The influence of gill and liver metabolism on the predicted bioconcentration of three pharmaceuticals in fish

The potential for xenobiotic compounds to bioconcentrate is typically expressed through the bioconcentration factor (BCF), which has gained increased regulatory significance over the past decade. Due to the expense of in vivo bioconcentration studies and the growing regulatory need to assess bioconcentration potential, BCF is often calculated via single-compartment models, using K(OW) as the primary input. Recent efforts to refine BCF models have focused on physiological factors, including the ability of the organism to eliminate the compound through metabolic transformation. This study looks at the ability of in vitro biotransformation assays using S9 fractions to provide an indication of metabolic potential. Given the importance of the fish gill and liver in metabolic transformation, the metabolic loss of ibuprofen, norethindrone and propranolol was measured using rainbow trout (Oncorhynchus mykiss) and channel catfish (Ictalurus punctatus) gill and liver S9 fractions. Metabolic transformation rates (k(M)) were calculated and integrated into a refined BCF model. A significant difference was noted between BCF solely based on K(OW) and BCF including k(M). These studies indicate that the inclusion of k(M) in BCF models can bring predicted bioconcentration estimates closer to in vivo values.     

Prediction of bioconcentration factors under non-equilibrium conditions

With the bioconcentration of lipophilic compounds by aquatic organisms when equilibrium between water and biota is attained, the logarithm of the bioconcentration factor has a direct linear relationship with the logarithm of the octanol/water partition coefficient. In practice, however, equilibrium may not be reached, particularly for more lipophilic compounds. It is shown theoretically, and confirmed by use of published experimental results on fish, that a different direct linear relationship also exists between the logarithms of the bioconcentration factor and the partition coefficient for compounds not achieving equilibrium after a specific exposure time. Thus, non-equilibrium bioconcentration factors can be predicted from their partition coefficients. The constants in the non-equilibrium linear equation are related to those from the relationships between equilibrium bioconcentration factors and the partition coefficients, and between the first-order clearance rate constants and the partition coefficients.     

Accumulation and elimination kinetics of di-, tri- and tetra chlorobiphenyls by goldfish after dietary and aqueous exposure

Bioaccumulation kinetics of five di-, tri- and tetrachlorobiphenyls from water and food were studied in laboratory experiments with goldfish (Carassius auratus). First order rate constants for uptake from water and clearance were determined after simultaneous administration of the five compounds in constant concentration, and were related to bioconcentration factors obtained in a static fish-water equilibration system. Biomagnification by retention of the PCB's from food was studied in a separate experiment.The difference in clearance rates for the chlorobiphenyls is the main reason for the different bioconcentration and biomagnification factors.Absorption efficiencies from water and food are higher than 40%. Clearance half lives vary from 10 days for 2,5-dichlorobiphenyl to 60 days for 2,3′,4′5-tetrachlorobiphenyl, which is correlalated with the decreasing aqueous solubilities of the compounds. Bioconcentration factors are between 0.4 × 10⁶ and 1.5 × 10⁶, biomagnification factors between 0.2 and 1.7, based on extractable lipids. Substitution of chlorine in the position para to the phenyl-phenyl bond influences hydrophobicity and bioaccumulation of the PCB's more strongly than substitution in ortho position.A kinetic model is developed which accounts for the influence of the lipid content of the fish on the clearance rate of a chemical. Reproducible determination of the bioconcentration potential of environmental chemicals is possible by use of an “internal bioaccumulation standard” in a kinetic test system. Food chain accumulation in fish is likely to be an important process only for persistent chemicals with extremely low water solubility.     

