\r\n\tNotably, the book encourages academic scholars and researchers to contribute to the modern concepts of CSR. Fundamentally, it speaks for well-developed literature for entrepreneurs and managers, thus assisting them in the decision-making process. \r\n\tFurthermore, this book is of great value to policymakers, practitioners, and corporations, thus contributing to various disciplines (e.g., social science and management). \r\n\tThese proposed themes encourage future researchers and professionals to share their ideas, concepts and work concerning these subject domains. All these suggested topics had recommended under the rubrics of CSR. Perhaps, all the professionals, researchers, and scholars are welcome to submit their piece of work, in particular to the suggested topics. \r\n\tIndeed, the recommended topics include the following but are not limited to these only. \r\n\t• Corporate Governance and Sustainability \r\n\t• Green Innovation and CSR \r\n\t• Social Entrepreneurship \r\n\t• Green Economy and Social and Environmental Sustainability \r\n\t• Sustainable Development and Industrialization
",isbn:"978-1-80356-165-3",printIsbn:"978-1-80356-164-6",pdfIsbn:"978-1-80356-166-0",doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!1,isSalesforceBook:!1,hash:"e3be182f32c4d9b8e44e95e86ee1366b",bookSignature:"Dr. Muddassar Sarfraz",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/11602.jpg",keywords:"Sustainability, Stakeholders, Corporate Citizenship, Sustainable Development, Decision-making Process, CSR, Organizational Performance, Financial Performance, Corporate Reputation, Environmental Performance, Environmental Strategy, Green Innovation",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:null,numberOfDimensionsCitations:null,numberOfTotalCitations:null,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"February 2nd 2022",dateEndSecondStepPublish:"March 2nd 2022",dateEndThirdStepPublish:"May 1st 2022",dateEndFourthStepPublish:"July 20th 2022",dateEndFifthStepPublish:"September 18th 2022",remainingDaysToSecondStep:"3 months",secondStepPassed:!0,currentStepOfPublishingProcess:4,editedByType:null,kuFlag:!1,biosketch:"A pioneering researcher in Business Management and Sustainability, Associate Editor of Frontiers in Psychology Journal, and published several research articles.",coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"260655",title:"Dr.",name:"Muddassar",middleName:null,surname:"Sarfraz",slug:"muddassar-sarfraz",fullName:"Muddassar Sarfraz",profilePictureURL:"https://mts.intechopen.com/storage/users/260655/images/system/260655.jpeg",biography:"Dr. Muddassar Sarfraz works as an assistant professor at Wuxi University, China. He completed a postdoctoral fellowship in Business Management at the Business School of Hohai University, China. He has published numerous papers in foreign authoritative journals and academic conferences at home and abroad. He is senior editor of Cogent Business & Management, associate editor of Frontiers in Psychology, Energies, and Future Business Journal, and guest editor of Frontiers in Environmental Sciences and INQUIRY. He is a member of the British Academy of Management, Chinese Economists Society (USA), World Economic Association (UK), and the American Economic Association, and an ambassador of the MBA program at Chongqing University, China. His research focuses on corporate social responsibility, risk management, strategic management, and business management.",institutionString:"Zhejiang Shuren University",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"0",totalChapterViews:"0",totalEditedBooks:"2",institution:{name:"Zhejiang Shuren University",institutionURL:null,country:{name:"China"}}}],coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"7",title:"Business, Management and Economics",slug:"business-management-and-economics"}],chapters:null,productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},personalPublishingAssistant:{id:"429339",firstName:"Jelena",lastName:"Vrdoljak",middleName:null,title:"Ms.",imageUrl:"https://mts.intechopen.com/storage/users/429339/images/20012_n.jpg",email:"jelena.v@intechopen.com",biography:"As an Author Service Manager, my responsibilities include monitoring and facilitating all publishing activities for authors and editors. From chapter submission and review to approval and revision, copyediting and design, until final publication, I work closely with authors and editors to ensure a simple and easy publishing process. I maintain constant and effective communication with authors, editors and reviewers, which allows for a level of personal support that enables contributors to fully commit and concentrate on the chapters they are writing, editing, or reviewing. I assist authors in the preparation of their full chapter submissions and track important deadlines and ensure they are met. I help to coordinate internal processes such as linguistic review, and monitor the technical aspects of the process. As an ASM I am also involved in the acquisition of editors. 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1. Introduction
Crystallization, commonly defined as a process of formation of a crystalline solid from a supersaturated solution, melt or vapor phase, is an old technique widely used in laboratory and in industrial processes to separate and purify substances. In various modern industries, crystalline forms with a certain habit, size and structure, constitute the basic materials for the production of highly sophisticated materials [1, 2]. Integrated circuits as well as piezoelectric and optical materials are just a few examples of devices whose properties are dependent on the crystal structure. Also, in organic chemistry, molecular crystals with determined characteristics are now-a-days of utmost importance for the production of pharmaceuticals, dyestuffs, pigments, foodstuffs, chemicals, cosmetics, etc. For all these reasons, crystal growth has become an important and attractive research field.
Crystallization from solution is one of the preferred methods to obtain a crystal since it can be carried out under different experimental conditions and provides a wide variety of products. In fact, it can occur by lowering the temperature of a supersaturated solution, partial evaporation of the solvent, precipitation by adding an anti-solvent or vapor diffusion of a gas into the solution. Furthermore, solvents with different properties can be used. Such a diversity of experimental conditions has great influence in the output, i.e., in the type of crystalline phase that can be obtained. Therefore, any aprioristic selection of the conditions leading to a desired final product is a fundamental challenge but it is also an almost unfeasible task [3-5]. On respect to the harvest, crystallization is, in a certain extension, a trial-and-error operation.
The various steps of crystallization from solvents are summarized in the following scheme:
Scheme 1.
Main crystallization pathways from solution
The competition between the solute-solvent and solute-solute interactions to form the nuclei, and between the specific aggregation forces and packing in order to minimize repulsive interactions, determine the structure of the new crystalline phase. During this process the molecular conformation can change drastically in an unpredicted way. Various computational approaches have been employed to predict the crystalline structure (unit cell, space group and atomic coordinates) formed by a given molecule [6-8], most of them relying on the assumption that the most probable forms are those with lowest lattice energy. Despite the recent success of these methods in the prediction of crystalline structures of small rigid molecules [9], their applicability to high size flexible molecules is not yet satisfactory [10, 11]. The main drawbacks lie in the difficultly of describing accurately the high complexity of inter and intramolecular interactions (e.g. hydrogen bonds and van der Waals interactions) and of selecting, among the energetically feasible polymorphs, that or those that really exist [3]. In truth, crystal prediction is still a very hard task so far.
Nevertheless, computational calculation can be a powerful tool to get information about the different steps of crystallization. The main goal of the present chapter is included in this research field. Quantum-chemical calculations or molecular dynamics simulations, adapted to the molecular complexity can be used for this purpose. In molecular terms, a compound with a certain conformational flexibility exists in a supersaturated aqueous solution as a mixture of conformers with different populations. The knowledge of the conformational equilibrium in this medium is a way of identifying the conformers that are likely to be involved in the early stages of nucleation and to interpret the mechanism of the crystallization process. In this chapter, these topics are addressed by presenting the results obtained for erythritol and L-threitol, two isomeric alditols with four carbon atoms, and glutamic acid. These compounds were chosen because of the role played by their conformational features on crystallization. Furthermore, the two polyols exhibit a different crystallization behavior despite being chemically similar. The energies of the different conformers were obtained through theoretical calculations at the DFT level of theory [12, 13] using the popular B3LYP (Becke, 3-parameter, Lee-Yang-Parr) exchange-correlation functional [14-16] combined with the aug-cc-pVDZ [17, 18] or 6-311++G(d,p) [19] basis set. Solvent effects have been accounted by using the well-established conductor-like polarizable continuum model (CPCM) [20-22].
As crystallization proceeds, the self-association of the solute molecules gives rise to molecular aggregates which play an important role in the resulting crystalline structure [23-25]. Data from the application of molecular dynamics simulations in the study of aqueous solutions of erythitol and L-threitol near the saturation concentration are reported in an attempt to give an insight into the type of supramolecular species formed in solution, and hence a step forward to elucidate about their crystallization behavior.
As stated before, depending on the experimental conditions and intrinsic molecular features of the molecules, different crystalline phases or polymorphs can be obtained by crystallization. The polymorphs differ from one another in the arrangement (packing) and/or conformation of the molecules in the crystal lattice [26-29]. They are commonly called packing polymorphs in the first case and conformational polymorphs in the second one. Polymorphism has significant implications in a wide range of areas and for this reason it has been matter of extensive investigation. Some typical examples are here reported with the objective of pointing out the diversity of structures that can obtained by crystallization.
2. Nucleation and crystal growth
Fluctuations of the order of a solution, for example density, are accompanied by the formation of solute molecular clusters. The driving force to bring the molecules close together is given by the following chemical potential difference:
μ−μ∗=kTlnCC*E1
In this equation, μ and μ* are the chemical potentials of the solute at the actual (C) and saturation (C*) concentrations, respectively. The Gibbs energy change per molecule (Δg) corresponding to the formation of a spherical cluster of radius r is [30-32]
Δg=4/3πr3vΔμ+4πr2σE2
\n\t\t\t
where v is the molecular volume and σ the interfacial energy per surface. The first term of the right hand equation gives the work produced by the intermolecular attractive forces (Δgb) while the second one corresponds to the work required to increase the surface area in 1cm2 (Δgs). Owing to the meaning of both terms, equation (2) can be rewritten as
Δg=Δgb+ΔgsE3
With the increase of concentration, the amplitude of the fluctuations also increase and so do the size of the clusters. For smaller size clusters, Δgs is the dominant contribution and Δg/dr > 0, i.e., the clusters are unstable. Conversely, for larger size clusters, Δgb is dominant and Δg/dr < 0, meaning that the clusters are stable solid particles. The curve representing the variation of Δg with the radius, shown in Figure 1, passes through a maximum at r = rc (dΔg/dr = 0), with rc representing the limiting size for stable clusters. A cluster of this size is the embryo of the solid form nucleus. The following relationships involving rc can be established:
rc=−2vσΔμarc=−2σΔgbbΔgc=4πrc2σ3cE4
where Δgc corresponds to the value of Δg at rc. The supersaturation concentration (S) can be related with the clusters size by the Gibbs-Thomson equation:
lnCC*=lnS=2vσkTrcE5
\n\t\t\t
Combining (4c) and (5), the following expression is obtained for Δgc as function of the supersaturation:
Δgc=16πσ3v23kTlnS2E6
\n\t\t\t
Δgc represents the energy barrier for the nucleation process and the rate of nucleation (J) will be given by:
J=Ae(−ΔgckT)E7
\n\t\t\t
This equation shows that the rate of nucleation is strongly dependent on the supersaturation and tends to a finite value for a given supersaturation.
Figure 1.
Variation of ∆g and its components with the radius of the molecular clusters.
Organic crystals are supramolecules, i.e, an ensemble of molecules, bonded by non-covalent forces involving ions, polar or non-polar groups. Such interactions, whatever their strength, do not change the molecular individuality. Hydrogen bonding occupies a special place among the intermolecular interactions occurring in molecular crystals. Its energy ranges from 160 kJ mol-1 (strongly covalent) down to 16 kJ mol-1 (mostly electrostatic) [33] and it is highly dependent on the orientation of the hydrogen donor group (D–H) relative to the acceptor atom (A). The maximum strength occurs when the D–H‧‧‧A angle is approximately 180°, though values greater than 110° are acceptable for a hydrogen bond [34].
It is obvious that a crystal packing in which the specific interactions between the functional groups (molecular recognition) are more favorable is, in principle, that leading to the lowest energy. Thus, the lowest energy crystalline structure would correspond to the stronger interaction between a group and its complementar in nature. For example, to a positive charge there would be a negative one, or to a H-bond donor a H-bond acceptor. The molecular packing bringing the molecules close together also gives rise to steric repulsion. Hence, the equilibrium structure is the result between the attraction force and the steric repulsion. This compromise makes the molecule to adopt a conformation corresponding to the lowest Gibbs energy.
Let us consider the conformational variation from the gas phase to the solid for a simple molecule as paracetamol (acetaminophen). This is a drug of great commercial interest in the pharmaceutical industry owing to its wide use as antipyretic and analgesic agent. This molecule exhibits two relevant conformers differing from one another in the orientation of the OH relatively to the carbonyl group [35]. In one of them the dihedral angle formed by the H-O(1)-C-O(2) atoms is close to 180° (trans conformation, Figure 2), while in the other the angle is approximately 0° (cis conformation). Geometry optimization of the two conformers followed by vibrational frequencies calculation at the B3LYP/aug-cc-pVDZ level shows that in the gas phase the population (%) of cis relative to trans is 57:43. Including water effects by applying the CPCM model, both conformers are practically isoenergetic and so their relative population is 50:50. However, in the crystalline phase only the trans conformer is presented. This means that the lattice energy yielded by the trans conformer is more negative than that of the cis conformer. Along this chapter some examples will be given concerning the relationship between the conformers’ population in the three states of matter.
Figure 2.
Optimized geometry of the trans conformer of paracetamol at the B3LYP/aug-cc-pVDZ level.
3. Polymorphism
According to McCrone [36], polymorphism of a compound corresponds to its ability to crystallize into more than one crystalline structure. From the thermodynamic point of view, only the lowest Gibbs energy form exists. However, higher energy polymorphs can remain as metastable forms for a period of time long enough to be used for practical purposes, providing the height of the energy barrier separating these polymorphs and the most stable one is sufficiently high [28]. Since different solid-state modifications exhibit distinct physicochemical properties, such for example the melting point, solubility, dissolution rate and density, polymorphism has a great impact in the pharmaceutical, food and dyes technologies, among others. For example, in the pharmaceutical industry, the desired polymorph for a given active pharmaceutical ingredient (API) would be the one with highest bioavailability and structural stability during shelf life [37].
Figure 3.
Views of the crystalline structures of forms I (a) and II (b) of paracetamol and detail of the H-bonds formed in both polymorphs [(c) and (d)].
A typical example of a drug exhibiting polymorphism is paracetamol. Three polymorphs, labeled as I, II and III, have been identified for this compound [38-40]. The first (form I) is the thermodynamically stable form while the second (form II) is metastable but exists long enough to be experimentally studied. It is usually prepared from cooled melt [41]. Form III is very unstable [41] and thus it is not possible to investigate its structure and properties. Form I crystallizes in the monoclinic system (space group P21/a) [42, 43]. As shown in Figure 3, the molecules in the crystalline structure are linked through O–H O=C and N–H O–H hydrogen bonds forming chains which in turn give rise to pleated layers [frames (a) and (a’)]. Regarding form II, it crystallizes in the orthorhombic system (space group Pcab) [44, 45]. The H-bonding system is identical to that existing in form I but the layers are plane interconnected by van der waals forces [frames (b) and (b’)].
