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Effect of Indigo Dye Effluent on the Growth, Biomass Production and Phenotypic Plasticity of Scenedesmus Quadricauda (Chlorococcales)

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Effect of Indigo Dye Effluent on the Growth, Biomass Production and Phenotypic Plasticity of Scenedesmus Quadricauda (Chlorococcales) Anais da Academia Brasileira de Ciências (2014) 86(1): 419-428 (Annals of the Brazilian Academy of Sciences) Printed version ISSN 0001-3765 / Online version ISSN 1678-2690 http://dx.doi.org/10.1590/0001-3765201420130225 www.scielo.br/aabc Effect of indigo dye effluent on the growth, biomass production and phenotypic plasticity of Scenedesmus quadricauda (Chlorococcales) MATHIAS A. CHIA1 and RILWAN I. MUSA2 1Laboratório de Cianobactérias, Escola Superior de Agricultura Luiz de Queiroz, Universidade de São Paulo, Av. Pádua Dias, 11, 13418-900 Piracicaba, SP, Brasil 2Department of Biological Sciences, Ahmadu Bello University, Zaria, Postal Code 810001, Nigeria Manuscript received on June 26, 2013; accepted for publication on October 14, 2013 ABSTRACT The effect of indigo dye effluent on the freshwater microalga Scenedesmus quadricauda ABU12 was investigated under controlled laboratory conditions. The microalga was exposed to different concentrations of the effluent obtained by diluting the dye effluent from 100 to 175 times in bold basal medium (BBM). The growth rate of the microalga decreased as indigo dye effluent concentration increased (p <0.05). The EC50 was found to be 166 dilution factor of the effluent. Chlorophyll a, cell density and dry weight production as biomarkers were negatively affected by high indigo dye effluent concentration, their levels were higher at low effluent concentrations (p <0.05). Changes in coenobia size significantly correlated with the dye effluent concentration. A shift from large to small coenobia with increasing indigo dye effluent concentration was obtained. We conclude that even at low concentrations; effluents from textile industrial processes that use indigo dye are capable of significantly reducing the growth and biomass production, in addition to altering the morphological characteristics of the freshwater microalga S. quadricauda. The systematic reduction in the number of cells per coenobium observed in this study further confirms that environmental stress affects coenobium structure in the genus Scenedesmus, which means it can be considered an important biomarker for toxicity testing. Key words: freshwater, microalgae, morphological variation, pollution, textile waste. INTRODUCTION treated, conventional waste treatment techniques Textile industrial processes consume large amounts do not completely decolorize and detoxify the dye of water and use toxic products. They contribute effluents (Puvaneswari et al. 2006). to the degradation of the environment by releasing Indigo dye is part of the numerous marketed untreated dye effluents into neighboring aquatic organic colorants used for coloration of textiles, ecosystems in the form of sludge. The final paper, leather, plastic and for specialized disposal of this sludge remains a challenge, and applications such as food, drug, cosmetic and its ecotoxicological assessment is important for photochemical productions (Zollinger 1987, minimizing its environmental impacts. Even when Novotny et al. 2006). Textile effluents containing indigo dye and other dye types make water toxic Correspondence to: Mathias Ahii Chia E-mail: [email protected] (Robinson et al. 2001) and unfit for human and An Acad Bras Cienc (2014) 86 (1) 420 MATHIAS A. CHIA and RILWAN I. MUSA animal consumption, and cause imbalance within the physiological state. Under environmental stress, different aquatic ecosystem food chains (Rocha it may fail to produce such colonies (van Den Hoek 1992). In recent years various researchers have et al. 1995, Lurling and van Donk 1999, Lombardi identified mutagenic effects of textile samples and et al. 2007). These changes in phenotypic plasticity waste water of the textile industry (Knasmüller et are considered a response to environmental stress al. 1993, Jäger 1998, Mathur et al. 2012). Their (Lurling and Beekman 2002, Peña-Castro et al. 2004). findings show that dyes used for textile finishing To the best of our knowledge, nothing is are mainly responsible for the mutagenic effects known about the effect of indigo dye effluents on observed. Although information on textile dye freshwater microalgae. Due to the extent to which toxicity to microalgae remain scanty, Rannug et the dye is extensively used today in the textile and al. (1992), Mathur et al. (2005a, b), and Mathur allied industries, understanding the effects of the and Bhatnagar (2007) were able to show that indigo dye effluents is very important and requires indigo dye had mutagenic effect on the bacterium investigations. Detection and quantification of the Salmonella typhimurium. morphological response is easy