Bioaccumulation of cadmium by the freshwater isopod Asellus aquaticus (L.) from aqueous and dietary sources

Experiments were conducted to determine the kinetics and relative importance of aqueous and dietary uptake of cadmium by the freshwater isopod Asellus aquaticus (L.). Test animals were exposed during 30 days to aqueous Cd in a continuous flow system (exposure levels: 0.2 - 10 microg litre(-1)) and kept on a diet of previously contaminated Elodea sp. (range of Cd concentrations: 2-350 microg g(-1), dry weight). Preceding semi-static experiments on dosage-control of the dietary factor revealed a rapid uptake of Cd by Elodea, with relatively high concentration factors (CF), which ranged from 4.8 to 5.5 (dry weight log (CF) after 16 days). For Asellus uptake from water appeared to be the predominant route. Highly significant bioconcentration of cadmium from water was observed in the animals, even at exposure levels below 1.0 microg litre(-1). In the various treatments, direct uptake from water accounted for 50-98% of the body burdens after 30 days exposure. The experimental results were described with a first order one-compartment bioaccumulation model. Model parameter estimates (mean +/- standard error) were obtained for rate constant of uptake (560 +/- 110 day(-1)), rate constant of elimination (0.032 +/- 0.017 day(-1)) and assimilation efficiency of Cd uptake from food (1.1 +/- 0.7%). The (dry weight) bioconcentration factor (BCF) and bioaccumulation factor (BAF) for extrapolated steady state conditions were estimated at 18 000 (BCF) and 0.08 (BAF). Experiments conducted at two different pH levels (5.9 versus 7.6) revealed no significant effects of pH on the uptake of aqueous Cd by the isopods. The results are discussed in relation to their potential significance to the field situation.     

No enhancement in bioconcentration of organic contaminants by low levels of DOM.

The aim of the present work was to systematically study the effect of low concentrations of dissolved organic matter (DOM) on the bioconcentration of organic contaminants, in order to show whether the phenomenon of enhanced bioconcentration factors (BCFs), that has been reported in the literature, is generally found at low levels of DOM or if BCF enhancements are more likely due to a random scatter in the experimental data. The first part of the study tested the hypothesis that low levels of DOM affect the uptake kinetics of organic contaminants, leading to transient enhancements of BCFs, relative to DOM-free controls, which could have been reported as BCF enhancements in short-term studies. We found that the presence of low concentrations of two different types of DOM consistently decreased the bioconcentration of benzo[a]pyrene (BaP) in the water flea Daphnia magna at all exposure times (1-24 h), and that no transient BCF enhancements occurred. The second part of the study systematically investigated if low concentrations of DOM from a wide range of different aquatic systems can cause enhancements in the bioconcentration of organic contaminants. Water fleas were exposed to combinations of four different organic contaminants (BaP, tetrachlorobiphenyl, pentachlorophenol and naphthalene) with low concentrations of 12 different types of DOM that had been collected from various regions throughout Europe. In several of the DOM treatments, we found mean BCFs being higher than mean BCFs in the controls (especially for naphthalene). This shows that the experimental setup used in this study (and similarly in previous studies) can produce seeming BCF enhancements at low concentrations of DOM. However, statistical analyses showed that treatment means were not significantly different from control means. Thus, this systematic study suggests that the BCF enhancements that have been reported in the literature are more likely the result of random, experimental variations than the result of a systematic enhancement of bioconcentration.     

Unique bioconcentration characteristics of new aryl fluoroalkyl ethers in common carp (Cyprinus carpio)

The bioconcentration factors (BCFs) of seven new aryl fluoroalkyl ethers--four bis-4-tetrafluoroethoxyphenyl-type (bis-type) compounds and three mono-4-tetrafluoroethoxyphenyl-type (mono-type) compounds--were obtained by bioconcentration tests using common carp. The BCFs of 4 of the 7 ethers were higher than 5000, indicating their high bioconcentration potential. The bioconcentration characteristics of the bis-type compounds were different from those of the mono-type compounds and non-fluoro diphenylmethanes with a similar skeleton structure to the bis-type compounds, in taking longer to reach a plateau and having a slower elimination rate and in their distribution patterns in the fish body. The BCF of 1 bis-type compound was much higher than the value predicted by an accepted correlation equation between BCF and P(ow). In addition, the logP(ow) of the bis-type compounds calculated by commercially available computer software was remarkably different from that measured.     

Bioaccumulation kinetics and bioconcentration factor of chlorinated pesticides in tissues of Puntius ticto (Ham.)

