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[Aspects of animal welfare and species protection in the international trade of ornamental fish and air transport to Germany].

The number of ornamental fish kept in German aquariums is rising, but the supply of offspring is not sufficient to meet the needs of the fish enthusiasts. Therefore millions of ornamental fish from foreign countries are being imported to Germany. This provokes a number of new problems regarding the protection of species and the animals' welfare during transportation. For the assessment of the transport conditions, 1000 shipments of ornamental fishes were evaluated at the Rhein-Main-Airport Frankfurt, Germany. Water samples were taken from 100 shipments and were examined for anaesthetics. The results are disturbing: 41% of the shipments have total transportation times between 31-42 h resulting in an oxygen deficit in the transport bags (5.2%) and dead ornamental fish. Also damage of the transportation containers, high fish densities as well as drastic size differences amoung the fish in one transport bag were noticeable. In 99% of the cases, formal defects could be noted. The German association of pet stores has declared that fish belonging to three special fresh water families should not be kept in an aquarium, but 1 200 of such fish were imported. The HPLC analysis of the water samples revealed an additive. The non-declared anaesthetic 2-phenoxyethanol was present in all samples taken from shipments based out of Singapore. The results emphasize that improvements are urgently necessary in the control of the air transport and trade with Ornamental fish.

Animal Welfare↗

[Study on the relationship between contents of poison in fishes and the levels of ocean pollutants in Zhoushan Fishery].

OBJECTIVE: To investigate the contents of poison in fishes in the Zhoushan Fishery and to analysis its relationship with the degree of ocean pollution. METHOD: Nine kinds of major fishes in the Zhoushan Fishery were sampled and tested. Canonical correlation was conducted to evaluate the correlation between contents of poison in fishes and the degree of ocean pollution. RESULTS: The average contents of nickel among 9 kinds of fishes exceeded normal value by 1.177 times. No significantly statistical differences were found between the levels of poison contents in nine kinds of major fishes during 1997 to 2003 in the Zhoushan Fishery (the P values are all less than 0.05), except that the levels of cadmium and hydrargyrum in some fishes (the P values are all more than 0.05). The coefficient of fishes accumulated from different poisons ranged from 101.80 to 71 210.97. The canonical correlation coefficients of the variables of the contents of plumbum, arsenic and benzene hexachloride, chlorophennothane between fishes and ocean were 0.784 and 0.808 respectively. Positive correlation between them was shown (chi(2) = 20.994, P < 0.01; chi(2) = 38.017, P < 0.01). CONCLUSION: The sanitary quality indexes of all poisons which had been tested fell in the normal ranges except for nickel. The degrees of poisons accumulated among distinct fishes varied, showing the relationship of medium positive correlation between the poison contents of fishes and the degree of the ocean pollution.

Animals↗

Assessment of pollution levels in fish and water of main basin, Lake Mariut.

Lake Mariut has suffered over the years from the untreated sewage, agricultural and industrial wastes dumped into it. Lake Mariut fish quality and quantity were adversely affected eventually being unfit for human consumption due to its poor water quality. The present study was carried out to evaluate the levels of metals in the fish caught from Lake Mariut main basin and its fitness for human consumption, and the water quality after the enforcement of the Egyptian Law for Environmental Protection (Law 4/1994). Fish and water samples were collected form Lake Mariut main basin through out the period from May to December 2000. The results showed that mercury was totally undetected in fish and water samples. The levels of Fe obtained were very low compared to previous studies. Fe and Cr contents in fish flesh were higher compared to their levels in water samples. Also Cu has shown a decrease in both fish and water samples. Zn and Cd contents in water samples have decreased compared with former studies. However, they have not changed in fish flesh. Pb content in both fish flesh and water samples has shown a great decrease compared to previous studies. The Cd and Pb mean values in fish flesh of the present study (0.81 and 0.14 mg/kg) exceeded the Egyptian Standards No. 2360/1993 value (should not exceed 0.1 mg/kg for each). The results also showed that the water quality has changed compared with previous studies. BOD has increased in north of Main Basin, east of Main Basin, South of Main Basin, and southwest of Main Basin; and deCreased in WTP effluent; and Qalaa Drain. There has been an increase in TSS, TVSS, and NO3 levels compared to pervious studies. There has been a decrease in TS, TDS, COD,hardness, and chloride levels. This decrease, especially in COD, may be attributed to the enforcement of the Egyptian Law 4/1994. The pH and alkalinity were in agreement with the values shown by previous studies except for east of Main Basin which showed higher values. The study included some recommendations aiming at improving both water quality and fish of the Main Basin.

