Acute toxicity of 30 pharmaceutically active compounds to freshwater planarians,Dugesia japonica

Pharmaceutically active compounds are produced worldwide and consumed in large quantities, so these chemicals are frequently detected in limnic environments posing potential ecological risks. Thirty pharmaceutically active compounds were selected for examination of their acute toxicity for freshwater planarians (Dugesia japonica). Among the 30 compounds tested, diclofenac, mefenamic acid, naproxen, propranolol HCl, and diphenhydramine HCl had a 48-h nominal LC₅₀ below10 mg L^?1, and for 18 chemicals, it exceeded 100 mg L^?1. The 96-h nominal LC₅₀ was below 10 mg L^?1 for diclofenac, mefenamic acid, naproxen, propranolol HCl, diltiazem HCl, diphenhydramine HCl, hydroxyzine HCl, and triprolidine HCl, and for 15 chemicals, it exceeded 100 mg L^?1. Among different therapeutic groups, analgesics were most toxic to planarians, and antibiotics were least toxic. Antihistamines and beta blockers varied in their acute toxicity to planarians. At the current environmental levels, none of the tested pharmaceutically active compounds may have acutely harmful impacts on aquatic invertebrates. To answer the question whether chronic, long-term exposure to pharmaceutically active compounds may entail ecological risks for aquatic ecosystems, further investigations with different end points in multiple species tests are needed.     

4种甲氧基丙烯酸酯类杀菌剂不同剂型对斑马鱼急性毒性效应

采用半静态法测定了4种甲氧基丙烯酸酯类杀菌剂原药及制剂对斑马鱼(Brachydonio rerio)急性毒性。结果表明,以实测浓度计,250 g·L~(-1)嘧菌酯悬浮剂对斑马鱼的96 h半数致死浓度(LC50)值为0.539 mg·L~(-1),急性毒性为高毒,95%嘧菌酯原药和50%嘧菌酯水分散粒剂对斑马鱼的LC50(96 h)值分别为1.09和1.21 mg·L~(-1),急性毒性均为中毒;98%啶氧菌酯原药和22.5%的啶氧菌酯悬浮剂对斑马鱼的LC50(96 h)值分别为0.0974和0.0972 mg·L~(-1),急性毒性均为剧毒;95%吡唑醚菌酯原药、15%吡唑醚菌酯悬浮剂和250 g·L~(-1)吡唑醚菌酯乳油对斑马鱼的LC50(96 h)值为0.0613、0.0549和0.0487 mg·L~(-1),急性毒性均为剧毒;95%醚菌酯原药和50%醚菌酯水分散粒剂对斑马鱼的LC50(96 h)值分别为0.468和0.702 mg·L~(-1),急性毒性均为高毒。这4种甲氧基丙烯酸酯类杀菌剂对斑马鱼均有较高风险,会对其他水生生物也存在潜在的风险。     

Larvicidal potential of different solvent extracts and oleanolic acid derivative from Coccinia indica against vector mosquitoes

The mosquito larvicidal activities of different solvent extracts (chloroform, ethyl acetate, and methanol) from the leaves of Coccinia indica were studied. Among the extracts tested, the methanol extract showed the highest larvicidal activity against early fourth instar larvae of Culex quinquefasciatus (LC₅₀?=?142?mg?L^?1), Aedes aegypti (LC₅₀?=?158?mg?L^?1) and Anopheles stephensi (LC₅₀?=?161?mg?L^?1) after 24?h of exposure period. The lowest larval mortality was observed with the ethyl acetate extract with the LC₅₀ values of 927, 872, and 730?mg?L^?1 against Cx. quinquefasciatus, Ae. aegypti, and An. stephensi, respectively. The chloroform extract showed the moderate larvicidal activity against the mosquito species tested with the LC₅₀ values from 446 to 546?mg?L^?1. A new triterpene was isolated from the methanol extract using bio-assay guided separation and identified as an oleanolic acid derivative by NMR, IR and mass spectral studies. The compound showed prominent larvicidal activity against Cx. quinquefasciatus, Ae. aegypti and An. stephensi with the LC₅₀ values of 5.6, 5.0, and 4.8?mg?L^?1, respectively. The results of the present investigation offer an opportunity for developing alternatives to rather expensive and environmentally hazardous organic insecticides.     

