\r\n\t1. Geopolymers chemistry topic describes the chemical reaction models and chemical kinetic of the geopolymerization which occurs after mixing the aluminosilicate raw materials with an alkaline solution.
\r\n\t2. Advanced characterization of geopolymers topic includes innovative technologies applied on geopolymers characterization at the nanoscale level, meant to explain the bond between the reacted and nonreacted particles from the composition.
\r\n\t3. Sustainability with geopolymers topic should provide clear information about the characteristics and applications of the geopolymers which use as raw materials industrial waste. Moreover, environmental impact studies which offer a clear view of the effects produced by geopolymers manufacturing, compared to conventional materials, is included.
\r\n\t4. Geopolymers as functional materials topic will present key aspects in developing geopolymers with tailored properties that increase further the heavy metals adsorption capacity, offering outstanding opportunities for energy-efficient separations and process intensification, in terms of saving energy, reducing capital costs, minimizing environmental impact and maximizing the raw materials exploitation.
\r\n\t5. Reinforced structures topic describe the effects produced by the introduction, in the geopolymers matrix, of different types of reinforcing elements.
It is well known that the diet fed to dairy cows is an important lever by which milk yield and milk composition could be modified. Although milk production is affected by numerous dietary nutrients, energy and protein are most critical. Feed grains containing starch such as corn, wheat, barley, and sorghum, as a primary source of energy, are commonly fed to livestock to improve meat or milk productions [1]. Improving starch utilization may improve lactation performance in cows and reduce feed costs, especially when grain price is high [2]. Whole grain with an intact pericarp is largely or entirely resistant to digestion by ruminants because whole kernels are resistant to bacterial and host enzyme accessing to endosperm of grain kernel in the rumen and in the intestine, respectively [3, 4]. Therefore, cereal grains require processing to break the protective seed coat, and the fibrous hull in the case of barley and oats, and to improve grain digestibility [5]. Grain can be processed by the application of various combinations of heat, moisture, time, and mechanical action. Nonthermal processes (roller and hammer mill) and thermal processes (roasting, popping, micronizing, autoclaving, steam-flaking, steam pelleting, expanding, extruding, and toasting) could be used to manipulate rate of degradation and hence ruminal availability [6, 7]. The extent and rate of ruminal digestion of grain can be manipulated through processing [8]. The quality of processed grain can be affected by the application of various combinations of heat, moisture, time, and mechanical action [9]. These processing treatments alter kernel structure, thereby enhancing the release of starch granules from the protein matrix and disrupting their order during gelatinization; this, in turn, increases the accessibility of starch to microbes in the rumen and increases the susceptibility to enzyme activity [10]. Thus, grain processing can be a useful tool for optimizing lactating dairy cow production by synchronizing energy and protein to improve rumen microbial protein production. Steam-flaking is a more extensive processing system than dry- or steam-rolling [11]. Steam-flaking has been shown to increase the digestibility of starch by cattle fed corn- or sorghum grain-based diets over whole, ground, or dry-rolled grain [7]. In sorghum, steam-flaking has been found to increase ruminal starch digestion, as compared with dry-rolling or grinding [12]. Consistent increases in ruminal starch digestion have also been observed for steam-flaked corn, as compared with ground or dry-rolled corn [13]. Malcolm and Kiesling [14] reported that steam-flaking of barley tended to increase in situ rumen dry matter degradation. These experiments have shown that steam-flaking increases the amount of starch fermented in the rumen and, also, enhances the intestinal digestibility of starch that escape the rumen degradation. Few studies have investigated the effects of steam-flaked barley (SFB) on its digestibility in the rumen and in the total digestive tract of lactating dairy cows. Increasing the extent of grain processing enhances ruminal starch digestion of grain [8]. Additionally, grinding is a conventional processing method used in grain processing for feeding dairy cattle in some countries like Iran because of its low cost, but ground barley is often dusty, thereby potentially reducing feed intake. This response is usually influenced by either the extent of processing or flake density [13]. The objective of this chapter was to discuss the effects of grain processing methods with emphasizes on grinding and steam-flaking and on grain digestibility and lactation performance of dairy cows.
Cereal grains are rich in starch ranging from 40% in oats up to 80% of dry matter in rice, with the variation in starch content dependent on variety, climatic conditions, and agronomic practices [15]. Starch is synthesized into a form of rough spherical granules and, within each feed grain starch granule, multiple concentric semicrystalline and amorphous shells are present [1]. Feed grain starch consists of amylose and amylopectin, as two major components [1, 5], which are present in different proportions in the starch granule of feed grains [1]. Cereal starches are typically composed of 25–28% amylose and 72–75% amylopectin. There are also waxy cultivars with very high amylopectin concentration (up to 100%) and high amylase (up to 70% amylose) cultivars [5]. Starch of waxy barley varieties may be less digestible than normal barley due to the differences in chemical composition and starch structure. Starch granules isolated from different feed grains and other starch sources reveal characteristic morphologies, varying in shape [16], in molecular size [17], and in specific surface structure and porosity [18]. Also, it has been recognized that cultivars or varieties in feed grains vary in granule size distribution, suggesting a significant genetic control [19]. Other components of feed grains, including lipids, proteins, and minerals, are also associated with starch granules. Lipids and proteins are the most abundant nonstarch components in feed grains that may affect physical state and enzyme susceptibility of starch in livestock [20].
Whole grains with an intact pericarp are largely or entirely resistant to digestion by ruminants because whole kernels are resistant to bacterial attachment in the rumen [3, 4]. Unlike sheep, adult cattles have a limited capacity to masticate cereal grains; hence, it is essential to break the pericarp of the seed either through chemical or physical treatments [21]. In feed grains, germ and endosperm are surrounded by the pericarp, which is largely resistant to microbial attachment [22]. Starch granules from corn grain are surrounded by a protein matrix in the endosperm [23], which influences digestion by microorganisms [24]. Feed grain endosperm encapsulated by protein matrix acts as a physical barrier to protect from enzymatic hydrolysis [25]. It has been shown that this protein matrix by blocking the absorption sites or by influencing enzyme binding may reduce the surface availability of starch to host enzyme and ruminal bacteria [25]. In addition, the results of several studies have shown that hydrophobic properties of grain protein matrix, associated with type and location of proteins, could be responsible for the differences in starch digestion between rapidly digested grains such as wheat, barley, and rice, and slowly digested grains such as maize and sorghum [2, 24]. Processing is necessary for feed grains to break the protective seed coat [3, 5] especially for the grains such as maize and sorghum [11], and the fibrous hull in case of barley and oats, to improve their digestibility in the digestive tract of animals [5]. Feed grain processing by grinding, rolling, pelleting, steam-rolling, or steam-flaking breaks down the recalcitrant barriers such as the hull, pericarp, and protein matrix and allows microbial and host enzyme accessing to the starch within endosperm cells.
