Impact of Veterinary Pharmaceuticals on the Agricultural Environment: A Re-inspection.

The use of veterinary pharmaceuticals (VPs) is a result of growing animal production. Manure, a great crop fertilizer, contains a significant amount of VPs. The investigation of VPs in manure is prevalent, because of the potential risk for environmental organisms, as well as human health. A re-evaluation of the impact of veterinary pharmaceuticals on the agricultural environment is needed, even though several publications appear every year. The aim of this review was to collate the data from fields investigated for the presence of VPs as an inevitable component of manure. Data on VP concentrations in manure, soils, groundwater and plants were collected from the literature. All of this was connected with biotic and abiotic degradation, leaching and plant uptake. The data showed that the sorption of VPs into soil particles is a process which decreases the negative impact of VPs on the microbial community, the pollution of groundwater, and plant uptake. What was evident was that most of the data came from experiments conducted under conditions different from those in the environment, resulting in an overestimation of data (especially in the case of leaching). The general conclusion is that the application of manure on crop fields leads to a negligible risk for plants, bacteria, and finally humans, but in future every group of compounds needs to be investigated separately, because of the high divergence of properties.

Bioaccumulation and analytics of pharmaceutical residues in the environment: A review.

The presence of pharmaceutical residues in various environmental compartments is an issue of increasing concern. The widespread occurrence of these compounds in water and soil samples has been demonstrated in a number of analytical studies. However, the data about their concentrations in biota samples is scarce. Moreover, the trophic transfer of pharmaceuticals remains largely unexplored, despite increasing evidence of the potential bioaccumulation of those compounds. Therefore, the main aim of this review is to present an overview of the current state of data about the bioaccumulation and analytical methodologies used for the determination of pharmaceutical residues in biota samples. This work focuses on the most commonly found pharmaceuticals in the environment: antibiotics, analgesic and anti-inflammatory drugs, steroid hormones, antihypertensives and antidepressants. We do hope that the collected data will allow a better understanding of pharmaceutical pollution and the exposure of non-target organisms. However, although impressive progress has undoubtedly been made, in order to fully understand the behavior of these chemicals in the environment, there are still numerous gaps to be filled in our overall knowledge in this field.

The leaching behavior of cyclophosphamide and ifosfamide from soil in the presence of co-contaminant--Mixture sorption approach.

Anticancer drugs (ACDs) exhibit high biological activity, they are cytotoxic, genotoxic, and are constantly released into the environment as a result of incomplete metabolism. Consequently they pose a serious threat to the environment and human health due to their carcinogenic, mutagenic and/or reproductive toxicity properties. Knowledge of their bioavailability, including their sorption to soils and their impact on the soil-groundwater pathway, is crucial for their risk assessment. Laboratory batch and column leaching tests are important tools for determining the release potential of contaminants from soil or waste material. Batch and column tests were carried out with soils differing in physicochemical properties, each spiked with cyclophosphamide (CK) or ifosfamide (IF). Moreover, due to the fact that environmental pollutants may occur as coexisting compounds in the soil the mobility evaluation for ACDs in the mixture with metoprolol (MET; ß-blocker) as a co-contaminant was performed. In order to assess appropriateness, the batch and column tests were compared. The release depended on the properties of both the soil and the presence of co-contaminants. The faster release was observed for coarse-grained soil with the smallest organic matter content (MS soil: 90% decrease in concentration until liquid-to-solid ratio (L/S) of 0.3 L kg(-1) for all tests' layout) than for loamy sand (LS soil: 90% decrease in concentration until ratio L/S of 0.75 L kg(-1)). ACDs are highly mobile in soil systems. Furthermore, the decrease of mobility of ifosfamide was observed with the presence of a co-contaminant (metoprolol) in both of the soils (in MS soil a decrease of 29%; in LS soil a decrease of 26%). The mobility of cyclophosphamide does not depend on the presence of a contaminant for MS soil, but also exhibits a decrease of 21% in LS soil.

Sorption of sulfisoxazole onto soil--an insight into different influencing factors.

