Subgroups of clay minerals.
\r\n\tNeutrophil as a white blood cell helps fight off infections and plays a main role in immune response, especially in infectious diseases and phagocytosis. It can also cause inflammation. Neutrophils are a normal cellular component of the blood and also of certain tissues, including spleen, lymph nodes, thymus, and the submucosal areas of the gastrointestinal, respiratory, and genitourinary tracts. This can happen in many different parts of the body, including the esophagus, heart, lungs, blood, and intestines. This cell has the main role in hemostasis, the physiological function of organs, protective role against many diseases such and syndromes. Neutrophil deficiency in function and count of this cell can lead to the beginning and progress of many diseases and problems.
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Venkateswarlu",coverURL:"https://cdn.intechopen.com/books/images_new/371.jpg",editedByType:"Edited by",editors:[{id:"58592",title:"Dr.",name:"Arun",surname:"Shanker",slug:"arun-shanker",fullName:"Arun Shanker"}],productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}}]},chapter:{item:{type:"chapter",id:"72560",title:"Limestone Clays for Ceramic Industry",doi:"10.5772/intechopen.92506",slug:"limestone-clays-for-ceramic-industry",body:'Clays are inorganic, natural, earthy, and fine-grained materials that acquire plasticity when mixed with water [1]. For sedimentologists, a clay is a raw material whose grain size is less than 2 μm. Like clays, in turn, there are rocks made up of clay minerals and may contain other minerals such as quartz, feldspar, mica, calcite, hematite, and organic matter as accessories [2]. A clay, once ground and mixed with water, in addition to presenting excellent workability in the fresh state, after drying, becomes extremely rigid. After burning normally above 800°C, it acquires great resistance [3]. Clays are used worldwide in the ceramic industry, especially in bricks, coatings, and others. However, clays are formed from the weathering of explosion and can be contaminated with several minerals among them or carbonate, which can alter the shape that causes the following burns. Limestone may be present in colloidal form, or coarse particles. However, in all cases it is impossible to separate or calculate this. Some researchers have tried to reduce the size of the variations to improve the chemical changes. According to Barba et al. [4], calcium carbonate and magnesium carbonate are the main constituents of carbonate sedimentary rocks. Anionic carbonate groups are strongly activated units and share oxygen with each other. They are responsible for the properties of these minerals. The most important anhydrous carbonates belong to three isostructural groups: the calcite group, the aragonite group, and the dolomite group. Among these, the minerals most used in the ceramic industry are calcite and dolomite, as they are low-cost raw materials, in addition to having favorable physical and chemical properties and available deposits. Second, Padoa [5] adds that when CaCO3 is small, a decomposition can be complete and the calcium oxide reaches later with other mass components forming calcium silicates and silicon aluminates (wollastonite, anortite, gehlenite etc.) during sintering. Barba et al. [4] mentioned that the raw materials of clay when burned at high temperatures produce crystal phases that influence the properties of ceramic products. Calcite exerts a bleaching action on burnt products when added to a formulated mass of clays (in proportions above 5% and less than 30%) and at the same time decreases its expansion by legislation, as it forms crystalline and liquid phases, including cycles temperature and firing adopted. Calcite and dolomite are the most important representatives of carbonates in the ceramic industry. They are used as main components in the manufacture of ceramic tiles with high water absorption. These coatings include “porous coatings” or “tiles.” These products are designed or used on walls and are not suitable for application on floors, as they have undesirable technical characteristics, such as mechanical resistance, incompatibility with use. According to Amorós [6], properties of parts of a ceramic product are registered by crystalline phases formed based on calcium and magnesium as ghelenite (SiO2⋅Al2O3⋅2CaO) and anortite (2SiO2⋅Al2O3⋅CaO). To achieve these phases, use the dolomite calcium oxide and/or magnesium reaction with a remaining clay structure proven by its thermal decomposition.
The calculation in general can affect the ceramic product in two ways: low percentages (up to 3%) and high temperature (above 1180°C) result in flow agents, that is, materials that contribute to reduce water absorption and increase the resistance of ceramic products. Above 3%, they can act as a foundation at temperatures above 1170°C [7].
In this chapter, we will highlight properties of limestone clays and their application in the ceramic industry.
Clays are hydrated aluminum silicates with crystalline structure arranged in layers, consisting of continuous sheets of SiO4 tetrahedrons, ordered in a hexagonal shape, condensed with octahedral sheets of di and trivalent metal hydroxides, usually below 2 μm. They are materials that in contact with water become plastic, a fundamental characteristic for conformation of ceramic products because it provides mechanical resistance in the pressing, extrusion, or gluing process. Clays are mixtures of various clay minerals such as kaolinite, illite, and montmorillonite, which may or may not contain impurities [3, 8].
The kaolinite with structural formula Al2O3⋅2SiO2⋅2H2O has a dioctahedral structure, which consists of a tetrahedral layer linked by an octahedral layer. Pure kaolinites usually have low plasticity, see Figure 1.
Kaolinite structure. (a) Si▬O tetrahedra on the bottom half of the layer and Al▬O,OH octahedra on the top half. (b) Dioctahedral structure.
