*A. cepa* has been indicated as an efficient bioindicator in genotoxicity testing, due to cellular proliferation kinetics, the rapid growth of its roots, large numbers of cells in division, high tolerance to different cultivation conditions, year-round availability, easy management and reduced number of large chromosomes (2 n = 16) [20-21]. The *Allium cepa* test has been used since the 1930s [22] and subsequently standardized by [20] in other studies (1985). Since then, many studies adopt this methodology and the results confirm its efficiency in detecting the effects caused by toxic substances found in the environment. A literature review by [23] shows that 148 chemicals had the clastogenic potential when tested through the *Allium* test. Among Among the several classes of contaminants detected by the sensitivity of the *Allium* test are: heavy metals, domestic and industrial sewage, landfill extracts and samples of water from rivers and lakes, whose solutions encompass a complex mixture of substances of different compositions, thus attesting the sensitivity and efficacy of this bioassay. Some heavy metals, such as iron, nickel and chrome, are among the contaminants commonly found in the waters of rivers and lakes, act as toxic agents, and can influence the mitotic rates, induce chromosomal aberrations and the formation of micronuclei in tests conducted with *Allium cepa*. Certain metals in aqueous solution can cross the cellular membrane or enter through phagocytosis or pinocytosis and can cause damages to the DNA molecule structure [25].

The first studies conducted in *Allium cepa* roots were performed by [23] when investigating the toxic action of colchicine. Since then, numerous works have used this protocol to estimate the environmental danger in several situations.

[26] when assessing samples of domestic sewage from the residual water from a municipal water treatment station, observed a dramatic reduction in the mitotic index, so that it was impossible to count chromosome aberration; but the dilution is a factor that can influence these parameters. Similar results were also observed by [27] and [28] when estimating the genotoxicity in pharmaceutical/hospital effluents samples and [29] when assessing water samples from a lake environmentally affected with heavy metals. Other works also observed the environmental impact of the presence of heavy metals in rivers, lakes and soil aqueous extracts [30-34]. In addition to the domestic sewage [26,35-37], industrial effluents are also historically impacting: [38] submitted seeds of *A. cepa* to the effluent from a textile industry and observed a mutagenic effect, the type of cell damage may be transmitted to subsequent generations, possibly affecting the organism as a whole, as well as the local biota exposed to the effluent discharge. If the damage results in cell death, the development of the organism may be affected, which could also lead to its death. Chromosome aberration assay was carried out in *A. cepa* meristematic cells exposed to the Guaecá river waters, located in the city of São Sebastião, SP, Brazil, which had its waters impacted by an oil pipeline leak. Analyses of the aberration types showed clastogenic and aneugenic effects for the roots exposed to the polluted waters from Guaecá river, besides the induction of cell death. Probably all the observed effects were induced by the petroleum hydrocarbons derived from the oil leakage [39]. In the work of [40], the general toxicity (root growth inhibition and malformation) and genotoxicity (induction of chromosome aberrations in root cells) of an oil field wastewater in the Nigeria have been investigated by the *Allium* test. The wastewater is mitodepressive and increased significantly the frequency of chromosome aberrations in root cells (sticky chromosomes, c-mitosis, spindle multipolarity, bridges and fragments). At lower concentrations c-mitosis was the most common aberration. The suitability of the *Allium* test in genotoxicity screening is highlighted and the impact and significance of positive results on the environment and human health should be discussed.

Bioassays with Plants in the Monitoring of Water Quality 325

spindle poisoning which induces an anomalous disjunction of chromosome at anaphase, has usually been considered a genotoxic indicator. On the other hand, chemicals substances can induce micronuclei promoting changes in the achromatic fuse (aneugenic effect). Micronuclei arise when the lost genetic material is involved by a nuclear envelope, independent of nuclear envelope of the principal nucleus [48]. When the interphase cell is exposed to environmental pollutants, damage is produced either in whole chromosomes - in G1, or damage in individual chromatids - S and G2 phases. Anomalies typically found in S phase are gaps and chromatid breaks [49]. Moreover, during mitosis, all types of AC may be found. Chromosome breaks, bridges and lagging chromosomes may result from sticks and are considered to induce aneuploid and polyploid cells. The stick form can be related to the depolymerization of DNA,

the dissolution of nuclear proteins or with an increase in chromosome condensation.

**Figure 2.** Mitotic anomalies and micronucleus observed in root meristematic cells of *Allium cepa* exposed to water samples of the Paraiba do Sul river - Brazil: a. chromosome bridges; b. not identified;

The use of *Allium cepa* as a test system was introduced by [23], when the effects of colchicine were investigated. This author defined it as an inactivation of segments achromatic spindle fibers by condensed chromosomes randomly dispersed throughout the cytoplasm cell. The formation of c-mitotic inhibitors relates to fibers of the spindle. Calmodulin, involved in regulating the concentration of calcium in the cell was specifically localized in the zone achromatic of mitotic cells, suggesting its involvement in the movement of chromosomes by controlling the polymerization and depolymerization of microtubules [50]. The storage irregular calcium in the cell inhibits the polymerization of microtubules, causing the formation of c-mitosis [51]. The c-mitosis may be defined, as well as the permanence of the equatorial plate anaphase chromosomes rather than move to their respective poles [21]. Cmitosis results from inactivation of the mitotic spindle followed by a random scattering of

The chromosome stickiness arises from improper folding of the chromosome fiber into single chromatids and chromosomes. As a result there is an intermingling of the fibers, and the chromosomes become attached to each other by means of subchromatid bridges. Chromosome stickiness has generally been inflicted by highly toxic agents, usually of an irreversible type, and probably leads cells to death [18,46]. Chromosome breaks - Any descriptive classification must include a category for the break or discontinuity, a simple severance of the chromosome or chromatid to give an acentric fragment, and which is not clearly associated with any exchange process [52]. Usually, the mitotic anomalie is the most

The chromosome bridges can involve one or more chromosomes. Bridges associated with

c. multipolar anaphase; d. c-mitose; e. stickiness; f. micronuclei [47].

the condensed chromosomes [46].

frequently observed in studies conducted with *A. cepa*.

stickiness promote a usually irreversible toxic effects.

The uses of medicinal plants have always been part of human culture. The World Health Organization estimates that up to 80% of the world's population relies on traditional medicinal system for some aspect of primary health care. However, there are few reports on the toxicological properties of most medicinal plants especially, their mutagenicity and carcinogenicity. In this study, nine medicinal plants had their mutagenic potential assessed; the extracts inhibited the root growth, and promoted a mitodepressive effect and induction of chromosomal aberrations in *A. cepa* [41]. In other studies, plant extracts are assessed for the toxicity of their potential medicines: [42] detected a cytotoxic and genotoxic action when assessing extracts of *Inula viscosa*. [43] assessed the toxic effects of five medicinal plants, all the tested extracts were observed to have mitodepressive effects on cell division and induced mitotic spindle disturbance in *Allium cepa*.

### **3.2. Evaluated parameters in** *Allium cepa* **test**

Among the plants commonly used as indicators for studies of potential toxicity of river water, *Allium cepa* L. constitutes a convenient system for the analysis of anatomical (root growth, deformity, twist) and microscopic parameters (chromosome abnormalities, altered mitotic index (MI), and micronucleus (MN) formation) [44]. Root growth and mitotic index are parameters of the cytotoxicity; micronucleus and chromosome abnormalities parameters of genotoxicity. The mitotic index is a parameter that allows to estimate the frequency of cells in cell division, reflects cell proliferation and is regarded as an important parameter when determining the rate of plant root growth. Therefore, a low mitotic index (mitodepressive effect) characterize a few dividing cells [45-46] and is more frequently reported in the literature. This parameter predicts the root growth behavior, which is conducted by the frequency of cellular division in the root tissue. This index has been clearly correlated to the root length in the *A. cepa* test, as for instance decreasing in value with increasing concentrations of toxic metals like chromium and cadmium [46].

Chromosomal aberration types detected may depend on DNA lesions and the cell cycle phase that is considered [46]. The mitotic anomalies often detected by the *Allium* test are: chromosome stickiness, multipolar anaphase, anaphasic bridge, c–mitosis or c–metaphase, and micronuclei (Figure 2).

The induction of micronuclei in root meristems of *A. cepa* or in any cell of any other organism is the manifestation of chromosome damage and disturbance of the mitotic process. The micronucleus is formed by the development of a new membrane around the chromatin matter that failed to move to either pole during the anaphase of mitosis. Such chromatin matter arises either from anomalous disjunction of chromosomes due to spindle abnormalities or the breakage of chromosomes resulting in formation of acentric fragments, dicentric chromosomes and chromatin bridges. Thus, the induction of micronuclei may suggest that the environmental pollutant is either a spindle inhibitor or a clastogen. Induction of micronucleus formation, the outcome of chromosome breaks/fragments or spindle poisoning which induces an anomalous disjunction of chromosome at anaphase, has usually been considered a genotoxic indicator. On the other hand, chemicals substances can induce micronuclei promoting changes in the achromatic fuse (aneugenic effect). Micronuclei arise when the lost genetic material is involved by a nuclear envelope, independent of nuclear envelope of the principal nucleus [48]. When the interphase cell is exposed to environmental pollutants, damage is produced either in whole chromosomes - in G1, or damage in individual chromatids - S and G2 phases. Anomalies typically found in S phase are gaps and chromatid breaks [49]. Moreover, during mitosis, all types of AC may be found. Chromosome breaks, bridges and lagging chromosomes may result from sticks and are considered to induce aneuploid and polyploid cells. The stick form can be related to the depolymerization of DNA, the dissolution of nuclear proteins or with an increase in chromosome condensation.

324 Water Treatment

lower concentrations c-mitosis was the most common aberration. The suitability of the *Allium* test in genotoxicity screening is highlighted and the impact and significance of

The uses of medicinal plants have always been part of human culture. The World Health Organization estimates that up to 80% of the world's population relies on traditional medicinal system for some aspect of primary health care. However, there are few reports on the toxicological properties of most medicinal plants especially, their mutagenicity and carcinogenicity. In this study, nine medicinal plants had their mutagenic potential assessed; the extracts inhibited the root growth, and promoted a mitodepressive effect and induction of chromosomal aberrations in *A. cepa* [41]. In other studies, plant extracts are assessed for the toxicity of their potential medicines: [42] detected a cytotoxic and genotoxic action when assessing extracts of *Inula viscosa*. [43] assessed the toxic effects of five medicinal plants, all the tested extracts were observed to have mitodepressive effects on cell division and

Among the plants commonly used as indicators for studies of potential toxicity of river water, *Allium cepa* L. constitutes a convenient system for the analysis of anatomical (root growth, deformity, twist) and microscopic parameters (chromosome abnormalities, altered mitotic index (MI), and micronucleus (MN) formation) [44]. Root growth and mitotic index are parameters of the cytotoxicity; micronucleus and chromosome abnormalities parameters of genotoxicity. The mitotic index is a parameter that allows to estimate the frequency of cells in cell division, reflects cell proliferation and is regarded as an important parameter when determining the rate of plant root growth. Therefore, a low mitotic index (mitodepressive effect) characterize a few dividing cells [45-46] and is more frequently reported in the literature. This parameter predicts the root growth behavior, which is conducted by the frequency of cellular division in the root tissue. This index has been clearly correlated to the root length in the *A. cepa* test, as for instance decreasing in value with

Chromosomal aberration types detected may depend on DNA lesions and the cell cycle phase that is considered [46]. The mitotic anomalies often detected by the *Allium* test are: chromosome stickiness, multipolar anaphase, anaphasic bridge, c–mitosis or c–metaphase,

The induction of micronuclei in root meristems of *A. cepa* or in any cell of any other organism is the manifestation of chromosome damage and disturbance of the mitotic process. The micronucleus is formed by the development of a new membrane around the chromatin matter that failed to move to either pole during the anaphase of mitosis. Such chromatin matter arises either from anomalous disjunction of chromosomes due to spindle abnormalities or the breakage of chromosomes resulting in formation of acentric fragments, dicentric chromosomes and chromatin bridges. Thus, the induction of micronuclei may suggest that the environmental pollutant is either a spindle inhibitor or a clastogen. Induction of micronucleus formation, the outcome of chromosome breaks/fragments or

increasing concentrations of toxic metals like chromium and cadmium [46].

positive results on the environment and human health should be discussed.

induced mitotic spindle disturbance in *Allium cepa*.

**3.2. Evaluated parameters in** *Allium cepa* **test**

and micronuclei (Figure 2).

**Figure 2.** Mitotic anomalies and micronucleus observed in root meristematic cells of *Allium cepa* exposed to water samples of the Paraiba do Sul river - Brazil: a. chromosome bridges; b. not identified; c. multipolar anaphase; d. c-mitose; e. stickiness; f. micronuclei [47].

