Impact of microcystin containing diets on physiological performance of Nile tilapia (Oreochromis niloticus) concerning detoxification

Nile tilapia (Oreochromis niloticus) were fed by diets supplemented with cyanobacteria containing in part the cyanotoxin microcystin-LR (MC-LR) to determine the potential impacts on detoxification. Four different diets were prepared based on a commercial diet: (1) control, (2) MC-5% (containing 5% dried Microcystis sp. biomass with 4.92 μg MC-LR g(-1) diet), (3) MC-20% (containing 20% dried Microcystis sp. biomass with 19.54 μg MC-LR g(-1) diet), and (4) Arthr-20% (containing 20% dried Arthrospira sp. biomass without MC-LR). Blood and liver samples were taken after one, 7, and 28 days and protein has been determined in plasma and liver. In the liver, impacts on detoxification were measured by glutathione-S-transferase (GST) activities and gene expression of multi drug resistance protein (MDRP). Plasma protein did not change between all four diets at any sampling time whereas liver protein was significantly elevated already after one day in Arthr-20% and after 28 days in both, MC-20% and Arthr-20%. Biochemical measurements of GST activities revealed no significant impact at any sampling time. In order to characterize the potential effect of MC-LR on MDRP, RT-qPCR method was established. However, as for GST activities no significant changes in MDRP gene expression have been observed. Thus, in summary, oral exposure of MC-LR containing cyanobacteria to Nile tilapia via feed ingestion did not impact significantly detoxification in liver concerning GST activities and MDRP expression despite biochemical composition concerning liver protein was significantly elevated by the diets containing 20% cyanobacteria biomass, regardless whether they contained MC-LR or not.     

Use of biomarkers in Nile tilapia (Oreochromis niloticus) to assess the impacts of pollution in Bolgoda Lake, an urban water body in Sri Lanka

The present study reports the first analysis of water pollutants in Sri Lankan waters using a suite of biomarkers in Nile tilapia (Oreochromis niloticus) residing in Bolgoda Lake which receives urban, industrial and domestic wastes from multiple sources. The fish were collected from the lake in the dry period (April 2005) and wet periods (September 2005, October 2006) and the levels of biomarkers viz. hepatic ethoxyresorufin O-deethylase (EROD), glutathione S-transferase (GST), metallothioneins, biliary fluorescent aromatic compounds, brain and muscle cholinesterases (ChE) were compared with those of the laboratory reared control fish and the fish obtained from a less polluted water body, Bathalagoda reservoir (reference site). The results revealed that biomarker levels of the fish collected from the reference site were not significantly different from the controls. Hepatic EROD and GST activities in fish from Bolgoda Lake were induced 4.2-16.6 folds and 1.4-3.3 folds respectively compared with the control fish. Analysis of bile in the lake fish revealed recent uptake of naphthalene, pyrene and benzo(a)pyrene type polycyclic aromatic hydrocarbons (PAHs). The induction of EROD activities in feral fish reflects the exposure of fish to aryl hydrocarbon receptor agonists including PAHs present as pollutants in the Bolgoda Lake. Cholinesterase activity in the fish inhabiting one sampling site of Bolgoda Lake was lower (22-40% inhibition) than the activity measured in the control fish indicating the presence of anticholinesterase pollutants in the area. Hepatic metallothionein levels in the lake fish were higher (1.9-3.2 folds) in comparison to the controls indicating metal exposure. The results support the potential use of these biomarkers in Nile tilapia in assessing pollution in tropical water bodies.     

Oxidative stress response in zebrafish (Danio rerio) gill experimentally exposed to subchronic microcystin-LR

