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Exposure to dioxins and dibenzofurans through the consumption of fish.

BACKGROUND: In some regions, including the Baltic Sea, fatty fish such as salmon and herring contain high levels of polychlorinated dibenzodioxins and dibenzofurans. We investigated human exposure to these potentially toxic substances in relation to the consumption of fish from the Baltic Sea. METHODS: Plasma levels of 10 different dibenzofurans and 7 dioxins were analyzed in three groups of Swedish men: one group with a high intake of fish (fish eaten almost daily; n = 11), one with a moderate intake of fish (about once per week; n = 9), and one with no consumption of fish (usually because of allergy; n = 9). RESULTS: Plasma levels of several of the compounds we measured were higher in the men with a high intake of fish than in those who consumed moderate amounts, and the levels were higher in those who ate moderate amounts of fish than in those who ate none. The median amounts of the most toxic dioxin (2,3,7,8-tetrachlorodibenzo-p-dioxin) were 8.0 pg per gram of plasma lipid (range, 2.0 to 13) in the high-intake group, 2.6 pg per gram (range, 1.2 to 4.2) in the moderate-intake group, and 1.8 pg per gram (range, 1.0 to 2.5) in the nonconsumers (P = 0.001 and 0.02, respectively). There were consistent and statistically significant associations between the reported amount of fish eaten and the plasma levels of several of the dibenzofurans and dioxins. CONCLUSIONS: Contaminated fish such as those from the Baltic Sea are an important source of exposure to polychlorinated dibenzofurans and dibenzodioxins in persons who eat fish regularly. However, the clinical consequences of such exposure remain uncertain.

Adult↗

Ecotoxicological assessment of sediment, suspended matter and water samples in the upper Danube River. A pilot study in search for the causes for the decline of fish catches.

GOALS, SCOPE AND BACKGROUND: Fish populations, especially those of the grayling (Thymallus thymallus), have declined over the last two decades in the upper Danube River between Sigmaringen and Ulm, despite intensive and continuous stocking and improvement of water quality since the 1970s. Similar problems have been reported for other rivers, e.g. in Switzerland, Great Britain, the United States and Canada. In order to assess if ecotoxicological effects might be related to the decline in fish catch at the upper Danube River, sediment, suspended matter and waste water samples from sewage treatment plants were collected at selected locations and analyzed in a bioanalytical approach using a battery of bioassays. The results of this pilot study will be used to decide if a comprehensive weight-of-evidence study is needed. METHODS: Freeze-dried sediments and suspended particulate matters were extracted with acetone in a Soxhlet apparatus. Organic pollutants from sewage water were concentrated using XAD-resins. In order to investigate the ecotoxicological burden, the following bioassays were used: (1) neutral red assay with RTL-W1 cells (cytotoxicity), (2) comet assay with RTL-W1 cells (genotoxicity), (3) Arthrobacter globiformis dehydrogenase assay (toxicity to bacteria), (4) yeast estrogen screen assay (endocrine disruption), (5) fish egg assay with the zebrafish (Danio rerio; embryo toxicity) and (6) Ames test with TA98 (mutagenicity). RESULTS AND DISCUSSION: The results of the in vitro tests elucidated a considerable genotoxic, cytotoxic, mutagenic, bacteriotoxic, embryotoxic and estrogenic burden in the upper Danube River, although with a very inhomogeneous distribution of effects. The samples taken from Riedlingen, for example, induced low embryo toxicity, but the second highest 17beta-estradiol equivalent concentration (1.8 ng/L). Using the fish egg assay with native sediments, a broad range of embryotoxic effects could be elucidated, with clear-cut dose-response relationships for the embryotoxic effects of contaminated sediments. With native sediments, embryotoxicity was clearly higher than with corresponding pore waters, thus corroborating the view that--at least for fish eggs--the bioavailability of particle-bound lipophilic substances in native sediments is higher than generally assumed. The effect observed most frequently in the fish egg assay was a developmental delay. A comparison of our own results with locations along the rivers Rhine and Neckar demonstrated similar or even higher ranges of ecotoxicological burdens in the Danube River. CONCLUSIONS: The complex pattern of ecotoxicological effects caused by environmental samples from the Danube River, when assessed in an in vitro biotest battery using both acute and more specific endpoints, showed that integration of different endpoints is essential for appropriate hazard assessment. Overall, the ecotoxicological hazard potential shown has indeed to be considered as one potential reason for the decline in fish catches at the upper Danube River. However, based on the results of this pilot study, it is not possible to elucidate that chemically induced alterations are responsible for the fish decline. RECOMMENDATIONS AND PERSPECTIVES: In order to confirm the ecological relevance of the in vitro results for the situation in the field and especially for the decline of the grayling and other fishes, further integrated investigations are required. For linking the weight of evidence obtained by in vitro assays and fish population investigations, the application of additional, more specific biomarkers (e.g. vitellogenin induction, EROD and micronucleus assay) has been initiated in fish taken from the field as well as in situ investigations.

