Depression of feeding and growth rates of the seastar Evasterias troschelii during long-term exposure to the water-soluble fraction of crude oil

To test the effect of petroleum hydrocarbons on predation by the seastar Evasterias troschelii (Stimpson, 1862) on the mussel Mytilus edulis (L.), we exposed the predator with the prey to six concentrations of the water-soluble fraction (WSF) of Cook Inlet crude oil. Seastars and mussels were collected at Auke Bay, Alaska, in November 1980. During a 28 d exposure in a flow-through system, seastars were more sensitive to the WSF than mussels: the LC₅₀ for the seastars was 0.82 ppm at Day 19 and, although no mussels were exposed to WSF for more than 12 d, none died. Daily feeding rates (whether in terms of number of mussels seastar^-1 d^-1 or dry weight of mussels seastar^-1 d^-1) were significantly reduced at all concentrations above 0.12 ppm. At 0.20, 0.28 and 0.72 ppm WSF, daily feeding rates (in terms of dry weight of mussels) were, respectively, 53, 37, and 5% of the control rate; at the two highest concentrations (0.97 and 1.31 ppm WSF), the seastars did not feed. Seastars at concentrations greater than 0.12 ppm WSF grew slower than individuals from the control group and the 0.12 ppm-treatment group combined. These laboratory results show that E. troschelii is more sensitive to chronic low levels of the WSF of crude oil. The possibility that such oil pollution could reduce predation and permit M. edulis to monopolize the low intertidal zone of southern Alaska remains to be studied.     

Individual and combined toxicity of some petroleum aromatics to the marine amphipod Elasmopus pectenicrus

Toxicity of 4 components of petroleum oils to the marine amphipod Elasmopus pectenicrus (Bate) has been assessed. Two ephemeral aromatic hydrocarbons, naphthalene (A) and 1, 2, 4-trimethylbenzene (B) were more toxic than two persistent aromatics, o-cresol (C) and o-toluidine (D). The acute toxicity concentrations obtained for individual aromatic compounds were always greater than the actual concentrations found in the water-soluble fractions (WSF) of fuel oils. Results from mixtures of 2 or more components indicated that the LC₅₀ levels were primarily determined by the more toxic substances, A and B. Naphthalene and 1, 2, 4-trimethylbenzene became more toxic to the E. pectenicrus when present in a mixture of more than 2 components, and the toxicity increased with increasing numbers of components present. Synergistic effects, therefore, possibly occur in the whole WSF. No antagonistic effects were observed among the 4 petroleum aromatics.University of Texas, Marine Science Institute Contribution No. 290     

Ventilatory responses of the shrimp Palaemon adspersus to sublethal concentrations of crude oil extract

The shrimp Palaemon adspersus Rathke was exposed to the water soluble fraction (WSF: 50; 100; 200 ppb) of North Sea crude oil. The ventilatory behaviour, measured with impedance techniques, was followed for 2 wk and in a subsequent recovery period for 5 wk. Exposure to WSF caused a gradual increase in arrhythmic scaphognathite activity. After 6 d of exposure to 200 ppb shrimps spent more than 50% of the time beating arrhythmically, persisting and sometimes increasing to 100% during the remainder of the exposure period. In the 100 and 50 ppb groups, the proportion of time spent beating arrhythmically increased gradually with exposure time reaching 30 and 20% respectively. After 4 wk in clean seawater no signs of recovery were evident for individuals previously exposed to 200 ppb while those previously in 50 ppb had all recovered. After 5 wk those previously in 100 ppb had recovered and the 200 ppb group showed signs of recovery. The disturbances in ventilatory behaviour when exposed to WSF is considered to be due to damage to gill membranes or to nerve tissue, especially to the neurons in the suboesophageal ganglion controlling scaphognathite activity.Supported by the Environmental Research Programme under the Commission of European Communities (Contribution No. ENV/350/DK (G))     

Role of adipocytes in the muscle tissue of Atlantic salmon (Salmo salar ) in the uptake, release and retention of water-soluble fraction of crude oil hydrocarbons

