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Soil balls containing the so-called effective microorganisms (EM) have been applied to improve water quality of small ponds, lakes, and streams worldwide. However, neither the physical conditions facilitating their proper application nor the diversity of microbial community in such soil balls have been investigated. In this study, the application of 0.75 % of hardener to the soil balls exerted almost neutral pH (pH 7.3) which caused up to a fourfold increased hardness of the soil ball. Moreover, the 0.75 % of hardener in the soil ball also improved the water quality due to a significant reduction in dissolved oxygen, total phosphorus, and total nitrogen contents. Metagenomic analysis of the microbial community in the soil ball with 0.75 % hardener was compared with control (traditional soil ball) through next-generation sequencing. The traditional soil ball microbial community comprised 96.1 % bacteria, 2.7 % eukaryota, and 1 % archaea, whereas the soil ball with 0.75 % hardener comprised 71.4 % bacteria, 27.9 % eukaryota, and 0.2 % viruses. Additionally, metagenomic profiles for both traditional and improved soil balls revealed that the various xenobiotic biodegradation, such as those for caprolactam, atrazine, xylene, toluene, styrene, bisphenol, and chlorocyclohexane might be responsible for organic waste cleanup.
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An improved effective microorganism (EM) soil ball-making
method for water quality restoration
Gun-Seok Park
&Abdur Rahim Khan
&Yunyoung Kwak
&Sung-Jun Hong
ByungKwon Jung
&Ihsan Ullah
&Jong-Guk Kim
&Jae-Ho Shin
Received: 9 June 2015 /Accepted: 15 October 2015 /Published online: 24 October 2015
#Springer-Verlag Berlin Heidelberg 2015
Abstract Soil balls containing the so-called effective micro-
organisms (EM) have been applied to improve water quality
of small ponds, lakes, and streams worldwide. However, nei-
ther the physical conditions facilitating their proper applica-
tion nor the diversity of microbial community in such soil
balls have been investigated. In this study, the application of
0.75 % of hardener to the soil balls exerted almost neutral pH
(pH 7.3) which caused up to a fourfold increased hardness of
the soil ball. Moreover, the 0.75 % of hardener in the soil ball
also improved the water quality due to a significant reduction
in dissolved oxygen, total phosphorus, and total nitrogen con-
tents. Metagenomic analysis of the microbial community in
the soil ball with 0.75 % hardener was compared with control
(traditional soil ball) through next-generation sequencing. The
traditional soil ball microbial community comprised 96.1 %
bacteria, 2.7 % eukaryota, and 1 % archaea, whereas the soil
ball with 0.75 % hardener comprised 71.4 % bacteria, 27.9 %
eukaryota, and 0.2 % viruses. Additionally, metagenomic pro-
files for both traditional and improved soil balls revealed that
the various xenobiotic biodegradation, such as those for cap-
rolactam, atrazine, xylene, toluene, styrene, bisphenol, and
chlorocyclohexane might be responsible for organic waste
Keywords Biodegradation .Effective microorganisms .
Metagenome .Microbial community .Soil ball .Water quali ty
Microbial biotechnology is the most recent approach to waste-
water treatment and is essential for protecting human health
and the environment (Mielczarek et al. 2013). In order to
guarantee the optimal operation using the effective microor-
ganism technology, it is very important to understand the
structure, function, and microbial community dynamics in-
volved in these approaches (Zakaria et al. 2010). Soil balls
have been used as carrier to immobilize effective microorgan-
isms for the remediation of contaminated water environment
(Ekpeghere et al. 2012). The concept of Beffective
microorganisms^(EM) was introduced by Dr. Teruo Higa,
and since then, EMs became an important part of natural
farming (Higa 1998). Those microbes are selected based on
their functions as fixation of atmospheric nitrogen, decompo-
sition of organic wastes and residues, suppression of soil-
borne pathogens, recycling and increased availability of plant
nutrients, solubilization of insoluble nutrient sources, and deg-
radation of toxicants including pesticides (Higa and Parr
1994). EM is co-cultures of naturally occurring beneficial mi-
croorganisms, and it is widely applied as inoculants to soil,
water and plants for the improvement of soil and water quality,
as well as for plant growth and crop yield (Grover et al. 2011;
Javaid 2010). Soil balls with EM can be a good alternative for
stream water treatment in eco-friendly environments, but there
have been no report on their physicochemical properties and
Responsible editor: Robert Duran
Electronic supplementary material The online version of this article
(doi:10.1007/s11356-015-5617-x) contains supplementary material,
which is available to authorized users.