Animals↗

Demonstration of toxicity to fish and to mammalian cells by Pfiesteria species: comparison of assay methods and strains.

Toxicity and its detection in the dinoflagellate fish predators Pfiesteria piscicida and Pfiesteria shumwayae depend on the strain and the use of reliable assays. Two assays, standardized fish bioassays (SFBs) with juvenile fish and fish microassays (FMAs) with larval fish, were compared for their utility to detect toxic Pfiesteria. The comparison included strains with confirmed toxicity, negative controls (noninducible Pfiesteria strains and a related nontoxic cryptoperidiniopsoid dinoflagellate), and P. shumwayae strain CCMP2089, which previously had been reported as nontoxic. SFBs, standardized by using toxic Pfiesteria (coupled with tests confirming Pfiesteria toxin) and conditions conducive to toxicity expression, reliably detected actively toxic Pfiesteria, but FMAs did not. Pfiesteria toxin was found in fish- and algae-fed clonal Pfiesteria cultures, including CCMP2089, but not in controls. In contrast, noninducible Pfiesteria and cryptoperidiniopsoids caused no juvenile fish mortality in SFBs even at high densities, and low larval fish mortality by physical attack in FMAs. Filtrate from toxic strains of Pfiesteria spp. in bacteria-free media was cytotoxic. Toxicity was enhanced by bacteria and other prey, especially live fish. Purified Pfiesteria toxin extract adversely affected mammalian cells as well as fish, and it caused fish death at environmentally relevant cell densities. These data show the importance of testing multiple strains when assessing the potential for toxicity at the genus or species level, using appropriate culturing techniques and assays.

Animals↗

Peroxisome proliferator-activated receptor alpha is not rate-limiting for the lipoprotein-lowering action of fish oil.

Similar to fibrate hypolipidemic drugs, long chain polyunsaturated fatty acids contained in fish oil are activators of peroxisome proliferator-activated receptor alpha (PPARalpha). The goal of this study was to assess the contribution of PPARalpha in mediating the effect of fish oil on plasma lipid, lipoprotein, and apolipoprotein levels. To this end, PPARalpha-deficient mice and wild-type littermates were fed isocaloric fish oil or coconut oil diets, the content of which varied reciprocally between 0, 3, 7, and 10% for 1 week. In both wild-type and PPARalpha-deficient mice, fish oil feeding was associated with a dose-dependent decrease in triglycerides, cholesterol, and phospholipids associated with lower levels of very low density lipoprotein (VLDL) triglycerides and high density lipoprotein (HDL) cholesterol. The lowering of triglycerides and VLDL triglycerides was associated with a significant decrease of plasma apoC-III in both genotypes. Fish oil treatment did not influence hepatic apoC-III mRNA levels in either genotype indicating that apoC-III is not under transcriptional control by fish oil. The lowering of HDL cholesterol observed in both genotypes was associated with reduced plasma apoA-II without changes in liver apoA-II mRNA levels. In contrast, plasma apoA-I and liver apoA-I mRNA levels were decreased in wild-type but not in PPARalpha-deficient mice after fish oil feeding indicating that PPARalpha contributes to the effect of fish oil on apoA-I gene expression. In conclusion, PPARalpha is not rate-limiting for fish oil to exert its triglyceride- and HDL-lowering action. Furthermore, PPARalpha mediates, at least partly, the decrease of apoA-I after fish oil treatment, whereas apoC-III and apoA-II levels are affected in a PPARalpha-independent manner. Altogether, these results show major molecular differences in action between fibrates and fish oil providing a molecular rationale for combination treatment with these compounds.

Acyl-CoA Oxidase↗

Fishing and risk along the Savannah River: possible intervention.