Animals↗

[Fish resources and their conservation strategies in Hepu Dugong State Nature Reserve and its adjacent waters].

An investigation was made on the fish resources and fish habitats of Hepu Dugong State Nature Reserve and its adjacent waters from June 2003 to May 2004. The local fish markets were surveyed, and fishermen and fishmongers were interviewed to get fishery information. A total of 57 fish species belonging to 41 families and 14 orders were recorded, among which, 55 were bony fish, and 2 were elasmobranch. There were 41 fish species belonging to warm-water fish, and 37 fish species belonging to benthonic fish. The faunal characteristic was mainly of Northern Pacific and Indo-Pacific fauna. Devastating fishing and environmental pollution were the most serious threats to the local fishery resources, and some countermeasures were proposed for conservation and management of the resources.

Animals↗

Fish oil decreases hepatic cholesteryl ester secretion but not apoB secretion in African green monkeys.

Two groups of African green monkeys were fed diets containing 40% of calories as fat with half of the fat calories as either fish oil or lard. The fish oil-fed animals had lower cholesterol concentrations in blood plasma (33%) and low density lipoproteins (LDL) (34%) than did animals fed lard. Size and cholesteryl ester (CE) content of LDL, strong predictors of coronary artery atherosclerosis in monkeys, were significantly less for the fish oil-fed animals although the apoB and LDL particle concentrations in plasma were similar for both diet groups. We hypothesized that decreased hepatic CE secretion led to the smaller size and reduced CE content of LDL in the fish oil-fed animals. Hepatic CE secretion was studied using recirculating perfusion of monkey livers that were infused during perfusion with fatty acids (85% 18:1 and 15% n-3) at a rate of 0.1 mumol/min per g liver. The rate of cholesterol secretion was less (P = 0.055) for the livers of fish oil versus lard-fed animals (3.3 +/- 0.5 vs. 6.0 +/- 1.2 mg/h per 100 g, mean +/- SEM) but the rate of apoB secretion was similar for both groups (0.92 +/- 0.15 vs. 1.01 +/- 0.13 mg/h per 100 g, respectively). The hepatic triglyceride secretion rate was also less (P less than 0.05) for the fish oil-fed animals (8.3 +/- 2.5 vs. 18.3 +/- 4.4 mg/h per 100 g). Liver CE content was lower (P less than 0.006) in fish oil-fed animals (4.1 +/- 0.8 vs. 7.4 +/- 0.7 mg/g) and this was reflected in a lower (P less than 0.04) esterified to total cholesterol ratio of perfusate VLDL (0.21 +/- 0.045 vs. 0.41 +/- 0.06). The hepatic VLDL of animals fed fish oil had 40-50% lower ratios of triglyceride to protein and total cholesterol to protein. From these data we conclude that livers from monkeys fed fish oil secreted similar numbers of VLDL particles as those of lard-fed animals although the hepatic VLDL of fish oil-fed animals were smaller in size and relatively enriched in surface material and depleted of core constituents. Positive correlations between plasma LDL size and both hepatic CE content (r = 0.87) and hepatic VLDL cholesterol secretion rate (r = 0.84) were also found.(ABSTRACT TRUNCATED AT 400 WORDS)

Analysis of Variance↗

A fish oil diet reduces the severity of collagen induced arthritis after onset of the disease.