Montmorillonites are a set of family of clay minerals, composed of dioctahedral and trioctahedral silicate sheets, see Figure 2(a) and (b). The most outstanding feature of these minerals is their ability to absorb water molecules [8, 9]. It has 80% of exchangeable cations in the galleries and 20% on the lateral surfaces. The modification of montmorillonite clays has aroused scientific and technological interest for providing significant improvements when incorporated into pure polymeric materials and conventional composites. The clay modification process occurs preferably through the ionic exchange of the exchangeable cations of its crystalline structure.
Crystalline structure of a montmorillonite. (a) Montmorillonite structure, composed of Si, Al, and O. (b) Sheets of dioctahedral and trioctahedral silicates.
The basic structural unit of the illites is the same as that of the montmorillonites except that in illites, the silicon atoms in the silica layers are partially replaced by aluminum. Therefore, there are free valences in the boundary layers of the structural units, which are neutralized by K cations, arranged between the overlapping units. The structural scheme of the illites is shown in Figure 3. The K cation is the one that best adapts to the hexagonal meshes of the oxygen planes of the layers of silica tetrahedron and is not displaced by other cations. The water adsorption and cation exchange capacity is due only to the broken connections at the ends of the layers. The average diameter of the illites varies between 0.1 and 0.3 μm. When the replacement of silicon in the tetrahedron layers by aluminum in the illites is small, the connections between the structural units provided by the K cations may be deficient and will allow water to enter. When this occurs, the properties of the illites are close to the properties of montmorillonites [3].
Crystalline structure of an illite. (a) Silicon atoms in the silica layers partially replaced by aluminum in the illites. (b) Structural scheme of illites.
Chlorites are minerals made up of four hydrated aluminum and magnesium silicate layers, containing Fe (II) and Fe (III) as shown in Figure 4.
Crystalline structure of chlorite [9].
The most common clay minerals are interstratified, characteristic of mixtures of clay minerals, classified by subgroup and mineralogical species, see most common classification in Table 1. Clay minerals are divided into several classes. A large majority of clays do not have in just one crystalline phase. Two or more chemical species may be present.
Subgroup | Chemical species | Minerals |
---|---|---|
Kaolin Xn(Y2O5)(OH)4 | Kaolinites | Nacrite (Al2(Si2O5)(OH)4) Dikite (Al2(Si2O5)(OH)4) Livesite (Al2(Si2O5)(OH)4) Halloysite (Al2(Si2O5)(OH)4) |
Talc XB(Y2O5)(OH)2ZmH2O | Montmorillonites | Montmorillonites (Al1,51Fe0,07Mg0,60)(Al0,28Si3,72)O10(OH)2Na0,33 |
Beidellite (Al1,46Fe0,50Mg0,08)(Al0,36Si3,64)O10(OH)2Na0,4 | ||
Nontronite (Fe1,67Mg0,33)(Si4O10)(OH)2Na0,33 and Fe2,22(AlSi3O10)(OH)2Na0,33 | ||
Hectorite (Mg2,67Li0,33)(Si4O10)(F,OH)2Na0,33 | ||
Saponite Mg3(Al0,33Si3,67)O10(OH)2Na0,33 | ||
Illites | Wide variety of minerals | |
Chlorite | Chlorites | Chlorite |
X2n(Y2O5)2(OH)2 | [Mg2(Al,Fe(III))(OH)6][Mg3(AlSi3O10)(OH)2] |
The clays used in the ceramic manufacturing process can be classified into:
Carbonitic clays: they are formed by associations of illitic-chloritic and eventually illitic-kaolinite clay minerals. The amount of calcium carbonate present can be variable. These clays give the dough plasticity. Generally, after burning they have colors ranging from beige to orange [4].
Non-carbonitic clays: they are characterized by the almost total absence of carbonates. The clay minerals present are of the illitic-chloritic type. It has the function of giving plasticity to the dough, and generally after firing they give rise to well-sintered materials.
White plastic clays: the clay matrix is kaolinitic, with little illite. They give plasticity to the dough, and after burning they have a white color.
Kaolinitic clays: clays of low plasticity and normally free of fluxing oxides such as K2O and Na2O, therefore, with refractory characteristics.
According to Mackenzie [10], when a ceramic raw material is subjected to the action of heat, it experiences volumetric variations, usually permanent and irreversible, which can be classified as:
Oxidation of organic matter
Decomposition of compounds containing oxygen, such as sulfates, carbonates, etc.
Dehydroxylation of the clayey mineral
Crystallization by increasing the temperature
Vitreous phase formation
Solid solutions: adjacent crystals of two different materials but of similar structure can react with each other, forming a solid solution.
Kaolinitic clay: the scheme according to Figure 5 shows an endothermic peak between 560 and 590°C referring to the elimination of hydroxyls from the constitution water present in the clays, and an exothermic peak between 980 and 1000°C, due to the formation of mullite, which can be represented by the reactions 1 and 2 [8].
Differential thermal analysis of a kaolinitic clay [10].
Montmorillonite: montmorillonites have water that lodges in the mineral structure, that is, hydration water of adsorbed ions. The elimination of hydroxyl groups occurs at 700°C. At 850°C, a small endothermic peak may occur due to the loss of montmorillonite crystallinity. Illites can present loss of adsorbed water between 100 and 200°C and water loss in the constitution between 550 and 600°C, see Figure 6.