Oxidative stress in the freshwater cyprinid crucian carp (Carassius carassius L.) upon chronic exposure to endosulfan

Oxidative stress and antioxidant responses of crucian carp, upon chronic exposure to endosulfan, were evaluated in vivo. The lethal concentration (LC_50–96?h) was 70 μg L^?1; on its basis, the fish were exposed to endosulfan at 20, 35, and 50 μg L^?1 and autopsy was done on days 1, 2, 3, 4, 7, 14, 21, 28, and 35. Lipid peroxidation was induced in a concentration-dependent manner, being highest at 50 μg L^?1 (3/4 LC_{50–96 h}, sub-lethal concentration-I, SL-I) on day 4 (720% versus control), followed in its extent (490%) at 30 μg L^?1 (1/2 LC_{50–96 h}, sub-lethal concentration-II, SL-II) on day 7 and lowest (260%) at 10 μg L^?1 (1/4 LC_{50–96 h}, sub-lethal concentration-III, SL-III) on day 14. Glutathione showed a concentration- and time-dependent elevation in the initial phase, with highest level on day 4 (180%) at SL-I, but showed significant reduction in all test concentrations from day 21 of post-exposure. Superoxide dismutase was decreased significantly throughout the study, with highest reduction (63%) on day 4 at SL-I; catalase increased in all test concentrations up to day 14 but showed a significant decrease from the day 28 of post-exposure. The potential role of these parameters as indicators of pesticide pollution in aquatic systems is discussed.     

Toxicity of metals to tadpoles of the common Sunda toad,Duttaphrynus melanostictus

Tadpoles of the common freshwater Sunda toad, Duttaphrynus melanostictus (Amphibia, Bufonidae), were exposed for a 4-day period under laboratory conditions to copper (Cu), cadmium (Cd), zinc (Zn), lead (Pb), nickel (Ni), iron (Fe), aluminum (Al), and manganese (Mn) at various concentrations. Mortality was assessed and median times of death (LT₅₀) and lethal concentrations (LC₅₀) were calculated. LT₅₀ and LC₅₀ increased with the decrease in mean exposure times and concentrations for all metals. LC₅₀ (96?h) for Cu, Cd, Zn, Pb, Ni, Fe, Al, and Mn were 0.03, 0.3, 4.2, 1.5, 8.8, 0.4, 1.9, and 39?mg?L^?1, respectively. Cu was the most toxic to D. melanostictus, followed by Cd, Fe, Al, Pb, Zn, Ni, and Mn (Cu?>?Cd?>?Fe?>?Al?>?Pb?>?Zn?>?Ni?>?Mn). Duttaphrynus melanostictus is similarly sensitive to these metals as other amphibian tadpoles.     

Formation of resistance in the Chironomus plumosus to four pesticides over 45 generations

A study was conducted on the Chironomus plumosus larvae to determine initiation of resistance to four pesticides – chlorfenvinphos, chlorpyrifos, chlorpyrifos-methyl, and malathion. First generational LC₅₀ values were well within the threshold value for chironomids based upon the literature. Subsequent LC₅₀ values were observed to increase, indicating a lessening of the toxicity of the pesticides to the chironomid. In the case of chlorfenvinphos, the 96 h LC₅₀ for generations 1–23 was 6 µg L^?1, in generations 3–7 was 8.57 µg L^?1 and 11.14 µg L^?1 for generations 8–9. Generations 10–12 had an LC₅₀ value of 22.58 µg L^?1 and generation 13 had an LC₅₀ value of 35.08 µg L^?1. Generation 14 had an LC₅₀ value of 47.58 µg L^?1. Generations 15–19 and 20–24 had 96 h LC₅₀ values of 60.68 µg L^?1, 72.58 µg L^?1, 85.08 µg L^?1, 97.58 µg L^?1 and 110.08 µg L^?1, respectively. Generations 25, 26 to 30, 31 to 38 and 39 to 45 had 96 h LC₅₀ values of 160.42 µg L^?1, 210.7 µg L^?1, 262.24 µg L^?1 and 274.36 µg L^?1, respectively. The variation between LC₅₀ values was found to be statistically significant. This was observed for most pesticides tested. Larval size and life cycle duration was observed to change from generation to generation with the body size decreasing markedly from 1 to 0.3 cm with life cycle increasing from 7 to 39 days.     

