Relationship between blood parameters and biometric indices of Sparus aurata and Dicentrarcus labrax cultured in onshore tanks

The aim of this study was to evaluate the possible correlation between biometric indices (length and weight) and some blood parameters in 225 Gilthead sea bream (Sparus aurata) and 225 European sea bass (Dicentrarchus labrax) cultured in onshore tanks with the same physical–chemical conditions of water (temperature, soluble oxygen and salinity). From each fish, blood samples were collected to evaluate Red Blood Cell Count, Haematocrit, Haemoglobin Concentration, White Blood Cell Count, Thrombocyte Count, Mean Corpuscular Volume, Mean Corpuscular Haemoglobin and Mean Corpuscular Haemoglobin Concentration, glucose, lactate and Total Proteins. In both species, the relationship between the biometric indices and blood parameters was evaluated using linear regression analysis that showed a statistically significant correlation between the biometric indices and seven of the studied parameters. These results showed that fish size is an important factor in the interpretation of haematological and biochemical profiles in order to evaluate correctly the health status of the fish.     

Biometric relationships between body and otolith measurements in nine demersal fishes from north‐eastern Tasmanian waters,Australia

Relationships between fish and otolith measurements were analyzed in nine demersal fishes from the north‐eastern Tasmanian waters: Foetorepus calauropomus, Trachurus declivis, Parequula melbournensis, Neosebastes scorpaenoides, Platycephalus aurimaculatus, Platycephalus bassensis, Platycephalus conatus, Lepidotrigla mulhalli and Lepidotrigla vanessa. The values of exponent b from the relationships between fish weight and fish total length, total length and otolith length, total length and otolith width, and fish weight and otolith length were estimated. All above relationships were statistically significant. The analysis of covariance (ANCOVA) was used to test the effect of the categorical factor of species in the fish length and otolith length relationship. Significant differences in fish length and otolith length relationship between species were highlighted in both gurnards and flatheads. Results from this study will provide novel information on quantitative biometric relationships between body and otolith measurements of fish species in Australian waters.     

Biomarkers of toxicity in Clarias gariepinus exposed to sublethal concentrations of polycyclic aromatic hydrocarbons

Physiological, biochemical and histological indices in Clarias gariepinus broodstock, and teratogenic indices in embryos exposed to sublethal concentrations of naphthalene, phenanthrene and pyrene were investigated in 2014 using a static-renewal bioassay protocol. Phenanthrene (1.41 mg l^?1) was the most toxic, followed by pyrene (1.53 mg l^?1) and naphthalene (7.21 mg l^?1), based on 96 h LC50 values. Hepatosomatic indices were significantly higher in naphthalene- and pyrene-treated males compared with solvent controls, whereas fecundity in females was significantly lower by factors of 2.4 (naphthalene), 2.8 (phenanthrene) and 2.4 (pyrene), compared with controls. Catalase levels were lower in female phenanthrene-treated fish compared with controls. Histological alterations observed in PAH-treated fish include oedema, inflammatory cells, epithelial lifting and hyperplasia in the gills, vacuolation, haemosiderin pigments and sinusoidal congestion in the liver, and degenerated zona radiata in the ovary. Teratogenic effects were not observed, as evidenced by the lack of histological alterations in embryos spawned from pre-exposed broodstock. Sex-specific responses and the utility of biomarkers at cellular and individual levels of organisation are therefore demonstrated for holistic evaluations of polycyclic aromatic hydrocarbons in ecotoxicological studies.     

Visceral Adipose Tissue in Women: Longitudinal Study of the Effects of Fat Gain,Time, and Race

Objective: To determine the effects of fat gain, time, and race on the accumulation of visceral adipose tissue (VAT) in a group of normal‐weight premenopausal women. Research Methods and Procedures: Sixty‐five women participated in the study (32 African American and 33 white). The mean age of subjects was 34 ± 6 years (range, 22 to 47 years). Eligible subjects were women who had body mass indices <25 kg/m² at baseline and who had completed evaluations at baseline and at follow‐up year 1, without intervention. A subset of subjects was reevaluated annually for up to 4 years. Body composition was assessed by DXA, and VAT was determined from a single computed tomography scan. A linear mixed model was used to examine changes in VAT over time, with total body fat as a covariate Results: Total fat mass was not significantly different between races at baseline and increased significantly in both groups over time (p < 0.001). Time‐related increases in total body fat were greater in African‐American women (p < 0.01). VAT was significantly higher in white women at baseline (p < 0.01) and increased significantly over time in both races (p < 0.01), but remained higher in white women (p < 0.001). Increases in VAT, relative to total body fat, were greater than the increases in total body fat over time, independent of age and race (p < 0.001). Discussion: Gaining total body‐fat mass results in a higher increase in VAT, relative to total body fat, regardless of race and age, although African‐American women maintain a lower VAT levels across time.     