Differential thermal analysis of a montmorillonite clay [10].
Quartz: it appears in clays in colored or colorless round grains, whose percentage ranges from 0 to 60%. For high levels of quartz, the clay is called sandy and has low plasticity [11].
Hematite: iron can be present in the forms of hematite (α-Fe2O3), goethite (α-FeO⋅OH), and lemonade (a mixture of iron oxides and hydroxides of a weakly crystalline nature), or simply as Fe3+ ions in the clay structure. In the illite group, Fe3+ ions can replace Al3+ ions in the octahedral structure [11]. Fe2O3 is formed during sintering under oxidation conditions and from minerals in the clays, giving a reddish color to ceramic materials.
Feldspar: feldspars refer to a group of aluminum silicate minerals. The feldspar contained in the clays is a source of sodium and potassium oxides and plays an important role in ceramic materials with quality of flow agents, temperatures such as sintering temperatures, porosity after firing and facilitating phase formation [6]. The most representative are the orthoclase (KAlSi3O8) and albite (NaAlSi3O8).
Carbonates: calcium or magnesium carbonates can appear as coarse or small grains. If they are presented as large grains (>125 μm), they may not react completely and the resulting oxides may rehydrate causing expansion according to reactions [12, 13].
Ceramic enamels and frits: can be used in matte enamels as a source of CaO to form crystals such as wollastonite, anorthite, gehlenite or in transparent enamels giving shine.
Masses for ceramic coating: as a source of CaO up to the limit of 3%, CaCO3 assists in the formation of the vitreous phase. CaO levels that vary from 8 to 14% favor the formation of crystalline phases such as gehlenite, wollastonite, pseudo wollastonite, and anortite.
Putties for limestone porcelain: calcium carbonates provide the CaO that are used as a flux in limestone porcelain masses.
Ceramic pigments: the calcium carbonate provides calcium oxide, which together with SnO2 produces pink pigments.
Glasses: glasses based on NaOH and CaO use CaCO3 in their composition.
Obtaining settlement mortars: as a plasticizing agent for water retention and aggregate incorporation.
Steel: CaCO3 acts as a flux and pH regulator in water treatment and as lubricant for drawing steel rebars.
Sánchez et al. [14] defined some specification parameters for choosing raw materials for formulations of coating masses, as shown in Table 2 below.
Product | (%) of carbonates | Max. particle size of CaCO3 (μm) | Organic matter (%) | Sulfate content max. (%) | IP (%) |
---|---|---|---|---|---|
Stoned | ≤3 | ≤125 | ≤0.3 | 0.2 | 20–40 |
Porous | ≤40 | ≤125 | ≤0.3 | 0.2 | 20–40 |
Specifications for choosing raw materials.
IP: index of plasticity.
Calcium or magnesium carbonates can appear as coarse or small grains. If they are presented as large grains (>125 μm), they may not react completely, and the resulting oxides may rehydrate causing expansion.
In compositions of ceramic floor covering with low water absorption, CaCO3 acts as a flux until the limit of 3%; above this value, CaCO3 increases porosity and can be accepted up to 40% in porous coatings.
Enrique [15] recommends that the CaCO3 particle size should be less than 125 μm, because particles of larger sizes, the CaO resulting from the dissociation of carbonates when calcined at 900°C, do not react with the SiO2 present in the clays and feldspars that should form the pseudo-wollastonite and wollastonite phases, which can give rise to Ca(OH)2 formed by the hydration of CaO, when the part comes into contact with the humidity of the air, generating problems of expansion by humidity, with consequent cracking.
The ceramic tile and brick industry have grown enormously in recent years in Brazil. The clays must have sufficient plasticity to provide mechanical resistance when forming by pressing, in order to guarantee the integrity of the piece in the path between the press and the oven. The feldspar contained in the clays are sources of sodium and potassium oxides, acting as fluxes at temperatures above 800°C for bricks and above 1100°C for ceramic tiles, which facilitates the formation of a vitreous phase and reduces porosity [16, 17].
Quartz is mixed with clay during geological formation. If it is present in a smaller proportion, it helps in the formation of the vitreous phase, in the degassing of organic matter and water. However, large proportions of quartz lead to a drastic reduction in mechanical strength after firing [18]. Iron oxide is present in ceramic raw materials in the form of hematite or goethite, giving the finished product a red color.
Calcite, which appears in most clays used in the production process of ceramic tiles of type BIIb, is a mineral that needs special care in its use due to its high loss to fire. When present in a proportion equal to or less than 3%, this mineral acts as a flux. However, in higher proportions, calcite can cause an increase in the final porosity of the product. In addition, the size of the calcite particle for processing ceramics must be less than 125 μm. For larger sizes, it is observed that the CaO resulting from the dissociation of carbonates can hydrate after burning, promoting variations in the dimension of the piece. Therefore, the use of limestone clays is a challenge, requiring care in processing and control in the formulation and burning of coatings. To ensure the correct sintering of the product, proper grinding and pressing of the raw material are necessary, in addition to efficient, fast burning with the lowest possible energy consumption.