*Jae-Ho Shin
School of Applied Bioscience, Kyungpook National University,
Daegu 702-701, Republic of Korea
Institute of Biotechnology and Genetic Engineering, The University
of Agriculture, Peshawar, Pakistan
School of Life Sciences and Biotechnology, Kyungpook National
University, Daegu 702-701, Republic of Korea
Environ Sci Pollut Res (2016) 23:11001107
DOI 10.1007/s11356-015-5617-x
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
... A study by Park et al. [31] also sought to improve the quality of water of streams by studying the physical conditions and the diversity of the microorganisms in the water. Park et al. [31] made use of soil balls that contained EM and found that the quality of water was improved by hardener soil balls as it significantly reduced the dissolved oxygen, total nitrogen, and total phosphorus components. ...
... A study by Park et al. [31] also sought to improve the quality of water of streams by studying the physical conditions and the diversity of the microorganisms in the water. Park et al. [31] made use of soil balls that contained EM and found that the quality of water was improved by hardener soil balls as it significantly reduced the dissolved oxygen, total nitrogen, and total phosphorus components. The composition of the hardener soil ball and traditional soil ball microbial community was found to differ with the hardener soil balls comprising 0.2% viruses, 27.9% eukaryote, and 71.4% bacteria while the traditional soil ball consisted of 1% archaea, 2.7% eukaryote, and 96.1% bacteria. ...
... An assessment of pathways that support the xenobiotic degradation was also performed. An examination at the metagenomic profiles for both the hardened soil and the Contaminated harbor sediments Loess balls containing EM Acetic acid, propionic acid, valeric acid [14] Artificial lake Solution COD, TN, TP, TSS, turbidity [47] Synthetic polluted water Solution NH 3 -N, TP, COD [19] Stream water Soil balls containing EM TP, TN, xenobiotic [31] Artificial river water Mudballs containing EM COD, TSS [23] Safwat and Matta Journal of Engineering and Applied Science (2021) 68:48 Page 4 of 12 traditional soils showed that a certain percentage of the organic waste cleanup was due to the xenobiotic biodegradation [31]. Mudballs containing EM were also applied by a study by Nugroho et al. [23] to examine the impact that temperature had on COD and TSS removal. ...
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Nowadays, beneficial microorganisms are getting wider applicability. One example is referred to as Effective Microorganisms (EM) having its composition kept a secret. EM is a product in liquid form, which consists of a variety of not only effective and beneficial microorganisms but also nonpathogenic ones, with admirable coexistence between aerobic and anaerobic types of microorganisms. The aim of this narrative review is to provide a summary of the different uses and applications of EM, their applications, their benefits, and the expected results when using them in different applications. This is the first review to focus on the uses of EM in environmental engineering systems and processes such as wastewater treatment processes. Originally, EM was manufactured to be utilized in organic farming, but at the moment, this substance is getting wider applications such as in medicine, environment, livestock sector, forestry, and agriculture. When it comes to the protection of the environment, EM helps in waste deodorization, eutrophication control, and wastewater. Investigation on EM use in water quality restoration, wastewater treatment, the treatment of sludge, and composting has been undertaken by researchers. This review provides an overview of the current situation of environmental application of EM in various fields including water quality, wastewater treatment, sludge treatment, and composting.
... En la actualidad, la eutrofización ocasiona graves problemas de deterioro en los ecosistemas acuáticos, donde el exceso de nutrientes como el fósforo y el nitrógeno provenientes de la agricultura, la ganadería y los efluentes de las ciudades propician la generación de floraciones de cianobacterias tóxicas, lo cual constituye un serio problema de salud ambiental y humana que no está siendo abordado (Bonilla, et al., 2015;Chalar, 2006;Goyenola, et al., 2021;Huisman, et al., 2018;Kruk, et al., 2020). En este contexto, se ha reportado el empleo de EM para la remediación de aguas residuales y ha demostrado ser efectivo en la remoción de sólidos en suspensión, disminuyendo la demanda biológica (DBO) y química de oxígeno (DQO) (Okuda y Higa, 1999) (Park, et al., 2016). Sin embargo, son pocos los estudios existentes sobre este tema y la información se encuentra dispersa y poco sistematizada. ...