The worldwide occurrence of cyanobacterial blooms makes it necessary to perform environmental risk assessment procedures to monitor the effects of microcytins on fish. Oxidative stress biomarkers are valuable tools in this regard. Considering that zebrafish (Danio rerio) is a common model species in fish toxicology and the zebrafish gill is potentially useful in screening waterborne pollutants, this study investigated the oxidative stress response in zebrafish gill exposed to subchronic microcystin-LR (MCLR) concentrations (2 or 20?μg/l) via measurement of toxin accumulation, protein phosphatase (PP) activity, and the antioxidant parameters (glutathione-S-transferase-GST; glutathione-GSH; superoxide dismutase-SOD; catalase-CAT; glutathione peroxide-GPx; glutathione reductase-GR), as well as levels of hydroxyl radical (OH) and lipid peroxidation (LPO). The results showed that after 30?days exposure, MCLR accumulated in zebrafish gill and MCLR exposure induced PP activity in gill. A linear inhibition of GST activity and GSH content was observed in the gills, revealing that they were involved in the first step of MCLR detoxification. The 2?μg/l MCLR treatment neglectably affected OH content and the antioxidant enzymes (SOD, CAT, GPx, and GR), however oxidative stress was induced under the 20?μg/l MCLR treatment in which an enhanced OH content and alterations of the antioxidant enzymes were observed in the treated gills, although both treatments exerted little effect on LPO level. The principal component analysis results indicated that the most sensitive biomarkers of MCLR exposure were GST and GSH in zebrafish gill. So, D. rerio could be regarded as a suitable bioindicator of MCLR exposure by measuring CAT, GR, GST, and GSH as biomarkers.     

Detection of polybrominated diphenyl ethers in tilapia (Oreochromis mossambicus) from O'ahu, Hawai;i

Polybrominated diphenyl ethers (PBDEs) have been detected for the first time at a range from 231.58 to 685.61 ng g(-1) lipid weight in the muscles of tilapia (Oreochromis mossambicus) collected from O'ahu, an island of the geographically isolated Hawaiian archipelago.     

Assessment of metal status in drainage canal water and their bioaccumulation in Oreochromis niloticus fish in relation to human health

The purpose of this study was to assess metal concentrations (Al, Cd, Pb, Hg and Ni) in Sabal drainage canal (Al-Menoufiya Province, River Nile Delta, Egypt) water as well as their accumulation in some selected organs (skin, muscles and kidneys) of Oreochromis niloticus fish to evaluate their hazard levels in relation to the maximum residual limits for human consumption. Drainage canal water was found to be heavily polluted with metals which far exceeded the permissible limits. It was found that metals accumulated in organs of O. niloticus in concentrations higher than those of canal water. Kidneys of O. niloticus contained the highest concentrations of the detected metals, while skin appeared to be the least preferred site for the bioaccumulation of metals as the lowest metals concentrations were detected in this tissue. The present study shows that fish organs contained high levels of metals exceeding the permissible limits values. Metals in muscle of fish were higher than the maximum permissible concentrations for human consumption. Thus, consuming fish caught from drainage canals is harmful to the consumers.     

Coelomocyte biomarkers in the earthworm Eisenia fetida exposed to 2,4,6-trinitrotoluene (TNT)

Contamination by 2,4,6-trinitrotoluene (TNT) is a global environmental problem at sites of former explosive production, handling, or storage, and could have deleterious consequences for human and ecological health. We investigated its sublethal effects to Eisenia fetida, using two nonspecific biomarkers. In coelomocytes of earthworms exposed 24, 48, or 72 h, we evaluated DNA damage (comet assay) and neutral red retention time (NRRT), using the filter paper contact test. Both percentage of damage (D%) and calculated damage index showed significant DNA damage at almost all concentrations, at all time points assayed. Along exposure time, two different patterns were observed. At the lower TNT concentrations (0.25?C0.5 ??g/cm²) an increased DNA migration at 48 h, with a decrease close to initial levels after 72 h exposure, was observed. This decrease could be attributed to activation of the DNA repair system. At higher concentrations (1.0?C2.0 ??g/cm²), the high DNA damage observed remained constant during the 72 h exposure, suggesting that the rate of DNA repair was not enough to compensate such damage. Analysis of NRRT results showed a significant interaction between time and treatment. After 48 h, a significant decrease was observed at 4.0 ??g/cm². After 72 h, NRRT presented a concentration-dependent decrease, significantly different with respect to control at 0.5, 1.0, 2.0, and 4.0 ??g/cm². The two assayed methods, performed on the same sample, showed clear responses to sublethal TNT exposure in E. fetida, providing sensitive unspecific biomarkers of cell injury and DNA damage.     