The use of *Allium cepa* as a test system was introduced by [23], when the effects of colchicine were investigated. This author defined it as an inactivation of segments achromatic spindle fibers by condensed chromosomes randomly dispersed throughout the cytoplasm cell. The formation of c-mitotic inhibitors relates to fibers of the spindle. Calmodulin, involved in regulating the concentration of calcium in the cell was specifically localized in the zone achromatic of mitotic cells, suggesting its involvement in the movement of chromosomes by controlling the polymerization and depolymerization of microtubules [50]. The storage irregular calcium in the cell inhibits the polymerization of microtubules, causing the formation of c-mitosis [51]. The c-mitosis may be defined, as well as the permanence of the equatorial plate anaphase chromosomes rather than move to their respective poles [21]. Cmitosis results from inactivation of the mitotic spindle followed by a random scattering of the condensed chromosomes [46].

The chromosome stickiness arises from improper folding of the chromosome fiber into single chromatids and chromosomes. As a result there is an intermingling of the fibers, and the chromosomes become attached to each other by means of subchromatid bridges. Chromosome stickiness has generally been inflicted by highly toxic agents, usually of an irreversible type, and probably leads cells to death [18,46]. Chromosome breaks - Any descriptive classification must include a category for the break or discontinuity, a simple severance of the chromosome or chromatid to give an acentric fragment, and which is not clearly associated with any exchange process [52]. Usually, the mitotic anomalie is the most frequently observed in studies conducted with *A. cepa*.

The chromosome bridges can involve one or more chromosomes. Bridges associated with stickiness promote a usually irreversible toxic effects.

The chromosome aberration terminology used in cytogenetic experiments generally comprehends chromosome changes designated as numerical (euploidy and aneuploidy) and structural (deletions, inversions, duplications, and translocations) aberrations. However, in experiments involving the *A. cepa* test, other parameters have also been considered to be important tools to inform on chromosome abnormalities as induced by cytotoxic or genotoxic agents. In this respect, we consider the term mitotic anomalies more appropriate for chromosomal changes observed in the *Allium* test [46].

Bioassays with Plants in the Monitoring of Water Quality 327

studies to evaluate the potential damage inflicted by pollutants in aquatic environments. Furthermore, even one bulb and one root per bulb was sufficient in discerning this damage, thereby shortening the time required to attain a statistically confident comparative evaluation. However, to allow for the use of statistical programs based on the evaluation of average values, and to avoid criticism based on genetic variability, we propose that three bulbs and

**Figure 3.** *Allium cepa* bioassay: a 20 L bucket was used for collecting the samples and transferred to 40 L containers (up); bulbs were exposed to treatment for 24 hours; growth root and root stained with Schiff

**4. Biomonitoring in the Vale do Paraíba region, São Paulo state, Brazil**

The São Paulo state is the most developed area of Brazil, with a high level of industrialization, expanding urbanization and high demographic growth rates. Consequently, there is a trend towards the worsening of some situations, as regards the deficiency in several aspects, such as infrastructure (difficulties to accommodate a population that grows every year), basic sanitation, lack of hydric resources before the demand and damage to their quality, mainly resulting from the direct dumping of urban and industrial sewage, untreated or inadequately treated, in rivers, lakes and reservoirs, among others. The main source of water pollution in the state of São Paulo is the dumping of domestic and industrial effluents, as well as the diffuse load of urban and agricultural.

The Paraíba do Sul river basin is located in the country's key economic area, which highlights for the diversity of its industrial park, especially the aeronautical and automotive industries, paper and cellulose, chemical, mechanical, electrical and electronic, and

(down).

three roots per bulb be considered as standard sample sizes for the *A. cepa* test [46].

#### **3.3. The** *Allium cepa* **protocol**

In the *Allium cepa* test protocol (based in Invittox Protocol n. 08 by [53]) for complex mixtures (as is the case of rivers and lakes waters), after having their ring of the root primordia carefully cleaned, the onion bulbs are exposed to clean water (it may be good quality tap water, Milli-Q water, distilled water, Hoagland solution, or mineral water, etc) for 48 hours in order to allow for the rooting of the bulb. After this period, bulbs were exposed to treatment for 24 hours: solution or water which needs to be tested, a negative control (no effect on the cell samples; the same solution for rooting is usually used) and a positive control (a drug used to promote the formation of abnormalities). A 20 L bucket was used for collecting the samples. The water was collected at bridges over the River passing alongside the mentioned cities (Figure 3). A mixture was composed from water sampled from the margins and the middle of the River, and transferred to 40 L containers.

All root tips are fixed in absolute ethanol-glacial acetic acid 3:1 (volume/volume) for 5 minutes and subjected to the Feulgen reaction en bloc. The acid hydrolysis pertinent to the Feulgen reaction is carried out in 4 M HCl at 24 °C for 75 minutes. Each stained root was squashed between slide and coverslip, and the squashes frozen in liquid nitrogen for the coverslip removal, and air dried. The preparations were then counterstained with Fast Green at pH 2.7 [54], rinsed in distilled water, air dried, cleared in xylene, and mounted in Canada balsam (Figure 3).

Although the *Allium cepa* test has been widely used to identify potentially cytotoxic and genotoxic pollutants in aquatic environments, variable non-standardized choices have been made regarding the number of plant bulbs and roots analyzed. We propose numbers for bulbs and roots per bulb when comparing the frequencies of micronuclei and mitotic anomalies with this test. When comparing quantitative biological data, one of the most frequent issues is the selection of an adequate number of samples. This decision is influenced by several factors, including the purpose of the study, the size of the population, the risk of using a sample that differs markedly from the population, and the sample error allowed. Determining the sample size is therefore of great importance; exceeding the necessary sample size may prove detrimental in terms of time and human and financial resources. However, too small of a sample size may lead to inconsistencies. As sample sizes increase, their variability tends to decrease, leading to a better hypothesis testing, a higher statistical power, and smaller confidence intervals. Nevertheless, there has not been a standardization of the ideal sample size of bulbs and their roots to be analyzed with the *A. cepa* test. The sampling of ten bulbs and five roots per bulb was adequate for comparative studies to evaluate the potential damage inflicted by pollutants in aquatic environments. Furthermore, even one bulb and one root per bulb was sufficient in discerning this damage, thereby shortening the time required to attain a statistically confident comparative evaluation. However, to allow for the use of statistical programs based on the evaluation of average values, and to avoid criticism based on genetic variability, we propose that three bulbs and three roots per bulb be considered as standard sample sizes for the *A. cepa* test [46].

326 Water Treatment

The chromosome aberration terminology used in cytogenetic experiments generally comprehends chromosome changes designated as numerical (euploidy and aneuploidy) and structural (deletions, inversions, duplications, and translocations) aberrations. However, in experiments involving the *A. cepa* test, other parameters have also been considered to be important tools to inform on chromosome abnormalities as induced by cytotoxic or genotoxic agents. In this respect, we consider the term mitotic anomalies more appropriate

In the *Allium cepa* test protocol (based in Invittox Protocol n. 08 by [53]) for complex mixtures (as is the case of rivers and lakes waters), after having their ring of the root primordia carefully cleaned, the onion bulbs are exposed to clean water (it may be good quality tap water, Milli-Q water, distilled water, Hoagland solution, or mineral water, etc) for 48 hours in order to allow for the rooting of the bulb. After this period, bulbs were exposed to treatment for 24 hours: solution or water which needs to be tested, a negative control (no effect on the cell samples; the same solution for rooting is usually used) and a positive control (a drug used to promote the formation of abnormalities). A 20 L bucket was used for collecting the samples. The water was collected at bridges over the River passing alongside the mentioned cities (Figure 3). A mixture was composed from water sampled

from the margins and the middle of the River, and transferred to 40 L containers.

All root tips are fixed in absolute ethanol-glacial acetic acid 3:1 (volume/volume) for 5 minutes and subjected to the Feulgen reaction en bloc. The acid hydrolysis pertinent to the Feulgen reaction is carried out in 4 M HCl at 24 °C for 75 minutes. Each stained root was squashed between slide and coverslip, and the squashes frozen in liquid nitrogen for the coverslip removal, and air dried. The preparations were then counterstained with Fast Green at pH 2.7 [54], rinsed in distilled water, air dried, cleared in xylene, and mounted in

Although the *Allium cepa* test has been widely used to identify potentially cytotoxic and genotoxic pollutants in aquatic environments, variable non-standardized choices have been made regarding the number of plant bulbs and roots analyzed. We propose numbers for bulbs and roots per bulb when comparing the frequencies of micronuclei and mitotic anomalies with this test. When comparing quantitative biological data, one of the most frequent issues is the selection of an adequate number of samples. This decision is influenced by several factors, including the purpose of the study, the size of the population, the risk of using a sample that differs markedly from the population, and the sample error allowed. Determining the sample size is therefore of great importance; exceeding the necessary sample size may prove detrimental in terms of time and human and financial resources. However, too small of a sample size may lead to inconsistencies. As sample sizes increase, their variability tends to decrease, leading to a better hypothesis testing, a higher statistical power, and smaller confidence intervals. Nevertheless, there has not been a standardization of the ideal sample size of bulbs and their roots to be analyzed with the *A. cepa* test. The sampling of ten bulbs and five roots per bulb was adequate for comparative

for chromosomal changes observed in the *Allium* test [46].

**3.3. The** *Allium cepa* **protocol**

Canada balsam (Figure 3).

**Figure 3.** *Allium cepa* bioassay: a 20 L bucket was used for collecting the samples and transferred to 40 L containers (up); bulbs were exposed to treatment for 24 hours; growth root and root stained with Schiff (down).

## **4. Biomonitoring in the Vale do Paraíba region, São Paulo state, Brazil**

The São Paulo state is the most developed area of Brazil, with a high level of industrialization, expanding urbanization and high demographic growth rates. Consequently, there is a trend towards the worsening of some situations, as regards the deficiency in several aspects, such as infrastructure (difficulties to accommodate a population that grows every year), basic sanitation, lack of hydric resources before the demand and damage to their quality, mainly resulting from the direct dumping of urban and industrial sewage, untreated or inadequately treated, in rivers, lakes and reservoirs, among others. The main source of water pollution in the state of São Paulo is the dumping of domestic and industrial effluents, as well as the diffuse load of urban and agricultural.

The Paraíba do Sul river basin is located in the country's key economic area, which highlights for the diversity of its industrial park, especially the aeronautical and automotive industries, paper and cellulose, chemical, mechanical, electrical and electronic, and extrativist industries, in addition to technological research centers. In agriculture, the predominance is of cultivations destined for animal production (cattle), extensive areas with the cultivation of eucalyptus, as well as the presence of the cultivations of rice, beans and corn. The basin draining area is about 55 500 km2, encompassing the states of São Paulo, Minas Gerais and Rio de Janeiro [47].

Bioassays with Plants in the Monitoring of Water Quality 329

activity. The reason for this contamination was, usually, the discharge of industrial effluents [55-56]. Other ecotoxicological tests, such as those conducted with *Ceriodaphnia dubia*, have shown, according to the CETESB 2007 Report, chronic toxicity in 21% of samples collected in

Considering the data involving root length growth and especially MI values, a cytotoxic potential is suggested for the water of the river Paraíba do Sul at Tremembé and Aparecida, in 2005. On the other hand, since in this year the MN frequency was not affected by the river water treatments, genotoxicity is not assumed for the river water sampled at the mentioned sites. By considering the frequencies of MN, CA and PI, a cytotoxic and genotoxic effect is supposed for *A. cepa* treated with the water collected from the river Paraíba do Sul at sites of Tremembé and Aparecida, in 2007. The *Allium* test has proved to be a sensitive tool for detecting cytotoxicity and genotoxicity effects promoted by the Paraíba do Sul water in the mentioned sites. Present findings reinforce the sensitivity of the *Allium cepa* test, especially concerning the MI evaluation, for monitoring river waters, thus serving as a tool for the early warning of the presence of cytotoxins in the hydric environment. We consider that this test could even be recommended for the prescreening of cytotoxicity in wastewaters [17,19]. In this study, MI and the MN frequency were determined by examination of 2,000 cells per slide, using 3-5 root tips from each bulb. The number of bulbs used depended on the availability of roots produced. The slides were examined from right to left, up end down, and the first 2,000 cells were scored for MI and MN frequency. Data on root length, MI, and the MN frequency were compared using ANOVA and the Tukey test. Variance heterogeneity was tested. Data on root length regarding the Tremembé site and the negative control in the year of 2005 were compared by a Welch test (t-test for different variances). The

Still in the year 2007, the genotoxic and citotoxic potential in water samples from the basin of Tapanhon river, Pindamonhangaba, São Paulo, using the *Allium* test was evaluated. Water samples of the Tapanhon river and its affluents (the Primeira Água and Segunda Água Streams, the Galega Brook) as well as a sample from the Borba Spring have been collected for negative control. Six bulbs of *Allium cepa* have been submitted to each of the samples for 24 hours. As such, 6000 cells have been collected for evaluation of the mitotic index (MI) and micronuclei (MNC) as well as 300 cells in the metaphase and anaphase phases, to evaluate chromosome aberrations (CA). No MNC has been found in the samples tested. The MI values have shown to be elevated in all samples tested when compared to the control with statistical relevance, however, only for the samples from the Primeira Água Stream and the Galega Brook (p<0,05). Chromosome aberrations have been observed in all samples (p<0,05). The averages of the parameters assessed were compared through ANOVA (0.05) of a factor, and the heteroscedasticity premise was assessed through the Levene test. The averages obtained were compared to one another through the Tukey test. The analyses

In 2008 a new study was carried out to characterize the mutagenic potential of the pollutants in the water of the Paraíba do Sul river, in Tremembé city, São Paulo State, Brazil, analyzing

analyses were done using Statistica and Statsdirect software [19].

were done using Statistica and Statsdirect software [25].

the Paraíba do Sul River.