... Si bien existen numerosos reportes del empleo de los consorcios microbianos denominados microorganismos efectivos como promotores del crecimiento y la salud de los cultivos, compostaje de restos vegetales e incluso descontaminación de suelos radiactivos (Cremeneac y Boclaci, 2018;de Araujo Ávila, et al., 2021;Domenico, 2019;Nikitin, et al., 2018;Pereira, et al., 2019;Pires, et al., 2021), la información acerca de su uso para el tratamiento de efluentes domésticos es escasa. (Park, et al., 2016). En este sentido, se puede atribuir la ausencia de efecto observada en algunos de los pozos sépticos ensayados en este trabajo a la presencia de roturas, fugas o sistemas denominados "robadores" (muy comunes en Uruguay) que sirven a modo de sistema de desagüe cuando el pozo se encuentra muy lleno. ...
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La contaminación fecal es uno de los principales problemas de calidad del agua dulce a nivel mundial. Una de las soluciones propuestas para este problema es el tratamiento de los efluentes a nivel doméstico o local con preparados biológicos tales como los microorganismos efectivos nativos (MEN). En este trabajo se evaluó la eficacia de un preparado de MEN producido en Uruguay para disminuir la contaminación fecal en efluentes domésticos. Para ello se realizaron experimentos a distintas escalas: desde in vitro hasta en pozos sépticos de viviendas y un tramo del arroyo Malvín (in situ), combinando preparado líquido con sólido y empleando a los coliformes fecales (CF) y la materia orgánica particulada como indicadores del rendimiento. En el laboratorio y en los pozos sépticos, los CF disminuyeron significativamente luego de 8 semanas de aplicación. En el arroyo Malvín se observó una disminución de los CF y una recuperación visual del curso de agua, con un aumento de la transparencia y el oxígeno disuelto. Los resultados de este trabajo sugieren que la aplicación de MEN en aguas residuales domiciliarias constituye una aproximación promisoria para reducir la contaminación fecal de los efluentes domésticos y enfatizan la necesidad de cambiar los hábitos de construcción de los pozos sépticos.
... Microbiological biopreparations are the leader in many branches of industry, i.e. the food industry, health care products, cosmetics, purification processes, agriculture, processes of the revitalization of contaminated soils and waters, elimination of odors and pathogenic microorganisms (hazardous to the health of end consumers of humans, animals, and plants) (Zakaria et al., 2010;Park et al., 2016;Sitarek et al., 2017;Dondajewska et al., 2019;Mazur, 2020;Sharip et al., 2020). The process of their production and the methods used are the result of vigorous research and achievements in the scientific discipline of Biotechnology (its various departments). ...
... Among the methods described, regardless of the others, applications of microbiological biopreparations is main method (Mazur, 2020). Many scientific publications present the results of revitalization using this type of technology (Zakaria et al., 2010;Park et al., 2016;Sitarek et al., 2017;Dondajewska et al., 2019;Mazur, 2020;Sharip et al., 2020). Additional treatment-enhancing technologies are mainly based on aeration processes (Carr and Martin, 1978;Dixit et al., 2007;Dong et al., 2012;Chmielowski et al., 2019;Anber et al., 2020), or controlled planting of ecotones in zones of intensive nutrient supply to water (Vought et al., 1994;Fennessy and Cronk, 1997;Holland, 2012). ...
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Aim of the study: The work aims to assess the possibility of the application of selected types of biological beds to support the revitalization processes of strongly degraded water reservoirs. Material and methods: The authors reviewed the literature on biological methods used in the treatment processes of various types of wastewater. Certain types of beds have been selected that show tolerance to temperature changes and significant changes in organic pollutant loads. The self-purification potential of water and the role of natural methods in the revitalization of water reservoirs were characterized. The characteristics of biological methods based on MBBR moving and fixed beds are presented. Results and conclusions: The possibility of application of selected types of MBBR moving and fixed beds in supporting the treatment of highly contaminated surface waters were assessed. Biotechnological methods based on liquid and solid biopreparations normally used in water revitalization were discussed. It has been shown that when biotechnological methods are not able to operate efficiently, it is very beneficial to start additional biological processes to improve the efficiency of the revitalization process.
... Studies have shown that EM can positively influence decomposing organic matter in the environment by facilitating nitrification, denitrification, ammonification, and other processes, improving water and soil quality and maintaining microbial ecological equilibrium (Safwat and Matta, 2021). As a probiotic, it has been widely applied in aquaculture (Abdel-Aziz et al., 2021), organic agriculture (Mwegoha, 2012), wastewater treatment (Park et al., 2016), and other fields and has been demonstrated to have no adverse effects on fish, humans, and environmental conditions (Abdel-Aziz et al., 2020). At present, EM has been adopted by over 100 countries worldwide, not on an experimentation basis but for commercial use as well as environmental management (Olle and Williams, 2013). ...