The uptake and depuration of the water-sol‐uble fraction (WSF) of hydrocarbons of crude petroleum by Atlantic salmon (Salmosalar) has previously been examined in terms of whole muscle. The hypothesis that the tainting WSF in the muscle was retained primarily by adipocytes has been investigated by the isolation of adipocytes and the subsequent analysis for hydrocarbons in adipocytes. After 96 h exposure of market-sized Atlantic salmon to 0.2 ppm WSF, adipocytes isolated from the belly flap region of the muscle tissue accumulated 14.3 times more WSF (59.4 ppm) than the dorsal white muscle (4.2 ppm), while 54% of the tainting WSF in the dorsal white muscle was found to be stored in associated adipocytes. When returned to clean seawater, WSF accumulated in the dorsal white muscle was released much faster than that in the adipocytes. These results indicated that the loose association of WSF with the nonlipid portion of white muscle, mainly muscle cells and intercellular fluid, is responsible for the rapid discharge of WSF from the dorsal muscle tissue in the early stages of depuration. After 4 d of depuration, the adipocytes became the principal storage site of residual WSF in white muscle and the depuration of WSF from muscle tissue then reflected the release of WSF from adipocytes in the muscle tissue. After 20 d of depuration, 10.7 ppm of tainting WSF in the form of high molecular weight aromatic hydrocarbons (mainly C₄-benzenes, naphthalene and alkylated naphthalenes) were still present in adipocytes, while in the dorsal white muscle only a trace of total WSF was detected. Increases in the number of aromatic rings and the alkylations on the rings enhanced the accumulation and retention of individual hydrocarbons in both adipocytes and white muscle. From these studies we conclude that it is the adipocytes in the muscle tissue which control the actual accumulation and release of hydrocarbons in the whole muscle tissue of Atlantic salmon. Received: 21 August 1996 / Accepted: 26 September 1996     

Occurrence of tri-n-butyltin-caused imposex in the North Pacific marine snail Nucella lima in Auke Bay,Alaska

Like female dog whelks (Nucella lapillus) in the Atlantic Ocean, females of the Pacific gastropod N. lima respond to low concentrations of tri-n-butyltin (TBT) by growing a penis and vas deferens. This condition, termed imposex, was found to occur in N. lima collected from August 1987 to May 1988 along a TBT pollution gradient associated with a marina in Auke Bay, Alaska. The suite of symptoms characteristic of imposex in N. lima was slightly different than in N. lapillus. Imposex, as measured by relative penis size (RPS) of females to males, increased from 0.0 to 34.27 along this gradient and as measured by vas deferens sequenceindex (VDS) increased similarly from 0.0 to 4.29. Concentrations of TBT in N. lima increased from below detection limits (about 0.010 g Sn g^-1 dry tissue wt) to 0.065 g Sn g^-1 dry tissue wt along the gradient. The gradient was determined by measuring TBT in whole-body tissues of bay mussels (Mytilus edulis). Concentrations of TBT in mussels increased from below detection limits to 0.833 g Sn g^-1 dry tissue wt in mussels from within the marina that was the major local source of TBT. Imposex, tissue TBT burden, and position along a TBT pollution gradient are significantly correlated (P<0.01) in N. lima. TBT was tested as a causative agent for imposex by exposing snails at a distant control site to TBT paint. After 1 mo exposure, 33% of the females grew a penis ranging in size from 0.2 to 0.8 mm. Our results generally corroborate those found for N. lapillus and indicate that imposex in N. lima is caused by environmental TBT exposure. We suggest that the RPS in the genus Nucella may be useful in monitoring TBT in coastal waters worldwide.     