The geometric parameters of the H-bonds in forms I and II are summarized in Table 1. In both forms the O–H‧‧‧O=C hydrogen bond is stronger than the N–H‧‧‧O–H one. In addition, the hydrogen bonds in the stable polymorph are stronger than in the metastable one.
The two polymorphs were also characterized by infrared spectroscopy [46, 47]. The values of the stretching vibration frequencies for the groups involved in hydrogen bonds are given in Table 2. To work as reference, the frequencies of the “free” groups, obtained from a spectrum of paracetamol in a CDCl3 solution were also included in the Table. From the frequency shift (Δν) of the NH and OH groups one can estimate the enthalpy involved in the two hydrogen bonds by applying the empirical relationship proposed by Iogansen: ΔH2 = 1.92(Δν – 40) [48]. The value of ΔH estimated for the N–H‧‧‧O–H H-bond was found to be 12 kJ mol-1 for both polymorphs, while that estimated for the O–H‧‧‧O=C one was found to be 28 kJ mol-1 for form I and 19 kJ mol-1 for form II.
\n\t\t
\n\t\t
\n\t\t
\n\t\t
\n\t\t
\n\t\t
\n\t\t
\n\t\t
\n\t\t\t
Form
\n\t\t\t
\n\t\t\t\tO–H···O=C\n\t\t\t
\n\t\t\t
\n\t\t\t\tN–H···O–H\n\t\t\t
\n\t\t
\n\t\t
\n\t\t\t
H···O/Å
\n\t\t\t
O···O/Å
\n\t\t\t
O–H···O/º
\n\t\t\t
H···O/Å
\n\t\t\t
N···O/Å
\n\t\t\t
N–H···O/º
\n\t\t
\n\t\t
\n\t\t\t
I
\n\t\t\t
1.719
\n\t\t\t
2.653
\n\t\t\t
167.5
\n\t\t\t
2.032
\n\t\t\t
2.904
\n\t\t\t
161.6
\n\t\t
\n\t\t
\n\t\t\t
II
\n\t\t\t
1.838
\n\t\t\t
2.709
\n\t\t\t
170.6
\n\t\t\t
2.069
\n\t\t\t
2.943
\n\t\t\t
163.8
\n\t\t
\n\t
Table 1.
Distances and angles of the hydrogen bonds in the paracetamol polymorphs.a
Vibrational frequencies (ν/cm-1) corresponding to the NH, OH and CO stretching vibrations of paracetamol in the two crystalline phases and in a CDCl3 solution. a\n\t\t\t\t\t
The higher stability of form I is also confirmed by the thermodynamic properties obtained for sublimation, fusion and vaporization of the two polymorphic forms [38]. The value of the packing density, determined by flotation or X-ray (or neutron) diffraction [49] is higher in form II than in form I. In fact, at 298K the density of the former is 1.336 g cm-3 while that of the latter is 1.297 g cm-3. That is, the higher stability of form I results from a stronger hydrogen bond interaction rather than from a more favorable packing. In this compound, the differences between specific interactions resulting from the conformational recognition overcome that due to the crystal packing.
The polymorphism of paracetamol assumes great importance from the practical point of view in so far the commercialized polymorph is not the most suitable solid for formulation [45, 50, 51]. Despite it is easily obtained from various solvents, it has the inconvenient to require a binding agent to make tablets for compression. During this process it gives a brittle solid, consequence of its rough molecular layers. Conversely, form II is constituted by thin plates that glide on pressing. Its plasticity allows the formulation into tablets by direct compression without the need of incorporating any binder agent [45]. The main difficulty to obtain this form comes up against the existence of an adequate method for their preparation at an industrial scale [41].
Aspirin (acetylsalicylic acid) is a drug commonly used as analgesic, antipyretic and anti-inflammatory. It is also known to act as anticoagulant by reducing the blood platelet aggregation [52]. About 35,000 tons of aspirin are taken a day throughout the world.
Using X-ray diffraction, the structure of aspirin has been characterized for the first time in 1964 as a monoclinical crystal, space group P21/c [53]. In 2004, the existence of a second polymorph (form II) was predicted computationally [54] and observed experimentally one year later [55]. However, in view of the slight differences between both structural modifications, it was speculated that this new form might result from uncertainties of the methods used in the structural characterization [56, 57]. Just recently it has been settled the existence of a second polymorph for this compound [58]. In both forms, the carboxyl groups of two neighbor molecules are connected by O–H‧‧‧O=C H-bonds giving rise to centrosymmetric dimers as illustrated in Figure 4. These dimers are arranged into stacked layers, which are identical in the two polymorphs. The acetyl groups of molecules belonging to different layers are in turns interconnected through C–H‧‧‧O interactions. In Form I, these interactions have centrosymmetric geometry, i.e., the CH3 and CO groups of one molecule are bonded to the complementary groups of a neighbor molecule. The structure is a sequence of AAAA layers. Unlike, the structure of form II is a sequence of ABAB layers [59]. The difference between the B and A layers lies in a relative translation of the lattice repetition. This means that in the latter form the CH3 and CO groups of one molecule are connected to the complementary groups of two neighbor molecules, yielding catemers.
Figure 4.
H-bonds in the crystalline structures of forms I and II of aspirin.
An interesting feature of aspirin single crystals is that they exhibit simultaneously domains of forms I and II of variable relative size. Deran et al. [58] explain this type of behavior as an accidental degeneracy. According to these authors the two polymorphs are isoenergetic and during crystallization an intergrowth of both forms occurs. The more favorable internal conformation in form I is compensated by an enhanced cooperativity of the catameric H-bonding network in form II. These results raise the question of whether or not these two forms of aspirin can be called polymorphs. According to the definition given by McCrone [36], they do not fit into the concept.
Besides polymorphism, further crystalline structure complexity arises with the existence of disorder in crystals, situation quite common in many organic crystals [60-64]. This occurs when portions of the structure under analysis occupy two or more positions. For example, molecular fragments of organic molecules not involved in intermolecular interactions can be free enough to acquire different conformations. Such solids are designated by the conflicting denomination of disordered crystals. An example of a compound exhibiting crystal disorder is betaxolol hydrochloride, 1-{4-[2-(cyclopropylmethoxy)ethyl]phenoxy}-3-isopropyl-amino-2-propanol hydrochloride (Figure 5). It is a cardio selective β adrenergic drug that crystallizes into the triclinic system with P1 symmetry [65].
Figure 5.
Conformations of betaxolol hydrochloride in the crystalline structure. Hydrogen bonds between the amino and hydroxyl groups with the chloride ion of the same molecule are represented by dotted lines. For better visualization the hydrogen atoms are not shown.
The intermolecular arrangement in the crystalline structure is as follows: the NH2+ and the OH groups of the isopropylaminoethanol moiety form H-bonds with the chloride ion of the same molecule as shown in Figure 5. In addition, one of the hydrogens of the NH2+ group is also H-bonded to oxygen atom of the OH group of a second molecule while the other hydrogen of the same group is linked to the chloride ion of a third molecule [63, 64]. That is, the betaxolol molecules are connected each other just through the isopropylaminoethanol fragment, leaving a long molecular chain free to move. The X-ray diffraction data shows the existence of an ordered head from the isopropyl group up to O2 and a disordered tail from this atom to the end. By refinement, the crystalline structure can be satisfactory interpreted as being constituted by two conformations which are displayed in Figure 5. Since we have a no well defined unit cell, the structure of betaxolol hydrochloride can not be included in the polymorphism or isomorphism definitions.
4. Erythritol and threitol: identical chemical structure, different crystalline assembling
Erythritol and L-threitol (see Figure 8) are two diastereomers of 1,2,3,4-butanetetrol. The first is a meso compound while the second is optically active, existing as D- or L-threitol. An important difference between both alditols lies in their crystallization ability. Whilst erythritol is easily crystallized from aqueous solution or melt, L-threitol is much more difficult to crystallize [66, 67]. As illustrated in the DSC curves shown in Figure 6, molten erythritol transforms readily into a crystalline phase providing it is cooled at scanning rates lower than or equal to 10°C/min. On the contrary, no crystallization peak is observed for L-threitol, even when the melt is cooled very slowly, say 2°C/min. Since the two compounds are chemically similar, their different crystallization tendency is likely to have a conformational origin [68].
Figure 6.
DSC cooling curves of molten erythritol (m.p = 118°C) and L-threitol (m.p= 89°C) obtained at different cooling rates
Conformationally, both diastereomers are highly flexible. In fact, the existence of seven independent dihedral angles, each of them assuming three standard orientations: gauche+ (60°), gauche– (-60°) and trans (180°), originate a total of 2187 possible conformers. Such a large number of structures make their systematic exploration by means of ab initio calculations unfeasible. Therefore, a previous conformational search method using random generation and subsequent molecular mechanics energy minimization has been employed to sampling the most relevant conformations. All generated structures falling within an energy window of 20 kJ mol-1 above the global minimum have been then optimized at DFT level using the B3LYP functional and the 6-311++G(d,p) basis set. In order to simulate solvent effects, their energy was also evaluated by single-point calculations in aqueous solution using the CPCM continuum model. Details of the computational calculations are given elsewhere [69, 70].
The Boltzmann populations of the erythritol and L-threitol conformers in gas phase and aqueous solution are displayed in Figure 7. The geometries of the relevant conformers in both media, as well as the conformations exiting in the crystalline structures, are depicted in Figure 8. For the sake of simplicity the conformers were grouped according to their backbone family, being labeled with three italic letters: g (gauche+), g’ (gauche–) and t (trans), to specify the orientation of the O(1)-C(1)-C(2)-O(2), C(1)-C(2)-C(3)-C(4) and O(3)-C(3)-C(4)-O(4) dihedrals, respectively.
As shown in Figure 7, isolated erythritol exists preferentially in the tgg and ggg bent conformations (both make up 38% of the conformational mixture at 298.15K). Their stability results from the formation of intramolecular hydrogen bonds which are particularly strong in the tgg conformers due to the participation of the terminal OH groups. In the presence of the solvent, the population of these conformers decreases significantly (11%). Conversely, two higher energy conformers in gas phase (g’tg and gtg), both with a distended carbon backbone, emerge as the most stable forms in solution (49%). This population change can be explained by the stronger hydration of conformers with the OH groups less internally H-bonded and therefore more available to interact with water. In the crystal, the neutron diffraction data available for this compound indicates that the molecule exhibits two conformations, labeled as A and B, differing each other by the positions of the H[O(2)] and H[O(3)] hydrogen atoms. At 22.6 K the occupancy of conformations A and B are 85% and 15%, respectively [71]. In terms of backbone, both have a g’tg conformation which is the same as that exhibited by the preferred conformers of erythritol in aqueous solution. As shown in Figure 8, the geometry of the most stable conformer in solution matches that of the crystal conformation having the highest occupancy percentage.
Figure 7.
Conformational distribution of erythritol and L-threitol in gas phase and aqueous solution. Boltzmann populations were estimated from the Gibbs energies of the conformers [B3LYP/6-311++G(d,p)] in both media.
Regarding L-threitol, its gas phase conformational mixture is largely dominated by the gtg conformers which are characterized by a cyclic, symmetric and cooperative hydrogen bonding network. Since this OH groups’ arrangement is very unfavorable to interact with water, they practically disappear in solution and are replaced by the gg’g’, g’tg’ or g’tg conformations which are not so highly intramolecularly bonded. Unlike erythritol, none of these conformations has been identified in the crystalline L-threitol [72]. Here, the molecules adopt a distended conformation (ttt) which has a relatively low weight in solution (8%) and does not exist in the vacuum.
The results just presented provide an important basis to understand the different crystallization ability of erythritol and L-threitol. In the meso form, this process is facilitated due to the resemblance between the backbone conformation of the molecules existing in the crystal lattice and that characteristic of the predominant forms in solution. The fact that such resemblance is not found to exist in L-threitol may contribute to its lower crystallization tendency. This behavior is in agreement with the results obtained for other alditols [68, 73]
Figure 8.
Most stable conformers of erythritol and L-threitol in gas phase and aqueous solution, as well as the respective crystal conformations. The crystal conformation of erythritol corresponds to that having highest occupancy in the crystal lattice (Conformation A).
5. Conformational variation during the molecular incorporation into the crystal: Glutamic acid
Glutamic acid, 2-aminopentanedioic acid (C5H9NO4), is another example illustrating the role played by conformation on crystallization. Two conformational polymorphs have been identified for this compound, labeled as α and β, with the latter being the thermodynamically stable form over all temperature range [74-77]. The crystals have different morphologies: the metastable form exhibits a prismatic shape while the stable one has a needle-like shape. Both belong to the ortorrombic space group with four molecules in the unit cell (Z=4) [78, 79]. In the crystal lattice the molecules are in the zwitterionic state and all functional groups participate in a complex intermolecular hydrogen bonding network [63].
The conformations adopted by the L-glutamic acid molecules in the two polymorphs are displayed in Figure 9. Their main difference lies in the orientation of the two C-C-C-C dihedral angles. In the crystal lattice the C(1)-C(2)-C(3)-C(4) and C(2)-C(3)-C(4)-C(5) dihedrals assume, respectively, values of 59.2°, 68.3° in α and -171.1°, -73.1° in β. Using the same dihedral labeling scheme previously adopted for the polyols, the conformation of glutamic acid in the α and β-crystals is gg and tg’, respectively. Depending on the experimental conditions, namely the temperature, both polymorphs can be crystallized from aqueous solution. Cooling a supersaturated solution to temperatures below 25°C originates almost pure α-crystals, whereas as the temperature raises the proportion of form β in the precipitated crystals increases. For example at 45°C, the fraction of β-crystals is 45% [74, 75].
The conformational behavior of zwitterionic glutamic acid is aqueous solution is a useful starting point to understand the crystallization of this compound in molecular terms. This has been done theoretically by performing full geometry optimizations using the CPCM continuum solvation model and the B3LYP/aug-cc-pVDZ model chemistry, both implemented in the Gaussian 03 program. The cavity was built with the Bondii radii which have been found to yield accurate results for the hydration of similar molecules in the zwitterionic state, such for example glycine [80]. Nine starting geometries were built by assuming the three standard orientations (g, g’ and t) for each one of the C-C-C-C dihedrals. The remaining dihedrals were kept in their preferred orientations. The optimized structures were further submitted to a vibrational frequency calculation at the same level to ensure that they correspond to minima on the aqueous potential energy surface and also to calculate the thermal corrections at 298.15K.
Figure 9.
CPCM/B3LYP/aug-cc-pVDZ optimized geometries of relevant conformers of the zwitterionic glutamic acid in aqueous solution. Dashed line represents an intramolecular hydrogen bond. Atom numbering scheme is shown for the most stable conformer.