Grain processing methods could be divided into two groups: (1) nonthermal processes such as roller and hammer mill and (2) thermal processes, which include dry processing (roasting, popping, and micronizing) and wet processing (autoclaving, steam-flaking, steam-pelleting, expanding, extruding, and toasting) [26]. Heat processing, however, has been associated with increased efficiency of fermentative utilization by altering the protein matrix of the endosperm and the starch structure, thus allowing a better utilization by microbial enzymatic digestion [21]. Increasing the starch availability is the primary goal of grain processing. In addition, processing may destroy mycotoxins and improve mixing characteristics to improve bunk management and thereby enhance animal performance [27]. The processes reduce the particle size of the grain, increasing the surface area available for microbial attachment and colonization; combined, these actions increase the rate and extent of starch digestion [22]. Starch gelatinization, a process in which disaggregated amylose and amylopectin chains in a gelatinized starch paste reassociate to form more ordered structures, and dextrination, formation of dextrins (fragments of amylose and amylopectin molecules formed by heating dry starch in the presence of some moisture, acids, or salts) during processing of grains [28], improve accessibility of enzymes to the starch granules. It, in turn, may shift the site of digestion of protein and starch from the rumen to the intestine [6, 7], and consequently, it results in an improved supply of amino acids and glucose to animal metabolism [29]. Elevated glucose absorption represents one mechanism by which increased intestinal starch digestion might increase milk yield [30]. Increased net energy density of cereals is also beneficial because high-yield dairy cows often are unable to consume sufficient energy during early lactation to meet the requirements.
Ruminants do not produce appreciable quantities of salivary amylase, and therefore, starch digestion in ruminants is initiated after ingestion and mastication of feed [1]. Dairy cattle consume large amounts of starch (20–40% of diet DM) as a way to increase energy consumption in support of high milk production [31]. The optimum starch content of diets for lactating dairy cows is not well defined, but 24–26% starch (dry matter basis) has been suggested [32]. Total tract starch digestibility of dairy cows is highly variable and has a range of 70–100% [33]. Quantitatively, most of the starch digestion in ruminants [2] occurs in the rumen (55–85%) and the small intestine (15–25%) with indigestible starch fractions ranging from 0 to 20% of starch intake [2, 34]. The rate and extent of starch digestion in the rumen are determined by interrelations among several factors, including level of feed intake, source of dietary starch, diet composition, grain processing, chemical alterations (fermentation, gelatinization), grain passage rate, and degree of adaptation of rumen bacteria to dietary starch [35]. Digestion of starch in the rumen is facilitated symbiotically by ruminal bacteria with amylolytic capacity [35]. Attachment of bacteria to starch containing feed particles is a key step in bacterial fermentation of starch in the rumen [1]. Although ruminal protozoa play a role in ruminal starch digestion through ingesting and digesting starch granules, eliminating protozoa by rumen defaunation can actually increase rate and extent of ruminal starch digestion [36], which suggests that bacterial fermentation alone is sufficient in starch digestion in the rumen. In general, ruminal starch digestion can vary substantially, ranging from 40 to 80% in lactating dairy cows [2, 33]. Compared to beef cattle, the extent of ruminal starch digestion from maize grain is often lower for lactating dairy cows [34]. The high levels of feed intake, rapid rates of passage and minimal mastication of grain particles by lactating dairy cows are thought to be responsible for the lower ruminal starch digestion [33, 34]. Approximately 50–90% of the postruminal flow of starch is digested in the small intestine [1, 3] by enzymatic hydrolysis of starch that provides energy in the form of glucose to the animal. The passage rate through the small intestine, surface exposure, and grain endosperm properties can alter the output of enzymatic starch hydrolysis in the small intestine [37]. In the large intestine, around 30–60% of the starch escaping ruminal digestion and enzymatic hydrolysis can be digested via hind gut bacterial fermentation [6] that is symbiotically providing energy in the form of volatile fatty acid.
The kinetics of starch digestion in livestock animals depend largely on two major factors: (1) the inherent starch architecture and related physicochemical properties and (2) the degree of processing feed grains prior to feeding that vary with methods used such as rolling, pelleting, flaking, extrusion, and expander processing as well as the processing condition setting up of each method [1]. Starches from various sources show different responses to the heat processing conditions. The starch structure can be modified by the application of such processing, thus some native physicochemical starch properties including starch granule morphology, crystallinity, the amylose content, and the type of endosperm are potentially affected [38]. These native physicochemical properties have been recognized to influence the starch digestion of grains [38]. Feed grains fed to livestock are commonly processed through a grinder or a roller prior to feeding, and mechanical processing could be considered as one of the primary and less expensive methods to influence starch digestibility. In commercial sector, various processing equipments with variable screen sizes or mill settings are available to create wide distributions of grain particle sizes and to meet different requirements of animal feeding [39]. The effect of particle size distribution on starch digestion is primarily associated with the available surface area for microbial access and enzymatic hydrolysis [40]. The particle size of grains after processing, even if not directly related to native physiochemical starch properties, also plays a role in determining the relative efficiency of starch digestion within grains [41]. Blasel et al. [41] reported a reduction in the degree of starch access by α-amylase of about 27 g/kg of starch for each 100 mm increase in particle size in ground maize grain. Also, it is reported a reduction in the rate of starch digestion of grounded feed grains through 3.0 mm compared with 0.8 mm opening screen after either 60 or 240 min of in vitro incubation [42]. In addition, it has been observed that there is an inverse square relationship between the enzymatic hydrolysis rates of pancreatic α-amylase and the increasing particle size in milled barley and sorghum grains [43].