Hemato-biochemical responses of Van fish (Alburnus tarichi Guldenstadt, 1814) during sublethal exposure to cypermethrin

Cypermethrin is a highly active type II pyrethroid that is widely used to control harmful insects. The present study was conducted to investigate the possible effects of the sublethal concentration of cypermethrin (0.05 μL/L) on the biometric, hematological, and biochemical parameters of Van fish (Alburnus tarichi Guldenstadt 1814) for 24, 48, 72, and 96 h. Behavioral and physiological changes were observed in the fish treated with cypermethrin. Hematological parameters revealed a significant decrease in the erythrocyte (RBC), hemoglobin (Hb), and hematocrit (Hct) values (p ≤ .05). Serum enzymes, aspartate aminotransferase (AST, EC 2.6.1.1), alanine aminotransferase (ALT, EC 2.6.1.2), and lactate dehydrogenase (LDH, EC 1.1.1.27) activities and serum cortisol level were significantly increased (p ≤ .05). The results showed that changes in the biometric, biochemical, and hematological values of the Van fish exposed to cypermethrin can be used as biomarkers for monitoring toxicity.     

Biochemical and toxicopathic biomarkers assessed in smallmouth bass recovered from a polychlorinated biphenyl-contaminated river

Smallmouth bass (Micropterus dolomieu) were collected to quantify the nature and prevalence of biomarker responses, including biochemical indices, toxicopathic lesions and general health indices, among fish collected from polychlorinated biphenyl (PCB)-contaminated and nearby uncontaminated reaches of the Kalamazoo River, Michigan, USA. Blood and tissue samples (gill, liver, spleen, head kidney, trunk kidney, thyroid and gonads) were collected and preserved at necropsy for biochemical and histological analyses. The body condition factor and liver somatic index were significantly lower in fish collected from the downstream, contaminated site. Plasma vitellogenin was not detected in male fish collected from either site. Liver ethoxyresorufin-O-deethylase activity and liver and spleen superoxide dismutase activity were significantly depressed in fish collected from the downstream site. Significant toxicopathic lesions such as glycogen depletion, enhanced macrophage aggregates, hepatic foci of cellular alteration (i.e. preneoplastic lesions) and neoplasia were also detected in the liver of fish collected from the downstream site. This study indicates that many of the biochemical and histopathological biomarker responses were associated with liver and body tissue PCB concentrations. Taken together, the biomarkers of exposure and effect strongly suggest that fish within the downstream site are adversely affected by PCBs and other chemical stressors.     

Adiponectin and ghrelin levels and body size in normoglycemic Filipino, African-American, and white women

Objective: Prior studies have reported ethnic differences in adiponectin and ghrelin, but few have assessed the role of body size in normoglycemic women. We compared fasting adiponectin and ghrelin concentrations in normoglycemic 40‐ to 80‐year‐old Filipino, African‐American, and white women. Methods: Participants included women from the Rancho Bernardo Study (n = 143), the University of California‐San Diego Filipino Women's Health Study (n = 136), and the Health Assessment Study of African‐American Women (n = 212). A 2‐hour oral glucose tolerance test was administered; glucose, insulin, lipid, and anthropometric measurements were obtained. Fasting adiponectin and ghrelin were measured by radioimmunoassay. Results: Whites and Filipinas had similar BMI (23.7 and 24.3 kg/m², respectively), waist girth (75.6 and 77.2 cm, respectively), and total body fat (27.4 and 28.5%, respectively); African‐Americans had significantly larger BMI (28.8 kg/m²), waist girth (86.3 cm), and body fat (39.6%, p < 0.0001). Adiponectin was lower in Filipinas (8.90 µg/mL) and African‐Americans (9.67 µg/mL) compared with whites (15.6 µg/mL, p < 0.001) after adjusting for age, homeostasis model assessment of insulin resistance (HOMA‐IR), and waist‐to‐hip ratio. Compared with whites, Filipinas (β = ?5.06, p < 0.0001) and African‐Americans (β = ?6.85, p < 0.0001) had significantly lower adiponectin levels after adjusting for age, waist‐to‐hip ratio, HOMA‐IR, triglycerides, high‐density lipoprotein (HDL) cholesterol, exercise, and alcohol use. Ghrelin was significantly lower in Filipinas compared with African‐Americans (1146.9 vs. 1412.2 pg/mL, p < 0.001), and this observation persisted in multivariable analysis (β = ?245.4, p < 0.0001). Ghrelin levels did not differ between whites (1356.9 pg/mL) and either ethnic group. Discussion: Normoglycemic Filipino and African‐American women had significantly lower adiponectin concentrations than white women, and Filipinas had lower ghrelin levels than African‐Americans, independently of body size or indices of insulin resistance or lipids.     