The Gibbs energies of the conformers in aqueous solution at 298.15 K (Gsol) can be expressed as: Gsol = Eint + ΔGthermal + ΔGhyd, with Eint representing the intrinsic (abbreviated as “int”) electronic energy of the conformer at 0 K, i.e., excluding solvent effects, ΔGthermal the thermal correction to the Gibbs energy from 0 to 298K and ΔGhyd the Gibbs energy of hydration at 298.15K. The values of Eint, Gsol, ΔGhyd, as well as the equilibrium populations at 298.15 K, are given in Table 3. In this table are also included the values of the C(1)-C(2)-C(3)-C(3) and C(2)-C(3)-C(4)-C(5) dihedrals, abbreviated as D(1) and D(2), respectively. The relative stability of the conformers in solution is governed by a balance between two effects: interaction with the solvent (quantified by ΔGhyd) and intramolecular interaction (quantified by Eint). In general, conformers with a favorable intramolecular interaction are weakly hydrated and vice-versa. The most abundant conformer in solution, tg (Pop. = 67%), is the one having a less negative ∆Ghyd; however, the intrinsic energy decrease arising from the formation of an internal N+–H‧‧‧O hydrogen bond (see Figure 9), turns it into the lowest energy conformer. Unlike, the second most stable conformer (tt) has a fully distended backbone that hampers the formation of the just referred intramolecular hydrogen bond and so it has a relatively high intrinsic energy. Its stabilization results essentially from a favorable interaction with the solvent. Both conformers represent ca. 81% of the conformational composition in aqueous solution at 298.15K.
Regarding the two crystal conformations, tg’ (β) and gg (α), they are stable forms in aqueous solution with the values of the C-C-C-C dihedral angles in solution not differing significantly from those in the crystal. However, somehow surprisingly, their estimated equilibrium populations in solution are too low: 2.3 and 0.2% for conformers β and α, respectively. Glutamic acid thus constitutes an interesting example of a molecule where the conformers experimentally observed in the crystalline structure are much diluted in aqueous solution. Two immediate conclusions can be drawn from this result: (1) the conformational composition in aqueous solution does not determine the conformations exhibited by this compound in the final crystalline state; (2) crystallization is accompanied by a conformational change. The second conclusion implies that conformers tg and tt, namely the first, are interconverted into the gg and tg’. These interconversions can be characterized by the energy barriers (∆G‡) separating these conformers. For this purpose, we have used the synchronous transit-guided quasi-Newton method (STQN) [81] in its QST2 variety at the CPCM/B3LYP/aug-cc-pVDZ level. The activation energies corresponding to the interconversion of the most stable conformer into conformers α and β were calculated to be ∆G‡ = 18 and 29 kJ mol-1, respectively. This barrier height difference may help to explain the preferential crystallization of form α at temperatures below 25°C and the increase of the percentage of form β in the crystals obtained at higher temperatures. Apparently, the dependence of the crystallization behavior of glutamic acid on temperature is not due to a change on the relative abundance of the crystallizing conformers in solution with temperature [75], but rather to the value of the energy barriers for the interconversion of the most stable conformer in solution into the α and β conformers.
\n\t\t
\n\t\t
\n\t\t
\n\t\t
\n\t\t
\n\t\t
\n\t\t
\n\t\t
\n\t\t\t
\n\t\t\t\tConf.\n\t\t\t
\n\t\t\t
\n\t\t\t\tDihedral angles/º\n\t\t\t
\n\t\t\t
\n\t\t\t\tE\n\t\t\t\tint/ kJ mol-1\n\t\t\t
\n\t\t\t
-ΔG\n\t\t\t\thyd/ kJ mol-1\n\t\t\t
\n\t\t\t
\n\t\t\t\tG\n\t\t\t\tsol/ kJ mol-1\n\t\t\t
\n\t\t\t
Pop.(%)
\n\t\t
\n\t\t
\n\t\t\t
\n\t\t\t\tD(1)\n\t\t\t
\n\t\t\t
\n\t\t\t\tD(2)\n\t\t\t
\n\t\t
\n\t\t
\n\t\t\t
\n\t\t\t\ttg\n\t\t\t
\n\t\t\t
178.5
\n\t\t\t
75.3
\n\t\t\t
13.1
\n\t\t\t
184.8
\n\t\t\t
0.0
\n\t\t\t
67.1
\n\t\t
\n\t\t
\n\t\t\t
\n\t\t\t\ttt\n\t\t\t
\n\t\t\t
168.9
\n\t\t\t
-177.2
\n\t\t\t
56.9
\n\t\t\t
223.7
\n\t\t\t
3.9
\n\t\t\t
13.6
\n\t\t
\n\t\t
\n\t\t\t
\n\t\t\t\tg’g’\n\t\t\t
\n\t\t\t
-52.5
\n\t\t\t
-79.9
\n\t\t\t
0.0
\n\t\t\t
168.2
\n\t\t\t
4.8
\n\t\t\t
9.8
\n\t\t
\n\t\t
\n\t\t\t
\n\t\t\t\tgt\n\t\t\t
\n\t\t\t
63.1
\n\t\t\t
178.8
\n\t\t\t
36.4
\n\t\t\t
196.1
\n\t\t\t
7.4
\n\t\t\t
3.5
\n\t\t
\n\t\t
\n\t\t\t
\n\t\t\t\tg´t\n\t\t\t
\n\t\t\t
-56.5
\n\t\t\t
178.7
\n\t\t\t
52.1
\n\t\t\t
213.7
\n\t\t\t
7.7
\n\t\t\t
3.0
\n\t\t
\n\t\t
\n\t\t\t
\n\t\t\t\ttg’(β)
\n\t\t\t
172.5
\n\t\t\t
-69.8
\n\t\t\t
59.6
\n\t\t\t
221.9
\n\t\t\t
8.4
\n\t\t\t
2.3
\n\t\t
\n\t\t
\n\t\t\t
\n\t\t\t\tgg’\n\t\t\t
\n\t\t\t
62.4
\n\t\t\t
-85.1
\n\t\t\t
49.8
\n\t\t\t
208.1
\n\t\t\t
12.5
\n\t\t\t
0.4
\n\t\t
\n\t\t
\n\t\t\t
\n\t\t\t\tgg(α)
\n\t\t\t
66.1
\n\t\t\t
61.1
\n\t\t\t
54.7
\n\t\t\t
210.5
\n\t\t\t
14.0
\n\t\t\t
0.2
\n\t\t
\n\t\t
\n\t\t\t
\n\t\t\t\tg´g\n\t\t\t
\n\t\t\t
-61.7
\n\t\t\t
81.4
\n\t\t\t
71.1
\n\t\t\t
225.3
\n\t\t\t
18.3
\n\t\t\t
0.0
\n\t\t
\n\t
Table 3.
Dihedral angles, intrinsic electronic energies at 0K (Eint), Gibbs energies in aqueous solution (Gsol), Gibbs energies of hydration (ΔGhyd) and Boltzmann populations (Pop.) at 298.15K of the zwitterionic glutamic acid conformers, calculated at the CPCM/B3LYP/aug-cc-pVDZ level.
6. Molecular dynamics study of pre-nucleation clusters
The molecular aggregates individualized in the early stages of nucleation (pre-nucleation) are windows to follow the crystallization process. Data on the self-assembly of erythritol and L-threitol in supersaturated aqueous solutions are reported through molecular dynamics (MD) simulations. MD is one of the most comprehensive computer simulation tools, with enormous application in the study of large systems and molecular phenomena requiring longer observation times. Relevant molecular phenomena are often adequately described by classical mechanics, providing reliable force-fields are available for a variety of systems. The results of the MD simulations presented in this chapter were carried out in the NpT ensemble and under periodic boundary conditions, resorting to the GROMACS package, version 4.5.4 [82] and GROMOS 96 43a1 force field [83]. Long-range electrostatics was computed using the particle mesh Ewald (PME) method, while for Lennard-Jones energies a cut-off of 1.4 nm was applied. Temperature (298K) and pressure (1bar) were coupled to Berendsen external baths, with coupling constants of 0.1 and 0.5 ps, respectively. Each system was firstly subjected to an energy minimization step, and then left to evolve up to 80ns, using in both parts a standard time step of 2 fs. The last 40 ns of production runs were subsequently subjected to standard analysis, such as radial distribution functions [RDF, g(r)].
Useful data about the type of aggregates formed in solution can be given by the RDFs for the different pairs of solute OH groups (OH–OH) which are depicted in Figure 10. They were obtained from the simulation of an aqueous solution containing 11 solute molecules and 4000 solvent molecules, corresponding to a concentration of 75g /100 cm3, which is near to the saturation concentration [84].
Figure 10.
OH–OH RDFs for erythritol and L-threitol in aqueous solution. For better visualization the most relevant curves are numbered as c1, c2, etc.
In erythritol, the most prominent RDFs are found for the O(2)H–O(3)H and O(1)H–O(4)H pairs. Both RDFs, labeled in Figure 10 as c1 and c2, respectively, exhibit sharp and intense peaks at r (nearest distance between groups) = 0.16 and 0.25 nm. This indicates that the self-assembly of erythritol in solution yields well organized clusters formed by hydrogen bonds involving preferentially the O(1)H and O(4)H groups or the O(2)H and O(3)H groups. Analysis of the MD trajectory shows that these clusters are mostly dimers with some of them involving simultaneously the two just referred H-bonds (cyclic dimers). Interestingly, the cyclic dimer formed by these two H-bonds is the one existing in the crystalline structure. Some illustrative snapshots of these dimers are displayed in Figure 11. From the other OH–OH RDFs shown in Figure 10 one can conclude that the formation of intermolecular hydrogen bonds involving the remaining groups is very unlikely to occur. In fact, the integration of all RDFs up to 0.28 nm (cut-off distance for the formation of an H-bond [85]) reveals that their probability of formation is lower than 15%.
Regarding L-threitol, the RDF profiles are substantially different from those obtained for erythritol. In fact, the most relevant RDFs, labeled in Figure 10 as c1 to c5 and corresponding to the O(3)H–O(3)H, O(2)H–O(3)H, O(2)H–O(4)H, O(1)H–O(3)H and O(2)H–O(2)H interactions, show relatively well defined first peaks centered at 0.15 nm. The first peaks heights are much lower than in erythritol which means that the organized part of the L-threitol clusters is much smaller as compared to that in erythitol. The remaining part of the clusters has a disordered structure as evidenced by the broad peaks located at larger distances. This result is an argument in favor of the lower crystallization tendency of this compound, in addition to the conformational one discussed in section 4. The snapshots of L-threitol clusters during the simulation show the existence of various types of disordered oligomers (Figure 12).
Figure 11.
Snapshots of the erythritol clusters formed in aqueous solution.
Figure 12.
Snapshots of the L-threitol clusters formed in aqueous solution.
7. Conclusion
Structural aspects of crystallization from solvents were pointed out throughout this chapter. Particular attention was paid to the conformational variation of flexible molecules during this process. Three compounds were taken as examples: erythritol, L-threitol and glutamic acid. The different crystallization behavior shown by the two alditols was understood in terms of the resemblance between the solution and crystal conformers. Regarding glutamic acid, it is a quite peculiar example since the conformers existing in the two identified conformational polymorphs have a negligible weight in aqueous solution. Their selective crystallization has been interpreted on the grounds of the energy barriers separating the dominant conformer in solution and those found in both crystalline forms.
The investigation of the structure of the molecular aggregates formed in solution by molecular dynamics, here exemplified for erythritol and L-threitol, has proven to be a valuable contribution to better understand crystallization.
Polymorphism, an important property of the solid state structure with various practical implications, was called in the present chapter as a tangly pathway in the crystallization process. Additional crystalline structure complexity may result from crystal disorder, hardly to be included in the polymorphism concept.