Acute toxicity of ammonia to a freshwater teleost,Labeo bata larvae

Ammonia toxicity tests were performed with Labeo bata (bata) larvae of three different size groups. One hundred percent survival of larvae (500.0?±?4.0?mg) was recorded when exposed to ammonia concentrations of 1.0–13.56?mg?L^?1 at 96?h of exposure. Bata larvae exposed to ammonia concentrations of 15.8–25?mg?L^?1 showed 10–74% mortalities. The 96?h LC₅₀ value for 200 (±5), 250 (±2) and 500 (±4) mg bata larvae were 11.5, 16.8 and 22.5?mg?L^?1 un-ionised ammonia concentrations, respectively. When fish were exposed to different doses of ammonia, behavioural changes immediately occurred even at the lowest dose. At first, the fish became hyperexcitable, the skin darkened and they showed an increased ventilation frequency, fish behaviour became normal, 24?h after exposure. A 96?h LC₅₀ value of un-ionised ammonia showed direct relationship with the increasing size of bata larvae.     

Deltamethrin-induced alterations in serum calcium and prolactin cells of a freshwater teleost,Heteropneustes fossilis

Deltamethrin, a pyrethroid insecticide widely used in many countries, causes significant adverse effects in aquatic ecosystems. The concentration of deltamethrin in water reservoirs and the run off from agricultural areas (in water) in many countries range up to 24.0?µg?L^?1 which is higher than the recommendation of the European Union standard. Hence, in this study the effects of deltamethrin were investigated, i.e. its toxic impacts on the freshwater catfish Heteropneustes fossilis in terms of serum calcium and prolactin cells (located in the rostral pars distalis region of the pituitary). The fish were subjected to deltamethrin for a short-term experiment (96?h; 1.5?µg?L^?1 e.0.8 of 96?h LC₅₀) and a long-term experiment (28 days; 0.37?µg?L^?1 e.0.2 of 96?h LC₅₀). After short-term deltamethrin exposure, serum calcium levels decrease. No histological change in prolactin cells is noticed throughout the short-term experiment. Fish exposed to deltamethrin for 7 days also exhibit a decrease in serum calcium level. This decrease persisted until the end of the experiment (28 days). Prolactin cells of fish treated for 14 days with deltamethrin exhibit increased nuclear volume and degranulation, increasing progressively from the 21st day onwards. After 28 days, a few degenerated cells are discerned. The results of this study show that deltamethrin is moderately toxic for the freshwater fish H. fossilis by producing adverse effects on serum calcium and prolactin cells. Hence, it should be used with caution in areas near fish-inhabited waters.     

A comparison of two methods to observe the effects of dual metal exposure to the chironomus larvae

A study was conducted to investigate the effects of metal mixtures on the Chironomus plumosus. Two methods of preparing the mixtures were used, the Toxic Unit (TU) method and the parts per million (PPM) + PPM method. A comparison was undertaken between the two methods to observe the ease with which (1) experiment could be conducted and (2) whether LC₅₀ values could be obtained. In this study, it was noted that the PPM + PPM method was an easier method to conduct metal mixture experiments as the LC₅₀ values could be easily calculated for individual metals and compared with the LC₅₀ values of metals obtained from single metal experiments. The findings showed that the LC₅₀ value for zinc (Zn) was more toxic for chromium (Cr) when it was reduced from 9.6 to 0.3 mg L^?1, with silver the LC₅₀ value became 3.6 mg L^?1, 4.5 mg L^?1 with nickel (Ni). In the PPM + PPM method, the LC₅₀ value for Zn was observed to be 10.2 mg L^?1 when in mixture with chromium, 23.8 mg L^?1 with silver, and 13.4 mg L^?1 with Ni. Changes in the 96 h LC₅₀ value were found to be significant for all metals.     

Toxic effects of phosphoramidate on Paramecium sp. with special emphasis on respiratory metabolism,growth, and generation time

The toxicological impacts of the increasing number of synthetic compounds present in the aquatic environment are assessed predominantly in laboratory studies where test organisms are exposed to a range of concentrations of single compounds. Protozoan cells are often used as bioindicators for the presence of xenobiotic compounds. In this article, we describe the inhibitory effect of a synthetic phosphoramidate derivative at different concentrations (40, 60, and 80?µmol?L^?1) on Paramecium sp., affecting its growth (proliferation) in concentration-dependent manner, as well as the generation time and response percentage. The LC₅₀ value determined for these protozoa was estimated at 60?µmol?L^?1 on 24?h of exposure. The respiratory metabolism of protozoan is perturbed at three concentrations, noting that the oxygen consumption was significantly increased at high concentrations after 18?h of exposure. In addition, the data can be used as reference values in further testing with other pesticides.     