Characteristics of dispersions and water-soluble extracts of crude and refined oils and their toxicity to estuarine crustaceans and fish

The quantitative hydrocarbon composition and behavior in seawater of water-soluble fractions (WSF) and oil-in-water dispersions (OWD) of 4 oils was investigated. Two crude oils, South Louisiana crude and Kuwait crude, and two refined oils, No. 2 fuel oil and bunker C residual oil, were used in these investigations. The WSFs of the crude oils had higher total oil-hydrocarbon concentrations and were richer in light aliphatics and single-ring aromatics than were the WSFs of the refined oils. The WSFs of the refined oils contained significantly higher concentrations of naphthalenes than did those of the crudes. The hydrocarbon composition of the aqueous phase of OWDs closely resembled that of the parent oils. Gentle aeration of the OWDs resulted in a loss of 80 to 90% of the aqueous hydrocarbons in 24 h. Alkanes disappeared from the dispersions more rapidly than aromatics. The WSFs and OWDs of the refined oils were considerably more toxic to the 6 test species than were those of the crude oils. The test species can be ranked according to increasing sensitivity to oil as follows: Cyprinodon variegatus, Menidia beryllina, Fundulus similus, Penaeus aztecus postlarvae, Palaemonetes pugio and Mysidopsis almyra. The results of this investigation are discussed in relation to the potential impacts of oil spills on the marine and estuarine environments.     

Uptake and Dupuration of Petroleum Hydrocarbons By the Backwater Clam,Meretrix Casta Chemnitz

Meretrix casta were experimentally exposed to water soluble fractions of refined and crude oil and their rate of accumulation of the petroleum hydrocarbons including total and individual aromatics viz., naphthalene, methylnaphthalene and dimethylnaphthalene, was studied. Subsequent transfer to clean waters in the laboratory and field resulted in depuration of the accumulated hydrocarbons from tissues. in general, the rate of discharge was found to be dependent on the concentration that the animals had been earlier exposed to and also the alkylation of aromatic hydrocarbons. Occasional increases were observed in the hydrocarbon values of clams which were placed in the field, compared to their laboratory counterparts, suggesting an intermittent source of petroleum hydrocarbon input into the environment. the influence on the rate of uptake, release and retention of hydrocarbons in the clams is discussed.     

Hydrocarbons in benthic algae from the eastern Gulf of Mexico

Thirty-six samples of benthic algae were collected from the continental shelf along the eastern Gulf of Mexico. The algae contain 82.8±143 ppm aliphatic hydrocarbons by dry weight and 11.8±22.7 ppm aromatic and polyolefinic hydrocarbons. The aliphatic constituents of red algae are composed almost entirely of n-C₁₇ (70 to 95% of aliphatic weight); green algae have varying amounts of n-alkanes in the range of n-C₁₅ to n-C₁₉, with homologous series of odd carbon number n-alkenes. Phytadienes occur as high as 740 ppm dry weight in the green algae and do not appear to be related to time or place of collection or to presence of any petroleum pollutants. The single brown algal species contains predominantly n-C₁₅. About 30% of the samples have hydrocarbons resembling degraded petroleum residues, but no petroleum pollution of recent origin was detected in any specimen.     

Effects of water-soluble fraction of the Mexican crude oil “Isthmus Cactus” on growth,cellular content of chlorophylla,and lipid composition of planktonic microalgae

Phytoplankton species were grown in batch cultures in the presence of the water-soluble fraction (WSF; 50 and 100%) of a Mexican crude oil (Isthmus Cactus). The algae exhibited various responses ranging from retarded growth to stimulation of growth. The cellular content of chlorophylla and the lipid composition of the algae were examined. Four algae, the bacillariophytesNitzschia closterium andAsterionella glacialis, the cryptophytesRhodomonas lens, and the chlorophyteDunaliella tertiolecta, exhibited retarded growth. In most of these algae, cellular chlorophylla, lipid pigments, glycolipids and triglycerides decreased whereas sterols and hydrocarbons accumulated. Phospholipids did not exhibit any specific pattern of change during the experiments. The cyanophyteAgmenellum quadruplicatum and the bacillariophyteSkeletonema costatum were less sensitive to the WSF. The cell yield of the dinophyteProrocentrum minimum was stimulated by the WSF. In these three latter species, lipid pigments were enhanced or remained at control levels. We concluded that the toxic effect of the WSF disrupts the biosynthesis mechanisms required for a functional photosynthetic apparatus (biosynthesis of chlorophylla, glycolipids and lipid pigments) in sensitive algae, a phenomenon coupled to sterol accumulation in these algae.     

