A novel extraction method for analysing available sulfamethoxazole in soil

A novel extraction method was established to determine the water-extractable (available) content of sulfamethoxazole (SMX) in soil. The SMX imprinted polymers (MIPs) were synthesised and the performance was evaluated by Fourier transform infrared spectroscopy, scanning electron microscopy and binding experiments. Results showed that the MIPs exhibited good selectivity for SMX, so the MIPs were applied as a sorbent. SMX in soil was extracted by water, sorbed from the extract to MIPs and analysed with a high performance liquid chromatography (HPLC) after its desorption from MIPs. Meanwhile, the classic organic solvent extraction was employed to measure the total SMX content in soil. Results showed that when SMX level in spiked soils varying from 1.0–500?μg?kg^?1, the observed recoveries of available SMX contents ranged from 63.27?±?3.11% to 82.11?±?2.77% (n?=?3), while the total SMX varied between 89.59?±?1.65% and 97.64?±?3.92% (n?=?3). The detection limit of the developed method for SMX in soils was 0.05?μg?kg^?1. Available SMX contents in five field soil samples ranged from 0.13 to 4.14?μg?kg^?1, which were only 0.35–25.40% of the respective total SMX contents. Results from this study manifest the importance of the extents of SMX immobilisation with different soils for assessing SMX's ecological and human health risks.     

Development of a complex-based flow injection spectrophotometric method for determination of the herbicide pinoxaden in environmental samples

A flow injection (FI) spectrophotometric method for the determination of the herbicide pinoxaden (PXD) has been proposed. PXD was converted in alkaline media with hydroxylamine hydrochloride to the hydroxamate salt. The salt was reacted with iron(III) chloride, and the absorbance of the red colored tris iron hydroxamate complex was measured at 500?nm using a FI system. The method was found to be linear between 0.5 and 40?mg?L^?1 with a molar absorptivity of 1.53?×?10^4?L?mol^?1?cm^?1. The limit of detection and limit of quantification were found to be 0.1?±?0.01?mg?L^?1 and 0.6?±?0.05?mg?L^?1, respectively. Any interference of fenoxaprop-p-ethyl (FE) was avoided by the separation of PXD by liquid chromatography with a mixture of dichloromethane and n-hexane (1?:?1) as eluent. The method was applied to the determination of PXD in soil, water, and wheat grains with percent recoveries of 98?±?2, 100?±?2, and 98?±?5, respectively. Sample throughput of 60 samples per hour was achieved under optimized conditions.     

Biological effects of water soluble fraction of crude oil on the Arctic sea ice amphipod Gammarus wilkitzkii

With increasing petroleum related activity in the Barents Sea and the subsequent risk of accidents, there is a demand for knowledge about the effect of oil pollution in Arctic ecosystems. In the present study, the sea ice amphipod Gammarus wilkitzkii was exposed to a water soluble fraction (WSF) of oil in two experiments, using a rock column system. First, three groups of adult females were exposed for 36 days for control (n=9), low dose (n=10) (initial and final oil concentration: 14, 115 and 120 μ g/l) and high dose (n=10) (initial and final oil concentration: 78, 764 and 395 μ g/l). Total oxyradical scavenging capacity (TOSC), malondialdehyde (MDA), catalase activity (CAT), respiration rate and mortality were measured. In the second experiment, moulting rate was measured in immature individuals of two groups, control (n=10) and exposed (n = 20), over 113 days. No mortality was observed in either of the experiments. There was a dose-related significant increase in respiration rate (40.69 (±22.82), 55.63 (±20.98), 94.57 (±22.80)) mg O₂ h^?1g ww^?1 in control, low dose and high dose, respectively. A higher MDA level was detected in the low dose group (25.04 (±6.00) nmol g^?1 of tissue) compared to both control (20.44 (±2.62) nmol g^?1 of tissue) and high dose groups (20.93 (±4.79) nmol g^?1 of tissue). Likewise, the low dose group had the highest value of TOSC towards hydroxyl (727.74 (±475.58), 1157.58 (±278.62), 1067.30 (±369.22) TOSC unit value mg^?1 of protein for control, low and high dose, respectively). Although no difference in the catalase activity between control and exposed groups was detected, higher activity was measured at 0 °C (average: 248 μ mol^?1 min^?1 mg^?1 of protein) than at the standard temperature for catalase measurement of 25 °C (average: 140 μ mol^?1 min^?1 mg^?1 of protein), indicating a need to optimise the standard operational procedure when working with Arctic organisms. No effect of WSF exposure on moulting rate was detected. In conclusion, little mortality was observed during the experiments, indicating some degree of tolerance levels. However, the biomarker results indicated sub-lethal effects in G. wilkitzkii after exposure to WSF.     