\n',keywords:null,chapterPDFUrl:"https://cdn.intechopen.com/pdfs/43134.pdf",chapterXML:"https://mts.intechopen.com/source/xml/43134.xml",downloadPdfUrl:"/chapter/pdf-download/43134",previewPdfUrl:"/chapter/pdf-preview/43134",totalDownloads:3221,totalViews:519,totalCrossrefCites:3,totalDimensionsCites:3,totalAltmetricsMentions:0,impactScore:1,impactScorePercentile:62,impactScoreQuartile:3,hasAltmetrics:0,dateSubmitted:"May 16th 2012",dateReviewed:"October 16th 2012",datePrePublished:null,datePublished:"February 20th 2013",dateFinished:"February 16th 2013",readingETA:"0",abstract:null,reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/43134",risUrl:"/chapter/ris/43134",book:{id:"3349",slug:"advanced-topics-on-crystal-growth"},signatures:"J.S. Redinha, A.J. Lopes Jesus, A.A.C.C. Pais and J. A. S. Almeida",authors:[{id:"80738",title:"Dr",name:"António Jorge",middleName:null,surname:"Lopes Jesus",fullName:"António Jorge Lopes Jesus",slug:"antonio-jorge-lopes-jesus",email:"ajorge@qui.uc.pt",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:null}],sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_2",title:"2. Nucleation and crystal growth",level:"1"},{id:"sec_3",title:"3. Polymorphism",level:"1"},{id:"sec_4",title:"4. Erythritol and threitol: identical chemical structure, different crystalline assembling ",level:"1"},{id:"sec_5",title:"5. Conformational variation during the molecular incorporation into the crystal: Glutamic acid ",level:"1"},{id:"sec_6",title:"6. Molecular dynamics study of pre-nucleation clusters ",level:"1"},{id:"sec_7",title:"7. Conclusion",level:"1"}],chapterReferences:[{id:"B1",body:'Brian Henderson and Ralph HB. Crystal-Field Engineering of Solid-State Laser Materials. 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The crystal and molecular structure of aspirin. Journal of the Chemical Society (Resumed). 1964:6036-48.'},{id:"B54",body:'Ouvrard C, Price SL. Toward Crystal Structure Prediction for Conformationally Flexible Molecules: The Headaches Illustrated by Aspirin. Crystal Growth & Design. 2004;4(6):1119-27.'},{id:"B55",body:'Vishweshwar P, McMahon JA, Oliveira M, Peterson ML, Zaworotko MJ. The Predictably Elusive Form II of Aspirin. Journal of the American Chemical Society. 2005 2012/09/10;127(48):16802-3.'},{id:"B56",body:'Bond AD, Boese R, Desiraju GR. On the Polymorphism of Aspirin. Angewandte Chemie International Edition. 2007;46(4):615-7.'},{id:"B57",body:'Chang C-J, Díaz LE, Morin F, Grant DM. Solid-state 13C NMR study of drugs: Aspirin. Magnetic Resonance in Chemistry. 1986;24(9):768-71.'},{id:"B58",body:'Wen S, Beran GJO. Accidental Degeneracy in Crystalline Aspirin: New Insights from High-Level ab Initio Calculations. Crystal Growth & Design. 2012 2012/09/10;12(5):2169-72.'},{id:"B59",body:'Bond AD, Boese R, Desiraju GR. What Is A Polymorph? Aspirin As A Case Study. American Pharmaceutical Review. 2007:1-4.'},{id:"B60",body:'Glusker JP, Lewis M, Rossi M. Crystal Structure Analysis for Chemists and Biologists (Methods in Stereochemical Analysis). New York: Wiley-VCH; 1994.'},{id:"B61",body:'Wilson CC. A basic introduction to thermal motions of atoms in crystal structures, the underlying potentials and the physical information available from their analysis. Crystallography Reviews. 2009 2012/09/11;15(1):3-56.'},{id:"B62",body:'Habgood M, Grau-Crespo R, Price SL. Substitutional and orientational disorder in organic crystals: a symmetry-adapted ensemble model. Physical Chemistry Chemical Physics. 2011;13(20):9590-600.'},{id:"B63",body:'Redinha JS, Lopes Jesus AJ. Molecular Recognition and Crystal Growth. In: McEvoy JA, editor. Molecular Recognition: Biotechnology, Chemical Engineering and Materials Applications: Novapublishers; 2011.'},{id:"B64",body:'Redinha JS, Lopes Jesus AJ. Crystal Growth of Pharmaceuticals from Melt. In: Borisenko E, editor. Crystallization and material science of modern artificial and natural crystals. Rijeka: InTech; 2012.'},{id:"B65",body:'Pascard C, Tran Huu Dau E, Manoury P, Mompon B. Betaxolol hydrochloride: 1-{4-[2-(cyclopropylmethoxy)ethyl]phenoxy}-3-isopropylamino-2-propanol hydrochloride, C18H30NO3+.Cl. Acta Crystallographica, Section C: Crystal Structure Communications. 1984;40(8):1430-2.'},{id:"B66",body:'Lopes Jesus AJ, Nunes SCC, Ramos Silva M, Matos Beja A, Redinha JS. Erythritol: Crystal growth from the melt. International Journal of Pharmaceutics. 2010;388(1-2):129-35.'},{id:"B67",body:'Lopes Jesus AJ, Redinha JS. On the structure of erythritol and L-threitol in the solid state: An infrared spectroscopic study. Journal of Molecular Structure. 2009;938(1-3):156-64.'},{id:"B68",body:'Yu L, Reutzel-Edens SM, Mitchell CA. Crystallization and Polymorphism of Conformationally Flexible Molecules: Problems, Patterns, and Strategies. Organic Process Research & Development. 2000 2012/09/14;4(5):396-402.'},{id:"B69",body:'Lopes Jesus AJ, Tomé LIN, Rosado MTS, Leitão MLP, Redinha JS. Conformational study of erythritol and threitol in the gas state by density functional theory calculations. Carbohydrate Research. 2005;340(2):283-91.'},{id:"B70",body:'Lopes Jesus AL, Tomé LIN, Eusébio MES, Redinha JS. Determination of the Enthalpy of Solute-Solvent Interaction from the Enthalpy of Solution: Aqueous Solutions of Erythritol and L-Threitol. Journal of Physical Chemistry B. 2006 May 11, 2006;110(18):9280-5.'},{id:"B71",body:'Ceccarelli C, Jeffrey GA, McMullan RK. A Neutron-Diffraction Refinement of the Crystal-Structure of Erythritol at 22.6 K. Acta Crystallographica Section B. 1980;36:3079-83.'},{id:"B72",body:'Kopf J, Morf M, Zimmer B, Haupt ETK, Jarchow O, Köll P. The crystal and molecular structure of threitol. Carbohydrate Research. 1993;247:119-28.'},{id:"B73",body:'Jeffrey GA, Kim HS. Conformations of the alditols. Carbohydrate Research. 1970;14(2):207-16.'},{id:"B74",body:'Kitamura M. Polymorphism in the crystallization of L-glutamic acid. Journal of Crystal Growth. 1989;96(3):541-6.'},{id:"B75",body:'Kitamura M. Controlling factor of polymorphism in crystallization process. Journal of Crystal Growth. 2002;237-239:2205-14.'},{id:"B76",body:'Davey RJ, Blagden N, Potts GD, Docherty R. Polymorphism in Molecular Crystals: Stabilization of a Metastable Form by Conformational Mimicry. Journal of the American Chemical Society. 1997;119(7):1767-72.'},{id:"B77",body:'Ono T, ter Horst JH, Jansens PJ. Quantitative Measurement of the Polymorphic Transformation of l-Glutamic Acid Using In-Situ Raman Spectroscopy. Crystal Growth & Design. 2004;4(3):465-9.'},{id:"B78",body:'Lehmann M, Koetzle T, Hamilton W. Precision neutron diffraction structure determination of protein and nucleic acid components. VII. The crystal and molecular structure of the amino acid L-lysine monohydrochloride dihydrate. Journal of Chemical Crystallography. 1972;2(5):225-33.'},{id:"B79",body:'Lehmann MS, Nunes AC. A short hydrogen bond between near identical carboxyl groups in the [alpha]-modification of l-glutamic acid. Acta Crystallographica, Section B: Structural Science. 1980;36(7):1621-5.'},{id:"B80",body:'Kim CK, Park B-H, Lee HW, Kim CK. Comprehensive Studies on the Free Energies of Solvation and Conformers of Glycine: A Theoretical Study. Bulletin of the Korean Chemical Society. 2011;32(6):1985 - 92.'},{id:"B81",body:'Peng CY, Ayala PY, Schlegel HB, Frisch MJ. Using redundant internal coordinates to optimize equilibrium geometries and transition states. Journal of Computational Chemistry. 1996;17(1):49-56.'},{id:"B82",body:'Hess B, Kutzner C, van der Spoel D, Lindahl E. GROMACS 4: Algorithms for Highly Efficient, Load-Balanced, and Scalable Molecular Simulation. Journal of Chemical Theory and Computation. 2008;4(3):435-47.'},{id:"B83",body:'Schuttelkopf AW, van Aalten DMF. PRODRG: a tool for high-throughput crystallography of protein-ligand complexes. Acta Crystallographica Section D. 2004;60(8):1355-63.'},{id:"B84",body:'Cohen S, Marcus Y, Migron Y, Dikstein S, Shafran A. Water sorption, binding and solubility of polyols. Journal of the Chemical Society, Faraday Transactions. 1993;89(17):3271-5.'},{id:"B85",body:'Gilli G, Gilli P. Towards an unified hydrogen-bond theory. Journal of Molecular Structure. 2000;552(1-3):1-15.'}],footnotes:[],contributors:[{corresp:null,contributorFullName:"J.S. Redinha",address:null,affiliation:'
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1. Introduction
Global warming is one of the major challenges for mankind, with animal breeding one of the most affected sectors in the agricultural industry. The impacts of increasing environmental temperatures on livestock will most likely differ from place to place, depending on latitude, geographical features and local farming systems [1, 2, 3].
High ambient temperatures negatively affect all domestic animals, but in addition to pork and dairy production, perhaps the poultry industry is hit the hardest. In 2020, the world’s broiler meat production amounted to about 100.81 million metric tons, and is forecasted to increase to about 101.02 million metric tons by 2021 [4]. According to FAO data [5], total egg production in the world was 1.528 billion units in 2018. In 2019, this figure reached 1.577 billion.
These statistics clearly show that broiler meat and egg production play a crucial role in the global supply of animal origin foodstuffs.
Thus, we have a fundamental interest in reducing and/or eliminating the negative effects of climate change, i.e. prolonged high ambient temperature. The main question is, what tools do we have to reduce the harmful effects of high environmental temperatures-especially in the case of heat stress? A solution for prevention of heat stress in animals includes biological (e.g. genetics, thermal conditioning, nutrition) [6, 7] or keeping technology devices (e.g. air conditioning, intensive ventilation, humidification) [8]. However, housing methods are expensive and the service costs are high. Therefore, reducing the biochemical and physiological negative effects of heat stress with different nutritional tools is one of the primary interests for the economical production of food produced from animals.
According to Babinszky et al. [9], basically the following nutritional possibilities are available to eliminate the harmful effects of the heat stress:
reduce animal’s own heat production (e.g. feeding more dietary fat);
compensate for the lower nutrient supply; (e.g. feeding more concentrated diets); and
mitigate heat stress induced metabolic changes (e.g. using different feed additives: vitamins, micro minerals).
It should, however, be noted that during severe heat stress, these methods should be used in combination in order to maintain the production performance of the farm animals and the quality of their products [9]. While this chapter focuses on the third option, i.e. the use of feed additives, we would like to emphasize that whatever feeding method we use, we need to be aware of the changes in the intermediate metabolism of farm animals caused by heat stress, because without this knowledge, there is no effective defense against high ambient temperatures.
Therefore, the aim of this chapter is to summarize the adverse effects of heat stress on energy metabolism, anti- and pro-oxidant capacity, and production in birds. A further goal is to show how various feed additives (vitamin A, C and E, selenium, zinc, betaine, plant extract, and probiotics) can reduce the negative effects of heat stress.
2. Methodology of the literature review
The methodology of the literature review was basically the same as the internationally applied methodology used in animal science. Firstly relevant literature was searched. This follows by evaluation of sources. The third step was identifying the database and gaps in the published scientific findings, than setup the outline structure. Finally the literature review was written.
The literature searching was based on the keywords, using university database, own department data collection on the research field of heat stress, and different international scientific databases of life sciences, animal science and Google Scholar.
In each of the studied paper or book chapter, we asked the same questions as, for example:
What was the aim and methodology of the particular publication (in this case: what kind of heat stress was applied, how many animals were included in the experiment per treatment, whether there were repetitions, what dietary treatments (type of feed additives and their concentration in the diet) were used, what parameters were measured, what was the statistical analysis applied, etc.), furthermore, whether experimental data were correctly evaluated, what results were presented by the authors and what main conclusions were drawn from the data.
To have more clear information on effectiveness of various feed supplements in case of production parameters: daily gain (g/d), average daily gain (g/d) and feed conversion ratio (kg diet/kg gain), the so called mitigation capacity was calculated using the following formula:
Mitigation capacity ofacertain trait%=measured value inHSenvironment andfedwiththe experimental diet−measured value inHSenvironment andfedwith the control diet/measured value inTNenvironment andfedwith the control diet–measured value inHSenvironment andfedwith the control dietx100E1
where: HS = heat stress; TN = thermoneutral.
All collected information (data) was placed in a large work database. This information formed the basis of the subchapter titles of our review chapter and of the chapter outline. Based on this information, the evaluation of research data from more than 90 publications started. The writing of the review chapter then began, including the drawing of main conclusions as well. The investigated and systematized research findings are summarized in tables.
3. Heat production of animals and heat stress
It is well known that heat production of animals is the sum total of non-productive energy utilized by the animal and of the energy lost in the course of transformation dietary nutrients [10]. Animals use this so called non-productive energy for maintenance (i.e. satisfy the energy requirement for the maintenance of body temperature, the functioning of the nervous system, the organs, for minimal activity, etc.) [10]. The extra heat produced in the course of digestion, excretion and metabolism of nutrients is called the heat increment. It is also well known that within a certain range of ambient temperature - with unvarying feed and nutrient intake - the total heat production of the animal remains constant. This temperature range is called the thermoneutral zone. The general scheme of the relationship between ambient temperature and heat production of livestock can be seen in Figure 1 [10].
Figure 1.
Relationship between ambient temperature and heat production of livestock [10].
In a thermoneutral environment, the heat production of the animal is at the minimum, and thus the dietary energy can be used for production (growth, egg and milk production) efficiently [9, 10]. Therefore, whenever the daily amount of energy intake changes, the temperature range of the thermoneutral zone is changed, too. So, if for some reason the animal leaves the thermoneutral zone, this result in an increased heat production by the animal. This means that there is more loss of energy, and in consequence, less energy remains for production and moreover the efficiency of energy utilization deteriorates too. The upper and lower critical temperatures for poultry are summarized in Table 1 [11].
The general scheme of the relationship between broiler behavior and the increasing ambient temperature is shown in Figure 2 [12].
Figure 2.
The effect of increasing ambient temperature on birds [12].
As can be seen in Figure 2, in the thermoneutral zone, birds can lose heat at a controlled rate using normal behavior [12]. Between the lower and upper temperatures, there is no heat stress and body temperature remains constant. If the environmental temperature exceeds the upper critical temperature, birds must lose heat actively by panting. However, it should be noted that panting is a normal response to heat and is not initially considered a welfare problem [12]. However, as temperatures increase, the rate of panting increases. If heat production is greater than maximum heat loss, birds may die due to heat stress. In other words, heat stress occurs when the body cannot get rid of excess heat.
It is well established that heat stress increases the energy cost of maintenance and adversely affects productive and reproductive performance. In a hot environment, the respiration rate in birds can increase 10–20 times, causing increased CO2 loss through the lungs [13]. This loss results in an increase in blood pH and this can upset the acid–base balance, which can impair the health and performance of birds [14, 15, 16].
There are usually two types of heat stress, acute and chronic heat stress. Acute heat stress refers to a short and rapid increase in environmental temperature (a few hours), whereas under chronic heat stress, high temperatures persist for more extended periods (several days) [17].
Heat stress exposed animals can use different ways to maintain thermoregulation and homeostasis. They can increase radiant, convective and evaporative heat loss by vasodilatation and perspiration [18]. However, birds have an extra mechanism which is promote heat exchange between their bodies and the environment. These are the air sacs. Air sacs are very useful especially during panting, as they promote air circulation on surfaces and consequently, the evaporative loss of heat [19, 20].
Unfortunately, there are only few scientific papers that report on the heat production and the heat loss of heat-exposed birds. Consequently, there is only a limited number of scientific publications that report on nutritional possibilities for reducing the heat production of birds under heat stress.
Syafwan et al. [21] concluded in their excellent review that the heat production of broilers is particularly high due to the high growth rate and the high daily feed intake. Developments in the genetic selection of meat-type birds has led to rapid growth and a high metabolic rate, which is accompanied by a higher heat production level due to increased feed intake [22]. Therefore, it can be stated that high genetic capacity hybrid broilers (so called “improved chicken”) are much more sensitive to a hot environment than their unimproved counterparts.
Summarizing the relevant scientific findings, it can be stated that in practical animal agriculture, and especially in factory farming, it is particularly difficult to keep animals in a thermoneutral zone. Therefore, in order to reduce the negative effect of heat stress, it is important to use nutritional tools in addition to technical devices.
