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Рекультивация загрязненных нефтью почв с помощью дождевых червей Dendrobaena Veneta и комплекса микроорганизмов
Поступила 25.11.2018 г.
© Чачина Светлана Борисовна', Бакланова Ольга Николаевна2, Чачина Елизавета Павловна'
1 Федеральное государственное бюджетное образовательное учреждение высшего образования «Омский государственный технический университет», г. Омск, Россия
2 Институт проблем переработки углеводородов Сибирского отделения Российской акадмии наук, г. Омск, Россия
Аннотация. Известно, что дождевые черви ускоряют процесс удаления различных загрязнений из почвы. Дождевые черви изменяют физические и химические свойства почвы, смешивая ее с органическими веществами путем их зарывания в почву, тем самым происходит улучшение аэрации, а загрязняющие вещества переходят в доступные для микроорганизмов формы. Предлагется метод биоремедиации нефтезагрязненных почв с концентрацией нефти до 100 г/кг почвы, в присутствии дождевых черей D. veneta и бактерий bacteria Paenibacillus pabuli, Azotobacter vinelandii, Lactobacillus casei, Clostridium limosum, Cronobacter sakazakii, Rhodotorulla mucilaginosa, Cryptococcus albidus, yeast Saccharomyces, Candida lipolitica, Candida norvegensis, Candida guilliermondii, fungi Aspergillus и Penicillium также Actinomycetales (KOE UFC)=2*1011 per mL). В течение эксперимента, длившегося 5 месяцев, наблюдалось значительное снижение (на 95...97%) углеводородов, зарегистрированное в почве в присуствии дождевых червей и биопрепарата из бактерий.
Ключевые слова. Рекультивация, дождевые черви, микроорганизмы, загрязнение почвы.
The recurvation of oilcontaminated soils with the help of earthworms Dendrobena veneta and the complex of microorganisms
Received on November 25, 2018
© CHachina Svetlana Borisovna', Baklanova Olga Nikolaevna2, CHachina Elizaveta Pavlovna'
' Omsk State Technical University, Omsk, Russia
2 Institute of Hydrocarbons Processing of Siberian Branch of the Russian Academy of Sciences, Omsk, Russia
Abstract. The earthworms are known to enhance the process of contaminants removal from the soil. The earthworms change physical and chemical properties of the soil by mixing it with organic substances through burrowing living habits thus improving the aeration and making contaminants accessible to the microorganisms. The method of oil contaminated soil bioremediation with oil concentrations up to 100 g/kg, in the presence of bacteria Paenibacillus pabuli, Azotobacter vinelandii, Lactobacillus casei, Clostridium limosum, Cronobacter sakazakii, Rhodotorulla mucilaginosa, Cryptococcus albidus, yeast Saccharomyces, Candida lipolitica, Candida norvegensis, Candida guilliermondii, fungi Aspergillus and Penicillium as well as Actinomycetales (KOE UFC)=2*1011 per mL) and D. veneta is proposed. During the experiment lasting for 5 months, a significant lowering (by 95...97%) in hydrocarbons content was registered in the soil with the earthworms and the biopreparation.
Keywords. Reclamation, earthworms, microorganisms, soil pollution.
The earthworms are known to enhance the process of contaminants removal from the soil. The earthworms change physical and chemical properties of the soil by mixing it with organic substances through burrowing living habits thus improving the aeration and making contaminants accessible to the microorganisms [1]. Lifecycle of Dendrobena veneta proceeds at 15 °C and ends in 100-150 days, but at 25 °C the maturing progresses faster, the earthworms produce cocoons earlier and the number of cocoons per
container is higher than at 15 °C. Cocoons incubation period is shorter than at a higher temperature [2].There are data on Dendrobena veneta use in lead removal from the soil with high concentration of lead and cadmium [3]. The information on heavy metals kinetics in the earthworms Dendrobaena veneta was accumulated. Marinussen M.P. et al. made some tests on metals accumulation and removal in laboratory conditions using the soils contaminated with the heavy metals (Cu, Pb, Zn) [4]. The worms were incubated for
28-112 days in the soil contaminated with heavy metals (242 mg/kg of Cu, 109 mg/kg of Pb, 72 mg/kg of Zn). On day 112, an unexpected increase of Cu and Zn concentrations in the worms tissue was noticed [5].
Erlacher et al. studied the influence of various oil concentrations on survivability of Dendrobena hortensis. At concentration 823 mg/kg the mortality of Dendrobena hortensis rose up to 100%. But after amendment with organic substrate, the mortality did not exceed 60% at higher oil concentrations from 1059 to 2241 mg/kg [6]. Hickman Z.A. and Reid noticed the decrease of 3-methylchlorantrene concentration in soil D. veneta cultivation[7].