Table 3 shows the chemical compositions of a typical Brazilian limestone clay used in ceramics [19]. The chemical compositions of the raw materials were determined by X-ray fluorescence spectroscopy by wavelength dispersion (WDFRX), in a Bruker S8 Tiger equipment, in which the percentages of constituent oxides were estimated by the method semi-quantitatively. For these measurements, samples with a mass of 10.0 g were pressed as discs with 40.0 mm diameter and 4.0 mm thickness. During measurements, the samples were kept in a vacuum of 10−6 bar. A mixture of P-10 (90% argon and 10% methane) was used in the proportional counter.
Oxide (%) | C1 | C2 | C3 | C4 |
---|---|---|---|---|
SiO2 | 63.0 | 52.1 | 50.2 | 45.3 |
Al2O3 | 16.7 | 18.6 | 15.5 | 14.1 |
Fe2O3 | 4.7 | 6.8 | 6.2 | 7.1 |
CaO | 0.9 | 2.1 | 7.2 | 12.7 |
K2O | 3.8 | 4.7 | 3.2 | 3.2 |
Na2O | 0.6 | 0.4 | 0.5 | 0.7 |
MgO | 1.5 | 2.3 | 2.2 | 2.3 |
TiO2 | 0.6 | 0.8 | 0.7 | 0.8 |
L.O.I | 8.2 | 12.1 | 14.3 | 13.8 |
The results show that all clays are composed mainly of SiO2 and Al2O3. These elements are associated with clay minerals, quartz, and feldspar structures [17]. The highest amount of SiO2 was determined for sample C1. This component is important for the manufacture of ceramic tiles, as it improves workability and favors compaction. However, SiO2 can also cause low mechanical strength of sintered ceramic bodies, in addition to reducing shrinkage during firing.
The amount of Fe2O3 detected in the samples was between 4.7 and 7.1%. These values are acceptable for use in ceramic tiles, such as bricks and tiles, this element being responsible for the reddish color of the sintered pieces as well as being a powerful flux [20]. The high content of calcium oxide in C4 (12%) and C3 (7%) stands out, characterizing these clays as limestone [21]. C4 clay was previously studied in Alcântara [16], which reports the formation of stains on the ceramic bodies produced with this material, after sintering at 1120°C. This behavior was associated with a high content of CaO, estimated at 10%, which during the burning phase, the dissociation of CaCO3, promotes a high mass loss. C4 (13%) generates many pores, reducing water absorption and resistance of the final product. Thus, the higher the CaO content, the higher the CaCO3 content and in addition, the higher the mass loss.
Analyzing the levels of alkaline oxides, it is observed that the sample C2 has the highest concentration of K2O, while the concentration of Na2O is approximately the same in the four samples studied. Alkaline and alkaline earth compounds have a melting effect, which facilitates the formation of liquid phase and linear shrinkage during burning [13].
Table 4 was arranged according to the increasing amount of CaO present in the clays. Note that C1 and C2 have CaO content below 3%. According to Enrique [15], CaO acts as a flux until the limit of 3% in masses of ceramic coating. The percentage of alkali oxides (Na2O and K2O), also presented in Table 3, is another major factor for the densification process, due to the great tendency of liquid phase formation during burning. Considering the sum of the percentages of CaO and alkali oxides in samples C3 and C2, it can be concluded that C2 has a higher proportion of fluxing oxides, suggesting that this sample is the most promising. On the other hand, clays with a high limestone content, such as C3 and C4, tend to have greater porosity and less mechanical resistance after firing. Additionally, these two raw materials have lower alkaline oxide ratios than those observed for C3 and C2.
Clay | CaO (%) | Na2O + K2O (%) |
---|---|---|
C1 | 0.9 | 4.4 |
C2 | 2.1 | 5.1 |
C3 | 7.2 | 3.7 |
C4 | 12.7 | 3.9 |
The X-ray diffraction patterns of the clays are shown in Figure 7 and correlate positively with the results observed by X-ray fluorescence. The X-ray diffractometry (XRD) technique was used to determine the crystalline phases. The samples were dried in an oven at 110 °C for 24 h, ground, and passed through a 150-μm mesh sieve. The diffraction patterns were obtained in a Rigaku D-MAX 100 equipment, using Cu Kα1 radiation (λ = 1.5418 Å). All measurements were carried out in the continuous scanning mode with speed of 1°/min, in the range of 5 to 65° and in the range of 2 to 15° in samples saturated with ethylene glycol for 1 h to identify montmorillonite by displacing the diffraction peaks at smaller angles compared to dry sample testing. The crystalline phases were identified through Match! (Phase Identification by Powder Diffraction) in the demo version, according to the ICSD (Inorganic Crystal Structure Database).
X-ray diffraction patterns of the clays [19].
The main phases identified were quartz, kaolinite, muscovite, montmorillonite, calcite, feldspar, and hematite. Minerals from kaolinite and montmorillonite clay were identified in all analyzed clays. According to Celik [20], these clay minerals provide the necessary plasticity to guarantee conformation through the pressing process. The percentage of each crystalline phase present in the samples was estimated from the relative intensity of the main peaks in each phase. The values are shown in Table 5. The percentage of carbonates increases from 0.9% in C1 to 12.4% in C4.