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Integrated rice and aquaculture farming is a high-intensity agricultural model that efficiently utilizes soil and water resources. However, soil ecological issues caused by overexploitation of this model may jeopardize rice production and food security. Given the requirements of aquatic animal breeding circumstances, the application of biological agents has become the preferred choice for improving the environment in this model. In this study, the rice–crayfish cocropping (RC) model was used as the research object, and the widely used bio-fertilizer Effective Microorganisms (EM) was employed as a biological amendment. This study seeks to clarify the improvement effect of EM on the soil and water environment of the RC model as well as to explore its microbiological mechanism. The findings demonstrated that EM significantly improved the chemical properties of the model, for example, the dissolved oxygen in water increased by about 44%, and the redox potential in soil increased by about 150% over the entire rice growing period. Second, the use of EM reduced the soil microbial diversity while reconstructing a more compact and stable microbial cooccurrence network. The reconstructed community selectively enriched the functions related to photosynthesis and nitrogen transformation to cope with the environmental stresses of low dissolved oxygen or high nitrogen. Subsequently, 30 dominant-ASVs (ASV, Amplicon Sequence Variant) with high abundance, strong functionality, positive correlation with environmental factors, and good representation of the community were identified using correlation analysis, abundance comparison, and model prediction. This study not only verified the application effect of EM in a complex agricultural intensive system but also proved that the community induced by EM has a positive effect on ecosystem multifunctionality and identified the key dominant species. This provides strong evidence for us to understand the microbiological mechanisms of exogenous biological agents in environmental improvement and provides a reference for screening and optimizing functional microbial species in agricultural ecosystems.
... The biotechnology named Effective Microorganisms (EM) was first proposed by Dr. Teruo Higa in Japan in 1982, who mixed beneficial microorganisms [19]. EM bacteria has been widely applied in wastewater treatment [20], decomposing pollutants in the environment by facilitating nitrification, denitrification, ammonification, and other processes, then improving water and soil quality without damages to the environment. A field experiment showed that applying EM bacteria to constructed wetlands could enhance the removal rate and treatment time of nitrogen and phosphorus [21]. ...
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Eco-ditch systems have increasingly been designed and applied as a strategy to decrease the risks of water eutrophication and contamination pollution for sustainable agriculture. The main goal of this study was to evaluate the water quality of eco-ditch substrates amended with zeolite and Effective Microorganisms (EM), such as pH, dissolved oxygen concentration (DO), ammonium nitrogen concentration (NH4 +-N), and nitrate nitrogen concentration (NO3-N). Laboratory experiments were conducted with four single substrates (soil, none substrates, natural zeolite, and zeolite loaded with EM bacteria) and two mixed substrates (soil and varying proportions of the additives, 0, 5 and 15%, m/m). Results showed that the concentration of NH4 +-N was decreased with the increasing rates of additives, and zeolite loaded with EM bacteria had the highest nitrogen removal rate (97.90%) under static experimental condition. The application rate of 15% zeolite loaded with EM bacteria on the eco-ditch exerted a better effect on NH4 +-N and NO3 -N removal without pH reduction, decreased by 87.19% for NH4 +-N and 30.33% for NO3 -N, respectively. Path analysis showed that zeolite addition had a rapid effect (path coefficient = -0.972) on free NH4 +-N ions adsorption in early 1–3 days, then EM loaded at zeolite further decreased NH4 +-N (path coefficient = -0.693) and NO3 -N (path coefficient = -0.334) via bacterial metabolism. Based on the results, the applications of natural zeolite and Effective Microorganisms (EM) at an appropriate rate (15%, m/m) can significantly improve water quality of paddy drainage via exerting effects on nitrogen removal.
... Tương tự đối với nitơ trong nguồn nước, kết quả khảo sát đối với NO 3 -, NO 2 -cũng đạt hiệu quả khá tốt. Cụ thể, nồng độ của chúng giảm từ 0,022±0,015 mg/L xuống 0,006±0,004 mg/L (NO 2 -) và từ 9,60±2,31 mg/L còn 0,44±0,27 mg/L (NO 3 - Có thể thấy, việc áp dụng công nghệ sản phẩm sinh học có tính hiệu quả và được xem như là công cụ hữu dụng phục hồi các hồ bị ô nhiễm [16,17]. Công nghệ sử dụng sản phẩm Bakture làm chất xúc tác, kích thích sự phát triển của vi sinh vật hiếu khí và yếm khí hoạt động, qua đó phân giải các chất độc hại trong môi trường nước. ...