Therefore, the undertaken analysis of the publications shows that the information on D. veneta application in oil-contaminated soil recultivation is insufficient. The earthworms D. veneta were used in oil-contaminated soil boiremediation at hydrocarbons concentration of no more than 10 g per 1 kg of soil, and it was proved to me an efficient method.
Research results. Total population of D. veneta. The numeration of the curves corresponds to the numbers of the samples in Table. As seen in Figure, after 22 weeks of incubation, control samples 1 and 2 containing 10 adult earthworms D. veneta (sample 1) and 10 adult earthworms D. veneta + microorganisms (biopreparation Baykal EM) (sample 2) showed an increase in the total population of the earthworms with 33 species for sample 1 and 95 species for sample 2. As one can see in Figure, the addition of biopreparation Baykal EM to the soil (sample 2) accelerated the growth of earthworms numbers in comparison with sample without biopreparation Baykal EM. The introduction of crude oil into the soil at a concentration of 20...40 g/kg (samples 3...6) resulted in 80...100% survival of D. veneta and a stable growth of the adult earthworms population both in the presence and in the absence of the microbiological preparation. The earthworms population after 22 weeks of incubation increased to 93 species for sample 3 and to 114 species for the sample with crude oil and biopreparation Baykal EM, i.e., by a factor of 10. As the petroleum content in the soil was increased to 40 g/kg (samples 5 and 6), the earthworm amount in-
crease becomes lower than in samples 3 and 4 and equals 73 species in sample 5 and 40 species in sample 6 after a 22-weeks incubation. An increase in the petroleum content in the soil to 60...100 g/kg (samples 7, 9, 11), led to 100% death of all species in 3 days, which may be caused by chemical burns of the earthworms residing on the surface of contaminated substrate. To increase survivability of earthworms in the soils containing oil at a concentration of 60, 80 and 100 g/kg, soil samples 7, 9, and 11 were supplemented with 1 ml of biopreparation Baikal EM; after that, the oil-contaminated soil samples were held for 4 weeks. As seen in Fig. 1, after a 4-week incubation the survivability of D. veneta rose to 60 %. A further extension of incubation time from 7 to 22 weeks increased the number of earthworms in soil samples. According to Fig. 1 an increase in oil content of the soil decreased the activity of earthworms, and after a 22-week incubation their number in soil samples 8,
10 and 12 containing 60, 80 and 100 g of crude oil and biopreparation Baikal EM, respectively, was 20, 20 and 24 species.
Oil hydrocarbons decomposition. Figure shows the dependence of oil content changes on the incubation time in contaminated soil samples with earthworms D. veneta and biopreparation Baykal EM at the temperature of 15... 17 °C. As Figure shows, in control samples contaminated with oil, the petrochemicals content was changing slightly within the entire incubation period. Addition of biopreparation Baykal EM to the soil samples results in more pronounced hydrocarbons removal. After 22 weeks of incubation, oil content decreased almost twice in the control sample with oil concentration of 20 g/kg (Figure) and biopreparation Baykal EM. Addition of 20 g
011 per 1 kg soil resulted in no significant concentration decrease in the control sample. After 5 months the concentration lowered to 16.73 g/kg. Soil bioremediation proceeds more actively in oil-contaminated samples containing the earthworms D. veneta with oil concentration of 20 g/kg. Hydrocarbons concentration in the soil samples with the earthworms lowered by 97...99% and was equal to 0.95 g/kg. Microbiological preparation amendment resulted in concentration decrease to 9.09 g/kg. Addition of biopreparation Baykal EM with the earthworms D. veneta to the soil samples with 20 g/kg oil amendment resulted in concentration decrease to 0.5 g/kg.