Minerals (%) | C1 | C2 | C3 | C4 |
---|---|---|---|---|
Quartz | 55.7 | 51.8 | 65.1 | 57.1 |
Kaolinite | 6.3 | 10.7 | 7.4 | 5.5 |
Muscovite | 11.8 | 14.0 | 11.2 | 12.1 |
Montmorillonite | 5.6 | 4.9 | 4.6 | 6.7 |
Calcite | 8.6 | 2.8 | 1.1 | 13.7 |
Feldspar | 6.3 | 9.9 | 6.2 | 3.2 |
Hematite | 5.7 | 5.9 | 4.4 | 1.7 |
Mineralogical compositions of clays determined by XRD.
To verify the dimensional changes of expansion and thermal retraction of the samples, dilatometry tests were performed on a Netzsch dilatometer, model DIL 402PC, under synthetic air flow at 130 ml/min. For these analyses, the samples were compacted in a cylindrical shape, 12.0 mm in length and 6.0 mm in diameter. Under a constant heating rate of 10°C/min, the length of the compacted body is measured as a function of time and temperature, which varied from room temperature to 1150°C.
In Figure 8 we can observe a slight expansion in all curves up to approximately 850°C, and at 573°C, the expansion was more pronounced due to the transformation of α quartz to β [22, 23], except for C2, which presents a lower percentage of free quartz. From 573°C, there was a gradual reduction in the expansion rate, occurring or starting with sintering, followed by an exponential retraction [22].
Dilatometric curves of clays at a heating rate of 10°C/min [19].
The results shown in Table 5 with the percentages of CaO, Na2O, and K2O recommended by XRF measurements point out that sample C2 has a greater amount of funds (calcium carbonate up to a limit of 3% and alkaline oxides), or what is known as a greater linear shrinkage. Despite its advantages over the other samples, the C2 clay underwent deformation during firing up to 1150°C. This effect, known as pyroplastic deformation, may be due to the large proportion of funds in the sample, a high content of Fe2O3, and, even, the amount of organic matter [24]. One of the ways to control deformation during firing is to adjust the thermal cycle through the dilatometric curves, so that the plate remains within the required standards [25].
Clays containing limestone when subjected to burning, CaCO3 after heating, in the temperature range between 850 and 920°C, form CaO and release CO2. An intense endothermic peak of approximately 35–44% of the mass loss can be observed in differential thermal analysis. In ternary diagrams, it is observed that there is a eutectic point (above 1170°C), which reduces the dimensional stability in ceramic products, which can melt quickly (Figure 9).
Ternary diagram of CaO, SiO2, and Al2O3.
Clays when mixed with limestone can behave differently, as shown by Sánchez [25]. Figure 10 shows a standard clay with 5 and 10% of incorporated limestone. It was observed that as the limestone and temperature increase, respectively, the dimensional instability increases. In other words, the retraction increases constantly, when it undergoes an exponential increase, reaching the melting point.
Ceramic coating mass with incorporated calcite waste.
This phenomenon can be explained as follows: when exhibiting CaO up to the limit of 3%, this, associated with SiO2 and Al2O3 present in clays and feldspars, helps in the formation of eutectic systems at 1170°C, with consequent formation of liquid phase and contributing to obtain the desired mechanical strength and porosity. When introduced in percentages above 4%, CaCO3 levels are increased, and the composition moves from the eutectic line, forming crystalline phases such as CaSiO3 (pseudo-wollastonite) and 2CaO⋅Al2O3⋅SiO2 (gehlenite). So, a larger number of pores is left by the eliminated CO2. In this way, the porosity of the final product is increased, as shown in Figure 11. In Figure 12 is shown a photo of a clay mass with 10% calibration in which the porosity exerted can be observed.
Firing curve of a calcite clay.
Scanning electron microscopy of a ceramic with 10% of CaO.
Limestone is a contaminant for clay that above 125 μm can cause expansion and consequently cracks.
Rapid tests that mix clay with HCl can promote effervescence due to the release of CO2 and contribute to decrease the amount of limestone.
In the ceramic industry, wet grinding of components is carried out in ball mills and grinding will be more efficient if the sieves are 150 to 325 μm. In ceramic mass formulations, the amount of CaO up to 3% contributes to the formation of the vitreous phase, however, between 8 and 14%, it favors the formation of crystalline phases, reducing the absorption of water and increasing the mechanical resistance.
The aquatic environment is highly complex and diverse, comprising various types of ecosystems that are dynamic products of complex interactions between biological and abiotic components. Changes in physical properties and ecosystems may affect the balance of life forms present there [1, 2].
\nIn recent decades, these ecosystems have been significantly altered due to multiple environmental impacts from the release of large amounts of effluent without adequate prior treatment, resulting in the scarcity of existing natural resources [3, 4]. Among the main pollutants that generate negative impacts on life forms are heavy metals. The presence of these contaminants may cause changes in the structure and function of microbial communities [5], which can develop various resistance mechanisms that enable their survival [6]. In addition, heavy metal resistance may contribute to the evolution of resistance genes to different types of antimicrobials due to increased selective pressure in the environment [7].