The paper presents the results of the application of Bakture (Back to Nature) aiming to treat polluted water resources in lakes and canals of Thu Duc district, Ho Chi Minh City. The studied objectives were chosen related to heavily polluted surface water in canals and ponds systems. The study selected four polluted surface waters that are affected by different wastewater types (industrial, domestic and husbandry). The experiments were conducted in four glass tanks with each volume of 100L, and size L×W×H is equal to 0.6m × 0.4m × 0.5m. Aiming to examine treated efficiency, this research carried out sampling, analyzing water quality parameters such as pH, TSS, COD, BOD5, N-NH4+, N-NO3-, N-NO2-, P-PO43-, and microorganisms levels. The water samples were taken after 1, 2 and 4 weeks during the experiment. The application of Bakture product after four experiment weeks has obtained some positive findings, such as the treatment efficiencies for BOD5, COD, TN, and TP were 78.1±7%, 95.0±11%, 91.2±3%, and TP 54.6±6%, respectively. In addition, a significant increase in microorganisms levels lead to organic matter degradation, maintaining the habitat for the aquatic system. Results provide the environmental protection solutions towards ecologically sustainable and environmental friendly.
... The Afdera and rock salts were obtained from the Danakil desert, Ethiopia (14.24°N, 40.29°E; Gibtan et al., 2017), and the soil samples were obtained from Uiseong horticultural fields, Korea (36.11°N, 128.19°E;Park et al., 2016). Tidal mudflat samples were obtained from the Taean coast, Korea (36.43°N, 126.16°E). ...
Excavating the molecular details of many diverse enzymes from metagenomes remains challenging in agriculture, food, health, and environmental fields. We present a versatile method that accelerates metabolic enzyme discovery for highly selective gene capture in metagenomes using next-generation sequencing. Culture-independent enzyme mining of environmental DNA is based on a set of short identifying degenerate sequences specific for a wide range of enzyme superfamilies, followed by multiplexed DNA barcode sequencing. A strategy of ‘focused identification of next-generation sequencing-based definitive enzyme research’ (FINDER) enabled us to generate targeted enzyme datasets from metagenomes, resulting in minimal hands-on obtention of high-throughput biological diversity and potential function profiles, without being time-consuming. This method also provided a targeted inventory of predicted proteins and molecular features of metabolic activities from several metagenomic samples. We suggest that the efficiency and sensitivity of this method will accelerate the decryption of microbial diversity and the signature of proteins and their metabolism from environmental samples. This article is protected by copyright. All rights reserved.
... The accuracy measures of the model were tested using the RMSE, R-squared (R 2 ), and adjusted R-squared (R 2 adj ). Additional details of the RMSE are provided in equation (9) in Section (V). R 2 is a factual proportion of how closely the data are fitted to the regression line, and is sometimes called the coefficient of determination. ...
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Monitoring water quality is an important challenge in both developed and developing countries. Remote sensing data can form a highly frequent dataset with acceptable spatial coverage that can be used to remotely monitor water quality. This paper presents a novel automated model for remotely monitoring water quality to address the problem of insufficient samples and save the time and cost of sample collection. The proposed model estimates both optical and non-optical water quality parameters via Sentinel-2A data. A bio-inspired hybrid model of a Binary Whale Optimization Algorithm (BWOA) and Artificial Neural Network (ANN) (BWOA-ANN) is applied to determine the relationship between extracted reflectance values from Sentinel-2A images and analyzed samples. The novelty of this model is to solve two main problems of remote water quality monitoring: poor applicability and low non-optical parameter estimation accuracy. For the first problem, a proposed fully automated model with band selection using the BWOA to automatically select the optimal features (Sentinel-2A bands) that are suitable for each water quality parameter. The second problem is addressed by automatically detecting the relationship between non-optical parameters, such as the total phosphorus, and optical parameters, such as chlorophyll-a. Three datasets with different locations, seasons, and parameters were selected to test the proposed BWOA-ANN. The experimental results demonstrated good regression with a mean ${R}^{2}$ value of 0.916 for optical parameters and 0.890 for non-optical parameters. The proposed model was found to outperform the ANN with an ${R}^{2}$ value higher by 40% and 52% for the optical and non-optical parameters, respectively.
... Other beneficial effects of EM technology reported in literature were not limited to plant growth but also to composting process, waste treatment (water and solid), and phenol degradation [38][39][40][41][42]. ...
