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<article article-type="research-article" dtd-version="1.0" specific-use="sps-1.8" xml:lang="en" xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink">
	<front>
		<journal-meta>
			<journal-id journal-id-type="publisher-id">rica</journal-id>
			<journal-title-group>
				<journal-title>Revista internacional de contaminación ambiental</journal-title>
				<abbrev-journal-title abbrev-type="publisher">Rev. Int. Contam.
					Ambient</abbrev-journal-title>
			</journal-title-group>
			<issn pub-type="ppub">0188-4999</issn>
			<publisher>
				<publisher-name>Universidad Nacional Autónoma de México, Centro de Ciencias de la Atmósfera</publisher-name>
			</publisher>
		</journal-meta>
		<article-meta>
			<article-id pub-id-type="doi">10.20937/RICA.53375</article-id>
			<article-id pub-id-type="publisher-id">00014</article-id>
			<article-categories>
				<subj-group subj-group-type="heading">
					<subject>Artículos</subject>
				</subj-group>
			</article-categories>
			<title-group>
				<article-title>EFFICIENCY OF EFFLUENT TREATMENT OF MEATPACKING AND TEXTILE PLANTS,
					IN PHYSICAL, CHEMICAL AND TOXICOLOGICAL TERMS</article-title>
				<trans-title-group xml:lang="es">
					<trans-title>EFICIENCIA DEL TRATAMIENTO DE EFLUENTES DE PLANTAS DE CARNE Y
						PLANTAS TEXTILES, EN TÉRMINOS FÍSICOS, QUÍMICOS Y
						TOXICOLÓGICOS</trans-title>
				</trans-title-group>
			</title-group>
			<contrib-group>
				<contrib contrib-type="author">
					<name>
						<surname>Fontana</surname>
						<given-names>Michelli</given-names>
					</name>
					<xref ref-type="aff" rid="aff1"><sup>1</sup></xref>
				</contrib>
				<contrib contrib-type="author">
					<name>
						<surname>Lopes Turino</surname>
						<given-names>Rafael</given-names>
					</name>
					<xref ref-type="aff" rid="aff1"><sup>1</sup></xref>
				</contrib>
				<contrib contrib-type="author">
					<name>
						<surname>Benedetti Tonial</surname>
						<given-names>Ivane</given-names>
					</name>
					<xref ref-type="aff" rid="aff1"><sup>1</sup></xref>
				</contrib>
				<contrib contrib-type="author">
					<name>
						<surname>Sauer Pokrywiecki</surname>
						<given-names>Ticiane</given-names>
					</name>
					<xref ref-type="aff" rid="aff1"><sup>1</sup></xref>
				</contrib>
				<contrib contrib-type="author">
					<name>
						<surname>Düsman</surname>
						<given-names>Elisângela</given-names>
					</name>
					<xref ref-type="aff" rid="aff1b"><sup>1</sup></xref>
					<xref ref-type="corresp" rid="c1">*</xref>
				</contrib>
				</contrib-group>
				<aff id="aff1">
					<label>1</label>
					<institution content-type="original">Universidade Tecnológica Federal do Paraná,
						Departamento Acadêmico de Química e Biología, Linha Santa Bárbara s/n, CEP
						85601-970 - Caixa Postal 135 - Francisco Beltrão - PR - Brasil</institution>
					<institution content-type="normalized">Universidade Tecnológica Federal do
						Paraná</institution>
					<institution content-type="orgdiv1">Universidade Tecnológica Federal do
						Paraná</institution>
					<institution content-type="orgname">Departamento Acadêmico de Química e
						Biología</institution>
					<addr-line>
						<named-content content-type="city">Francisco Beltrão</named-content>
					</addr-line>
					<country country="BR">Brazil</country>
				</aff>
				<aff id="aff1b">
					<label>1</label>
					<institution content-type="original">Universidade Tecnológica Federal do Paraná,
						Departamento Acadêmico de Química e Biología, Linha Santa Bárbara s/n, CEP
						85601-970 - Caixa Postal 135 - Francisco Beltrão - PR - Brasil</institution>
					<institution content-type="normalized">Universidade Tecnológica Federal do
						Paraná</institution>
					<institution content-type="orgdiv1">Universidade Tecnológica Federal do
						Paraná</institution>
					<institution content-type="orgname">Departamento Acadêmico de Química e
						Biología</institution>
					<addr-line>
						<named-content content-type="city">Francisco Beltrão</named-content>
					</addr-line>
					<country country="BR">Brazil</country>
					<email>edusman@utfpr.edu.br</email>
				</aff>
			<author-notes>
				<corresp id="c1">
					<label>*</label>Corresponding author: <email>edusman@utfpr.edu.br</email>
				</corresp>
			</author-notes>
			<!--<pub-date date-type="pub" publication-format="electronic">
				<day>04</day>
				<month>05</month>
				<year>2021</year>
			</pub-date>
			<pub-date date-type="collection" publication-format="electronic">-->
				<pub-date pub-type="epub-ppub">
				<month>05</month>
				<year>2020</year>
			</pub-date>
			<volume>36</volume>
			<issue>2</issue>
			<fpage>399</fpage>
			<lpage>411</lpage>
			<history>
				<date date-type="received">
					<day>01</day>
					<month>08</month>
					<year>2018</year>
				</date>
				<date date-type="accepted">
					<day>01</day>
					<month>08</month>
					<year>2019</year>
				</date>
			</history>
			<permissions>
				<license license-type="open-access"
					xlink:href="https://creativecommons.org/licenses/by-nc-nd/4.0/" xml:lang="en">
					<license-p>This is an open-access article distributed under the terms of the
						Creative Commons Attribution License</license-p>
				</license>
			</permissions>
			<abstract>
				<title>ABSTRACT</title>
				<p>The disposal of untreated effluents directly affects the water quality of the
					receiving medium, which can cause damage to the aquatic ecosystem and to human
					health. Therefore, the objective of this study was to analyze the toxicity and
					the physical and chemical parameters of raw and treated effluents of a
					meatpacking and a textile plant, in order to evaluate the effectiveness of the
					treatment. Thus, cytotoxic tests with <italic>Allium cepa</italic> L. and the
					immobility/mortality test with <italic>Artemia salina</italic> L., besides the
					physical and chemical tests of pH, ammoniacal nitrogen, oxygen demand, total
					solids, chlorides and alkalinity were used. The immobility/mortality test with
						<italic>Artemia salina</italic> L. revealed the absence of dead organisms in
					the tested low concentrations of raw and treated meatpacking and textile
					effluents. However, the highest concentrations of raw meatpacking effluents (50
					% and 100 %) and treated meatpacking effluents (100 %) caused the mortality of
					organisms. Raw and treated textile effluents were statistically toxic only at
					the highest concentration evaluated (100 %). Furthermore, the raw and treated
					effluents of the meatpacking and textile plants were not cytotoxic in the test
					with <italic>Allium cepa</italic> L. The results of the physicochemical analyses
					proved the effectiveness of the treatment of the meatpacking effluents in terms
					of diminished pH, ammoniacal nitrogen, chemical oxygen demand, and total solids.