\nThe method of processing grain will depend on the type of grain and the specific feeding and management conditions. Small kernel grain, such as barley and wheat, is usually processed by dry-rolling, steam-rolling, or temper-rolling, while corn grain is often steam-flaked in cattle feeding. Grain can also be treated with chemicals or enzymes to alter the rate and extent of nutrient degradation [26]. Mechanical processing by rolling of the grain cracks or crushes the fibrous hull and pericarp to enable access of rumen microorganisms and enzymes to the internal endosperm and increases the surface area for attachment [22]. The addition of moisture in steam-rolling and temper-rolling may be advantageous over dry-rolling when the original grain is very dry (<10% moisture), of variable kernel size, rolled more extensively to maximize utilization, and fed with limited amount of forage. Steam-processed grain is exposed to steam at either atmospheric, low, or high pressure to increase grain moisture and temperature before the grain is rolled. Steam-flaking of grain is a more extensive process with longer steam conditioning times and thinner rolled flakes than steam-rolling. The combination of moisture, heat, and rolling causes gelatinization of the starch granules, i.e., swelling of granules as water is absorbed, disruption of the crystalline structure, dissolution of polysaccharides, and diffusion from ruptured granules. Steam-flaking of barley grain increased the in vitro amyloglucosidase, catalyzed release of glucose and the rate of starch degradation compared to dry-rolling of the grain [44]. However, the beneficial effects of starch gelatinization for barley may be less than for corn or sorghum grain because barley starch once it becomes accessible readily degraded by ruminal microorganisms [44].
The optimal degree of grain processing is aimed at achieving a balance between maximizing the extent and controlling the rate of starch digestion in the rumen to increase utilization and avoid digestive and metabolic disturbances [5]. The degree of grain processing, also called the extent of processing, can significantly affect rate and extent of grain digestibility in the rumen or in the total digestive tract, thus affect feeding value and cattle performance as well as cattle health when the high-grain is fed. Overprocessed grain increases proportion of fine particles, reduces palatability, increases rate of grain digestion in the rumen, hence increases the incidence of digestive disorders such as rumen acidosis, bloat, laminitis, and liver abscesses [5]; whereas underprocessed grain reduces the starch availability for fermentation by rumen microbes [45]. The degree of grain processing has been widely used by cattle nutritionist to manipulate rate and extent of grain digestion in the rumen. Processing index was developed by our laboratory to quantify the degree of processing, and it is currently applied as a routine method by feed mill in grain processing for cattle feeding [4]. The processing index refers to the volume weight (g/L) of barley after processing expressed as a percentage of its volume weight before processing [4]. This index reflects the fact that the more extensively grain is processed (i.e., the higher the degree of processing), the finer the particle will be, hence, the lower the volume weight will be, and consequently, the lower the processing index. Take barley grain as an example; the disadvantages of under- or overprocessing grain on the milk production of dairy cattle were described in a study by Yang et al. [4]; barley grain was steam-rolled to four degrees with processing index of 81, 72, 64, and 55%, and milk production increased quadratically with degree of processing resulting, respectively, in increases of 0, 9.8, 20.3, and 13.3% in milk yield. The reduced milk yield in dairy cows fed coarsely rolled barley grain compared to those fed more extensively processed barley was due to the combined effects of lower feed intake, lower nutrient digestibility, and a slower particle outflow rate from the rumen. We concluded that the optimum extent of barley processing for dairy cows fed diets supplying adequate fiber was a processing index of 64% to maximize milk yield without negatively affecting milk fat percentage.
Kernel uniformity of grains can vary considerably depending on variety, growing conditions, disease, etc., in particular, grains with lower and higher volume weights are often blended commercially to achieve an intermediate volume weight that increases the market value of the grain [5], but it also increases the variability of kernel size. The poor kernel uniformity can contribute to inconsistent processing especially when the dry-rolling technique is used. We have developed the precision processing method [45–47] that refers to separate fractions of uniform kernel size and then each fraction is processed with an optimum roller setting specific to the kernel size to ensure that all kernels are cracked [46]. For barley grain of variable kernel size and shape, precision processing enables greater control over the extent of processing compared with processing with the conventional one roller setting, particularly when the grain is dry-rolled. Precision processing increased digestibility of crude protein and acid detergent fiber; as a result, organic matter digestibility increased as compared with the conventional processing in a study using beef cattle fed high-grain diet [46]. The barley that was sieved and rolled with settings based on kernel size had increased starch degradability up to 24 h and increased overall rate and effective degradability of starch in the rumen compared to the barley processed as a single roller setting [45]. In a lactation study, dry matter intake, ruminal fermentation, nutrient digestibility, and milk production were not affected by precision processing compared with conventional processing [47]. The differences in the results between the beef and dairy studies are likely due to the differences in the amount of barley grain (67 versus 40%) and forage (10 versus 40%) in the diets.
Compared with grinding, rolling processing generally reduces starch digestibility in the small intestine and in the total digestive tract of cattle [26]. Larsen et al. [48] in a review reported that reduction in ruminal starch digestion of grains was generally not compensated for by increasing the small intestinal starch digestion but was associated with increases of starch digestion in hindgut. Santos et al. [49] reported that dairy cows fed steam-flaked corn yielded daily 1.5 kg more milk than did cows fed rolled grain, also the yield of milk protein and percentages of lactose and solids that are nonfat increased compared with rolling processing. Similarly, compared with rolling, feeding steam-flaked corn and sorghum increased yields of milk, milk protein, and fat, and protein percentage of milk, which were likely due to the increased total tract digestibility of dry matter, crude protein, starch, and neutral detergent fiber [50]. It suggests that steam-flaking versus rolling improved the feeding value of corn for lactating cows by improving diet acceptability and digestibility of organic matter and increased the estimated net energy for lactation of corn. López-Soto et al. [51] indicated that compared to dry-rolled barley, steam-flaked barley increased ruminal digestion of organic matter and starch of diet, but decreased dry matter intake of lactating dairy cows. Our in situ experiment [52] showed that ground barley versus steam-flaked barley increased organic matter disappearance (Figure 1) and increased the washable fraction (28.3 versus 21.4%), rate of potentially degradable fraction (0.10 versus 0.05 h−1), and effective degradability (60.6 versus 47.6%) of organic matter. In an in vivo study [53] using lactating dairy cows, we observed that the grinding versus steam-flaking barley grain did not affect dry matter intake (23.6 kg/day), digestibility of dry matter in the total digestive tract (71.0%), milk yield (43.4 kg/day), milk components, rumen pH, and molar proportions of acetate, propionate, and butyrate. The lack of difference between grinding and flaking could be due to low grain inclusion rate and minimal difference in the particle size as barley was coarsely ground. In contrast, Eastridge et al. [54] reported that the finely ground corn decreased milk urea nitrogen content compared with coarsely ground corn, suggesting that finely ground corn provided more fermentable starch in the rumen, thus possibly improved bacterial capture of the nitrogen. However, the results on dry matter intake, milk production of lactating dairy cows by feeding steam-flaked corn versus ground corn are inconsistent [55–57]. In another study using lactating dairy cows, we found that grinding versus steam-flaking of barley affected dry matter intake, organic matter digestibility in the total digestive tract, and feed efficiency. The discrepancy among the studies could be due to differences in grain variety, particle size distribution of processed grain, inclusion rate of grain in the diets, which could affect the grain digestibility in the rumen and in the intestine, hence, impact feed intake and milk production of dairy cows (Figure 2).