Investigation of naphthalene contamination in olive oil from Greece

Abstract

Incidents of using naphthalene in olive orchards as a repellent of olive fly (Bactrocera oleae) have recently been recorded. Naphthalene, the primary ingredient of mothballs, is described to be diluted in plastic bottles and hanged on the trees releasing its characteristic odor which is supposed to repel olive fly. Since naphthalene has been characterized as possibly carcinogenic to humans and animals, it was considered essential to monitor olive oils regarding their potential contamination with this specific pollutant. To this goal, a simple, sensitive and reliable analytical method for the determination of naphthalene in olive oil has been developed. The method involves extraction with ethyl acetate: acetonitrile (1:1) and determination of naphthalene using gas chromatography triple quadrupole mass spectrometry. The method was validated in four fortification levels, 0.005, 0.01, 0.05, and 0.1?mg/kg in accordance with the EU requirements. The obtained results were acceptable as far as validation criteria are concerned, given recoveries between 71% and 87% and RSDs between 5% and 24%. The limit of determination for naphthalene was set at 0.005?mg/kg based on the lowest concentration level being validated with acceptable accuracy. The analytical technique was successfully applied to 75 olive oil samples collected from oil mills in Greece and no positive results of naphthalene were detected.     

Concentrations of lead in cosmetics commonly used in South Africa

In this study, we present evidence of lead in lip liner, oil absorbent powder, mascara, concealer, lipsticks, lip gloss, and foundation. The samples were analyzed for lead by flame atomic absorption spectrometry. The levels of lead in concealer, mascara, lip liner, and oil absorbent powder were found to be 7.4?±?1.3?µg?g^?1, 15.8?±?0.2?µg?g^?1, 29.0?±?9.2?µg?g^?1, and 17.3?±?2.9?µg?g^?1, respectively. The levels of lead in lipsticks, lip gloss, and foundation ranged from not detected to 73.1?±?5.2?µg?g^?1, 4.7 to 11.7?±?2.8?µg?g^?1, and 7.8 to 32.9?±?1.4?µg?g^?1, respectively. Thus, in the majority of samples, the concentrations of lead are higher than the USFDA maximum permissible concentration of 0.10?µg?g^?1 in candy and cosmetics. In lipsticks alone, only 25% of 40 samples were found to meet the requirement. Therefore, continuous use of cosmetic products in which lead concentration exceeds the maximum permissible level may pose a health hazard for the female population of South Africa in the long run.     

Toxicokinetics,metabolism, and microsomal studies of chlorpropham in rats

A study on the toxicokinetic behavior, metabolism of chlorpropham, and its effect on cytochrome P₄₅₀ from liver microsomes was carried out in albino rats after a single and consecutive oral administration at 500?mg?kg^?1 body weight for 10 and 20 days. Chlorpropham was detected in the blood at 0.08?h (11.43?±?1.72?µg?mL^?1) reaching a maximum concentration at 2?h (30.90?±?2.55?µg?mL^?1) and a minimum at 48?h (1.95?±?0.20?µg?mL^?1) after a single oral administration of 500?mg?kg^?1. The absorption rate constant (K _a) was 0.66?±?0.48?h^?1. The Vd _area (18.01?±?2.78?L?kg^?1) and t _1/2 β (12.23?±?1.96?h) values suggested a wide distribution and long persistence of the compound in the body, respectively. The higher Cl_R (0.82?±?0.00?L?kg^?1?h^?1) compared to Cl_H (0.18?±?0.02?L?kg^?1?h^?1) value indicated that a major portion of chlorpropham was excreted through the urine (30%) compared to the faeces (2.81%). Chlorpropham residue was detected in all tissues of rat at 0.25?h while its metabolite, meta-chloroaniline was detected in liver, kidney, heart, lung, and spleen tissue at 0.25?h. Meta-chloroaniline was not detected in skeletal muscle, brain, fat, and stomach tissue at any time of the observation period. Maximum concentrations of chlorpropham and meta-chloroaniline were detected at 2?h (except in the spleen), and minimum concentrations of chlorpropham at 24 (heart, lung, spleen, skeletal muscle, and stomach) and 48?h (liver, kidney, brain, and fat tissue) respectively; and meta-chloroaniline at 24?h (except heart and spleen). The tissue half-life of chlorpropham in rat varied from 3.80 to 11.60?h. Repeated oral administration of chlorpropham at 500?mg?kg^?1 for 10 and 20 days caused an induction of the liver microsomal pellet of rat.     