We have previously reported that compared to a corn oil diet a fish oil diet (5% by weight) fed to B10R.III mice before the induction of collagen induced arthritis markedly reduced disease severity. In this study we determine whether a fish oil diet could reduce the severity of collagen induced arthritis if begun after the arthritis was clinically apparent. Mice were initially fed either a fish oil or corn oil diet and immunized with bovine type II collagen 4 weeks later. At the onset of collagen-induced arthritis, half of the corn oil fed mice were switched to fish oil and arthritis assessed on a weekly basis. Four weeks after the diet change until killing 5 weeks later, the mice switched to fish oil developed much less severe arthritis than the corn oil fed controls. Thus the severity index of corn oil fed mice ranged between 9.4 and 7.1; the severity index of fish oil fed mice was between 6.8 and 4.3 while the mice switched to fish oil ranged between 7.2 and 5.6. Analysis of peritoneal macrophages 13 weeks after immunization showed that macrophages from fish oil fed mice incorporated eicosapentaenoic acid into phospholipids and produced less arachidonate products than corn oil fed mice. There was no difference between macrophages obtained from mice switched from corn oil to fish oil and those maintained on fish oil with respect to fatty acid composition of membrane phospholipids or prostaglandin profile. These results suggest that arthritis severity may be modulated after the onset of CIA by altering the PG profile of macrophages present at inflammatory sites.

Animals↗

Cold-stable microtubules from Antarctic fishes contain unique alpha tubulins.

The cytoplasmic microtubules of Antarctic fishes assemble from their tubulin subunits at physiological body temperatures in the range -2 to +2 degrees C. Our objective is to determine the structural features that enhance the assembly of Antarctic fish tubulins at low temperatures. Here we compare the structures of tubulin subunits from three Antarctic fishes (Notothenia gibberifrons, Notothenia coriiceps neglecta, and Chaenocephalus aceratus), from three temperate fishes (the dogfish shark Mustelus canis, the channel catfish Ictalurus punctatus, and the goosefish Lophius americanus), and from a mammal (the cow Bos taurus). When reduced, carboxymethylated, and examined by polyacrylamide gel electrophoresis, multiple alpha chains were observed in tubulins from the Antarctic fishes, the catfish, and the goosefish; dogfish and bovine alpha tubulins migrated as single components on this gel system. Prominent in the Antarctic fish tubulins was an alpha variant that migrated more rapidly than the bovine alpha chain; smaller amounts of a rapidly migrating alpha chain were also present in catfish and goosefish tubulins. The beta tubulins of the fishes, with the exception of the goosefish, resolved into major and minor variants with mobilities similar to those of beta 1 and beta 2 tubulins from bovine brain. Peptide mapping demonstrated that the alpha tubulins of Antarctic fishes were similar in structure, yet differed from the alpha chains of the dogfish and the cow (which, in turn, were similar to each other). In contrast, the beta tubulins from these organisms gave peptide patterns of near identity. Finally, the alpha chains of native tubulins from N. coriiceps neglecta and the cow differed in the sensitivity of their C-terminal domains to digestion by subtilisin. These results demonstrate that the alpha tubulins of Antarctic fishes (but not their beta chains) differ structurally from those of temperate fishes and a mammal.

Animals↗

Clostridium botulinum in fish.

1407 fish caught in Scandinavian waters, the North Sea and the North Atlantic have been examined for the presence of Cl. botulinum. The incidence in gut samples expressed as percentage of fish tested was generally highest in fish from Scandinavian coastal waters and the Baltic Sea (4--43%), decreasing in fish from the North Sea (0--8%), and the organism was practically absent in fish from the North Atlantic. When gut samples were examined, the incidence was highest in demersal fish (cod and flatfish) as compared with pelagic fish (herring). The latter fish species were mainly contaminated on outer surfaces and gills. Only type E was detected in this survey, and Cl. botulinum was not detected in any wild fresh water fish. It is suggested that type E spores may originate in the sea bed and that they be spread by fish and water currents.

Animals↗

[Presence of the bacterial kidney disease in salmonid fish in Quebec (author's transl)].

Presence of the bacterial kidney disease in salmonid fish in Quebec During the summers of 1979 and 1980, wild and hatchery fish were analysed for the presence of the bacterial kidney disease (BKD) agent in salmonid fish in Quebec by the indirect immunofluorescence technique. The causative agent of BKD was detected in all hatcheries tested. Ten to 25% of the fish were positive. The presence of this agent was independent of age and species. We were unable to detect the BKD in fish from the rivers in the northern part of Quebec (over the 50th parallel). The first detection of BKD in Quebec was made in the spring of 1979 in a hatchery. A high mortality occurred in fish over a year old. The majority of the fish were positive by immunofluorescence and the bacteriun was isolated from some fish. Three weeks before the disease appeared, six ponds received fish from this hatchery. No mortality was observed in those ponds and in the middle of the summer the percentage of carrier fish by immunofluorescence was approximately 20%.