and inexpensive, The effluents of textile dyeing and printing and thus very relevant in environmental monitoring industries are complex in nature, rich in dissolved (Lombardi et al. 2007). This study was aimed at and suspended solids, organic compounds, heavy determining the effect of indigo dye effluent on metals and have high pH. The composition of the growth, biomass production and phenotypic textile effluents determines the extent to which plasticity of S. quadricauda. they pollute rivers (Srivastava et al. 1994), affect MATERIALS AND METHODS soil characteristics (Raj et al. 1997), and affect plants (Khandelwal 1996, Raj et al. 1997). Hence, The Scenedesmus quadricauda ABU12 strain used we hypothesize that the complex nature of indigo in this study was isolated from a freshwater pond in dye effluent would be responsible for the overall Zaria, Nigeria. It was maintained in the laboratory toxicity observed and not just the dye component with Bold Basal Medium (BBM) at 23+2°C alone. This is because the presence of metals like and under continuous illumination at 150 uml cadmium, chromium and copper in textile effluents photons m-2s-1 using cool white fluorescent lamps. can exhibit various interactive toxic effects on Culture media sterilization was performed through aquatic biota (Mathur et al. 2012). The final toxic autoclaving at 121°C for 15 min, and the media effect of the indigo dye effluent may be synergistic, were left to stand and equilibrate for 24 h before additive or antagonistic as a function of the different use. Sterile techniques were maintained throughout effluent components that make up the effluent and the experimental period. not the dye in isolation. Dye effluent samples were collected from local Scenedesmus chosen for this study is cosmo- textile industries located in Zaria, Nigeria. In order politan and can be found in a wide array of aquatic to confirm the identity of the dye used in the local ecosystems ranging from oligo-, meso-, eutrophic dye industries, we purchased raw indigo dye from and metal polluted waters. Scenedesmus quadricauda Emmybiz Dye and Chemicals International Limited, forms miniature colonies termed autocolonies or Sabon Gari, Zaria, Nigeria. This dye company coenobia, in which groups of 4 to 8 cells are connected supplies the local textile industries in Zaria with together in a row. The morphological form i.e. the indigo dye. The identification of the raw dye and the number of cells per coenobium of Scenedesmus dye in the effluent were done using spectroscopic varies depending on environmental conditions and methods by observing its absorption at different An Acad Bras Cienc (2014) 86 (1) THE TOXICITY OF INDIGO DYE EFFLUENT TO GREEN ALGAE 421 wavelengths with the aid of a SpectrumLab 7525 TABLE I UV-VIS spectrophotometer (B. Bran Scientific The physicochemical characteristics of the Indigo dye effluents used in the present study. and Instrument Company, England). In addition, Fourier Transform Infrared Analysis (FTIR) was Physicochemical parameters Value used to determine the different functional groups Nitrate nitrogen(mg L-1) 0.001 -1 of the dye as a way of confirming its identification. Phosphate phosphorous(mg L ) 50.500 Dissolved oxygen(mg L-1) 19.400 About 1 mg was weighed in a small agate mortar Chemical oxygen demand(mg L-1) 24.000 with a drop of nujol and Kbr. The mull was then Electrical conductivity 19.710 pressed between the flat plates of sodium chloride. Temperature (° C) 28.200 Before running a scan on each sample type, a pH 6.700 Biological oxygen demand(mg L-1) 0.000 background scan was carried out. All scanning was -1 -1 Cu(mg L ) 0.054 done from 4000 to 625 cm (Williams and Fleming Cr(mg L-1) 0.183 2005) using a Shimadzu FT-IR Model 8400s Cd(mg L-1) 0.003 spectrophotometer (Shimadzu, Japan). The effluent was characterized for physical made to a factor range of 100 to 175 times. The and chemical properties. pH and electrical conduc- control treatment with no dye effluent addition tivity levels were determined using HANNA pH/ was represented as normal BBM. The microalga EC/Temp meter model 210, and turbidity was was acclimated to the BBM using semi-continuous determined spectrophotometrically. Biochemical batch system. When the microalga was at mid oxygen demand, dissolved oxygen and chemical exponential growth phase, dilutions were made oxygen demand were determined using the Azide by re-inoculating at a 10% v/v until a steady state Modification of the Winkler method as described physiological growth status was obtained. This is in APHA (1998). Nitrate nitrogen, phosphate the stage where the growth rate was statistically the phosphorus and total sulphate were determined same and reflecting the growth condition to which using the spectrophotometric procedures described S. quadricauda was exposed. It is this exponential in APHA (1998). Cadmium, chromium and copper phase growing S. quadricauda that was exposed
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