Biochemical responses of the striped mullet Mugil cephalus to oil exposure I. Acute responses—Interrenal activations and secondary stress responses

Acute single exposures to the water-soluble fraction (WSF) of a No. 2 fuel oil influences several biochemical parameters in juveniles of Mugil cephalus Linnaeus. Plasma cortisol and glucose concentrations were measured at 1 and 3 h after exposure to 1, 5, 10 or 20% WSF. No elevation of plasma cortisol or glucose levels occurred in fish exposed to the lowest concentration of oil, whereas a dose-response relationship was observed at higher doses. The dynamics of plasma corticosteroid, glucose and cholesterol concentrations and osmolality as well as accumulation of naphthalenes in the fish tissues were monitored during exposure to 20% WSF. Circulating cortisol concentrations rose rapidly to 5 times normal values 1h after exposure to oil and subsequently declined to control levels 6 h after oil addition. A smaller secondary rise occurred at 12 h, but cortisol had returned to basal levels 12 h later. In contrast, plasma glucose, cholesterol and osmolality rose more slowly to reach maximum values between 3 and 4 h after oil addition. By 24 h plasma cholesterol and osmolality had returned to normal values whereas the hyperglycemia persisted. However, 72 h after the addition of WSF all biochemical parameters had returned to control levels. At this time considerable accumulation of total naphthalenes had occurred in several fish tissues, whilst the concentration of total naphthalenes in the exposure tanks had declined to background levels. When freshly prepared 20% WSF was added to the exposure tanks during this period, all biochemical parameters were again elevated. The results suggest that the volatile components of fuel oil in the water trigger the biochemical changes described in a dosedependent manner. The possible ecological significance of these changes and the potential use of these parameters as sublethal indicators of environmental contamination are discussed.     

Formation of metallothioneins in relation to accumulation of cadmium in the common mussel Mytilus edulis

Accumulation rates of Cd have been studied in the common mussel Mytilus edulis L. under different laboratory conditions. Semi-static and through-flow experiments were carried out at Cd concentrations of 200 ppb and 50/100 ppb in the water, respectively. A linear uptake of Cd throughout the experimental periods of 36 or 124 d was found in all experiments. The accumulation rate was 3.10 ppm d^-1 at 50 ppb in the through-flow water experiment, 4.1 ppm d^-1 in starved and 6.6 ppm d^-1 in fed mussels in the semi-static experiments with 200 ppb Cd in the water. The Cd-accumulation rate in various fractions of soft parts decreased in the order: body>mantle>muscles. Elution patterns obtained from gel-filtrations showed an increasing amount of Cd-binding proteins (metallothioneins) when the body burden of Cd increased. The Cd content in the body fraction of mussels transferred to clean water for 42 d after first being exposed to Cd during 124 d decreased from 564 ppm on a dry weight basis to 417 ppm while the fraction of Cd bound to metallothioneins rose from 22 to 78% during the same period.     

Short-term effect of microencapsulated hydrocarbons on shell growth of Mytilus edulis

Two North Sea crude oils, weathered crude oil, n-alkanes, medical liquid paraffin and a fish oil were microencapsulated and added to seawater. This gave a medium containing both water soluble fractions (WSF) and oil particles, and thus simulated the conditions in natural seawater contaminated with oil. The microencapsulated oils were fed to Mytilus edulis L. in different concentrations, and the growth in terms of shell length of the mussels was measured in intervals of 24 to 48 h for 4 to 12d. With the crude oilss A and B, weathered crude oil and n-alkanes at concentrations1 mg l^-1, shell growth rate decreased rapidly compared to controls. With liquid paraffin at levels of 1 to 12 mg l^-1, a small but significant negative effect on shell growth occurred after 5 d of exposure. Exposure to fish oil at 1 and 4 mg l^-1 gave no significant reduction in growth rate. The toxicity of the different hydrocarbons was not related to their content of aromatic fractions. Crude oil B was tested at concentrations ranging from 0.12 to 12 mg l^-1. At 0.12 mg l^-1 shell growth was not significantly different from the control, while at 0.25 and 0.50 mg l^-1 a temporary and significant stimulation of growth was observed. The product concentration (C)xresponse (R) shows a linear regression on exposure time (t). The regression model CxR=75-0.18t is used to estimate EC-values (effect of a given concentration) for given response levels. The model gives a very good fit to observed data.     

