Mixtures of veterinary medicinal compounds in manured soils

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1 Workshop Pharmaceuticals in Soil, Sludge and Slurry Mixtures of veterinary medicinal compounds in manured soils Nadine Tauchnitz Daniela Gildemeister, Silvia Berkner Dessau-Roßlau, 18th June to 19th June 2013

2 Input of veterinary pharmaceuticals into soil Application of pharmaceuticals animals at pasture livestock farms Excretion urine, faeces Storage liquid (slurry) or solid manure (dung) Photochemical degradation Pasture land Transfer into plants Fertilisation Field Chemical and microbial degradation Run-off Leaching Surface water Groundwater Drinking water Mineralisation Accumulation (modified after Stroh, 2005; Stevens, 2009) -1- Background

3 Input of veterinary pharmaceuticals into soil kind and amount of pharmaceuticals animal metabolism kind and amount of fertilisers matrix properties of fertilisers storage of fertilisers application techniques application times soil tillage system/incorporation of fertilisers Transfer into plants Chemical and microbial degradation metabolites Mineralisation Accumulation Run-off Leaching Surface water Groundwater Drinking water Physicochemical properties of pharmaceuticals degradability adsorption capacity (K d ) lipophilicity (K ow, K oc ) polarity and others Soil properties mineral content and composition content of organic matter ionic strength ph-value temperature and others -2- Background

4 Veterinary pharmaceuticals in environmental samples Antibiotics tetracyclines (chlortetracycline) sulfonamides (sulfadiazine) trimethoprim fluoroquinolones (enrofloxacin) macrolides (tylosin) Antiparasitics benzimidazoles (flubendazol) avermectines (ivermectin) Organic fertiliser Soil Antibiotics tetracyclines (chlortetracycline) sulfonamides (sulfadimidin) Antiparasitics benzimidazoles (flubendazol) Surface water Groundwater Antibiotics tetracyclines (chlortetracycline) sulfonamides (sulfadimidin) trimethoprim fluoroquinolones (enrofloxacin) macrolides (tylosin) Antiparasitics benzimidazoles (flubendazol) avermectines (ivermectin) -3- Background

5 Mixtures of veterinary pharmaceuticals Parent substances + metabolites = Mixture of multicomponents interactions o synergistic o additive o antagonistic Increase or decrease of the mixtures toxicity compared to single substances Underestimation of the environmental risk Requirement of the predicted environmental concentration (PEC soil ) to assess mixtures toxicity EMA guideline on pharmaceutical fixed combination products: Consideration of separate active substances (single substances) as basis for data on fixed combination products EMA: European Medicines Agency PEC soil(mix) : addition of PEC soil of all single substances in the mixture -4- Background

6 Difficulties in the assessment of environmental risk of veterinary pharmaceuticals mixtures: no central registration of the consumption of veterinary pharmaceuticals limitation of analytical methods for the quantification of pharmaceutical residues in soils (non-extractable residues, sequestration) Aim of the present study: Estimation of the exposure of veterinary pharmaceutical mixtures by anonymous questionnaire surveys of farmers Main objectives: o quantification of the amount of applied veterinary pharmaceutical agents per farm for the slurry storage period o calculation of the expected active agent concentration of pharmaceuticals in slurry (worst case, without metabolism in animals) o exposure assessment of pharmaceutical agents by calculation of predicted environmental concentration in soil (PEC soil ) -5- Objectives

7 Methods o anonymous questionnaire surveys of farmers o duplicates of the regular forms on drug application to agricultural animals for one year ( 57 AMG, German Drug Law, documentation requirement) o calculation of the expected agents concentration in slurry without metabolism in animals PEC slurry = agents in slurry storage time/applied slurry amount o calculation of the predicted environmental concentration in soil (PEC soil ) in µg/kg according to the EMA guidelines 1 PEC soil = N max * PEC slurry N slurry * soil depth * bulk density N max : N slurry : Soil depth: 170 kg N/ha (max. N application according to fertiliser ordinance DüV) 5,3 kg/m³ (N content slurry pig fattening), 3,8 kg/m³ (N content slurry dairy farming) 0,05 bzw. 0,2 m, Bulk density: 1500 kg/m³ 1 EMEA (European Medicines Agency), CVMP (Committee for Medicinal Products for Veterinary Use). Revised Guideline on Environmental Impact Assessment for Veterinary Medicinal Products. In Support of the VICH-Guidelines GL 6 and GL 38. EMEA/CVMP/ERA/418282/2005-Rev. 1. (2008). -6- Methods