The Paraíba do Sul river (Figure 4) is one of the components of the Paraíba do Sul basin, of importance to the southeast region of Brazil, as it serves for urban supply, irrigation, generation of energy, and assimilation of urban, industrial and agricultural discharges in the mentioned region. For this reason, since 2005, samples of water of this important river, collected in cities along its course, have been periodically evaluated. Anatomo-morphologic parameters (root growth), mitotic indices (MI), cell division phase indices (PI), frequency of micronuclei (MN) and chromosome anomalies (CA) are investigated.

**Figure 4.** Indication of the cities (Tremembé and Aparecida) 'triangles' at which the water was sampled from the Paraíba do Sul river [19].

The Environmental Sanitation and Technology Company (CETESB) performs the monitoring of the water bodies in the state of São Paulo, and has been trying to contribute with the pollution control and water quality recovery actions in the rivers and reservoirs in the state of São Paulo developed by the city, state and federal governments. In this monitoring, CETESB use 50 water quality variables (physical, chemical, hydrobiological, microbiological and ecotoxicology) [17]. In the last 20 years, 1007 surface water samples were analyzed through the Ames test at CETESB, and 137 (14%) of those showed mutagenic activity. The reason for this contamination was, usually, the discharge of industrial effluents [55-56]. Other ecotoxicological tests, such as those conducted with *Ceriodaphnia dubia*, have shown, according to the CETESB 2007 Report, chronic toxicity in 21% of samples collected in the Paraíba do Sul River.

328 Water Treatment

Minas Gerais and Rio de Janeiro [47].

from the Paraíba do Sul river [19].

extrativist industries, in addition to technological research centers. In agriculture, the predominance is of cultivations destined for animal production (cattle), extensive areas with the cultivation of eucalyptus, as well as the presence of the cultivations of rice, beans and corn. The basin draining area is about 55 500 km2, encompassing the states of São Paulo,

The Paraíba do Sul river (Figure 4) is one of the components of the Paraíba do Sul basin, of importance to the southeast region of Brazil, as it serves for urban supply, irrigation, generation of energy, and assimilation of urban, industrial and agricultural discharges in the mentioned region. For this reason, since 2005, samples of water of this important river, collected in cities along its course, have been periodically evaluated. Anatomo-morphologic parameters (root growth), mitotic indices (MI), cell division phase indices (PI), frequency of

**Figure 4.** Indication of the cities (Tremembé and Aparecida) 'triangles' at which the water was sampled

The Environmental Sanitation and Technology Company (CETESB) performs the monitoring of the water bodies in the state of São Paulo, and has been trying to contribute with the pollution control and water quality recovery actions in the rivers and reservoirs in the state of São Paulo developed by the city, state and federal governments. In this monitoring, CETESB use 50 water quality variables (physical, chemical, hydrobiological, microbiological and ecotoxicology) [17]. In the last 20 years, 1007 surface water samples were analyzed through the Ames test at CETESB, and 137 (14%) of those showed mutagenic

micronuclei (MN) and chromosome anomalies (CA) are investigated.

Considering the data involving root length growth and especially MI values, a cytotoxic potential is suggested for the water of the river Paraíba do Sul at Tremembé and Aparecida, in 2005. On the other hand, since in this year the MN frequency was not affected by the river water treatments, genotoxicity is not assumed for the river water sampled at the mentioned sites. By considering the frequencies of MN, CA and PI, a cytotoxic and genotoxic effect is supposed for *A. cepa* treated with the water collected from the river Paraíba do Sul at sites of Tremembé and Aparecida, in 2007. The *Allium* test has proved to be a sensitive tool for detecting cytotoxicity and genotoxicity effects promoted by the Paraíba do Sul water in the mentioned sites. Present findings reinforce the sensitivity of the *Allium cepa* test, especially concerning the MI evaluation, for monitoring river waters, thus serving as a tool for the early warning of the presence of cytotoxins in the hydric environment. We consider that this test could even be recommended for the prescreening of cytotoxicity in wastewaters [17,19]. In this study, MI and the MN frequency were determined by examination of 2,000 cells per slide, using 3-5 root tips from each bulb. The number of bulbs used depended on the availability of roots produced. The slides were examined from right to left, up end down, and the first 2,000 cells were scored for MI and MN frequency. Data on root length, MI, and the MN frequency were compared using ANOVA and the Tukey test. Variance heterogeneity was tested. Data on root length regarding the Tremembé site and the negative control in the year of 2005 were compared by a Welch test (t-test for different variances). The analyses were done using Statistica and Statsdirect software [19].

Still in the year 2007, the genotoxic and citotoxic potential in water samples from the basin of Tapanhon river, Pindamonhangaba, São Paulo, using the *Allium* test was evaluated. Water samples of the Tapanhon river and its affluents (the Primeira Água and Segunda Água Streams, the Galega Brook) as well as a sample from the Borba Spring have been collected for negative control. Six bulbs of *Allium cepa* have been submitted to each of the samples for 24 hours. As such, 6000 cells have been collected for evaluation of the mitotic index (MI) and micronuclei (MNC) as well as 300 cells in the metaphase and anaphase phases, to evaluate chromosome aberrations (CA). No MNC has been found in the samples tested. The MI values have shown to be elevated in all samples tested when compared to the control with statistical relevance, however, only for the samples from the Primeira Água Stream and the Galega Brook (p<0,05). Chromosome aberrations have been observed in all samples (p<0,05). The averages of the parameters assessed were compared through ANOVA (0.05) of a factor, and the heteroscedasticity premise was assessed through the Levene test. The averages obtained were compared to one another through the Tukey test. The analyses were done using Statistica and Statsdirect software [25].

In 2008 a new study was carried out to characterize the mutagenic potential of the pollutants in the water of the Paraíba do Sul river, in Tremembé city, São Paulo State, Brazil, analyzing

chromosomal changes in the meristematic cells of *Allium cepa*, in the summer (April) and winter (August). The bulbs were exposed for 72 h to the treatments: water from river, Hoagland solution (negative control) and 15 μg/L from MMS – methyl methanesulfonate (positive control). In each treatment, three bulbs were exposed and for each bulb, five slides were prepared. For mitotic index (MI) and micronucleus (MN) frequency rate, a total of 2,000 cells per root/slide were analyzed and 100 cells for the chromosome aberrations (CA). In April, the pollutants induced a high mutagenic potential in the meristematic cells of *Allium cepa*, the frequency rate of MN, stickiness and CA from non identified type were greater than the negative control. In August, the only significant change found was the chromosome bridges. There was no significant change for MI. These results bioindicator, therefore, it is important to keep biomonitoring and adopting effluents control measures.

Bioassays with Plants in the Monitoring of Water Quality 331

equipment and can be performed in partnership with teaching institutions, by providing specialized labor in the environmental area. In this sense, it is suggested to the official pollution control bodies that they implement the use of bioassays with plants as a screening before the expensive physical-chemical analyses, in order to spare time and money; and that partnerships with private initiative and teaching institutions can build on the relationship of the world with the quality of life throughout the terrestrial biome as regards the pollution

[1] World Health Organization – WHO (2012a) Health topics: water. Available:

[2] World Health Organization – WHO (2012b) Water scarcity. Available: http://www.who.int/features/factfiles/water/water\_facts/en/index1.html. Accessed 2012

[3] World Health Organization – WHO (2012c) Global Health Observatory: Water and sanitation. Available: http://www.who.int/gho/mdg/environmental\_sustainability/en/.

[4] WHO (1976) Surveillance of drinking-water quality. Geneva, World Health Organization. Available: http://whqlibdoc.who.int/monograph/WHO\_MONO\_63.pdf.

[5] World Health Organization – WHO (2011) Guidelines for drinking-water quality. WHO

http://www.who.int/water\_sanitation\_health/publications/2011/dwq\_guidelines/en/.

[6] Majer BJ, Grummt T, Uhl M, Knasmueller S (2005) Use of plant bioassays for the detection of genotoxins in the aquatic environment. Acta Hydrochim. Hydrobiol. 33: 45-

[7] Grant WF (1994) The present status of higher plant bioassays for the detection of

[8] Ma TH (1981) *Tradescantia* micronucleus bioassay and pollen tube chromatid aberration test for in situ monitoring and mutagen screening. Environ. Health Persp. 37: 85-90. [9] Ma TH (2001) *Tradescantia* micronucleo bioassay for detection of carcinogens. Folia

[10] Ma TH, Anderson VA, Harris MM, Neas RE, Lee TS (1985) Mutagenicity of drinking water detected by the *Tradescantia* micronucleus test. Can. J. Genet. Cytol. 27(2): 143-150.

http://www.who.int/topics/water/en/. Accessed 2012 Apr 10.

Library Cataloguing-in-Publication Data. Available:

environmental mutagens. Mutat. Res. 310: 175-185.

Histochem. Cytobiol. 39(Suppl. 2): 54-5.

control and environmental preservation.

*Institute of Bioscience, University of Taubaté, Brazil*

**Author details**

Agnes Barbério

**6. References**

Apr 11.

55.

Accessed 2012 Apr 10.

Accessed 2012 Feb 01.

Accessed 2012 Feb 01.

Macroscopic Parameters (root growth) were evaluated in the roots of 5 to 10 bulbs of *Allium cepa* submitted to samples of water of the Paraíba do Sul river collected in the city of Tremembé in August 2009 and 2010. Tap water was used as negative control. After 48 hours the roots of each bulb were measured in 72 hours (1st day) e 168 hours (5th day). There was no difference in the growth of roots in 2009, however, there was a difference in 2010, as well as between the years of 2009 and 2010 for the samples of the river. Several factors may have influenced this toxicity. August 2010 was the driest month in the state of São Paulo, with only 11 mm of rainfall registered. The low rainfall index may have promoted less dispersion of substances along the river. In the same period, the ecotoxicological result carried out by the CETESB using *Ceriodaphnia dubia* was chronic, what shows the presence of toxic substances present in water which inhibited the reproduction of this organism. The same toxicity was observed in our study for the decrease of the index of cellular division in the meristematic root of *A. cepa*.

Positive results, detected by the analysis of some parameters in bioassays with superior plants, indicate the presence of genotoxic and/or citotoxic substances in the environment, demonstrating a direct or indirect potential risk for living beings in contact with it.

Water is an essential resource to sustain life. As governments and community organizations make it a priority to deliver adequate supplies of quality water to people, individuals can help by learning how to conserve and protect the resource in their daily lives. In conclusion, for being an efficient and low cost, and easy to be executed tool, the *Allium* test is recommended for use in order to magnify the system of biomonitoring carried out by the inspection agencies.

#### **5. Conclusion**

The physical-chemical monitoring process is expensive, once it demands time, equipment, solutions and specialized labor. The biomonitoring techniques do not replace the assessments of the different parameters assessed through the conventional methods, but have great usefulness for a first diagnosis of the monitored and non-monitored areas, in order to effectively contribute in the pollution control process. The bioassays that use plants are effective, fast, easy to apply to any area, have low cost, do not need sophisticated equipment and can be performed in partnership with teaching institutions, by providing specialized labor in the environmental area. In this sense, it is suggested to the official pollution control bodies that they implement the use of bioassays with plants as a screening before the expensive physical-chemical analyses, in order to spare time and money; and that partnerships with private initiative and teaching institutions can build on the relationship of the world with the quality of life throughout the terrestrial biome as regards the pollution control and environmental preservation.

## **Author details**

330 Water Treatment

meristematic root of *A. cepa*.

inspection agencies.

**5. Conclusion**

chromosomal changes in the meristematic cells of *Allium cepa*, in the summer (April) and winter (August). The bulbs were exposed for 72 h to the treatments: water from river, Hoagland solution (negative control) and 15 μg/L from MMS – methyl methanesulfonate (positive control). In each treatment, three bulbs were exposed and for each bulb, five slides were prepared. For mitotic index (MI) and micronucleus (MN) frequency rate, a total of 2,000 cells per root/slide were analyzed and 100 cells for the chromosome aberrations (CA). In April, the pollutants induced a high mutagenic potential in the meristematic cells of *Allium cepa*, the frequency rate of MN, stickiness and CA from non identified type were greater than the negative control. In August, the only significant change found was the chromosome bridges. There was no significant change for MI. These results bioindicator, therefore, it is important to keep biomonitoring and adopting effluents control measures.

