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Although grouping and read across are currently being prac-iced, transparent guidelines to standardize the approach andacilitate consistency are needed to bolster the credibility of readcross assessments as well as provide a starting point for furtherefinement.

Recently, we published a systematic approach for identifyingnd evaluating analogs for read across assessments based uponhemical and biochemical principles (Wu et al., 2010) as well asseries of blinded case studies to test the consistency and objectiv-

ty of our framework (Blackburn et al., 2010) Based on the results ofhese case studies and our experience with use of SAR approachese contend that SAR can be successfully applied to develop pro-

ective, surrogate values for risk assessment.Successful application of our approach requires significant

xpertise as well as discipline to not overstep the boundaries ofhe defined analogs and the rating system. The end result of thisigor is that read across cannot always be supported for all end-oints and/or all chemicals. Future work will be targeted towardsxpanding read across capabilities.

oi:10.1016/j.toxlet.2012.03.073

11ollution from drug manufacturing––Assessing and managingisks in different regions of the world

11-1harmaceutical pollution from Indian drug manufacturing

oakim Larsson

University of Gothenburg, Sweden

The great majority of research on the fate and effects of pharma-euticals in the environment has so far been devoted to pollutionhrough usage. The use and excretion of drugs rarely cause morehan ng/L levels of environmental contamination. Still, for someharmaceuticals, this is sufficient for concern. However, dur-

ng the past few years, we and others have demonstrated thatirect releases from manufacturing sites around the world can

n some cases result in thousands or even a million times highernvironmental concentrations than those caused by excretion ofedicines. These findings have changed our perception for how

rugs can enter the environment, what levels may be encounterednd therefore what the associated risks may be for humans andildlife, and eventually also how pharmaceutical pollution may

e managed. Hence, during the past five years my group has putuch of our focus on identifying risks associated with pollution

rom drug manufacturing in India, particularly releases of fluoro-uinolone antibiotics. Results from several fate and effect studiesill be presented. The emphasis will be on effects on antibiotic

esistance promotion, a subject of particular relevance for publicealth, globally. This research also includes the search for novelesistance factors that we may face in the clinic tomorrow. Finally,

ome potential management strategies will be presented and dis-ussed.

oi:10.1016/j.toxlet.2012.03.075

s 211S (2012) S4–S23

S11-2Antibiotic pollution from Chinese drugmanufacturing-antibiotic resistance

Yu Zhang 1, Min Yang 1, Miaomiao Liu 1, Li Dong 1, Ren Renli 2

1 Chinese Academy of Sciences, China, 2 North China PharmaceuticalGroup Corpora, China

To investigate the effects on resistance development underlong-term antibiotic selection pressures, wastewater and biomasssamples from antibiotic production wastewater (APW) treatmentplants as well as antibiotic production wastes were taken and ana-lyzed.

Clone libraries, real-time PCR, and a high-throughput functionalgene array (GeoChip 3.0) analyses were used to evaluate the func-tional structure of microbial communities from two plants, whichtreat APW containing high concentrations of oxytetracycline (OTC).

Eleven tetracycline resistance genes (tet) and three mobile ele-ments were detected by PCR. Eight frequently reported tet genes(tet(A), tet(C), tet(L), tet(M), tet(O), tet(Q), tet(W) and tet(X)), Class1 integrons (IntI1), and transposon Tn916/1545 were successfullyquantified by real-time PCR. The relative abundances of tet genesand IntI1 in the treated effluent and activated sludge were higherthan those in the OTC production wastes, and were 1–4 log higherthan those from sewage treatment plants, showing that it may benecessary to regulate the discharge of the effluents and sludge fromAPW treatment plants. A high release of IntI1 was also found in theeffluent and sludge. According to GeoChip analysis, antibiotic resis-tance and transporter genes (559 genes) were abundant in bacteriaand gene signal intensity positively correlated with antibiotics inAPW biological systems. A total of 29.0% of the microbial commu-nity variation could be explained by antibiotics. This study providesinsights into microbial functional gene structure in APW biologicaltreatment systems, revealed the relative high antibiotic resistancegene discharge from APW biological systems.

doi:10.1016/j.toxlet.2012.03.076

S11-3Pharmaceutical formulation facilities as sources ofpharmaceuticals

Patrick Phillips 1, Steven Smith 1, Dana Kolpin 1, Steven Zaugg 1,Herbert Buxton 1, Edward Furlong 1, Kathleen Esposito 2, BeverleyStinson 2

1 US Geological Survey, United States, 2 AECOM, United States

Facilities involved in the manufacture of pharmaceutical prod-ucts are an under-investigated source of pharmaceuticals to theenvironment. Between 2004 and 2009, 35–38 effluent sampleswere collected from each of three WWTPs (wastewater treat-ment plants) in New York and analyzed for seven pharmaceuticalsincluding opioids and muscle relaxants. Two WWTPs (NY2 andNY3) receive substantial flows (>20% of plant flow) from phar-maceutical formulation facilities (PFF) and one (NY1) receives noPFF flow. Samples of effluents from 23 WWTP across the UnitedStates were analyzed once for these pharmaceuticals as part of anational survey. Maximum pharmaceutical effluent concentrationsfor the national survey and NY1 effluent samples were generally

<1 �g/L. Four pharmaceuticals (methadone, oxycodone, butalbitaland metaxalone) in samples of NY3 effluent had median concen-trations ranging from 3.4 to >400 �g/L. Maximum concentrationsof oxycodone (1700 �g/L) and metaxalone (3800 �g/L) in sam-

