Method for determining toxic effect of waste water on aqueous saline media

FIELD: ecology.

SUBSTANCE: method for determining the toxic effect of waste water on aqueous saline media refers to aquatic toxicology and aims at assessing the toxic effect of an aqueous medium containing waste water. The method consists in determining the unicellular algal culture growth rates in the tested water and involves growing the unicellular algal culture, performing a biotesting procedure consisting in water sampling, introducing a grown algal sub-culture into a reference and a test medium, and counting the algal cells. The test objects are presented by the unicellular microalgae Platymonas viridis Rouch. and Dunaliella salina Teod, on which a long-term (15-day) experiment is carried out. Microalgae Platymonas viridis Rouch are used to determine the toxic effect of waste water on the saline medium.

EFFECT: improving the method.

 

The alleged invention relates to aquatic toxicology and is intended to assess the level of toxicity of the marine environment containing waste water.

Currently, the problem of emergency and planned discharges of different origin in the waters of Ukraine is very important. In this regard, there is the problem of assessing the toxicity of wastewater, both for aquatic ecosystems and for human health, use of water resources. Wastewater settlements, represented by the runoff of house sewers and cattle yards, plums from the garden and horticultural farms, private hospital facilities and enterprises of light and heavy industry, in most cases, the chemical composition is fresh. Once in freshwater, such waste will certainly cause changes in the chemical composition of the water. To assess the toxicity of the environment containing waste water, used methods, based on the determination of the effects of acute (short-term) toxic concentrations of effluent to freshwater aquatic organisms. At the same time, methods for determining toxicity of effluents to aquatic organisms and aquatic organisms living in waters with high salinity is very limited. Therefore, staff treatment facilities and workers SES continue to follow what medicame, designed for freshwater biota, even if the discharge of wastewater from settlements are carried out in the sea or the great salt lake.

Known (see C. E. Dyatlov, A. G. Petrosyan Phaeodactylum tricomutum Boil. (Chrysophyta) as the test object. General provisions // Phycology.- 2001. - So 11, No. 1. -S. 145-154) method of assessing the toxicity of the marine environment, which was the basis of the normative document "Bestaande morco water that Stoney, Yak dadida in the sea. The methodology approved by the Ministry of ecological safety of Ukraine No. 46 from 30.05.1995, (KND, 1995). In a known method of biotesting of sea water is carried out with the use of marine unicellular Alga Phaeodactylum tricomutum. The method is based on determining the growth of the culture of the marine unicellular Alga Phaeodactylum tricomutum in the tested water. The criterion of toxicity is a significant change in the number of algae cells, exhibited in the test environment, compared to control, within 72 hours. The method comprises the cultivation of a culture of the marine unicellular Alga Phaeodactylum tricomutum, the procedure biotesting and processing results. Procedure biotesting consists of water sampling, depositing in the control and in the test environment inoculum cultivated algae, counting the number of cells of the Alga Phaeodactylum tricomutum at 24, 48 and 72 h exposure. The authors proposed a scale of toxicity testirovanie on growth rates of algae. The main result of these works is the determination of the minimum dilution ratio (min Cu) - relationship test water to control. In addition, the authors refer to the work of defining the minimum medienallee effective (lethal) concentrations.

There is a method of determining the toxicity of wastewater in laboratory culture algae Phaeodactylum tricomutum has a number of disadvantages:

is intermittent rapid test and does not allow to evaluate the different phases of culture growth, namely the periods of indifference, the initial disturbance of the functions of the cells, the period of normalization or even stimulate growth. It is chronic (long-term) experiment, the researcher can receive the full information about toxicity test environment;

- does not give complete and accurate information when assessing the risk of discharges of different origin for salinity waters.