Uptake and toxicity of some pesticides on three freshwater fishes (Oreochromis niloticus,Clarias gariepinus and Chrysicthys nigrodigitatus)

Studies were carried out to determine the toxicity of some selected pesticides on fresh water fish in a tropical environment. The uptake of the pesticides lindane, pentachlorophenol (PCP), and propoxur, which are frequently used by farmers and industrialists were studied in concrete ponds at the University of Cape Coast, in Ghana. The fishes used for the study were Oreochromis niloticus, Clarias gariepinus, and Chrysicthys nigrodigitatus, and were obtained from cultured ponds in the Cape Coast District and Mankessim in the Central Region and Weija Dam, in the Greater Accra region of Ghana. Single high lethal concentration (SD) or acute treatment and cumulative/chronic (or multiple minor) lethal concentration (CD) treatment were employed in administering the pesticides to the fishes via water. Gas chromatograph (GC) Electron Capture Detector (ECD) analysis was done on the dead fishes to see the extent of ingestion. The LC₅₀ values obtained for lindane on the three fish samples were as follows: Chrysicthys?–?0.38?mg?L^?1; Oreochromis?–?0.42?mg?L^?1, and Clarias?–?1.2?mg?L^?1. Mortalities occurred in fish within 3–5 days of application. For the PCP on Chrysicthys, Oreochromis, and Clarias species the LC₅₀ values were 0.42, 0.32 and 0.64?mg?L^?1, respectively for over a 2–3 day period. For a three-time influx period of propoxur the LC₅₀ for Chrysicthys, Oreochromis, and Clarias, were 22.0, 30.40, and 45.04 (all in mg?L^?1), respectively. The results obtained indicated that the pesticides had adverse effects on the general growth and reproduction of fishes. Gonadosomatic indices also showed that the pesticides affected the development of the body, the gonads, and their reproduction.     

In vitro kinetic characterization of inhibition of acetylcholinesterase by organophosphate and carbamate compounds in food animals

Organophosphate, dichlorvos (2,2-dichlorovinyldimethylphosphate, DDVP), and diazinon (DZN) as well as carbamates are currently found in contaminated water, dust, soil, agricultural waste, and elsewhere in the environment, and can be harmful after accidental or deliberate exposure. Intoxication by these compounds causes a generalized cholinergic crisis due to the inhibition of acetylcholinesterase (AChE), whose major physiological role in mammalian tissues is in nervous transmission. The half-maximal inhibitory concentration (IC₅₀) was found to be higher for carbaryl than DDVP and DZN, about 3–4 times in liver and about 4–6 times in muscles. The half time (t _1/2) of inhibition (8?µmol?L^?1 DDVP or DZN; 40?µmol?L^?1 carbaryl) ranged between 4 and 16?min and decreased as follows: pig?>?cattle?>?sheep for liver tissue, and cattle?>?sheep?>?pig for muscle. Rate constants of inhibition (k _i) ranged between 43?×?10^?3 and 168?×?10^?3?min^?1 for liver and from 46?×?10^?3 to 115?×?10^?3?min^?1 for muscles. Very little residual AChE activity (<6.1%) was seen in liver, but more was seen in muscle (<17%).     

Low uptakes of Cd,Cu, and Zn in Dugesia japonica,a freshwater planarian with higher tolerance to metals

Although a freshwater planarian is proposed as a potential model for studying toxicities of environmental pollutants, they are less sensitive to metal exposures, compared to other freshwater invertebrates. We hypothesised that the metal uptake is low in treated planarians, which may lead to their higher tolerance to metals. The acute toxicities of Cu and Zn to Dugesia japonica were determined. The 24- and 48-h median lethal concentrations were 4.50 and 3.46?mg?L^?1, respectively, for Cu, and 41.97 and 37.79?mg?L^?1, respectively, for Zn. Accordingly, combining with the data on Cd in our previous study, higher tolerance of planarians to metals was revealed. Also, tissue levels of Cd, Cu, and Zn in treated D. japonica were determined, and their dissolved uptake rate constants (k₁) were calculated. Results showed that bioaccumulations of these metals in D. japonica were, indeed, lower than that in other freshwater invertebrates having higher susceptibilities to these metals.     