Organic matter disappearance (%) of ground barley (GB) compared to steam-flaked barley (SFB).
Organic matter disappearance (%) of steam-flaked barley (SFB) with various processing indexes (%).
Flake density of grains affects grain digestibility and cow performance. It was suggested that optimal flake density was between 0.32 and 0.39 kg/L [58]. Zinn and Barajas [59] conducted an experiment to evaluate the influence of flake density on feeding value of a barley-corn blend fed to feedlot cattle, and observed an increase of ruminal digestibility of organic matter, starch, and protein, but a decrease of ruminal nitrogen efficiency with decreasing flake density. We found that decreasing the density of steam flaked barley from 0.30 to 0.26 kg/L increased ruminal digestion of starch and ruminal propionate, but decreased dry matter intake and ruminal protein degradation. These results are expected since decreasing the density of processed barley indicated increased degree of processing, hence increased rumen starch digestibility (Figure 2), and consequently, decreased rumen pH (potential rumen acidosis), and decreased dry matter intake. In comparing with corn-based diet, dairy cows fed barley-based diets showed greater dietary energy due to improved ruminal microbial efficiency, greater total tract organic matter digestion, and lower ruminal acetate and methane production. However, dairy cows that were fed barley-based diets had lower ruminal pH, which was exacerbated as flake density decreased. Results showed 8% improvement in energy value of barley with steam-flaking [51]. Flaking barley too thinly would depress feed intake because of increase in rumen digestion and reduction in rumen pH [51]. Our study also showed that decreasing the density of steam-flaked barley from 390 to 340 to 290 g/L tended to linearly decrease dry matter intake, total solids percentage of milk, and linearly decreased milk urea nitrogen. Finely ground corn increased ruminal propionate concentration and decreased ruminal pH and acetate to propionate ratio, suggesting an increase of grain digestion in the rumen. Increasing density of steam-flaked corn increased total tract digestion of organic matter, neutral detergent fiber, starch, and digestible energy content of diet and increased milk efficiency [60]. It suggests that barley and corn should be processed at different density when it is steam flaked to maximize its digestibility, while to minimize its rumen health problem since barley is more rapidly digestible than corn in the rumen.
Improvement and optimization of grain digestion in the rumen and the intestine is an important research focus in cattle nutrition and feeding. Generally, site, extent, and rate of grain starch digestion in the digestive tracts of cattle are influenced by intrinsic and external factors that can be interrelated, hence they are not easily defined. Particle size reduction, starch gelatinization, retrogradation, and dextrination due to grain processing may shift the site of starch digestion from the rumen to the intestine, and thus it results in an improved supply of amino acids and glucose to animal. Steam-flaking grain increases the starch digested both in the rumen and in the intestine; this, in turn, increases the available energy for milk production. Overall, both ground and steam-flaked grains could be fed dairy cows depending on the level of grain in diet, dietary composition, and economic cost of grain processing.
\nFeeding values differ among grain sources and processing methods used. Although the net energy value of grain usually is increased by more extensive flaking, regardless of grain source or processing, dairy cows can produce milk at similar rates, probably due to reduction in dry matter intake and lower passage rate of flaked grains compared with ground grains. This may support the concept that chemostatic factors generally control intake of low forage diets. For a specific grain and processing method, roughage source and moisture may markedly influence rate of production and net energy value of the grain, probably due to bunk management, diet acceptability, chewing activity, and site and extent of starch digestion.
Bioremediation and natural reduction are also seen as a solution for emerging contaminant problems; microbes are very helpful to remediate the contaminated environment. Number of microbes including aerobic, anaerobic bacteria and fungi are involved in bioremediation process. Bioremediation is highly involved in degradation, eradication, immobilization, or detoxification diverse chemical wastes and physical hazardous materials from the surrounding through the all-inclusive and action of microorganisms. The main principle is degrading and converting pollutants to less toxic forms. There are two types of factors these are biotic and abiotic conditions are determine rate of degradation. Currently, different methods and strategies are applied for bioremediation process.
Environmental pollution has been on the rise in the past few decades due to increased human activities such as population explosion, unsafe agricultural practices, unplanned urbanization, deforestation, rapid industrialization and non-judicious use of energy reservoirs and other anthropogenic activities. Among the pollutants that are of environmental and public health concerns due to their toxicities are: chemical fertilizer, heavy metals, nuclear wastes, pesticides, herbicides, insecticides greenhouse gases, and hydrocarbons. Thousands of hazardous waste sites have been identified and estimated is that more will be identified in the coming decades. Release of pollutants into the environment comes from illegal dumping by chemical companies and industries. Many of the techniques utilized for site clean-up in the past, such as digging up the contaminated soil and hauling it away to be land filled or incinerated have been prohibitively expensive and do not provide permanent solution. More recent techniques such as vapor extraction and soil venting are cost effective but incomplete solution.
Bioremediation is a process where biological organisms are used to remove or neutralize an environmental pollutant by metabolic process. The “biological” organisms include microscopic organisms, such as fungi, algae and bacteria, and the “remediation”—treating the situation.
In the Earth’s biosphere, microorganisms grow in the widest range of habitats. They grow in soil, water, plants, animals, deep sea, and freezing ice environment. Their absolute numbers and their appetite for a wide range of chemicals make microorganisms the perfect candidate for acting as our environmental caretakers.