					And the raw effluents from the textile plant also exhibited a reduction in pH,
					chlorides, alkalinity and total solids. The treatments were effective for these
					effluents, but still had a toxic effect on <italic>A. salina</italic>. Thus, it
					is indicated that additional treatment should be carried out in the textile and
					meatpacking effluents, emphasizing the importance of adequate treatment systems
					to prevent environmental harm and public health problems.</p>
			</abstract>
			<trans-abstract xml:lang="es">
				<title>RESUMEN</title>
				<p>La liberación de efluentes no tratados afecta directamente la calidad del agua
					del medio receptor, pudiendo traer perjuicios al ecosistema acuático y también a
					la salud humana. Por lo tanto, el objetivo de este estudio fue analizar la
					toxicidad y los parámetros fisicoquímicos del efluente con y sin tratamiento de
					una planta textil y una empacadora de carne, a fin de evaluar la efectividad del
					tratamiento utilizado. Se utilizaron pruebas citotóxicas con <italic>Allium
						cepa</italic> L. y la prueba de inmovilidad/mortalidad con <italic>Artemia
						salin</italic>a L., además de las pruebas físicas y químicas de pH,
					nitrógeno amoniacal, demanda de oxígeno, sólidos totales, cloruros y
					alcalinidad. La prueba de inmovilidad/mortalidad con <italic>Artemia
						salina</italic> L. reveló la ausencia de organismos muertos en las bajas
					concentraciones probadas de los efluentes en bruto y tratados de la empacadora y
					la fábrica textil. Sin embargo, las concentraciones más altas de los efluentes
					no tratados de la empacadora de carne (50 y 100 %) y los efluentes tratados de
					dicha planta (100 %) causaron la mortalidad de organismos. Los efluentes de la
					fábrica textil tratados y sin tratamiento fueron estadísticamente tóxicos sólo
					en la mayor concentración evaluada (100 %). Además, los efluentes con y sin
					tratamiento de ambas plantas no fueron citotóxicos en la prueba con
						<italic>Allium cepa</italic> L. Los resultados de los análisis físicos y
					químicos demostraron la efectividad del tratamiento de los efluentes de la
					empacadora de carne en términos de pH disminuido, nitrógeno amoniacal, demanda
					química de oxígeno y sólidos totales. Y los efluentes sin tratar de la planta
					textil también mostraron una reducción en el pH, cloruros, alcalinidad y sólidos
					totales. Los tratamientos fueron efectivos para estos efluentes, pero aún así
					tuvieron un efecto tóxico sobre <italic>A. salina</italic>. Por lo tanto, está
					indicado realizar un tratamiento adicional en los efluentes de ambas plantas,
					enfatizando la importancia de contar con sistemas de tratamiento adecuados para
					prevenir daños ambientales y problemas de salud pública.</p>
			</trans-abstract>
			<kwd-group xml:lang="en">
				<title>Key words:</title>
				<kwd><italic><italic>Allium cepa</italic></italic> L.</kwd>
				<kwd><italic>Artemia salina</italic> L.</kwd>
				<kwd>meatpacking effluent</kwd>
				<kwd>textile effluent</kwd>
				<kwd>toxicity</kwd>
			</kwd-group>
			<kwd-group xml:lang="es">
				<title>Palabras clave:</title>
				<kwd><italic><italic>Allium cepa</italic></italic> L.</kwd>
				<kwd><italic>Artemia salina</italic> L.</kwd>
				<kwd>efluente de empacado de carne</kwd>
				<kwd>efluente textil</kwd>
				<kwd>toxicidad</kwd>
			</kwd-group>
			<counts>
				<fig-count count="1"/>
				<table-count count="5"/>
				<equation-count count="0"/>
				<ref-count count="47"/>
				<page-count count="13"/>
			</counts>
		</article-meta>
	</front>
	<body>
		<sec sec-type="intro">
			<title>INTRODUCTION</title>
			<p>One of the main sources of water resource contamination is the disposal of domestic
				and industrial effluents. These effluents compromise the quality of the entire
				aquatic ecosystem (<xref ref-type="bibr" rid="B12">Earnhart 2013</xref>, <xref
					ref-type="bibr" rid="B21">Kamali et al. 2019</xref>). In particular, liquid
				industrial effluents contain organic matter and inorganic chemical compounds that
				are hazardous and toxic to any kind of life, whether aquatic or not. The inability
				of industries to meet the effluent discharge standards and regulations can
				negatively affect the environment (<xref ref-type="bibr" rid="B33">Prabakar et al.
					2018</xref>).</p>
			<p>Given the context of industrial activities and based on the reports of <xref
					ref-type="bibr" rid="B27">Mathur et al. (2007)</xref>, <xref ref-type="bibr"
					rid="B43">Sharma et al. (2007)</xref> and <xref ref-type="bibr" rid="B30">Noreen
					et al. (2017)</xref>, the textile sector is considered the most polluting
				sector. According to <xref ref-type="bibr" rid="B44">Silva et al. (2004),
				</xref>every year 12 % of synthetic dyes used in textile production are wasted and
				dumped into the environment, damaging the photosynthetic processes in water
				channels.</p>
			<p>Moreover, these effluents have a complex chemical composition, with large amounts of
				suspended solids, high chemical oxygen demand (COD) and biochemical oxygen demand
				(BOD), and high temperatures, acidity and/or alkalinity (<xref ref-type="bibr"
					rid="B24">Liu et al. 2007</xref>, <xref ref-type="bibr" rid="B30">Noreen et al.
					2017</xref>, <xref ref-type="bibr" rid="B11">Costa et al. 2018</xref>). They
				also contain anti-foaming agents and humectants, oils, metals, and other organic
				compounds used in the various stages of dyeing (<xref ref-type="bibr" rid="B44"
					>Silva et al. 2004</xref>, <xref ref-type="bibr" rid="B24">Liu et al.
					2007</xref>, <xref ref-type="bibr" rid="B27">Mathur et al. 2007</xref>, <xref
					ref-type="bibr" rid="B30">Noreen et al. 2017</xref>).</p>
			<p>In addition to the textile industry, the food industry has also contributed to the
				rising pollution of water resources. In recent years, the increasing population has
				led to a growing demand for meat and the enhancement of slaughter and meat
				industrialization techniques, which generate large volumes of wastewater with highly
				polluting characteristics (<xref ref-type="bibr" rid="B46">Sperling 2007</xref>,
					<xref ref-type="bibr" rid="B26">Maldaner 2008</xref>).</p>
			<p>Regarding this type of industry, the slaughter of poultry produces on average 25 to
				50 liters of water per head, bovine slaughter generates 2500 liters per head, and
				pig slaughter requires 1200 liters per head (<xref ref-type="bibr" rid="B26"
					>Maldaner 2008</xref>).</p>
			<p>The effluents of the meat product industry contain large amounts of animal remains,
				fragments of viscera and tissue, blood, and fat. This waste leads to high levels of
				suspended solids, organic matter, organic and ammoniacal nitrogen, phosphorus, COD,
				and BOD, in addition to detergents and disinfectants used for plant cleaning
				operations, contributing significantly to the pollution of water sources (<xref
					ref-type="bibr" rid="B40">Scarassati et al. 2003</xref>, <xref ref-type="bibr"
					rid="B28">Mees 2006</xref>, <xref ref-type="bibr" rid="B26">Maldaner
				2008</xref>, <xref ref-type="bibr" rid="B29">Nogueira 2010</xref>). Furthermore, and
				due to its constitution, the effluents are highly fetid and quickly decompose (<xref
					ref-type="bibr" rid="B29">Nogueira 2010</xref>), possibly triggering the process
				of eutrophication when released directly into the water body (<xref ref-type="bibr"
					rid="B28">Mees 2006</xref>).</p>
			<p>Many meatpacking plants launch the effluents directly into watercourses. These
				effluents may be diluted, according to the size of the river and volume of water,
				which can minimize the environmental impact (<xref ref-type="bibr" rid="B26"
					>Maldaner 2008</xref>); however, this wastewater must be appropriately treated
				before being released (<xref ref-type="bibr" rid="B40">Scarassati et al.
				2003</xref>).</p>
			<p>The reality of industrial effluent treatment is highly diverse; although it is
				theoretically possible to treat effluents to meet any legal standard and permit
				their release into watercourses, there are several limiting factors in the everyday
				activities of the plants, such as high costs and technological deficiencies (<xref
					ref-type="bibr" rid="B41">Schoenhals 2006</xref>).</p>
			<p>Facing water pollution caused by the lack of appropriate treatment for effluents, the
				necessity to ensure effective effluent treatment emerges. These considerations
				reveal the need to enhance and monitor the treatment of effluents, and establish
				more efficient, cost-effective methods. According to <xref ref-type="bibr" rid="B5"
					>Bhat et al. (2019)</xref> efficient biomonitoring of effluents for its
				treatments using eco-friendly technologies could be beneficial to reduce the
				pollution of water ecosystem.</p>
			<p>The most widely used method to assess wastewater pollution levels and consequently
				predict environmental risks are physical and chemical analyses, which identify and
				quantify the concentrations of toxic substances, and ecotoxicological tests, which
				verify the effect of these substances on living organisms (<xref ref-type="bibr"
					rid="B10">Costa et al. 2008</xref>). Toxicity tests are important to evaluate
				the bio-efficiency of the treatments applied to effluents, considering that
				pollutant degradation products can be more toxic than the compounds of origin (<xref
					ref-type="bibr" rid="B6">Bilal et al. 2016a</xref>, <xref ref-type="bibr"
					rid="B7">b</xref>).</p>
			<p>It is recommended that the toxic effect of a sample be evaluated by more than one
				species, preferably belonging to different trophic levels of the food chain (<xref
					ref-type="bibr" rid="B25">Luíz et al. 2012</xref>), such as the
				immobility/mortality test with <italic>Artemia salina</italic> L. and the micro
				shellfish and cytotoxicity test with the vegetable <italic>Allium cepa</italic> L.
				The test with <italic>A. salina</italic> is fast and inexpensive, it is easy to
				handle and maintain, and is sufficiently reliable (<xref ref-type="bibr" rid="B17"
					>Hirota et al. 2012</xref>, <xref ref-type="bibr" rid="B22">Libralato et al.
					2016</xref>, <xref ref-type="bibr" rid="B30">Noreen et al. 2017</xref>). The
				test with <italic>A. cepa</italic> is also inexpensive and reliable, and is in
				agreement with other tests for genotoxicity, aiding studies to prevent damage to
				human health (<xref ref-type="bibr" rid="B19">Iqbal et al. 2019</xref>).</p>
			<p>Thus, the aim of the present study was to analyze the toxicity of raw and treated
				effluents of a meatpacking and a textile plant in the southwest of the state of
				Paraná, using the immobility/mortality test with <italic>Artemia salina</italic> L.
				as bioindicator, and the cytotoxicity test with <italic>Allium cepa</italic> L. to
				verify whether the treatment applied by both plants was efficient and resulted in
				diminished effluent toxicity. Moreover, we sought to establish the physical and
				chemical parameters of the raw and treated effluents to assess the effectiveness of
				the treatments. Therefore, this study can provide information about the
				effectiveness of the conventional system of treating effluents currently used in the
				textile and meatpacking plants, to prevent environmental harm and public health
				problems.</p>
		</sec>
		<sec sec-type="materials|methods">
			<title>MATERIALS AND METHODS</title>
			<sec>
				<title>Sample collection</title>
				<p>The liquid effluents from two industries located in the southwestern region of
					the Paraná state were selected for analysis, being one a meatpacking plant and
					the other a textile facility. The liquid effluents from both industries were
					evaluated before and after the treatment performed by each one. One collection
					of the samples was performed with a stainless-steel bucket previously sanitized
					and sterilized with a capacity of ten liters. Eight liters of liquid were
					collected from the meatpacking effluent, 4 L from the raw meatpacking effluent
					(RME) before the treatment, and 4 L from the treated meatpacking effluent (TME).
