Journal of Hazardous Materials 176 (2010) 300–305 Contents lists available at ScienceDirect Journal of Hazardous Materials journal homepage: www.elsevier.com/locate/jhazmat Toxicity of purified terephthalic acid manufacturing wastewater on reproductive system of male mice (Mus musculus) Xu-Xiang Zhang a,∗ , Shi-Lei Sun b , Yan Zhang a , Bing Wu a , Zong-Yao Zhang a , Bo Liu a , Liu-Yan Yang a , Shu-Pei Cheng a a State Key Laboratory of Pollution Control and Resource Reuse, School of the Environment, Institute of Environmental Health & Ecological Security, Nanjing University, Nanjing, Jiangsu 210093, China b Water Research & Development Center, Jiangsu Suyuan Environemental Protection Engineering Co. Ltd, Nanjing 210093, China article info Article history: Received 22 April 2009 Received in revised form 3 November 2009 Accepted 4 November 2009 Available online 13 November 2009 Keywords: Purified terephthalic acid Reproductive toxicity Spermatogenic cells Wastewater abstract Reproductive toxicity of purified terephthalic acid (PTA) manufacturing wastewater on the male mice (Mus musculus) was investigated after 35-day intragastric perfusion treatment with the wastewater. Fluorescein diacetate and propidium iodide staining, and flow cytometry were used to assess the toxicity of PTA wastewater on spermatogenic cells. PTA wastewater induced significant variations in the relative percentages of immature haploid, diploid, tetraploid and S-phase spermatogonia. Percentage of viable spermatogenic cells was reduced from 93.1 ± 2.3 in control group to 90.4 ± 1.9 in the wastewater-treated group. Testicular histopathology revealed expansion of interstitial space and reduction in the number and size of Leydig cells induced by the wastewater, which was further certified by the decrease (10.6%) in relative testes weight and the increase (101.3%) in sperm shape abnormality in the wastewater-treated group. In this study, PTA wastewater was found to have reproductive toxicity on male mice, and public health problems may potentially arise from the discharge of the wastewater into the environment. © 2009 Elsevier B.V. All rights reserved. 1. Introduction Purified terephthalic acid (PTA) is a widely used raw material of polyester, and China is one of the main PTA producers in the world [1]. Generally, for the preparation of 1 ton of PTA, 3–10 ton of wastewater is produced during the manufacturing process [2,3]. Main compounds in PTA wastewater are terephthalic acid (TA), p- toluic acid (p-Tol), p-carboxybenzaldehyde (p-CBA), phthalic acid (PA) and benzoic acid (BA) [4,5]. Among the components in PTA wastewater, TA can cause bladder stones and bladder cancer [6,7], as well as impairment of testicular functions [8]. Information is unavailable about the reproductive toxicity induced by p-CBA, but the compound p-Tol was found to cause the decrease of epididymal weight and the increase of incidence in cauda epididymal oligo/azoospermia [9]. It is well-known that PA can disrupt endocrine function and induce reproductive and developmental toxicity in laboratory animals [10]. Generally, BA is not considered clinically as a reproductive or developmental toxicant [11], but it was still revealed that the compound could affect the growth and reproduction of freshwater organisms [12] and sperm viability and the function of accessory ∗ Corresponding author. Tel.: +86 25 83595995; fax: +86 25 83595995. E-mail address: zhangxx@nju.edu.cn (X.-X. Zhang). gonad of exposed worker [13]. Although these aromatic compounds in PTA wastewater exhibit high reproductive toxicity, so far no report has been found to address the joint effects of these pollutants in the wastewater on reproductive system of mammals. This study aims to investigate the potential reproductive toxicity of PTA wastewater on male mice in terms of alterations of testicular cell population, sperm morphology and testicular histopathology, and to provide more scientific information for responsible author- ities to make up regulatory standards and guidelines to control the discharge of PTA wastewater. 2. Materials and methods 2.1. PTA wastewater PTA wastewater samples were collected from a local wastew- ater treatment plant of SinopecYangtze Petrochemical Company Ltd. (Nanjing, China). Measurements of chemical oxygen demand (COD), biological oxygen demand in 5 days (BOD 5 ), total nitro- gen, total phosphorus and total suspended solid were carried out according to NEPAC (The National Environmental Protection Agency of China) standard methods [14]. The aromatic compounds in PTA wastewater were measured by HP 6890 gas chromatog- raphy (GC) coupled with an HP 5973 mass spectrometer (MS) (Hewlett-Packard Co., USA). A 1.0 ml of sample was introduced into 0304-3894/$ – see front matter © 2009 Elsevier B.V. All rights reserved. doi:10.1016/j.jhazmat.2009.11.028