nRtfocrlSo f 1 Ik ,1 Biomonitoring of human populations exposed to petroleum fuels with special consideration of the role of benzene as a genotoxic component Report of the EC Environment programme Project EV5V-CT92-0221 Edited by A. Carere and R. Crebelli ISSN 0394-9311 Serie Relazioni 97/4 DISCLAIMER Portions of this document may be illegible electronic image products. Images are produced from the best available original document. iSTITUTO SUPERIORS Dl SANITA Biomonitoring of human populations exposed to petroleum fuels with special consideration of the role of benzene as a genotoxic component Report of the EC Environment programme Project EV5V-CT92-0221 Edited by Angelo Carere and Riccardo Crebelli Laboratorio di Tossicologia Comparata ed Ecotossicologia ISSN 0394-9311 Serie Relazioni 97/4 Istituto Superiore di Sanita Biomonitoring of human populations exposed to petroleum fuels with special consideration of the role of benzene as a genotoxic component Report of the EC Environment programme. Project EV5V-CT92-0221 Edited by Angelo Carere and Riccardo Crebelli 1997, iv, 102 p. Sene Relazioni 97/4 In the framework of an EC research programme on the health risks of environmental chemicals, the Istituto Superiore di Sanita co-ordinated, in 1993-1996, a project on the biological effects of benzene and petroleum fuels. Seven laboratories from six European countries collaborated in the biological monitoring of selected populations with occupational exposure to petrochemicals. Several markers of early biological effect were applied together with environmental and personal exposure monitoring techniques. An epidemiological retrospective mortality study was also carried out on Italian filling station attendants. TTie results obtained highlighted an excess of genetic damage in some of the study populations, compared to matched unexposed controls. Even though these results do not allow a reliable risk estimation, the possible prognostic significance of cytogenetic damage for future cancer onset, together with some alerting findings from the mortality study, suggest that low dose exposures to benzene and petroleum fuels may retain some toxicological significance. Key words: Benzene, Biomonitoring, Epidemiology, Exposure monitoring, Petroleum fuels Istituto Superiore di Sanita Monitoraggio biologico di popolazioni esposte a carburanti, con particolare riguardo agli effetti del benzene. Rapporto del programma di ricerca comunitario Ambiente. Progetto EV5V-CT92-0221. A cura di Angelo Carere e Riccardo Crebelli 1997, iv, 102 p. Serie Relazioni 97/4 (in inglese) Nell’ambito del programma comunitario Ambiente, 1’Istituto Superiore di Sanita ha coordinate un progetto triennale (1993-1996) sugli effetti della esposizione a benzene e carburanti autoveicolari. Sette laboratory in sei stati europei, hanno partecipato al monitoraggio di popolazioni con esposizione occupazionale a bassi liveili di prodotti petrolchimici. Sono stati analizzati molteplici indicator! di effetti biologic! precoci, con particolare riguardo ai danni cromosomici, insieme a marcatori di esposizione interna, validati e intercalibrati alio scopo. I risultati ottenuti mostrano, in alcune delle popolazioni studiate, un eccesso di danni cromosomici rispetto a popolazioni di contralto non esposte. Una indagine sulle cause di morte di una coorte di gestori di impianti di rifornimento ha segnalato un possibile eccesso di neoplasie. Anche se i risultati ottenuti non permettono una puntuale stima del rischio, in considerazione del possibile valore predittivo delle alterazioni citogenetiche per i! rischio di tumore, e delle indicazioni dello studio di mortalita, si pud ritenere che anche 1’esposizione a basse dost di benzene e altri derivati del petrolio possa avere rilevanza tossicologica. Parole chiave: Benzene, Biomonitoraggio, Carburanti, Epidemiologia, Esposizione The research project “Biomonitoring of human populations exposed to petroleum fuels with special consideration on the role of benzene as a genotoxic component ” was partially supported by the EC Environment programme under the contract EV5 V-CT92-0221, coordinated by the Istituto Superiore di Sanita. The interest and encouragement of Dr Canice Nolan, Directorate General XII, Science, Research and Development of the European Commission, is gratefully acknowledged. The Editors are indebted with Ms. Francesca Di Bari for her valuable assistance. © Istituto Superiore di Sanita 1997 1 CONTENTS Preface iii Contributors iv 1. Overview of the project 1 2. Assessment of benzene exposure, early genetic effects, and cancer 6 mortality in Italian filling station attendants 3. Cytogenetic monitoringof Spanish filling station attendants 41 4. Benzene exposure and cytogenetic investigation on Estonian shale oil 56 petrochemical