\nAdaptability as well as metabolic and physiological differences are essential characteristics for microorganisms to remain in these locations. One of the adaptive mechanisms present in bacteria that has been frequently investigated is biofilm formation [8]. Biofilms are structures composed mainly of microbial cells and a matrix formed by a cluster of extracellular polymeric substances (EPSs) [9]. Biofilm-grown cells have some distinct properties from planktonic cells, one of which is increased resistance to antimicrobials and heavy metals [10]. In this review, we propose to report the latest findings on the survival strategies of microorganisms in impacted aquatic environments, more precisely on the influence of heavy metals on biofilm formation.
\nWater is an indispensable natural resource for the survival of man and other living beings [11, 12]. According to Raucci and Polette [13], 97% of the planet’s water is found in the oceans, and of the remaining 3%, only 0.3% is available for human consumption and is stored in springs, lakes, rivers, and groundwater.
\nAccording to the United Nations (UN), access to water supply and sanitation is a human right and vital to the dignity and health of all people. However, there are still about 1.1 billion people without access to clean water and 2.4 billion people without access to basic sanitation services [14].
\nThe decline in water quality has become one of the most serious problems worldwide, a fact that has been intensified by the increase in population and the absence of public policies aimed at the preservation of water resources. According to the World Health Organization—WHO [15], approximately half of the world’s developing population will be affected by diseases that are directly related to poor-quality water and/or lack of adequate or even no sanitation.
\nContamination of natural waters represents one of the main risks to public health, a fact that is directly related to the discharge of untreated domestic, hospital, and industrial effluents, which cause contamination of aquatic bodies by pathogenic microorganisms such as bacteria, viruses, protozoa, and helminth eggs [16].
\nAmong the bacteria can be highlighted those belonging to the Enterobacteriaceae family, represented by species Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Enterobacter cloacae and Providencia rettgeri. Most of these species are commonly found in the intestinal tract of humans and animals, and their presence in aquatic environments indicates fecal contamination [4, 17].
\nAnother problem found in the aquatic environment is the contamination by resistant bacteria from humans and animals exposed to antimicrobials [18, 19], as well as the disposal of antimicrobial waste from domestic and hospital effluents. Water is not only a means of spreading resistant microorganisms, but also the pathway through which resistance genes are introduced into the ecosystem, altering the environmental microbiota [20].
\nStudies have shown bacterial resistance in various aquatic environments including rivers and coastal areas, domestic sewage, hospital sewage, sediment, surface water, lakes, oceans, and drinking water [4, 21–24].
\nAmong the main pollutants found in this environment, we highlight the heavy metals that when introduced into the environment can cause changes in the structure and function of microbial communities [25]. Aquatic systems may be introduced as a result of natural processes such as weathering, erosion, and volcanic eruptions [26]. However, in recent decades, the increase in urbanization and industrialization has contributed to the large increase of these environmental contaminants worldwide [27].
\nThus, microorganisms have been developing various resistance mechanisms that allow their survival [6]. Among the various mechanisms, intra and extra-cellular, are bioaccumulation [28], biosorption [29], biomineralization and precipitation [30, 31], oxidation and enzymatic reduction of the metal to the less toxic form [32], production of siderophores [33], and biofilm formation [34]. Figure 1 shows an impacted aquatic environment and a survival strategy for the microorganisms present there.
\nImpacted aquatic environment and survival strategy of the present microorganisms.
The term “heavy metals” is used to identify a group of chemical elements that have atomic density greater than 5 g cm−3 or have atomic number greater than 20 [35]. Some of these elements, such as sodium (Na), magnesium (Mg), potassium (K), calcium (Ca), zinc (Zn), and copper (Cu), are essential microelements for various life forms, as they are necessary for the functioning of some metabolic pathways [36]. However, the excess or lack of these elements can lead to disturbances in organisms, and in extreme cases, even death [37]. Other elements such as mercury (Hg), lead (Pb), cadmium (Cd), and arsenic (As) are highly toxic even when present in low concentrations, and account for most health problems due to environmental pollution [38].
\nHeavy metals participate in the global ecobiological cycle, derived from numerous sources and are dynamically transported through the atmosphere, soil, and water; also, because they are not biodegradable, they can remain in the environment for long periods [39].
\nAmong the various metals, mercury, cadmium, and lead stand out for being associated with contamination of the aquatic environment, which can cause problems of poisoning to man and other organisms. These elements are capable of reacting with molecules and ligands present in cell membranes, conferring them with the properties of bioaccumulation, food chain biomagnification, persistence in the environment, and metabolic disturbances of living beings [40].
\nBiofilm is a porous and complex structure formed by one or more species of microorganisms, organized in several layers irreversibly adhered to a biotic or abiotic surface and enclosed in a matrix composed of extracellular polymeric substances (EPS) [9].
\nThey are formed dynamically and gradually, involving several stages. The first is reversible bacterial adhesion that can occur on biotic surfaces mediated by molecular interactions or abiotic surfaces through physicochemical interactions. The second is irreversible adhesion, where the adhesion process is consolidated through the production of EPS. After the establishment and maturation of the protective matrix in the irreversible phase, the cycle ends with the rupture of the biofilm and the release of bacterial cells (Figure 1) [9, 41].