“Bioremediation is a waste management technique that includes the use of living organisms to eradicate or neutralize pollutants from a contaminated site.”
“Bioremediation is a ‘treatment techniques’ that uses naturally occurring organisms to break down harmful materials into less toxic or non-toxic materials.”
Bioremediation technologies came into extensive usage and continue growing today at an exponential rate. Remediation of polluted sites using microbial process (bioremediation) has proven effective and reliable due to its eco-friendly features. In the past two decades, there have been recent developments in bioremediation techniques with the decisive goal being to successfully restore polluted environments in an economic, eco-friendly approach. Researchers have developed different bioremediation techniques that restore polluted environments. The micro-organisms used in bioremediation can be either indigenous or non-indigenous added to the contaminated site. Indigenous microorganisms present in polluted environments hold the key to solving most of the challenges associated with biodegradation and bioremediation of pollutant [1]. Environmentally friendly and cost effective are among the major advantages of bioremediation compared to both chemical and physical methods of remediation.
A mechanism of bioremediation is to reduce, detoxify, degrade, mineralize or transform more toxic pollutants to a less toxic. The pollutant removal process depends mainly on the pollutant nature, which includes pesticides, agrochemicals, chlorinated compounds, heavy metals, xenobiotic compounds, organic halogens, greenhouse gases, hydrocarbons, nuclear waste, dyes plastics and sludge. Cleaning technique apply to remove toxic waste from polluted environment. Bioremediation is highly involved in degradation, eradication, immobilization, or detoxification diverse chemical wastes and physical hazardous materials from the surrounding through the all-inclusive and action of microorganisms (Figure 1).
Bioremediation approaches for environmental clean-up.
Microorganisms play an important role on nutritional chains that are important part of the biological balance in life. Bioremediation involves the removal of the contaminated materials with the help of bacteria, fungi, algae and yeast. Microbes can grow at below zero temperature as well as extreme heat in the presence of hazardous compounds or any waste stream. The two characters of microbes are adaptability and biological system made them suitable for remediation process [2]. Carbon is the main requirement for microbial activity. Bioremediation process was carried out by microbial consortium in different environments. These microorganisms comprise Achromobacter, Arthrobacter, Alcaligenes, Bacillus, Corynebacterium, Pseudomonas, Flavobacterium, Mycobacterium, Nitrosomonas, Xanthobacter, etc. [3].
There are groups of microbes which are used in bioremediation such as:
Aerobic: aerobic bacteria have degradative capacities to degrade the complex compounds such as Pseudomonas, Acinetobacter, Sphingomonas, Nocardia, Flavobacterium, Rhodococcus, and Mycobacterium. These microbes have been reported to degrade pesticides, hydrocarbons, alkanes, and polyaromatic compounds. Many of these bacteria use the contaminants as carbon and energy source.
Anaerobic: anaerobic bacteria are not as regularly used as aerobic bacteria. There is an increasing interest in aerobic bacteria used for bioremediation of chlorinated aromatic compounds, polychlorinated biphenyls, and dechlorination of the solvent trichloroethylene and chloroform, degrading and converting pollutants to less toxic forms.
Bioremediation process is degrading, removing, changing, immobilizing, or detoxifying various chemicals and physical pollutants from the environment through the activity of bacteria, fungi, algae and plants. Enzymatic metabolic pathways of microorganisms facilitate the progress of biochemical reactions that help in degradation of the pollutant. Microorganisms are act on the pollutants only when they have contact to the compounds which help them to generate energy and nutrients to multiply cells. The effectiveness of bioremediation depends on many factors; including, the chemical nature and concentration of pollutants, the physicochemical characteristics of the environment, and their accessibility to existing microorganisms [4].
The factors are mainly microbial population for degrading the pollutants, the accessibility of contaminants to the microbial population and environment factors like type of soils, pH, temperature, oxygen and nutrients.
Biotic factors are helpful for the degradation of organic compounds by microorganisms with insufficient carbon sources, antagonistic interactions among microorganisms or the protozoa and bacteriophages. The rate of contaminant degradation is frequently dependent on the concentration of the contaminant and the amount of catalyst present in biochemical reaction. The major biological factors are included enzyme activity, interaction (competition, succession, and predation), mutation, horizontal gene transfer, its growth for biomass production, population size and its composition [5, 6].
The interaction of environmental contaminants with metabolic activity, physicochemical properties of the microorganisms targeted during the process. The successful interaction between the microbes and pollutant depends on the environmental situations. Microbial growth and activity are depended on temperature, pH, moisture, soil structure, water solubility, nutrients, site conditions, oxygen content and redox potential, deficiency of resources and physico-chemical bioavailability of pollutants, concentration, chemical structure, type, solubility and toxicity. This above factors are control degradation kinetics [5, 7].
Biodegradation of pollutant can occur under range of pH (6.5–8.5) is generally optimal for biodegradation in most aquatic and terrestrial environment. Moisture affects the metabolism of contaminant because it depends on the kind and amount of soluble constituents that are accessible as well as the pH and osmotic pressure of terrestrial and aquatic systems [8].
Superficially, bioremediation techniques can be carried out ex-situ and in-situ site of application (Figure 1). Pollutant nature, depth and amount of pollution, type of environment, location, cost, and environmental policies are the selection standards that are considered for selecting any bioremediation technique. Performance based on oxygen and nutrient concentrations, temperature, pH, and other abiotic factors that determine the success of bioremediation processes [9, 10].
Ex-situ bioremediation techniques involve digging pollutants from polluted sites and successively transporting them to another site for treatment. Ex-situ bioremediation techniques are regularly considered based on the depth of pollution, type of pollutant, degree of pollution, cost of treatment and geographical location of the polluted site. Performance standards also regulate the choice of ex-situ bioremediation techniques.
Solid-phase treatment
Solid-phase bioremediation is an ex-situ technology in which the contaminated soil is excavated and placed into piles. It is also includes organic waste like leaves, animal manures and agriculture wastes, domestic, industrial wastes and municipal wastes. Bacterial growth is moved through pipes that are distributed throughout the piles. Air pulling through the pipes is necessary for ventilation and microbial respiration. Solid-phase system required huge amount of space and cleanups require more time to complete as compared to slurry-phase processes. Solid-phase treatment processes include biopiles, windrows, land farming, composting, etc. [11].