Animals↗

Effects of dietary fish oil on serum lipids and VLDL kinetics in hyperlipidemic apolipoprotein E*3-Leiden transgenic mice.

Studying the effects of dietary fish oil on VLDL metabolism in humans is subject to both large intra- and interindividual variability. In the present study we therefore used hyperlipidemic apolipoprotein (APO) E*3-Leiden mice, which have impaired chylomicron and very low density lipoprotein (VLDL) remnant metabolism, to study the effects of dietary fish oil on serum lipids and VLDL kinetics under highly standardized conditions. For this, female APOE*3-Leiden mice were fed a fat- and cholesterol-containing diet supplemented with either 0, 3 or 6% w/w (i.e. 0, 6, or 12% of total energy) of fish oil. Fish oil-fed mice showed a significant dose-dependent decrease in serum cholesterol (up to -43%) and triglyceride levels (up to -60%), mainly due to a reduction of VLDL (-80%). LDL and HDL cholesterol levels were not affected by fish oil feeding. VLDL-apoB kinetic studies showed that fish oil feeding resulted in a significant 2-fold increase in VLDL-apoB fractional catabolic rate (FCR). Hepatic VLDL-apoB production was, however, not affected by fish oil feeding. VLDL-triglyceride turnover studies revealed that fish oil significantly decreased hepatic VLDL-triglyceride production rate (-60%). A significant increase in VLDL-triglyceride FCR was observed (+70%), which was not related to increased lipolytic activity. We conclude that APOE*3-Leiden mice are highly responsive to dietary fish oil. The observed strong reduction in serum very low density lipoprotein (VLDL) is primarily due to an effect of fish oil to decrease hepatic VLDL triglyceride production rate and to increase VLDL-apoB fractional catabolic rate.

Animals↗

Effects of light and presence of fish on lure display and larval release behaviours in two species of freshwater mussels.

We investigated how two sympatric species of freshwater mussels transmit their parasitic larvae to fish hosts. We found that Villosa nebulosa and V. vibex both display large mantle lures to attract potential host fish, but V. nebulosa displayed only at night and V. vibex displayed mostly by day. Display periods were similar in the laboratory and in the field. In two laboratory experiments, we found that the frequency of lure display in both mussel species was unrelated to the presence of fish or to the species of fish present. However, both species released more larvae in the presence of a suitable host fish (Micropterus spp.) and a nonhost species (Cyprinella camura) than in the absence of fish. In all treatments, females released low numbers of larvae on a daily basis throughout the experiment. We also observed several, irregularly occurring major release events in which numbers of larvae released were from one to three orders of magnitude larger than minor, daily releases. In V. nebulosa, major releases occurred with suitable and nonsuitable host species; in V. vibex major releases occurred mostly with suitable host species. In an additional laboratory experiment, we found that V. vibex released large numbers of larvae only when the host fish was able to make physical contact with the mussel. Few larvae were released when no fish were present or when host fish were present but physical access to the mussel was restricted. These results show that, in mussel species that display lures, physical interaction with a fish is necessary to stimulate large releases of larvae and suggest that interactions with a suitable host species stimulate larger and more frequent releases than with nonhosts. Copyright 2000 The Association for the Study of Animal Behaviour.

Journal Article↗

Trophic Transfer of a Sediment-Associated Organophosphate Pesticide from Meiobenthos to Bottom Feeding Fish

