• Abstract and Figures
• Public Full-text

Chemosphere 144 (2016) 1342e1350 Contents lists available at ScienceDirect Chemosphere journal homepage: www.elsevier.com/locate/chemosphere Vitellogenin expression in wild cyprinid Petroleuciscus esfahani as a biomarker of endocrine disruption along the Zayandeh Roud River, Iran * Neda Gilannejad a, b, , Salar Dorafshan a, Fatemeh Paykan Heyrati a, Nasrollah Mahboobi Soofiani a, Saeid Asadollah a, Juan Antonio Martos-Sitcha b, c, Francisco Prat b, Manuel Yúfera b, Gonzalo Martínez-Rodríguez b a Department of Natural Resources, Isfahan University of Technology, 84156-83111 Isfahan, Iran b Instituto de Ciencias Marinas de Andalucía (ICMAN), CSIC, 11519 Puerto Real, Cadiz, Spain c Department of Biology, Faculty of Marine Sciences, University of Cadiz, 11519 Puerto Real, Cadiz, Spain highlights An endemic cyprinid was used as a bioindicator of estrogenic exposure in the Zayandeh Roud River. vtg mRNA was detected in males from sites located downstream of anthropogenic pollution sources. The site downstream of a steel mill plant had the max vtg expression and growth impairment. article info abstract Article history: Aquatic environments are the ultimate sink for most of anthropogenic pollutants. The Zayandeh Roud Received 9 June 2015 River is the most important river in the central Iranian Plateau, supplying water to a large population. In Received in revised form order to determine the potential occurrence and in vivo effects of Endocrine Disrupting Chemicals (EDCs) 28 September 2015 with estrogenic or anti-androgenic properties we analyzed the wild populations of an extensively Accepted 29 September 2015 distributed endemic fish species, Petroleuciscus esfahani. For this purpose, specimens were caught from Available online xxx two sites upstream and two sites downstream of the expected major anthropogenic pollution sources. P. esfahani full-length cDNAs for vitellogenin (vtg), with 4177 base pairs (bp) encoding a 1339 amino acids Keywords: b fi Bioindicator (aa), and for -actin (actb), with 1776 bp encoding a 375 aa, were ampli ed and cloned. Hepatic vtg EDCs mRNA expression levels were measured by quantitative real-time PCR. Condition factor, gonadosomatic Quantitative PCR index and sex ratio were calculated and compared with vtg expression. Gonad histology was performed River ecosystem to study the possible presence of intersex condition. Detection of vtg transcripts in male individuals from the two downstream sampling sites supports the hypothesis of exposure to EDCs in these regions. Higher vtg expression in male individuals, together with reduced gonad size and condition factor, in specimens from the site located downstream of the major steel mill plant suggest a major endocrine disruption in this area. © 2015 Elsevier Ltd. All rights reserved. 1. Introduction 2011). Among them, there is a family of contaminants known generically as Endocrine Disrupting Chemicals (EDCs). EDCs with There is a growing concern about a high variety of environ- estrogenic and anti-androgenic properties affect at different levels mental pollutants affecting human and wildlife health (Sun et al., in the living organisms, such as steroid receptors, steroid synthesis, distribution and excretion, hypothalamus-pituitary-gonad (HPG) axis, as well as indirect mechanisms including thyroid and growth hormone disruption (Rempel and Schlenk, 2008). Many of the * Corresponding author. Department of Natural Resources, Isfahan University of anthropogenic activities are potential sources of this group of Technology, 84156-83111 Isfahan, Iran. fi fi E-mail address: [email protected] (N. Gilannejad). chemicals, which nally nd their way to rivers and lakes and affect http://dx.doi.org/10.1016/j.chemosphere.2015.09.106 0045-6535/© 2015 Elsevier Ltd. All rights reserved. N. Gilannejad et al. / Chemosphere 144 (2016) 1342e1350 1343 aquatic organisms (Jeffries et al., 2010). was quantified using real-time PCR. Moreover, condition factor, The Zayandeh Roud River is the only permanent and the most gonadosomatic index and sex ratio were calculated and related to important river in the central Iran, which plays a key role in vtg expression. Gonad histology was also performed with the goal providing Isfahan and the adjacent provinces with drinking water. of detecting intersex specimens. Land use along this river includes agricultural fields, cattle farms, municipal waste water treatment plants and heavy industries such 2. Materials and methods as steel mill companies, power plants and oil refinery installations. Therefore, there is a high probability of contamination by EDCs. 2.1. Studied area and sampling points According to our knowledge, the information about the presence and in vivo consequences of EDCs in water ecosystems from Isfahan The Zayandeh Roud