Macroscopic Parameters (root growth) were evaluated in the roots of 5 to 10 bulbs of *Allium cepa* submitted to samples of water of the Paraíba do Sul river collected in the city of Tremembé in August 2009 and 2010. Tap water was used as negative control. After 48 hours the roots of each bulb were measured in 72 hours (1st day) e 168 hours (5th day). There was no difference in the growth of roots in 2009, however, there was a difference in 2010, as well as between the years of 2009 and 2010 for the samples of the river. Several factors may have influenced this toxicity. August 2010 was the driest month in the state of São Paulo, with only 11 mm of rainfall registered. The low rainfall index may have promoted less dispersion of substances along the river. In the same period, the ecotoxicological result carried out by the CETESB using *Ceriodaphnia dubia* was chronic, what shows the presence of toxic substances present in water which inhibited the reproduction of this organism. The same toxicity was observed in our study for the decrease of the index of cellular division in the

Positive results, detected by the analysis of some parameters in bioassays with superior plants, indicate the presence of genotoxic and/or citotoxic substances in the environment,

Water is an essential resource to sustain life. As governments and community organizations make it a priority to deliver adequate supplies of quality water to people, individuals can help by learning how to conserve and protect the resource in their daily lives. In conclusion, for being an efficient and low cost, and easy to be executed tool, the *Allium* test is recommended for use in order to magnify the system of biomonitoring carried out by the

The physical-chemical monitoring process is expensive, once it demands time, equipment, solutions and specialized labor. The biomonitoring techniques do not replace the assessments of the different parameters assessed through the conventional methods, but have great usefulness for a first diagnosis of the monitored and non-monitored areas, in order to effectively contribute in the pollution control process. The bioassays that use plants are effective, fast, easy to apply to any area, have low cost, do not need sophisticated

demonstrating a direct or indirect potential risk for living beings in contact with it.

Agnes Barbério *Institute of Bioscience, University of Taubaté, Brazil*

## **6. References**

[11] Ruiz EF, Rabago VM, Lecona SU, Perez AB, Ma TH (1992) *Tradescantia* micronucleus (Trad-MCN) bioassay on clastogenicity of wastewater and in situ monitoring. Mutat. Res. 270(1): 45-51.

Bioassays with Plants in the Monitoring of Water Quality 333

[29] Barbosa JS, Cabral TM, Ferreira DN, Agnez-Lima LF, Batistuzzo De Medeiros SR (2010) Genotoxicity assessment in aquatic environment impacted by the presence of heavy

[30] Cotelle S, Masfaraud JF, Férard JF (1999) Assessment of the genotoxicity of contaminated soil with the *Allium*/*Vicia* micronucleus and the *Tradescantia* micronucleus

[31] Evseeva TI, Geras'kin SA, Shuktomova II (2003) Genotoxicity and toxicity assay of water sampled from a radium production industry storage cell territory by means of

[32] Palacio SM, Espinoza-Quiñones FR, Galante RM, Zenatti, DC, Seolatto AA, Lorenz EK, Zacarkin CE, Rossi N, Rizzutto MA, Tabacniks MH (2005) Correlation between heavy metal ions (Copper, Zinc, Lead) concentrations and root length of *Allium cepa* L. in

[33] Bortolotto T, Bertoldo JB, Silveira FZ, Defaveri TM, Silvano J, Pich CT (2009) Evaluation of the toxic and genotoxic potential of landfill leachates using bioassays. Environ.

[34] Radić S, Stipaničev D, Vujčić V, Rajčić MM, Širac S, Pevalek-Kozlina B (2010) The evaluation of surface and wastewater genotoxicity using the *Allium cepa* test. Sci. Total

[35] Cabrera GL, Rodriguez DMG (1999) Genotoxicity of leachates from a landfill using

[36] Cabrera GL, Rodriguez DMG, Maruri AB (1999) Genotoxicity of the extracts from the compost of the organic and the total municipal garbage using three plant bioassays.

[37] Monarca S, Feretti D, Collivignarelli C, Guzzella L, Zerbini I, Bertanza G, Pedrazzani R (2000) The influence of different disinfectants on mutagenicity and toxicity of urban

[38] Caritá R, Marin-Morales MA (2008) Induction of chromosome aberration in the *Allium cepa* test system caused by the exposure of seeds to industrial effluents contaminated

[39] Leme DM, De Angelis DF, Marin-Morales MA (2008) Action mechanisms of petroleum hydrocarbons present in waters impacted by an oil spill on the genetic material of

[40] Odeigah O, Nurudeen O, Amund OO (1997) Genotoxicity of oil field wastewater in

[41] Akintonwa A, Awodele O, Afolayan G, Coker HAB (2009) Mutagenic screening of some commonly used medicinal plants in Nigeria. J. Ethnopharmacol. 125: 461-470. [42] Çelik TA, Aslantürk OS (2010) Evaluation of cytotoxicity and genotoxicity of *Inula viscosa* leaf extracts with *Allium* test. J. Biom. Biotechnol. doi:10.1155/2010/189252 [43] Akinboro A, Bakare AA (2007) Cytotoxic and genotoxic effects of aqueous extracts of

five medicinal plants on *Allium cepa* Linn. J. Ethnopharmacol. 112: 470-475.

metals. Ecotoxicol. Environ. Saf. 73: 320-325.

*Allium* test. J. Environ. Radioact. 68: 235-248.

polluted river water. Braz. Arch. Biol. Technol. 48: 191-196.

assays. Mutat. Res. 426: 167-171.

Toxicol. Pharmacol. 28: 288-293.

three bioassays. Mutat. Res. 426: 207-210.

wastewater. Wat. Res. 34(17): 4261-4269.

with azo dyes. Chemosphere 72: 722-725.

Nigeria. Hereditas 126:161-167.

*Allium cepa* root cells. Aquat. Toxicol. 88: 214-219.

Environ. 408: 1228-1233.

Mutat. Res. 426: 201-206.

[29] Barbosa JS, Cabral TM, Ferreira DN, Agnez-Lima LF, Batistuzzo De Medeiros SR (2010) Genotoxicity assessment in aquatic environment impacted by the presence of heavy metals. Ecotoxicol. Environ. Saf. 73: 320-325.

332 Water Treatment

Res. 270(1): 45-51.

bioassay. Gen. Mol. Biol. 34(4): 689-693.

[11] Ruiz EF, Rabago VM, Lecona SU, Perez AB, Ma TH (1992) *Tradescantia* micronucleus (Trad-MCN) bioassay on clastogenicity of wastewater and in situ monitoring. Mutat.

[12] Thewes MR, Junior DE, Droste A (2011) Genotoxicity biomonitoring of sewage in two municipal wastewater treatment plants using the *Tradescantia pallida var. purpure*a

[13] Ma TH (1982) *Vicia* cytogenetic tests for environmental mutagens: A report of the U.S. environmental protection agency Gene-Tox program. Mutat. Res. 99(3): 257-271. [14] Kanaya N, Gill BS, Grover IS, Murin A, Osiecka R, Sandhu SS, Andersson HC (1994)

[15] Miao M, Fu R, Yang D, Zheng L (1999) *Vicia* root micronucleus assay on the clastogenicity of water samples from the Xiaoqing River in Shandong Province of the

[16] Grant WF, Owens ET (2006) *Zea mays* assays of chemical/radiation genotoxicity for the

[17] Barbério A (2009) Efeitos citotóxicos e genotóxicos no meristema radicular de *Allium cepa* exposta à água do rio Paraíba do Sul – estado de São Paulo – regiões de Tremembé

[18] Grant WF (1978) Chromosome aberrations in plants as a monitoring system. Environ.

[19] Barbério A, Barros L, Voltolini JC, Mello MLS (2009) Evaluation of the cytotoxic and genotoxic potential of water from the Brazilian river Paraíba do Sul with the *Allium cepa*

[20] Fiskesjö G (1988) The *Allium* test – an alternative in environmental studies: the relative

[21] Fiskesjö G (1985) The *Allium* test a standard in environmental monitoring. Hereditas

[22] Ma TH, Xu Z, Xu C, McConnell H, Rabago EV, Arreola GA, Zhang H (1995) The improved *Allium*/*Vicia* root tip micronucleus assay for clastogenicity of environmental

[23] Levan A (1938) Effect of colchicines on root mitosis in *Allium*. Hereditas 24(1): 471-486. [24] Grant WF (1982) Chromosome aberration assays in *Allium*: A report of the U. S. Environmental Protection Agency Gene-Tox Program. Mutat. Res. 99(3): 273-291. [25] Amaral AM, Barbério A, Voltolini JC, Barros L (2007) Avaliação preliminar da citotoxicidade e genotoxicidade da água na bacia do rio Tapanhon (SP-Brasil) através

[26] Grisolia CK, Oliveira ABB, Bonfim H (2005) Genotoxicity evaluation of domestic sewage in a municipal wastewater treatment plant. Gen. Mol. Biol. 28(2): 334 -338. [27] Bakare AA, Okunola AA, Adetunji OA, Jenmi HB (2009) Genotoxicity assessment of a

[28] Muzio MPH, Mendelson A, Magdaleno A, Tornello C, Balbis N, Moretton J (2006) Evaluation of genotoxicity and toxicity of Buenos Aires city hospital wastewater

pharmaceutical effluent using four bioassays. Gen. Mol. Biol. 32(2): 373-38.

do teste *Allium* (*Allium cepa*). Rev. Bras. Toxicol. 20(1-2): 65-71.

Vicia faba chromosomal aberration assay. Mutat. Res. 310(2): 231-247.

People's Republic of China. Mutat. Res. 426(2): 143-5.

e Aparecida. PhD thesis, Unicamp, Campinas.

toxicity of metal ions. Mutat. Res. 197(2): 243-260.

Health Perspect. 99: 273-291.

test. Braz. J. Biol. 69(3): 837-842.

pollutants. Mutat. Res. 334(2): 185-195.

samples. J. Braz. Soc. Ecotoxicol. 1(1): 1-6.

102(1): 99-112.

study of environmental mutagens. Mutat. Res. 613: 17-64.

[44] Egito LCM, Medeiros MG, De Medeiros SRB, Agnez-Lima LF (2007) Cytotoxic and genotoxic potential of surface water from the Pitimbu river, northeastern/RN Brazil. Genet. Mol. Biol. 30(2): 435-441.

**Chapter 14**

© 2013 Tahir Alp et al., licensee InTech. This is an open access chapter distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

© 2013 Tahir Alp et al., licensee InTech. This is a paper distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

**Relationship of Algae to Water Pollution and**

Pollution of surface water has become one of the most important environmental problems. Two types of large and long-lasting pollution threats can be recognized at the global level: on the one hand, organic pollution leading to high organic content in aquatic ecosystems and, in the long term, to eutrophication. It is a well-known fact that polluted water can

Organic pollution occurs when large quantities of organic compounds from many sources are released into the receiving running waters, lakes and also seas. Organic pollutants originate from domestic sewage (raw or treated), or urban run off, industrial effluents and farm water. Organic pollution could negatively affect the water quality in many ways. During the decomposition process of organic water dissolved oxygen in the water may be used up greater rate than it can be replenished thus, giving rise to oxygen depletion which causes severe consequences on the aquatic biota. Organic effluents also frequently contain large quantities of suspended solid which reduce the light available to photosynthetic organisms mainly algae. In addition organic wastes from people and animals may also rich

Algae are the main the primary producers in all kinds of water bodies and they are involved in water pollution in a number of significant ways. Firstly, enrichments of the algal nutrients in water through organic effluents may selectively stimulate the growth of algal species producing massive surface growths or 'blooms' that in turn reduce the water quality and affect its use. However, certain algae flourished in water polluted with organic wastes play an important part in "self-purification of water bodies". Some pollution algae may

reduce water quality thus restricting use of water bodies for many purposes.

**Waste Water Treatment**

Bulent Sen, Mehmet Tahir Alp, Feray Sonmez, Mehmet Ali Turan Kocer and Ozgur Canpolat

Additional information is available at the end of the chapter

in disease causing (pathogenic) organisms [1,2,3].

**2. Algae and water pollution**

http://dx.doi.org/10.5772/51927

**1. Introduction**

## **Relationship of Algae to Water Pollution and Waste Water Treatment**

Bulent Sen, Mehmet Tahir Alp, Feray Sonmez, Mehmet Ali Turan Kocer and Ozgur Canpolat

Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/51927

## **1. Introduction**

334 Water Treatment

Genet. Mol. Biol. 30(2): 435-441.

*Allium cepa*. Ambi-Agua 6(1): 90-103.

assays. Mutat. Res. 426(2): 183-188.

changes. J. Med. Gen. 12: 103-122.

survey. Mutat. Res. 491(1-2): 119-126.

for the *Allium cepa* test. Ecotoxicol. 20(4): 927-935.

Qual. 8: 291-298.

Biol. 13(1): 1-6.

House.

1980. p. 57-58.

[44] Egito LCM, Medeiros MG, De Medeiros SRB, Agnez-Lima LF (2007) Cytotoxic and genotoxic potential of surface water from the Pitimbu river, northeastern/RN Brazil.