Bioaccumulation kinetics and bioconcentration factor (BCF) of chlorinated pesticides like Aldrin, Dieldrin, Benzene hexachloride (BHC), and Dichlorodiphenyl-dichloro-ethane (DDT) in fish tissues of Puntius ticto was studied in detail in a continuous fed system. The bioconcentration process is summarized by using a first order uptake model and the steady-state BCF is calculated based on the 30 days exposure. Rate of bioaccumulation of DDT was maximum of 4.6432 microg g(-1) wet weight per day in liver tissue whereas it was minimum of 0.0002 microg g(-1) wet weight per day in case of Dieldrin in the muscle tissue among the pesticides. It was observed that DDT showed maximum BCF of 89.010 in case of liver tissue of the fish exposed to 30 days. The regression coefficient (r2) between pesticide concentration and exposure time varied between 0.6212 and 0.9817 indicating high correlation. Based on actual calculated BCF values, the octanol water partition coefficient (Kow) values were predicted. In order to prove the hydrophobic property of chlorinated compounds and its affinity towards lipid, the Kow is predicted. Results showed that pesticide burden differ from tissue to tissue and can be correlated to the lipid content, size, exposure time, and species.     

Bioaccumulation studies of organochlorinated pesticides in tissues of Cyprinus carpio

Freshwater fish Cyprinus carpio was selected for the study of bioaccumulation of organochlorinated pesticides in tissues like gills, muscle, intestine, kidney, and liver in a continuous fed system. The pesticides used were Aldrin, Dieldrin, BHC, and DDT. The bioaccumulation of Dieldrin was maximum of 85.0 microg g(-1) wet weight in liver tissue while minimum of 7.30 microg g(-1) wet weight for DDT at 30 days exposure time. Bioconcentration factor (BCF) has followed the same trend in liver tissue for Dieldrin and DDT. The rate of bioaccumulation was found to be maximum of 4.3879 microg g(-1) wet weight in liver tissue and minimum of 0.0021 microg g(-1) wet weight in gill tissue for 30 days exposure. As evidenced by the increasing values of BCF, pesticide uptake also showed increased trend with the increase in exposure time. A high correlation coefficient ranging between 0.7247 and 0.9616 between the pesticide concentration and exposure time was observed. Based on actual BCF values, log Kow were calculated and the values are well within the reported values of 6.5 indicating efficient relationship between BCF and log Kow because beyond the 6.5 the bioconcentration levels off.     

Fragment constant method for prediction of fish bioconcentration factors of non-polar chemicals

A fragment constant method for prediction of fish bioconcentration factor (BCF) was established based on experimental BCF values for 80 non-polar chemicals from nine classes. The model was evaluated using coefficients of determination and mean residuals, which are 0.995 and 0.1836, respectively. Jackknife tests were applied to examine the robustness of the prediction model on a class-by-class basis.     

Approach for extrapolating in vitro metabolism data to refine bioconcentration factor estimates

National and international chemical management programs are assessing thousands of chemicals for their persistence, bioaccumulative and environmental toxic properties; however, data for evaluating the bioaccumulation potential for fish are limited. Computer based models that account for the uptake and elimination processes that contribute to bioaccumulation may help to meet the need for reliable estimates. One critical elimination process of chemicals is metabolic transformation. It has been suggested that in vitro metabolic transformation tests using fish liver hepatocytes or S9 fractions can provide rapid and cost-effective measurements of fish metabolic potential, which could be used to refine bioconcentration factor (BCF) computer model estimates. Therefore, recent activity has focused on developing in vitro methods to measure metabolic transformation in cellular and subcellular fish liver fractions. A method to extrapolate in vitro test data to the whole body metabolic transformation rates is presented that could be used to refine BCF computer model estimates. This extrapolation approach is based on concepts used to determine the fate and distribution of drugs within the human body which have successfully supported the development of new pharmaceuticals for years. In addition, this approach has already been applied in physiologically-based toxicokinetic models for fish. The validity of the in vitro to in vivo extrapolation is illustrated using the rate of loss of parent chemical measured in two independent in vitro test systems: (1) subcellular enzymatic test using the trout liver S9 fraction, and (2) primary hepatocytes isolated from the common carp. The test chemicals evaluated have high quality in vivo BCF values and a range of logK(ow) from 3.5 to 6.7. The results show very good agreement between the measured BCF and estimated BCF values when the extrapolated whole body metabolism rates are included, thus suggesting that in vitro biotransformation data could effectively be used to reduce in vivo BCF testing and refine BCF model estimates. However, additional fish physiological data for parameterization and validation for a wider range of chemicals are needed.     