Fishing is often perceived as an enjoyable activity, and eating fish is viewed as safe and healthful. However, with recent increases in consumption advisories because of contamination, the public is faced with whether to eat fish or not. In this article I examine the knowledge base of people fishing along the Savannah River, where South Carolina has issued consumption advisories because of mercury and radionuclides. Over 250 people fishing from the Augusta lock and dam to south of the Department of Energy's Savannah River Site (SRS) were interviewed from early April until late November 1997. Overall 82% of the fishermen thought the fish were safe to eat, even though 62% had heard some warnings about eating the fish. There were significant differences in whether people thought the fish were safe to eat as a function of income, age, education, and whether they were employed at the Savannah River Site. Significantly more fishermen thought the fish were safe who made more than $20,000/year, were over 34 yr of age, worked at SRS, and had no college or technical training, compared to others. Significantly fewer blacks had heard of consumption advisories than whites, fewer low-income people had heard, and fewer people who had not worked at SRS had heard, compared to others. Most people heard about the advisories from television, newspapers, and other people, although more blacks than whites heard about advisories from the radio. There were also significant ethnic differences in distance traveled, and in whether specific fish were frozen for later consumption. These data can be used to design an information program to target the people who may be most at risk from eating fish obtained from the Savannah River.

Adolescent↗

Mercury levels in hair from people eating large quantities of Swedish freshwater fish.

Mercury concentrations in hair were related to fish-eating habits in a group of 50 people reported to have a high consumption of freshwater fish. Mercury levels in hair ranged from 0.3 to 10.8 mg/kg with a mean +/- SD of 3.2 +/- 2.3 mg/kg. The average mercury level in hair from men was significantly higher than that in hair from women (3.8 +/- 2.6 mg/kg versus 2.4 +/- 1.8 mg/kg. Seven individuals (14%) had levels above 6 mg/kg. In people with equal fish consumption, significantly higher mercury levels were found in the hair of those eating fish from lakes Mälaren and Vättern than in those eating fish from Lake Hjälmaren. It has been reported that fish from the latter lake contains approximately 0.2 mg/kg mercury, whereas fish from the other two lakes contains approximately 0.4 mg/kg. The mean mercury level in hair was higher in the group eating freshwater fish more than three times a week (greater than or equal to 500 g fish flesh/week) than in the group eating less, although the difference was of borderline significance. Within couples (n = 16) eating equal numbers of fish meals per week, the men had significantly higher levels (mean 3.7 mg/kg) than the women (mean 2.5 mg/kg). The results from the present study show that people with a high consumption of Swedish freshwater fish have elevated levels of mercury in their hair, when compared with previously reported levels in the hair of Swedish pregnant women.

Adult↗

An event-by-event probabilistic methodology for assessing the health risks of persistent chemicals in fish: a case study at the Palos Verdes Shelf.

A human health risk assessment of recreational anglers who consume fish from the Palos Verdes Shelf was conducted. The uptake of DDT, DDE, and DDD (collectively total DDT or tDDT) and polychlorinated biphenyls (PCBs) due to fish ingestion was characterized using Monte Carlo techniques. This analysis relied upon 176 probability density functions developed from over 300,000 individual pieces of information to represent 17 different exposure factors that influence the human uptake of persistent organic chemicals in fish. The carcinogenic and noncarcinogenic risks were estimated using a microexposure event modeling approach that estimates exposure on an event-by-event basis. This evaluation relied upon several large studies that provided site-specific data on angler behavior and concentratioins of chemicals in 13 fish species. Our results indicate that the median theoretical increased lifetime cancer risk associated with estimated exposure to tDDT and PCBs was 5 x 10(-8) for anglers who fish on commercial passenger fishing vessels (CPFVs) and who catch and eat fish from the Palos Verdes Shelf. The mean risk for these anglers was 2 x 10(-7), and the 95th percentile risk was 8 x 10(-7). At the 9.5th percentile, the hazard quotients for anglers were less than 1, indicating that noncancer effects are unlikely. These results are in contrast with prior risk assessments of this site that suggested that consumption of white croaker alone posed a cancer risk of 2 x 10(-3) and a hazard quotient of 32. Our results were validated by their agreement with several independent local studies regarding fishing and consumption practices. This assessment indicates that the levels of tDDT and PCB in fish at the Palos Verdes Shelf do not pose a significant risk to human health among recreational anglers. Based on the size of the local angler population, no cases of cancer would be expected to result from eating Palos Verdes Shelf fish. The methodology used here should be applicable to characterizing the risks to those who ingest fish from the waterways of most industrialized nations.