[45] Fiskesjö G (1993) The *Allium* test in wastewater monitoring. Environ. Toxicol. Water.

[46] Barbério A, Voltolini JC, Mello MLS (2011) Standardization of bulb and root numbers

[47] Oliveira LM Oliveira LM, Voltolini JC, Barbério A (2011) Potencial mutagênico dos poluentes na água do rio Paraíba do Sul em Tremembé, SP, Brasil, utilizando o teste

[48] Grover IS, Kaur S (1999) Genotoxicity of wastewater samples from sewage and industrial effluent detected by the *Allium* root anaphase aberration and micronucleus

[49] Natarajan AT, Boei JJ, Darroudi F, Van Diemen PC, Dulout F, Hande MP, Ramalho AT (1996) Current cytogenetics methods for detecting exposure and effects of mutagens

[50] Li JX, Sun DY (1991) A study on CaM distribution in cells of living things. Chin. J. Cell

[51] Liu D, Jiang W. Li M (1992) Effects of trivalent and hexavalent chromium on root

[52] Savage JRK. (1975) Classification and relationships of induced chromosomal structural

[53] Fiskesjö G (1989) Invittox Protocol nº 8 - *Allium* test. Nottingham: Russel and Burch

[54] Mello MLS, Vidal BC Práticas de biologia celular. São Paulo: Edgard Blücher/Funcamp,

[55] Umbuzeiro G de A, Roubicek DA, Sanchez PS, Sato MI (2001) The Salmonella mutagenicity assay in a surface water quality monitoring program based on a 20-year

[56] Oliveira DP, Carneiro PA, Rech CM, Zanoni MV, Claxton LD, Umbuzeiro GA (2006) Mutagenic compounds generated from the chlorination of disperse Azo-Dyes and their

presence in drinking water. Environ. Sci. Technol. 40(21): 6682-6689.

and carcinogens. Environ. Health Perspect. 104(Suppl.3): 445-448.

growth and cell division of *Allium cepa*. Hereditas 117: 23-29.

Pollution of surface water has become one of the most important environmental problems. Two types of large and long-lasting pollution threats can be recognized at the global level: on the one hand, organic pollution leading to high organic content in aquatic ecosystems and, in the long term, to eutrophication. It is a well-known fact that polluted water can reduce water quality thus restricting use of water bodies for many purposes.

Organic pollution occurs when large quantities of organic compounds from many sources are released into the receiving running waters, lakes and also seas. Organic pollutants originate from domestic sewage (raw or treated), or urban run off, industrial effluents and farm water. Organic pollution could negatively affect the water quality in many ways. During the decomposition process of organic water dissolved oxygen in the water may be used up greater rate than it can be replenished thus, giving rise to oxygen depletion which causes severe consequences on the aquatic biota. Organic effluents also frequently contain large quantities of suspended solid which reduce the light available to photosynthetic organisms mainly algae. In addition organic wastes from people and animals may also rich in disease causing (pathogenic) organisms [1,2,3].

## **2. Algae and water pollution**

Algae are the main the primary producers in all kinds of water bodies and they are involved in water pollution in a number of significant ways. Firstly, enrichments of the algal nutrients in water through organic effluents may selectively stimulate the growth of algal species producing massive surface growths or 'blooms' that in turn reduce the water quality and affect its use. However, certain algae flourished in water polluted with organic wastes play an important part in "self-purification of water bodies". Some pollution algae may

© 2013 Tahir Alp et al., licensee InTech. This is an open access chapter distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. © 2013 Tahir Alp et al., licensee InTech. This is a paper distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

frequently are toxic to fish and also mankind and animals using polluted water. In fact, algae can play significant part of food chain of aquatic life, thus whatever alters the number and kinds of algae strongly affects all organisms in the chain including fish.

Relationship of Algae to Water Pollution and Waste Water Treatment 337

*Euglena*, diatoms, *Navicula*, *Synedra* and blue- green algae *Oscillatoria* and *Phormidium* are emphasized to tolerate organic pollution [12]. At species level, *Euglena viridis* (Euglenophyta), *Nitzschia palea* (Bacillariophyta), *Oscillatoria limosa*, *O.tenuis*, *O.princeps* and *Phormidium uncinatum* (Cyanophyta) are reported to be present than any other species in

**Plate 1.** 1.Stephanodiscus hantzschii Grunow 2.Cyclotella comta (Ehrenberg) Kützing 3.Thalassiosira weissflogii (Grunow) G.Fryxel & Hasle 4.Aulacoseira distans (Ehrenberg) Simonsen 5.Cyclotella ocellata Pantocsek 6. C. kützingiana Thwaites 7.Cocconeis pediculus Ehrenberg 8.C.placentula Ehrenberg 9. Meridion circulare (Greville) C. Agardh 10. Diatoma vulgaris var. lineare Grunow 11. D. tenuis C.Agardh 12. Surirella ovalis Brébisson 13. S. ovata var. apiculata W. Smith 14. S. linearis W.

Smith 15. S. minuta Brébisson 16. Cymatopleura solea (Brébisson) W. Smith [14].

Algae are also known to be causes of tastes and odors in water [4]. In fact, a large number of algae are associated with tastes and odors that vary in type. Certain diatoms, blue-green algae and coloured flagellates (particularly Chrysophyta and Euglenophyta) are the best known algae to pose such problems in water supplies, but green algae may also be involved. Some algae produce an aromatic odor resembling to that of particular flowers or vegetables. In addition, a spicy, a fishy odor and a grassy odor can also be produced by odor algae [5,6].

## **3. Algae as bioindicators**

Bioindicator organisms can be used to identify and qualify the effects of pollutants on the environment. Bioindicators can tell us about the cumulative effects of different pollutants in the ecosystem and about how long a problem may persist. Although indicator organisms can be any biological species that defines a trait or characteristics of the environment, algae are known to be good indicators of pollution of many types for the following reasons.

Algae of many kinds are really good indicators of water quality and many lakes are characterized based on their dominant phytoplankton groups. Many desmids are known to be present in oligotrophic waters whilst a few species frequently occurs in eutrophic bodies of water [7]. Similarly, many blue-green algae occurs in nutrient-poor waters, while some grow well in organically polluted waters [8]. The ecosystem approach to water quality assessment also include diatom species and accociations used as indicators of organic pollution. Five algal species were selected as indicators of the degree of pollution in rivers in England. *Stigeoclonium tenue* is present at the down stream margin of the heavily polluted part of a river, *Nitzschia palea and Gomphonema parvulum* always appear to be dominant in the mild pollution zone whilst *Cocconeis* and *Chamaesiphon* are reported to occur in unpolluted parts of the stream or in repurified zone [9]. *Navicula accomoda* is stressed to be a good indicator of sewage/organic pollution as the species comfortably occur in the most heavily polluted zones in which other species can not occur. The same hold true for species and varietes of *Gomphonema* [10] which is commonly found in highly organically polluted water. *Amphora ovalis* and *Gyrosigma attenuatum* are also introduced as good examples of diatoms to be affected by high organic content of water [11].

A list of more than 850 algal taxa was published based on the reports of considerable number of authors. According to this list, many algal genera have species that grow well in water containing a high concentration of organic wastes. Green algae *Chlamydomonas*, *Euglena*, diatoms, *Navicula*, *Synedra* and blue- green algae *Oscillatoria* and *Phormidium* are emphasized to tolerate organic pollution [12]. At species level, *Euglena viridis* (Euglenophyta), *Nitzschia palea* (Bacillariophyta), *Oscillatoria limosa*, *O.tenuis*, *O.princeps* and *Phormidium uncinatum* (Cyanophyta) are reported to be present than any other species in

336 Water Treatment

**3. Algae as bioindicators**

easier to detect and sample.

particularly to organic pollution

 algae have wide temporal and spatial distribution. many algal species are avaliable all the year.

diatoms to be affected by high organic content of water [11].

 response quickly to the charges in the environment due to pollution. Algae are diverse group of organisms found in large quantities.

frequently are toxic to fish and also mankind and animals using polluted water. In fact, algae can play significant part of food chain of aquatic life, thus whatever alters the number

Algae are also known to be causes of tastes and odors in water [4]. In fact, a large number of algae are associated with tastes and odors that vary in type. Certain diatoms, blue-green algae and coloured flagellates (particularly Chrysophyta and Euglenophyta) are the best known algae to pose such problems in water supplies, but green algae may also be involved. Some algae produce an aromatic odor resembling to that of particular flowers or vegetables. In addition, a spicy, a fishy odor and a grassy odor can also be produced by odor algae [5,6].

Bioindicator organisms can be used to identify and qualify the effects of pollutants on the environment. Bioindicators can tell us about the cumulative effects of different pollutants in the ecosystem and about how long a problem may persist. Although indicator organisms can be any biological species that defines a trait or characteristics of the environment, algae are known to be good indicators of pollution of many types for the following reasons.

The presence of some algae are well correlated with particular type of pollution

Algae of many kinds are really good indicators of water quality and many lakes are characterized based on their dominant phytoplankton groups. Many desmids are known to be present in oligotrophic waters whilst a few species frequently occurs in eutrophic bodies of water [7]. Similarly, many blue-green algae occurs in nutrient-poor waters, while some grow well in organically polluted waters [8]. The ecosystem approach to water quality assessment also include diatom species and accociations used as indicators of organic pollution. Five algal species were selected as indicators of the degree of pollution in rivers in England. *Stigeoclonium tenue* is present at the down stream margin of the heavily polluted part of a river, *Nitzschia palea and Gomphonema parvulum* always appear to be dominant in the mild pollution zone whilst *Cocconeis* and *Chamaesiphon* are reported to occur in unpolluted parts of the stream or in repurified zone [9]. *Navicula accomoda* is stressed to be a good indicator of sewage/organic pollution as the species comfortably occur in the most heavily polluted zones in which other species can not occur. The same hold true for species and varietes of *Gomphonema* [10] which is commonly found in highly organically polluted water. *Amphora ovalis* and *Gyrosigma attenuatum* are also introduced as good examples of

A list of more than 850 algal taxa was published based on the reports of considerable number of authors. According to this list, many algal genera have species that grow well in water containing a high concentration of organic wastes. Green algae *Chlamydomonas*,

and kinds of algae strongly affects all organisms in the chain including fish.

**Plate 1.** 1.Stephanodiscus hantzschii Grunow 2.Cyclotella comta (Ehrenberg) Kützing 3.Thalassiosira weissflogii (Grunow) G.Fryxel & Hasle 4.Aulacoseira distans (Ehrenberg) Simonsen 5.Cyclotella ocellata Pantocsek 6. C. kützingiana Thwaites 7.Cocconeis pediculus Ehrenberg 8.C.placentula Ehrenberg 9. Meridion circulare (Greville) C. Agardh 10. Diatoma vulgaris var. lineare Grunow 11. D. tenuis C.Agardh 12. Surirella ovalis Brébisson 13. S. ovata var. apiculata W. Smith 14. S. linearis W. Smith 15. S. minuta Brébisson 16. Cymatopleura solea (Brébisson) W. Smith [14].

organically polluted waters [13]. Some diatom taxa in a stream polluted with the waste water of a slaugher house are given in Plate 1-4 [14].

Relationship of Algae to Water Pollution and Waste Water Treatment 339

**Plate 3.** 1. *Rhopalodia gibba* (Ehrenberg) Otto Müller 2-3. *Eucocconeis flexella* (Kützing) Meister 4. *Achnanthidium minutisimum* (Kützing) Czarnecki 5. *Eucocconeis quadratarea* (Østrup) Lange-Bertalot 6. *Achnanthes marginulata* Grunow 7. *Ulnaria delicatissima* var. *angustissima* (Grunow) M. Aboal & P. C. Silva 8. *U. acus* (Kützing) M. Aboal 9. *U. amphirhyncus* (Ehrenberg) Compère & Bukhtiyarova 10. *U.*

*danica* (Kützing) Compère & Bukhtiyarova 11. *U. biceps* (Kützing) P. Compère [14].

**Plate 2.** 1. *Pinnularia viridis* (Nitzsch) Ehrenberg 2. *P. biceps* W. Gregory 3. *Stauroneis phoenicenteron* (Nitzsch) Ehrenberg 4. *Pinnularia brebissonii* (Kützing) Rabenhorst 5. *Craticula ambigua* (Ehrenberg) D. G. Mann 6. *Pinnularia mesolepta* (Ehrenberg) W. Smith [14].

organically polluted waters [13]. Some diatom taxa in a stream polluted with the waste

**Plate 2.** 1. *Pinnularia viridis* (Nitzsch) Ehrenberg 2. *P. biceps* W. Gregory 3. *Stauroneis phoenicenteron* (Nitzsch) Ehrenberg 4. *Pinnularia brebissonii* (Kützing) Rabenhorst 5. *Craticula ambigua* (Ehrenberg) D. G.

Mann 6. *Pinnularia mesolepta* (Ehrenberg) W. Smith [14].

water of a slaugher house are given in Plate 1-4 [14].