8 Example - duplicates of the regular forms on drug application to animals -7- Methods

9 Example - excel sheet for data evaluation Time period Columns A B C D E F G H I J K L M N O P Q R S identification classification agent amount amount C w1 C w2 C w3 C w1 C w2 C w3 animals C Gw1 C Gw2 C Gw3 date applicationanimals duration ml g mg/ml* mg/ml mg/ml mg mg mg amount mg mg mg category days mg/kg mg/kg mg/kg Amoxycilllin ß-Lactam Amoxycilllin i.m. F 5 Amoxycilllin ß-Lactam Amoxycilllin i.m. F 5 Amoxycilllin ß-Lactam Amoxycilllin i.m. F 5 Amoxycilllin ß-Lactam Amoxycilllin i.m. F 4 Amoxycilllin ß-Lactam Amoxycilllin i.m. F 4 Amoxycilllin ß-Lactam Amoxycilllin i.m. F 4 Amoxycilllin ß-Lactam Amoxycilllin i.m. F 6 Baycox (5 %) Kokzidienmittel Toltrazurilum oral F 2 Baycox (5 %) Kokzidienmittel Toltrazurilum oral F 2 Baycox (5 %) Kokzidienmittel Toltrazurilum oral F 2 Baycox (5 %) Kokzidienmittel Toltrazurilum oral F 2 Baycox (5 %) Kokzidienmittel Toltrazurilum oral F 2 Bisolvon Bronchosekretolytika Bromhexinhydrochlorid oral MS 5 Bisolvon Bronchosekretolytika Bromhexinhydrochlorid oral JS 5 Cobactan (2,5 %) ß-Lactam Cefquinonum i.m. F 2 Cobactan (2,5 %) ß-Lactam Cefquinonum i.m. F 2 Synutrim Sulfonamid SDZ, Trim. 0, , E oral F 7 Synutrim Sulfonamid SDZ, Trim. 0, E oral F 7 Synutrim Sulfonamid SDZ, Trim. 0, ,2 85, E oral F 7 Synutrim Sulfonamid SDZ, Trim. 0, , E oral F 7 Synutrim Sulfonamid SDZ, Trim. 0, , E oral F 7 Marbocyl 10 % Fluorquinolon Marbofloxacinum i.m. MS 2 Marbocyl 10 % Fluorquinolon Marbofloxacinum i.m. MS 2 Marbocyl 10 % Fluorquinolon Marbofloxacinum i.m. MS 2 Marbocyl 10 % Fluorquinolon Marbofloxacinum i.m. MS 2 Marbocyl 10 % Fluorquinolon Marbofloxacinum i.m. MS 2 Powerflox Fluorquinolon Enrofloxacin i.m. F 3 Powerflox Fluorquinolon Enrofloxacin i.m. F 3 Powerflox Fluorquinolon Enrofloxacin i.m. F 3 = D (E)*F = D (E)*G = I (J)*L*S -7- Methods

10 distribution of alltogether 30 questionnaires reply of 10 5 of them were fattening usable farm for the (1173 assessment animals) production targets: dairy cows, fattening pigs Application of veterinary pharmaceuticals in a pig 0.3% 1.6% 0.04% antibiotics hormones antiparasitics antiphlogistics analgetics bronchosecretogogues 97.9% -8- Results

11 Application of antibiotic agents in a pig fattening farm (1173 animals) 3.3% 4.1% 21.7% 0.3% 70.7% 0.02% beta-lactams fluoroquinolones sulfonamides/trimethoprim tetracyclines aminoglycosides polypeptides -9- Results

12 Application of veterinary pharmaceuticals in a dairy farm (750 animals) 0.01% 98.8% 0.68% 0.53% antibiotics hormones antiphlogistics analgetics bronchosecretogogues sedatives -10- Results

13 Application of antibiotic agents in a dairy farm (750 animals) 17.4% 0.6% 6% 0.3% 0.7% 74.9% beta-lactams tetracyclines fluoroquinolones aminoglycosides amphenicoles macrolides -11- Results

14 Amount of consumed pharmaceuticals (kg) Annual course of the administered veterinary pharmaceuticals (pig fattening farm) bronchosecretogogues antiparasitics hormones analgetics antiphlogistics antibiotics -12- Results

15 Annual course of the administered antibiotics (pig fattening farm) 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0% beta-lactams aminoglycosides sulfonamides/trimethoprim fluoroquinolones tetracyclines polypeptides -13- Results