Experiments were conducted to examine the dynamics of a sediment-associated pesticide azinphosmethyl (APM) using a sublethal benthic based trophic transfer model (meiobenthic copepods to juvenile fish). Two predominant pathways for contaminant transfer during feeding, prey ingestion and sediment ingestion, were examined to determine their relative contributions to APM transfer and subsequent effects on fish brain AChE inhibition. Experiments were conducted in 1993 and in 1994. Field collected benthic copepods were exposed to 14-C labeled APM, a potent acetylcholinesterase (AChE) inhibitor, in sediments for 96 h. APM burdens were measured in the copepods, and these contaminated copepods were fed to the juvenile fish predator Leiostomus xanthurus in uncomtaminated sediments. After gut clearance, fish were examined for brain AChE activity and APM residues in the liver, heart, gut, muscle, gill, and remains. Similar experiments were conducted in which meals of uncontaminated copepods were fed to spot in APM contaminated sediments, to determine the relative contribution of contaminated sediments to APM transfer. Copepods exposed to APM at a mean sediment concentration of 1223 ng/g dry weight accumulated APM at the level of 2.5 &mgr;g/g dry tissue. Brain AChE activity was significantly depressed (23%) in the 1993 fish fed one meal of contaminated copepods, however there was no significant decline in AChE activity in the fish tested during 1994. APM accumulation in fish feeding in contaminated sediments was generally greater than in fish feeding on contaminated prey. Significant accumulation was found in bodily remains, gills, gut, and muscle in fish that fed in contaminated sediments. No significant APM accumulation was found in fish fed the contaminated copepods.

Journal Article↗

Effects of cooking on radiocesium in fish from the Savannah River: exposure differences for the public.

Understanding the factors that contribute to the risk from fish consumption is an important public health concern because of potential adverse effects of radionuclides, organochlorines, other pesticides, and mercury. Risk from consumption is normally computed on the basis of contaminant levels in fish, meal frequency, and meal size, yet cooking practices may also affect risk. This study examines the effect of deep-frying on radiocesium (137Cs) levels and risk to people fishing along the Savannah River. South Carolina and Georgia have issued consumption advisories for the Savannah River, based partly on 137Cs. 137Cs levels were significantly higher in the cooked fish compared to the raw fish on a wet weight basis. Mean 137Cs levels were 0.61 pCi/g (wet weight basis) in raw fish, 0.81 pCi/g in cooked-breaded, and 0.99 pCi/g in cooked-unbreaded fish. Deep-frying with and without breading resulted in a weight loss of 25 and 39%, while 137Cs levels increased by 32 and 62%, respectively. Therefore, the differences were due mainly to weight loss during cooking. However, the data suggest that risk assessments should be based on cooked portion size for contaminant analysis, or the risk from 137Cs in fish will be underestimated. People are likely to estimate the amounts of fish they eat based on a meal size of the cooked portion, while risk assessors determine 137Cs levels in raw fish. A conversion factor of at least two for 137Cs increase during cooking is reasonable and conservative, given the variability in 137Cs levels. The data also suggest that surveys determining consumption should specifically ask about portion size before or after cooking and state which was used in their methods.

Animals↗

Methylmercury, total mercury and total selenium in four common freshwater fish species from Ya-Er Lake, China.

Snakehead fish (Ophiocephalus argus cantor), silver carp (Hypophthalmichthys molitrtix), crucian carp (Carassius carassius), and common carp (Cyprinus carpio) are four common freshwater fish species in China. In this study, the level of methylmercury (MeHg), total mercury (T-Hg), and total selenium (T-Se) in muscle samples of these four fish species from Ya-Er Lake, China, were analyzed using atomic fluorescence spectrometry coupled with high-performance liquid chromatography, and inductively coupled plasma mass spectrometry. The concentrations of MeHg in all the fish species were significantly correlated with those of T-Hg. Higher T-Hg and MeHg concentrations had accumulated in the snakehead fish, which is a strongly predatory fish, than in the other three species. The concentration ratios of MeHg and T-Hg in the muscles of these four fish species were almost equal. Conversely, there was negative correlation between the concentrations of T-Hg and T-Se, which implies that there is a competition between these two elements with respect to bioaccumulation. It is noteworthy that of all the muscle samples tested, the level of T-Hg exceeded the maximum allowable limit in fish [0.4 mg kg(-1) (w/w) recommended by the World Health Organization] in 38.46% of those of the silver carp, 18.18% of those of the crucian carp, and 100% of those of snakehead fish. These results show that the consumption of contaminated fish is a potential threat to human health and that necessary preventive measures to safeguard public health should be emphasized.

Animals↗

Controlled trial of fish oil for regression of human coronary atherosclerosis. HARP Research Group.