The state of D. Veneta vermipopulations in oilcontaminated soils after 22 weeks of incubation
Sample №№ Survival, % Total population pcs. Total productivity cocoons/container Individual productivity cocoons / species
1. control 100 33±0.98 3.4±0.05 1.5±0.01
2. microbiological preparation 100 95±2.05 15.4±0.12 2.1±0.01
3. soil with oil concentration 20 100 93±2.02 15.3±0.11 1.67±0.01
g/kg 4. soil with oil concentration 20 100 114±2.34 13.4±0.09 1.24±0.02
g/kg and biopreparation
5. soil with oil concentration 40 80 73±1.74 10.7±0.09 1.72±0.02
g/kg 6. soil with oil concentration 40 80 40±0.97 6.8±0.05 0.8±0.01
g/kg and biopreparation
7. soil with oil concentration 60 0 0 0 0
g/kg 8. soil with oil concentration 60 70 20±0.51 3.2±0.02 0.5±0.01
g/kg and biopreparation
9. soil with oil concentration 80 0 0 0 0
g/kg 10. soil with oil concentration 80 60 20±0.51 2.4±0.02 0.3±0.01
g/kg and biopreparation
11. soil with oil concentration 100 0 0 0 0
g/kg 12. soil with oil concentration 100 60 24±0.56 2.5±0.02 0.27±0.01
g/kg and biopreparation
Changes in the concentration of oil hydrocarbons in the soil upon incubation of oil contaminated soil samples in the presence of earthworms D. veneta and biopreparation Baykal-EM in the laboratory experiment. Numeration of the curves corresponds to the numbers of samples in Table: 1 - control sample oil 20g/kg, 2 - control sample oil 20g/kg with Baykal EM-1, 3 - sample with 20 g oil kg-1 and D. veneta, 4 - sample with Baykal EM-1 and 20 g oil kg-1 and D. veneta, 5 - sample with 40 g oil kg-1 and D. veneta, 6 -sample with Baykal EM-1 and 40 g oil kg-1 and D. veneta, 8 - sample with Baykal EM-1 and 60 g oil kg-1 and D. veneta, 10 - sample with Baykal EM-1 and 80 g oil kg-1 and D. veneta, 12 - sample with Baykal EM-1 and 100 g oil kg-1 and D. veneta
The addition of 40.000 g/kg oil decreased the TPH concentration to 35.7 g/kg while the addition of biopreparation Bayka- EM TPH concentration decreased it to 21.5 g/kg after 22 weeks.. In the version with D. veneta, oil concentration lowered to 1.14 g/kg (97% efficiency), and in the sample with the biopreparation it lowered to 2.45 g/kg (93% efficiency).
The addition of 60 g/kg oil decreased the TPH concentration to 47. g/kg after 22 weeks. In the soil sample containing biopreparation Baykal -EM and 60g / kg oil, the TPH concentration decreased to 33.5 g/kg after 22-week incubation, while in the soil sample with D. veneta and biopreparation Baykal-EM, the TPH concentration decreased by 3.3 g/kg(Fig. d). The addition of 80g /kg oil decreased the TPH concentration to 69.8 g/kg. Upon incubation of the soil sample containing 80g/kg oil and Baykal-EM, the TPH concentration decreased to 38.7 g/kg .As for the samples with oil concentration of 80 g/kg and earthworms D. veneta along with biopreparation Baykal-EM, oil content decreased by 99% and equaled 3.6 g/kg. (Fig. c). The addition of 100 g/kg oil decreased the TPH concentration to 85.7 g/kg. Upon incubation of the soil sample containing 100 g/kg oil and Baykal-EM, the TPH concentration decreased to 47.7 g/kg (Fig. e). An increase in the oil content to 100 g/kg hindered the soil sample bioremediation in the presence of earthworms and biopreparation Baykal-EM in soil the TPH concentration decreased by 86 % and was equal to 5.5 g/kg.
Results and discussion. High survivability of the earthworms was registered in our study at oil concentrations up to 50 g/kg (90...97%), probably caused by the soil substrate composition, including nitrogen. Oil concentration more than 50 g/kg results in the death of earthworms D. veneta in 3-7 days. Microbiological preparation introduction improved the earthworms survival in oil-contaminated substrate. Consequently, microbioloigical preparation introduction allows to use D. veneta in recultivation of oil-contaminated oil at higher concentrations of oil, that being impossible so far.
Aurors of this article carried out statistical analysis of the obtained results. Verification of normality of distribution of quantitative traits was performed using the Kolmogorov-Smirnov, Lilliefors and Shapiro-Wilk (W). Aurors of this article are using normality tests (Lilliefors and Shapiro-Wilk tests) the resulting value was less (p < 0.05), therefore, an alternative
hypothesis was formulated, i.e. the characteristic distribution differs from the normal one. At the initial stage of the investigation, the relations between soil contamination and the number of D. veneta were evaluated on all stages with Spearman's correlation analysis. When evaluating the results by the significance with Kruskal-Wallis criterion in several independent groups at soil contamination of 20 g/kg and 40 g/kg with and without the biopreparation, no statistically significant differences were found. The differences were found between the following samples: the control and the one with 20 g/kg oil and biopreparation Baykal-EM (H ( 3, N= 48) =10.97 p =0.011) : the control and the one with 40 g/kg oil H ( 3, N= 48) =9.637 p =0.0219); the control and the one with 40 g/kg and the biopreparation Baykal- EM( H ( 3, N= 48) =11.29 p =0.01). At oil concentration of 60 g/kg-100 g/kg there are differences between the control sample and the one with 100 g/kg oil (Н) H ( 4, N= 60) =15.44 p =0.0001, while in other cases the decrease in the earthworms' amount was registered at concentration increase.