Chemical composition and effects of water extracts of petroleum on eggs of the sand dollar Melitta quinquiesperforata

Sperm and eggs of sand dollars, Melitta quinquiesperforata (Leske), were subjected to two petroleum oils, and effects determined. The oils chosen were Kuwait crude and No. 2 fuel oil, supplied by the American Petroleum Institute. Water-soluble extracts (WSF) from oil-sea water mixes were prepared and the major aromatic components in the WSF of the fuel oil were identified. WSF of No. 2 fuel oil depressed respiration, mobility of sperm, interfered with fertilization and cleavage, and retarded larval development. The effects were detectable at dilutions of 4% and less (about 0.6 ppm of WSF). Kuwait crude was much less toxic. There was no effect on water permeability of the egg membrane. Results are compared with similar studies on other marine organisms.     

Accumulation of cadmium and bioenergetics in the mussel Mytilus edulis

Accumulation rates of cadmium, the amount of food ingested and assimilated, the amount of oxygen consumed and changes in dry flesh weight have been measured in Mytilus edulis L. exposed to 0, 10 and 100 ppb cadmium for 17 d in aquaria with seawater flowing continously and at constant algal concentration. The accumulation rates were linear at 10 and 100 ppb, amounting to 0.58 and 8.89 ppm d^-1, respectively. Body loads up to 150 ppm caused no effects on either clearance, ingestion, assimilation, respiration, or growth. High net growth efficiencies between 55–59% were obtained, indicating near optimal experimental conditions. It is suggested that the setup and experimental procedure provide an excellent tool in the study of accumulation and sublethal effects of environmental pollutants in suspension feeding bivalves.     

Very long-chain aliphatic hydrocarbons in lipids of mussels (Mytilus edulis) suspended in the water column near petroleum operations off Sable Island,Nova Scotia,Canada

Mussels (Mytilus edulis) suspended in the water column in 1994 and 1995 for the monitoring of oil drilling operations off Sable Island, Nova Scotia were examined for hydrocarbon profiles, particularly aliphatic hydrocarbons. A spring bloom of phytoplankton occurred during the 90-d suspension period in 1995. Hydrocarbons isolated from the 1995 suspended mussels showed very high concentrations of both biogenic hydrocarbons and very long-chain n-alkanes from C₂₀ to C₃₂, initially thought to be petrogenic. Both types of hydrocarbons were either not detected or were only present in trace amounts in the mussels suspended in 1994 at similar sites. The biogenic hydrocarbons in the 1995 mussels were apparently of planktonic origin, from the spring bloom, and were dominated by heneicosahexaene (21:6), followed by pristance, heptadecane, and varions monounsaturated and polyunsaturated phytenes, heptadecenes, nonadecenes and heneicosenes. They could be readily hydrogenated to yield the basic alkanes. The 1995 mussels suspended within 1 km from the oil well platform were probably slightly tainted by petrogenic hydrocarbons, as evidenced by the detection of phytane and high concentrations of total aliphatic hydrocarbons, whereas the mussels suspended 10 km from the platform showed only high concentrations of biogenic hydrocarbons and the novel long-chain n-alkanes. The occurrence of an unusual phytoplankton bloom during the suspension period severely interfered with the petroleum monitoring role of mussels by altering the mussel hydrocrbon profiles through the accumulation into and probably selective depuration of xenobiotic hydrocarbons from the mussel, tissues.J. Parsons (deceased)     

A long-term study of the effects of water-soluble fractions of No. 2 fuel oil on the survival,development rate,and growth of the mud crab Rhithropanopeus harrisii