The potential of Berkheya coddii for phytoextraction of nickel,platinum, and palladium contaminated sites

The study involved assessing the potential of the native plant species (Berkheya coddii) for the phytoextraction of nickel, palladium, and platinum contaminated sites. Plant and soil samples were randomly collected from Barberton area, near Agnes mine, Mpumalanga Province, South Africa. Samples were analysed for total nickel, palladium, and platinum concentrations together with other elements found in the soil and in the plants' roots, and leaves. Soil versus leaves and soil versus roots uptake of these metals by the plant were compared. The mean concentration of nickel in the leaves/canopy was found to be 13,980?±?10,780?mg?kg^?1?dry mass, in the roots it was 2046?±?789?mg?kg^?1 dry mass, and in the soil it was 1040?±?686?mg?kg^?1?dry mass. This resulted in a mean concentration ratio in the leaves to soil of 13.44. The platinum mean concentration in the leaves was 0.22?±?0.15?mg?kg^?1?dry mass, in the roots it was 0.14?±?0.04?mg?kg^?1?dry mass, and in the soil it was 0.04?±?0.03?mg?kg^?1?dry mass. This resulted in a mean concentration ratio in the leaves to soil of 5.5. Palladium was found to have a mean soil concentration of 0.07?±?0.045?mg?kg^?1?dry mass. The mean concentrations in the roots and in the leaves were 0.18?±?0.07 dry mass and 0.71?±?0.52?mg?kg^?1?dry mass, respectively. This gave a mean concentration ratio in the leaves to soil of 10.1 for palladium. Other elements that were found to have a mean concentration ratio in the leaves to soil of around 2.5 or above are sodium, potassium, magnesium, calcium, and sulfur. Berkheya coddii was found to be most efficient in accumulating nickel, palladium, and platinum from the soil. The results for the first time revealed that the plant may have the potential to uptake platinum and palladium; both metals are in the same group of the periodic table as nickel.     

Larvicidal activity of neem and karanja oil cakes against mosquito vectors, Culex quinquefasciatus (say), Aedes aegypti (L.) and Anopheles stephensi (L.)

Larvicidal effect of neem (Azadirachta indica) and karanja (Pongamia glabra) oil cakes (individuals and combination) was studied against mosquito species. Both the oil cakes showed larvicidal activity against the mosquito species tested. The combination of neem and karanja oil cakes in equal proportion proved to have better effect than the individual treatments. The combination of the two oil cakes recorded an LC95 of 0.93, 0.54 and 0.77% against the mosquitoes, Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi respectively The increase in efficacy of the combination treatment over individuals in all the mosquito larvae tested was found to range about 4 to 10 fold in terms of LC50 and 2 to 6 fold in terms of LC95.     

Morpho-histological and enzymatic alterations in earthworms Drawida willsi and Lampito mauritii exposed to urea,phosphogypsum and paper mill sludge