4. Reduction of heat production by nutritional tools
4.1 Using fat in the diets
It is well known that if more fat is used in pig diets in high ambient temperature, the total heat production of the animals reduces significantly. Babinszky et al. [23] concluded from their study that lactating sows fed a high level of dietary fat (125 g fat/kg diet) produce significantly less heat than those fed a carbohydrate rich (low-fat level) diet. Babinszky [10] is also concluded, that the energetic efficiency of milk production was improved, when sows received high dietary fat diet (125 g/kg diet). This phenomenon can be explained by the fact that synthesizing milk fat from dietary fat is more efficient than it’s synthesizing from dietary carbohydrates.
In poultry nutrition, relative limited literature data are available on fat feeding against heat stress and its effect on heat production of birds. Das et al. [24], in an excellent review, stated that heat stress may be combated by adding fat and reducing crude protein in poultry diets. Higher energy diets were effective in partially mitigating the effects of heat stress in poultry. This can be explained by the fact that during metabolism, fat produces a lower heat increment than protein and carbohydrates [25].
In other studies, it was concluded that supplementation of fat in the poultry diet increase the nutrient utilization in the gastrointestinal tract by lowering the rate of food passage [26] and also helps increase the energy value of the other feed constituents [27, 28]. Feeding a high fat diet (up to 5%) to heat-exposed broilers reduces heat production. This result occurs because the heat increment of fat is lower than that of either proteins or carbohydrates [21, 25, 29, 30].
4.2 Using vitamin C in chicken diet to change energy metabolism
Because the animal body derives all its energy from oxidation, the magnitude of energy metabolism can be determined from the amount of carbon-dioxide produced and oxygen consumed. The ratio of the volumes of carbon dioxide produced to oxygen consumed is called the respiratory quotient (RQ) [31]. The respiratory quotients are: for protein: 0.809; for fat: 0.711; for starch: 1.000; for sugar: 1.000; and for glucose: 1.000, respectively [32]. If the RQ value is equal to 1.00, this means that e.g. burning 1 g of starch produces as much carbon dioxide as oxygen is needed to burn it (0.829 liter CO2/0.829 liter O2).
However, it should also be noted that RQ values significantly higher than 1 can be achieved if the animals convert the carbohydrate to fat, since in this case oxygen-poor fat is formed from oxygen-rich glucose. During starvation, the RQ value is less than 0.7 [31].
As it can be seen above, the RQ may provide valuable information about the metabolic processes in the body. Therefore, RQ values are very often determined in respiratory studies.
McKee et al. [33] investigated the effect of vitamin C on different variables of energy metabolism of young heat exposed chickens in indirect calorimeters. The experiment started at day 9 and lasted until day 17 posthatch. In this study, CO2 production and O2 consumption were measured in the thermoneutral zone (27.7°C) and in a hot environment (34°C). On the basis of these values, RQ and heat production were calculated daily through day 17 of the experiment. The basal diet was supplemented by a 150 mg/kg diet of ascorbic acid (vitamin C). They found that heat exposure lowered (P < 0.001) the respiratory quotient. Heat-exposed birds consuming the ascorbic acid supplemented diet expressed lower respiratory quotients than their unsupplemented counterparts. The authors concluded that this effect resulted from a nonsignificant increase in O2 consumption and decrease in CO2 production. They also concluded that further investigations are needed to determine whether the ascorbic acid-induced change in the RQ value towards 0.70 reflects an increase in protein or lipid catabolism or both. In further study, the effect of ascorbic acid on the energy metabolism and heat production of domestic animals should also be elucidated.
Despite the many open questions, based on the findings made by McKee et al. in their study, it seems that supplemental ascorbic acid may influence the body energy stores during periods of reduced energy intake (during heat stress).
4.3 Using betaine as feed additive in the diet
Betaine chemical structure (C5H11NO2) contains three methyl groups which play a role in transmethylation reactions [34]. Betaine (trimethylglycine) is an intermediate metabolite in the catabolism of choline which can modify osmolarity, act as a methyl donor, and has potential lipotropic effects [9]. As a by-product of sugar beet processing, betaine is commercially available as a feed additive [35]. Currently, betaine is available in several purified forms (anhydrous, monophosphate and hydrochloride betaine) [36].
Betaine mainly functions as an osmolyte and a methyl-group donor [37]. Under heat stress, betaine plays an important role in cellular osmotic regulation, preventing dehydration by increasing the water-holding capacity of cells. It helps in maintaining the protective osmolytic activity in birds under heat stress. Betaine may promote various intestinal microbes against osmotic variations and this results in improve microbial fermentation activity [38]. Furthermore, betaine is also found to have anti-inflammatory properties and improves intestinal function [39].
Because betaine influences fat and protein deposition, it can also be used to improve carcass quality and reduce fatty livers. Schrama et al. [40] showed that energy retention in pigs improves over time following the supplementation of betaine to the diet. They also found that under thermoneutral conditions, dietary betaine supplementation (1.23g/kg diet) reduced the total heat production of pigs. They suggest the same concentration of betaine in poultry diet, as well.
These scientific findings suggest that betaine may be suitable for reducing heat production in livestock (e.g. in poultry) at high ambient temperatures. However, only few research results have been published in this area to date. Therefore, further studies are needed to determine the impact of betaine on heat production in animals under high ambient temperature.
Another problem is that many of published papers are not clear on the source of betaine used (natural, extracted betaine anhydrous or synthetic betaine hydrochloride). The source is important, as it is likely that the efficacies of the different betaine sources differ.
5. Effects of heat stress on anti- and prooxidant status in birds. Mitigation using different feed additives
5.1 Anti- and prooxidant status
5.1.1 Impacts of heat stress
Increased environmental temperature caused increased lipid peroxidation (as well as induced formation of malondialdehyde (MDA), which is an indicator for lipid peroxidation). Therefore, the antioxidant defense system is altered [41, 42, 43].
According to the latest research, the elimination of the free radicals activates three level antioxidant systems (Figure 3, based on Babinszky et al. [10]).
Elimination is done by the first level of the antioxidant system which functions at the same time as the detoxification and regeneration pathways of the second level. The third level starts working after damage has been done, to repair and eliminate damaged cells. This first level (direct enzymatic pathway) includes the neutralization of the oxygen and nitrogen centred free radicals by enzymes. The second level includes the detoxification and regeneration reactions of the small molecule antioxidants. The third level is activated when damaged systems (proteins, DNA) have to be repaired and/or removed from the cells by chaperones and DNA-repair enzymes.
In general, it can be concluded that a large amount of Reactive Oxygen Species (ROS) causes disruption of mitochondrial function, increased lipid peroxidation, and decreased the concentration of so called antioxidant vitamins, furthermore induce stress gene expression, and finally it leads to dysfunction in antioxidant enzymes and also causes DNA damage.
According to Yang et al. [44] in heat stressed broilers (35°C for 3h/day), the activity of the mitochondrial respiratory chain is reduced, which led to over-production of ROS. This situation results in lipid peroxidation and oxidative stress in the birds.
In another study [7] lipid peroxidation and superoxide dismutase (SOD) activity was measured in broilers under heat stress (32°C for 6 h/day). The results showed that high temperature disturbed the equilibrium between the synthesis and catabolism of ROS production. Glutathione peroxidase (GPx) and SOD activity increased and catalyze (CAT) activity decreased under heat stress (34°C 5 h/day from d28 to d38) [45].
ROS production reduced Vitamin A and E levels. Vitamin C concentration decreased under heat stress in poultry [46]. It has been reported that heat stress increased zinc (Zn) mobilization from tissues, and thus may cause marginal Zn deficiency and increase requirements [47]. According to Zeng et al. [48, 49], SOD, MDA, CAT activity and the total antioxidant capacity (T-AOC) in Muscovy duck liver increased under short term heat stress (39°C for 1 hour then 3-hour recovery at20°C). The same results were found in broilers [50]. During heat stress in broilers, the serum concentrations of Vitamin C, E, A, iron (Fe), and Zn decreased, while the copper (Cu) concentration increased [51].
5.1.2 Using feed additives
5.1.2.1 Vitamin supplementation
High environmental temperature decreases the concentrations of vitamins and micro minerals in serum and increases excretion [52]; therefore, supplementation of direct or indirect antioxidant compounds (e.g. vitamins and micro nutrients) at higher levels is commonly recommended. These additives support mechanisms against lipid peroxidation, improve immune status and performance.
5.1.2.1.1 Vitamin E
Vitamin E functions as a fat-soluble antioxidant which protects cellular and membrane lipids from peroxidation-catalyzed free radicals due to heat stress. In cell membranes and lipoproteins, the essential antioxidant function of Vitamin E is to trap ROO− and to break the chain reaction of lipid peroxidation. While it cannot prevent their formation, it can reduce the formation of secondary radicals [53]. Vitamin E is known as the first line of defense against lipid peroxidation caused by heat stress. It has free radical quenching activity and attacks free radicals in an early stage. When feed was supplemented with Vitamin E (200-250 mg/kg feed), the serum concentration of Vitamins E and A increased in serum and MDA concentration decreased under long term heat stress [51]. Maini et al. [54] reported that CAT, GR, GSH, MDA and SOD level decreased under heat stress due to Vitamin E supplementation. Short term heat stress increased the concentration of Zn in serum when the diet was supplemented with Vitamin E [55] (Table 2, [56]).
Effects of selected vitamin supplementation under heat stress based on different studies [56].
CAT = catalase concentration in blood; GR = glutathione reductase concentration in blood; GSH = reduced glutathione concentration in blood; MDA = malondialdehyde concentration in blood; SOD = superoxide dismutase concentration in blood; Zn = zinc.
↑ Increase; ↓ decrease.
5.1.2.1.2 Vitamin C
Vitamin C protects against oxidative stress-induced cellular damage in the presence of scavenging ROS, and is capable itself of inhibiting lipid peroxidation in plasma. Ascorbic acid can directly scavenge radicals in the aqueous compartment. Ascorbate can scavenge O2−, H2O2, the OH, hypochlorous acid, aqueous ROO−, and singlet oxygen. Under its antioxidant activity, ascorbate has a two-electron reduction [53]. Although chickens are known to synthesize ascorbic acid in the kidney, increased supplementation has proved beneficial effects in broilers reared under heat stress [59]. Ascorbic acid is actively absorbed. This active transport is supported by the sodium electrochemical gradient. However, the vitamin C requirements increase under heat stress. According to different studies, ascorbic acid supplementation (200 mg/kg feed) caused a significant increase in plasma ascorbic acid levels [59, 60] in broilers under heat stress. This indicates that the higher Vitamin C concentrations in the broiler diet could be used against heat stress successfully.
5.1.2.2 Micro-mineral supplementation
5.1.2.2.1 Zinc
Zn is a “member” of the antioxidant network because it is a cofactor of a very important antioxidant enzyme: Cu/Zn-SOD. Zinc plays a role in depressing the free radicals and inhibiting lipid peroxidation and GSH depletion. Zn can have direct antioxidant function and it is necessary for the prevention of free radical formation. However, it does not act directly against them [53]. Zinc supplementation has positive effects on antioxidant status of birds [61, 62, 63]. Zinc may play an important role in suppressing free radicals because it works as a cofactor (Cu/Zn-SOD) and inhibits NADPH-dependent lipid peroxidation [64], thus improving antioxidant status: increased serum Vitamin C and E concentrations [65] and decreased MDA levels [57, 66] (Table 3, [56]).
Effects of micro mineral supplementation under heat stress based on different studies [56].
GPx = glutathione peroxidase concentration in blood; GR = glutathione reductase concentration in blood; GSH = reduced glutathione concentration in blood; GSSG = glutathione disulphide concentration in blood; GPx = glutathione peroxidase concentration in blood; MDA = malondialdehyde concentration in blood; Se = selenium; Zn = zinc.
↑ Increase; ↓ decrease; ᴓ no effect.
5.1.2.2.2 Selenium
Organoselenium compounds are essential micronutrients and are required for cellular defense against oxidative stress and optimal immune function. Selenium is necessary for cellular function and is a component of antioxidant enzymes: an important part (cofactor) of GPx, which works as an important antioxidant enzyme, protecting cells against free radical damage and oxidative stress [53]. Selenium supplementation improved antioxidant status in poultry under heat stress [51, 55, 58]. It is suggested that the metabolic role of Se is to protect cells against oxidation and tissue damage. Rapid oxidation of GSH to GSSH is necessary to compensate the heat stress caused ROS production. However, Se supplementation increases the level of available NADPH to promote the activation of GR, leading to increased GSSH reduction to GSH [67]. Therefore, Se supplementation affected GPx activity and the GPx/GSH ratio (Table 3).
Results of studies done with separated supplementation of Vitamin A (9000–15000 IU/kg diet), Vitamin E (150–500 mg/kg diet), Vitamin C (150–500 mg/kg diet), Zn (30 or 60 mg/kg diet) and Se (0,1-1 mg/kg diet), show that antioxidant status improved in poultry under heat stress. Antioxidant potential has been reported to be more efficient and important in combination than single antioxidant nutrients [68]. The latest research studies show that interactions between vitamin-vitamin and vitamin-minerals used in combination have more improved effects on the antioxidant status and performance of poultry under heat stress than they do separately. Literature data on combinations of vitamin and mineral supplementation can be seen in Table 4 [56].
Effects of vitamin and mineral interactions under heat stress based on different studies [56].
MDA = malondialdehyde concentration in blood; SOD = superoxide dismutase concentration in blood, Se = selenium, Zn = zinc.
↑ Increase; ↓ decrease.
5.1.2.3 Plant extracts
Dried oregano powder (0.5% and 1%) can be supplemented for ducks. Oregano (Origanum vulgare L.) is an herb extract used as an additive in poultry nutrition. It is an aromatic plant, containing more than 30 phenolic antioxidants constituents, including also anti-inflammatory and anti-microbial activity. It can also have beneficial effects on production, mortality, microflora, and the immune system [69]. Antioxidant enzyme activity (SOD, GPx) was improved in poultry [69]. These results suggest that dried oregano powder addition could decrease the changes in antioxidant enzymes under heat stress.
5.1.2.4 Probiotics
Probiotics are non-digestive alternative growth promoters used in poultry nutrition. Probiotics can improve animal performance, and it can manipulate and maintain beneficial microflora in the gut. Several studies prove that probiotics supplementation to feed improves production parameters in poultry [70, 71]. Supplementation of probiotics (Bacillus Subtilis: 1x108 CFU/kg feed) decreased MDA activity and uric acid concentration, and also improved antioxidant response in ducks [72, 73].