Exposure to elevated pCO2 alters post‐treatment diel movement patterns of largemouth bass over short time scales

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8-Oxo-7,8-dihydro-2’-deoxyguanosine,reactive oxygen species and ambulatory blood pressure in African and Caucasian men: The SABPA study

Abstract

Various studies indicate a relationship between increased oxidative stress and hypertension, resulting in increased DNA damage and consequent excretion of 8-oxo-7,8-dihydro-2’-deoxyguanosine (8-oxodG). The aim of this study was to compare urinary 8-oxodG levels in African and Caucasian men and to investigate the association between ambulatory blood pressure (BP) and pulse pressure (PP) with 8-oxodG in these groups.

We included 98 African and 92 Caucasian men in the study and determined their ambulatory BP and PP. Biochemical analyses included, urinary 8-oxodG, reactive oxygen species (ROS) (measured as serum peroxides), ferric reducing antioxidant power (FRAP), total glutathione (GSH), glutathione peroxidase (GPx) and glutathione reductase (GR) activity.

The African men had significantly higher systolic (SBP) and diastolic blood pressure (DBP) (both p < 0.001). Assessment of the oxidative stress markers indicated significantly lower 8-oxodG levels (p < 0.001) in the African group. The African men also had significantly higher ROS (p = 0.002) with concomitant lower FRAP (p < 0.001), while their GSH levels (p = 0.013) and GR activity (p < 0.001) were significantly higher. Single and partial regression analyses indicated a negative association between urinary 8-oxodG levels with SBP, DBP and PP only in African men. These associations were confirmed in multiple regression analyses (SBP: R² = 0.41; β = ?0.25; p = 0.002, DBP: R² = 0.30; β = ?0.21; p = 0.022, PP: R² = 0.30; β = ?0.19; p = 0.03).

Our results revealed significantly lower urinary 8-oxodG in African men, accompanied by a negative association with BP and PP. We propose that this may indicate a dose-response relationship in which increased oxidative stress may play a central role in the up-regulation of antioxidant defence and DNA repair mechanisms.     

The effect of dissolved oxygen on the growth of young-of-the-year winter flounder,Pseudopleuronectes americanus

Synopsis The effects of constant and diurnally fluctuating levels of dissolved oxygen on the growth of young-of-the-year winter flounder,Pseudopleuronectes americanus, were examined under controlled laboratory conditions. Fish were exposed for either 10 or 11 weeks to constant levels of 6.7 (high) and 2.2 (low) mg l^–1, and a diurnal fluctuation, ranging from 2.5 to 6.4 mg 0₂l^–1. Growth rates, calculated for both standard length and weight, for fish exposed to low and diurnally fluctuating levels were significantly reduced (p < 0.001) as compared to those for fish exposed to the high level. Growth rates of fish exposed to the high level were over twice those of fish held under low oxygen conditions. Under fluctuating conditions, fish grew at intermediate rates. Following these exposures, all fish were subsequently held at 7.2 mg Oz l^–1 for five weeks. Growth rates increased over two and a half times for fish previously exposed to the low oxygen level and were significantly (p < 0.001) higher than for the other two groups.     