					The same procedure was used to collect samples of the liquid effluent from the
					textile industry: 4 L from the raw textile (RTE), and 4 L from the treated
					textile effluent (TTE). The samples were stored in resistant plastic vials
					chemically inert and tightly sealed, correctly cleaned, sterilized and labeled.
					The vials were maintained under refrigeration in thermal boxes for
					transportation to the Francisco Beltrão campus of the Federal University of
					Paraná (UTFPR-FB), where they were transferred to refrigerator and kept at 4 ºC
					until their analysis. With the exception of pH and temperature analyses, which
					were made in situ, the others were performed in the Water, Effluent and Biology
					Laboratory of UTFPR (<xref ref-type="bibr" rid="B2">APHA 1998</xref>).</p>
				<p>The industrial treatment of the meatpacking effluent was composed of a Parshall
					trough, whose function was to measure the inlet and outlet flows of the system
					effluent and to act as a quick mixer, together with a grid responsible for the
					removal of coarse solids. After this, the effluent passed through a grease box
					that allowed the removal of the oils and free greases by density difference,
					followed by a desander, whose objective was to separate the sand and other
					disposable debris from the liquid medium. Finally, the effluent advanced through
					three lagoons of anaerobic stabilization.</p>
				<p>The treatment of the textile effluent used by the industry was characterized by
					an equalization tank, a rotary sieve for the mechanical removal of coarse
					solids, coagulation/flocculation, pH correction, and the action of an aluminum
					sulfate coagulating agent, followed by biological treatment with mechanic
					aerators that oxygenate water.</p>
			</sec>
			<sec>
				<title>Physical and chemical analyses</title>
				<p>In accordance with the standards established in resolution 430 of the National
					Environmental Council (<xref ref-type="bibr" rid="B9">CONAMA 2011</xref>) and
					described in the standard methods of the American Public Health Association
						(<xref ref-type="bibr" rid="B2">APHA 1998</xref>), the physical and chemical
					characterization of the raw and treated effluents examined the following
					parameters: pH, temperature, chemical oxygen demand (COD), dissolved oxygen
					(DO), ammoniacal nitrogen, total solids, alkalinity and chloride. The tests of
					the raw and treated effluent samples from the meatpacking and textile industries
					were carried out by triplicate. Data were evaluated considering the average
					between the obtained results of each analysis. The Tukey test was used for
					statistical analysis (α = 0.05; n = 3).</p>
			</sec>
			<sec>
				<title><bold>Mortality/immobility test with <italic>
							<italic>Artemia salina</italic> L.</italic>
					</bold></title>
				<p>The acute toxicity test with <italic>Artemia salina</italic> L. was conducted
					according to the method proposed by <xref ref-type="bibr" rid="B15">Guerra
						(2001)</xref> and <xref ref-type="bibr" rid="B30">Noreen et al.
						(2017)</xref> with modifications. Initially, cysts of <italic>A.
						salina</italic>, which were obtained from an aquarium store, were incubated
					in synthetic sea salt solution (30 g/L), with light, and aerated at 25 ºC to
					induce eclosion.</p>
				<p>After hatching, 10 nauplii in instar 2-3 stages were transferred to test tubes
					containing 2 mL of the treatment samples, diluted in saline solution in the
					following concentrations: 100, 50, 25, 12.5, 6.2, and 3.1 %, to evaluate the
					effect of different effluent concentrations. A negative control group was
					prepared with 2 mL of saline solution (Co<sup>-</sup>). Four biological
					replicates were performed, containing 10 organisms in each replicate of each
					sample group. After 24 h of incubation at 25 ºC, the number of dead nauplii was
					determined by counting those that remained motionless for 20 s.</p>
				<p>Toxicity results were expressed as mortality percentage using the following
					formula: % toxicity = (average number of immobile individuals in biological
					replicates in each concentration)/(total mean number of individuals in
					biological replicates in each concentration) × 100. The mean (± standard
					deviation) number of dead organisms, calculated from the number of dead
					organisms in each biological replicate of each sample was compared using the
					Tukey test (α = 0.05; n = 4). </p>
			</sec>
			<sec>
				<title><bold>Cytotoxicity test with <italic>
							<italic>Allium cepa</italic> L.</italic>
					</bold></title>
				<p>The cytotoxicity test with <italic>Allium cepa</italic> L. was conducted
					according to the method proposed by <xref ref-type="bibr" rid="B13">Fiskesjö
						(1985)</xref>. The cytotoxicity of the samples was assessed using the
					meristematic cells of the root of <italic>Allium cepa</italic> L. as a test
					system, prepared with Feulgen reaction and stained with the Schiff reagent</p>
				<p>Onion bulbs were placed for four days in jars with water at room temperature,
					aerated and in the dark, to put down roots. Before each treatment, three roots
					were collected and fixed (methanol:acetic acid solution 3:1 v:v) to serve as
					control of the bulb itself (0 h). Then, the roots of these bulbs were placed in
					the samples of raw and treated effluents of the meatpacking and textile plants
					in 100 % concentration for 24 h. After the treatment time, three roots of each
					onion were removed and fixed (24 h). The remaining roots were washed, and the
					bulbs were placed again in filtered water to recover for 24 h, after which the
					roots were removed and fixed (48 h). In the negative control group, the onions
					remained in filtered water during the entire sampling time.</p>
				<p>Next, the roots were prepared in stages involving washing, hydrolysis with
					hydrochloric acid (HCl 1 N at 60 ºC for 10 min) and staining with Schiff’s
					reagent (for 45 min without light). The meristematic region of the roots was
					used to build the slides, prepared by maceration, and stained with acetic
					orcein. </p>
				<p>The slides were analyzed in a blind test under a light microscope with a 40×
					objective. Five bulbs were used for each control and treated group, and 1000
					cells were analyzed per bulb, differentiated according to the mitotic phases
					(interphase, prophase, metaphase, anaphase, telophase), totaling 5000 cells at
					the collection times of 0, 24, and 48 h. The mitotic index percentage (MI %) was
					calculated using the following formula: MI % = (number of dividing cells/total
					number of cells analyzed) × 100. The percentage of the phase index was
					calculated by the following formula: number of cells in each phase of the
					mitotic cell cycle/total number of cells analyzed × 100.</p>
				<p>The MIs and the phase index were statistically analyzed by the normality test and
					ANOVA, followed by the Tukey averages comparison test (α = 0.05, n = 5), using
					Action 6.2 software.</p>
			</sec>
		</sec>
		<sec sec-type="results|discussion">
			<title>RESULTS AND DISCUSSION</title>
			<p>Data from the toxicity test with the microcrustacean <italic>A. salina</italic>
					(<xref ref-type="table" rid="t1">Table I</xref>) showed an insignificant number
				of dead/immobile organisms in the low concentrations tested (3.1, 6.2, 12.5 and 25
				%) of the raw and treated meatpacking and textile effluents, when compared to the
				average mortality/immobility number of the negative control. However, the percentage
				of toxicity (<xref ref-type="table" rid="t2">Table II) </xref>varied from 7.5 to
				76.6 % and 5 to 92.5 % in the raw and treated meatpacking effluents, respectively,
				and from 7.5 to 100 % and from 0 to 91.6 % in the raw and treated textile effluents,
				respectively.</p>
			<p>
				<table-wrap id="t1">
					<label>TABLE I</label>
					<caption>
						<title>MEANS AND STANDARD DEVIATIONS OF THE NUMBER OF DEAD/IMMOBILE
								<italic>Artemia salina</italic> L. FOR THE FOUR BIOLOGICAL
							REPLICATES</title>
					</caption>
					<table frame="hsides" rules="groups">
						<colgroup>
							<col/>
							<col/>
							<col/>
							<col/>
							<col/>