Animals↗

Dietary fish oil decreases low-density-lipoprotein clearance in nonhuman primates.

To assess whether fish oil-induced alterations in low-density-lipoprotein (LDL) composition have distinct and important effects on LDL metabolism, we evaluated LDL kinetic behavior in cynomolgus macaques fed an atherogenic diet supplemented with either fish oil (1.6 g n-3 fatty acids; n = 10) or olive oil (n = 9) for > or = 6 mo. LDL from monkeys supplemented with fish oil or olive oil was isolated, labeled with either 125I or 131I, and simultaneously reinjected so that each monkey received its own (autologous injection) and donor (homologous injection) LDL. For LDL injected autologously (monkeys that received their own LDL), the LDL fractional clearance rate (FCR) was reduced in fish oil-supplemented monkeys compared with the olive oil-supplemented controls (0.42 +/- 0.03 compared with 0.56 +/- 0.05 pools/d, P = 0.04). The cholesteryl ester content of fish oil LDL increased compared with olive oil LDL (43 +/- 2% and 36 +/- 3%, respectively, P = 0.03), and the LDL cholesteryl ester content was strongly correlated with autologous LDL clearance (r = -0.76, P = 0.0001). Compared with olive oil LDL, fish oil LDL had a reduced dissociation constant (KD) for binding to the LDL receptor in vitro (KD for fish oil LDL compared with olive oil LDL: 13.9 +/- 1.8 and 7.4 +/- 1.0 mg LDL protein/L, P = 0.03). When both fish oil LDL and olive oil LDL were simultaneously injected into fish oil-supplemented monkeys, the FCR of fish oil LDL was decreased compared with olive oil LDL (0.42 +/- 0.03 and 0.52 +/- 0.04 pools/d, P = 0.006). These data suggest that dietary supplementation with fish oil decreases LDL clearance, and that this effect is mediated, at least in part, by altering LDL structure and reducing the affinity of LDL for its receptor.

Animals↗

Fish consumption and coronary heart disease mortality in Finland, Italy, and The Netherlands.

Fish consumption seems to protect against death from coronary heart disease (CHD). If this association is due to n-3 polyunsaturated fatty acids, especially fatty fish may be responsible for this protective effect. The association between total, lean, and fatty fish consumption and the risk of CHD mortality was examined in 1,088 Finnish, 1,097 Italian, and 553 Dutch men participants in the Seven Countries Study who were aged 50-69 years and free of CHD around 1970. After 20 years of follow-up, 242 (22.2%) men in Finland, 116 (10.6%) men in Italy, and 105 (19.0%) men in the Netherlands had died of CHD. Cox proportional hazards analysis showed no association between total fish consumption and CHD mortality. After adjustments were made for age, body mass index, smoking, energy intake, and relevant dietary variables, the pooled relative risk for the highest quartile of total fish compared with no fish consumption in the three countries was 1.08 (95% confidence interval: 0.76, 1.53). Lean fish consumption also was not associated with CHD mortality in any country. Fatty fish compared with non-fatty-fish consumption was associated with lower CHD mortality; the adjusted, pooled relative risk for fatty fish consumers was 0.66 (95% confidence interval: 0.49, 0.90). These data suggest that especially fatty fish is protective against CHD mortality.

Adult↗

Development of a single-meal fish consumption advisory for methyl mercury.

Methyl mercury (meHg) contamination of fish is the leading cause of fish consumption advisories in the United States. These advisories have focused upon repeated or chronic exposure, whereas risks during pregnancy may also exist from a single-meal exposure if the fish tissue concentration is high enough. In this study, acute exposure to meHg from a single fish meal was analyzed by using the one-compartment meHg biokinetic model to predict maternal hair concentrations. These concentrations were evaluated against the mercury hair concentration corresponding to the U.S. Environmental Protection Agency's reference dose (RfD), which is intended to protect against neurodevelopmental effects. The one-compartment model was validated against blood concentrations from three datasets in which human subjects ingested meHg in fish, either as a single meal or multiple meals. Model simulations of the single-meal scenario at different fish meHg concentrations found that concentrations of 2.0 ppm or higher can be associated with maternal hair concentrations elevated above the RfD level for days to weeks during gestation. A single-meal fish concentration cutoff of > or = 2.0 ppm is an important consideration, especially because this single high exposure event might be in addition to a baseline meHg body burden from other types of fish consumption. This type of single-meal advisory requires that fish sampling programs provide data for individual rather than composited fish, and take into account seasonal differences that may exist in fish concentrations.