Oxidative stress,glutathione level and antioxidant response to heavy metals in multi-resistant pathogen,Candida tropicalis

In this study, we explored the multiple heavy metal-resistant yeast isolated from heavy metal-polluted environment. The isolated yeast showed maximum growth at 30 °C, pH 7.0, and the strain was identified as Candida tropicalis through 18S ribosomal RNA (rRNA) gene sequence analysis. Yeast cells grew well in medium containing different concentrations of heavy metal ions [CdCl₂, Pb(NO₃)₂, NaAsO₂, CuSO₄ and K₂Cr₂O₇]. Minimum inhibitory concentration (MIC) against different metal ions was ranged from 5 to 19 mM, and the metal resistance value against each metal observed by yeast cells was 5 mM (Cr), 10 mM (Cd), 15 mM (As), 14 mM (Cu) and 19 mM (Pb) and increased in the following order: Pb > Cu > As ≥ Cd > Cr. The total cellular glutathione, GSH/GSSG redox couple and metallothioneins like protein (MT) were assayed by growing cultures for 24 h and exposed to 100 mg/L of each heavy metal ion. Remarkable increase in γ-glutamylcysteinylglycine (GSH) level was determined in arsenic and cadmium treatment followed by chromium, lead and copper. Stressed cells had much more oxidized GSH than unstressed cells. GSH/GSSG ratio was significantly increased in cadmium and copper treatment in contrast to chromium, arsenic and lead. Statistical analysis revealed significantly higher cysteine level in all metal-treated samples as compared to control. Antioxidant glutathione transferase activity was not detected in metal-treated and untreated yeast samples. One-dimensional electrophoresis of proteins revealed marked differences in banding pattern of heavy metal-exposed yeast samples. A prominent 20 kDa band was observed in all treated samples suggesting that some differential proteins could be over-expressed during heavy metal treatment and might be involved in cell resistance mechanisms.     

Heat shock protein 47 stress responses in Chinese hamster ovary cells exposed to raw and reclaimed wastewater

As wastewater reclamation and reuse becomes more widespread, risks of exposure to treated wastewater increase. Moreover, an unlimited number of pollutants can be identified in wastewater. Therefore, comprehensive toxicity assessment of treated wastewater is imperative. The objective of this study was to perform a comprehensive toxicity assessment of wastewater treatment systems using stress response bioassays. This powerful tool can comprehensively assess the toxicity of contaminants. In this study, samples from conventional activated sludge treatment, membrane bioreactors (MBRs) with different pore sizes and sludge retention times (SRTs), rapid sand filtration, coagulation, nano-filtration (NF) and reverse osmosis (RO) were investigated. The results of stress response bioassays confirmed that the secondary effluent showed higher stress response than influent indicating that biological treatment generates toxic compounds. The results obtained from molecular weight fractionation of water samples demonstrated that organic matter with a higher molecular weight fraction (>0.1 μm) causes toxicity in secondary effluent. Furthermore, supernatant from MBR reactors showed toxicity regardless of SRT. On the other hand, stress response was not detected in MBR permeates except for an MBR equipped with a larger pore size membrane (0.4 μm) and with a short SRT (12 days). While rapid sand filtration could not remove the toxic compounds found in secondary effluent, coagulation tests, operated at an appropriate pH, were effective for reducing stress response in the secondary effluent. Experimental findings also showed that stress response was not detected in cases of NF and RO permeate subsequent to MBR treatment.     

Modulations in the levels of respiration and ions in carp Cyprinus carpio (L.) exposed to cypermethrin

This paper reports an investigation of the sublethal toxicity of cypermethrin (20 µg/l) on the levels of respiration and selected ions in tissues of carp (Cyprinus carpio L.). A significant decrease in O ₂ consumption and ions was observed in gill, brain, liver and muscle tissues of cypermethrinexposed fish. The decreased rate of O ₂ consumption may be due to inhibition of oxidative enzymes during cypermethrin stress.     

Alteration of antioxidant enzymes and impairment of DNA in the SiO2 nanoparticles exposed zebra fish (Danio rerio)