Although sulfonamides (SAs) are among the most commonly used veterinary drugs and their presence in the environment is well documented, knowledge of their fate and behavior in the soil environment is still limited, especially for sulfisoxazole (SSX) which is characterized by the lowest (among other SAs) pK a value associated with acid-base equilibrium of sulfonamide group. Thus, this work was focused on determining the sorption potential of SSX onto natural soils differing in physicochemical properties. All the results were modeled using linear, Freundlich, Langmuir, Dubinin-Radushkevich, and Temkin sorption isotherms. The established sorption coefficients (K(d)) for SSX were quite low (from 0.27 to 0.95 L kg(-1)), which indicated that this substance is highly mobile and has the potential to run off into surface waters and/or infiltrate ground water. The sorption data of SSX is well fitted to the Freundlich isotherm model (R(2) > 0.968). Moreover, we assessed the sorption mechanism of these compounds in the edaphic environment with respect to organic matter (OM) content, pH, and ionic strength. To clarify the current state of knowledge, these factors were examined much more thoroughly than in previous investigations concerning other SAs. The wide range of ionic strength examined showed positive correlation of this factor and sorption of SAs. The results also yielded new insight into dependency of sorption of SAs on organic matter content in soil.

Preliminary study on suitability of ionic liquids as potential passive-sampling media of polyaromatic-hydrocarbon (PAH) analyses in water.

Recently, ionic liquids (ILs) have been regarded as an attractive water-immiscible phase in liquid-liquid extraction. Because ILs have a wide range of polarity irrespective of their miscibility with water, the possibility of using them as an effective extraction phase for a broad range of contaminants means they are starting to be of particular interest. In this study we investigated a wide variety of ionic liquids, which are known to be hydrolytically stable and of a hydrophobic character, for their potential suitability as passive-sampling media for monitoring selected polyaromatic hydrocarbons. Preliminary research in this field has indicated very promising results using these novel extraction media. Because there is an enormous number of possible cation-anion combinations offering tuneable properties of ionic liquids with the potential for effective passive extraction, we hope this paper will encourage the scientific community to undertake further studies verifying the undoubted usefulness of these alternative solvents as passive samplers for many other groups of analytes. Additionally, because of the unusual solubility properties that have already been proved for ILs, it is very probable that it would soon be possible to deliver a very effective system able to extract analytes differing widely in polarity.

Toxicity of anthelmintic drugs (fenbendazole and flubendazole) to aquatic organisms.

Flubendazole (FLU) and fenbendazole (FEN) belong to benzimidazoles-pharmaceuticals widely used in veterinary and human medicine for the treatment of intestinal parasites as well as for the treatment of systemic worm infections. In recent years, usage of these drugs increased, which resulted in a larger contamination of the environment and possible negative effects on biota. Hence, in our research, we investigated an aquatic ecotoxicity of these pharmaceuticals towards: marine bacteria (Vibrio fischeri), green algae (Scenedesmus vacuolatus), duckweed (Lemna minor) and crustacean (Daphnia magna). Ecotoxicity tests were combined with chemical analysis in order to investigate the actual exposure concentration of the compounds used in the experiment as well as to stability and adsorption studies. As a result, study evaluating sensitivity of different aquatic organisms to these compounds and new ecotoxicological data is presented. The strongest negative impact of FLU and FEN was observed to D. magna.

Critical points in the evaluation of analytical methods based on liquid chromatography separation for the determination of doramectin in different environmental samples.

In recent years, the number of analytical methods of target compound residues (such as pharmaceuticals) has grown rapidly. Most of them are based on high performance liquid chromatography (HPLC). From the economic point of view, it is usual to apply the conditions of available HPLC methods or to design extraction and chromatographic separation conditions using HPLC and transfer them subsequently to a more sensitive technique like liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). However, if such a transfer is planned, it is important to assess the quality of the newly-designed LC-MS/MS method. The determination of parameters like matrix effects (ME), extraction efficiency (EE) and absolute recovery (AR) is mandatory. These parameters can visualise the weakest step in the analytical method and enable methods based on different techniques to be compared. The aim of this work was to show how quality assessment should be carried out in order to transfer an optimised method from one technique to another. The representative compound used in our investigation was doramectin (DOR), an anthelmintic drug used in veterinary medicine. The quality of the suggested methods for determining this drug in three environmental matrices (water, sediment and fish tissue) using HPLC-UV and LC-MS/MS was evaluated on the basis of known values of absolute recovery (HPLC-UV) and matrix effect, extraction efficiency and absolute recovery (all LC-MS/MS). Finally, the suggested methods for determining DOR in water, sediment and fish tissue based on LC-MS/MS measurements were validated and applied to the analysis of real environmental samples.

A comprehensive approach to the determination of two benzimidazoles in environmental samples.