Adult↗

Radiocesium in fish from the Savannah River and Steel Creek: potential food chain exposure to the public.

This study examined radiocesium (137Cs) levels in fish from the vicinity of the Department of Energy's Savannah River Site (SRS), a former nuclear weapons production facility in South Carolina. Fish from the Savannah River were sampled above (upstream), along, and below (downstream) the SRS, and from Steel Creek, a tributary that runs through the SRS. There was some off-site contamination of 137Cs in the Savannah River watershed due to low-level releases from past nuclear production on the SRS. The null hypotheses tested were that there would be no differences in 137Cs levels as a function of location along the river, and between species collected from the river and from Steel Creek on the SRS. For six of eight species of fish collected from the Savannah River, there were no differences in 137Cs levels in muscle from fish collected above, along, or below the SRS; exceptions were bowfin and shellcracker. Fish collected from Steel Creek had significantly higher levels (by about an order of magnitude) of 137Cs in muscle tissue than fish collected in the Savannah River. However, no fish from either Steel Creek or the Savannah River had 137Cs levels above the European Economic Community limit for fresh meat of 0.6 Bq/g. Lifetime cancer risk was calculated using the cancer slope factor of 3.2 x 10(-11)/pCi, and various fish consumption scenarios reflecting actual data from Savannah River fishermen. Using mean 137Cs concentrations and median fish consumption for 70 years for Black males-the group with the highest consumption-the excess lifetime risk associated with the eight species of fish in the Savannah River ranged from 9.0 x 10(-7) to 1.0 x 10(-5). The same calculation for fish from Steel Creek gave risk estimates from 1.4 to 8.0 x 10(-5). The 95% level for consumption by Blacks, however, was about 70 kg/year. Black fishermen consuming that amount of bass from Steel Creek would sustain a lifetime risk of 3.1 x 10(-4), whereas the same consumption of Savannah River bass would yield a risk estimate of 1.5 x 10(-5).

Animals↗

Evaluating risk communication about fish consumption advisories: efficacy of a brochure versus a classroom lesson in Spanish and English.

Presentation format can influence the way target audiences understand risk-related information. Brochures or fish fact sheets are the methods traditionally used by state agencies to inform the public about fish consumption advisories and the risks from consuming fish. This study examines the efficacy of presenting information about the risks from consuming contaminated fish and shellfish in two different formats: a brochure and classroom presentation. The two instruments were developed and tested in Spanish and English, reflecting the local ethnic composition in the Newark Bay Complex. The instruments were tested on women of child-bearing age at the Women, Infants, and Children Center in Elizabeth, New Jersey. Detailed diagrams were used in both presentations, including contaminated fish species, fish preparation methods, and food chain bioaccumulation and transmission to the fetus. There were few language-related differences in the efficacy of the classroom lesson, and the main ideas were understood by both groups. Where there were significant differences in understanding about the risks from consuming fish or crabs from the contaminated waters of Newark Bay, in all cases the women exposed to the classroom lesson had a better understanding than those who read the brochure. Ninety-six percent of the women who heard the lesson understood that it was unsafe to eat fish from the port, compared to 72% of those reading the brochure. Both formats succeeded in imparting information to most women about the area under advisories, the fish species under advisories, and transmission of toxins to the fetus. Information on fish preparation was recalled less clearly, partly because women were asked to relate methods to reduce the risk from consuming fish from 11 presented, and most recalled only two or three of the list. The advantages and disadvantages of conducting short classes to women of child-bearing age are discussed.

Adult↗

Adaptation of rainbow fish to lake and stream habitats.