Mud crabs, Rhithropanopeus harrisii (Gould), were exposed continuously for 6 months after hatching to water-soluble fractions (WSF) of No. 2 fuel oil. Survival, growth and development rate were monitored during this time. The zoeal stages were the most sensitive to fuel oil. A 20% WSF (0.36 ppm total naphthalenes, 1.26 ppm total hydrocarbons) was acutely toxic to these stages. Of the zoeal stages, the first stage appeared to be the most sensitive. The combined duration of the 4 zoeal stages was significantly increased by increasing WSF exposure concentrations. The megalopa and crab stages were not particularly sensitive to continued petroleum hydrocarbon exposure, particularly when compared to zoeal stages. However, mean duration of the megalopa and first crab stages was significantly affected by oil exposure. Individuals which survived the highest exposure concentrations as larvae appeared to grow larger during the crab stages, so that at the end of 6 months comparably staged crabs were equal to or larger than both control crabs and those exposed to low WSF concentrations. Stage distributions at the end of 6 months showed no differences due to WSF exposure. Sex ratios, which could be determined at the end of 6 months, were approximately 1, indicating no sex-related differential sensitivity to WSF exposure, at least as larvae or juveniles. The data indicate that these crabs possess considerable ability to recover from the effects of chronic sublethal exposure to petroleum hydrocarbons. The most deleterious effects of oil pollution on this species may be due to its impact on larval recruitment into the adult population.     

Responses of Mytilus edulis on exposure to the water-accommodated fraction of North Sea oil

Individuals of Mytilus edulis L., collected from the Erme estuary (S.W. England) in 1978, were exposed to low concentrations (7 to 68 g l^-1) of the water-accommodated fraction (WAF) of North Sea crude oil. The pattern of accumulation of petroleum hydrocarbons in the body tissues was affected by the presence of algal food cells, the period of exposure, the hydrocarbon concentration in seawater, the type of body tissue and the nature of the hydrocarbon. Many physiological responses (e.g. rates of oxygen consumption, feeding, excretion, and scope for growth), cellular responses (e.g. lysosomal latency and digestive cell size) and biochemical responses (e.g. specific activities of several enzymes) were significantly altered by short-term (4 wk) and/or long-term (5 mo) exposure to WAF. Stress indices such as scope for growth and lysosomal latency were negatively correlated with tissue aromatic hydrocarbons.     

Changes in the digestive gland of Euphausia superba during short-term starvation: lipid class,fatty acid and sterol content and composition

During a period of short-term (19 d) starvation, total lipid in the digestive gland of Euphausia superba Dana decreased from 21 to 9% dry weight. Total lipid per digestive gland decreased significantly during starvation compared to Day 0 individuals, falling from 1960 (±172) to 385 (±81) g. Polar lipid was the major lipid class utilised during starvation, falling from 1510 (±225) to 177 (±46) g per digestive gland (76 to 45%). Absolute levels of triacylglycerol fell from 300 (±41) to 76 (±5) g; however, relative levels remained unchanged. The relative level of free fatty acid increased significantly with starvation (4 to 39%) with absolute levels ranging from 79 (±1) to 156 (±20) g per digestive gland. Absolute levels of all fatty acids per digestive gland declined continually until the end of the starvation period. The long-chain polyunsaturated acids eicosapentaenoic (20:53) and docosahexaenoic (22:63), decreased with starvation from 37 to 26% and 15 to 10%, respectively whereas the saturated fatty acid, palmitic acid (16:0), increased from 15 to 20%. Cholesterol, the major sterol in this organ, increased from 17 (±20) to 44 (±13) g per digestive gland by Day 3, and by Day 19 had returned to levels found in the digestive gland of Day 0 individuals. Desmosterol followed a similar pattern to cholesterol, increasing from 3 (±1) g per digestive gland on Day 0 to 11 (±4) g on Day 3, and falling to 2 (±1) g on Day 19. Other sterols in the digestive gland, predominantly of algal origin, fell from the levels found in Day 0 individuals to near zero amounts by Day 6. The digestive gland of E. superba plays a dynamic role during shortterm starvation in terms of lipid content and composition. The relative levels of polar lipids, free fatty acids and cholesterol in the digestive gland may provide reliable indices of the nutritional condition of E. superba in the field. Sterols in the digestive gland are indicative of recent dietary composition of krill, and may also be used to quantify dietary input from individual phytoplanktonic species.     