Lower Serum Adiponectin Levels in African‐American Boys

Objective: To examine adiponectin, an adipocyte‐secreted hormone with anti‐inflammatory and insulin‐sensitizing effects, in relation to race or gender in younger subjects. Research Methods and Procedures: The relationship of adiponectin, quantitated by radioimmunoassay, to anthropometric and metabolic factors (fasting insulin, glucose, and leptin) and reproductive hormones was examined in 46 healthy African Americans (25 girls/21 boys) and 40 whites (20 girls/20 boys) ranging in age from 12 to 21 years. Results: There was no statistical difference in BMI or in BMI percentile among the four groups. Sums of skinfolds, but not skinfold percentile, were significantly lower in boys than girls (p = 0.001 and p = 0.896, respectively), whereas there was no difference between racial groups. Leptin was significantly greater in girls (p = 0.0002). There was no difference in fasting serum glucose, insulin, or homeostasis model assessment score among any of the groups. There was a significant negative univariate relationship between serum adiponectin and both BMI and BMI percentile for the entire group (p = 0.006 and p = 0.005). In a multivariate model, BMI percentile (p = 0.005) and the interaction between race and gender (p = 0.026) were significant predictors of serum adiponectin. In this model, African‐American boys had the lowest serum adiponectin level, 37% less than white boys, who had the highest adiponectin levels. Discussion: Serum adiponectin levels are reduced in young obese subjects (African Americans and whites) and are lower in African‐American boys than white boys. A lower adiponectin level in African‐American boys may predispose this group to a greater risk of diabetes and cardiovascular disease.     

Sociodemographics of Pet Ownership among Adolescents in Great Britain: Findings from the HBSC Study in England,Scotland, and Wales

The aim of this study was to assess the prevalence of pet ownership among adolescents in Great Britain and identify any sociodemographic differences between pet owners and non-pet owners. A total of 14,328 11-to 15-year-old adolescents from England, Scotland, and Wales were included in the analysis. Results revealed 15-year-old adolescents were significantly more likely than 11-year-old adolescents to own dogs (OR = 1.146, p < 0.001) but less likely to own fish, reptiles, or amphibians (OR = 0.629, p < 0.001), and small mammals (OR = 0.630, p < 0.001). Thirteen-year-olds were significantly more likely than 11-year-olds to own dogs (OR = 1.240, p = 0.021) and birds (OR = 1.299, p = 0.010), but significantly less likely to own fish, reptiles, or amphibians (OR = 0.795, p < 0.001). No gender differences were found. White adolescents were more likely than non-white adolescents to own all pet types. Those living in single-parent families were significantly more likely than those living with two parents to own dogs (OR = 1.186, p = 0.013) and cats (OR = 1.319, p < 0.001). Furthermore, those who reported living in stepfamilies were also more likely to own cats (OR = 1.428, p < 0.001). Adolescents with siblings were more likely to own cats (OR = 1.391, p = 0.001), fish, reptiles, or amphibians (OR = 1.220, p = 0.037) than adolescents without siblings. Adolescents with employed parents (both or one) were significantly more likely than those with unemployed parents to own dogs (OR = 1.414, p = 0.002) and birds (OR = 1.523, p = 0.018). Adolescents from high-affluence families were less likely than adolescents from low-affluence families to own dogs (OR = 0.888, p = 0.037), small mammals (OR = 0.832, p = 0.005), and birds (OR = 0.801, p = 0.046). Furthermore, family affluence differences were found in different pet types. Differences in all pet types and siblings were also found in a proxy measure of attachment to pets. This study provides evidence that pet ownership is related to several sociodemographic factors. These are relevant to take into account when performing HAI studies on adolescents.     

Protective role of supplemental vitamin E and selenium on lipid peroxidation, vitamin E, vitamin A, and some mineral concentrations of Japanese quails reared under heat stress

This study was conducted to determine the effects of vitamin E and selenium (Se) on lipid peroxidation (MDA), serum and liver concentration of antioxidant vitamins, and some minerals of Japanese quails reared under heat stress (34°C). One hundred twenty 10-d-old Japanese qualis (60 males, 60 females) were randomly assigned to 4 treatment groups, 3 replicates of 10 birds each. The experiment was designed in a 2×2 factorial arrangement using two levels of vitamin E (125 and 250 mg/kg of diet) and two levels of selenium (0.1 and 0.2 mg/kg of diet). Greater dietary vitamin E and selenium inclusions resulted in a greater (p=0.001) serum vitamin E and vitamin A, but lower (p=0.001) MDA concentrations. Liver vitamin E and vitamin A concentrations increased (p=0.001) and MDA concentrations decreased (p=0.001) when both dietary vitamin E and selenium increased. No interactions between vitamin E and selenium were detected (p≥0.11) for any parameters. Increasing both dietary vitamin E and selenium caused an increase in serum concentrations of Fe and Zn (p=0.001), but a decrease in serum concentration of Cu (p=0.001). Results of the present study showed that dietary vitamin E and selenium have synergistic effects and that supplementing a combination of dietary vitamin E (250 mg/kg of diet) and selenium (0.2 mg/kg of diet) offers a good management practice to reduce heat stress-related depression in performance of Japanese quails.     