\nBacteria in the form of free (planktonic) cells are not often found in nature; most of them live in communities or attached to various biotic or abiotic surfaces, such as clinical and industrial equipment. Several factors may contribute to bacterial adhesion such as flagella, fimbriae, adhesin, and polymers, as well as adhesion forces such as electrostatic and hydrophobic attraction, van der Waals interactions, hydrogen bridges, and covalent bond [10].
\nBiofilm formation is an effective strategy for microbial survival and persistence under stress conditions, such as in the presence of antimicrobials and heavy metals [42]. The biofilm structure may be associated with a protective mechanism that allows the bacteria to survive and persist in environments with high metal concentrations [43]. Studies have shown that subinhibitory heavy metal concentrations can induce biofilm formation [44, 45], like lead [46], cadmium [47], and nickel [48] among others.
\nGiovanella et al. [46] evidenced the increase in formation by an isolate of Pseudomonas sp. in the presence of mercury (Hg2+). Similarly, Araújo et al. [49] verified an increase in biofilm formation in Klebsiella pneumoniae isolates obtained from an impacted urban stream. However, other studies show that depending on the metal and its concentration, biofilm formation may be reduced [50, 51]. These differences may be related to the fact that the effects of metals depend on their concentration and speciation [47, 51, 52], growth conditions, and especially the bacterial isolate that is being exposed [53, 54].
\nRecent studies have shown that metals can affect various stages in biofilm formation and development [55]. Metals can impact cell surface adhesion and/or cell-to-cell aggregation process, promoting biofilm formation and, consequently, its resistance. Harrison et al. [56] verified that the increase in cadmium concentration induces cell adhesion and biofilm formation in Rhizobium alamii YAS34. Subinhibitory concentrations of manganese (Mn) and zinc (Zn) affected cell aggregation in Xylella fastidiosa isolates. Mn increased the process of biofilm formation in this bacterium, while Zn impaired this process probably by reducing cell adhesion on the surface [50, 57]. Perrin et al. [48] observed that some isolates of Escherichia coli K-12 formed biofilm in response to subinhibitory nickel (Ni) concentrations and that cells embedded in the biofilm were less affected by metal exposure than planktonic cells. These studies show that bacterial cells exposed to metals generally respond by inducing adhesion processes, and consequently, biofilm formation and maintenance [55].
\nIn addition to changes in cell adhesion, exposure to heavy metals may cause structural changes in the biofilm extracellular polymeric substance (EPS) matrix. Araújo et al. [49] verified by scanning microscopy, the increase of EPS in K. pneumoniae biofilms formed when exposed to subinhibitory mercury concentrations (Hg2+). Sheng et al. [58] also demonstrated that heavy metals stimulate EPS production in Rhodopseudomonas acidophila. Schue et al. [59] observed in R. alamii isolates the formation of a more condensed biofilm in the presence of subinhibitory concentrations of Cd when compared to isolates not exposed to this metal. The increase of extracellular matrix in Thiomonas sp. subinhibitory concentrations of arsenic (III) possibly contributed to biofilm integrity and physiological heterogeneity of immobilized cell subpopulations [60].
\nIn stabilized biofilm, the presence of metals impacts cells via passive processes by the influence of gene expression, resulting in mechanisms of resistance or tolerance to these pollutants [55]. Extracellular polymeric matrix (EPS) acts as a barrier to toxic metals, which can be sequestered, immobilized, mineralized, and precipitated, diminishing their effect on bacteria [61]. In Pseudomonas putida ATCC 33015, sugars present in the biofilm matrix exposed to chromium (Cr) probably facilitated the immobilization process of this metal [62]. The biomineralization process was described in Cupriavidus metallidurans CH34, which was able to form gold (Au) nanoparticles in biofilm through the reduction and precipitation mechanism of the toxic gold complex (Au III) [63].
\nEnvironmental contamination by heavy metals has been increasing in recent years, due to various anthropogenic activities. Heavy metals, because they are not biodegradable, have a tendency for biomagnification and bioaccumulation and are extremely toxic to various biological functions, causing serious impacts on the environment and human health [64].
\nMicroorganisms present in contaminated environments have developed different resistance mechanisms to adapt to stress caused by heavy metals. The ability to survive under these extreme conditions depends on acquired biochemical and physiological attributes, as well as genetic adaptations [65].
\nSeveral studies suggest that metal contamination in the natural environment may play an important role in maintaining and proliferating antimicrobial resistance (Table 1) [67–69]. In the environment, selective pressure exerted by metals may select resistant isolates similar to antibiotics, since both resistance genes are often located on the same moving elements [70, 71].
\nResistance mechanisms | \nHeavy metals | \nAntibiotics | \nReferences | \n
---|---|---|---|
Reduction in permeability | \nAs, Cu, Zn, Mn, Co, Ag | \nCip, Tet, Cholr, β-lactâmicos | \n[32, 74] | \n
Drug and metal alteration | \nAs, Hg | \nβ-lactâmicos, Chlor | \n[75, 76] | \n
Drug and metal outflow | \nCu, Co, Zn, Cd, Ni, As | \nTet, Chlor, β-lactâmicos | \n[77, 78] | \n
Cell signaling change | \nHg, Zn, Cu | \nCip, β-lactâmicos, Trim, Rif | \n[79, 80] | \n
Examples of characteristics and negative effects on metal and antibiotic resistance mechanisms.
