Endocrine disruptor responses in African sharptooth catfish (Clarias gariepinus) exposed to di-(2-ethylhexyl)-phthalate.

In the present study, we have investigated the endocrine disruptive effects of waterborne di-(2-ethylhexyl) phthalate (DEHP: 0 (control), 10, 100, 200 and 400 µg/L) on juvenile Clarias gariepinus by analyzing transcript patterns for hepatic vitellogenin (vtg), estrogen receptor-α (er-α), aromatase (cyp19a1b) and peroxisome proliferator activated receptor-α (ppar-α) using quantitative real-time PCR after 3, 7 and 14 days exposure period. In addition, we analyzed CYP19 and PPAR protein levels using enzyme-linked immunosorbent assay (ELISA), while cellular testosterone (T) and 17ß-estradiol (E2) levels were measured using enzyme immune assay (EIA). Tissue burden of DEHP was measured in the liver using gas chromatography-mass spectroscopy (GC-MS). We observed apparent concentration- and time-dependent increases of vtg, er-α, cyp19a1b transcripts, E2 and T levels after exposure to DEHP. A biphasic pattern of effect was observed for ppar-α, showing a concentration-dependent increase that peaked at 100 µg/L and thereafter, an apparent concentration-dependent decrease at 200 and 400 µg/L at all exposure days. Given that the post-differentiation changes of gonads in C. gariepinus corresponded with the 14-day sampling period, we separated the analyzed data into sexes after histological examination of the gonads, showing that females responded stronger, compared to males, to DEHP exposure at all exposure concentrations. Oocyte atresia, intersex (ovotestis) and karyoplasmic clumping were observed in females while male fish showed distortion and degeneration of seminiferous tubules and condensation of tubular cells in the 400 µg/L exposure group after 14 days. Corresponding canonical analysis (CCA) of all analyzed variables revealed a positive relationship between analyzed biological variables with increasing DEHP concentrations. Overall, molecular, biochemical and physiological responses presented in the present study indicate that exposure of C. gariepinus to waterborne DEHP produced endocrine disruptive responses with potential consequences for overt reproduction, development, physiology and general health of fish populations inhabiting phthalate contaminated aquatic environments. These responses represent valuable and effective biomarkers of exposure and effects, that can be adopted for screening the presence of EDCs in Nigeria and other developing countries.

Endocrine-disruptor molecular responses, occurrence of intersex and gonado-histopathological changes in tilapia species from a tropical freshwater dam (Awba Dam) in Ibadan, Nigeria.

In the present study, the occurrence of endocrine disruptive responses in Tilapia species from Awba Dam has been investigated, and compared to a reference site (Modete Dam). The Awba Dam is a recipient of effluents from University of Ibadan (Nigeria) and several other anthropogenic sources. A total of 132 Tilapia species (Sarotherodon malenotheron (n=57 and 32, males and females, respectively) and Tilapia guineensis (n=23 and 20, males and females, respectively)) were collected from June to September 2014. At the reference site, samples of adult male and female S. melanotheron (48 males and 47 females) and T. guineensis (84 males and 27 females) were collected. Gonads were morphologically and histologically examined and gonadosomatic index (GSI) was calculated. Hepatic mRNA transcriptions of vitellogenin (Vtg) and zona radiata protein (Zrp) genes were analyzed using validated RT-qPCR. Significant increase in Vtg and Zrp transcripts were observed in male tilapias from Awba Dam, compared to males from the reference site. In addition, male tilapias from Awba Dam produced significantly higher Vtg and Zrp mRNA, compared to females in June and July. However, at the natural peak spawning period in August and September, females produced, significantly higher Vtg and Zrp mRNA, compared to males. Fish gonads revealed varying incidence of intersex with a striking presence of two (2) pairs of testes and a pair of ovary in S. melanotheron from Awba Dam. The entire fish population examined at Awba Dam showed a high prevalence of intersex (34.8%), involving phenotypic males and females of both species. Analysis of sediment contaminant levels revealed that As, Cd, Pb, Hg and Ni (heavy metals), monobutyltin cation, 4-iso-nonyphenol and PCB congeners (138, 153 and 180) were significantly higher in Awba Dam, compared to the reference site. Principal component analysis (PCA) showed that fish variables were positively correlated with sediment contaminant burden at Awba Dam, indicating that the observed endocrine disruptive responses are associated with contaminant concentrations. Overall, the occurrence of intersex and elevated expressions of Vtg and Zrp in male fish, suggest that the measured contaminants were eliciting severe endocrine disruptive effects in Awba Dam biota, which is an important source of domestic water supply and fisheries for the University of Ibadan community.

Occurrence, Species, and Organ Differences in Bioaccumulation Patterns of Phthalate Esters in Municipal Domestic Water Supply Lakes in Ibadan, Nigeria.

The occurrence of phthalate esters (PE) was examined in biota, ambient water, and sediments of two man-made lakes (Asejire and Eleyele) in southwestern Nigeria. Five fish species (Tilapia zillii, Hepsetus odoe, Parachanna obscura, Chrysichthys nigrodigitatus, and Mormyrus rume) were analyzed for PE levels and used for calculating bioconcentration factors (BCF) and biota-sediment accumulation factors (BSAF). In addition, measured PE levels were thereafter used to calculate the phthalate pollution index (PPI) in biota and the environment. At both lakes, all sampled species had k-factor > 1, showing apparently normal growth and health condition. Higher PE levels were found in sediments compared with water at both lakes, with a pattern showing that di(2-ethylhexyl) phthalate (DEHP) was predominant PE. While there were no unique patterns of PE concentrations in both lakes, differences were observed in organ concentration patterns that were evident at both lakes. For T. zillii, the BSAF was higher for dibutyl phthalate (DBP) compared to diethyl phthalate (DEP) and lowest for DEHP. The concentration pattern demonstrated that DBP concentrated more in gills (BCF: 6.7), while DEHP concentrated more in liver (BCF: 15.2) of T. zillii at Asejire. At Eleyele, T. zillii liver and gills concentrated less DEP and DEHP. The PPI value was significantly higher in sediment with respective values of 0.27 and 0.44 at Asejire and Eleyele lakes compared with water with respective values of 0.1 and 0.18 at Asejire and Eleyele lakes. Overall, our findings suggest a broader environmental and human health implication of high PE levels in these lakes, since they provide vast water volumes that are used for municipal domestic water supply. Further, these lakes support intense artisanal fisheries, representing significant sources of aquatic food resources for neighboring communities.