6. The effect of heat stress on the performance of broilers reduced by using feed additives
6.1 The effect of heat stress on the performance of broilers
After reviewing the relevant research, we identified three different types of heat stress (HS) that have been applied in experiments: acute, cyclic and chronic. In the case of acute HS, the elevated temperature lasts from several hours up to 24 hours. After exposure to HS, sample and data collection occurs. This type of arrangement is suitable to study the immediate effect of heat stress. However, in temperate countries, even in the case of cooled stables, the actual barn temperature shows a daily cycle which can be mimicked by the cyclic HS environment (4–10 hours per day in the range of three days per week to daily up to 10 days long). In tropical countries, this kind of fluctuation is much less pronounced. Therefore, the environmental conditions can be best modeled with a chronic HS (continuous HS environment usually during the second half of fattening) model [74].
The most often claimed effect of heat stress is a reduction of feed intake. As an immediate effect, acute heat stress reduces feed intake by about 25% (Table 5).
Effect of heat stress on the growth performance of broilers (means ± s.d.).
Thermoneutral environment.
Heat stress environment.
(TN-HS)/TN*100, average values of the calculations from the research cited.
HS environment from several hours to 24 hours.
HS environment 4–10 hours per day in the range of three days per week to daily up to 10 days.
Continuous HS environment (24 hours/day) usually during the second half of the fattening.
Applying the heat stress repeatedly but allowing regeneration at thermoneutral (TN) temperature (cyclic HS-simulating the day-night temperature fluctuation) results in an adaptation, as during this period the lowest decline (in between 5 and 15%) in performance data (feed intake, daily gain and feed conversion ratio) can be observed (Table 5). Chronic heat stress will approximately double the negative effect compared to cyclic heat stress (7–11% point), but still some adaptation can be seen compared to acute heat stress. Acute HS has a dramatic effect on daily gain, as even negative values (weight loss) can occur, which makes it impracticable to calculate the feed conversion ratio. Therefore, researchers did not publish such data. The nutrient content of the unconsumed feed itself does not justify the negative energy balance; therefore, one can assume that the energy and nutrient needs of the HS response are high. When adaptation can occur; cyclic HS has a less adverse effect than chronic HS on both daily gain and the feed conversion ratio (Table 5).
6.2 Mitigation capacity of various feed additives on HS in broilers
One long term aim of researchers is to be able to mitigate the negative effects of heat stress. Supplementation of effective feed additives could be useful for improving intestinal absorption and minimizing the adverse effects of HS [92]. To have more clear information on effectiveness of various feed supplements Ortega and Szabó [93] suggested the calculation of mitigation capacity (see subsection 2 in the present chapter). However, researchers [93] also point out that contrary to the numerous publications in the field, only a limited number of studies are suitable for using this calculation, as it requires at least three treatment groups. HS effect mitigating supplements are usually vitamins expressing antioxidant capacity, probiotics and plant extracts (Tables 6–8).
Mitigation capacity of various feed additives on HS in broilers’ growth performance (feed conversion ratio, kg/kg).
Thermoneutral temperature.
Heat stress temperature.
Thermoneutral environment, control feed.
Heat stress environment, control feed.
Heat stress environment and specialty feed supplement fortified diet.
% mitigation = (HS treatment - HS control)/(TN control – HS control) x 100.
HS environment 4–10 hours per day in the range of three days per week to daily up to 10 days.
Continuous HS environment (24 hours/day) usually during the second half of the fattening.
Lactobacillus plantarum, Lactobacillus delbrueckii ssp. Bulgaricus, Lactobacillus acidophilus, Lactobacillus rhamnosus, Bifidobacterium bifidum, and Streptococcus salivarius ssp. Thermophilus and Enterococcus faecium.
Lactobacillus pentosus ITA23 and Lactobacillus acidophilus ITA44.
Saccharomyces cerevisiae and Lactobacillus acidophilus.
Combination of two or more feed additives are used.
Only one feed additive used.
HS type averages regardless of single or combined supplementation.
It can be seen that - overall - the most difficult parameter to improve is that of feed intake (Table 7), as the feed additives studied have quite variable mitigation capacities. The overall mitigation percentage is only about 5–15%.Bettervalueswere obtained in the case of chronic heat stress compared to cyclic heat stress, but different feed additives were used. The highest improvement (above 85%) was achieved with a probiotic [88]. However, other probiotics were much less effective.
The adversely affected daily gain can be improved by about 30–35%, and the feed conversion ratio increased up to 60–70%. Observing the mitigation capacity of different feed additives in this regard, it seems that probiotics and vitamins can be the most effective mitigators, especially when they are applied in combination [84]. However, further research is needed to determine the most effective microbe combination(s), and the most effective levels of vitamins, as well as their interactive effects. Only one study reported results with fumaric acid supplementation which also seems promising, but more research is still needed [86]. Quite a few authors have tested the feed supplements in combination, which is in line with feed industry trends. Therefore, we calculated the average % mitigation value for combined and single applications. Data shows that combined applications are more effective in cyclic heat stress conditions, while that benefit cannot be observed in chronic heat stress.
7. Conclusions
Based on the scientific findings presented in this chapter, the following important conclusions can be drawn:
Using fat in the diet (up to 5%) can reduce heat production in livestock.
Vitamins (e.g. A, E and C) are capable of reacting with free radicals, thereby reducing their amounts and lipid peroxidation in the poultry. However, micro minerals (e.g. Zn, Se) are not directly capable of preventing or reducing ROS-formation, but they are essential cofactors for those enzymes which are reacting with free radicals.
Vitamin E and Vitamin C supplementation improved antioxidant parameters (CAT, GR, GSH, MDA, SOD) due to their essential antioxidant function. Both Zn and Se are also improving antioxidant parameters (GR, GSH, GPx, and MDA).
Antioxidant potential of vitamins and micro minerals is more efficient in combination under heat stress in poultry nutrition.
Plant extracts (e.g. oregano) could decrease the negative effects of heat stress on antioxidant enzyme activity due its antioxidant constituents.
Betaine reduces heat production in animals at high ambient temperatures.
Acute heat stress induced a drop in feed intake and increased nutrient demand will result even in weight loss. However, if heat stress is prolonged, adaptation will occur.
Probiotics and vitamins (C and E) seem to be the most effective means of reducing the negative effects of heat stress.
Main conclusion for the practice: Different feed additives and supplementation strategies (single vs. combined) can be more effective in temperate and tropical countries. Therefore, in order to decide which feed additive to use and in what form (single or combined) the most effectively, it is recommended that farmers carry out a pre-study in the given climatic and feeding conditions.
\n',keywords:"Broiler, Feed additives, Heat stress, Antioxidant status, Performance",chapterPDFUrl:"https://cdn.intechopen.com/pdfs/79279.pdf",chapterXML:"https://mts.intechopen.com/source/xml/79279.xml",downloadPdfUrl:"/chapter/pdf-download/79279",previewPdfUrl:"/chapter/pdf-preview/79279",totalDownloads:248,totalViews:0,totalCrossrefCites:0,dateSubmitted:null,dateReviewed:"October 1st 2021",datePrePublished:"November 15th 2021",datePublished:"December 8th 2021",dateFinished:"November 10th 2021",readingETA:"0",abstract:"Global warming is one of the major challenges for mankind, with animal breeding one of the most affected sectors in the agricultural industry. High ambient temperatures negatively affect all domestic animals. While it is true that pork and dairy production suffer the consequences of heat waves, it is actually the poultry industry which is hit the hardest by the heat stress poultry must endure due to hotter weather. Consequently, we have a fundamental interest in reducing and/or eliminating the negative effects of climate change, i.e. prolonged high ambient temperatures. The aim of this chapter is to present the adverse effects of heat stress on energy metabolism, anti- and pro-oxidant capacity and production in birds. A further goal is to show how various feed additives (e.g. vitamin A, C and E, selenium, zinc, betaine, plant extract, and probiotics) can reduce the negative effects of heat stress. Based on the large number of recent scientific findings, the following conclusions were drawn: Using fat in the diet (up to 5%) can reduce heat production in livestock. Vitamins (e.g. A, E and C) are capable of reacting with free radicals. Vitamin E and Vitamin C, Zn, and Se supplementation improved antioxidant parameters. Antioxidant potential of vitamins and micro minerals is more efficient in combination under heat stress in poultry nutrition. Plant extracts (e.g. oregano) could decrease the negative effects of heat stress on antioxidant enzyme activity due to its antioxidant constituents. Betaine reduces heat production in animals at high ambient temperatures. While acute heat stress induces a drop in feed intake, with the resulting increased nutrient demand leading to weight loss, if heat stress is prolonged, adaptation will occur. Probiotics and vitamins (C and E) seem to be the most effective means to reduce the negative effects of heat stress.",reviewType:"peer-reviewed",bibtexUrl:"/chapter/bibtex/79279",risUrl:"/chapter/ris/79279",signatures:"László Babinszky, Csaba Szabó and Márta Horváth",book:{id:"10496",type:"book",title:"Advanced Studies in the 21st Century Animal Nutrition",subtitle:null,fullTitle:"Advanced Studies in the 21st Century Animal Nutrition",slug:"advanced-studies-in-the-21st-century-animal-nutrition",publishedDate:"December 8th 2021",bookSignature:"László Babinszky, Juliana Oliveira and Edson Mauro Santos",coverURL:"https://cdn.intechopen.com/books/images_new/10496.jpg",licenceType:"CC BY 3.0",editedByType:"Edited by",isbn:"978-1-83969-404-2",printIsbn:"978-1-83969-403-5",pdfIsbn:"978-1-83969-405-9",isAvailableForWebshopOrdering:!0,editors:[{id:"53998",title:"Prof.",name:"László",middleName:null,surname:"Babinszky",slug:"laszlo-babinszky",fullName:"László Babinszky"}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"}},authors:[{id:"53998",title:"Prof.",name:"László",middleName:null,surname:"Babinszky",fullName:"László Babinszky",slug:"laszlo-babinszky",email:"babinszky@agr.unideb.hu",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/53998/images/system/53998.png",institution:{name:"University of Debrecen",institutionURL:null,country:{name:"Hungary"}}},{id:"345713",title:"Dr.",name:"Csaba",middleName:null,surname:"Szabó",fullName:"Csaba Szabó",slug:"csaba-szabo",email:"szabo.csaba@agr.unideb.hu",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:{name:"University of Debrecen",institutionURL:null,country:{name:"Hungary"}}},{id:"345719",title:"Mrs.",name:"Márta",middleName:null,surname:"Horváth",fullName:"Márta Horváth",slug:"marta-horvath",email:"mhorvath@agr.unideb.hu",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",institution:{name:"University of Debrecen",institutionURL:null,country:{name:"Hungary"}}}],sections:[{id:"sec_1",title:"1. Introduction",level:"1"},{id:"sec_2",title:"2. Methodology of the literature review",level:"1"},{id:"sec_3",title:"3. Heat production of animals and heat stress",level:"1"},{id:"sec_4",title:"4. Reduction of heat production by nutritional tools",level:"1"},{id:"sec_4_2",title:"4.1 Using fat in the diets",level:"2"},{id:"sec_5_2",title:"4.2 Using vitamin C in chicken diet to change energy metabolism",level:"2"},{id:"sec_6_2",title:"4.3 Using betaine as feed additive in the diet",level:"2"},{id:"sec_8",title:"5. Effects of heat stress on anti- and prooxidant status in birds. Mitigation using different feed additives",level:"1"},{id:"sec_8_2",title:"5.1 Anti- and prooxidant status",level:"2"},{id:"sec_8_3",title:"5.1.1 Impacts of heat stress",level:"3"},{id:"sec_9_3",title:"Table 2.",level:"3"},{id:"sec_9_4",title:"Table 2.",level:"4"},{id:"sec_9_5",title:"Table 2.",level:"5"},{id:"sec_10_5",title:"5.1.2.1.2 Vitamin C",level:"5"},{id:"sec_12_4",title:"Table 3.",level:"4"},{id:"sec_12_5",title:"Table 3.",level:"5"},{id:"sec_13_5",title:"Table 4.",level:"5"},{id:"sec_15_4",title:"5.1.2.3 Plant extracts",level:"4"},{id:"sec_16_4",title:"5.1.2.4 Probiotics",level:"4"},{id:"sec_20",title:"6. The effect of heat stress on the performance of broilers reduced by using feed additives",level:"1"},{id:"sec_20_2",title:"6.1 The effect of heat stress on the performance of broilers",level:"2"},{id:"sec_21_2",title:"6.2 Mitigation capacity of various feed additives on HS in broilers",level:"2"},{id:"sec_23",title:"7. 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Spanish Journal of Agricultural Research. 16(2):e0505. DOI: 10.5424/sjar/2018162-12753'},{id:"B90",body:'Ma, B., X. He, Z. Lu, L. Zhang, J. Li, Y. Jiang, G. Zhou, F. Gao. 2018. Chronic heat stress affects muscle hypertrophy, muscle protein synthesis and uptake of amino acid in broilers via insulin like growth factor-mammalian target of rapamycin signal pathway. Poultry Science. 97:4150–4158. DOI: 10.3382/ps/pey291'},{id:"B91",body:'Goo, D., J. H. Kim, G. Park, J. Reyes, D. Kil. 2019. Effect of heat stress and stocking density on growth performance, Breast meat quality, and intestinal barrier function in broiler chickens. Animals. 9(3):107. DOI: 10.3390/ani9030107'},{id:"B92",body:'Goel, A. 2021. Heat stress management in poultry. Journal of Animal Physiology and Animal Nutrition. 00:1–10. DOI: 10.1111/jpn.13496'},{id:"B93",body:'Ortega, A. D. S. V., C. Szabó. 2021. Adverse effects of heat stress on the intestinal integrity and function of pigs and the mitigation capacity of dietary antioxidants: a review. Animals. 11(4):1135. DOI: 10.3390/ani11041135'}],footnotes:[],contributors:[{corresp:"yes",contributorFullName:"László Babinszky",address:"babinszky@agr.unideb.hu",affiliation:'
Department of Animal Nutrition and Physiology, University of Debrecen, Debrecen, Hungary
Department of Animal Nutrition and Physiology, University of Debrecen, Debrecen, Hungary