Slurry-phase bioremediation
Slurry-phase bioremediation is a relative more rapid process compared to the other treatment processes. Contaminated soil is combined with water, nutrient and oxygen in the bioreactor to create the optimum environment for the microorganisms to degrade the contaminants which are present in soil. This processing involves the separation of stones and rubbles from the contaminated soil. The added water concentration depends on the concentration of pollutants, the biodegradation process rate and the physicochemical properties of the soil. After completion of this process the soil is removed and dried up by using vacuum filters, pressure filters and centrifuges. The subsequent procedure is soil disposition and advance treatment of the resultant fluids.
There are far more than nine types of bioremediation, but the following are the most common ways in which it is used.
Bioremediation includes above-ground piling of dug polluted soil, followed by aeration and nutrient amendment to improve bioremediation by microbial metabolic activities. This technique comprises aeration, irrigation, nutrients, leachate collection and treatment bed systems. This specific ex-situ technique is progressively being measured due to its useful features with cost effectiveness, which allows operative biodegradation conditions includes pH, nutrient, temperature and aeration are effectively controlled. The biopile use to treat volatile low molecular weight pollutants; it can also be used effectively to remediate polluted very cold extreme environments [12, 13, 14]. The flexibility of biopile allows remediation time to be shortened as heating system can be integrated into biopile design to increase microbial activities and contaminant availability thus increasing the rate of biodegradation [15]. Additionally, heated air can be injected into biopile design to deliver air and heat in tandem, in order to facilitate enhanced bioremediation. Bulking agents such as straw saw dust, bark or wood chips and other organic materials have been added to enhance remediation process in a biopile construct. Although biopile systems connected to additional field ex-situ bioremediation techniques, such as land farming, bioventing, biosparging, robust engineering, maintenance and operation cost, lack of power supply at remote sites, which would facilitate constant air circulation in contaminated piled soil through air pump. Additional, extreme heating of air can lead to soil drying undertaking bioremediation, which will inhibit microbial activities and which stimulate volatilization than biodegradation [16].
Windrows is bioremediation techniques depends on periodic rotating the piled polluted soil to improve bioremediation by increasing microbial degradation activities of native and transient hydrocarbonoclastic present in polluted soil. The periodic turning of polluted soil increase in aeration with addition of water, uniform distribution of nutrients, pollutants and microbial degradation activities, accordingly increase the rate of bioremediation, which can be proficient through acclimatization, biotransformation and mineralization. Windrow treatment as compared to biopile treatment, showed higher rate of hydrocarbon removal however, the effectiveness of the windrow for hydrocarbon removal from the soil [17]. However, periodic turning associated with windrow treatment not the best selection method to implement in bioremediation of soil polluted with toxic volatiles compounds. The use of windrow treatment has been associated in greenhouse gas (CH4) release due to formation of anaerobic zone inside piled polluted soil, which frequently reduced aeration [18].
Land farming is the simplest, outstanding bioremediation techniques due to its low cost and less equipment requirement for operation. It is mostly observed in ex-situ bioremediation, while in some cases of in-situ bioremediation technique. This consideration is due to the site of treatment. Pollutant depth is important in land farming which can be carried out ex-situ or in-situ. In land farming, polluted soils are regularly excavated and tilled and site of treatment speciously regulates the type of bioremediation. When excavated polluted soil is treated on-site, it is ex-situ as it has more in common than other ex-situ bioremediation techniques. Generally, excavated polluted soils are carefully applied on a fixed layer support above the ground surface to allow aerobic biodegradation of pollutant by autochthonous microorganisms [19]. Over all, land farming bioremediation technique is very simple to design and implement, requires low capital input and can be used to treat large volume of polluted soil with minimal environmental impact and energy requirement [20].
Bioreactor is a vessel in which raw materials are converted to specific product(s) following series of biological reactions. There are different operational modes of bioreactors, which include: batch, fed-batch, sequencing batch, continuous and multistage. Bioreactor provides optimal growth conditions for bioremediation. Bioreactor filled with polluted samples for remediation process. The bioreactor based treatment of polluted soil has several advantages as compared to ex-situ bioremediation procedures. Bioreactor-based bioremediation process having excellent control of pH, temperature, agitation and aeration, substrate and inoculum concentrations efficiently reduces bioremediation time. The ability to control and manipulate process parameters in a bioreactor implies that biological reactions. The flexible nature of bioreactor designs allows maximum biological degradation while minimizing abiotic losses [21].
Advantages of ex-situ bioremediation
Suitable for a wide range of contaminants
Suitability relatively simple to assess from site investigation data
Biodegradation are greater in a bioreactor system than or in solid-phase systems because the contaminated environment is more manageable and more controllable and predictable.
Disadvantages
Not applicable to heavy metal contamination or chlorinated hydrocarbons such as trichloroethylene.
Non-permeable soil requires additional processing.
The contaminant can be stripped from soil via soil washing or physical extraction before being placed in bioreactor.
These techniques comprise treating polluted substances at the pollution site. It does not need any excavation and by little or no disturbance in soil construction. Perfectly, these techniques should to be cost effective compared to ex-situ bioremediation techniques. Some in-situ bioremediation techniques like bioventing, biosparging and phytoremediation may be enhanced, while others may be progress without any form of improvement such as intrinsic bioremediation or natural attenuation. In-situ bioremediation techniques have been effectively used to treat chlorinated solvents, heavy metals, dyes, and hydrocarbons polluted sites [22, 23, 24].
In-situ bioremediation is two types; these are intrinsic and engineered bioremediation.
Intrinsic bioremediation
Intrinsic bioremediation also known as natural reduction is an in-situ bioremediation technique, which involves passive remediation of polluted sites, without any external force (human intervention). This process deals with stimulation of indigenous or naturally occurring microbial population. The process based on both microbial aerobic and anaerobic processes to biodegrade polluting constituents containing those that are recalcitrant. The absence of external force implies that the technique is less expensive compared to other in-situ techniques.
Engineered in-situ bioremediation
The second approach involves the introduction of certain microorganism to the site of contamination. Genetically Engineered microorganisms used in the in-situ bioremediation accelerate the degradation process by enhancing the physicochemical conditions to encourage the growth of microorganisms.