**Plate 3.** 1. *Rhopalodia gibba* (Ehrenberg) Otto Müller 2-3. *Eucocconeis flexella* (Kützing) Meister 4. *Achnanthidium minutisimum* (Kützing) Czarnecki 5. *Eucocconeis quadratarea* (Østrup) Lange-Bertalot 6. *Achnanthes marginulata* Grunow 7. *Ulnaria delicatissima* var. *angustissima* (Grunow) M. Aboal & P. C. Silva 8. *U. acus* (Kützing) M. Aboal 9. *U. amphirhyncus* (Ehrenberg) Compère & Bukhtiyarova 10. *U. danica* (Kützing) Compère & Bukhtiyarova 11. *U. biceps* (Kützing) P. Compère [14].

Relationship of Algae to Water Pollution and Waste Water Treatment 341

Algae are also good indicators of clean water since many species occur insistently and predominately in the clean water zone of the streams. However it is more satisfactory to emphasize the presence or absence of several species of clean water algae rather than of any one species to define the clean water zone. Approximately 46 taxa has been announced as representatives of the clean water algae including many diatoms, several flagellates and certain green and blue-green algae [12]. However it is emphasized that minute flagellates are better indicators of clean water than many larger algae. A few of the clean water algae are planktonic whilst many are benthic, attached to substrata at the bottom or sides of the

There are many studies by various authors emphasizing the relationships of algae to clean water. A community composed of the diatom *Cocconeis* and the blue-green alga *Chamaesiphon* is claimed to be present in the portion of the stream which has returned to normal following purification of a polluted condition [9]. Kolkwitz listed 61 diatoms, 42 green algae, 41 pigmented flagellates, 23 blue-green algae, and 5 red algae as organisms of oligosaprobic and /or unpolluted zones and Lackey found 77 species of planktonic algae in the clean water portion of a small stream, 40 of which were absent in the polluted area[15,16]. The flagellates *Chromulina rosanoffi*, *Mallomonas caudata,* the green algae *Ulothrix zonata* and *Microspora amoena* are also reported as oligosaprobic zone organisms [17]. Two groups of algae, Cryptophyta and Chrysophyta, are reported to be indicators of clean and/or unpolluted water as the members of these algal groups tend to occur in abundance, oppositely reacting adversely to pollution [18]. The absence of blue-green algae was also

The classic scheme for the interpretation of streams ecological conditions based on the biota was first introduced by Kolkwitz and Marsson [20]. They defined five zones based on the degree of pollution and proposed the use of aquatic organisms as indicators of different pollution and/or recovery zones of rivers which were polluted with organic matter such as sewage. However more recently Werner proposed nine different zones in the saprobic system in a stream organically polluted [21]. Survey of the saprobic zones and the corresponding communities are given Table 1. Pollution zones proposed in that saprobient system were basically termed "**Coprozoic**" "**Polysaprobic**", "**Mesosaprobic**", "**Oligosaprobic**" and "**Katharobic**". Each zone was different in chemical and physical characteristics and containing characteristic species. He listed indicator species of these zones except the last

Polysaprobic zone was characterized by almost complete absence of algae except for bluegreen alga *Arthrospira* (*Spirulina*) *jenneri* and green alga *Euglena viridis*. Bacteria and Protozoa were the most common groups in this zone. The preponderance of blue-green algae (Cyanophyta) was characteristic of alfa-mesosaprobic zone while diatoms (Bacillariophyta) and green algae (Chlorophyta) were dominant organisms in beta-mesosaprobic zone. Peridiniales (Dinophyta) and Charales (Charophyta) occurred in any quantity only in the

running waters.

accepted an indication of clean water [19].

**4. Use of algae in saprobien system**

one which is infact clean water.

**Plate 4.** 1.*Fragilaria capucina* var. *vaucheriae* (Kützing) Lange-Bertalot 2.*Pseudostaurosira brevistrita* (Grunow) D.M.Williams & Round 3.*Staurosirella pinnata* (Ehrenberg) D.M.Williams & Round 4.*Gomphonema truncatum* Ehrenberg 5.*Rhoicosphenia abbreviata* (C. Agardh) Lange-Bertalot 6.*Gomphonema olivaceum* (Hornemann) Brébisson 7.*Nitzschia sublinearis* Hustedt 8.N. *umbonata* (Ehrenberg) Lange-Bertalot 9. *N. hantzschiana* Rabenhorst 10. *Tryblionella angustata* W. Smith 11. *Nitzschia linearis* (C. Agardh) W. Smith12. *N. constricta* (Kützing) Ralfs. [14].

Algae are also good indicators of clean water since many species occur insistently and predominately in the clean water zone of the streams. However it is more satisfactory to emphasize the presence or absence of several species of clean water algae rather than of any one species to define the clean water zone. Approximately 46 taxa has been announced as representatives of the clean water algae including many diatoms, several flagellates and certain green and blue-green algae [12]. However it is emphasized that minute flagellates are better indicators of clean water than many larger algae. A few of the clean water algae are planktonic whilst many are benthic, attached to substrata at the bottom or sides of the running waters.

There are many studies by various authors emphasizing the relationships of algae to clean water. A community composed of the diatom *Cocconeis* and the blue-green alga *Chamaesiphon* is claimed to be present in the portion of the stream which has returned to normal following purification of a polluted condition [9]. Kolkwitz listed 61 diatoms, 42 green algae, 41 pigmented flagellates, 23 blue-green algae, and 5 red algae as organisms of oligosaprobic and /or unpolluted zones and Lackey found 77 species of planktonic algae in the clean water portion of a small stream, 40 of which were absent in the polluted area[15,16]. The flagellates *Chromulina rosanoffi*, *Mallomonas caudata,* the green algae *Ulothrix zonata* and *Microspora amoena* are also reported as oligosaprobic zone organisms [17]. Two groups of algae, Cryptophyta and Chrysophyta, are reported to be indicators of clean and/or unpolluted water as the members of these algal groups tend to occur in abundance, oppositely reacting adversely to pollution [18]. The absence of blue-green algae was also accepted an indication of clean water [19].

## **4. Use of algae in saprobien system**

340 Water Treatment

**Plate 4.** 1.*Fragilaria capucina* var. *vaucheriae* (Kützing) Lange-Bertalot 2.*Pseudostaurosira brevistrita* (Grunow) D.M.Williams & Round 3.*Staurosirella pinnata* (Ehrenberg) D.M.Williams & Round

Agardh) W. Smith12. *N. constricta* (Kützing) Ralfs. [14].

4.*Gomphonema truncatum* Ehrenberg 5.*Rhoicosphenia abbreviata* (C. Agardh) Lange-Bertalot 6.*Gomphonema olivaceum* (Hornemann) Brébisson 7.*Nitzschia sublinearis* Hustedt 8.N. *umbonata* (Ehrenberg) Lange-Bertalot 9. *N. hantzschiana* Rabenhorst 10. *Tryblionella angustata* W. Smith 11. *Nitzschia linearis* (C.

The classic scheme for the interpretation of streams ecological conditions based on the biota was first introduced by Kolkwitz and Marsson [20]. They defined five zones based on the degree of pollution and proposed the use of aquatic organisms as indicators of different pollution and/or recovery zones of rivers which were polluted with organic matter such as sewage. However more recently Werner proposed nine different zones in the saprobic system in a stream organically polluted [21]. Survey of the saprobic zones and the corresponding communities are given Table 1. Pollution zones proposed in that saprobient system were basically termed "**Coprozoic**" "**Polysaprobic**", "**Mesosaprobic**", "**Oligosaprobic**" and "**Katharobic**". Each zone was different in chemical and physical characteristics and containing characteristic species. He listed indicator species of these zones except the last one which is infact clean water.

Polysaprobic zone was characterized by almost complete absence of algae except for bluegreen alga *Arthrospira* (*Spirulina*) *jenneri* and green alga *Euglena viridis*. Bacteria and Protozoa were the most common groups in this zone. The preponderance of blue-green algae (Cyanophyta) was characteristic of alfa-mesosaprobic zone while diatoms (Bacillariophyta) and green algae (Chlorophyta) were dominant organisms in beta-mesosaprobic zone. Peridiniales (Dinophyta) and Charales (Charophyta) occurred in any quantity only in the

oligosaprobic zone. In the same zone, the bacterial count was low but there was a great variety of plants and animals (including fish) in considerable numbers.

Relationship of Algae to Water Pollution and Waste Water Treatment 343

**Zone I. Coprozoic zone**

**Zone II. α-Polysaprobic zone**

**Zone III. β-Polysaprobic zone**

**Zone IV. γ-Polysaprobic zone**

**Zone V. δ-Mesosaprobic zone**

**Zone VI. β-Mesosaprobic zone**

**Zone VII. γ-Mesosaprobic zone**

**Zone VIII. Oligosaprobic zone**

**Zone IX. Katharobic zone**

and the corresponding communities [21].

**a, b, c, d, e 1, 2, 3**

the bacterium community the *Bodo* community both communities

Rhodo-Thio bacterium community Pure Chlorobacterium community

Oscillatoria chlorina community Sphaerotilus natans community

Stigeoclonium tenue community

Cladophora fracta community Phormidium community

Lemanea fluviatilis)

(clean-water type))

Draparnaldia plumosa)

*Calothrix* species )

= as differences in degree

= as alternatives

Oscillatoria benthonicum community (O.brevis, O.limnosa, O.splendida with O.subtilissima, O. princeps and O.tenuis present as associate species)

Rhodophyceae community (Batrachospermum moniliforme or

Chlorophyceae community (*Draparnaldia glomerata*)

Chlorophyceae community (Chlorotylium cataractum and

Rhodophyceae community (*Hildenbrandia rivularis*)

**Table 1.** Aquatic communities representing various zones of pollution. Survey of the saprobic zones

Pure Meridion circulare community

vagum or Hildenbrandia rivularis) Vaucheria sessiis community Phormidium inundatum community

Chlorophyceae community (*Cladophora glomerata or Ulothrix zonata*

Rhodophyceae community (Lemanea annulata, Batrahcospermum

Lime-incrusting algal communities (*Chamaesiphon polonius* and various

Ulothrix zonata community

*Euglena* community

Beggiatoa community Thiothrix nivea mommunity

*Euglena* community

## **5. Use of algae in wastewater treatment**

Recently, algae have become significant organisms for biological purification of wastewater since they are able to accumulate plant nutrients, heavy metals, pesticides, organic and inorganic toxic substances and radioactive matters in their cells/bodies [22-25]. Biological wastewater treatment systems with micro algae have particularly gained importance in last 50 years and it is now widely accepted that algal wastewater treatment systems are as effective as conventional treatment systems. These spesific features have made algal wastewaters treatment systems an significant low-cost alternatives to complex expensive treatment systems particularly for purification of municipal wastewaters.

In addition, algae harvested from treatment ponds are widely used as nitrogen and phophorus suplement for agricultural purpose and can be subjected to fermentation in order to obtain energy from metane. Algae are also able to accumulate highly toxic substances such as selenium, zinc and arsenic in their cells and/or bodies thus eliminating such substances from aquatic enviroments. Radiation is also an important type of pollution as some water contain naturally radioactive materials, and others become radioactive through contamination. Many algae can take up and accumulate many radioactive minerals in their cells even from greater concentrations in the water [12]. MacKenthun emphasized that *Spirogyra* can accumulate radio-phosphorus by a factor 850.000 times that of water [26]. Considering all these abilities of algae to purify the polluted waters of many types, it is worth to emphasize that algal technology in wastewater treatment systems are expected to get even more common in future years.

Wastewater treatment which is applied to improve or upgrade the quality of a wastewater involves physical, chemical and biological processes in primary, secondary or tertiary stages. Primary treatment removes materials that will either float or readily settle out by gravity. It includes the physical processes of screening, commination, grit removal, and sedimentation. While the secondary treatment is usually accomplished by biological processes and removes the soluble organic matter and suspended solids left from primary treatment. Tertiary or advanced treatment is process for purification in which nitrates and phosphates, as well as fine particles are removed [27]. However initial cost as well as operating cost of wastewater treatment plant including primary, secondary or advanced stages is highly expensive [28].