Human-induced changes in the composition of fish communities in the African Great Lakes

Summary Large natural African lakes contain unique and diverse fish faunas which have evolved within each lake in a comparatively short period of time. members of the family Cichlidae are particularly diverse, although there is strong evidence to show that the haplochromines in Lake Victoria, and possibly Lake Malawi, are monophyletic. The unique faunas in Lakes Victoria and Kyoga have been subject to perturbations from the introduction of exotic fish, and the faunas in these and other lakes have been disturbed by fishing activities and other human endeavours.Factors governing the establishment of exotic species are not clearly understood. The exotic fish must be physiologically adapted to their new environment, able to compete successfully both for habitat and for food at each stage of their life history, able to avoid predation and must have a suitable reproductive potential. Although about 50 species of fish have been introduced into African inland waters, including reservoirs, only comparatively few, in particular Nile perch (Lates niloticus), various cichlids (especially tilapias) and clupeids (Limnothrissa miodon), have been successful in establishing themselves. Those that have become established have had obvious but unquantifiable impacts on the indigenous faunas.It is difficult to differentiate between the effects of fishing and of the presence of alien fish on the fish species composition of the lakes (Witte et al., 1992). Many of the lakes were overfished before introductions were made, with a resultant decline in some species, especially the larger ones, and the virtual disappearance of others. Some lake fish faunas, such as those of Lakes Kyoga and Victoria, which have been subjected to the perturbations described above, continue to change rapidly (Ogutu-Owayo, 1990b).There is a fundamental need to collect biological information on the fish communities of African lakes for effective management, resulting not only in the conservation of unique fish faunas but also the production of sustainable fish yields for the people relying on this source of protein. This information is required before any more introductions of exotic fish are made.     

A note on copper bioaccumulation in Mozambique tilapia,Oreochromis mossambicus (Osteichthyes: Cichlidae)

Copper (Cu) is one of the most commonly reported metal pollutants in African water bodies, but there are few studies on African freshwater fish species of copper accumulation and copper toxicity. Adult O. mossambicus were exposed to 0mg l^?1 (control) and 0.75mg l^?1 Cu for 96h and 0 (control), 0.11, 0.29 and 0.47mg l^?1 copper for 64 days. Samples of liver and gills were collected after 96h, and after 1, 32 and 64 days, respectively. There were significant differences in the mean Cu accumulation values in the liver and gills between the control and the Cu-exposed fish after the 96-h exposure. In fish exposed to 0.11 and 0.29mg l^?1 Cu for 64 days there was an increase in copper level in the tissues. In fish exposed to 0.47mg l^?1 Cu the concentration in the gill and liver tissue did not increase between Day 1 and Day 32. At this time, Cu accumulation in the liver was higher than for fish exposed to 0.11 and 0.29mg l^?1 Cu for 64 days. Exposure to approximately 0.47mg l^?1 Cu for more than 32 days induced mortality.     

Effects of Benzo(a)pyrene on Seabass (Dicentrarchus labrax L.): Biomarkers,Growth and Behavior

The main purpose of this study was to investigate the effects of benzo(a)pyrene (BaP) on seabass (Dicentrarchus labrax) juveniles using parameters at different levels of biological organization. Liver antioxidant status, BaP biotransformation and accumulation, growth, and behavior were determined in juveniles after 28 d exposure to BaP (1–16 μ g/l). Liver ethoxyresorufin O-deethylase increased in seabass exposed to 1–8 μ g/l of BaP. Liver glutathione S-transferases and catalase activities were significantly increased at 4 and 8 μ g/l, but a slight decrease was observed at the highest concentrations tested. Bile BaP metabolites were significantly different from the control group at 1 and 16 μ g/l BaP. Liver BaP metabolites and lipid peroxidation significantly increased at 8 and 16 μ g/l BaP. These results suggest that BaP metabolites' accumulation induces oxidative damage in seabass liver. Body weight and length increase were significantly reduced in fish exposed to BaP, with LOECs of 16 and 4 μ g/l, respectively. Food intake and swimming velocity were significantly decreased after exposure to BaP, with LOEC values of 16 and 8 μ g/l, respectively. Results suggest that at concentrations of BaP equal or higher than 8 μ g/l, the detoxification capacity decreases, an accumulation of liver BaP metabolites occurs causing lipid peroxidation, affecting growth and swimming capability of fish.     