The incorporation of nanoparticles in industrial and biomedical applications has increased significantly in recent years, yet their hazardous and toxic effects have not been studied extensively. While standard toxicological test methods are generally capable of detecting the toxic effects, the choice of relevant methods for nanomaterials is still discussed. Among the various oxide nanomaterials, silica nanoparticles are widely used in biological applications that include nano-medicine. But studies on adverse effects of silica nanoparticle exposure to fish remain unclear. Therefore, the present study was designed to investigate the oxidative toxic effects of silicon dioxide nanoparticles using fish model. The size of the SiO₂ nanoparticles was between 68 and 100 nm which was confirmed by X-ray diffractometer, dynamic light scattering, scanning electron microscope and transmission electron microscope. The zebra fish were exposed to sub-lethal concentrations (5 and 2.5 mg/L) of characterized SiO₂ nanoparticles for a period of 7 days. After 7 days, SiO₂ nanoparticle-treated fishes were sacrificed, and tissues such as liver, muscle and gill were dissected out for the analysis of antioxidant enzymes and DNA fragmentation. The DNA profiles were analysed in the tissues of zebra fish that treated with SiO₂ nanoparticles. Tissues of fish from clean water were used as control, and DNA profiles were analysed. It is found that DNA from control tissues was intact, whereas the tissues treated with SiO₂ were all fragmented. SiO₂ nanoparticle-mediated antioxidant enzymes activities, such as catalase, superoxide dismutase, glutathione (GSH)-S-transferase, glutathione reductase and GSH, in the tissues of zebra fish were measured. The results revealed that alteration of antioxidant enzymes due to SiO₂ nanoparticle can be considered as a biomarker to SiO₂-mediated oxidative stress in biological samples.     

Genotoxic effects and gene expression in <Emphasis Type="Italic">Danio rerio</Emphasis> (Hamilton 1822) (Cypriniformes: Cyprinidae) exposed to mining-impacted tributaries in Manizales,Colombia

The zebrafish (Danio rerio) is one of the most studied aquatic organisms for water biomonitoring, due to its sensitivity to environmental degradation and resistance to toxic substances. This study determined the presence of micronuclei and nuclear abnormalities in peripheral blood erythrocytes, and assessed the gene expression of caspase-3 (CASP-3) and metallothionein 1 (MT-1) in the gills and liver of D. rerio. The study fish (n = 45) were exposed to water collected from two stations with mining impact (E2 and E3) and a reference station without evident mining contamination (E1), all located in La Elvira stream (Manizales-Colombia). In addition, a positive control (PC) with HgCl₂ (50 μg/L) and negative control (NC) with tap water were included. The fish from the PC and E2 and E3 treatments displayed genotoxic effects and changes in gene expression, with significant differences in micronuclei formation and the presence of blebbed nuclei. The cytochrome oxidase subunit I (COI) gene was used as reference and proved to be stable compared to the β-actin and 28S ribosomal RNA (28S) genes. In gills, CASP-3 expression was higher in the PC, and MT-1 expression was higher in the PC and E3 treatment. In liver, CASP-3 was expressed in the E2 treatment, and MT-1 expression was low. These results show that the genotoxic effects and differential gene expression observed in fish exposed to water from La Elvira stream could also be affecting the organisms present in this habitat.     

Evaluation of mentum deformities of Chironomus spp. (Chironomidae: Diptera) larvae using modified toxic score index (MTSI) to assess the environmental stress in Juru River Basin, Penang, Malaysia

Morphological mentum deformities which represent sublethal effect of exposure to different types of pollutants were evaluated in Chironomus spp. larvae inhabiting three polluted rivers of Juru River Basin in northwestern peninsular Malaysia. Using mentum deformity incidences, the modified toxic score index (MTSI) was developed based on Lenat??s toxic score index (TSI). The suggested MTSI was compared with TSI in terms of its effectiveness to identify different pollutants including heavy metals. The MTSI showed stronger relationship to total deformity incidence expressed as percentage. Additionally, the multivariate RDA model showed higher capability of MTSI to explain the variations in heavy metal contents of the river sediments. The MTSI was recommended in bioassessment of water and sediment quality using the mentum deformities of Chironomus spp. larvae from aquatic ecosystems receiving anthropogenic, agricultural, or industrial discharges.     

Method for automatic determination of soybean actual evapotranspiration under open top chambers (OTC) subjected to effects of water stress and air ozone concentration