						</colgroup>
						<tbody>
							<tr>
								<td align="center">Concentration</td>
								<td align="center">RME</td>
								<td align="center">TME</td>
								<td align="center">RTE</td>
								<td align="center">TTE</td>
							</tr>
							<tr>
								<td align="center">Co<sup>-</sup> (%)</td>
								<td align="center">0.25 ± 0.50a</td>
								<td align="center">0.25 ± 0.50a</td>
								<td align="center">0.25 ± 0.50a</td>
								<td align="center">0.25 ± 0.50a</td>
							</tr>
							<tr>
								<td align="center">3.1</td>
								<td align="center">2.00 ± 1.41abA</td>
								<td align="center">1.00 ± 1.41aA</td>
								<td align="center">0.50 ± 1.00aA</td>
								<td align="center">1.96 ± 2.62aA</td>
							</tr>
							<tr>
								<td align="center">6.2</td>
								<td align="center">0.75 ± 0.95abA</td>
								<td align="center">1.00 ± 1.41aA</td>
								<td align="center">0.75 ± 0.5aA</td>
								<td align="center">3.42 ± 4.75aA</td>
							</tr>
							<tr>
								<td align="center">12.5</td>
								<td align="center">0.91 ± 0.11abA</td>
								<td align="center">1.75 ± 1.50aA</td>
								<td align="center">1.25 ± 1.25aA</td>
								<td align="center">0.00 ± 0.0aA</td>
							</tr>
							<tr>
								<td align="center">25</td>
								<td align="center">0.75 ± 0.50abA</td>
								<td align="center">0.50 ± 0.58aA</td>
								<td align="center">0.75 ± 0.95aA</td>
								<td align="center">0.35 ± 0.71aA</td>
							</tr>
							<tr>
								<td align="center">50</td>
								<td align="center">3.20 ± 0.63bA</td>
								<td align="center">0.50 ± 0.58aA</td>
								<td align="center">2.25 ± 2.87aA</td>
								<td align="center">2.47 ± 1.33aA</td>
							</tr>
							<tr>
								<td align="center">100</td>
								<td align="center">7.66 ± 1.76cA</td>
								<td align="center">9.25 ± 1.50bA</td>
								<td align="center">10 ± 0.00bA</td>
								<td align="center">9.16 ± 1.67bA</td>
							</tr>
						</tbody>
					</table>
					<table-wrap-foot>
						<fn id="TFN1">
							<p>Note: means <italic>followed</italic> by the <italic>same letter do
									not differ</italic> statistically between themselves
									<italic>according to the Tukey test at</italic> 5 %.Lowercase
								letters are used to compare different concentrations within the same
								treated group and uppercase letters to compare the concentration of
								different treatments.</p>
						</fn>
						<fn id="TFN2">
							<p>Co<sup>-</sup>: negative control group, RME: raw meatpacking
								effluent, TME: treated meatpacking effluent, RTE: raw textile
								effluent, TTE: treated textile effluent</p>
						</fn>
					</table-wrap-foot>
				</table-wrap>
			</p>
			<p>
				<table-wrap id="t2">
					<label>TABLE II</label>
					<caption>
						<title>TOXICITY PERCENTAGE OF <italic>Artemia salina</italic> L. TREATED
							WITH DIFFERENT CONCENTRATIONS OF EFFLUENTS</title>
					</caption>
					<table frame="hsides" rules="groups">
						<colgroup>
							<col/>
							<col/>
							<col/>
							<col/>
							<col/>
						</colgroup>
						<tbody>
							<tr>
								<td align="center">Concentration</td>
								<td align="center">RME (%)</td>
								<td align="center">TME (%)</td>
								<td align="center">RTE (%)</td>
								<td align="center">TTE (%)</td>
							</tr>
							<tr>
								<td align="center">Co<sup>-</sup></td>
								<td align="center">2.5</td>
								<td align="center">2.5</td>
								<td align="center">2.5</td>
								<td align="center">2.5</td>
							</tr>
							<tr>
								<td align="center">3.1 %</td>
								<td align="center">20</td>
								<td align="center">10</td>
								<td align="center">5.0</td>
								<td align="center">19.57</td>
							</tr>
							<tr>
								<td align="center">6.2 %</td>
								<td align="center">7.5</td>
								<td align="center">10</td>
								<td align="center">7.5</td>
								<td align="center">34.17</td>
							</tr>
							<tr>
								<td align="center">12.5 %</td>
								<td align="center">9.15</td>
								<td align="center">17.5</td>
								<td align="center">12.5</td>
								<td align="center">0</td>
							</tr>
							<tr>
								<td align="center">25 %</td>
								<td align="center">7.5</td>
								<td align="center">5.0</td>
								<td align="center">7.5</td>
								<td align="center">3.55</td>
							</tr>
							<tr>
								<td align="center">50 %</td>
								<td align="center">32.07</td>
								<td align="center">5.0</td>
								<td align="center">22.5</td>
								<td align="center">24.75</td>
							</tr>
							<tr>
								<td align="center">100 %</td>
								<td align="center">76.6</td>
								<td align="center">92.5</td>
								<td align="center">100</td>
								<td align="center">91.65</td>
							</tr>
						</tbody>
					</table>
					<table-wrap-foot>
						<fn id="TFN3">
							<p>RME: raw meatpacking effluent, TME: treated meatpacking effluent,
								RTE: raw textile effluent, TTE: treated textile effluent,
									Co<sup>-</sup>: negative control group</p>
						</fn>
					</table-wrap-foot>
				</table-wrap>
			</p>
			<p>Thus, the highest concentrations of raw meatpacking effluents (50 % and 100 %) and
				treated meatpacking effluents (100 %) caused the mortality of organisms,
				statistically different from the negative control (<xref ref-type="table" rid="t1"
					>Table I</xref>), with percentage of toxicity form 32 to 92.5 %. In addition,
				the highest concentration of the raw and treated meatpacking efﬂuents was
				statistically different from the other concentrations evaluated in the same
				efﬂuents, indicating that the dilution is related to the non-toxicity of the
				efﬂuents to <italic>A. salina</italic>. A similar effect was observed by <xref
					ref-type="bibr" rid="B45">Silva (2011)</xref> in a toxicity test with
					<italic>Daphnia similis</italic>, where large concentrations of raw and treated
				meatpacking effluents were needed to trigger toxic effects. The work of Lied and
					<xref ref-type="bibr" rid="B23">Rodrigues (2012)</xref> with the effluents of a
				meatpacking plant and <italic>Daphnia magna</italic> as a bioindicator, also proved
				that the tested effluent samples showed toxicity. </p>
			<p>Despite the reduced toxic effects of the raw meatpacking efﬂuent in comparison to the
				treated efﬂuent, this is still toxic, so other treatments are evidently required in
				the plants to fully eliminate toxicity. Even though, the crude and treated effluents
				presented average number of similar dead organisms in the comparison of the same
				concentration for the two evaluated effluents. <xref ref-type="bibr" rid="B29"
					>Nogueira (2010)</xref> also demonstrated toxicity, corroborating these results;
				he used the acute toxicity test with <italic>Daphnia similis</italic> in the treated
				effluent of a meatpacking plant in different months of the year. </p>
			<p>For the textile effluent, the raw and treated effluents were statistically toxic in
				the highest concentration tested (100 %) (<xref ref-type="table" rid="t1">Table
					I</xref>), with a toxicity percentage &gt; 91 % (<xref ref-type="table" rid="t2"
					>Table II</xref>). Furthermore, there was no statistical difference in the
				number of dead organisms of the treated textile effluents when compared with the raw
				effluents in the different concentrations; however, the highest concentration of the
				raw and treated textile effluent was statistically different from the other
				concentrations evaluated in the same effluent, indicating that the dilution of the
				effluent allows the dilution of its possible toxic agents to <italic>A.
					salina</italic>. <xref ref-type="bibr" rid="B6">Bilal et al. (2016a)</xref> also
				confirmed the toxicity of a textile effluent using <italic>A. salina</italic> as
				bioindicator, but noted that after this effluent was treated with manganese peroxide
				in Chitosan spheres, the toxicity was reduced, unlike the findings of this study
				with the treatment applied by the plant with the coagulating agent aluminum sulfate.
					<xref ref-type="bibr" rid="B34">Punzi et al. (2015a)</xref> also showed that the
				use of biological treatment followed by photo-Fenton oxidation might have reduced
				the toxicity of treated textile effluents for the bioindicator <italic>A.