Among the various pharmaceuticals regarded as emerging pollutants, benzimidazoles--represented by flubendazole and fenbendazole--are of particular concern because of their large-scale use in veterinary medicine and their health effects on aquatic organisms. For this reason, it is essential to have reliable analytical methods which can be used to simultaneously monitor their appearance in environmental matrices such as water, sediment and tissue samples. To date, however, such methods relating to these three matrices have not been available. In this paper we present a comprehensive approach to the determination of both drugs in the mentioned above matrices using liquid chromatography-ion trap mass spectrometry (LC-MS/MS). Special attention was paid to the sample preparation step. The optimal extraction methods were further validated by experiments with spiked water, sediment and fish tissue samples. Matrix effects were established. The following absolute recoveries of flubendazole and fenbendazole were achieved: 96.2% and 95.4% from waters, 103.4% and 98.3% from sediments, and 98.3% and 97.6% from fish tissue samples, respectively. Validation of the LC-MS/MS methods enable flubendazole and fenbendazole to be determined with method detection limits: 1.6 ng L(-1) and 1.7 ng L(-1) in water samples; 0.3 ng g(-1) for both compounds in sediment samples, and 3.3 ng g(-1) and 3.5 ng g(-1) in tissue samples, respectively. The proposed methods were successfully used for analysing selected pharmaceuticals in real samples collected in northern Poland. There is first data on the concentration in the environment of the target compounds in Poland.

Determination of metronidazole residues in water, sediment and fish tissue samples.

Metronidazole (MNZ) is an antibacterial and antiprotozoal drug used in veterinary and human medicine. Its continual entry into the environment and its biological properties may have significant, long-term effects on the stability of ecosystems because MNZ and its metabolites possess mutagenic, carcinogenic and toxic properties. For this reason, the application of MNZ in food-producing species is prohibited in the EU, the USA and other countries. To ensure human food safety and to protect the environment, robust and reliable screening and confirmatory tests capable of the low-level detection of MNZ residues are required. The development of methods for MNZ determination in biological and environmental samples is thus an important analytical task in environmental and food science. This work focuses on the evaluation of a method for determining MNZ in water, sediment and fish tissue samples using liquid chromatography--ion trap mass spectrometry (LC-MS/MS). MNZ was extracted from waters on Strata XC cartridges using solid phase extraction (SPE), and from sediments and fish tissues by solid-liquid extraction (sediment: 15 mL 0.1 M HCl (pH=0.6), 15 min; fish tissue: 15 mL 1% CH3COOH in ACN, 1 min; drying: 5 g MgSO4(anhyd.; 30 s) with SPE purification of the extracts (from sediment: Strata XC cartridge; from fish tissue: Supelco NH2 cartridge). The optimal procedure that we developed was validated in order to confirm its reliability and sensitivity. Matrix effects (ME) were established. Absolute recoveries ranged from 89.3% to 97.2%, and the method detection limits were 3.4 ng L(-1) (water samples), 0.4 ng g(-1) (sediment samples) and 0.3 ng g(-1) (tissue samples). These methods were used to determine MNZ in surface waters, sediments and fish tissues from the Polish River Goscicina; MNZ was found in all these matrices. The highest concentrations in water, sediment and tissue were 136.2 ng L(-1), 12.0 ng g(-1) and 1.5 ng g(-1) respectively. The results confirmed that these methods are suitable for the simultaneous analysis of waters, sediments and fish tissues for the presence of MNZ.

Beta-blockers in the environment: part I. Mobility and hydrolysis study.

Beta-blockers (BB) are one of the most widely used pharmaceuticals whose presence in different environmental compartments has already been proven in concentrations of even up to a few µg L(-1). However, our knowledge of their fate in the environment is still scarce. To obtain a better understanding on the environmental behavior of three selected BB comprehensive laboratory experiments assessing their mobility and hydrolytic stability has been conducted. Propranolol, metoprolol and nadolol--the most commonly consumed and detected in environmental samples--were selected as representatives of this group of pharmaceuticals. The objectives of our research were: (i) evaluation of the sorption potential and an explanation of the sorption mechanisms of these compounds onto soil and clay mineral (kaolinite); and (ii) investigation of the hydrolytic stability of these BB according to OECD 111. This comprehensive study supports the Environmental Risk Assessment of these pharmaceuticals.

Development of sensitive and reliable LC-MS/MS methods for the determination of three fluoroquinolones in water and fish tissue samples and preliminary environmental risk assessment of their presence in two rivers in northern Poland.