\nAbbreviations: Cholr, chloramphenicol; Cip, ciprofloxacin; Rif, rifampicin; Tet, tetracycline; Trim, trimetropim. Adapted from Baker-Austin et al. [66].
Bacteria develop some mechanisms to neutralize mercury toxicity, the most common being enzymatic reduction of the highly toxic mercuric ion (Hg2+) to the volatile and less toxic elemental mercury (Hg0). This reduction is catalyzed by the cytosolic mercury reductase (MerA) enzyme encoded by a gene belonging to the operon mer. Studies have shown the frequent association between operon mer and antimicrobial resistance [66, 72]. Péres-Valdespino et al. [73] demonstrated that several clinical isolates of Aeromonas sp. that presented the merA gene were resistant to different antibiotics such as tetracycline, trimethoprim, nalixidic acid, and streptomycin. Araújo et al. [49] verified, when comparing isolates of K. pneumoniae, that the isolate that presented the merA gene was resistant to the highest number of antimicrobials and presented the minimum inhibitory concentration (MIC) value up to four times higher than the others, suggesting a co-resistance mechanism for mercury and antimicrobials tested.
\nMartins et al. [81] observed that isolates of P. aeruginosa, obtained from a contaminated river in southeastern Brazil, had a conjugative plasmid with co-resistance to tetracycline and copper, reinforcing that resistance to antibiotics may be induced by selective pressure of heavy metals in the environment. Caille et al. [82] demonstrated that in P. aeruginosa, copper can induce imipenem resistance by the CopR-CopS two-component regulatory system mechanism. Ghosh et al. [83] verified resistance to ampicillin, arsenic, chromium, cadmium, and mercury in Salmonella abortus equi isolates and observed that after removal of the plasmids, isolates became sensitive to these compounds.
\nIn order to corroborate the evidence of co-resistance of metals and antibiotics, some studies compared the resistance profiles of bacteria collected in contaminated and uncontaminated environments. Rasmussen and Sørensen [84] demonstrated an increase in the occurrence of conjugative plasmids at contaminated sites and found that the mercury and tetracycline resistance genes were located on the same plasmid. Mcarthur and Tuckfield [85] examined metal and antibiotic resistance profiles in contaminated and uncontaminated stream sediments and found that isolates obtained from the contaminated sediment were more resistant to kanamycin and streptomycin than the others.
\nThus, not only the indiscriminate use of antibiotics but also environmental contamination by heavy metals can pose risks and harm to human health, as resistance genes can be transferred horizontally from environmental microorganisms to human diners [66].
\nIncreased urbanization and industrialization have contributed to heavy metal contamination in aquatic ecosystems, modifying the structure and function of microbial communities. The ability of microorganisms to survive under stress conditions, such as in the presence of heavy metals, depends on structural and biochemical attributes, as well as physiological and/or genetic adaptations. The studies cited demonstrated that the presence of heavy metals influences at different stages of biofilm formation. Additionally, the correlation between resistance to metals and antimicrobials was demonstrated, showing the environmental impact that these contaminants can cause in aquatic environments.
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I am also a member of the team in charge for the supervision of Ph.D. students in the fields of development of silicon based planar waveguide sensor devices, study of inelastic electron tunnelling in planar tunnelling nanostructures for sensing applications and development of organotellurium(IV) compounds for semiconductor applications. I am a specialist in data analysis techniques and nanosurface structure. 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After obtaining a Master's degree in Mechanical Engineering, he continued his PhD studies in Robotics at the Vienna University of Technology. Here he worked as a robotic researcher with the university's Intelligent Manufacturing Systems Group as well as a guest researcher at various European universities, including the Swiss Federal Institute of Technology Lausanne (EPFL). During this time he published more than 20 scientific papers, gave presentations, served as a reviewer for major robotic journals and conferences and most importantly he co-founded and built the International Journal of Advanced Robotic Systems- world's first Open Access journal in the field of robotics. Starting this journal was a pivotal point in his career, since it was a pathway to founding IntechOpen - Open Access publisher focused on addressing academic researchers needs. Alex is a personification of IntechOpen key values being trusted, open and entrepreneurial. Today his focus is on defining the growth and development strategy for the company.",institutionString:null,institution:{name:"TU Wien",country:{name:"Austria"}}},{id:"19816",title:"Prof.",name:"Alexander",middleName:null,surname:"Kokorin",slug:"alexander-kokorin",fullName:"Alexander Kokorin",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/19816/images/1607_n.jpg",biography:"Alexander I. Kokorin: born: 1947, Moscow; DSc., PhD; Principal Research Fellow (Research Professor) of Department of Kinetics and Catalysis, N. Semenov Institute of Chemical Physics, Russian Academy of Sciences, Moscow.\r\nArea of research interests: physical chemistry of complex-organized molecular and nanosized systems, including polymer-metal complexes; the surface of doped oxide semiconductors. 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