Effects of benzo(k)fluoranthene,a polycyclic aromatic hydrocarbon on DNA damage,lipid peroxidation and oxidative stress in marine gastropod Morula granulata

Benzo(k)fluoranthene [B(k)F] is one of the widespread priority pollutants of polycyclic aromatic hydrocarbons that has been scarcely studied for exposure assessment. With studies reporting a high amount B(k)F in sediments and water samples around the world, it has become vital to study its effects on aquatic organisms. In this connection, this study is conducted to study the effect of different concentrations of B(k)F (1, 10, 25 and 50?µg/L) in marine gastropod Morula granulata exposed in vivo for 96?h. A concentration-dependent increase in percentage tail DNA (TDNA) as measured by comet assay was observed in snails exposed to B(k)F. Exposure concentrations above 1?µg/L B(k)F showed significant increase in superoxide dismutase (SOD) activity and lipid peroxidation value in snails. After 96?h, SOD activity was found to be doubled for 50?µg/L B(k)F in comparison to control. A significant increase in catalase and glutathione S-transferase activity was observed at all exposure conditions at the end of the exposure time. Our study showed that B(k)F induces oxidative stress in snails which further lead to genotoxic damage. To our knowledge, this is the first study on oxidative stress and genotoxic damage in gastropods exposed to B(k)F.     

Monitoring toxicity of an azo dye methyl red and a heavy metal Cu,using plant and animal bioassays

Toxicities of an azo dye methyl red and a heavy metal copper (Cu) were quantified, using growth and mortality as end points, in four plant species and three animal species by subjecting them to short-term (4 days for animals, 10 days for plants) static bioassays. Lemna aequinoctialis Welwitch (EC₅₀: 7–16 ppm) was found to be the most sensitive species for methyl red, Ceratophyllum demersum L. (EC₅₀: 25 ppm) and Lactuca sativa L. (EC₅₀: 56 ppm) were intermediate, while Oryza sativa L. shows reduction in seedling vigor (9–27%) of <50%, being the least sensitive amongst the tested plant species. Methyl red toxicity is almost 3–5-fold higher in growing medium (pH = 5.8–6.0), even at high nutrient levels, while Cu toxicity is higher in nutrient-poor alkaline medium at alkaline pH (8.3–8.7; EC₅₀: Ceratophyllum = 104–200 ppb; Lemna = 100–170 ppb) compared to nutrient-rich acidic medium (pH = 5.4–5.7; EC₅₀: Ceratophyllum = 2600–3175 ppb; Lemna = 4350–4715 ppb). Rice tolerance (EC₅₀: 6500 ppb) was found to be higher than hydrophytes while lettuce was most tolerant to Cu. Fish sensitivity toward the test chemicals was almost parallel to Ceratophyllum and Lemna [Gambusia affinis Baird and Gerard (LC₅₀: 250 ppb for Cu) and Poecilia reticulata Peters (LC₅₀: 24 ppm for methyl red)]. Similar to the plants, dye toxicity increased markedly (LC₅₀: 7 ppm) in the acidic medium (pH = 6.0). Amongst the tested organisms, Daphnia was found to be most sensitive to methyl red (EC₅₀: 6 ppm) while its sensitivity to Cu (EC₅₀: 230 ppb) was similar to hydrophytes and fish. Initially, the combination of dye and Cu (at their sublethal concentrations) had additive effects in duckweed, while dye concentration ruled afterward. These results indicate that hydrophytes and animals are equally sensitive toward the test chemicals. Dye toxicity in hydrophytes and fish was pH dependent, while in the case of Cu, it is related to the nutrient status of the growth medium of plants.