The basis for the invention "method for determining the toxicity of wastewater on water salty environment goal by conducting research on the impact of wastewater toxicity to the unicellular algae Platymonas viridis Rouch and Dunaliella salina Teod. in a long term experiment, to provide researchers a more complete and reliable information about the degree of toxicity of different concentrations of wastewater.

p> This objective is achieved in that in the method by determining the growth culture of marine unicellular algae in the test water, including the cultivation of a culture of the marine unicellular algae, the procedure biotesting consisting of water sampling, depositing in the control and in the test environment inoculum cultivated algae, counting the number of algae cells, as test objects have a culture of unicellular marine algae Platymonas viridis Rouch and Dunaliella salina Teod and conduct long-term experiment. Cuts Platymonas viridis Rouch used to assess the toxicity of effluent on the marine environment. Cuts Dunaliella salina Teod. used to assess the severity of discharges of different origin for salinity waters.

The difference of the proposed method against known is that the author proposes to study the toxicity of wastewater on unicellular algae at long-term (15-day) experiment. In addition, the difference of the proposed method from the known, is used as test-objects of culture Platymonas viridis Rouch-to assess the toxicity of the effluent on the marine environment and Dunaliella salina Teod - to assess the risk of discharges of different origin for salinity reservoirs. These types of micro the algae were selected due to the following:

- easy to culture and maintain in laboratory conditions;

- the ability to quickly obtain biomass crops for a short time;

- high sensitivity to the action of toxicants;

- wide use as test objects in ecotoxicology;

the relative stability of P. viridis and D. salina to action of some toxicants, including waste waters of different origin;

- accessibility: the Alga P. viridis is widely distributed in marine waters, a D. salina is a typical representative of the water bodies with a high degree of salinity.

The method is implemented as follows. For biotesting use algological pure culture from the Museum of the cultures of algae, Institute of biology of the southern seas NAS of Ukraine, Sevastopol: Platymonas viridis (Rouch, 1970) - green Alga of the SEM. Chlamydomonadae (Chlorophyta, Volvociphyceae) and green unicellular flagellate halophilic Alga Dunaliella salina (Teod, 1905) from SEM. Chlamydomonadae, (Chlorophyta, Volvociphyceae). For periodic biotesting sea and salinity of water containing waste water, microalgae cultivated under laboratory conditions using a nutrient medium Goldberg (for algae Platymonas viridis and Dunalialla salina), environment Erd-Schreiber (Butcher, 1952) - for cultivation of Platymonas viridis and environment Artery No. 1, No. 2 (1916) and Milko (1962) - for cultivation Dunalialla salina. The cultivation conditions

Culture of algae grown in conical flasks of 500 ml at a temperature of 18 - 25°C and natural light. As the nutrient medium use medium Goldberg. Sea water is taken in a relatively clean area of the sea, filtered through a double filter paper No. 6. Sterilized sea water by triple heating in a water bath to a temperature of 80°C.

Procedure biotesting:

1. The experiment:

Consistently contribute to the marine environment, components of environment Goldberg,

different concentrations of wastewater, as well as an equal number of microalgae culture.

- Select the cell inoculum at the beginning of the experience, then on the 1st, 3rd, 6th, 9th, 12th and 15th day 15-day experiment and calculate the number of cells for each case. The number of cells counted in the entire volume of the chamber Goryaeva, the movement of microalgae fixed with Lugol solution with glycerin.

The experiments performed in triplicate. For each vessel, the number

cells are also determined in triplicate.

2. Statistical processing of the experimental results. The results of the experiments are treated statistically using t - student test when comparing parameter number at a significance level of p<0.05 (Lakin, 1973).

3. Determining the presence or absence of toxic effect of R is slichnih concentrations of wastewater to the culture of algae.

Indicators biotesting

The main index, which evaluates the level of added toxicity in the environment of wastewater, is the number of algae cells. According to the results of the experiments are table cell population in all variants of the experiment in absolute units, as well as values of the number of algal cells, expressed in relative units (% relative to control). Additional indicators of the toxicity of the environment on microalgae, can serve as the percentage of inhibition of cell population, the ratio of the relative growth of crops, the average rate of growth, the percentage of reduction in the average growth rate, the growth rate and the percentage reduction in growth of the culture.