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The use of ecofriendly strategies, such as the use of Plant Growth Promoting Bacteria, to improve the yield and quality of crops has become necessary to satisfy the growing demand of food and to avoid the use of chemical fertilizers and pesticides. In this study, we report the effects of an innovative microbial inoculation technique, namely Effective Microorganisms (EM), compared with traditional approaches, on productivity and nutritional aspect of four tomato varieties: Brandywine, Corbarino Giallo, S. Marzano Cirio 3, S. Marzano Antico. Results showed an increase of plant productivity as well as an enhanced antioxidant activity mainly in San Marzano Antico and Brandywine varieties treated with EM technology. Moreover, the polyphenol and carotenoid contents also changed, in response to the plant treatments. In conclusion, the application of EM® technology in agriculture could represent a very promising strategy in agricultural sustainability.
This study investigated the effects of effective microorganism (EM) in reducing the cyanobacterial density and improving the water quality and phytoplankton diversity. Four thousand litre of EM was sprayed over an area of 0.7 ha in a semi-enclosed area of the Putrajaya Lake. Sampling and analysis of water quality and phytoplankton were carried out at three locations over a period of 50 days. The results of the study showed that EM treatment temporarily reduced the cyanobacterial density from 3909.5 to 2200.9 cells/ml after 18 days, but the density began to increase again soon after. Similarly, mean phytoplankton density showed similar pattern to the cyanobacterial trend of decreasing density after application but started to increase 3 weeks afterwards. Cyanobacteria was the dominant phytoplankton group before and during EM treatment. Cyanobacterial dominance decreased 55.2% after EM application resulting in more balanced phytoplankton assemblages 18 days after treatment before regaining its dominance 3 week after EM application. Nostoc sp. was the main cyanobacterial species before-, during and after EM treatment. Only pH significantly differed after the EM treatment; while, other parameters such as dissolved oxygen, chlorophyll-a, transparency showed no significant changes before and after EM application. This study illustrated that EM application did not provide sustained microalgae reduction nor water quality improvement throughout the experiment.
Conference Paper
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Water quality has received considerable attention in allocation processes for maximizing the satisfaction of various sectors. However, pollutant impurities that impede adequate supply of water have a detrimental effect on the quality and harmful for living organisms including aquatic life. For the reduction of water pollution level, various chemical and biological treatments are available but the emergence of an amazing technology of a multiculture of anaerobic and aerobic beneficial microorganisms is presently gaining popularity due to its environmentally friendly nature. This effective microorganism (EM) technology uses naturally occurring microorganisms which are able to purify and revive nature. Applications of EM using the formula known as effective microorganism activated solution (EMAS) have been experimented in several rivers in Malaysia depending on the scale, location, physical and geographical conditions with the principal objective of enhancing and improving the water quality. One of the significant contributions of EM based rehabilitation of polluted and degraded water bodies is to restore aquatic habitats and ecosystems. Existing results of projects via EM technology in solving water quality related problems, and the nationwide campaigns in Malaysia are duly presented. The role of EM-based water restoration approach for sustainability of water resources and prospects of modeling are also discussed. Results clearly demonstrated the effectiveness of this technique for restoration of water quality of degraded/polluted river basin. Valuable lines for further research and acceptance of EM technology for the future are thus suggested as it is believed to be the key to sustained environmental improvement and offers a real opportunity for eco-innovation.
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Parthenium weed (Parthenium hysterophorus L.) is rapidly spreading in many countries around the world and has become a serious threat to nature as well as managed ecosystems. The present pot experiment was conducted in 2005 -2006 to study the effect of parthenium green manure (1, 2, 3 and 4% on fresh weight bases) and EM (effective microorganisms), a biofertilizer, on the growth and yield of wheat (Triticum aestivum L.). EM application was carried out by applying 1 L of 0.2% dilution of the commercial stock EM solution per pot at fortnight intervals throughout the experimental period. Plants were harvested at flowering and maturity stages. Shoot length and dry biomass were gradually increased by increasing the quantity of green manure. There was 272% increase in shoot dry biomass over control at maturity due to 4% green manure application as compared to 137% increase due to recommended dose of nitrogen-phosphorus-potassium (NPK) fertilizer. Highest root biomass was recorded in 3% green manure amended treatment. Spike length, number of grains per spike and grains yield was also gradually increased by increasing the quantity of green manure. There was 43 to 253% increase in grain yield over control due to various green manure treatments as compared to 96% increase due to NPK fertilizers over control. EM application in un-amended control, NPK fertilizers as well as in various green manure treatments either exhibited insignificant effect or significantly reduced various studied parameters. The present study concludes that parthenium weed has the potential to be used as green manure for wheat.