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les from NY3 effluent exceeded 1000 �g/L. Four pharmaceuticalsbutalbital, carisoprodol, and oxycodone) in samples of NY2 efflu-nt had median concentrations ranging from 2 to 11 �g/L. Thesendings suggest that current manufacturing practices at these PFFsan result in pharmaceuticals concentrations from 10 to 1000 timesigher than those typically found in WWTP effluents.

oi:10.1016/j.toxlet.2012.03.077

11-4dverse effects of pharmaceutical manufacture discharges inild fish

ilfried Sanchez

INERIS, France

Purpose: Adverse effects of APIs are documented in laboratoryxperiments but no data was available on the effects induced byharmaceutical factory discharge on wildlife. The present study of

ndividual and populational effects in gudgeons living in a streamubjected to a pharmaceutical factory discharge began as a con-equence of angler’s observations reporting morphological fishbnormalities such as abdominal swelling associated to gonadypertrophy in wild gudgeons living downstream from an indus-rial effluent.

Methods: To determine the origin of this disturbance, gudgeonsere electrofished in three sites located upstream and downstream

rom the industrial effluent. In all sampled fish, a set of biomark-rs including endocrine disruption end-point (vitellogenin, gonadistology), biotransformation enzymes, together with neurotoxic-

ty and immunotoxicity indicators was measured. To complete thessessment of individual effects, sex-ratio of gudgeon populationas analyzed and fish assemblage was characterized to identifyotential population disturbance.

Results and conclusions: As revealed by biotransformationnzyme responses, fish from the investigated area were exposedo organic pollutants. Downstream from the industrial effluent,sh exhibited strong signs of endocrine disruption includingitellogenin induction, intersex and male-biased sex-ratio. Thesendividual effects were associated to a decrease of density and aack of sensitive fish species. No cause-effect relationship can bermly established between fish exposure to APIs, individual andopulation disturbances, but a set of evidence supports the hypoth-sis that these compounds induce observed adverse effects andndicates that resident fish populations from both downstreamites could disappear. These results argue for the deployment ofn effect-based specific monitoring of pharmaceutical factories.

oi:10.1016/j.toxlet.2012.03.078

11-5water quality standard approach to managing

harmaceutical emissions

ichard Murray-Smith

AstraZeneca, United Kingdom

As with all chemicals used in industrial processes, emissionsf active pharmaceutical ingredients (APIs) from pharmaceutical

anufacturing facilities need to be managed effectively so that

hey do not adversely affect the local receiving environment. Thisaises the question of what emission levels of APIs would be safe.straZeneca has addressed this question by adopting the concept

s 211S (2012) S4–S23 S17

of Environmental Reference Concentrations (ERCs) and MaximumTolerable Concentrations (MTCs), to identify the long-term andshort-term concentration of an API which should not be exceeded inthe aquatic environment receiving pharmaceutical manufacturingeffluents.

The ERC/MTC approach is based on established environmen-tal quality standard concepts currently used in much nationaland international legislation, together with published method-ologies to protect the aquatic environment (algae, invertebratesand fish), top predators (fish-eating mammals such as otters)and humans. Although chronic toxicity data are preferred for ERCderivation, acute data, with appropriate considerations of uncer-tainty, may be used when chronic data are not available. Thisapproach takes all available information into account, particularlyfor older established medicines that may predate current regula-tory requirements for environmental data, and consequently helpsprioritize resources for environmental testing.

The ERC approach has been applied to 30 of AstraZenecas APIs.Merits of the approach will be discussed, together with opportuni-ties for potential future refinement.

doi:10.1016/j.toxlet.2012.03.079

S12New mechanistically based models for evaluation of druginduced liver injury: The IMI predictive DILI project

S12-1Tacking the challenge posed by drug induced liver injury: TheIMI MIP-DILI consortium

Gerry Kenna

AstraZeneca, United Kingdom

Drug induced liver injury (DILI) is a frequent cause of delayedor terminated drug development, restricted drug usage and pre-cautionary regulatory action. A key challenge is to develop novelapproaches which can be used to enable selection of drug can-didates that have reduced propensity to cause DILI in humans. Atiered test cascade is required, which addresses key chemical andcellular processes and includes in silico tools, in vitro assays, in vivomodels and translational biomarkers. Although many promisingassays, models and candidate DILI biomarkers have been described,currently there is no consensus on which approaches should beused or how they should be integrated together.

This topic has been prioritised by the EU/EFPIA InnovativeMedicines Initiative for pre-competitive research funding and aconsortium has been formed (mechanism based integrated systemsfor the prediction of drug induced liver injury: acronym MIP-DILI).This comprises 11 pharmas, 9 academic research groups and 6 SMEpartners and will tackle the problem from 2012 to 2017.

The primary goal is to identify and validate a panel of in vitro“best practice assays” for prediction of DILI in humans. Importantsupportive goals are to enhance understanding of the relationshipbetween in vitro assay signals and DILI in vivo, in animals andhumans; and to develop novel modelling approaches that inte-grate multiple preclinical data types to improve DILI prediction.An additional benefit will be enhanced understanding amongstacademia, pharmas and regulators of the value and limitations of

new and existing approaches for DILI hazard identification andrisk assessment.

doi:10.1016/j.toxlet.2012.03.081