On the basis of the obtained experimental results conclude minimum ratio of dilution of wastewater, sea water or water with high salinity, that is, establish such number of natural water, which will not be observed toxic effect of impurities. To detect toxicity at various concentrations of wastewater, the author also offers guided "Scale toxicity test environment for the growth of algae Phaeodactylum tricomutum Bohl" (woodpeckers, Petrosyan, 2001), which is the percentage of the number of cells in solution determines the degree of toxicity and the nature of istica environment, as well as the rate of dilution. Some researchers also determine the maximum allowable (inactive) concentration and lethal concentration (LC50, LC100), which are mainly used in the calibration of crops in relation to the standard action of a toxicant.

Examples of implementation of the method.

Example 1. Evaluated the toxicity level of urban domestic wastewater (HBSW) sea cuts Platymonas viridis Rouch. The results of the experiment are presented in table.1, and in Fig. 1 - Fig. 6.

Table 1

Change in the number of cells Platymonas viridis Rouch. when exposed to various concentrations of domestic wastewater

Note. Numerator - the number of cells (X 10) per ml in the denominator is the same in relation to population control, taken as 100%. The asterisk indicates the value of the number of cells, significantly different from those values in the control cultures (p≤0,05).

On the basis of the obtained results we can conclude that in the long-term experiment HBSW in concentrations of 1, 10 and 100 ml/l did not cause toxicity in the marine flagellate Platymonas viridis Rouch. A significant reduction of more than 50% compared with the control population of cells 9 - 12 day under the influence of wastewater can explain the decrease is receiving an average growth rate of the culture (see Fig. 3), and the percentage reduction of the average rate of growth of microalgae above in step 1, and 100 ml/l of wastewater (see Fig.4). In absolute values, the number of cells of P. viridis were increased in all variants during the whole experiment. However, from table. 1, figs. 1 and Fig. 2 shows that this ratio was higher in the control than in the experiments. It should be noted that the percentage of inhibition abundance was higher at the lowest concentration, and the coefficient of relative growth, respectively, was lower in this case, and also under the influence of 100 ml/l of wastewater. Daily gains of culture of P. viridis was higher in the case of impact HBSW at a concentration of 1 ml/l (see Fig. 5). The decrease in growth was increased with increasing concentration of wastewater (see Fig. 6), which, of course, is consistent with the data table. 1 and shows that the greatest increase in the number of cells occurs when the increase in the content of nutrients present in the wastewater.

Example 1 shows that during the exponential growth period, the culture of P. viridis wastewater urban collector at concentrations of 1, 10 and 100 ml/l did not cause irreversible disturbances in the growth in the number of cells studied culture, though, almost during the whole experiment investigated the rate was lower than the control, which probably indicates excessive nutrient and that is, or toxic components of the effluent.

Example 2. Evaluated the toxicity level of urban domestic wastewater (HBSW) on halophilic cuts Dunaliella salina Teod. The results of the experiment are presented in table.2, and in Fig.7 - Fig. 12.

Table 2

Change in the number of cells of Dunaliella salina Teod. when exposed to various concentrations of domestic wastewater

Note. Numerator - the number of cells (X 104in ml, the denominator is the same in relation to population control, taken as 100 %. The asterisk indicates the value of the number of cells, significantly different from those values in the control cultures (p≤0,05).

Based on the data of experiment to study the effect of sewage on cuts Dunaliella salina Teod (see tab. 2) we can conclude on the safety testing of wastewater in a concentration of 1, 10 and 100 ml/l, the Percentage of inhibition of the number of cells of D. salina was increased with increasing concentration of HBSW (Fig. 7). With 6-day coefficient of relative growth was greatest in step 1, and 10 ml/l of wastewater, while in the control, and, especially, the influence of the wastewater at a concentration of 100 ml/l this indicator was lower. The same results have shown the calculation of the average growth rate of the culture: when 1 and 10 ml/l the growth rate was maximum, and at the highest concentration the AI minimum (Fig. 9). Compared with the control, the percentage of reduction in the average growth rate in the number of cells of D. salina was lowest when exposed to 10 ml/l of wastewater (Fig. 10). Daily growth rate of the culture studied microalgae was higher, the lower the concentration tested HBSW (Fig. 11) and, accordingly, the percentage reduction in growth culture was the lower, the higher the dose of a toxicant (Fig. 12).