					salina</italic>, suggesting that advanced oxidation is effective in degrading
				organic compounds and removing the toxicity of textile effluents. In this way, data
				suggest that some additional treatment should be carried out in the textile effluent
				of the present study.</p>
			<p>In this study, the toxicity of effluents in high concentrations may be caused by the
				presence of detergents, which hinders the gas exchange between the water surface and
				the atmosphere, preventing oxygenation, possibly leading to the death of aquatic
				organisms and affecting and deconstructing the cell membranes of microorganisms
					(<xref ref-type="bibr" rid="B39">Romanelli 2004</xref>). A similar effect may be
				caused by a high content of oils and greases in the meatpacking effluent, which may
				cause toxicity to microorganisms and solidify at low temperatures, generating waste
				in the receiving body (<xref ref-type="bibr" rid="B38">Rigo 2004</xref>). According
				to <xref ref-type="bibr" rid="B20">Jadhav et al. (2010)</xref>, the major chemical
				pollutants present in textile wastewater are dyes containing carcinogenic amines,
				toxic heavy metals, pentachlorophenol, chlorine bleaching, halogen carriers, free
				formaldehyde, biocides, fire retardants, and softeners. These synthetic dyes do not
				degrade easily and have a long half-life, as can be observed by the high color
				values obtained in the physical and chemical analyses (<xref ref-type="table"
					rid="t3">Table III</xref>).</p>
			<p>
				<table-wrap id="t3">
					<label>TABLE III</label>
					<caption>
						<title>MEANS AND STANDARD DEVIATIONS OF THE PHYSICAL AND CHEMICAL ANALYSES
							OF THE EFFLUENTS</title>
					</caption>
					<table frame="hsides" rules="groups">
						<colgroup>
							<col/>
							<col/>
							<col/>
							<col/>
							<col/>
							<col/>
							<col/>
							<col/>
							<col/>
						</colgroup>
						<tbody>
							<tr>
								<td align="justify">Parameter</td>
								<td align="center">Unit</td>
								<td align="center">RME</td>
								<td align="center">TME</td>
								<td align="center">RTE</td>
								<td align="center">TTE</td>
								<td align="center">Regulation 430 (<xref ref-type="bibr" rid="B9"
										>CONAMA 2011</xref>)</td>
								<td align="center">Regulation 357 (<xref ref-type="bibr" rid="B8"
										>CONAMA 2005</xref>)</td>
								<td align="center">
									<xref ref-type="bibr" rid="B42">SEMA/IAP (2008)</xref>
								</td>
							</tr>
							<tr>
								<td align="justify">pH</td>
								<td align="center">-</td>
								<td align="center">8.55 ± 0.43a</td>
								<td align="center">7.92 ± 0.01a</td>
								<td align="center">9.29 ± 0.01a*</td>
								<td align="center">4.37 ± 0.07a*#</td>
								<td align="center">5.00 - 9.00</td>
								<td align="center">6.00 - 9.00</td>
								<td align="center">-</td>
							</tr>
							<tr>
								<td align="justify">COD</td>
								<td align="center">mg O<sub>2</sub>/L</td>
								<td align="center">206.33 ± 51.25a≠</td>
								<td align="center">121.33 ± 19.09a*</td>
								<td align="center">143.00 ± 0.00a</td>
								<td align="center">253.83 ± 122.00a*≠</td>
								<td align="center">Reduction 60 %</td>
								<td align="center">-</td>
								<td align="center">&lt; 200</td>
							</tr>
							<tr>
								<td align="justify">Total Solids</td>
								<td align="center">mg/L</td>
								<td align="center">1026.66 ± 110.15a*</td>
								<td align="center">296.66 ± 49.49b*</td>
								<td align="center">2260.00 ± 217.03a*</td>
								<td align="center">1433.33 ± 120.14b*</td>
								<td align="center">VA</td>
								<td align="center">-</td>
								<td align="center">-</td>
							</tr>
							<tr>
								<td align="justify">DO</td>
								<td align="center">mg O<sub>2</sub>/L</td>
								<td align="center">2.73 ± 0.00a#</td>
								<td align="center">7.30 ± 0.00a</td>
								<td align="center">6.19 ± 0.00a</td>
								<td align="center">5.87 ± 0.00a</td>
								<td align="center">-</td>
								<td align="center">≥ 5</td>
								<td align="center">-</td>
							</tr>
							<tr>
								<td align="justify">Temperature</td>
								<td align="center">ºC</td>
								<td align="center">28.40 ± 0.00a</td>
								<td align="center">28.4 ± 0.00a</td>
								<td align="center">26.90 ± 0.00a</td>
								<td align="center">26.60 ± 0.00a</td>
								<td align="center">&lt; 40</td>
								<td align="center">-</td>
								<td align="center">-</td>
							</tr>
							<tr>
								<td align="justify">Ammoniacal N</td>
								<td align="center">mg NH<sub>3</sub>/L</td>
								<td align="center">0.09 ± 0.01a</td>
								<td align="center">0.03 ± 0.00a</td>
								<td align="center">-</td>
								<td align="center">-</td>
								<td align="center">&lt; 20</td>
								<td align="center">≤ 2.00</td>
								<td align="center">-</td>
							</tr>
							<tr>
								<td align="justify">Chloride</td>
								<td align="center">mg Cl /L</td>
								<td align="center">-</td>
								<td align="center">-</td>
								<td align="center">1085.71 ± 73.71a#</td>
								<td align="center">882.14 ± 12.49a#</td>
								<td align="center">-</td>
								<td align="center">&lt; 250</td>
								<td align="center">-</td>
							</tr>
							<tr>
								<td align="justify">Alkalinity C</td>
								<td align="center">mg CaCO<sub>3</sub>/L</td>
								<td align="center">-</td>
								<td align="center">-</td>
								<td align="center">27.62 ± 1.41a</td>
								<td align="center">LDT</td>
								<td align="center">-</td>
								<td align="center">-</td>
								<td align="center">-</td>
							</tr>
							<tr>
								<td align="justify">Alkalinity B</td>
								<td align="center">mg CaCO<sub>3</sub>/L</td>
								<td align="center">-</td>
								<td align="center">-</td>
								<td align="center">330.34 ± 5.31a</td>
								<td align="center">164.78 ± 24.18a</td>
								<td align="center">-</td>
								<td align="center">-</td>
								<td align="center">-</td>
							</tr>
							<tr>
								<td align="justify">Alkalinity H</td>
								<td align="center">mg CaCO<sub>3</sub>/L</td>
								<td align="center">-</td>
								<td align="center">-</td>
								<td align="center">LDT</td>
								<td align="center">LDT</td>
								<td align="center">-</td>
								<td align="center">-</td>
								<td align="center">-</td>
							</tr>
							<tr>
								<td align="justify">Color</td>
								<td align="center">unPtCo</td>
								<td align="center">-</td>
								<td align="center">-</td>
								<td align="center">2960.14 ± 0.00a#</td>
								<td align="center">50.02 ± 0.00a</td>
								<td align="center">-</td>
								<td align="center">&lt; 75</td>
								<td align="center">-</td>
							</tr>
						</tbody>
					</table>
					<table-wrap-foot>
						<fn id="TFN4">
							<p>Means followed by the same letter in the rows do not differ
								statistically between themselves according to the Tukey test at 5
								%</p>
						</fn>
						<fn id="TFN5">
							<p>RME: raw meatpacking effluent, TME: treated meatpacking effluent,
								RTE: raw textile effluent, TTE: treated textile effluent COD:
								chemical oxygen demand, DO: dissolved oxygen, Ammoniacal N:
								ammoniacal nitrogen, Alkalinity C: carbonate alkalinity, Alkalinity
								B: bicarbonate alkalinity, Alkalinity H: hydroxide alkalinity. LDT:
								lower than the detection limit, VA: virtually absent</p>
						</fn>
						<fn id="TFN6">
							<p>*Data differ from the standards established by regulation 430 (<xref
									ref-type="bibr" rid="B9">CONAMA 2011</xref>), #data differs from
								the standards established by regulation 357 (<xref ref-type="bibr"
									rid="B8">CONAMA 2005</xref>) for class II fresh water, ≠does not
								comply with the legal requirements of <xref ref-type="bibr"
									rid="B42">SEMA/IAP (2008)</xref>
							</p>
						</fn>
					</table-wrap-foot>
				</table-wrap>
			</p>
			<p>According to <xref ref-type="bibr" rid="B47">Zahaira and Suteu (2012)</xref>,
				ecotoxicological studies indicated that over 18 % of 200 dyes tested in England
				showed significant inhibition of the respiration rate of wastewater bacteria from
				sewage. This can affect the entrance of light into the environment and the gas
				exchange, resulting in toxicity to different aquatic organisms, as already
				demonstrated in the works of <xref ref-type="bibr" rid="B35">Punzi et al.