The influence of solid matrices on arsenic toxicity to Corophium orientale

In marine ecosystems, benthic organisms are really important because they are the first step in the transfer of contaminants from environment to biota. To this end, this study focused on biological assays with the amphipod Corophium orientale exposed to two different molecules of arsenic: arsenate (As^V), the most abundant form in sediments, and dimethyl-arsinate (DMA), expected to be moderately toxic as an intermediate in the process of detoxification. The toxicity of arsenic compounds was measured after exposure to three different matrices: water, spiked natural sediment and inert spiked quartz sand. LC₅₀ values were calculated for each exposure, and the results confirmed the highest toxicity of As^V, in addition to underlining the importance of matrix of exposure. Water exposure was the matrix which presented the highest toxicity for inorganic arsenic (As^V LC₅₀=3.51 mg L^?1 vs DMA LC₅₀=54.65 mg L^?1), spiked natural sediment demonstrated its capability to chelate arsenate toxicity (As^V LC₅₀=34.27 mg kg^?1 vs. DMA LC₅₀=52.19 mg kg^?1) and spiked quartz sand presented intermediate values for As^V (LC₅₀=25.26 mg kg^?1), whereas for DMA a lower toxicity was registered (LC₅₀=872.35 mg kg^?1). This study can provide some useful data linked with chemical speciation of arsenic and exposure matrix, for improving the correct management of contaminated sediment.     

Acute and chronic toxic effect of lead (Pb) and zinc (Zn) on biomarker response in post larvae of Penaeus monodon (Fabricus, 1798)

Metal pollution produces damage to marine organisms at the cellular level possibly leading to ecological imbalance. The present investigation focused on the acute and chronic toxicity of lead (Pb) and zinc (Zn) by examining the effects of biomarker enzymes in post-larvae of Penaeus monodon (Tiger prawn). Antioxidant biomarker responses such as lipid peroxidation (LPO) and catalase (CAT) activity for Pb and Zn were determined following chronic exposure. Acute Lethal Concentration ₅₀ (LC₅₀) values observed in the study at 96?h for Pb and Zn at 5.77?±?0.32?mg?L^?1 and 3.02?±?0.82?mg?L^?1, respectively. The estimated No Observed Effect Concentration and Lowest Observed Effect Concentration values for Pb were 0.014 and 0.029?mg?L^?1 and that recorded for Zn was 0.011 and 0.022?mg?L^?1, respectively. Among the two metals studied, toxicity of Zn was found to be greater to P. monodon than Pb. The activities of antioxidant defense enzymes and total protein content differed significantly from control following exposure to both metals. Overall, the biomarker studies demonstrated that alterations in antioxidant enzymes and induction of LPO reflect the consequences of heavy metal exposure in P. monodon.     

Acute toxicity of industrial endocrine-disrupting chemicals,natural and synthetic sex hormones to the freshwater planarian,Dugesia japonica

Many chemicals commonly occurring in surface water are found to be hormonally active. Fourteen compounds including four sex hormones, three synthetic hormones, one non-steroidal anti-androgen, and six industrial endocrine-disrupting chemicals were selected to examine their acute toxicities on the freshwater planarian, Dugesia japonica. Among all test chemicals, diethylstilbestrol had the highest toxicity. Relatively, the toxicities of the synthetic hormones were higher than the natural ones. Among the six industrial endocrine-disrupting chemicals tested, bisphenol A was the most toxic, with a 48-h LC₅₀ of 8.3 mg L^?1. Overall, the natural or synthetic hormones, other than estrone and estriol, were more acutely toxic to planarians than the industrial endocrine disruptors tested. The acute toxicities of the test chemicals to Dugesia japonica were in the same order of magnitude as those to Daphnia magna based on available published data. Despite this study used non-environmentally relevant levels, such kinds of chemicals do not exist singly in the aquatic environment, but typically act in synergistic and/or additive ways in the complex environmental mixtures, and the total estrogen equivalents are much higher than a single chemical. This study stands as a starting point for other acute and chronic assays, namely using mixtures.     