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Many nucleic acid-based probe and PCR assays have been developed for the detection tracking of specific microbes within the rumen ecosystem [4-6, 8-11, 14, 15]. Conventional PCR assays detect PCR products at the end stage of each PCR reaction, where exponential amplification is no longer being achieved. This approach can result in different end product (amplicon) quantities being generated [3]. In contrast, using quantitative, or real-time PCR, quantification of the amplicon is performed not at the end of the reaction, but rather during exponential amplification, where theoretically each cycle will result in a doubling of product being created w [13]. For real-time PCR, the cycle at which fluorescence is deemed to be detectable above the background during the exponential phase is termed the cycle threshold (Ct). The Ct values obtained are then used for quantitation, which will be discussed later.
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Increased incidences of abiotic and biotic stresses impacting productivity in principal crops are being witnessed all over the world. Extreme events like prolonged droughts, intense rains and flooding, heat waves and frost damages are likely to further increase in future due to climate change. A wide range of adaptations and mitigation strategies are required to cope with such impacts. Efficient resource management and crop/livestock improvement for evolving better breeds can help to overcome abiotic stresses to some extent. However, such strategies being long drawn and cost intensive, there is a need to develop simple and low cost biological methods for the management of abiotic stress, which can be used on short term basis. Microorganisms could play a significant role in this respect, if we can exploit their unique properties of tolerance to extremities, their ubiquity, genetic diversity, their interaction with crop plants and develop methods for their successful deployment in agriculture production. Besides influencing the physico-chemical properties of rhizospheric soil through production of exopolysaccharides and formation of biofilm, microorganisms can also influence higher plants response to abiotic stresses like drought, chilling injury, salinity, metal toxicity and high temperature, through different mechanisms like induction of osmo-protectants and heat shock proteins etc. in plant cells. Use of these microorganisms per se can alleviate stresses in crop plants thus opening a new and emerging application in agriculture. These microbes also provide excellent models for understanding the stress tolerance, adaptation and response mechanisms that can be subsequently engineered into crop plants to cope with climate change induced stresses. KeywordsAbiotic stress tolerance–Microorganisms–Crop production–Climate change
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Identification of microorganisms by conventional methods requires the isolation of pure cultures followed by laborious characterization experiments. These procedures are therefore inadequate for study of the biodiversity of a natural or engineered ecosystem. A new set of molecular techniques developed during the 1990s revolutionized microbial ecology research. Among these techniques, cloning and the creation of a gene library, denaturant gradient gel electrophoresis (DGGE) and fluorescent in situ hybridization with DNA probes (FISH) stand out. Cloning provides very precise taxonomical information, but is time consuming and requires specialized personnel and so its introduction in wastewater treatment has been slow. DGGE is a rapid and simple method that provides characteristic band patterns for different samples, allowing quick sample profiling, while retaining the possibility of a more thorough genetic analysis by sequencing of particular bands. FISH makes possible to identify microorganisms at any desired taxonomical level, depending on the specificity of the probe used. It is the only quantitative molecular biology technique, although quantification is either complex or tedious and subjective. Combination with a confocal laser-scanning microscope allows the visualization of three-dimensional microbial structures (granules, biofilms). The methods discussed have deepened our understanding of the microbiology of biological wastewater treatment. PCR-based methods (cloning and DGGE) have proved suitable for identifying the microorganisms that form the sludge. Both DGGE and FISH have been extensively employed. FISH is currently being used for elucidation of the composition, quantification and distribution of different bacterial groups in granules and biofilms, as well as their structure and architecture.
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The bacterial diversity assessed from clone libraries prepared from rRNA (two libraries) and ribosomal DNA (rDNA) (one library) from polychlorinated biphenyl (PCB)-polluted soil has been analyzed. A good correspondence of the community composition found in the two types of library was observed. Nearly 29% of the cloned sequences in the rDNA library were identical to sequences in the rRNA libraries. More than 60% of the total cloned sequence types analyzed were grouped in phylogenetic groups (a clone group with sequence similarity higher than 97% [98% for Burkholderia and Pseudomonas-type clones]) represented in both types of libraries. Some of those phylogenetic groups, mostly represented by a single (or pair) of cloned sequence type(s), were observed in only one of the types of library. An important difference between the libraries was the lack of clones representative of the Actinobacteria in the rDNA library. The PCB-polluted soil exhibited a high bacterial diversity which included representatives of two novel lineages. The apparent abundance of bacteria affiliated to the beta-subclass of the Proteobacteria, and to the genus Burkholderia in particular, was confirmed by fluorescence in situ hybridization analysis. The possible influence on apparent diversity of low template concentrations was assessed by dilution of the RNA template prior to amplification by reverse transcription-PCR. Although differences in the composition of the two rRNA libraries obtained from high and low RNA concentrations were observed, the main components of the bacterial community were represented in both libraries, and therefore their detection was not compromised by the lower concentrations of template used in this study.