The present study describes an operational method, based on the Katerji et al. (Eur J Agron 33:218-230, 2010) model, for determining the daily evapotranspiration (ET) for soybean inside open top chambers (OTCs). It includes two functions, calculated day par day, making it possible to separately take into account the effects of concentrations of air ozone and plant water stress. This last function was calibrated in function of the daily values of actual water reserve in the soil. The input variables of the method are (a) the diurnal values of global radiation and temperature, usually measured routinely in a standard weather station; (b) the daily values of the AOT40 index accumulated (accumulated ozone over a threshold of 40 ppb during daylight hours, when global radiation exceeds 50 Wm(-2)) determined inside the OTC; and (c) the actual water reserve in the soil, at the beginning of the trial. The ensemble of these input variables can be automatable; thus, the proposed method could be applied in routine. The ability of the method to take into account contrasting conditions of ozone air concentration and water stress was evaluated over three successive years, for 513 days, in ten crop growth cycles, excluding the days employed to calibrate the method. Tests were carried out in several chambers for each year and take into account the intra- and inter-year variability of ET measured inside the OTCs. On the daily scale, the slope of the linear regression between the ET measured by the soil water balance and that calculated by the proposed method, under different water conditions, are 0.98 and 1.05 for the filtered and unfiltered (or enriched) OTCs with root mean square error (RMSE) equal to 0.77 and 1.07 mm, respectively. On the seasonal scale, the mean difference between measured and calculated ET is equal to +5% and +11% for the filtered and unfiltered OTCs, respectively. The ability of the proposed method to estimate the daily and seasonal ET inside the OTCs is therefore satisfactory following inter- and intra-annual tests. Finally, suggestions about the applications of the proposed method for other species, different from soybean, were also discussed.     

Assessment of thermal stress adaptation by monitoring Hsp70 and MnSOD in the freshwater gastropod,Bellamya bengalensis (Lamark 1882)

Expression of the stress biomarkers 70-kDa heat shock proteins (Hsp70) and manganese superoxide dismutase (MnSOD) was measured as the molecular basis of adaptive response against increased experimental temperatures (32–40 °C for a span of 24–72 h) on the fresh water molluscan species, Bellamya bengalensis (Lamark 1882). The experimental snail specimens were collected during summer and winter seasons from two contrasting wetlands: an ecorestored (free from human interference) site (SI) and other experiencing anthropogenic stresses (SII). The mortality rate of the B. bengalensis and the immunoblotting of MnSOD and Hsp70 of their digestive glands were performed at regular intervals during the period of heat stress. The SI provided a lower stress environment based on physicochemical parameters such as pH, dissolved oxygen (DO), biological oxygen demand (BOD), chemical oxygen demand (COD), and alkalinity for the survival of test species, although both sites experienced mortality due to thermal stresses. The parity in protein expressions displayed a uniform mode of adaptive impact to temperature elevations in both field and laboratory exposure. The Hsp70 expression was minimal at lower thermal stress, but increased with a rise in temperature. It is very likely that higher Hsp70 levels are not directly related to survival or adaptation. In contrast, MnSOD levels appeared to be an indicator of adaptive responses vis-a-vis survival of the animals. So, the expression levels of a universal free radical scavenger like MnSOD are recognized as a potential biomarker in a bioindicator species like Bellamya.     

Morphometric and metabolic indicators of metal stress in wild yellow perch (Perca flavescens) from Sudbury,Ontario: a review

Eighteen lakes studied near Sudbury and across Northeastern Ontario (Canada) over a five-year period provided a wide contamination gradient of cadmium (Cd), copper (Cu) and other metals such as nickel (Ni) and zinc (Zn). All were inhabited by yellow perch (Perca flavescens), which was sometimes the only species present. Liver Cd and Cu concentrations were monitored in these lakes, in some cases for several consecutive years and for multiple seasons. This data suggests that yellow perch from clean to mildly-contaminated environments loosely control their hepatic Cu concentrations between 7 and 50 microg g dry weight(-1), and a threshold of 50 microg g dry weight(-1) is suggested as the normal range of homeostatic control. Similar data collected by others support this value. Liver Cd concentrations appeared more variable among lake samples, but consistently remained below 10 microg g dry weight(-1) in clean to mildly-contaminated lakes, also supported by data collected elsewhere. Condition factors allowed the discrimination between clean and polluted yellow perch, a conclusion consistent with data for the same species collected in the Rouyn-Noranda area (Quebec, Canada). Values of weight-to-length scaling coefficient lower than 3.0 also discriminated between clean and metal-polluted yellow perch. Finally, three studies indicated that chronic metal exposure can lead to an impairment of aerobic capacities in wild yellow perch, as indicated by lower muscle activity of citrate synthase (CS), aerobic swim performance and respiration rate. We propose that the combination of liver metal concentrations, scaling coefficient, condition factor and an indicator of physiological impairment such as muscle CS activity can provide a suitable range of parameters to adequately assess the effects of metal contamination on the health of yellow perch. Although yellow perch are ubiquitous in North America, this approach can potentially be applied to other small fish species more suitable to other study areas.     