Prevalence of organochlorine pesticide residues in groundwaters of Kasargod District,India

The Kasargod District is the northernmost district of Kerala State and is bound between the north latitudes 12°02′27″ and 12°47′35″ and east longitudes 74°51′54″ and 75°25′25″. The present study highlights the contamination levels of organochlorine pesticides in open wells of the Kasargod District from 2010 to 2011. Maximum contamination of organochlorine pesticides (OCP's) was observed for endosulfan followed by hexachlorobenzene (BHC). Contamination levels of α-endosulfan were higher at Panathur (58?µg?L^?1) and next to Periya (37?µg?L^?1) in the postmonsoon season of 2010. During premonsoon 2011, the residue levels of α-endosulfan were higher at Panathady (56?µg?L^?1) followed by Rajapuram (40?µg?L^?1). Contamination levels of the BHC isomers exhibit the order of γ-BHC?>?α-BHC?>?β‐BHC, showing that γ-BHC represents 62% of the total OCP residues in premonsoon 2010. Among the studied OCP's, concentration levels of DDT were below detection limit. Maximum concentrations of the sum of all OCP residues were observed at Mulleria and Cheemeni (premonsoon 2010), followed by Panathur and Periya (postmonsoon 2010), and lowest in Panathady and Rajapuram (premonsoon 2011). Residues of OCP's were lowest in Paettikundu and Cheravatur in all the three seasons.     

Quantal response of Paleomonetes africanus in locally formulated drilling mud lubricants

The toxic potency of three locally formulated drilling mud lubricants against Paleomonetes africanus, a brackish water crustacean commonly found in the oil producing Niger Delta (Nigeria), was evaluated. A 96?h exposure to quantal (lethal) doses of the mud lubricants gave LC₅₀ (median lethal concentration) of 350, 620 and 210?mg?L^?1, respectively. The mud lubricant with the highest molecular weight of 40,000?g?mol^?1 (lubricant 2) had the least toxicity on the organisms, attributable to the reduced rate of diffusion into the vital cells of the test organism. The higher toxicity of lubricant 3 over lubricant 1, despite its higher molecular weight, might be as a result of its higher acid value of 21.60?mg?KOH?g^?1. The results show that the test lubricants would be environmentally friendly within the baseline concentrations and lethal times (LT₅₀) measured.     

Differential effects of UV-B radiation fluence rates on growth,photosynthesis, and phosphate metabolism in two cyanobacteria under copper toxicity

This work was undertaken to ascertain the impact of different fluence rates of ultraviolet-B (UV-B) radiation on two cyanobacterial biofertilizers, Phormidium foveolarum and Nostoc muscorum, growing under copper toxicity. Copper (2 and 5?µmol?L^?1) and high UV-B fluence rate (UV-B_H; 1.0?µmol?m^?2?s^?1) decreased the growth, pigment content, photosynthetic oxygen yield, phosphate uptake, and acid phosphatase activity in both the strains analyzed after 24 and 72?h of experiments, and combined exposure further enhanced the toxic effects. Respiration and alkaline phosphatase activities were stimulated appreciably. The damaging effect was shown on the order on pigments: phycocyanin?>?chlorophyll a?>?carotenoids, and on photosystems: whole chain photosynthetic reaction?>?photosystem II?>?photosystem I. Partial recovery in the photosystem II activity in the presence of artificial electron donors; diphenyl carbazide (DPC), hydroxylamine (NH₂OH), and manganese chloride (MnCl₂) pointed out the interruption of electron flow on the oxidation side of photosystem II. Unlike UV-B_H, low UV-B fluence rate (UV-B_L; 0.1?µmol?m^?2?s^?1), rather than causing damaging effect partially, alleviated the toxic effects of Cu. This study suggests that the cyanobacterium P. foveolarum is less sensitive against UV-B_H and excess Cu (2 and 5?µmol?L^?1), thus P. foveolarum may be used as a biofertilizer for sustainable agriculture.     

Cardioprotective effect of Lagenaria siceraria (Mol) ameliorates isoproterenol-induced cardiac toxicity in rats by stabilizing cardiac mitochondrial and lysosomal enzymes

A study was conducted to determine the effect of the chironomid larval tube on heavy metal toxicity. Silica gel was provided as a substrate with which the chironomid larvae could construct larval tubes. A series of 96 h LC₅₀ bioassays were conducted with and without the presence of the silica gel substrate, to allow comparison between the two test conditions. From the study it was observed that a 10–15% variation in LC₅₀ value occurred. The 96 h LC₅₀ for copper at the temperatures 35, 28, 20, 15, and 10°C was observed to be 7.5, 9.5, 7.5, 4.5, 3.6, and 2.8 mg L^?1, respectively. The same test when conducted in the presence of silica gel produced LC₅₀ values of 26.50, 33.50, 15.82, 12.77, and 9.94 mg L^?1 for the same temperatures. This would demonstrate that the larval tube did, to some extent, protect the larvae from the effects of the heavy metal.