the toxins could accumulate in nearly equivalent amounts in the hepatopancreas and meat of cultured mussels. The same toxicity was found in natural samples collected in a period of bloom of P. minimum. These results suggest that P. minimum could be responsible for shellfish toxicity in the natural environment and thus present a risk for human health.     

Comparative effects of the toxic dinoflagellate Karenia brevis on clearance rates in juveniles of four bivalve molluscs from Florida, USA.

The effects of Karenia brevis (Gymnodiniales, Gymnodiniaceae) on the feeding activity of juveniles of four species of bivalve mollusc were examined in the laboratory to assess the potential impacts on these important shellfish populations from Florida. Clearance rates were determined under short-term (one hour) static and long-term (two days) flow-through conditions using both whole and lysed cultures of K. brevis. Under short-term conditions, the bay scallop, Argopecten irradians, was the most sensitive species, exhibiting a 79% reduction in clearance rate at 1000 cells ml(-1) of whole K. brevis culture compared to the control (no K. brevis). The eastern oyster, Crassostrea virginica, was the least responsive, showing a 38% reduction in clearance rate between the same treatments. The green mussel, Perna viridis, and the northern quahog, Mercenaria mercenaria, displayed intermediate responses. Similar results were also observed during long-term exposures to a continuous supply of K. brevis. Bay scallops showed a significant decline in clearance rate at 100 cells ml(-1) after 24h exposure; clearance rate of oysters was not affected by K. brevis at this concentration. No mortality was observed for any species during these brief exposures. The prospect for recovery of bay scallop populations in Florida estuaries where they were once abundant may be hampered by recurring blooms of K. brevis. Reduced clearance rates in M. mercenaria at high K. brevis densities could translate into poor growth of cultured Florida hard clams. On the other hand, P. viridis, which also showed reduced clearance rates at high K. brevis concentrations, might be negatively impacted by K. brevis blooms, thereby affecting their ability to spread into estuaries hampered by recurring toxic algal blooms.     

Responses of hepatocytes to DDT and methyl mercury exposure

The aim of the current work was to investigate the effects of dichlorodiphenyltrichloroethane (DDT) and monomethyl mercury (MeHg) on hepatocytes from tropical fish Hypostomus commersoni (cascudo). In order to verify DDT and MeHg impacts on the redox milieu, cells were exposed for 4 days to 50 nM of DDT, 0.25 and 2.5 μM of MeHg and to a combination of 50 nM of DDT and 0.25 μM of MeHg. These concentrations were compared with those previously published (Filipak Neto et al., 2008) for the predator fish Hoplias malabaricus (traíra). The effects were mostly noticeable on reduced glutathione concentration and δ-aminolevulinic acid dehydratase and glutathione S-transferase activity. Catalase activity increased in the group exposed to 2.5 μM of MeHg and hydrogen peroxide levels decreased in all exposed groups. Also, superoxide anion levels decreased in the groups exposed to 2.5 μM of MeHg and DDT1MeHg group. Cell viability decreased only in the DDT exposed group, demonstrating that the antioxidant defense mechanism of H. commersoni hepatocytes is more efficient than H. malabaricus. These results corroborate the resistance of H. commersoni to polluted areas and support the hypothesis that this species is more resistant to DDT and MeHg than H. malabaricus species.     

First Detection of Tetrodotoxin in Greek Shellfish by UPLC-MS/MS Potentially Linked to the Presence of the Dinoflagellate Prorocentrum minimum

During official shellfish control for the presence of marine biotoxins in Greece in year 2012, a series of unexplained positive mouse bioassays (MBA) for lipophilic toxins with nervous symptomatology prior to mice death was observed in mussels from Vistonikos Bay–Lagos, Rodopi. This atypical toxicity coincided with (a) absence or low levels of regulated and some non-regulated toxins in mussels and (b) the simultaneous presence of the potentially toxic microalgal species Prorocentrum minimum at levels up to 1.89 × 10³ cells/L in the area’s seawater. Further analyses by different MBA protocols indicated that the unknown toxin was hydrophilic, whereas UPLC-MS/MS analyses revealed the presence of tetrodotoxins (TTXs) at levels up to 222.9 μg/kg. Reviewing of official control data from previous years (2006–2012) identified a number of sample cases with atypical positive to asymptomatic negative MBAs for lipophilic toxins in different Greek production areas, coinciding with periods of P. minimum blooms. UPLC-MS/MS analysis of retained sub-samples from these cases revealed that TTXs were already present in Greek shellfish since 2006, in concentrations ranging between 61.0 and 194.7 μg/kg. To our knowledge, this is the earliest reported detection of TTXs in European bivalve shellfish, while it is also the first work to indicate a possible link between presence of the toxic dinoflagellate P. minimum in seawater and that of TTXs in bivalves. Confirmed presence of TTX, a very heat-stable toxin, in filter-feeding mollusks of the Mediterranean Sea, even at lower levels to those inducing symptomatology to humans, indicates that this emerging risk should be seriously taken into account by the EU to protect the health of shellfish consumers.     

The Effects of the Harmful Algal Bloom Species Karenia brevis on Survival of Red Porgy (Pagrus pagrus) Larvae

The harmful algal bloom species, Karenia brevis, forms annual, often intense blooms in the Gulf of Mexico, particularly along the west Florida shelf. Though the ability of K. brevis blooms to cause mass mortalities in juvenile fish are well documented, the direct effect of bloom concentrations on larval fish has not been studied extensively. To better understand the potential effect of K. brevis on larval fish survival, laboratory spawned red porgy (Pagrus pagrus) larvae from 4–26 days post-hatch were exposed to concentrations of K. brevis observed in the field for either 24 or 48 h. This species is representative of fish which spawn in regions of the Gulf of Mexico and whose larvae are epipelagic and may encounter K. brevis blooms. In this study, three different K. brevis strains varying in the amount of brevetoxin produced were tested. Larval survivorship was found to be inversely proportional to the amount of brevetoxin produced by each strain. The EC₅₀ value from the combined 24 h experiments was ~163,000 K. brevis cells L⁻¹, which corresponds to cell concentrations found in moderately dense blooms. Larval mortality also increased substantially in the 48 h versus 24 h exposure treatments. These findings indicate K. brevis blooms have the potential to contribute to natural mortality of fish larvae and further reduce inter-annual recruitment of fishery species whose stocks in the Gulf of Mexico may already be depleted.