Measuring bioconcentration factors and rate constants of chemicals in aquatic organisms under conditions of variable water concentrations and short exposure time

A novel method, based on iterative numerical integration, is presented for deriving bioconcentration factors and rate constants of chemicals in aquatic organisms from experimental data of bioconcentration tests in which the chemical concentration in the water is variable over time and the test duration is too short to reach steady-state. The method is applied to reported data from fish and plant bioconcentration tests. The results demonstrate that this method can derive bioconcentration factors and rate constants with considerably less experimental error than other methods currently used, thus reducing uncertainty and variability in bioconcentration measurements.     

Relationship between the lipid content of fish and their bioconcentration potential of 1,2,4-trichlorobenzene

A significant positive correlation between the lipid content of eight fish species and their bioconcentration factor (BCF) of 1,2,4-trichlorobenzene is demonstrated. The log BCF value on a lipid basis is in good agreement with the log n-octanol/water partition coefficient.     

Bioconcentration of superlipophilic persistent chemicals

According to present understanding, persistent superlipophilic chemicals — such as octachlorodibenzo-p-dioxin, octachlorodibenzofuran, Mirex etc — with log K_ow > 6 and cross sections > 9.5 Å, bioconcentrate in aquatic organisms only little from ambient water. The most convincing argument against it is that in bioconcentration experiments with superlipophilic chemicals amounts applied exceeded water solubility by several orders of magnitude. This paper describes various methods for determining bioconcentration factors (BCF) of superlipophilic compounds. As exemplified with octachlorodibenzo-p-dioxin, BCF values evaluated by these methods match well with those calculated by QSARs for fish and mussels based on log K_ow and water solubility. As expected, these BCF values exceed previous values by several orders of magnitude. For BCF evaluation of superlipophilic chemicals in aquatic organisms we recommend:

1. flow-through systems, kinetic method (OECD guideline No. 305 E)
2. ambient concentrations < water solubility
3. during the uptake and especially during the elimination phase no toxic effects of the test organisms should occur.

     

Pesticide bioconcentration modelling for fruit trees

The model presented allows simulating the pesticide concentration evolution in fruit trees and estimating the pesticide bioconcentration factor in fruits. Pesticides are non-ionic organic compounds that are degraded in soils cropped with woody species, fruit trees and other perennials. The model allows estimating the pesticide uptake by plants through the water transpiration stream and also the time in which maximum pesticide concentration occur in the fruits. The equation proposed presents the relationships between bioconcentration factor (BCF) and the following variables: plant water transpiration volume (Q), pesticide transpiration stream concentration factor (TSCF), pesticide stem-water partition coefficient (K(Wood,W)), stem dry biomass (M) and pesticide dissipation rate in the soil-plant system (k(EGS)). The modeling started and was developed from a previous model "Fruit Tree Model" (FTM), reported by Trapp and collaborators in 2003, to which was added the hypothesis that the pesticide degradation in the soil follows a first order kinetic equation. The FTM model for pesticides (FTM-p) was applied to a hypothetic mango plant cropping (Mangifera indica) treated with paclobutrazol (growth regulator) added to the soil. The model fitness was evaluated through the sensitivity analysis of the pesticide BCF values in fruits with respect to the model entry data variability.     

Accumulation and excretion of chloroanilines by carp

Accumulation and excretion of chloroanilines were studied for carp ( L.). The average bioconcentration factors (BCF) in the whole body of the fish after 24 – 336 hr exposure were 2.0 and 3.7 for o-chloroaniline. 0.8 and 2.2 for m-chloroaniline and 0.8 and 1.7 for p-chloroaniline at high and low concentrations of the chemicals, respectively. The excretion rate constants (k) from the whole body of the fish were 0.19 hr⁻¹ for o-chloroaniline, 0.21 hr⁻¹ for m-chloroaniline and 0.16 hr⁻¹ for p-chloroaniline. The experimental data on the accumulation and excretion of the chloroanilines were consistent with the field data.     