The effects of varying concentrations of urea, phosphogypsum and paper mill sludge (PMS) on the morphology, histology, tissue protein content, lipid peroxidation (LPX), activities of lactate dehydrogenase (LDH), acetylcholinesterase (AChE) and catalase in earthworms Drawida willsi and Lampito mauritii have been studied over an exposure period of 24?hr. Integumentary lesions, clitellar swelling and loss of pigmentations were found to be major morpho-pathological changes in the worms. Histology indicated cuticular damage, ruptured epithelium and muscle fibres with accumulation of cellular debris. Lowest tissue protein content (57.02?±?4.02?mg/g tissue) and highest LPX (0.113?±?0.04, 0.137?±?0.08?nmol/mg protein) were noticed in D. willsi at a high concentration of PMS, whereas highest tissue protein content was observed in L. mauritii (115.32?±?7.18?mg/g tissue) with the same treatment. In both the species, LDH activity was minimum at a high concentration of urea (0.172?±?0.02; 0.247?±?0.08?U/mg protein). AChE activity was highest (0.099?±?0.002?U/mg protein) at a high concentration of PMS in D. willsi, whereas catalase activity was the maximum (0.338?±?0.02?U/mg protein) at high concentrations of PMS in L. mauritii. The study indicated that morpho-histological and enzymatic alterations in these earthworms exposed to agrochemicals could be useful biomarkers to evaluate soil toxicity.     

Lead and copper in the sclerotium of the mushroom Pleurotus tuber-regium (Osu): assessment of contribution to dietary intake in southeastern Nigeria

The concentrations of lead and copper in sclerotium of the mushroom Pleurotus tuber-regium widely consumed in Southeastern Nigeria were determined. The specimens purchased from different markets were mineralized with H₂SO₄ and H₂O₂ and analyzed using flame atomic absorption spectrophotometer. The concentrations of Pb ranged from 0.2?±?0.1?mg?kg^?1 to 0.8?±?0.5?mg?kg^?1 with approximately 91% of the results being below 1?mg?kg^?1. The concentrations of Cu ranged from 0.5?±?0.2?mg?kg^?1 to 1.2?±?0.6?mg?kg^?1 with 78% of the results below 1?mg?kg^?1. The results were compared with the literature and levels set by regulatory authorities, with the conclusion that the consumption of sclerotium does not pose a toxicological risk. The low Pb content of the studied products would contribute to only about 1% of the provisional tolerable weekly intake of Pb. The Cu contents would contribute to nutritional intake of the metal in the general population. It is recommended that the outer layers of the sclerotia be properly scrapped before use to reduce metal contamination from exogenous sources.     

Assessment of toxicity of two types of drill cuttings from a drilling rig on the Trinidad East coast using Metamysidopsis insularis

This study investigated the relative toxicity of water-based cuttings (WBC) and synthetic oil-based cuttings (SOBC) to the marine species, Metamysidopsis insularis. Results obtained indicate that SOBC (LC₅₀ 1.2 (0.85–1.6)%) was more toxic to M. insularis than WBC (LC₅₀ 9.9 (8.3–11.8)%), with similar metal contents in both types of cuttings. The elevated levels of metals found in the cuttings when compared to surficial sediments may be due to both drilling fluids, as well as the rock strata from which the cuttings were obtained. Furthermore, total petroleum hydrocarbon (TPH) analyses demonstrated significantly higher concentrations of TPH present in SOBC (14,680?±?1250?mg?kg^?1) compared to WBC (860?±?115?mg?kg^?1). This may also be due to the increased depth and hence oil bearing rock formations in the selected sampling area, along with the associated synthetic oil-based drilling fluid. These findings therefore supply evidence that drill cuttings after treatment prior to discharge are potentially toxic to marine organisms.     

Manganese toxicity in Drosophila melanogaster: extension of the life span by resveratrol

The effect of resveratrol was studied on the life span and motor activity of Drosophila melanogaster treated with manganese (Mn). Two days after emerging from the pupa, male wild type D. melanogaster were fed for 13 days with corn media containing 15?mM Mn. Thereafter, Drosophila were divided into six groups of 300 flies each: (1) the flies remained treated with Mn; (2) began treatment with 0.43?mM resveratrol (Mn-resveratrol group); (3) received no additional treatment (Mn-no treatment group); (4) simultaneously fed with Mn and resveratrol (Mn?+?resveratrol group). In addition, a control (5) with no treatment and another group (6) fed only with resveratrol after emerging from the pupa were included. All Mn-treated flies (group 1) were dead on the 25th day. The life span in the resveratrol group was 91?±?0.33 days (mean?±?S.E.M.) and in Mn-resveratrol flies was 83?±?2 days. These two values were significantly higher than those detected in the control (5) and Mn-no treatment (group 3) flies whose life span were 68?±?0.33 and 67?±?2.31 days, respectively. The Mn?+?resveratrol-fed flies had a markedly higher life span (31?±?1.53 days) than Mn-fed flies (23?±?0.88 days). In the flies that received Mn (Mn and Mn?+?resveratrol groups), the motor activity decreased significantly with respect to control (groups 5) and the Mn–resveratrol and resveratrol groups. In conclusion, resveratrol increased significantly the life span of Mn-treated D. melanogaster.     