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Munthali, Richard M. Mkandawire and Nasson Tembo",authors:[{id:"76161",title:"Dr.",name:"Simon Muchina",surname:"Munthali",fullName:"Simon Muchina Munthali",slug:"simon-muchina-munthali",email:"Muchina.munthali@gmail.com"}],book:{id:"890",title:"Rural Development",slug:"rural-development-contemporary-issues-and-practices",productType:{id:"1",title:"Edited Volume"}}},{id:"56778",title:"Integration of Ecological and Socioeconomic Factors in Securing Wildlife Dispersal Corridors in the Kavango-Zambezi Transfrontier Conservation Area, Southern Africa",slug:"integration-of-ecological-and-socioeconomic-factors-in-securing-wildlife-dispersal-corridors-in-the-",abstract:"Transfrontier conservation areas (TFCAs) are being established throughout southern Africa to integrating biodiversity conservation and rural development at the transboundary landscape scale. Among the nine TFCAs that have been established over the past 20 years, the Kavango-Zambezi (KAZA) TFCA) is the most grandiose in terms of size (≈ 520,000 Km2), number of partner countries involved (five), elephant (Loxodonta africana) population (≈ 199,031, which is the largest on the African continent), and encompasses 36 protected areas of various categories, interspaced by communal and private lands. The TFCA concept aims to ensure that key ecological processes continue to function where borders have divided ecosystems, and wildlife migration corridors. Attainment of this ecological objective is however being constrained by the anthropogenic threats, mostly poaching, and habitat fragmentation. These threats are being aggravated by the increasing human population, climate variability and underdeveloped rural livelihoods. To restore ecological processes, the following tactics have been recommended: (a) strengthening of transboundary law enforcement to effectively reduce poaching, and illegal offtake of timber; (b) establishment of “Stepping Stones” in the form of conservancies and fishing protected zones at wildlife crossing point on the major river systems; (c) reducing dependence on wood-fuel, and ensuring sustainable provision of affordable and reliable modern sources of energy; (d) adoption of the commodity-based trade standards in the production of beef for the export market to reduce the impact of veterinary fences on the dispersing wildlife; (e) implementation of early-season burning around all the sensitive biomes to protect them from the destructive late dry season fires; (f) adoption of conservation agriculture as a tool for improving land husbandry, intensification of agriculture, and decreasing the likelihood of cutting down forested areas to plant new agriculture fields; and (g) reducing the impact of climate variability on wildlife by providing artificial water – guided by environmental impact assessments. To enhance the socioeconomic development of the local communities and win them as allies in securing the wildlife dispersal corridors, the following actions should be adopted: (a) promotion of community-private partnerships in ecotourism development – alongside the establishment of a revolving loan fund to enable local communities’ access flexible source of capital for investment in ecotourism and auxiliary business opportunities; (b) promotion of biodiversity stewardship as an incentive for the local communities to commit their land to the sustenance of the wildlife dispersal corridors; (c) reducing human wildlife conflicts, through macro, meso and micro-level land-use planning to spatially delineate land committed to various categories, including protected areas, wildlife dispersal areas, and developed and communal areas; and (d) promotion of harmonised enabling policies and legislation to facilitate slowing down of human population growth, which is one of the prime triggers of habitat fragmentation in the KAZA TFCA.",signatures:"Simon M. 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He also has Certificates in Conflict Analysis and Negotiation and Conflict Management from the United States Institute of Peace. Dr. Adisa has, since obtaining his Bachelor’s degree in 1989, been active in AE&RD activities within and outside Nigeria. His research works have been published extensively in scholarly journals within and outside Africa, and he is on the Editorial/Reviewers’ Boards of several scholarly professional journals. Dr. Adisa is an active member of notable professional associations, including the Association for International Agricultural and Extension Education (AIAEE).",institutionString:null,institution:{name:"University of Ilorin",institutionURL:null,country:{name:"Nigeria"}}},{id:"77634",title:"Dr.",name:"Miguel A.",surname:"Damian-Huato",slug:"miguel-a.-damian-huato",fullName:"Miguel A. Damian-Huato",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Benemérita Universidad Autónoma de Puebla",institutionURL:null,country:{name:"Mexico"}}},{id:"79134",title:"Dr.",name:"Iker",surname:"Etxano",slug:"iker-etxano",fullName:"Iker Etxano",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/79134/images/4532_n.jpg",biography:null,institutionString:null,institution:{name:"University of the Basque Country",institutionURL:null,country:{name:"Spain"}}},{id:"81811",title:"MSc.",name:"Michel",surname:"Dulcire",slug:"michel-dulcire",fullName:"Michel Dulcire",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Centre de Coopération Internationale en Recherche Agronomique pour le Développement",institutionURL:null,country:{name:"France"}}},{id:"82709",title:"Dr.",name:"Ravinder",surname:"Rena",slug:"ravinder-rena",fullName:"Ravinder Rena",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/82709/images/system/82709.jpg",biography:"Professor, Dr Ravinder Rena is a profound academician and distinguished scholar in economics, writer, editor and adviser with over 28 years of teaching and research experience in Asia-Pacific, Africa and European continents. He has been an independent consultant and training expert. Prof Rena is currently working as Professor of Economics and Internationalization Project Leader at the NWU Business School, North-West University, South Africa. He also serves as an Adjunct Professor of Economics for Master’s and Doctorate Programmes at Monarch Business School, Monarch University, Hagendorn, Zug, Switzerland. Previously, he worked as a Professor of Economics at the Department of Economics, University of the Western Cape, Cape Town, South Africa. He also served as Professor and the Coordinator, Joint African Master's Programme (JAMP), Harold Pupkewitz Graduate School of Business at the Namibia University of Science and Technology (NUST). He also worked as the Head of Economics, Department of Business Studies, the Papua New Guinea University of Technology, Papua New Guinea. Prof. Rena earned his Ph.D. from the Department of Economics, Osmania University, Hyderabad, India, with an achievement of University Gold Medal (for the Best Ph.D. candidates/ thesis in the field of Economics/ Commerce/ Business Management). He obtained his M.A., B.Ed., M.Phil., Ph.D from Osmania University and B.A., and LL.B., from Kakatiya University, Warangal, India. \r\nHe got the prestigious Award of the Best Researcher 2011 from Namibia University of Science and Technology and also the recipient of the esteemed Award -Man of the Year 2012 which was bestowed upon him by the American Biographic Institute (ABI), USA. Based on his extraordinary research contributions, American Historical Society, USA has been enlisted his profile in the World Who is Who International. He serves as the Chairman of the Panel of Judges for Economics of the Global UnderGraduate (UA) Award based in Dublin, Ireland, United Kingdom. He wrote 10 books and published more than 120 articles in reputed National and International peer-reviewed Journals. Besides, he has published more than 150 articles in Newspapers and Magazines and Websites and presented over 40 papers in the national and international conferences and seminars. He is the founding Editor-In-Chief of an International Journal of Education Economics and Development (IJEED) and Associate Editor of African Journal of Science, Technology, Innovation and Development (AJSTID) which are IBSS and Scopus indexed Journals published by Inderscience, UK and Taylor and Francis UK respectively. He serves as an Editorial Board Member for many reputed Journals. He has been a Keynote Speaker for various national and international conferences in Europe, America, Africa and Asian continents. As of December 2019, his author Rank is 5,399 out of 438,702 of Social Science Research Network (SSRN),USA in which he is among the top 1.3% researchers in the world. \r\nHe has widely written in the areas of Economics of Education, Development Economics and Inclusive growth, Globalization and Higher Education, Microfinance, Rural Entrepreneurship, WTO, Sustainable development, FDIs, development aid in Africa. He supervised Ph.Ds, M.Phils and Masters’ theses and dissertations across the globe. He served many academic and professional committees as a Chairman and as a Panel Member. He is also been an external examiner/moderator for Masters and Ph.D of different Universities of Africa, Europe, Asia, America and Australia. He traveled to over 40 countries in the world.",institutionString:"NWU Business School",institution:null},{id:"87343",title:"Dr.",name:"Artemio",surname:"Cruz Leon",slug:"artemio-cruz-leon",fullName:"Artemio Cruz Leon",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Chapingo Autonomous University",institutionURL:null,country:{name:"Mexico"}}},{id:"87345",title:"Dr.",name:"Benito",surname:"Ramirez Valverde",slug:"benito-ramirez-valverde",fullName:"Benito Ramirez Valverde",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:null},{id:"87348",title:"Dr.",name:"Agustin",surname:"Aragon Garcia",slug:"agustin-aragon-garcia",fullName:"Agustin Aragon Garcia",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Benemérita Universidad Autónoma de Puebla",institutionURL:null,country:{name:"Mexico"}}},{id:"116577",title:"Dr.",name:"Chia",surname:"Eduardo",slug:"chia-eduardo",fullName:"Chia Eduardo",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Institut National de la Recherche Agronomique",institutionURL:null,country:{name:"Morocco"}}},{id:"116578",title:"Dr.",name:"Piraux",surname:"Marc",slug:"piraux-marc",fullName:"Piraux Marc",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Centre de Coopération Internationale en Recherche Agronomique pour le Développement",institutionURL:null,country:{name:"France"}}}]},generic:{page:{slug:"why-publish-with-intechopen",title:"Why publish with IntechOpen?",intro:"
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He has contributed in stochastic estimation of control area especially, in the Multiple Target Tracking and Interactive Multiple Model (IMM) research, Ball & Beam Control Problem, Robotics, Levitation Control. He has contributed in developing Algorithms for Fingerprint Matching, Computer Vision and Face Recognition. He has been supervising Pattern Recognition, Formal Languages and Distributed Processing projects for several years. He has reviewed many books on Management, Computer Science. Currently, he is an active and permanent reviewer for many international conferences and symposia and the program committee member for many international conferences.\nIn teaching he has taught the core computer science subjects like, Digital Design, Real Time Embedded System Programming, Operating Systems, Software Engineering, Data Structures, Databases, Compiler Construction. 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Over the past few decades, no major new types of antibiotics have been produced and almost all known antibiotics are increasingly losing their activity against pathogenic microorganisms. The levels of multi-drug resistant bacteria have also increased. It is known that worldwide, more than 60% of all antibiotics that are produced find their use in animal production for both therapeutic and non-therapeutic purposes. The use of antimicrobial agents in animal husbandry has been linked to the development and spread of resistant bacteria. Poultry products are among the highest consumed products worldwide but a lot of essential antibiotics are employed during poultry production in several countries; threatening the safety of such products (through antimicrobial residues) and the increased possibility of development and spread of microbial resistance in poultry settings. This chapter documents some of the studies on antibiotic usage in poultry farming; with specific focus on some selected bacterial species, their economic importance to poultry farming and reports of resistances of isolated species from poultry settings (farms and poultry products) to essential antibiotics.",book:{id:"6978",slug:"antimicrobial-resistance-a-global-threat",title:"Antimicrobial Resistance",fullTitle:"Antimicrobial Resistance - A Global Threat"},signatures:"Christian Agyare, Vivian Etsiapa Boamah, Crystal Ngofi Zumbi and\nFrank Boateng Osei",authors:[{id:"182058",title:"Dr.",name:"Christian",middleName:null,surname:"Agyare",slug:"christian-agyare",fullName:"Christian Agyare"},{id:"261271",title:"MSc.",name:"Crystal Ngofi",middleName:null,surname:"Zumbi",slug:"crystal-ngofi-zumbi",fullName:"Crystal Ngofi Zumbi"},{id:"261272",title:"MSc.",name:"Frank Boateng",middleName:null,surname:"Osei",slug:"frank-boateng-osei",fullName:"Frank Boateng Osei"},{id:"261273",title:"Dr.",name:"Vivian Etsiapa",middleName:null,surname:"Boamah",slug:"vivian-etsiapa-boamah",fullName:"Vivian Etsiapa Boamah"}]},{id:"39599",doi:"10.5772/50046",title:"Encapsulation Technology to Protect Probiotic Bacteria",slug:"encapsulation-technology-to-protect-probiotic-bacteria",totalDownloads:12297,totalCrossrefCites:40,totalDimensionsCites:81,abstract:null,book:{id:"3145",slug:"probiotics",title:"Probiotics",fullTitle:"Probiotics"},signatures:"María Chávarri, Izaskun Marañón and María Carmen Villarán",authors:[{id:"150285",title:"Dr.",name:"María",middleName:null,surname:"Chávarri Hueda",slug:"maria-chavarri-hueda",fullName:"María Chávarri Hueda"},{id:"151613",title:"MSc.",name:"Izaskun",middleName:null,surname:"Marañon",slug:"izaskun-maranon",fullName:"Izaskun Marañon"},{id:"151621",title:"Dr.",name:"Mª Carmen",middleName:null,surname:"Villarán",slug:"ma-carmen-villaran",fullName:"Mª Carmen Villarán"}]},{id:"39607",doi:"10.5772/50121",title:"Recent Application of Probiotics in Food and Agricultural Science",slug:"recent-application-of-probiotics-in-food-and-agricultural-science",totalDownloads:10098,totalCrossrefCites:28,totalDimensionsCites:74,abstract:null,book:{id:"3145",slug:"probiotics",title:"Probiotics",fullTitle:"Probiotics"},signatures:"Danfeng Song, Salam Ibrahim and Saeed Hayek",authors:[{id:"107905",title:"Prof.",name:"Salam",middleName:null,surname:"Ibrahim",slug:"salam-ibrahim",fullName:"Salam Ibrahim"},{id:"150202",title:"Dr.",name:"Danfeng",middleName:null,surname:"Song",slug:"danfeng-song",fullName:"Danfeng Song"},{id:"151025",title:"MSc.",name:"Saeed",middleName:null,surname:"Hayek",slug:"saeed-hayek",fullName:"Saeed Hayek"}]},{id:"49246",doi:"10.5772/61300",title:"Chitosan as a Biomaterial — Structure, Properties, and Electrospun Nanofibers",slug:"chitosan-as-a-biomaterial-structure-properties-and-electrospun-nanofibers",totalDownloads:4607,totalCrossrefCites:24,totalDimensionsCites:55,abstract:"Chitosan is a polysaccharide derived from chitin; chitin is the second most abundant polysaccharide in the world, after cellulose. Chitosan is biocompatible, biodegradable and non-toxic, so that it can be usedin medicalapplications such as antimicrobial and wound healing biomaterials. It also used as chelating agent due to its ability to bind with cholesterol, fats, proteins and metal ions.",book:{id:"4648",slug:"concepts-compounds-and-the-alternatives-of-antibacterials",title:"Concepts, Compounds and the Alternatives of Antibacterials",fullTitle:"Concepts, Compounds and the Alternatives of