It is well known that algae have an important role in self purification of organic pollution in natural waters [29]. Moreover, many studies revealed that algae remove nutrients especially nitrogen and phosphorus, heavy metals, pesticides, organic and inorganic toxins, pathogens from surrounding water by accumulating and/or using them in their cells [30,31,32,33,34,35,36,37,38]. Also, studies showed that algae may be used successfully for wastewater treatment as a result of their bioaccumulation abilities [39].

oligosaprobic zone. In the same zone, the bacterial count was low but there was a great

Recently, algae have become significant organisms for biological purification of wastewater since they are able to accumulate plant nutrients, heavy metals, pesticides, organic and inorganic toxic substances and radioactive matters in their cells/bodies [22-25]. Biological wastewater treatment systems with micro algae have particularly gained importance in last 50 years and it is now widely accepted that algal wastewater treatment systems are as effective as conventional treatment systems. These spesific features have made algal wastewaters treatment systems an significant low-cost alternatives to complex expensive

In addition, algae harvested from treatment ponds are widely used as nitrogen and phophorus suplement for agricultural purpose and can be subjected to fermentation in order to obtain energy from metane. Algae are also able to accumulate highly toxic substances such as selenium, zinc and arsenic in their cells and/or bodies thus eliminating such substances from aquatic enviroments. Radiation is also an important type of pollution as some water contain naturally radioactive materials, and others become radioactive through contamination. Many algae can take up and accumulate many radioactive minerals in their cells even from greater concentrations in the water [12]. MacKenthun emphasized that *Spirogyra* can accumulate radio-phosphorus by a factor 850.000 times that of water [26]. Considering all these abilities of algae to purify the polluted waters of many types, it is worth to emphasize that algal technology in wastewater treatment systems are expected to

Wastewater treatment which is applied to improve or upgrade the quality of a wastewater involves physical, chemical and biological processes in primary, secondary or tertiary stages. Primary treatment removes materials that will either float or readily settle out by gravity. It includes the physical processes of screening, commination, grit removal, and sedimentation. While the secondary treatment is usually accomplished by biological processes and removes the soluble organic matter and suspended solids left from primary treatment. Tertiary or advanced treatment is process for purification in which nitrates and phosphates, as well as fine particles are removed [27]. However initial cost as well as operating cost of wastewater treatment plant including primary, secondary or advanced

It is well known that algae have an important role in self purification of organic pollution in natural waters [29]. Moreover, many studies revealed that algae remove nutrients especially nitrogen and phosphorus, heavy metals, pesticides, organic and inorganic toxins, pathogens from surrounding water by accumulating and/or using them in their cells [30,31,32,33,34,35,36,37,38]. Also, studies showed that algae may be used successfully for

wastewater treatment as a result of their bioaccumulation abilities [39].

variety of plants and animals (including fish) in considerable numbers.

treatment systems particularly for purification of municipal wastewaters.

**5. Use of algae in wastewater treatment**

get even more common in future years.

stages is highly expensive [28].

**Table 1.** Aquatic communities representing various zones of pollution. Survey of the saprobic zones and the corresponding communities [21].

## **6. Advantages of use of algae in wastewater treatment**

There are a symbiotic relation among bacteria and algae in aquatic ecosystems. Algae support to aerobic bacterial oxidation of organic matter producing oxygen via photosynthesis whilst released carbon dioxide and nutrients in aerobic oxidation use for growth of algal biomass. Considering ammonium, carbon dioxide and orthophosphate as main nutrient sources, Oswald determined that oxygen release ratio is 1.5 g O2/1 g algal biomass [40]. Grobbelaar et al. reported to oxygen release ratio of 1.9 g O2/1 g algal biomass [41]. Arceivala, accounting latitude, climate and atmospheric conditions, calculated that 4- 6% of mean daily solar radiation reaching on treatment pond in 40°N latitude use for new biomass production and production rate of algal biomass may reach 80 kg O2/1 ha-day [42].

Relationship of Algae to Water Pollution and Waste Water Treatment 345

**Figure 1.** Removal efficiency of organic nitrogen in treatment methods [44]. 1-nitrification, 2-oxİdation pond, 3-chemical coagulation, 4-chlorination, 5-ammonia removal, 6-filtration, 7-reverse osmosis

Facultative ponds (usually 2.5m in depth) are systems where effluent quality improves between 5 and 30 days depending on factors such as climate, temperature, wind and surface area [52]. There are three main zones in such ponds; two upper zones with oxygen whilst anaerobic conditions prevail in bottom. Algal photosynthesis and atmospheric diffusion are main oxygen source. Wastes are stabilized by aerobic bacteria in upper zone and by facultative bacteria in intermediate depths while degraded by anaerobic bacteria in bottom zone [53]. Zooplankton controls to excessive bacterial growth and algal blooms through grazing as well as contributing to carbon dioxide production for algal

Acceptable effluent quality is the most important advantage of facultative ponds though low operation and maintenance costs. However there are some disadvantages such as high land costs, odour problem in high waste loading, loss of nitrogen to atmosphere, limiting the nutrient reuse by phosphate sedimentation also limiting of irrigation potential by salinity increase during high evaporation period [54]. Although temperature largely affects retention time of wastewater, facultative ponds are widely used in different climate regimes. For example there are more than 3000 facultative ponds in Germany and France and 7000 in

photosynthesis. Food web in a facultative pound is given in Figure 2.

United States [53].

Most of nitrogen in algal cell bound to proteins which compose to 45-60% of dry weight and phosphorus is essential for synthesis of nucleic acids, phospholipids and phosphate esters. Algae using nitrogen and phosphorus in growth may remove to nutrients load of wastewater from a few hours to a few days [43].

In comparison to common treatment systems, oxidation ponds supporting growth of some species may be effective of nutrient removal (Fig. 1). Increasing dissolved oxygen concentration and pH cause for phosphorus sedimentation, ammonia and hydrogen sulphur removal. High pH in algal ponds also leads to pathogen disinfection [44]. Removal efficiency of heavy metals by algae shows changes among species. In fact, studies showed that chrome by *Oscillatoria*, cadmium, copper and zinc by *Chlorella vulgaris*, lead by *Chlamydomonas* and molybdenum by *Scenedesmus chlorelloides* may remove successfully [45,46,47,48,49]. Although algae have adaptation ability to sub-lethal concentrations, accumulation of heavy metals in cells may be potentially toxic effects to the other circles of food web [50].

## **7. Algal-bacterial ponds**

Algal-bacterial pond is water body which is designed to keep and improve of wastewater in a certain time. Although wastewater is treated in pond via physical, chemical and biological processes and/or mechanical processes like aeration, there are also ponds completely based on processes of natural conditions. Ponds, where stabilization of dissolved compounds and suspended solids is in completely aerobic conditions, are named "oxidation ponds". When stabilisation in anaerobic or facultative conditions, ponds are named "waste stabilisation ponds". Stabilisation pond systems are assessed in different types: facultative, anaerobic, aeration and maturation ponds. Common pond type which utilizes from algae is facultative stabilisation ponds. Facultative ponds are designed for purposes such as decrease of waste retention time, achieve of effective treatment or algal culture (Fig. 2). Algal photosynthesis and bacterial decomposition is principal mechanism of algal-bacterial ponds. The processes including oxidation, settling, sedimentation, adsorption, disinfection in the ponds are results of symbiotic relation between algae and bacteria populations [51].

food web [50].

**7. Algal-bacterial ponds**

**6. Advantages of use of algae in wastewater treatment**

wastewater from a few hours to a few days [43].

There are a symbiotic relation among bacteria and algae in aquatic ecosystems. Algae support to aerobic bacterial oxidation of organic matter producing oxygen via photosynthesis whilst released carbon dioxide and nutrients in aerobic oxidation use for growth of algal biomass. Considering ammonium, carbon dioxide and orthophosphate as main nutrient sources, Oswald determined that oxygen release ratio is 1.5 g O2/1 g algal biomass [40]. Grobbelaar et al. reported to oxygen release ratio of 1.9 g O2/1 g algal biomass [41]. Arceivala, accounting latitude, climate and atmospheric conditions, calculated that 4- 6% of mean daily solar radiation reaching on treatment pond in 40°N latitude use for new biomass production and production rate of algal biomass may reach 80 kg O2/1 ha-day [42].

Most of nitrogen in algal cell bound to proteins which compose to 45-60% of dry weight and phosphorus is essential for synthesis of nucleic acids, phospholipids and phosphate esters. Algae using nitrogen and phosphorus in growth may remove to nutrients load of

In comparison to common treatment systems, oxidation ponds supporting growth of some species may be effective of nutrient removal (Fig. 1). Increasing dissolved oxygen concentration and pH cause for phosphorus sedimentation, ammonia and hydrogen sulphur removal. High pH in algal ponds also leads to pathogen disinfection [44]. Removal efficiency of heavy metals by algae shows changes among species. In fact, studies showed that chrome by *Oscillatoria*, cadmium, copper and zinc by *Chlorella vulgaris*, lead by *Chlamydomonas* and molybdenum by *Scenedesmus chlorelloides* may remove successfully [45,46,47,48,49]. Although algae have adaptation ability to sub-lethal concentrations, accumulation of heavy metals in cells may be potentially toxic effects to the other circles of

Algal-bacterial pond is water body which is designed to keep and improve of wastewater in a certain time. Although wastewater is treated in pond via physical, chemical and biological processes and/or mechanical processes like aeration, there are also ponds completely based on processes of natural conditions. Ponds, where stabilization of dissolved compounds and suspended solids is in completely aerobic conditions, are named "oxidation ponds". When stabilisation in anaerobic or facultative conditions, ponds are named "waste stabilisation ponds". Stabilisation pond systems are assessed in different types: facultative, anaerobic, aeration and maturation ponds. Common pond type which utilizes from algae is facultative stabilisation ponds. Facultative ponds are designed for purposes such as decrease of waste retention time, achieve of effective treatment or algal culture (Fig. 2). Algal photosynthesis and bacterial decomposition is principal mechanism of algal-bacterial ponds. The processes including oxidation, settling, sedimentation, adsorption, disinfection in the ponds are results

of symbiotic relation between algae and bacteria populations [51].

**Figure 1.** Removal efficiency of organic nitrogen in treatment methods [44]. 1-nitrification, 2-oxİdation pond, 3-chemical coagulation, 4-chlorination, 5-ammonia removal, 6-filtration, 7-reverse osmosis

Facultative ponds (usually 2.5m in depth) are systems where effluent quality improves between 5 and 30 days depending on factors such as climate, temperature, wind and surface area [52]. There are three main zones in such ponds; two upper zones with oxygen whilst anaerobic conditions prevail in bottom. Algal photosynthesis and atmospheric diffusion are main oxygen source. Wastes are stabilized by aerobic bacteria in upper zone and by facultative bacteria in intermediate depths while degraded by anaerobic bacteria in bottom zone [53]. Zooplankton controls to excessive bacterial growth and algal blooms through grazing as well as contributing to carbon dioxide production for algal photosynthesis. Food web in a facultative pound is given in Figure 2.

Acceptable effluent quality is the most important advantage of facultative ponds though low operation and maintenance costs. However there are some disadvantages such as high land costs, odour problem in high waste loading, loss of nitrogen to atmosphere, limiting the nutrient reuse by phosphate sedimentation also limiting of irrigation potential by salinity increase during high evaporation period [54]. Although temperature largely affects retention time of wastewater, facultative ponds are widely used in different climate regimes. For example there are more than 3000 facultative ponds in Germany and France and 7000 in United States [53].

## **8. High rate algal ponds**

Municipal wastewater treatments with high rate algal ponds were first proposed by Oswald and Golouke and thereinafter were used in many parts of the world [55,56]. High rate algal pond is usually shallow (20-50 cm) and is equipped with mechanical aeration and mixing by means of paddle wheels. High oxygen level resulting from photosynthesis and aeration allows to low retention times in these ponds. Removal rates of high rate algal ponds are almost similar to conventional treatment methods but may also be more efficient with lower retention time. In fact biochemical oxygen demand (BOD) up to 90% and more than 80% of nitrogen and phosphorus are treated in high rate algal ponds in a few days. However required time for treatment of biochemical oxygen demand up to 90% using by conventional activated sludge and bio filtration techniques, which are highly expensive secondary treatment methods, is between five and eight hours during which lower ratio of nitrogen and phosphorus may be removed. Further, construction and energy costs are highly lower and land requirement is half the required for facultative ponds [57]. It is a well-known fact that only a small amount of nitrogen and phosphorus are removed in active sludge and bio filtration techniques, In addition active sludge and bio filtration techniques require expensive chemicals and complex systems.

Relationship of Algae to Water Pollution and Waste Water Treatment 347

The total pond area needed is much larger than that needed for a conventional plant, but ponds should still cost only one-third to one-half as much to build. Another important advantage of the plant is the small amount of sludge they produced. For example, during nearly 3 decades of operation, St. Helena's wastewater treatment plant has never had to remove residue and measured less than 1 meter of residue had accumulated at the bottom of

the deep digester pit [59].

**Figure 2.** Food web in facultative wastewater treatment pond [51]

Cost of harvest in high rate algal ponds may be most important problem. Thus sedimentation of algae with flocculating is aimed when the wheels are stopped for harvest. In addition growth of resistant algal species to sinking such as *Chlorella, Euglena, Chlamydomonas* and *Oscillatoria* is undesired algae in the ponds. *Scenedesmus* or *Micractenium*, non-preferred species due to their cell morphology for grazing, are dominant in well mixed ponds [40]. Harvested algae may use for industrial and agricultural use as well as effluent in aquaculture (Fig. 3).

## **9.  The first pond in series is a facultative pond with depth of 4 to 5 m containing a digester pit, which functions much like an anaerobic pond while surface zone remains aerobic. Effluent of the facultative pond flows to the high rate algal pond for remove to dissolved organic matter and nutrients, then to settling pond with residence time of one or two days for sedimentation of algae and suspended solids. The last unit is maturation ponds where treated water is exposed to the sun and wind leading to natural oxygenation and solar disinfection, and thus an inactivation of pathogens [58].