Since 2006 more than 50 Danish full-scale wastewater treatment plants with nutrient removal have been investigated in a project called 'The Microbial Database for Danish Activated Sludge Wastewater Treatment Plants with Nutrient Removal (MiDas-DK)'. Comprehensive sets of samples have been collected, analyzed and associated with extensive operational data from the plants. The community composition was analyzed by quantitative fluorescence in situ hybridization (FISH) supported by 16S rRNA amplicon sequencing and deep metagenomics. MiDas-DK has been a powerful tool to study the complex activated sludge ecosystems, and, besides many scientific articles on fundamental issues on mixed communities encompassing nitrifiers, denitrifiers, bacteria involved in P-removal, hydrolysis, fermentation, and foaming, the project has provided results that can be used to optimize the operation of full-scale plants and carry out trouble-shooting. A core microbial community has been defined comprising the majority of microorganisms present in the plants. Time series have been established, providing an overview of temporal variations in the different plants. Interestingly, although most microorganisms were present in all plants, there seemed to be plant-specific factors that controlled the population composition thereby keeping it unique in each plant over time. Statistical analyses of FISH and operational data revealed some correlations, but less than expected. MiDas-DK ( will continue over the next years and we hope the approach can inspire others to make similar projects in other parts of the world to get a more comprehensive understanding of microbial communities in wastewater engineering.
Various environmental conditions affecting total phosphorus removal from farm wastewater in a biofilm filter, process were investigated using loess balls andChromobacterium LEE-38 at a pilot plant. WhenChromobacterium LEE-38 was used, the removal efficiency of total phosphorous was approximately 10- or 5-fold higher than that ofAcinetobacter CHA-2-14 orAcinetobacter CHA-4-5, respectively. When a loess ball of 11–14 mm manufactured at a 960°C calcining temperature was used, the removal efficiency of total phosphorous was 90.0%. When 70% of the volume fraction was used, the maximum efficiency of total phosphorus removal was 93.1%. Notably, when the initial pH was in the range of 6.0 to 8.0, the maximum removal efficiency of total phosphorus was obtained after 30 days. When the operating temperature was in the range of 30 to 55°C, the maximum removal efficiencies of total phosphorus, 95.6 to 94.6%, were obtained. On the other hand, at operating temperatures below 20°C or above 40°C, the removal efficiency of total phosphorous decreased. Among the various processes, biofilm filter process A gave the highest removal efficiency of 96.4%. Pilot tests of total phosphorus removal using farm wastewater from the biofilm filter process A were carried out for 60 days under optimal condition. WhenAcinetobacters sp. Lee-11 was used, the average removal efficiency in thep-adsorption area was only 32.5%, and the removal efficiencies of chemical oxygen demand (COD) and biological oxygen demand (BOD) were 56.7 and 62.5%, respectively. On the other hand, whenChromobacterium LEE-38 was used, the average removal efficiency was 95.1%, and the removal efficiencies of COD and BOD were 91.3 and 93.2%, respectively.
The combination of PCR amplification of 16S rRNA genes with denaturing gradient gel electrophoresis (DGGE) analysis was used to reveal the compositions and dynamics of bacterial communities in a sewage treatment plant with two systems, i.e., an anoxic-anaerobic-aerobic system (inverted A20) and an anaerobic-anoxic-aerobic one (conventional A20) over a period from February to July 2009, during which both systems experienced serious sludge bulking problems. The DGGE patterns showed that there were many common bands in both systems, suggesting the high similarity of bacterial communities of the two systems. Meanwhile, the moving window correlation analysis showed that the two systems experienced different microbial community structure changes during the period, which might be related with the different situations of the occurrence and disappearance of sludge bulking, as being reflected by sludge volume index (SVI) values. Major bands of DGGE patterns of sludge samples were further sequenced. Phylogenetic affiliation indicated that the majority of the sequences obtained were affiliated with Actinobacteria, Firmicutes, Bacteroidetes/Chlorobi group and alpha- and beta-Proteobacteria. Two sequences showed high similarities to typical filamentous bacteria Microthrix parvicella and Nostocoida limicola I, indicating that these bacterial species have been involved in the sludge bulking problems.