					(2015b)</xref> with textile effluents in <italic>A. salina</italic> and
					<italic>Vibrio fischeriin.</italic><xref ref-type="bibr" rid="B31">Oliveira et
					al. (2018)</xref> also studied the ecotoxicity of textile dyes: both Direct
				Black 38 (azo dye) and Reactive Blue 15 (copper phthalocyanine dye) caused acute
				toxicity and genotoxicity to aquatic organisms; however, DB38 DB38 was the most
				toxic dye due to its lethal toxicity on <italic>A. salina</italic> and capacity to
				induce DNA damage in <italic>Daphnia magna</italic>.</p>
			<p>Moreover, meatpacking and textile effluents contain, among other compounds, organic
				matter, as demonstrated by the high values of chemical oxygen demand and total
				soluble solids of the effluents (<xref ref-type="table" rid="t3">Table III</xref>)
				evaluated in this study, which may affect the survival of organisms. With respect to
				the total solids content, despite the decline in content of the raw meatpacking and
				textile effluents compared to the treated effluents (RME = 1026.66 ± 110.15 mg/L;
				TME = 296.66 ± 49.49 mg/L; RTE = 2260.00 ± 217.03 mg/L; TTE = 1433.33 ± 120.14 mg/L)
					(<xref ref-type="table" rid="t3">Table III</xref>), values do not fall within
				those established by <xref ref-type="bibr" rid="B9">CONAMA (2011)</xref> in
				resolution 430 (virtually absent), which states the conditions and standards for the
				establishment of effluents. <xref ref-type="bibr" rid="B38">Rigo (2004)</xref>
				recorded higher total solids than the values found in this study for the meatpacking
				effluents after flotation (107 927.00 mg/L). This parameter, together with COD, is
				very important because solids may come from remnants of tissues and viscera or other
				industrial and treatment processes, which may directly affect the watersheds in
				which they are dumped. This may have resulted in the toxicity of the effluent under
				study at high concentrations for <italic>A. salina</italic>.</p>
			<p>COD is one of the most important physical and chemical parameters considered by the
				Secretariat of Environment and Water Resources of Paraná and the Environmental
				Institute of Paraná (SEMA/IAP), which state that the maximum COD for textile
				effluents is 200 mg/L (<xref ref-type="bibr" rid="B42">SEMA/IAP 2008</xref>). Thus,
				the treated textile effluents of this study fail to comply with this standard (TTE =
				253.83 ± 0.00 mg/L O<sub>2</sub>), and are very similar to those obtained by <xref
					ref-type="bibr" rid="B16">Hemachandra and Pathiratne (2016)</xref> for treated
				textile effluents of plant 1 (249.0 mg/L O<sub>2</sub>) and 2 (434.0 mg/L
					O<sub>2</sub>). Also, they fail to comply with the requirements of resolution
				430 (<xref ref-type="bibr" rid="B9">CONAMA 2011</xref>) to reduce 60 % of COD; in
				fact, the COD increased after the effluent was treated at the plant (RTE = 143.00 ±
				0.00 mg/L O<sub>2</sub>; TTE = 253.83 ± 0.00 mg/L O<sub>2</sub>), possibly due to
				the formation of byproducts that are more toxic than the initial composition of the
				effluents.</p>
			<p>In addition to the data presented above, the amount of DO did not increase in the
				treated textile effluents in comparison with the raw textile effluents (RTE = 6.19 ±
				0.00 mg/L O<sub>2</sub>; TTE = 5.87 ± 0.00 mg/L O<sub>2</sub>), although it was
				higher than the value established by <xref ref-type="bibr" rid="B8">CONAMA
					(2005)</xref> for water class II, which is the case of the river where this
				textile effluent was discharged (DO &gt; 5 mg/L O<sub>2</sub>). This finding is
				unfavorable, since DO is essential for the oxidation of organic matter and the
				respiration of the aerobic organisms that inhabit the location where the effluent is
				discharged (<xref ref-type="bibr" rid="B32">Pinto et al. 2010</xref>). In contrast,
				the DO of the present study was lower than the value observed by <xref
					ref-type="bibr" rid="B16">Hemachandra and Pathiratne (2016)</xref> for the
				treated effluents of plant 1 (7.8 mg/L O<sub>2</sub>) and greater than the value
				obtained for plant 2 (0.3 mg/L O<sub>2</sub>). The decrease in DO corroborates the
				increase in COD, since organisms use the DO to degrade organic matter. Evidently,
				the treatment used in the textile plant must be reviewed, since something is
				increasing the COD during the process, thus reducing the DO. According to <xref
					ref-type="bibr" rid="B4">Beltrame (2000)</xref>, residual dyes linked to organic
				and inorganic chemical assistants are responsible for the color, dissolved solids,
				and high COD values in textile effluents.</p>
			<p>Regarding the meatpacking effluents, data show that despite the 41.19 % reduction of
				COD when comparing the raw and treated meatpacking effluents (RME = 206.33 ± 51.25
				mg/L O<sub>2</sub>; TME = 121.33 ± 19.09 mg/L O<sub>2</sub>) (<xref ref-type="table"
					rid="t3">Table III</xref>), the plant also failed to reach the established legal
				values (<xref ref-type="bibr" rid="B9">CONAMA 2011</xref>), which stipulate that
				this parameter should be reduced by 60 % (from raw to treated effluents). However,
				the treated effluent reached the limit established by <xref ref-type="bibr"
					rid="B42">SEMA/IAP (2008)</xref> (COD &lt; 200 mg/L O<sub>2</sub>) for discharge
				into water bodies. <xref ref-type="bibr" rid="B36">Radoll (2013)</xref> obtained
				more satisfactory results, with a more than 60 % reduction in the treated effluents
				of the studied meatpacking plants and lower COD values (plant A = 24 mg/L
					O<sub>2</sub>, plant B = 37 mg/L O<sub>2</sub>). These data may be justified by
				the greater number of treatment steps of plants A and B compared to the present
				study. In contrast, the COD values of the raw effluents in the study of <xref
					ref-type="bibr" rid="B36">Radoll (2013)</xref> (plant A = 8680 mg/L
					O<sub>2</sub>, plant B = 7620 mg/L O<sub>2</sub>) were higher than those of the
				present study. According to <xref ref-type="bibr" rid="B29">Nogueira (2010)</xref>,
				COD can be affected by blood, fat, feathers (in the case of poultry
				slaughterhouses), scraps of fabric, viscera, and gizzards, all of which directly
				affect regional ecosystems if discarded into rivers, and have resulted in the
				toxicity for <italic>A. salina</italic> of the raw and treated textile effluents,
				including 50 % concentrations of the raw effluent.</p>
			<p>However, in relation to the OD content, after the effluents were treated in the
				meatpacking plants where the samples of this study were collected, these effluents
				had higher DO levels (RME = 2.73 ± 0.00 mg/L O<sub>2</sub>; TME = 7.30 ± 0.00 mg/L
					O<sub>2</sub>), and a value within the minimum value established by <xref
					ref-type="bibr" rid="B8">CONAMA (2005)</xref> for class 2 fresh water (DO &gt;
				5.0 mg/L O<sub>2</sub>), the same class as the receiving body of the plant’s
				effluents. <xref ref-type="bibr" rid="B36">Radoll (2013)</xref> also noted that
				after the meatpacking effluent was treated, there was an increase in DO (plant A,
				raw = 0.34 mg/L O<sub>2</sub>, treated = 1.27 mg/L O<sub>2</sub>; plant B, raw =
				0.28 mg/L O<sub>2</sub>, treated = 1.44 mg/L O<sub>2</sub>), although the increase
				in DO of the effluents in this study was greater. This result can be justified by
				the lower COD values of this study, as a lower amount of oxygen is required for its
				degradation; however, this can also be related to the adopted treatment process.</p>
			<p>In general, data of the physicochemical analysis (<xref ref-type="table" rid="t3"
					>Table III</xref>) show the efficiency of the treatment applied by the
				meatpacking plant. According to these data, there was a reduction of pH, ammoniacal
				nitrogen, COD, and total solids, as previously discussed. However, only the data of
				pH (RME = 8.55 ± 0.43; TME = 7.92 ± 0.01), temperature (RME = 28.4 ± 0 ºC; TME =
				28.4 ± 0 ºC), and ammoniacal nitrogen (RME = 0.09 ± 0.01 mg NH<sub>3</sub>/L; TME =
				0.00 ± 0.03, mg NH<sub>3</sub>/L) are within the levels established by resolution
				430 (<xref ref-type="bibr" rid="B9">CONAMA, 2011</xref>) (pH between 5.0-9.0,
				temperature under 40 ºC, ammoniacal nitrogen under 20 mg NH<sub>3</sub>/L). In
				accordance with this resolution, the above mentioned parameters will not affect the
				water quality of rivers that receive the treated effluents since they are below
				those established by resolution 357 (<xref ref-type="bibr" rid="B8">CONAMA
					2005</xref>) for class II fresh water (pH = 6-9, ammoniacal nitrogen below 2.0
				mg NH<sub>3</sub>/L).</p>
			<p>The pH of the meatpacking effluent in this study is higher than the pH found by <xref
					ref-type="bibr" rid="B36">Radoll (2013)</xref> in the assessment of two
				meatpacking plants. The author recorded a pH of 7.02 (plant A) and 6.80 (plant B) in
				the raw effluents, as well as a pH of 6.77 in the effluent of plant A with a
				treatment consisting of an anaerobic lagoon, two facultative ponds in series and a
				pond of maturation, and of 6.30 in the effluent of plant B, whose treatment
				consisted of an anaerobic lagoon followed by a facultative lagoon and two lagoons
				with aeration. In contrast, the temperatures of the raw effluents of the plants
				studied by <xref ref-type="bibr" rid="B36">Radoll (2013)</xref> (plant A = 27.5 ºC,
				plant B = 26.7 ºC) were similar to those of the present study. With respect to total
				nitrogen, <xref ref-type="bibr" rid="B37">Ribeiro et al. (2013)</xref> found higher
				values for the treated effluent of a bovine slaughterhouse (1.53 mg
				NH<sub>3</sub>/L) than the values recorded in the present study.</p>
			<p>The physical and chemical analyses of the textile effluents (<xref ref-type="table"
					rid="t3">Table III</xref>) also show that after the treatment of raw effluents,
				there was a reduction of pH (RTE = 9.28 ± 0.01; TTE = 4.37 ± 0.07), concentration of
				chlorides (RTE = 1085.71 ± 73.71 mg/L Cl; TTE = 882.14 ± 12.49 mg/L Cl), carbonate
				alkalinity (RTE = 27.62 ± 1.41 mg/L CaCO<sub>3</sub>; TTE = 0.00 ± 0.00 mg/L
					CaCO<sub>3</sub>), bicarbonate alkalinity (RTE = 330.34 ± 5.31 mg/L
					CaCO<sub>3</sub>; TTE = 164.78 ± 24.18 mg/L CaCO<sub>3</sub>), color (RTE =
				1960.14 ± 0.00 unPtCo; TTE = 50.02 ± 0.00 unPtCo), and total solids, the latter
				showing a statistical difference using the Tukey test. However, only the data of
				temperature (RTE = 26.90 ± 0.00 ºC; TTE = 26.60 ± 0.00 ºC) and color were within the
				levels established by resolution 430 (<xref ref-type="bibr" rid="B9">CONAMA
					2011</xref>) (temperature under 40 ºC, color under 75 mg unPtCo). Furthermore,
				the data of the present study are similar to those obtained by <xref ref-type="bibr"
					rid="B16">Hemachandra and Pathiratne (2016)</xref> for treated textile effluents
				(plant 1 = 30.5 ºC, plant 2 = 29.3 ºC). </p>
			<p>It is worth mentioning that the reduction in pH of the raw efﬂuent is outside the
				limits established by resolution 430 (pH between 5.0-9.0) (<xref ref-type="bibr"
					rid="B9">CONAMA 2011</xref>). Possibly, some stage of the treatment applied by
				the textile industry resulted in the decrease of the pH. These data may have
				contributed to the toxicity of the treated effluent since low pH (acids) hinder the
				survival of organisms (<xref ref-type="bibr" rid="B14">Fuhrmann et al. 2019</xref>).