workers 5. Influence of benzene and benzene ralated compounds on cytogenetic 68 damage in human blood lymphocytes (Polish workers) 6. Examination of ras oncoproteins in human plasma from healthy controls 78 and workers exposed to petroleum emissions 7. Monitoring of Hungarian urban areas, oil refinery sites and service 86 stations 8. Conclusions 96 9. Publications arising from the project 99 Ill PREFACE Benzene is nowadays an ubiquitary environmental pollutant in European countries. Engine emissions supply the largest contribution to the environmental benzene level, which is a matter of special concern in many high traffic urban areas. Despite the long standing interest of physicians and biologists in the adverse effects of benzene to human health, the consequences of human exposure to low environmental benzene levels are not yet elucidated. This leads to uncertainties in the estimation of actual risks and in the definition of air quality criteria. In 1991 the European Commission launched the “Environment” Programme, to support coordinated research activities of European laboratories in the field of the environment. Airborne pollution, and the resulting health risks to urban populations, were among the priority tasks. In this framework, the Istituto Superiore di Sanita coordinated a collaborative research project on exposure and early biological effects of low benzene exposure. Six partners from five countries, including two non- Member States, joined the project from March 1993 to September 1996. The main results and conclusions of this collaborative endeavour are summarized in this publication. Exposure and early effect markers were assessed on several model populations with moderate to low benzene exposure, providing information on low dose effects which contribute to unravelling the health risks from environmental benzene. The project also provided the opportunity for a fruitful exchange of expertise and collaboration among partners, giving a truly transnational dimension to this coordinated effort, which tackles a priority environmental health problem throughout the European community. The Editors IV CONTRIBUTORS Angelo Carere, Riccardo Crebelli, Andrea Zijno, Paola Leopardi, Francesca Marcon, Cristina Andreoli, Luigi Turrio Baldassarri, - Laboratory of Comparative Toxicology and Ecotoxicology, Istituto Superiore di Sanita, Rome, Italy Susanna Lagorio, Ivano Iavarone, Sergio Fuselli - Laboratory of Environmental Hygiene, Istituto Superiore di Sanita, Rome, Italy Francesco Forastiere, Elisabetta Rapid - Epidemiological Unit, Latium regional Health Authority, Rome, Italy Marco Biocca, Adriana Pasquini - CDS, Centro Documentazione per la Salute, Emilia Romagna Health Authority, Bologna, Italy Antonio Antoccia, Daniela Cimini, Francesca Degrassi, Mario Fiore, Fabrizio Palitti, Antonella Sgura, Caterina Tanzarella - Centro di Studio di Genetica Evoluzionistica del CNR c/o Dipartimento di Genetica e Biologia Molecolare, Universita “La Sapienza ”, Rome, Italy Ricardo Marcos, Amadeo Creus, Noel Xamena, Elisabet Carbonell, Maria Pitarque, Gloria Ribas - Universitat Autonoma de Barcelona, Barcelona, Spain Kimmo Peltonen, Antero Aitio, Tiina Anttinen-Klemetti, Kirsi Autio, Tehri Kuljukka, Lars Nylund, Kaija Pekari, Jordi Surralles, Maqa Sorsa - Finnish Institute of Occupational Health, Helsinki, Finland Diana Anderson, Jane Hughes - BIBRA International, Carshalton, Surrey, United Kingdom Antonina Cebulska-Wasilewska, Anna Wierzevvska, Ewa Kasper - Institute of Nuclear Physics, Kracow, Poland Alan Pinter, Anna Paldy, Eva Vaskovi, Julianna Bacskai, Gabor Mayer, Istvan Vincze - National Institute of Public Health, Budapest, Hungary Thomas Veidebaum - Estonian Institute of Experimental and Clinical Medicine, Tallin, Estonia 1 1. OVERVIEW OF THE PROJECT Introduction Benzene, an established human carcinogen (IARC, 1987 Vol. 1), is currently the fourth-fifth (by tonnes) organic chemical produced worldwide (Fishbein L., 1988). Benzene is an important air pollutant in most large Western and Eastern cities, where it is detectable at levels in the range of tens of micrograms per cubic meter (Fishbein L., 1988 - Wallace L.A., 1989). Epidemiological data on leukemia incidence in benzene exposed workers have been used to derive the unit risk, i.e. the risk for a lifetime exposure to 1 microgram per cubic meter. Current estimates are in the range 4 x 10"6 - 3 x 10"5 the unit risk for leukemia associated to benzene exposure (OMS/WHO, 1987 - EPA, 1994). These figures have been used recently by the Italian National Advisory Committee on Toxicology to make a quantitative estimate of leukemia risks associated with benzene exposure in the general Italian population living in urban areas: according to the hypothesized exposure levels,