Ultrasound-assisted emulsification/microextraction based on solidification of trace amounts of thallium prior to graphite furnace atomic absorption spectrometry determination

In the present work, determination of ultratrace amounts of thallium in water samples was performed by ultrasound-assisted emulsification microextraction based on solidification of a floating organic drop as sample preparation method prior to furnace atomic absorption spectrometry. 1-(2-Pyridylazo)-2-naphthol was used as chelating agent. The factors influencing the complex formation and extraction, such as pH of the aqueous solution, the type and the volume of extraction solvent, the volume of chelating agent solution, and the extraction time were investigated. Under optimized conditions, the enrichment factor was 200. The calibration graph was linear from 0.2 to 10.0 μg L^?1 with a correlation coefficient of 0.9966, the detection limit was 0.03 μg L^?1 and reproducibility was ±3.3% (C = 5.0 μg L^?1, n = 8). The method was successfully applied for the determination of thallium in water samples.     

Efficiency of Zn phytoextraction,biomass yield and formation of low-molecular-weight organic acids in S × rubens – a hydroponic experiment

The aim of the study was to estimate Zn phytoextraction and changes in biomass of S?×?rubens growing in modified Knop's solution with different levels of Zn addition (0.5, 1.0, 2.5 and 5.0?mM). Obtained results were correlated with secretion of selected low-molecular-weight organic acids (LMWOAs) in the rhizosphere, roots and leaves. An increase in Zn concentration in Knop's solution resulted in Zn accumulation in roots, shoots and leaves. The highest accumulation was observed for plants growing in 5?mM Zn, at concentration levels 4741.36?±?98.66, 1227.31?±?16.57 and 2241.65?±?34.90?mg?kg^?1 DW in roots, shoots and leaves, respectively. The bioaccumulation factor and the translocation factor for plants growing in 0.5, 1.0 and 2.5?mM Zn clearly indicate that this Salix taxon is an effective Zn accumulator. The general reduction of Salix biomass with an increase in Zn concentration in the solution was observed. In the rhizosphere, the total LMWOA concentration was almost 0.93?µmol?kg^?1 DM for control (Zn free) plants, while for 5.0?mM of Zn it was 4.9?µmol?kg^?1 DM. Increasing concentrations of acids were observed in roots (1.34 for the control and 5.57?µmol?kg^?1 DM for plants treated with 2.5?mM of Zn).     

Ligand-less in situ surfactant-based solid phase extraction for preconcentration of silver from natural water samples prior to its determination by atomic absorption spectroscopy

A simple and rapid ligand-less in situ surfactant-based solid phase extraction method for preconcentration of silver from water samples is developed. In this method, a cationic surfactant containing a proper alkyl group (n-dodecyltrimethylammonium bromide) is dissolved in the aqueous sample and then a proper ion-pairing agent (ClO₄^?) is added. Due to the interaction between surfactant and ion-pairing agent, solid particles are formed and used for adsorption of silver carbonate. After centrifugation, the sediment is dissolved in 2.0 mL 1 M HNO₃ in ethanol and then aspirated directly into the flame atomic absorption spectrometer. Variables affecting the extraction efficiencies such as pH, concentrations of surfactant and CO₃^2?, ion pair concentration, and extraction time, are optimized. Under such conditions, the calibration curve is linear from 3 to 700 μg L^?1. Detection limit is 1.1 μg L^?1 with an enrichment factor of 37. The relative standard deviation for eight replicate measurements of 100 μg L^?1 is 2.1%. The method has been applied for the determination of silver in water samples.     