				Besides that, in comparison to other textile effluents, such as those syudied by
					<xref ref-type="bibr" rid="B16">Hemachandra and Pathiratne (2016)</xref> for two
				different plants (plant 1 = 8.38, plant 2 = 8.15), the pH of the treated textile
				effluents in this study had the lowest value of all tested effluents.</p>
			<p>Thus, the discharge of these effluents in rivers can cause changes in the ecological
				health of the receiving waters, hence the importance of aggregating ecotoxicological
				analyses to the verification of physical and chemical parameters (<xref
					ref-type="bibr" rid="B16">Hemachandra and Pathiratne 2016</xref>), mainly
				because effluents may still contain substances potentially toxic to aquatic
				organisms, even though they remain within the limits of legislation in physical and
				chemical terms, as demonstrated in the immobility/mortality test with <italic>A.
					salina</italic>.</p>
			<p>However, organisms of different trophic levels can respond in various ways to toxic
				pollutants, so the use of different bioindicators in the ecotoxicological
				evaluations makes it possible the know the level at which the pollutant interacts
				with the organism and the susceptibility of the latter (<xref ref-type="bibr"
					rid="B3">Arias et al. 2007</xref>, <xref ref-type="bibr" rid="B22">Libralato et
					al. 2016</xref>).</p>
			<p>Thus, the results of the cytotoxicity test with <italic>A. cepa</italic> (<xref
					ref-type="fig" rid="f1">Fig. 1</xref>) showed that neither the raw or treated
				effluents (24 h) of the meatpacking or textile plants had cytotoxic effects on this
				vegetal bioindicator, since it presented mitotic indices similar to those of the
				negative control (24 h) and the control of the bulb itself (0 h), and did not
				influence the cell divisions of the onion. In addition, the observation of the
				different mitotic phases (<xref ref-type="table" rid="t4">Table IV</xref>) and the
				percentage of phase index found in the raw and treated effluents of the meatpacking
				and textile plants, show that even within the different phases of the mitotic cell
				cycle (interphase, prophase, metaphase, anaphase, and telophase), there was no
				statistical difference between the control group (24 h), the control of the bulb (0
				h) and the groups treated (24 h) when compared with the Tukey test (α = 0.05). In
				addition, no statistical change was observed between the mitotic indices (<xref
					ref-type="fig" rid="f1">Fig. 1</xref>) and the phase index (<xref
					ref-type="table" rid="t5">Table V</xref>) between raw and treated effluents or
				between the textile and the meatpacking effluents, despite differences in the
				physical and chemical analyses (<xref ref-type="table" rid="t3">Table III</xref>) of
				these studied effluents.</p>
			<p>
				<fig id="f1">
					<label>Fig. 1</label>
					<caption>
						<title>Mitotic index percentages (MI %) for the collection times of 0, 24,
							and 48 h. Co<sup>-</sup>: negative control group, RME: raw meatpacking
							effluents, TME: treated meatpacking effluents, RTE: raw textile
							effluents, TTE: treated textile effluents. Means followed by the same
							letter in the columns do not differ statistically between themselves
							according to the Tukey test at 5 %</title>
					</caption>
					<graphic xlink:href="0188-4999-rica-36-02-399-gf1.png"/>
				</fig>
			</p>
			<p>
				<table-wrap id="t4">
					<label>TABLE IV</label>
					<caption>
						<title>DIFFERENT MITOTIC PHASES FOR THE COLLECTION TIMES OF 0, 24, AND 48
							h</title>
					</caption>
					<table frame="hsides" rules="groups">
						<colgroup>
							<col/>
							<col span="3"/>
							<col span="3"/>
							<col span="3"/>
							<col span="3"/>
							<col span="3"/>
						</colgroup>
						<tbody>
							<tr>
								<td align="justify" rowspan="2">Mitotic phases</td>
								<td align="center" colspan="3">Co<sup>-</sup></td>
								<td align="center" colspan="3">RME </td>
								<td align="center" colspan="3">TME </td>
								<td align="center" colspan="3">RTE </td>
								<td align="center" colspan="3">TTE </td>
							</tr>
							<tr>
								<td align="center">0 h</td>
								<td align="center">24 h</td>
								<td align="center">48 h</td>
								<td align="center">0 h</td>
								<td align="center">24 h</td>
								<td align="center">48 h</td>
								<td align="center">0 h</td>
								<td align="center">24 h</td>
								<td align="center">48 h</td>
								<td align="center">0 h</td>
								<td align="center">24 h</td>
								<td align="center">48 h</td>
								<td align="center">0 h</td>
								<td align="center">24 h</td>
								<td align="center">48 h</td>
							</tr>
							<tr>
								<td align="justify">Interphase</td>
								<td align="center">4980</td>
								<td align="center">4985</td>
								<td align="center">4946</td>
								<td align="center">4979</td>
								<td align="center">4991</td>
								<td align="center">4989</td>
								<td align="center">4944</td>
								<td align="center">4962</td>
								<td align="center">4928</td>
								<td align="center">4947</td>
								<td align="center">4980</td>
								<td align="center">4982</td>
								<td align="center">4931</td>
								<td align="center">4950</td>
								<td align="center">4935</td>
							</tr>
							<tr>
								<td align="justify">Prophase</td>
								<td align="center">04</td>
								<td align="center">06</td>
								<td align="center">14</td>
								<td align="center">05</td>
								<td align="center">04</td>
								<td align="center">0</td>
								<td align="center">23</td>
								<td align="center">12</td>
								<td align="center">30</td>
								<td align="center">14</td>
								<td align="center">06</td>
								<td align="center">07</td>
								<td align="center">24</td>
								<td align="center">23</td>
								<td align="center">16</td>
							</tr>
							<tr>
								<td align="justify">Metaphase</td>
								<td align="center">07</td>
								<td align="center">05</td>
								<td align="center">15</td>
								<td align="center">05</td>
								<td align="center">02</td>
								<td align="center">01</td>
								<td align="center">16</td>
								<td align="center">11</td>
								<td align="center">33</td>
								<td align="center">11</td>
								<td align="center">02</td>
								<td align="center">01</td>
								<td align="center">21</td>
								<td align="center">15</td>
								<td align="center">33</td>
							</tr>
							<tr>
								<td align="justify">Anaphase</td>
								<td align="center">04</td>
								<td align="center">03</td>
								<td align="center">20</td>
								<td align="center">08</td>
								<td align="center">01</td>
								<td align="center">05</td>
								<td align="center">13</td>
								<td align="center">09</td>
								<td align="center">07</td>
								<td align="center">27</td>
								<td align="center">02</td>
								<td align="center">06</td>
								<td align="center">16</td>
								<td align="center">11</td>
								<td align="center">11</td>
							</tr>
							<tr>
								<td align="justify">Telophase</td>
								<td align="center">05</td>
								<td align="center">01</td>
								<td align="center">05</td>
								<td align="center">03</td>
								<td align="center">02</td>
								<td align="center">05</td>
								<td align="center">04</td>
								<td align="center">06</td>
								<td align="center">02</td>
								<td align="center">01</td>
								<td align="center">10</td>
								<td align="center">04</td>
								<td align="center">08</td>
								<td align="center">02</td>
								<td align="center">05</td>
							</tr>
						</tbody>
					</table>
					<table-wrap-foot>
						<fn id="TFN7">
							<p>Co<sup>-</sup>: negative control group, RME: raw meatpacking
								effluents, TME: treated meatpacking effluents, RTE: raw textile
								effluents, TTE: treated textile effluents</p>
						</fn>
					</table-wrap-foot>
				</table-wrap>
			</p>
			<p>
				<table-wrap id="t5">
					<label>TABLE V</label>
					<caption>
						<title>PHASE INDEX (%)FOR THE COLLECTION TIMES OF 0, 24, AND 48 h</title>
					</caption>
					<table frame="hsides" rules="groups">
						<colgroup>
							<col/>
							<col span="3"/>
							<col span="3"/>
							<col span="3"/>
							<col span="3"/>
							<col span="3"/>
						</colgroup>
						<tbody>
							<tr>
								<td align="justify" rowspan="2">Mitotic phases</td>
								<td align="center" colspan="3">Co<sup>-</sup></td>
								<td align="center" colspan="3">RME </td>
								<td align="center" colspan="3">TME </td>
								<td align="center" colspan="3">RTE </td>
								<td align="center" colspan="3">TTE </td>
							</tr>
							<tr>
								<td align="center">0 h</td>
								<td align="center">24 h</td>
								<td align="center">48 h</td>
								<td align="center">0 h</td>
								<td align="center">24 h</td>
								<td align="center">48 h</td>
								<td align="center">0 h</td>
								<td align="center">24 h</td>
								<td align="center">48 h</td>
								<td align="center">0 h</td>