Antibiotic consumption (e.g. fluoroquinolones (FQs)) and, as a consequence, their presence in the environment, have received a lot of attention in the last several years due to increasing numbers of diseases and infections that are becoming resistant to traditional treatments for both humans and animals. In addition, even though antibiotics are safe for human and veterinary usage, ecosystems may be exposed to these substances. In this study, analytical methods for determining enrofloxacin (ENR), norfloxacin (NOR) and ciprofloxacin (CIP) in water samples and fish tissue based on the LC-MS/MS technique were developed and validated. As there is no data available concerning the risks posed by antibiotics in Poland, the proposed methods were applied for monitoring drug presence in environmental samples collected from two rivers in northern Poland. Evaluations of the ecotoxicity of ENR, NOR and CIP towards four different species of aquatic organisms: marine bacteria (Vibrio fischeri), green algae (Scenedesmus vacuolatus), duckweed (Lemna minor) and crustacean (Daphnia magna), were also carried out. All the investigated compounds were detected at least once in the survey. NOR was found to be the most ubiquitous drug with concentrations of up to 442.8 ng L(-1). Moreover, it was established that L. minor is the most sensitive species to the investigated drugs (EC50NOR = 0.13 mg L(-1), EC50ENR = 0.22 mg L(-1) and EC50CIP = 0.34 mg L(-1)). The calculated risk quotient (RQ) values confirmed that the concentrations of the investigated FQs in the environmental samples were at a level of moderate environmental risk (1

Beta-blockers in the environment: part II. Ecotoxicity study.

The increasing consumption of beta-blockers (BB) has caused their presence in the environment to become more noticeable. Even though BB are safe for human and veterinary usage, ecosystems may be exposed to these substances. In this study, three selected BB: propranolol, metoprolol and nadolol were subjected to ecotoxicity study. Ecotoxicity evaluation was based on a flexible ecotoxicological test battery including organisms, representing different trophic levels and complexity: marine bacteria (Vibrio fischeri), soil/sediment bacteria (Arthrobacter globiformis), green algae (Scenedesmus vacuolatus) and duckweed (Lemna minor). All the ecotoxicological studies were supported by instrumental analysis to measure deviation between nominal and real test concentrations. Based on toxicological data from the green algae test (S. vacuolatus) propranolol and metoprolol can be considered to be harmful to aquatic organisms. However, sorption explicitly inhibits the hazardous effects of BB, therefore the risks posed by these compounds for the environment are of minor importance.

Thermodynamic studies for adsorption of ionizable pharmaceuticals onto soil.

Although pharmaceutical compounds (PCs) are being used more and more widely, and studies have been carried out to assess their presence in the environment, knowledge of their fate and behavior, especially under different environmental conditions, is still limited. The principle objective of the present work, therefore, is to evaluate the adsorption behavior of three ionizable, polar compounds occurring in different forms: cationic (propranolol - PRO), anionic (sulfisoxazole - SSX) and neutral (sulfaguanidine - SGD) onto soil under various temperature conditions. The adsorption thermodynamics of these researched compounds were extensively investigated using parameters such as enthalpy change (ΔH°), Gibbs free energy change (ΔG°) as well as entropy change (ΔS°). These calculations reveal that sorption of PRO is exothermic, spontaneous and enthalpy driven, sorption of SGD is endothermic, spontaneous and entropy driven whereas sorption of SSX is endothermic, spontaneous only above the temperature of 303.15K and entropy driven. Furthermore, we submit that the calculated values yield valuable information regarding the sorption mechanism of PRO, SGD and SSX onto soils.

Column and batch tests of sulfonamide leaching from different types of soil.

Sulfonamides (SAs) and their metabolites present severe hazards to human health and the environment, mainly because of antibiotic resistance. Knowledge of their bioavailability, including their sorption to soils and their impact on the soil-groundwater pathway, is crucial to their risk assessment. Laboratory batch and column leaching tests are important tools for determining the release potential of contaminants from soil or waste materials. Batch and column tests were carried out with soils differing in particle size distribution, organic matter content and pH, each spiked with sulfonamides (sulfadimethoxine (SDM), sulfaguanidine (SGD), sulfisoxazole (SX)). In order to test the applicability of leaching tests to polar contaminants batch and column tests were also compared. In the column tests, release was found to depend on the properties of both soil and sulfonamides. The fastest release was observed for coarse-grained soil with the smallest organic matter content (MS soil; 100% decrease in concentration until liquid-to-solid ratio (L/S) of 0.9 L kg(-1) for all SAs). The slowest release was established for sulfadimethoxine (24.5% decrease in concentration until L/S 1.22 L kg(-1)). The results of the batch and column tests were comparable to a large extent, with slightly higher concentrations being obtained in the column test experiments of fine-grained soils with a high organic matter content.