Fish occupy a range of hydrological habitats that exert different demands on locomotor performance. We examined replicate natural populations of the rainbow fishes Melanotaenia eachamensis and M. duboulayi to determine if colonization of low-velocity (lake) habitats by fish from high-velocity (stream) habitats resulted in adaptation of locomotor morphology and performance. Relative to stream conspecifics, lake fish had more posteriorly positioned first dorsal and pelvic fins, and shorter second dorsal fin bases. Habitat dimorphism observed between wild-caught fish was determined to be heritable as it was retained in M. eachamensis offspring raised in a common garden. Repeated evolution of the same heritable phenotype in independently derived populations indicated body shape divergence was a consequence of natural selection. Morphological divergence between hydrological habitats did not support a priori expectations of deeper bodies and caudal peduncles in lake fish. However, observed divergence in fin positioning was consistent with a family-wide association between habitat and morphology, and with empirical studies on other fish species. As predicted, decreased demand for sustained swimming in lakes resulted in a reduction in caudal red muscle area of lake fish relative to their stream counterparts. Melanotaenia duboulayi lake fish also had slower sustained swimming speeds (Ucrit) than stream conspecifics. In M. eachamensis, habitat affected Ucrit of males and females differently. Specifically, females exhibited the pattern observed in M. duboulayi (lake fish had faster Ucrit than stream fish), but the opposite association was observed in males (stream males had slower Ucrit than lake males). Stream M. eachamensis also exhibited a reversed pattern of sexual dimorphism in Ucrit (males slower than females) relative to all other groups (males faster than females). We suggest that M. eachamensis males from streams responded to factors other than water velocity. Although replication of muscle and Ucrit phenotypes across same habitat populations within and/or among species was suggestive of adaptation, the common garden experiment did not confirm a genetic basis to these associations. Kinematic studies should consider the effect of the position and base length of dorsal fins.

Adaptation, Physiological↗

Blood pressure and vascular reactivity changes in spontaneously hypertensive rats fed fish oil.

1. To examine possible mechanisms of antihypertensive effects of feeding fish oil rich in n-3 fatty acids, we have studied vascular reactivity of aortic rings and perfused mesenteric resistance vessels of spontaneously hypertensive rats (SHR) given such a diet. 2. In two experiments, rats were fed a semi-synthetic diet containing either 'fish oil' (10 and 20% by weight) or hydrogenated coconut oil (control) (10 and 20%) for 4 weeks. 3. Blood pressure rose significantly less in the fish oil group than in controls in both experiments. 4. Aortic rings from control rats showed endothelium-dependent relaxations to low concentrations of acetylcholine (ACh) but relaxed less at higher concentrations. In contrast, rings from the fish oil group had relaxations which increased through the range of concentrations used. Indomethacin (10 microM) also increased the relaxation responses seen in rings from control rats, suggesting that fish oil inhibits a contractile cyclo-oxygenase product. This contractile substance may be thromboxane A2 (TxA2) or its endoperoxide precursor, prostaglandin H2 (PGH2) as aortic incubates and serum levels of TxB2 (the stable product of TxA2) were greatly reduced in fish oil-fed rats, and the decrease of relaxant responses to high concentrations of ACh were also blocked by a TxA2/PGH2 receptor blocker (SQ 29548). 5. In contrast to aortic rings, perfused preconstricted mesenteric resistance vessels of control rats relaxed to ACh in a similar fashion to tissues from fish oil-fed rats. However, in this preparation, fish oil feeding enhanced relaxations to sodium nitroprusside (SNP) and contractile responses to noradrenaline were less than controls. After removal of endothelium with 0.05% saponin, contractile responses to noradrenaline increased in both groups but responses from fish oil-treated rats were still attenuated. This suggests that fish oil feeding alters reactivity of mesenteric resistance vessels at the level of the smooth muscle. 6. The results indicate that fish oil feeding may reduce blood pressure by decreasing vascular smooth muscle reactivity to noradrenaline in resistance vessels. The effect may be enhanced by inhibition of an endothelium-derived cyclo-oxygenase product, such as TxA2 or PGH2 in conduit vessels.

6-Ketoprostaglandin F1 alpha↗

N-3 fatty acids from fish oil. Effects on plasma lipoproteins and hypertriglyceridemic patients.