								<td align="center">24 h</td>
								<td align="center">48 h</td>
								<td align="center">0 h</td>
								<td align="center">24 h</td>
								<td align="justify">48 h</td>
							</tr>
							<tr>
								<td align="justify">Interphase</td>
								<td align="center">99.60a</td>
								<td align="center">99.70a</td>
								<td align="center">98.92a</td>
								<td align="center">99.58a</td>
								<td align="center">99.82a</td>
								<td align="center">99.78a</td>
								<td align="center">98.88a</td>
								<td align="center">99.24a</td>
								<td align="center">98.56a</td>
								<td align="center">98.94a</td>
								<td align="center">99.6a</td>
								<td align="center">99.64a</td>
								<td align="center">98.62a</td>
								<td align="center">99.00a</td>
								<td align="justify">98.70a</td>
							</tr>
							<tr>
								<td align="justify">Prophase</td>
								<td align="center">0.08bc</td>
								<td align="center">0.12bc</td>
								<td align="center">0.28abc</td>
								<td align="center">0.10bc</td>
								<td align="center">0.08bc</td>
								<td align="center">0c</td>
								<td align="center">0.46abc</td>
								<td align="center">0.24abc</td>
								<td align="center">0.60a</td>
								<td align="center">0.28ab</td>
								<td align="center">0.12abc</td>
								<td align="center">0.14abc</td>
								<td align="center">0.48abc</td>
								<td align="center">0.46bc</td>
								<td align="justify">0.32abc</td>
							</tr>
							<tr>
								<td align="justify">Metaphase</td>
								<td align="center">0.14a</td>
								<td align="center">0.10a</td>
								<td align="center">0.30a</td>
								<td align="center">0.10a</td>
								<td align="center">0.04a</td>
								<td align="center">0.02a</td>
								<td align="center">0.32a</td>
								<td align="center">0.22a</td>
								<td align="center">0.66a</td>
								<td align="center">0.22a</td>
								<td align="center">0.04a</td>
								<td align="center">0.02a</td>
								<td align="center">0.42a</td>
								<td align="center">0.30a</td>
								<td align="justify">0.66a</td>
							</tr>
							<tr>
								<td align="justify">Anaphase</td>
								<td align="center">0.08a</td>
								<td align="center">0.06a</td>
								<td align="center">0.40a</td>
								<td align="center">0.16a</td>
								<td align="center">0.02a</td>
								<td align="center">0.10a</td>
								<td align="center">0.26a</td>
								<td align="center">0.18a</td>
								<td align="center">0.14a</td>
								<td align="center">0.54a</td>
								<td align="center">0.04a</td>
								<td align="center">0.12a</td>
								<td align="center">0.32a</td>
								<td align="center">0.22a</td>
								<td align="justify">0.22a</td>
							</tr>
							<tr>
								<td align="justify">Telophase</td>
								<td align="center">0.10a</td>
								<td align="center">0.02a</td>
								<td align="center">0.10a</td>
								<td align="center">0.06a</td>
								<td align="center">0.04a</td>
								<td align="center">0.10a</td>
								<td align="center">0.08a</td>
								<td align="center">0.12a</td>
								<td align="center">0.04a</td>
								<td align="center">0.02a</td>
								<td align="center">0.20a</td>
								<td align="center">0.08a</td>
								<td align="center">0.16a</td>
								<td align="center">0.04a</td>
								<td align="justify">0.10a</td>
							</tr>
						</tbody>
					</table>
					<table-wrap-foot>
						<fn id="TFN8">
							<p>Note: Means <italic>followed</italic> by the <italic>same letter in
									rows do not differ</italic> statistically between themselves
									<italic>according to the Tukey test at</italic> 5 %, for each
								phase of mitotic cell division</p>
						</fn>
						<fn id="TFN9">
							<p>Co<sup>-</sup>: negative control group, RME: raw meatpacking
								effluents, TME: treated meatpacking effluents, RTE: raw textile
								effluents, TTE: treated textile effluents</p>
						</fn>
					</table-wrap-foot>
				</table-wrap>
			</p>
			<p>
				<xref ref-type="bibr" rid="B36">Radoll (2013)</xref> obtained different results than
				those of this study when assessing the raw and treated effluents of two meatpacking
				plants and observed significant genotoxic damage of these effluents on fish used as
				bioindicators. In the case of textile effluents, <xref ref-type="bibr" rid="B16"
					>Hemachandra and Pathiratne (2016)</xref> also obtained results different to
				those in study, since their tests showed cytotoxicity even at dilutions of 1:8 of
				treated textile effluents with <italic>A. cepa</italic> and <italic>Oreochromis
					niloticus</italic> As bioindicators.</p>
			<p>However, it can be observed that the mitotic index percentage (that is, of the onion
				cell divisions) dropped from the raw textile effluent (MI = 1.01 %) to the treated
				effluent (MI = 0.40 %). Moreover, in the recovery time of this bioindicator, it was
				the only treatment that did not show a reduction of the mitotic index from 24 h (MI
				= 0.40 %) to 48 h (MI = 0.36 %). These results may be justified by the decrease in
				the pH from the raw to the treated textile effluent, as well as the increase in COD
					(<xref ref-type="table" rid="t3">Table III</xref>), which may have enlarged the
				amount of potentially toxic substances in the effluent, indicating that new
				treatment steps should be applied or even improved in order to avoid toxicity
				thereof. <xref ref-type="bibr" rid="B1">Alvim et al. (2011)</xref> used <italic>A.
					cepa</italic> to assess the cytotoxic potential of raw and treated effluents of
				two textile plants in southern Minas Gerais and noted a lower mitotic index in all
				the treated effluent concentrations when compared to raw effluent concentrations. In
				contrast, Bilal et al. (<xref ref-type="bibr" rid="B6">2016a</xref>, <xref
					ref-type="bibr" rid="B7">b</xref>) obtained different results to those of this
				study when treating textile effluents with manganese peroxide encapsulated in
				Ca-alginate or in chitosan beads. In this case, the treated effluent increased the
				average mitotic index percentage of the onion roots. <xref ref-type="bibr" rid="B18"
					>Iqbal and Nisar (2015)</xref> treated textile effluents with gamma radiation
				and hydrogen peroxide, finding a 50 % reduction in toxicity from the raw effluent to
				the treated effluent. These studies indicate that adequate treatment of effluents
				can reduce the formation of cytotoxic agents or eliminate existing ones.</p>
			<p>Therefore, considering the importance that physical, chemical and toxicological
				parameters remain within values that are not harmful to the environment, it is
				indicated that additional treatments should be carried out in textile and
				meatpacking effluents. The analyses carried out in this study are fundamental to
				develop and improve treatments and the results contribute to highlight the
				importance for industries in regard to effluent disposals and the observance of
				standards. </p>
		</sec>
		<sec sec-type="conclusions">
			<title>CONCLUSION</title>
			<p>Results show that only the lowest concentrations (3.1, 6.2, 12.5 and 25 %) of the raw
				and treated effluents of the meatpacking and textile industries were not toxic to
					<italic>A. salina</italic>. The highest concentrations of raw meatpacking
				effluents (50 and 100 %), the highest concentration of raw textile effluents (100 %)
				and the highest concentration of the treated meatpacking and textile effluents (100
				%) were toxic to this bioindicator. Thus, it is evident that additional treatment
				measures are required in both plants to fully eliminate toxicity, mainly because the
				values of physical and chemical parameters (total solids content [for treated
				meatpacking and textile effluents], COD [for treated meatpacking and textile
				effluents], chlorides, carbonate alkalinity and bicarbonate alkalinity [for treated
				textile effluents]) are not being effectively reduced in order to meet the levels
				required by the current legislation. Other parameters (pH, temperature, ammoniacal
				nitrogen and color) remain within the limits established by the national
				legislation. </p>
			<p>None of the treatments had a cytotoxic effect on <italic>A. cepa</italic>, because
				they did not change statistically the mitotic index and the index of phases.
				However, the mitotic index percentage dropped from the raw textile effluent to the
				treated effluent. Thus, the results stress the importance of effectively treating
				effluents to prevent environmental harm and health problems in the area of
				discharge.</p>
			<p>This study suggests that some additional treatments should be carried out in the
				textile and meatpacking effluents. Monitoring the effectiveness of these measures is
				fundamental for the development of new techniques that enhance the results of
				industrial effluents treatment. This may reflect in the development of new
				treatments that are more effective and have a better cost-benefit.</p>
		</sec>
	</body>
	<back>
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