Screening of endogenous antioxidants in some medicinal plants

The evaluation of antioxidant enzymes and antioxidant metabolites was done in leaf tissues of Azadirachta indica, Butea monosperma, Cassia fistula, Mangifera indica, and Syzygium cumini growing in the Thar Desert, Rajasthan, India. The plants are naturally exposed to drought stress and high temperatures during summer. Enzymatic reactive oxygen species (ROS) scavenging mechanisms in plants include superoxide dismutase (SOD), ascorbate peroxidase (APX), glutathione reductase, catalase, and guaiacol peroxidase, and non-enzymatic antioxidants including the carotenoids, proline, and vitamin C were studied. The strategies to cope up with ROS under these extreme conditions are plant-specific. The highest activity of APX was found in M. indica (13.6?±?2.4?units?g^?1 fresh wt.). A. indica exhibited maximum guaiacol peroxidase activity (0.024?±?0.006?units?min?g^?1 fresh wt.), while S. cumini showed maximum SOD (12.5?±?2.3?units?g^?1 fresh wt.) and catalase activities (6.9?±?2.2?units?g^?1 fresh wt.). M. indica and S. cumini have been found to be more potent antioxidant systems among the studied plants.     

A rapid determination of histamine and 3-methylhistamine in human plasma by high performance liquid chromatography

Histamine plays an important pathophysiological role in allergy, inflammation, gastric acid secretion, microcirculation and neurotransmission. 3-Methylhistamine is a prominent metabolite of histamine. Different methods for determination of histamine in biological fluids have been developed. In the present study, a simple, simultaneous determination of histamine and 3-methylhistamine by HPLC (precolumn derivatization with fluorescamine) was developed in human plasma using fluorescence detection with 1-methylhistamine as the internal standard. Linear regression analysis of the ratios of the concentrations of histamine and 3-methylhistamine (X) against peak height ratios (Y) yielded the following: y = 0.0073x ? 0.0096 (R ² = 0.990) and y = 0.0077x ? 0.0111 (R ² = 0.989). In conclusion, it was possible to detect histamine and 3-methylhistamine below 5 ng mL^?1 in 1 mL plasma.     

Genotoxic assessment of anthracene and benzo [a] pyrene to milkfish Chanos chanos

Mutagenic and genotoxic effects of polycyclic aromatic hydrocarbons, anthracene and benzo [a] pyrene (BaP), in milkfish Chanos chanos were determined using micronucleus (MN) test and comet assay (CA). Distinct mean frequencies of nuclear abnormalities such as MNs; binucleated micronuclei, nuclear bud, and fragmented apoptotic cells were measured. Significant increase in DNA damage with five classes of damage level was observed and expressed in terms of arbitrary unit (AU). Mean frequencies of total nuclear abnormalities were 0.5?±?0.25 cells in control; 0.67?±?0.33 cells in solvent control; 70?±?9.60 cells in 0.176?mg?L^?1 anthracene, and 91.83?±?6.25 cells in 0.031?mg?L^?1 BaP. The greatest DNA damage of 170AU was observed in 0.176?mg?L^?1 anthracene-exposed group and 182AU was observed in 0.031?mg?L^?1 BaP-treated fish. This study confirmed that the CA and MN assays are useful tools in determining potential genotoxicity of water pollutants and might be appropriate as a part of monitoring program.     

Application of an inclusion complex for determination of dithianon residues in water and fruits

A 3,3′-benzidine/β-cyclodextrin fluorescence derivative (BCDFD) was prepared and characterized by means of elemental analysis and ¹H NMR. An inclusion complex was formed between BCDFD and the pesticide dithianon based on the internal cavity of β-cyclodextrin (β-CD), and the binding constant (K_S ) of the inclusion complex was investigated. The inclusion complex could be applied for the determination of dithianon residues. Based on this, a sensitive method for the determination of dithianon was established. The linear dynamic range was from 0.63 to 12.5?mg?L^?1 with a correlation coefficient of 0.9927. The limit of detection for an aqueous solution of standard and the relative standard deviation (RSD) were 10.6?µg?L^?1 and 0.76%, respectively. The proposed method has been successfully applied for the determination of dithianon residues in water and fruit samples with satisfactory results, and recoveries in the range of 96.0–108% were obtained. The method is rapid, direct, economical, and sensitive for dithianon analysis.     

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.