Thus, HBSW in concentrations of 1 and 10 ml/l caused on average a greater stimulation of growth in the number of cells of D. salina than 100 ml/L. Stimulation was also observed during the whole experiment, and the maximum, compared with the control, the increase in the number of cells accounted for 3 - 12 hours. At the same time, the study of the effect of wastewater at a concentration of 100 ml/l showed that the same period in culture took place from 3 to 9 days. In all cases the increase in the number of cells in absolute units occurred gradually during the whole experiment (table. 2).

From these examples it follows that the dilution of wastewater urban collector of sea water in the ratio 1:1000 (1 ml/l), 1:100 (10ml/l) and 1:10 (100 ml/l) does not cause irreversible changes in the number of cells of marine algae Platymonas viridis Rouch. and halophilic microalgae Dunaliella salina Teod.

The advantages of the proposed method in comparison with the known are as follows:

the first Avenue is to bring out a risk assessment of discharges of different origin for salinity ponds

- provides a complete and accurate assessment of the degree of toxicity of the investigated medium, because it allows one to evaluate the different phases of culture growth: periods of indifference and primary dysfunction of cells, the period of normalization or even stimulate growth, which is caused either by the creation of an environment favourable conditions, or adaptive abilities of microalgae in response to exposure to a toxicant;

thanks to making long-term experimentai you can identify the time of the most negative manifestations of the dirt.

1. The method of determining the toxicity of wastewater on water salty environment by determining the growth of the culture of the marine unicellular Alga Platymonas viridis Rouch or Dunaliella salina Teod, including the cultivation of algae in the control and in the test environment, the calculation of the number of algae cells, wherein the cultivation is carried out in 15 days, and the number of cells counted at 1, 3, 6, 12 and 15 days of cultivation and population change by 50% or more in the experiment compared to the control is judged on the modifying effect of sewage.

2. The method according to p. 1, characterized in that the cuts Platymonas viridis Rouch used to assess the toxicity of effluent on the marine environment.

3. The method according to p. 1, characterized in that microbodies the ü Dunaliella salina Teod used to assess the toxicity of runoff on salinity reservoirs.



 

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24 cl, 2 dwg, 2 tbl, 1 ex

FIELD: process engineering.

SUBSTANCE: invention relates to biological treatment of domestic sewage effluents. Effluents are fed from collector 1 into anaerobic digester 2. Contaminated biogas is fed from methane fermentation chamber 6 into inlet branch pipe 38 and is forced by pump 11 in discharge branch pipe 39. Contaminants are fed into appropriate collector 45. Processed biogas is fed into chlorella generator 12. Pure methane is collected in collector 12. Chlorella and thiobacteria are forced via branch pipe 16 into dynamic disintegrator 15. Mix of heavy and standard water is fed via hydraulic gate 19 into rectifier 20 to produce heavy and standard water. Heavy water is directed from heat exchanger 28 into circulation circuit 31 of reactor 32. Portion of the mix of standard and tritiated water is directed into extra rectifier 35. Tritiated water is separated from standard water and directed into collector.

EFFECT: higher efficiency of treatment.

2 dwg

FIELD: municipal economy; industrial enterprises; sewage purification.

SUBSTANCE: the invention is pertaining to the methods of biological purification of sewage in bioreactors and may be used in the water drain systems of municipal economies and the industrial enterprises at purification of sewage. The sewage is treated in a bioreactor with membranous separation of water and an active sludge. Conduct a periodic regeneration of the separator. The membrane contains oxidation catalysts in the form of a combination of metals with variable valence, for example - manganese or cobalt oxides. The technical effect is intensification of processes of a biological purification of sewage, an increase of duration of a filtration service life of the membranous separator and reliability of its operation.

EFFECT: the invention ensures intensification of processes of the biological purification of sewage, increased duration of a filtration service life of the membranous separator and reliability of its operation.

1 tbl, 2 ex

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