Bioventing techniques involve controlled stimulation of airflow by delivering oxygen to unsaturated (vadose) zone in order to increase activities of indigenous microbes for bioremediation. In bioventing, amendments are made by adding nutrients and moisture to increase bioremediation. That will achieve microbial transformation of pollutants to a harmless state. This technique has gained popularity among other in-situ bioremediation techniques [25].
This technique combines vacuum-enhanced pumping, soil vapor extraction and bioventing to achieve soil and ground water remediation by indirect providing of oxygen and stimulation of contaminant biodegradation [26]. This technique is planned for products recovery from remediating capillary, light non-aqueous phase liquids (LNAPLs), unsaturated and saturated zones. This technique used to remediate soils which are contaminated with volatile and semi-volatile organic compounds. The method uses a “slurp” that spreads into the free product layer, which pulls up liquids from this layer. The pumping machine transports LNAPLs to the surface by upward movement, where it becomes separated from air and water. In this technique, soil moisture bounds air permeability and declines oxygen transfer rate, which reducing microbial activities. Although this technique is not suitable for low permeable soil remediation, it is cost effective operation procedure due to less amount of ground water, minimizes storage, treatment and disposal costs.
This technique is similar to bioventing in this air is injected into soil subsurface to improve microbial activities which stimulate pollutant removal from polluted sites. However, in bioventing, air is injected in saturated zone, which can help in upward movement of volatile organic compounds to the unsaturated zone to stimulate biodegradation process. The efficiency of biosparging depends on two major factors specifically soil permeability and pollutant biodegradability. In bioventing and soil vapor extraction (SVE), biosparing operation is closely correlated technique known as in-situ air sparging (IAS), which depend on high air-flow rates for volatilization of pollutant, whereas biosparging stimulates biodegradation. Biosparging has been generally used in treating aquifers contaminated with diesel and kerosene.
Phytoremediation is depolluting the contaminated soils. This technique based on plant interactions like physical, chemical, biological, microbiological and biochemical in contaminated sites to diminish the toxic properties of pollutants. Which is depending on pollutant amount and nature, there are several mechanisms such as extraction, degradation, filtration, accumulation, stabilization and volatilization involved in phytoremediation. Pollutants like heavy metals and radionuclides are commonly removed by extraction, transformation and sequestration. Organic pollutants hydrocarbons and chlorinated compounds are mostly removed by degradation, rhizoremediation, stabilization and volatilization, with mineralization being possible when some plants such as willow and alfalfa are used [27, 28].
Some important factors of plant as a phytoremediator include: root system, which may be fibrous or tap depending on the depth of pollutant, above ground biomass, toxicity of pollutant to plant, plant existence and its adaptability to predominant environmental conditions, plant growth rate, site monitoring and above all, time mandatory to achieve the preferred level of cleanliness. In addition, the plant must be resistant to diseases and pests [29]. In phytoremediation removal of pollutant includes uptake, translocation from roots to shoots. Further, translocation and accumulation depends on transpiration and partitioning [30]. However, the process is possible to change, depending on other factors such as nature of contaminant and plant. The mostly plants growing in any polluted site are good phytoremediators. Therefore, the success of any phytoremediation method mainly depends on improving the remediation potentials of native plants growing in polluted sites either by bioaugmentation with endogenous or exogenous plant. One of the major advantages of using plants to remediate polluted site is that some precious metals can bioaccumulate in some plants and recovered after remediation, a process known as phytomining.
This technique is commonly observed as a physical method for remediating contaminated groundwater. However, biological mechanisms are precipitation degradation and sorption of pollutant removal used in PRB method. The substitute terms such as biological PRB, bio-enhanced PRB, passive bioreactive barrier, have been suggested to accommodate the biotechnology and bioremediation aspect of the technique. In general, PRB is an in-situ technique used for remediating heavy metals and chlorinated compounds in groundwater pollution [31, 32].
In-situ bioremediation methods do not required excavation of the contaminated soil.
This method provides volumetric treatment, treating both dissolved and solid contaminants.
The time required to treat sub-surface pollution using accelerated in-situ bioremediation can often be faster than pump and treat processes.
It may be possible to completely transform organic contaminants to innocuous substances like carbon dioxide, water and ethane.
It is a cost effective method because there is minimal site disruption.
Depending on specific site, some contaminants may not be absolutely transformed to harmless products.
If transformation stops at an intermediate compound, the intermediate may be more toxic and/or mobile than parent compound some are recalcitrant contaminants cannot be biodegradable.
When incorrectly applied, injection wells may become blocked by profuse microbial growth due to addition of nutrients, electron donor and electron acceptor.
Heavy metals and organic compounds concentration inhibit activity of indigenous microorganisms.
In-situ bioremediation usually required microorganism’s acclimatization, which may not develop for spills and recalcitrant compounds.
Bioremediation techniques are varied and have demonstrated effective in restoring polluted sites. Microorganisms play fundamental role in bioremediation; consequently, their diversity, abundance and community structure in polluted environments offer insight into the chance of any bioremediation technique providing other environmental factors, which can inhibit microbial activities. Advanced Molecular techniques such as ‘Omics’ includes genomics, proteomics, metabolomics and transcriptomics have contributed towards microbial identification, functions, metabolic and catabolic pathways, with microbial based methods. Nutrient availability, low population or absence of microbes with degradative capabilities, and pollutant bioavailability may delay the achievement of bioremediation. Since bioremediation depends on microbial process, biostimulation and bioaugmentation approaches speed up microbial activities in polluted sites. Biostimulation increase microbial activities by the addition of nutrients to a polluted sample. Microorganisms are abundantly present in different type of environmental condition, it is noticeable that pollutant degrading microbes are naturally present in polluted contaminated sites, their growth and metabolic activities may depends on pollutant type and concentration; later, we can use of agro-industrial wastes, which contains nitrogen, phosphorus and potassium as a nutrient source most polluted sites. Microbial consortium has been reported to degrade pollutants more efficiently than pure isolates [33].
This activity due to metabolic diversities of individual isolates, which potency create from their isolation source, adaptation process, pollutant composition, and synergistic effects, which may lead to complete and rapid degradation of pollutants when such isolates are mixed together [34]. Additional so, both bioaugmentation and biostimulation were effective in removing pollutant such as polyaromatic hydrocarbons (PAHs) from heavily polluted sample compared to non-amended setup (control) [35].