Response mechanisms of antioxidants in bryophyte (Hypnum plumaeforme) under the stress of single or combined Pb and/or Ni

The short-term responses and mechanisms of antioxidants in moss Hypnum plumaeforme subjected to single or combined Pb and/or Ni stress has been revealed in this study, in order to clarify (1) the relationship between the stress intensity and antioxidant fluctuation, (2) the difference between single and combined stress, and (3) the possibility of biomonitoring by the application of antioxidant fluctuation under stress. The results showed that the stress induced dose dependent formation of reactive oxygen species (ROS) and subsequent lipid peroxidation. Total chlorophyll (Chl) content and superoxide dismutase (SOD) activity were initiated under lower stress but were inhibited under higher stress. Both single and combined stress decreased catalase (CAT) activity but increased peroxidase (POD) activity, indicating POD in the moss played an important role in resisting the oxidative stress induced by Pb and Ni. The accumulation of (.)O2(-) and H2O2 in H. plumaeforme was respectively related to the low activity of SOD and the decreased activity of CAT. The study indicated that Pb and Ni had synergistic effect in inducing the oxidative stress in moss H. plumaeforme, especially under the combination of high concentration of Ni (0.1, 1.0 mM) and Pb. POD and CAT activity, as well as H2O2 and MDA content, which increased or decreased regularly with a dose dependent under Pb and Ni stress, could be used as an effective indicator in moss biomonitoring, especially in the case of light pollution caused by heavy metals without the changes in the appearance of mosses.     

Spatial and temporal trends of polycyclic aromatic hydrocarbons in wild mussels from the Cantabrian coast (N Spain) after the Prestige oil spill

Polycyclic aromatic hydrocarbon (PAH) levels were determined in tissues of wild mussels (Mytilus galloprovincialis) collected at 17 stations along the Cantabrian coast (N Spain), from Navia (Asturias) to Fuenterrabía (Basque Country), in order to assess the extent of the environmental impact caused by the Prestige oil spill (November 13, 2002). Six sampling campaigns were carried out in April, June and November in 2003 and 2004. The comparison of PAH data with those obtained earlier in 2000 showed a widespread pyrolytic and petrogenic contamination and allowed an estimation, for the first time, of the background pollution in the region and identification of the chronic hotspots. The spatial distribution found in the first samples after the oil spill revealed the eastern area as the most affected due to the continuous arrival of fuel slicks since early summer 2003. Several stations in this area showed increased total PAH concentrations of up to 15 times the pre-spill levels, which did not recover until April 2004, more than one year after the accident. Molecular parameters within the aliphatic and aromatic fractions were determined to assess the presence of Prestige oil in these samples.     

Speciative determination of Cr (III) and Cr (VI) in dyeing waste water of Dil Creek discharge to Izmit Gulf (Izmit-Kocaeli, Turkey) by ICP-AES

Wastewater pollution in industrial areas is one of the most important environmental problems. Heavy metal pollution, especially chromium pollution in wastewater sources from dyeing and tannery has affected the life on earth. This pollution can affect all ecosystems and human health directly or by food chain. Therefore, the determination of chromium in this study is of great importance. Dil Creek is located in the eastern Marmara region and discharges into the Izmit Gulf. This water source is used for irrigation in agriculture and as drinking water for animals. In this study, a rapid, sensitive and selective method for the speciative direct determination of Cr (III) and Cr (VI) in dyeing waste water samples collected from the nearest station to Izmit Gulf of Dil Creek in May 2006 by inductively coupled plasma-atomic emission spectrometry (ICP-AES) has been developed. An analysis of a given sample is completed in about 15 min for ICP-AES the method. As the result of the chromium analysis, the limit of quantification (LOQ) for the Cr (III), Cr (VI) and total Cr were founded as 0.0111 ± 0.0002 mg/l (RSD, 1.80%), 0.0592 ± 0.0010 mg/l (RSD, 1.70%) and 0.0703 ± 0.0020 mg/l (RSD, 2.84%) respectively. In addition, the general mathematical formula has been developed to calculate the concentration of Cr(III), which can be applied to any other metal species. The result of Cr (VI) analysis indicated that water quality of Creek was IV. class quality according to the inland water classification. In order to validate the applied method, recovery studies were performed.