In the experimental studies reported in this review, dietary n-3 fatty acids from fish and fish oil had profound hypolipidemic effects in normal subjects and in hypertriglyceridemic patients with combined hyperlipidemia (type II-b) and types IV and V hyperlipidemia. In these carefully controlled metabolic experiments, dramatic reductions occurred in plasma triglycerides and to a lesser extent in plasma total cholesterol. Reductions in VLDL, chylomicrons, remnants, LDL, apo B, and apo E were also noted. HDL changes varied from subject to subject. These plasma lipoprotein changes occurred in subjects with non-insulin-dependent diabetes mellitus as well, without deterioration of diabetic control. Similar results are reported in two other papers in this volume. Fish oil did not cause deterioration of diabetic control. Whereas the mechanism of the hypolipidemic action of the n-6 rich vegetable oils containing linoleic acid such as corn or safflower oil still remains obscure, the mechanism of the hypolipidemic action of the n-3 fatty acids in fish oil is well documented. The synthesis of triglyceride and VLDL in the liver is greatly reduced by n-3 fatty acids. At the same time, the turnover of VLDL in plasma is shortened. In another study, LDL production was decreased. Combined with other dietary manipulations, such as a reduction in saturated fat and dietary cholesterol, the use of n-3 fatty acids to treat hyperlipidemia, especially hypertriglyceridemia, appears to have a well-supported rationale. Fish oil combined with a low cholesterol, low saturated fat diet has been shown to produce complementary effects. Total plasma cholesterol and LDL cholesterol were lowered by the low cholesterol, low saturated fat diet, whereas plasma triglyceride and VLDL were decreased by the fish oil. In most situations, the use of fish oil supplements should be regarded as pharmacologic therapy, particularly effective in severe hypertriglyceridemic states (e.g., chylomicronemia). However, a lifelong diet rich in fish may be protective against atherosclerosis as well. Further studies are required to delineate exact doses and precise indications for the use of fish oil in different types of hyperlipidemias and to differentiate the effects, if any, of the two major n-3 fatty acids in fish oil, EPA and DHA. The hypolipidemic effects of n-3 fatty acids coupled with their known antithrombotic actions (secondary to changes in prostaglandin secretion, platelet function, inhibition of growth factors, and enhancement of endothelial-derived relaxation factor) appear to have an important potential role in the control of coronary heart disease and other atherosclerotic disorders. Moreover, fish oil may prevent the "chylomicronemia" syndrome of type V hyperlipidemia.

Dietary Fats, Unsaturated↗

DNA vaccines encoding viral glycoproteins induce nonspecific immunity and Mx protein synthesis in fish.

Protective immunity by vaccination with plasmid DNA encoding a viral glycoprotein (G) has long been assumed to result from the induction of a specific immune response. We report here that the initial protection may be due to the induction of alpha/beta interferon, with long-term protection due to a specific response to the encoded viral G. DNA vaccines encoding the Gs of three serologically unrelated fish rhabdoviruses were used to vaccinate rainbow trout against a lethal challenge with infectious hematopoietic necrosis virus (IHNV). All three vaccines, each encoding the G gene of either IHNV (IHNV-G), snakehead rhabdovirus (SHRV) (SHRV-G), or spring viremia of carp virus (SVCV) (SVCV-G), elicited protective immunity against IHNV. Vaccinated fish were challenged at 30 or 70 days postvaccination with lethal doses of IHNV. At 30 days postvaccination, only 5% of fish that had received any of the G vaccines died, whereas more than 50% of the control fish succumbed to virus challenge. When fish were vaccinated and challenged at 70 days postvaccination, only 12% of the IHNV-G-vaccinated fish died compared to 68% for the SHRV-G- and 76% for the SVCV-G-vaccinated fish. Assays for trout Mx protein, an indicator of alpha/beta interferon induction, showed that only fish vaccinated with a G-containing plasmid produced high levels of Mx protein in the kidneys and liver. Interestingly, at day 7 after virus challenge, all of the fish vaccinated with the IHNV-G plasmid were negative for Mx, but the SHRV-G- and SVCV-G-vaccinated fish still showed detectable levels of Mx. These results suggest that DNA vaccines in fish induce an early, nonspecific antiviral protection mediated by an alpha/beta interferon and, later, a specific immune response.

Amino Acid Sequence↗

Protein and amino acid requirements of fishes.