16 Predicted Environmental Concentration in slurry and soil (pig fattening farm) PEC slurry PEC slurry PEC slurry PEC soil (5cm depth) PEC soil (20 cm depth) [µg L -1 ] [µg L -1 ] [µg L -1 ] [µg kg -1 ] [µg kg -1 ] April August average antibiotics beta-lactams fluoroquinolones aminoglycosides tetracyclines sulfonamides trimethoprim polypeptides hormones oxytocin progestagens gonadotropin prostaglandin agonists antiphlogistics excretion flunixin metabolism 1.03 meloxicam degradation 0.07 dexamethasone antiparasitics ivermectin toltrazurilum analgetics test organisms metamizole bronchosecretogogues bromhexine hydrochloride PEC Mix > triggervalue 100 µg/kg Environmental risk assessment has to proceed to phase II physical-chemical properties environmental behaviour effects on different aquatic and terestrial Results

17 Predicted Environmental Concentration in slurry and soil (dairy farm) PEC slurry PEC slurry PEC slurry PEC slurry PEC soil (5cm depth) PEC soil (20 cm depth) [µg L -1 ] [µg L -1 ] [µg L -1 ] [µg L -1 ] [µg kg -1 ] [µg kg -1 ] May July Oct. average antibiotics beta-lactams fluoroquinolones aminoglycosides tetracyclines amphenicoles macrolides hormones gonadotropin prostaglandin agonists antiphlogistics dexamethasone flunixin carprofen meloxicam analgetics metamizole PEC Mix no environmental risk Results

18 Maximum of 42 chemical substances Overview of administered chemical substances chemical substance substance classes therapeutic classes Amoxicillin beta-lactams antibiotics Ampicillin beta-lactams antibiotics Oxacillin beta-lactams antibiotics Penicillin beta-lactams antibiotics Procain-Penicillin beta-lactams antibiotics Benzathin-Benzylpenicillin beta-lactams antibiotics Benzylpenicillin Procain beta-lactams antibiotics Dihydrostreptomycinsulfat beta-lactams antibiotics Benzylpenicillin-Kalium beta-lactams antibiotics Neomycin beta-lactams antibiotics Cefaperazon beta-lactams antibiotics Cefapirin beta-lactams antibiotics Cefquinonum beta-lactams antibiotics Ceftiofurum beta-lactams antibiotics Ceftiofur-Natrium beta-lactams antibiotics Cloxacillin beta-lactams antibiotics Cloxacillin-Natrium beta-lactams antibiotics Cloxacillin-Benzathin beta-lactams antibiotics Oxytetracyclin tetracyclines antibiotics Tylosin macrolides antibiotics Danofloxacin fluoroquinolones antibiotics Enrofloxacin fluoroquinolones antibiotics Gentamycin aminoglycosides antibiotics Lincomycin aminoglycosides antibiotics Neomycin aminoglycosides antibiotics Streptomycin aminoglycosides antibiotics Dihydrostreptomycin aminoglycosides antibiotics Cloprostenol prostaglandin agonists hormones Trometamolum prostaglandin agonists hormones Butylscolopamin parasympathicolytic antispasmodics Buserelin peptide hormones Flunixin non-steroidal anti inflammatory drugs antiphlogistics Carprofen non-steroidal anti inflammatory drugs antiphlogistics Meloxicam non-steroidal anti inflammatory drugs antiphlogistics Dexamethason glucocorticosteroids antiphlogistics Diazepam benzodiazepine sedatives Xylazinhydrochlorid thiazine amines sedatives Embutramid hydroxybutyrate narcotics Florfenicol amphenicoles antibiotics Ketoprofen non-steroidal anti inflammatory drugs analgetics Metamizol non-opioid analgesics analgetics Methylsalicylat non-opioid analgesics analgetics Results

19 application of maximal 42 different chemical substances and 16 different substance classes in farms surveyed (primarily antibiotics: 82 to 100 %) markedly higher consumption of veterinary drugs in pig fattening compared to dairy farming pig fattening farms mainly used not easily degradable, environmentally relevant antibiotics (tetracyclines, sulfonamides/trimethoprim) with PEC soil markedly exceeding trigger value of 100 µg/kg for pig slurry calculation of PEC soil(mix) higher than1000 µg/kg and furthermore application of substance classes which was already found in environmental samples suggests: a high probability that mixtures of different veterinary drug components may occur in manured soils requirement of continued, more representative data collection (central digital registration of drug application) -17- Summary

20 Thanks for your attention!

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