OBJECTIVES: This randomized clinical trial tested whether fish oil supplements can improve human coronary atherosclerosis. BACKGROUND: Epidemiologic studies of populations whose intake of oily fish is high, as well as laboratory studies of the effects of the polyunsaturated fatty acids in fish oil, support the hypothesis that fish oil is antiatherogenic. METHODS: Patients with angiographically documented coronary heart disease and normal plasma lipid levels were randomized to receive either fish oil capsules (n = 31), containing 6 g of n-3 fatty acids, or olive oil capsules (n = 28) for an average duration of 28 months. Coronary atherosclerosis on angiography was quantified by computer-assisted image analysis. RESULTS: Mean (+/- SD) baseline characteristics were age 62 +/- 7 years, plasma total cholesterol concentration 187 +/- 31 mg/dl (4.83 +/- 0.80 mmol/liter) and triglyceride levels 132 +/- 70 mg/dl (1.51 +/- 0.80 mmol/liter). Fish oil lowered triglyceride levels by 30% (p = 0.007) but had no significant effects on other plasma lipoprotein levels. At the end of the trial, eicosapentaenoic acid in adipose tissue samples was 0.91% in the fish oil group compared with 0.20% in the control group (p < 0.0001). At baseline, the minimal lumen diameter of coronary artery lesions (n = 305) was 1.64 +/- 0.76 mm, and percent narrowing was 48 +/- 14%. Mean minimal diameter of atherosclerotic coronary arteries decreased by 0.104 and 0.138 mm in the fish oil and control groups, respectively (p = 0.6 between groups), and percent stenosis increased by 2.4% and 2.6%, respectively (p = 0.8). Confidence intervals exclude improvement by fish oil treatment of > 0.17 mm, or > 2.6%. CONCLUSIONS: Fish oil treatment for 2 years does not promote major favorable changes in the diameter of atherosclerotic coronary arteries.

Adipose Tissue↗

Changes in microcystin production by Microcystis aeruginosa exposed to phytoplanktivorous and omnivorous fish.

With direct exposure to phytoplanktivorous fish (Hypophthalmichthys molitrix), increased mass-specific microcystin production occurred in three monoclonal Microcystis aeruginosa strains (NIES 44, 88 and 99). Total mass-specific microcystin content of NIES 44 exposed to H. molitrix was over 50 times higher than controls (a mean value of 16.2 microgg(-1)-dry cell in controls versus 878.6 microgg(-1)-dry cell in treatments). Up to nine times higher microcystin levels were detected in NIES 88 exposed to H. molitrix compared to controls (a mean value of 553 in controls versus 5145 microgg(-1)-dry cell in treatments). The microcystin levels of all strains were significantly different between controls and H. molitrix treatments (P < 0.01 for NIES 44 and 88; P < 0.05 for NIES 99). The microcystin response to the omnivorous Carassius gibelio langsdorfi was weaker than that of H. molitrix, though the levels in all strains exposed to the fish were higher than in controls and a significant difference in microcystin production between controls and omnivorous fish treatments occurred for NIES 44 (a mean value of 6.9 in controls versus 41.5 microgg(-1)-dry cell in treatments; P < 0.01) and NIES 88 (a mean value of 359.8 versus 480.4 microgg(-1)-dry cell; P < 0.05). Microcystis cells were observed in the both fish faeces and gut contents, and microcystin was also detected in the body tissues (from 0.6 to 2.5 microgg(-1)-dry weight) and faeces of both fish species on the final day of experiment, although 98% of fish in three strains of Microcystis cultures had lost weight (mean +/- S.E. fish growth rate with M. aeruginosa; -0.90 +/- 0.06% per day, n = 96). This study showed that several M. aeruginosa strains increased toxin production when exposed to fish, especially phytoplanktivorous species, even though fish appeared not to feed vigorously on toxic Microcystis, and supports the hypothesis that this response is a fish-induced defence mediated by physical contact associated with feeding or by chemical cues (e.g. kairomones).

Analysis of Variance↗

Development of chronic tests for endocrine active chemicals. Part 1. An extended fish early-life stage test for oestrogenic active chemicals in the fathead minnow (Pimephales promelas).