Wastewater Treatment and Reclamation Plant in St. Helena, California, by US Department of Energy built of formed earth rather than of reinforced concrete in the early 1960s (Fig. 4). The total pond area needed is much larger than that needed for a conventional plant, but ponds should still cost only one-third to one-half as much to build. Another important advantage of the plant is the small amount of sludge they produced. For example, during nearly 3 decades of operation, St. Helena's wastewater treatment plant has never had to remove residue and measured less than 1 meter of residue had accumulated at the bottom of the deep digester pit [59].

346 Water Treatment

**8. High rate algal ponds**

expensive chemicals and complex systems.

**9. Advanced integrated wastewater ponds**

aquaculture (Fig. 3).

pathogens [58].

Municipal wastewater treatments with high rate algal ponds were first proposed by Oswald and Golouke and thereinafter were used in many parts of the world [55,56]. High rate algal pond is usually shallow (20-50 cm) and is equipped with mechanical aeration and mixing by means of paddle wheels. High oxygen level resulting from photosynthesis and aeration allows to low retention times in these ponds. Removal rates of high rate algal ponds are almost similar to conventional treatment methods but may also be more efficient with lower retention time. In fact biochemical oxygen demand (BOD) up to 90% and more than 80% of nitrogen and phosphorus are treated in high rate algal ponds in a few days. However required time for treatment of biochemical oxygen demand up to 90% using by conventional activated sludge and bio filtration techniques, which are highly expensive secondary treatment methods, is between five and eight hours during which lower ratio of nitrogen and phosphorus may be removed. Further, construction and energy costs are highly lower and land requirement is half the required for facultative ponds [57]. It is a well-known fact that only a small amount of nitrogen and phosphorus are removed in active sludge and bio filtration techniques, In addition active sludge and bio filtration techniques require

Cost of harvest in high rate algal ponds may be most important problem. Thus sedimentation of algae with flocculating is aimed when the wheels are stopped for harvest. In addition growth of resistant algal species to sinking such as *Chlorella, Euglena, Chlamydomonas* and *Oscillatoria* is undesired algae in the ponds. *Scenedesmus* or *Micractenium*, non-preferred species due to their cell morphology for grazing, are dominant in well mixed ponds [40]. Harvested algae may use for industrial and agricultural use as well as effluent in

Advanced integrated wastewater pond systems are an adaptation of waste stabilisation ponds systems based on a series of four advanced ponds: A facultative pond; a high rate algal pond; an algal settling pond and finally a maturation pond for solar disinfection and pathogen abatement. The first pond in series is a facultative pond with depth of 4 to 5 m containing a digester pit, which functions much like an anaerobic pond while surface zone remains aerobic. Effluent of the facultative pond flows to the high rate algal pond for remove to dissolved organic matter and nutrients, then to settling pond with residence time of one or two days for sedimentation of algae and suspended solids. The last unit is maturation ponds where treated water is exposed to the sun and wind leading to natural oxygenation and solar disinfection, and thus an inactivation of

Wastewater Treatment and Reclamation Plant in St. Helena, California, by US Department of Energy built of formed earth rather than of reinforced concrete in the early 1960s (Fig. 4).

**Figure 2.** Food web in facultative wastewater treatment pond [51]

Relationship of Algae to Water Pollution and Waste Water Treatment 349

Pond Integrated Pond

cm) 4-5 m

O2 level is high) high

lower) high

High rate Algae

Criteria Bacteria-Algae

Operation/maintenance

**10. Conclusion**

cost

Pond

Salinity increase - -

Land required high low low Odor problem occur not occur not occur Loss of N to atmosphere occur not occur not occur

PO4 sedimantation occur not occur not occur Time required for treatment 5-30 days a few days 5-6 days Energy requirement low low high Efficiency quality low high high BOD removal fair good good Suspended solid removal fair good good Harvesting cost high low

The water flows from lands into aquatic environments contribute enormous amounts of organic matters and plants nutrients to the aquatic systems which give rise to eutrophication and pollution. With increased urbanization the need for sewage treatment plants (STP) became more important. Wastewater treatment which is applied to improve or upgrade the quality of a wastewater involves physical, chemical and biological processes in primary, secondary or tertiary stages.More sewage plants are designed to remove solids (primary process), followed by a secondary process which involves either activated sludge or trickling filters to reduce the Biological Oxygen Demand (BOD). Removal of the nutrients left after secondary treatment is possible by a variety of processes, one of which involves growth and harvesting of algae from the effluents: others involve ion exchance electro chemical, electrodialysis, reserve osmosis, distillation, chemical precipitation as tertiary processes. However initial cost as well as operating cost of wastewater treatment plant

low lower (5 folds

Depth 2.5 m 0.2-0.5 m (20-50

Retention time low lower (because

**Table 2.** Comparison of wastetreatment ponds in terms of various criteria

including primary, secondary or advanced stages is highly expensive.

**Figure 3.** Flow Scheme of the Accelerated Photosynthetic Process for Waste-Water Treatment and Algal Protein Production [56]

**Figure 4.** Diagram of St. Helena advanced integrated pond system [59]

Comparison of algae involved wastewater treatment systems is given at table 2. Many useful criteria have been used in the table for more constructive and trustable comparison of wastewater treatment systems connected to algae.

**Table 2.** Comparison of wastetreatment ponds in terms of various criteria

## **10. Conclusion**

348 Water Treatment

Protein Production [56]

**Figure 3.** Flow Scheme of the Accelerated Photosynthetic Process for Waste-Water Treatment and Algal

Comparison of algae involved wastewater treatment systems is given at table 2. Many useful criteria have been used in the table for more constructive and trustable comparison of

**Figure 4.** Diagram of St. Helena advanced integrated pond system [59]

wastewater treatment systems connected to algae.

The water flows from lands into aquatic environments contribute enormous amounts of organic matters and plants nutrients to the aquatic systems which give rise to eutrophication and pollution. With increased urbanization the need for sewage treatment plants (STP) became more important. Wastewater treatment which is applied to improve or upgrade the quality of a wastewater involves physical, chemical and biological processes in primary, secondary or tertiary stages.More sewage plants are designed to remove solids (primary process), followed by a secondary process which involves either activated sludge or trickling filters to reduce the Biological Oxygen Demand (BOD). Removal of the nutrients left after secondary treatment is possible by a variety of processes, one of which involves growth and harvesting of algae from the effluents: others involve ion exchance electro chemical, electrodialysis, reserve osmosis, distillation, chemical precipitation as tertiary processes. However initial cost as well as operating cost of wastewater treatment plant including primary, secondary or advanced stages is highly expensive.

Recently, algae have become significant organisms for biological purification of wastewater since they are able to accumulate plant nutrients, heavy metals, pesticides, organic and inorganic toxic substances and radioactive matters in their cells/bodies with their bioaccumulation abilities. Particularly, biological wastewater treatment systems with micro algae have gained great importance in last 50 years and it is now widely accepted that algal wastewater treatment systems are as effective as conventional treatment systems. Removal rates of particularly high rate algal ponds are almost similar to conventional treatment methods but it is more efficient with lower retention time. With these spesific features algal wastewater treatment systems can be accepted as an significant low-cost alternatives to complex expensive treatment systems particularly for purification of municipal wastewaters.

Relationship of Algae to Water Pollution and Waste Water Treatment 351

[7] Brook, A.J. Planktonic algae as indicators of lake types, with special reference to the

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[18] Lackey, J.B. Two groups of flagellated algae serving as indicators of clean water. J. Am.

[19] Rafter, G.W. The microscopical examination of potable water. Van Nostrand Co., N.Y.

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## **Author details**

Bulent Sen, Feray Sonmez and Ozgur Canpolat *University of Firat, Faculty of Fisheries Department of Aquatic Basic Sciences Elazğ, Turkey*

Mehmet Tahir Alp\* *University of Mersin, Faculty of Fisheries Department of Aquatic Basic Sciences Mersin, Turkey*

Mehmet Ali Turan Kocer *Mediterranean Fisheries Research Production and Training Institue, Antalya, Turkey*

## **11. References**

<sup>\*</sup> Corresponding Author

[7] Brook, A.J. Planktonic algae as indicators of lake types, with special reference to the Desmidiaceae. Limnol and Oceannog 1965;10; 403-411.

350 Water Treatment

wastewaters.

**Author details**

Mehmet Tahir Alp\*

**11. References**

Mehmet Ali Turan Kocer

Paper 37, FAO, Rome.

1957; 49: 1507-1521.

England.

Corresponding Author

 \*

Bulent Sen, Feray Sonmez and Ozgur Canpolat

Recently, algae have become significant organisms for biological purification of wastewater since they are able to accumulate plant nutrients, heavy metals, pesticides, organic and inorganic toxic substances and radioactive matters in their cells/bodies with their bioaccumulation abilities. Particularly, biological wastewater treatment systems with micro algae have gained great importance in last 50 years and it is now widely accepted that algal wastewater treatment systems are as effective as conventional treatment systems. Removal rates of particularly high rate algal ponds are almost similar to conventional treatment methods but it is more efficient with lower retention time. With these spesific features algal wastewater treatment systems can be accepted as an significant low-cost alternatives to complex expensive treatment systems particularly for purification of municipal

*University of Firat, Faculty of Fisheries Department of Aquatic Basic Sciences Elazğ, Turkey*

*University of Mersin, Faculty of Fisheries Department of Aquatic Basic Sciences Mersin, Turkey*

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[3] Altenburger, R., Backhaus, T., Boedeker, W., Faust, M.,Scholze, M., Grimme, L.H. Predictability of the toxicity of multiple chemical mixtures to Vibrio fischeri: mixtures composed of similarly acting chemicals. Environ. Toxicol. Chem. 2000; 19, 2341\_/2347. [4] Sigworth, E.A. Control of odor and taste in water supplies. J.Amer. Water Wks. Assn.

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*Mediterranean Fisheries Research Production and Training Institue, Antalya, Turkey*

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[47] Ting, Y.P., Lawson, E. and Prince, I.G. Uptake of cadmium and zinc by alga Chlorella

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**Chapter 15**

© 2013 Raicevic et al., licensee InTech. This is an open access chapter distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

© 2013 Raicevic et al., licensee InTech. This is a paper distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

**Eutrophication: Status, Trends**

Vera Raicevic, Mile Bozic, Zeljka Rudic, Blazo Lalevic,

Dragan Kikovic and Ljubinko Jovanovic

http://dx.doi.org/10.5772/50558

cause fish kills in midsummer [2].

beyond the lake's natural capacity.

**1. Introduction**

Additional information is available at the end of the chapter

**and Restoration Strategies for Palic Lake**

Eutrophication is the general term used by aquatic scientists to describe the suite of symptoms that a lake exhibits in response to fertilization with nutritients [1]. Common symptoms include dense algal blooms causing high turbidity and increasing anoxia in the deeper parts of lakes from the decay of sedimenting plant material. The anoxia can in turn

Aging process is a principal characteristic of each lake. This process is generaly very slow in geological sense, but the aging of shallow lakes (like Palic lake is) is much faster. Eutrophication is not necessarily harmful or bad; it's a Greek word, which means "well nourished" or "good food." However, eutrophication can be artificially accelerated, eventually leading to the suffering of its inhabitants, as the nutrients input increases far

Eutrophication of lakes and reservoirs is a degradation process originating from the introduction of nutrients from agricultural run-off and untreated industrial and urban discharges [3]. Accelerated eutrophication of lakes and reservoirs experienced during last century in most parts of the world represents a serious degradation of water quality, not

Surface waters are not isolated from their environment, neighter from human activities. Therefore, the inorganic and organic matter continuously feed these waters and consequently accelerate the process of natural eutrophication. Waters coming from treatment plants in Subotica town, have been loading this lake for decades, attributing to high nitrogen and phosphorus loadings of Palic Lake together with numerous diffuse pollutants from surrounding villages and agricultural surfaces. Agriculture has been

only in the developing countries [4-6], but also in developed countries [7-10].

**Chapter 15**

## **Eutrophication: Status, Trends and Restoration Strategies for Palic Lake**

Vera Raicevic, Mile Bozic, Zeljka Rudic, Blazo Lalevic, Dragan Kikovic and Ljubinko Jovanovic

Additional information is available at the end of the chapter

http://dx.doi.org/10.5772/50558

## **1. Introduction**

354 Water Treatment

[59] DOE,. Alternative Wastewater Treatment: Advanced Integrated Pond Systems, US Departmant of Energy, Office of Energy Efficiency and Renewable Energy 1993, Technical Information Program Document No: DOE/CH100093-246, Washington. 8p.

> Eutrophication is the general term used by aquatic scientists to describe the suite of symptoms that a lake exhibits in response to fertilization with nutritients [1]. Common symptoms include dense algal blooms causing high turbidity and increasing anoxia in the deeper parts of lakes from the decay of sedimenting plant material. The anoxia can in turn cause fish kills in midsummer [2].