Tentative qualitative and quantitative amino acid requirements have been reported for the major species of fish reared for market or as replacement stocks for natural waters. Most work has concentrated upon juvenile fish or upon rapidly growing young market fish; these have high protein dietary requirements (30-50%) that are in direct contrast to the homothermic terrestial animals. Net protein utilization from the diet is similar or slightly better than that found in avian species, but energy needs are much lower in fish and as a result the body protein deposition in fish is larger (about 5 g protein/MJ for the chick versus about 10 g protein/MJ for young fish). Qualitative amino acid requirements appear identical for all fish species examined; arginine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan, and valine are all required for normal growth and metabolism. Quantitative requirements differ only slightly among species that have been tested. Salmon have higher arginine requirements than other fish examined. Catfish appear to have a lower requirement for histidine and threonine, and the Japanese eel seems to need more tryptophan in the diet. However, when the quantitative requirements for indispensable amino acids are expressed as a percentage of the protein fed, then a remarkable harmony appears between values needed for maximal growth for most species examined. A review of Tables 3 and 4 will disclose the paucity of information available considering the large number of fish species reared commercially over the world. Most commercial diet formulations have relied upon the work done on salmon, catfish, and carp, and their amino acid and protein requirement values have been used. Remarkably, these diets have produced other species of fish economically. Sparing effects of one amino acid on another have only been studied with cystine-methionine and tyrosine-phenylalanine. Arginine and analogues of methionine have been used as good nitrogen sources for salmon. Isoleucine-leucine ratios have been measured and experiments indicate some growth inhibition when the isoleucine-leucine ratio was greater than 2/1. Valine at abnormally high levels also inhibited growth. Much more work needs to be done on the effects of subtle differences in amino acid ratios in the diet, and major emphasis should be placed on the important role of the dispensable amino acids in fish nutrition.(ABSTRACT TRUNCATED AT 400 WORDS)

Aging↗

Effect of dietary fish oil on coronary artery and aortic atherosclerosis in African green monkeys.

Studies were carried out for 2.5 to 3 years in adult male African green monkeys (grivet subspecies) fed diets containing 22% of calories as lard or fish oil with 40% of calories as fat and 0.75 mg cholesterol/Kcal to determine if isocaloric substitution of menhaden fish oil for lard affects coronary artery atherosclerosis. The average total plasma cholesterol concentrations during the experimental period were significantly lower for the fish-oil group (231 +/- 37 mg/dl) compared to the lard group (360 +/- 44 mg/dl), but this difference did not become apparent until after 5 months of experimental diet consumption. High density lipoprotein cholesterol concentrations were 30% lower (p less than 0.01) for the fish-oil group also (57 +/- 5 vs. 82 +/- 6 mg/dl). Plasma triglyceride concentrations were low for both groups, but after about 5 months of diet consumption, they were higher for the animals fed fish oil (25 +/- 2 mg/dl) compared to their lard-fed counterparts (15 +/- 1 mg/dl). Coronary artery intimal area (in this case a measure of early atherosclerotic lesion size) was low in all animals but was significantly less (p less than 0.03) for the fish oil vs. lard groups (0.01 +/- 0.002 vs. 0.03 +/- 0.009 mm2). More atherosclerosis was found in other arteries, and a trend was seen of less atherosclerosis in the thoracic aorta and common carotid arteries of the fish-oil group. The size of lesions in the abdominal aorta was similar between diet groups, but microscopic examination of arteries of the lard group revealed relatively more cholesterol monohydrate crystals compared to the arteries of the fish-oil group. Chemical analysis showed that there was less esterified cholesterol (1.46 +/- 0.71 vs. 3.43 +/- 0.74 mg/g, p = 0.04) and free cholesterol (3.7 +/- 2.15 vs. 7.05 +/- 1.68 mg/g, p = 0.08) in the abdominal aortas taken from the animals fed fish oil. There was a significant correlation between low density lipoprotein (LDL) cholesteryl ester (CE) fatty acid ratio (i.e., saturated + monounsaturated/polyunsaturated species) and the amount of esterified (r = 0.59) and free (r = 0.63) cholesterol in the abdominal aortas. Compared to the lard group, animals fed fish oil had significantly lower LDL CE melting temperatures (26 +/- 1 vs. 38 +/- 1 degree C) and significantly smaller LDL particles (2.68 +/- 0.10 vs. 3.25 +/- 0.38 g/mumol). Therefore, the potentially antiatherogenic effects of dietary fish oil include its ability to decrease the concentration, size, CE content, and CE melting temperature of plasma LDL.

Animals↗