Conclusions The method of oil contaminated soil bioremediation with oil concentrations up to 100 g/kg, in the presence of bacteria Paenibacillus pabuli, Azotobacter vinelandii, Lactobacillus casei, Clostridium limosum, Cronobacter sakazakii, Rhodotorulla mucilaginosa, Cryptococcus albidus, yeast Saccharomyces, Candida lipolitica, Candida norvegensis, Candida guilliermondii, fungi Aspergillus and Penicillium as well as Actinomycetales (KOE UFC)=2*1011 per mL) and D. veneta is proposed. During the experiment lasting for 5 months, a significant lowering (by 95...97%) in hydrocarbons content was registered in the soil with the earthworms and the biopreparation.
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Дополнительная информация
Сведения об авторах:
Чачина Светлана Борисовна, кандидат биологических наук, доцент; Федеральное государственное бюджетное образовательное учреждение высшего образования «Омский государственный технический университет»; Российская Федерация, г.Омск, пр-т Мира, д. 11; e-mail: ksb3@yandex.ru.
Бакланова Ольга Николаевна, кандидат технических наук, старший научный сотрудник; Институт проблем переработки углеводородов Сибирского отделения Российской акадмии наук; Российская Федерация, г.Омск, ул Нефтезаводская, д. 54; e-mail: baklanova@ihcp.ru.
Чачина Светлана Борисовна, студентка; Федеральное государственное бюджетное образовательное учреждение высшего образования «Омский государственный технический университет»; Российская Федерация, г.Омск, пр-т Мира, д. 11; e-mail: ksb3@yandex.ru.
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Для цитирования: Чачина С.Б., Бакланова О.Н., Чачина Е.П. Рекультивация загрязненных нефтью почв с помощью дождевых червей Dendrobaena Veneta и комплекса микроорганизмов // Экология и строительство. 2018. № 4. C. 34-38. doi: 10.24411/2413-8452-2018-10018.
Additional Information
Information about the authors:
CHachina Svetlana Borisovna, candidate of biological sciences, docent; Omsk State Technical University; PR. Mira 11, Omsk, 644011, Russia; e-mail: ksb3@yandex.ru.
Baklanova Olga Nikolaevna, candidate of technical sciences, senior researcher; Institute of Hydrocarbons Processing of Siberian Branch of the Russian Academy of Sciences; 54 Neftezavodskaya Str, 644040 Omsk, Russian Federation; e-mail: baklanova@ihcp.ru.
CHachina Elizaveta Pavlovna, student; Omsk State Technical University; PR. Mira 11, Omsk, 644011, Russia; e-mail: ksb3@yandex.ru.
This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons license, unless i n-dicated otherwise in a credit line to the material. If material is not included in the article's Creati ve Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.
For citations: Chachina S.B., Baklanova O.N., Chachina E.P. The recultivation of oilcontaminated soils with the help of earthworms Dendrobena veneta and the complex of microorganisms // Ekologiya i stroitelstvo. 2018. № 4. P. 34-38. doi: 10.24411/2413-8452-
CHachina Svetlana Borisovna, candidate of biological sciences, docent; Omsk State Technical University; PR. Mira 11, Omsk, 644011, Russia; e-mail: ksb3@yandex.ru.
Baklanova Olga Nikolaevna, candidate of technical sciences, senior researcher; Institute of Hydrocarbons Processing of Siberian Branch of the Russian Academy of Sciences; 54 Neftezavodskaya Str, 644040 Omsk, Russian Federation; e-mail: baklanova@ihcp.ru.
CHachina Elizaveta Pavlovna, student; Omsk State Technical University; PR. Mira 11, Omsk, 644011, Russia; e-mail: ksb3@yandex.ru.
This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons license, unless i n-dicated otherwise in a credit line to the material. If material is not included in the article's Creati ve Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.
For citations: Chachina S.B., Baklanova O.N., Chachina E.P. The recultivation of oilcontaminated soils with the help of earthworms Dendrobena veneta and the complex of microorganisms // Ekologiya i stroitelstvo. 2018. № 4. Р. 34-38. doi: 10.24411/2413-84522018-10018.