Modulatory effect of copper on nonenzymatic antioxidants in freshwater fish Channa punctatus (Bloch.)

Effect of the low level of copper exposure on nonenzymatic antioxidants was studied in a freshwater fish Channa punctatus (Bloch.). Fish were exposed to cupric chloride at the concentration of 10 ppb for 4 wk (28 d) in a static culture condition. Copper significantly (p < 0.001) increased the serum ceruloplasmin level and total iron-binding capacity. A significant (p < 0.05) increase in reduced glutathione level was recorded in all of the tissues. With regard to nonprotein thiols, copper decreased their level in the liver, but increased it in the gill. The protein-bound thiols remained unaltered except for an increase in the liver. Metallothionein (MT) induction was observed in liver only. Copper exposure had no significant effect on the ascorbic acid level and induced no lipid peroxidation over control values. It is suggested that by modulating the ceruloplasmin level, copper indirectly protects the fish, as it facilitates conversion of pro-oxidant iron to nonoxidant iron. It also induces an array of antioxidants that may be beneficial to fish in the case of oxidative stress resulting from chemical pollutants.     

Impact of radiofrequency radiation on DNA damage and antioxidants in peripheral blood lymphocytes of humans residing in the vicinity of mobile phone base stations

Radiofrequency radiations (RFRs) emitted by mobile phone base stations have raised concerns on its adverse impact on humans residing in the vicinity of mobile phone base stations. Therefore, the present study was envisaged to evaluate the effect of RFR on the DNA damage and antioxidant status in cultured human peripheral blood lymphocytes (HPBLs) of individuals residing in the vicinity of mobile phone base stations and comparing it with healthy controls. The study groups matched for various demographic data including age, gender, dietary pattern, smoking habit, alcohol consumption, duration of mobile phone use and average daily mobile phone use. The RF power density of the exposed individuals was significantly higher (p < 0.0001) when compared to the control group. The HPBLs were cultured and the DNA damage was assessed by cytokinesis blocked micronucleus (MN) assay in the binucleate lymphocytes. The analyses of data from the exposed group (n = 40), residing within a perimeter of 80 m of mobile base stations, showed significantly (p < 0.0001) higher frequency of micronuclei when compared to the control group, residing 300 m away from the mobile base station/s. The analysis of various antioxidants in the plasma of exposed individuals revealed a significant attrition in glutathione (GSH) concentration (p < 0.01), activities of catalase (CAT) (p < 0.001) and superoxide dismutase (SOD) (p < 0.001) and rise in lipid peroxidation (LOO) when compared to controls. Multiple linear regression analyses revealed a significant association among reduced GSH concentration (p < 0.05), CAT (p < 0.001) and SOD (p < 0.001) activities and elevated MN frequency (p < 0.001) and LOO (p < 0.001) with increasing RF power density.     

Relative contribution of exotoxin and micropredation to icthyotoxicity of two strains of Pfiesteria shumwayae (Dinophyceae)

The mechanism by which Pfiesteria shumwayae (Glasgow and Burkholder) kills fish is controversial. Several studies have implicated a Pfiesteria-associated exotoxin in fish mortality while other studies indicate that physical attack of dinoflagellates on fish (micropredation) and not exotoxin is responsible. We examined the ichthyotoxicity of two strains of P. shumwayae (CAAE 101272 and CCMP 2089) in a bioassay system that exposed test fish to the dinoflagellates both with and without direct contact in the same aquarium at the same time. Dinoflagellate-free supernatants from both strains were also tested for toxicity. The results showed that direct contact between P. shumwayae and fish significantly enhanced fish mortality with both strains (P < 0.001). About 87.5% and 100% of fish died when exposed directly to CAAE 101272 and CCMP 2089, respectively. When protected from direct contact with Pfiesteria cells, 19% of the fish exposed to CAAE 101272 and 6% of those exposed to CCMP 2089 died. No deaths were observed in controls. Supernatant killed fish when obtained from cultures of CAAE 101272 but not when obtained from CCMP 2089.Analysis of variance showed that, for both strains, fish mortality in Pfiesteria-inoculated bioassays was significantly higher than control bioassays both with and without direct contact (P < 0.001). Differences between strains were not significant (P = 0.3). These results indicate that both strains are associated with exotoxin production. However, the dominant and most consistent mechanism of fish mortality observed in this study required physical contact between fish and Pfiesteria cells.