Aquatic toxicity of four veterinary drugs commonly applied in fish farming and animal husbandry.

Doramectin (DOR), metronidazole (MET), florfenicol (FLO), and oxytetracycline (OXT) are among the most widely used veterinary drugs in animal husbandry or in aquaculture. Contamination of the environment by these pharmaceuticals has given cause for concern in recent years. Even though their toxicity has been thoroughly analyzed, knowledge of their ecotoxicity is still limited. We investigated their aquatic toxicity using tests with marine bacteria (Vibrio fischeri), green algae (Scenedesmus vacuolatus), duckweed (Lemna minor) and crustaceans (Daphnia magna). All the ecotoxicological tests were supported by chemical analyses to confirm the exposure concentrations of the pharmaceuticals used in the toxicity experiments, since deviations from the nominal concentration can result in underestimation of biological effects. It was found that OXT and FLO have a stronger adverse effect on duckweed (EC50=3.26 and 2.96mgL(-1) respectively) and green algae (EC50=40.4 and 18.0mgL(-1)) than on bacteria (EC50=108 and 29.4mgL(-1)) and crustaceans (EC50=114 and 337mgL(-1)), whereas MET did not exhibit any adverse effect in the tested concentration range. For DOR a very low EC50 of 6.37×10(-5)mgL(-1) towards D. magna was determined, which is five orders of magnitude lower than values known for the toxic reference compound K2Cr2O7. Our data show the strong influence of certain veterinary drugs on aquatic organisms and contribute to a sound assessment of the environmental hazards posed by commonly used pharmaceuticals.

Hydrolysis of sulphonamides in aqueous solutions.

Hydrolysis is one of the most common reactions controlling abiotic degradation and is one of the main paths by which substances are degraded in the environment. Nevertheless, the available information on this process for many compounds, including sulphonamides (a group of antibiotic drugs widely used in veterinary medicine), is very limited. This is the first study investigating the hydrolytic stabilities of 12 sulphonamides, which were determined according to OECD guideline 111 (1st category reliability data on the basis of regulatory demands on data quality for the environmental risk assessment of pharmaceuticals). Hydrolysis behaviour was examined at pH values normally found in the environment. This was prefaced by a discussion of the acid-base properties of sulphonamides. All the sulphonamides tested were hydrolytically stable at pH 9.0, nine (apart from sulphadiazine, sulphachloropyridazine and sulphamethoxypyridazine) were stable in this respect at pH 7.0 and two (sulphadiazine and sulphaguanidine) at pH 4.0 (hydrolysis rate≤10%; t(0.5 (25°C))>1 year). The degradation products were identified, indicating two independent mechanisms of this process. Our results show that under typical environmental conditions (pH and temperature) sulphonamides are hydrolytically stable with a long half-life; they thus contribute to the on-going assessment of their environmental fate.

Sulfadimethoxine and sulfaguanidine: their sorption potential on natural soils.

Sulfonamides (SAs) are one of the oldest groups of veterinary chemotherapeutic agents. As these compounds are not completely metabolized in animals, a high proportion of the native form is excreted in feces and urine. They are therefore released either directly to the environment in aquacultures and by grazing animals, or indirectly during the application of manure or slurry. Once released into the environment, SAs become distributed among various environmental compartments and may be transported to surface or ground waters. The physicochemical properties of SAs, dosage and nature of the matrix are the factors mainly responsible for their distribution in the natural environment. Although these rather polar compounds have been in use for over half a century, knowledge of their fate and behavior in soil ecosystems is still limited. Therefore, in this work we have determined the sorption potential of sulfadimethoxine and sulfaguanidine on various natural soils. The influence on sorption of external factors, such as ionic strength and pH, were also determined. The sorption coefficients (K(d)) obtained for the sulfonamides investigated were quite low (from 0.20 to 381.17 mL g(-1) for sulfadimethoxine and from 0.39 to 35.09 mL g(-1) for sulfaguanidine), which indicated that these substances are highly mobile and have the potential to run off into surface waters and/or infiltrate ground water. Moreover, the sorption of these pharmaceuticals was found to be influenced by OC, soil solution pH and ionic strength, with higher K(d) values for soils of higher OC and lower K(d) values with increasing pH and ionic strength.