Although bioaugmentation has recognized effective method, it has been shown to increase the degradation of many compounds. If proper biodegrading microorganisms are not present in soil or if microbial populations decreased because of contaminant toxicity, specific microorganisms can be added as “introduced organisms” to improve the current populations and the possibility that the inoculated microorganisms may not survive in the new environment make bioaugmentation a very uncertain method. This process is known as bioaugmentation. Bioremediation technique in which natural or genetically engineered bacteria with unique metabolic profiles are used to treat sewage or contaminated water or soil. The use of alginate, agar, agarose, gelatin, gellan gum and polyurethane as carrier materials solve some of the challenges associated with bioaugmentation [36].
Biosurfactants are chemical equivalents having ecofriendly and biodegradable properties. However, high construction cost and low scalability application of biosurfactants to polluted site are uneconomical. Agro-industrial wastes combination are nutrient sources for development of biosurfactant producers during fermentation process. Application of several bioremediation techniques will help increase remediation efficiency [37].
Enhancing bioremediation ability with organized use of genetically engineered microorganisms (GEM) is a favorable approach. This is due to possibility of engineering a designer biocatalyst target pollutant including recalcitrant compounds by combining a novel and efficient metabolic pathways, widening the substrate range of existing pathways and increasing stability of catabolic activity [38].
However, parallel gene transfer and multiplication of GEM in an environmental application are encouraging approach. Bacterial containment systems, in which any GEM escaping an environment to reconstruct polluted environment.
Further, derivative pathway of genetically engineering microorganisms with a target polluted compound using biological approach could increase bioremediation efficiency. Nanomaterials decline the toxicity of pollutant to microorganisms because nanomaterials having increase surface area and lower activation energy, which reduce time and cost of bioremediation [39].
Bioremediation must be considered as appropriate methods that can applied to all states of matter in the environment
Solids (soils, sediment and sludge)
Liquids (ground water, surface water and industrial waste water
Gases (industrial air emissions)
Sub-surface environments (saturated and vadose zones).
The general approaches to bioremediation are the (i) intrinsic (natural) bioremediation, (ii) biosimulation (environmental modifications, through nutrient application and aeration, and (iii) bioaugmentation (addition of microbes).
The biological community exploited for bioremediation generally consists of the native soil microflora. However, higher plants can also be manipulated to enhance toxicant removal (phytoremediation), especially for remediation of metal contaminated soils.
All bioremediation techniques have its own advantage and disadvantage because it has its own specific applications.
It is a natural process; it takes a little time, as an adequate waste treatment process for contaminated material such as soil. Microbes able to degrade the contaminant, the biodegradative populations become reduced. The treatment products are commonly harmless including cell biomass, water and carbon dioxide.
It needs a very less effort and can commonly carry out on site, regularly without disturbing normal microbial activities. This also eradicates the transport amount of waste off site and the possible threats to human health and the environment.
It is functional in a cost effective process as comparison to other conventional methods that are used for clean-up of toxic hazardous waste regularly for the treatment of oil contaminated sites. It also supports in complete degradation of the pollutants; many of the toxic hazardous compounds can be transformed to less harmful products and disposal of contaminated material.
It does not use any dangerous chemicals. Nutrients especially fertilizers added to make active and fast microbial growth. Because of bioremediation change harmful chemicals into water and harmless gases, the harmful chemicals are completely destroyed.
Simple, less labor intensive and cheap due to their natural role in the environment.
Contaminants are destroyed, not simply transferred to different environmental.
Nonintrusive, possibly allowing for continued site use.
Current way of remediating environment from large contaminates and acts as ecofriendly sustainable opportunities.
It is restricted for biodegradable compounds. Not all compounds are disposed to quick and complete degradation process.
There are particular new products of biodegradation may be more toxic than the initial compounds and persist in environment.
Biological processes are highly specific, ecofriendly which includes the presence of metabolically active microbial populations, suitable environmental growth conditions and availability of nutrients and contaminants.
It is demanding to encourage the process from bench and pilot-scale to large-scale field operations. Contaminants may be present as solids, liquids and gases. It often takes longer than other treatment preferences, such as excavation and removal of soil or incineration.
Research is needed to develop and engineer bioremediation technologies that are appropriate for sites with complex mixtures of contaminants that are not evenly dispersed in the environment.
Bioremediation is limited to those compounds that are biodegradable. This method is susceptible to rapid and complete degradation. Products of biodegradation may be more persistent or toxic than the parent compound in the environment.
Specificity
Biological processes are highly specific. Important site factors mandatory for success include the presence of metabolically capable microbial populations, suitable environmental growth conditions, and appropriate levels of nutrients and contaminants.
Scale up limitation
It is difficult to scale up bioremediation process from batch and pilot scale studies applicable to large scale field operations.
Technological advancement
More research is required to develop modern engineer bioremediation technologies that are suitable for sites with composite combinations of contaminants that are not equally distributed in the environment. It may be present as solids, liquids and gases forms.
Time taking process
Bioremediation takes longer time compare to other treatment options, such as excavation and removal of soil from contaminated site.
Regulatory uncertainty
We are not certain to say that remediation is 100% completed, as there is no accepted definition of clean. Due to that performance evaluation of bioremediation is difficult, and there is no acceptable endpoint for bioremediation treatments.
Biodegradation is very fruitful and attractive option to remediating, cleaning, managing and recovering technique for solving polluted environment through microbial activity. The speed of undesirable waste substances degradation is determined in competition with in biological agents like fungi, bacterial, algae inadequate supply with essential nutrient, uncomfortable external abiotic conditions (aeration, moisture, pH, temperature), and low bioavailability. Bioremediation depending on several factors, which include but not limited to cost, site characteristics, type and concentration of pollutants. The leading step to a successful bioremediation is site description, which helps create the most suitable and promising bioremediation technique (ex-situ or in-situ). Ex-situ bioremediation techniques tend to be more costly due to excavation and transportation from archeological site. However, they can be used to treat wider range of pollutants. In contrast, in-situ techniques have no extra cost for excavation; however, on-site installation cost of equipment, attached with effectively and control the subsurface of polluted site can reduce some ineffective in-situ bioremediation methods. Geological characteristics of polluted sites comprising soil, pollutant type and depth, human habitation site and performance of every bioremediation technique should be integrated in determining the most appropriate and operative bioremediation technique to successfully treatment of polluted sites.
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