An extended early-life stage test (based on OECD test guideline 210) was developed to allow the evaluation of a weak environmental oestrogen, 4-tert-pentyphenol (4TPP), on sexual differentiation and gonadal development. Fathead minnow (Pimephales promelas) embryos were exposed to three concentrations of 4TPP (56, 180 and 560 microg l(-1)) in a flow-through system, at 25+/-1 degrees C, for <107 days post-hatch (dph). In addition, some embryos were exposed to 180 microg 4TPPl(-1) until 30 or 60 dph, after which they were exposed to dilution water only until 107 dph. At 30, 60 and 107 dph fish were evaluated for growth and gonadal development (via histology), and at 107 dph fish were also evaluated for secondary sexual characteristics (SSC), gonadosomatic index (GSI) and plasma vitellogenin (VTG). There were no effects of 4TPP on hatching success or survival, however, there was a delay in the time taken for embryos to hatch (560 microg 4TPPl(-1)). No treatment-related effects were observed on fish growth, with the exception of at 107 dph when the condition factor in female fish was reduced in all 4TPP continuous exposure treatments. Plasma VTG was only elevated in female fish exposed to 180 microg 4TPPl(-1) and inhibition of gonadal growth (GSI) occurred only in females exposed to 560 microg 4TPPl(-1). Histological examination of the gonads revealed delays and disruption in male sexual differentiation and development (180 microg 4TPPl(-1)) and no testicular tissue was observed in any fish exposed to 560 microg 4TPPl(-1). Mixed gonads (predominately testes with a scattering of primary oocytes) were present in fish exposed to all doses of 180 microg 4TPPl(-1) at 107 dph. Feminisation of the reproductive ducts (formation of an ovarian like cavity) occurred in the testis of all males exposed to 180 microg l(-1), regardless of length of 4TPP exposure. Results indicate that the period of 30-60 dph appears to be the sensitive window for disruption of formation of the reproductive duct and this effect is not reversible when the fish are transferred to dilution water. The data also show that this integrative test is suitable for the detection of a weak environmental oestrogen and comparisons of these results with that of a fish full life-cycle, in medaka, indicate that this test could be a suitable surrogate for a fish full life-cycle.

Animals↗

Development of chronic tests for endocrine active chemicals. Part 2: an extended fish early-life stage test with an androgenic chemical in the fathead minnow (Pimephales promelas).

The Endocrine Modulators Study Group (EMSG) of the European Chemical Industry has proposed an extended fish early-life stage (ELS) test based on OECD test guideline 210 in combination with a fish pair-breeding reproduction study as a possible alternative for fish full life cycle testing. In this paper the androgen methyldihydrotestosterone (MDHT) was tested in an extended ELS test with fathead minnow supplementary to such a test with the weak estrogen 4-tert-pentylphenol (4TPP). Main endpoints were secondary sexual characteristics (SSC), plasma vitellogenin (VTG) induction and gonadal development. Early blastula embryos were exposed to 0, 0.10, 0.32 and 1.0 microgMDHTl(-1) for up to 114 days post-hatch (dph). A batch of fish exposed to 1.0 microg l(-1) was transferred to clean water after 30 or 63 dph for the remainder of the study. Ethinylestradiol (EE2) was included as estrogenic reference substance at 0.01 microg l(-1). Exposure to MDHT had no significant effect on hatching success or survival, but significantly increased the condition factor of fish exposed for 63 and 114 dph (up to 150% of the control). At 63 dph MDHT exposure induced appearance of tubercles on the snout (a male SSC) of more than 80% of fish. Compared to the controls, plasma VTG was not detectable or significantly lower in fish exposed to MDHT at 0.10 microg/l, but not significantly affected at higher MDHT concentrations. Both lower levels of MDHT significantly inhibited the development of female gonads as of 30 dph. Fish exposed to MDHT at 0.32 and 1.0 microg l(-1) showed higher incidences of mixed sex gonads (10-25%) and smaller testes or dysplasia of gonadal tissue. Dysplasia was present in 80% of the fish continuously exposed to 1.0 microg l(-1) up to 114 dph, but reversible when fish were transferred to dilution water. Results indicate that suppression of ovarian development was the most sensitive endpoint for MDHT exposure after 30 dph. Other endpoints (e.g., growth and SSC) required exposure during at least up to 63 dph to yield a significant effect. Androgenic effects on VTG production required even longer exposure, i.e., until sufficient number of females had matured.

Androgens↗