River, with a drainage area of 41,500 km2, province or generally in Iran is very scarce. The limited data avail- is the largest river in the central plateau of Iran. Sampling sites able for the Zayandeh Roud River, which are dedicated to were selected throughout the river having in mind a possible measuring some heavy metal and a series of compounds belonging gradient of anthropogenic impacts. Cheshmeh Dimeh (S1) (32310 to polycyclic aromatic hydrocarbons, affirm their existence in water N e 50130 E), and Khersoonak (S2) (32310 N e 50220 E), are and/or sediments (Nemati et al., 2009; Abdollahi, 2013). located upstream of the Zayandeh Roud Dam and were considered Since chemical monitoring of the aquatic environment alone is as uncontaminated sites. In these two sites, some extensive and restricted to identification of some of the EDCs, and the detection scatter crop and cattle production can be found. On the other methods are costly and complex, the use of biomarkers in sentinel hand, Chamgordan (S3) (32230 N e 51170 E) and Safaieh Bridge species has been considered an effective approach to assess the (S4) (32210 N e 51260 E) are situated downstream of the health status of the aquatic ecosystems (Frenzilli et al., 2008; Blazer Zayandeh Roud Dam and were considered a priori as possible et al., 2012; Gilroy et al., 2012). Obviously, aquatic species are more contaminated sites. S3 is a lagoon like reservoir which is formed exposed to EDCs than terrestrial animals (Damstra et al., 2002). by entering the river into an enclosed area and it receives effluents Among different aquatic species used for EDCs screening, fishes are from the nearby major steel mill plant as well as agricultural fields. the prominent model due to their particular features, the most S4 is located downstream of the sewage treatment plant dis- important of which is that the basic aspects of the function and charges from the Zarin Shahr City (population in 2012: 95,326) structure of the HPG axis are conserved in all vertebrates. There- (Fig. 1). During fish collections, dissolved oxygen, temperature, pH, fore, the results from their study can be extrapolated to other conductivity, and total dissolved solids (TDS) were measured on vertebrates including humans (Ankley et al., 2009). Petroleuciscus site using WTW model OXI 196 Hand-Held Dissolved Oxygen esfahani is a small cyprinid fish endemic to the Zayandeh Roud Meter and HANNA model HI98129 pH/Conductivity/TDS Tester. basin (Coad and Bogutskaya, 2010). This species is abundant along Additionally, biochemical oxygen demand (BOD), and chemical the river and is even available in locations with constant input of oxygen demand (COD) were determined in laboratory according to contaminants. All these features, together with its limited migra- Radojevic and Bashkin (2006) (Table 1). tion capacity, make this fish a suitable candidate for sentinel spe- cies in this ecosystem. Fish exposure to EDCs with estrogenic and anti-androgenic ac- 2.2. Fish sampling and analyzed parameters tivity can lead to several consequences (Rempel and Schlenk, 2008), including i) vitellogenin induction in juveniles or males (Blazer Fish were captured during the non-reproductive period et al., 2014; Duffy et al., 2014), ii) delay or absence of secondary (January to February, 2012) by seine net (8 m  1 m, 5 mm mesh sexual characteristics and sexual behavior in males (Goksøyr et al., size) in S1, S2 and S4, and by boat electrofishing in S3 because of 2003), iii) intersex condition (Puy-Azurmendi et al., 2013), iv) higher water depth in this site. After transporting the fish to the skewed sex ratio (Jobling et al., 1996), and in extreme cases v) laboratory, on one side, 10 individuals were exposed to 17b- complete extinction of the population (Kidd et al., 2007). Vitello- estradiol (E2)(2mg/L) for 24 h (Bowman et al., 2000) with the genin (Vtg) is a female-specific precursor protein for egg yolk, purpose of increasing the chance of cloning the vtg cDNA. At the which is normally synthesized in female hepatocytes in response to end of this period, male and female liver samples were preserved ® endogenous estrogens (Goksøyr et al., 2003). Males also possess the in RNAlater (Invitrogen Life Technologies) separately. On the vitellogenin genes (vtg), which are silenced but can be induced other side, on fish arrival to the laboratory, the rest of the fish from when exposed to xenoestrogens (Hutchinson et al., 2005) and all the sampling sites were kept in aerated tanks with water from antiandrogens (Bhatia et al., 2014). Actually, vitellogenin induction the corresponding sampling site until processed (around 1 h). Fish in males is the strongest biomarker for fish exposure to estrogenic were anaesthetized by immersion in 100 ppm MS222. Length compounds

Load more

  9

Copy Link