Accumulation and excretion of organophosphorous pesticides by willow shiner

Bioconcentration and excretion of 8 organophosphorous pesticides were studied for willow shiner ( ). The average bioconcentration factors (BCF) in the whole body of the fish after 24 – 168 hr exposure were 0.8 for dichlorvos, 76 for salithion, 18 for methidathion, 29 for pyridaphenthion, 481 for fenthion and 36 for phosmet, Further, the BCF values of the other pesticides after 168 hr exposure were 713 for phenthoate and 1682 for EPN. The correlation between n-octanol-water partition coefficients (P_OW) and BCF in willow shiner was investigated for 19 pesticides studied here and already reported. The correlation factor (r) was not so high (0.6819, n=19) but higher (0.9085, n=18) in case excluding captan. The excretion rate constants (k) from the whole body of willow shiner were 0.20 hr⁻¹ for salthion, 0.05 hr⁻¹ for phenthoate, 0.27 hr⁻¹ for methidathion, 0.20 hr⁻¹ for pyridaphenthion, 0.07 hr⁻¹ for fenthion, 0.04 hr⁻¹ for EPN and 0.28 hr⁻¹ for phosmet.     

Bioaccumulation Kinetics and Bioconcentration Factor of Chlorinated Pesticides in Tissues of Puntius ticto (Ham.)

Abstract

Bioaccumulation kinetics and bioconcentration factor (BCF) of chlorinated pesticides like Aldrin, Dieldrin, Benzene hexachloride (BHC), and Dichloro-diphenyl-dichloro-ethane (DDT) in fish tissues of Puntius ticto was studied in detail in a continuous fed system. The bioconcentration process is summarized by using a first order uptake model and the steady-state BCF is calculated based on the 30 days exposure. Rate of bioaccumulation of DDT was maximum of 4.6432 µg g^?1 wet weight per day in liver tissue whereas it was minimum of 0.0002 µg g^?1 wet weight per day in case of Dieldrin in the muscle tissue among the pesticides. It was observed that DDT showed maximum BCF of 89,010 in case of liver tissue of the fish exposed to 30 days. The regression coefficient (r ²) between pesticide concentration and exposure time varied between 0.6212 and 0.9817 indicating high correlation. Based on actual calculated BCF values, the octanol–water partition coefficient (K _ow) values were predicted. In order to prove the hydrophobic property of chlorinated compounds and its affinity towards lipid, the K _ow is predicted. Results showed that pesticide burden differ from tissue to tissue and can be correlated to the lipid content, size, exposure time, and species.     

Bioconcentration of ibuprofen in fathead minnow (Pimephales promelas) and channel catfish (Ictalurus punctatus)

Pharmaceutical products and their metabolites are being widely detected in aquatic environments and there is a growing interest in assessing potential risks of these substances to fish and other non-target species. Ibuprofen is one of the most commonly used analgesic drugs and no peer-reviewed laboratory studies have evaluated the tissue specific bioconcentration of ibuprofen in fish. In the current study, fathead minnow (Pimephales promelas) were exposed to 250 μg L⁻¹ ibuprofen for 28 d followed by a 14 d depuration phase. In a minimized bioconcentration test design, channel catfish (Ictalurus punctatus) were exposed to 250 μg L⁻¹ for a week and allowed to depurate for 7 d. Tissues were collected during uptake and depuration phases of each test and the corresponding proportional and kinetic bioconcentration factors (BCFs) were estimated. The results indicated that the BCF levels were very low (0.08-1.4) implying the lack of bioconcentration potential for ibuprofen in the two species. The highest accumulation of ibuprofen was observed in the catfish plasma as opposed to individual tissues. The minimized test design yielded similar bioconcentration results as those of the standard test and has potential for its use in screening approaches for pharmaceuticals and other classes of chemicals.