Variability of Bile Baseline Excitation-emission Fluorescence of Two Tropical Freshwater Fish Species.

The quantification of pollutant metabolites in fish bile is an efficient approach to xenobiotic pollution monitoring in freshwaters since these measurements directly address exposure. Fluorescence excitation-emission matrix spectroscopy (EEMS) has demonstrated to be a highly specific and cost-effective technique for polycyclic aromatic hydrocarbon (PAH) and PAH-metabolite identification and quantification. EEMS ability to quantify these compounds strongly depends on the intensity and variability of the bile baseline fluorescence (BBF). We found large differences in BBF among Aequidens metae (AME) individuals and of these with Piaractus orinoquensis (PIO). Moreover, BBF was large enough that solvent dilutions of over 1:400 were needed to avoid inner filter effects. We used parallel factor analysis (PARAFAC) to model the intra- and inter-species BBF variability. PARAFAC successfully decomposed the EEMS set into three fluorophores present in all samples, although in concentrations spreading over ~ 3 orders of magnitude. One of the factors was identified as tryptophan. Tryptophan and Factor 2 were covariant and much more abundant in AME than in PIO, while Factor 3 was ~ 6 times more abundant in PIO than in AME. Also, tryptophan was ~ 10x more abundant in AME specimens immediately caught in rivers than in their laboratory-adapted peers. The PARAFAC decomposition effectiveness was confirmed by the positive proportionality of scores to dilution ratios. A large inner filter indicates that Factor 2 is as strong a light absorber as tryptophan. Our results stress the need to include bile matrix variable components for the detection and quantification of pollutant metabolites using PARAFAC.

Liver biomarkers response of the neotropical fish Aequidens metae to environmental stressors associated with the oil industry.

The Acacias River in Colombia receives large volumes of industrial effluents mostly derived from the oil industry. To contribute to the study of the possible effects of industrial wastewaters on the aquatic environment and particularly on fish populations, a native neotropical fish, Aequidens metae was used as a sentinel species. Wild specimens of A. metae were caught at three different places of the Acacias River taking as reference the point of discharge of an oil industry effluent; upstream, downstream, and at the vicinity of the discharge pipe. A fourth sampling site was chosen as a reference site away from urban settlements. Samplings were performed twice, during the rainy and dry seasons. After anesthesia animals were weighted and measured, and humanely sacrificed. Livers were extracted, frozen on site and transported to the laboratory. Condition indices were calculated. Total protein content and the detoxification 7-ethoxyresorufin-O-deethylase (EROD) enzyme activity were estimated. Histopathological alterations were also evaluated. Water quality was estimated through the measurement of several variables. Results obtained evidenced that the highest induction in EROD activity and the strongest histological alterations in liver of the monitored fish appeared during the dry seasons at the discharge site and downstream to this point.

Biochemical responses and proximate analysis of Piaractus brachypomus (Pisces: Characidae) exposed to phenanthrene.

Polycyclic Aromatic Hydrocarbons (PAHs) are complex compounds generated from industrial and anthropogenic activities, although natural phenomena are also responsible for their presence in the environment. The aim of this study was to evaluate the effect of phenanthrene on proximate composition and antioxidant activity in Piaractus brachypomus during a subacute exposure. Fish were exposed intraperitoneally to phenanthrene (0.1, 1.0, and 10 µg g-1) and a solvent control (0 µg g-1). Muscle tissue, liver and bile were collected at 0 h, 11 days and 21 days. In dorsal muscle, parameters such as dry matter, protein, crude fat, and pH did not show alterations with exposure to phenanthrene (p ≥ 0.05). At 11 days, a significant increase of hepatic lipid peroxidation was observed in fish exposed to 10 µg g-1. Additionally, a dose-dependent response was observed in the muscle, although no significant differences were observed in the activity of catalase and lipid peroxidation. Phenanthrene metabolites in bile were analyzed by fixed fluorescence at 260/380 nm (excitation/emission) wavelengths and levels indicated a dose-dependent response. Likewise, bioaccumulation of phenanthrene in bile was observed for 21 days, suggesting slow metabolism of this xenobiotic at 10 µg g-1. This study provides important information on the oxidative effects generated by phenanthrene exposure for 21 days in P. brachypomus, generating fundamental information on adverse effects induced by PAHs on the most important Colombian native fish species. These data contribute to the development of additional research in environments contaminated with PAHs and can be applied to monitor areas contaminated with oil spills.

Physiological and enzymatic responses of Chlorella vulgaris exposed to produced water and its potential for bioremediation.

In South America, Colombia is known as an important oil-producing country. However, the environmental impact of crude oil industry has not been studied deeply and few studies have been carried out for evaluating responses of algae and its adaptation under specific conditions. Enzymatic and physiological effects in Chlorella vulgaris and its potential for bioremediation after exposure to produced water (PW) were assessed using different PW concentrations (0, 25, 50, 75 and 100%) and crude oil. Variables such as cell density, growth rate (µ), percentage of growth inhibition (% I), chlorophyll a and b and cell diameter were evaluated during 5 days. Furthermore, enzymatic biomarkers such as superoxide dismutase (SOD) and catalase (CAT) were also measured. Results showed that the treatment with 100% PW had the highest cell density and µ; similarly, 25% PW treatment had a similar behaviour, being these two treatments with the highest growth. A dose-dependent response was seen for chlorophyll a and b and cell diameter, showing significant differences between treatments and the control. Different levels of SOD and CAT were observed in algae exposed to PW. At 24 h, an increase in SOD and CAT activity was observed, probably due to effects caused by xenobiotics. After 72 h, a decrease in the activity of both enzymes was observed. The results evidenced that C. vulgaris can adapt easily to PW, showing an increase on its growth and stabilisation in its antioxidant activity. Additionally, cell diameter results and decrease of hydrocarbons and phenols show the potential of these algae to degrade xenobiotics from PW.