Antibacterials"},signatures:"H. M. Ibrahim and E.M.R. El- Zairy",authors:[{id:"90645",title:"Dr.",name:"Hassan",middleName:null,surname:"Ibrahim",slug:"hassan-ibrahim",fullName:"Hassan Ibrahim"},{id:"175694",title:"Dr.",name:"Enas",middleName:null,surname:"El- Zairy",slug:"enas-el-zairy",fullName:"Enas El- Zairy"}]},{id:"51065",doi:"10.5772/63499",title:"Role of the Biofilms in Wastewater Treatment",slug:"role-of-the-biofilms-in-wastewater-treatment",totalDownloads:6759,totalCrossrefCites:24,totalDimensionsCites:55,abstract:"Biological wastewater treatment systems play an important role in improving water quality and human health. This chapter thus briefly discusses different biological methods, specially biofilm technologies, the development of biofilms on different filter media, factors affecting their development as well as their structure and function. It also tackles various conventional and modern molecular techniques for detailed exploration of the composition, diversity and dynamics of biofilms. These data are crucial to improve the performance, robustness and stability of biofilm-based wastewater treatment technologies.",book:{id:"5197",slug:"microbial-biofilms-importance-and-applications",title:"Microbial Biofilms",fullTitle:"Microbial Biofilms - Importance and Applications"},signatures:"Shama Sehar and Iffat Naz",authors:[{id:"180364",title:"Dr.",name:"Iffat",middleName:null,surname:"Naz",slug:"iffat-naz",fullName:"Iffat Naz"},{id:"183345",title:"Dr.",name:"Shama",middleName:null,surname:"Sehar",slug:"shama-sehar",fullName:"Shama Sehar"}]}],mostDownloadedChaptersLast30Days:[{id:"65613",title:"The Methods for Detection of Biofilm and Screening Antibiofilm Activity of Agents",slug:"the-methods-for-detection-of-biofilm-and-screening-antibiofilm-activity-of-agents",totalDownloads:9033,totalCrossrefCites:11,totalDimensionsCites:20,abstract:"Biofilm producer microorganisms cause nosocomial and recurrent infections. Biofilm that is a sticky exopolysaccharide is the main virulence factor causing biofilm-related infections. Biofilm formation begins with attachment of bacteria to biotic surface such as host cell or abiotic surface such as prosthetic devices. After attachment, aggregation of bacteria is started by cell-cell adhesion. Aggregation continues with the maturation of biofilm. Dispersion is started by certain conditions such as phenol-soluble modulins (PSMs). By this way, sessile bacteria turn back into planktonic form. Bacteria embedded in biofilm (sessile form) are more resistant to antimicrobials than planktonic bacteria. So it is hard to treat biofilm-embedded bacteria than planktonic forms. For this reason, it is important to detect biofilm. There are a few biofilm detection and biofilm production methods on prosthetics, methods for screening antibacterial effect of agents against biofilm-embedded microorganism and antibiofilm effect of agents against biofilm production and mature biofilm. The aim of this chapter is to overview direct and indirect methods such as microscopy, fluorescent in situ hybridization, and Congo red agar, tube method, microtiter plate assay, checkerboard assay, plate counting, polymerase chain reaction, mass spectrometry, MALDI-TOF, and biological assays used by antibiofilm researches.",book:{id:"8427",slug:"antimicrobials-antibiotic-resistance-antibiofilm-strategies-and-activity-methods",title:"Antimicrobials, Antibiotic Resistance, Antibiofilm Strategies and Activity Methods",fullTitle:"Antimicrobials, Antibiotic Resistance, Antibiofilm Strategies and Activity Methods"},signatures:"Sahra Kırmusaoğlu",authors:[{id:"179460",title:"Associate Prof.",name:"Sahra",middleName:null,surname:"Kırmusaoğlu",slug:"sahra-kirmusaoglu",fullName:"Sahra Kırmusaoğlu"}]},{id:"62553",title:"Antibiotic Use in Poultry Production and Its Effects on Bacterial Resistance",slug:"antibiotic-use-in-poultry-production-and-its-effects-on-bacterial-resistance",totalDownloads:7129,totalCrossrefCites:42,totalDimensionsCites:82,abstract:"A surge in the development and spread of antibiotic resistance has become a major cause for concern. Over the past few decades, no major new types of antibiotics have been produced and almost all known antibiotics are increasingly losing their activity against pathogenic microorganisms. The levels of multi-drug resistant bacteria have also increased. It is known that worldwide, more than 60% of all antibiotics that are produced find their use in animal production for both therapeutic and non-therapeutic purposes. The use of antimicrobial agents in animal husbandry has been linked to the development and spread of resistant bacteria. Poultry products are among the highest consumed products worldwide but a lot of essential antibiotics are employed during poultry production in several countries; threatening the safety of such products (through antimicrobial residues) and the increased possibility of development and spread of microbial resistance in poultry settings. This chapter documents some of the studies on antibiotic usage in poultry farming; with specific focus on some selected bacterial species, their economic importance to poultry farming and reports of resistances of isolated species from poultry settings (farms and poultry products) to essential antibiotics.",book:{id:"6978",slug:"antimicrobial-resistance-a-global-threat",title:"Antimicrobial Resistance",fullTitle:"Antimicrobial Resistance - A Global Threat"},signatures:"Christian Agyare, Vivian Etsiapa Boamah, Crystal Ngofi Zumbi and\nFrank Boateng Osei",authors:[{id:"182058",title:"Dr.",name:"Christian",middleName:null,surname:"Agyare",slug:"christian-agyare",fullName:"Christian Agyare"},{id:"261271",title:"MSc.",name:"Crystal Ngofi",middleName:null,surname:"Zumbi",slug:"crystal-ngofi-zumbi",fullName:"Crystal Ngofi Zumbi"},{id:"261272",title:"MSc.",name:"Frank Boateng",middleName:null,surname:"Osei",slug:"frank-boateng-osei",fullName:"Frank Boateng Osei"},{id:"261273",title:"Dr.",name:"Vivian Etsiapa",middleName:null,surname:"Boamah",slug:"vivian-etsiapa-boamah",fullName:"Vivian Etsiapa Boamah"}]},{id:"65914",title:"Introductory Chapter: The Action Mechanisms of Antibiotics and Antibiotic Resistance",slug:"introductory-chapter-the-action-mechanisms-of-antibiotics-and-antibiotic-resistance",totalDownloads:4310,totalCrossrefCites:6,totalDimensionsCites:8,abstract:null,book:{id:"8427",slug:"antimicrobials-antibiotic-resistance-antibiofilm-strategies-and-activity-methods",title:"Antimicrobials, Antibiotic Resistance, Antibiofilm Strategies and Activity Methods",fullTitle:"Antimicrobials, Antibiotic Resistance, Antibiofilm Strategies and Activity Methods"},signatures:"Sahra Kırmusaoğlu, Nesrin Gareayaghi and Bekir S. Kocazeybek",authors:[{id:"179460",title:"Associate Prof.",name:"Sahra",middleName:null,surname:"Kırmusaoğlu",slug:"sahra-kirmusaoglu",fullName:"Sahra Kırmusaoğlu"},{id:"248288",title:"Prof.",name:"Bekir",middleName:null,surname:"Kocazeybek",slug:"bekir-kocazeybek",fullName:"Bekir Kocazeybek"},{id:"406463",title:"Dr.",name:"Nesrin",middleName:null,surname:"Gareayaghi",slug:"nesrin-gareayaghi",fullName:"Nesrin Gareayaghi"}]},{id:"50992",title:"Probiotics: A Comprehensive Review of Their Classification, Mode of Action and Role in Human Nutrition",slug:"probiotics-a-comprehensive-review-of-their-classification-mode-of-action-and-role-in-human-nutrition",totalDownloads:5325,totalCrossrefCites:14,totalDimensionsCites:26,abstract:"Probiotics are live microorganisms that live in gastrointestinal (GI) tract and are beneficial for their hosts and prevent certain diseases. In this chapter, after a complete introduction to probiotics, definition, mechanism of action, and their classification, currently used organisms will be discussed in detail. Moreover, different kinds of nutritional synthetic products of probiotics along with their safety and drug interaction will be noticed. This chapter mentions all clinical trial studies that have been done to evaluate probiotic efficacy with a focus on gastrointestinal diseases.",book:{id:"5193",slug:"probiotics-and-prebiotics-in-human-nutrition-and-health",title:"Probiotics and Prebiotics in Human Nutrition and Health",fullTitle:"Probiotics and Prebiotics in Human Nutrition and Health"},signatures:"Amirreza Khalighi, Reza Behdani and Shabnam Kouhestani",authors:[{id:"179560",title:"Dr.",name:"Amirreza",middleName:null,surname:"Khalighi",slug:"amirreza-khalighi",fullName:"Amirreza Khalighi"},{id:"185238",title:"Dr.",name:"Reza",middleName:null,surname:"Behdani",slug:"reza-behdani",fullName:"Reza Behdani"},{id:"185239",title:"Dr.",name:"Shabnam",middleName:null,surname:"Kouhestani",slug:"shabnam-kouhestani",fullName:"Shabnam Kouhestani"}]},{id:"56849",title:"Physiology and Pathology of Innate Immune Response Against Pathogens",slug:"physiology-and-pathology-of-innate-immune-response-against-pathogens",totalDownloads:6083,totalCrossrefCites:20,totalDimensionsCites:26,abstract:"Pathogen infections are recognized by the immune system, which consists of two types of responses: an innate immune response and an antigen-specific adaptive immune response. The innate response is characterized by being the first line of defense that occurs rapidly in which leukocytes such as neutrophils, monocytes, macrophages, eosinophils, mast cells, dendritic cells, etc., are involved. These cells recognize the pathogen-associated molecular patterns (PAMPs), which have been evolutionarily conserved by the diversity of microorganisms that infect humans. Recognition of these pathogen-associated molecular patterns occurs through pattern recognition receptors such as Toll-like receptors and some other intracellular receptors such as nucleotide oligomerization domain (NOD), with the aim of amplifying the inflammation and activating the adaptive cellular immune response, through the antigenic presentation. In the present chapter, we will review the importance of the main components involved in the innate immune response, such as different cell types, inflammatory response, soluble immune mediators and effector mechanisms exerted by the immune response against bacteria, viruses, fungi, and parasites; all with the purpose of eliminating them and eradicating the infection of the host.",book:{id:"5975",slug:"physiology-and-pathology-of-immunology",title:"Physiology and Pathology of Immunology",fullTitle:"Physiology and Pathology of Immunology"},signatures:"José Luis Muñoz Carrillo, Flor Pamela Castro García, Oscar\nGutiérrez Coronado, María Alejandra Moreno García and Juan\nFrancisco Contreras Cordero",authors:[{id:"214236",title:"Dr.",name:"Jose Luis",middleName:null,surname:"Muñoz-Carrillo",slug:"jose-luis-munoz-carrillo",fullName:"Jose Luis Muñoz-Carrillo"},{id:"216080",title:"Dr.",name:"Alejandra",middleName:null,surname:"Moreno-García",slug:"alejandra-moreno-garcia",fullName:"Alejandra Moreno-García"},{id:"216081",title:"Dr.",name:"Oscar",middleName:null,surname:"Gutiérrez-Coronado",slug:"oscar-gutierrez-coronado",fullName:"Oscar Gutiérrez-Coronado"},{id:"216082",title:"Dr.",name:"Pamela",middleName:null,surname:"Castro-García",slug:"pamela-castro-garcia",fullName:"Pamela Castro-García"},{id:"220717",title:"Dr.",name:"Juan Francisco",middleName:null,surname:"Contreras Cordero",slug:"juan-francisco-contreras-cordero",fullName:"Juan Francisco Contreras Cordero"}]}],onlineFirstChaptersFilter:{topicId:"13",limit:6,offset:0},onlineFirstChaptersCollection:[{id:"81760",title:"On the Selective Isolation of Actinobacteria from Different Mexican Ecosystems",slug:"on-the-selective-isolation-of-actinobacteria-from-different-mexican-ecosystems",totalDownloads:6,totalDimensionsCites:0,doi:"10.5772/intechopen.104699",abstract:"Actinobacteria isolated from less studied sites on our planet represent a huge opportunity for the discovery of novel microorganisms that may produce unique compounds with biological activity. The class actinobacteria encompasses 80% of the microbes that produce the antibacterial compounds used in medicine today. However, the resistance acquired/showed by pathogenic microorganisms opens the opportunity to explore Mexican ecosystems as a source of novel actinobacteria. Air samples have shown to be an excellent site of study, marine ecosystems which include sediments and marine organisms are important sources of novel actinobacteria and soil samples are still a promising source to isolate this microbial group. The isolation of novel actinobacteria is a dynamic strategy that depends on the expertise, patience, and talent of the techniques applied and needs to be fully explored to untap the unknown actinobacterial diversity with potential in biology.",book:{id:"10893",title:"Actinobacteria",coverURL:"https://cdn.intechopen.com/books/images_new/10893.jpg"},signatures:"Erika T. Quintana, Luis A. Maldonado, Luis Contreras-Castro, Amanda Alejo-Viderique, Martha E. Esteva García, Claudia J. Hernández-Guerrero, Juan C. Cancino-Díaz, Carlos Sánchez, Luis A. Ladino, Juan Esteban Martínez-Gómez and Noemí Matías-Ferrer"},{id:"81741",title:"Chronic Intraocular Leptospiral Infection Relying on Biofilm Formation inside the Vitreous Cavity Leads to Recurrent Uveitis in Horses",slug:"chronic-intraocular-leptospiral-infection-relying-on-biofilm-formation-inside-the-vitreous-cavity-le",totalDownloads:2,totalDimensionsCites:0,doi:"10.5772/intechopen.104527",abstract:"Equine recurrent uveitis (ERU) is a disease known and feared for centuries, as it almost always leads to blindness even with careful and meticulous conservative treatment of the individual episodes of uveitis. In about one-third of horses, both eyes are affected, often necessitating euthanasia. A link between ERU and leptospiral infection has been suspected for nearly 80 years. Vitreous lavage (vitrectomy) can preserve vision in affected eyes. After surgery, no further episodes of uveitis occur in up to more than 95% of operated eyes. With routine performance of vitrectomies, numerous vitreous samples could be used for further investigations. Intraocular anti-Leptospira antibody production was proven, leptospires could be cultured from the vitreous samples, and the LipL32 gene could be detected in the vitreous samples by PCR. Thus, there was convincing evidence of a chronic intraocular leptospiral infection, which can be eliminated most reliably by vitrectomy. Recently, it has been shown that the intraocular leptospires produce biofilm in the equine vitreous. Biofilm formation explains not only the success of vitrectomy, but also the survival of leptospires in the vitreous cavity for many years despite the presence of high intraocular antibody titers and immunocompetent cells, as well as the high tolerance to antibiotics.",book:{id:"11092",title:"Bacterial Biofilms",coverURL:"https://cdn.intechopen.com/books/images_new/11092.jpg"},signatures:"Bettina Wollanke and Hartmut Gerhards"},{id:"81758",title:"Growing Environmental Bacterium Biofilms in PEO Cryogels for Environmental Biotechnology Application",slug:"growing-environmental-bacterium-biofilms-in-peo-cryogels-for-environmental-biotechnology-application",totalDownloads:1,totalDimensionsCites:0,doi:"10.5772/intechopen.104813",abstract:"This Chapter discusses the entrapment, growing and biofilm formation by an environmental bacterium immobilized in polyethyleneoxide cryogel to be applied in environmental biotechnology. The KCM-R5 bacterium was isolated from the heavy metal-polluted environment near a large Pb-Zn smelter, also producing precious metals in Bulgaria. Molecular-genetic analysis revealed affiliation with Pseudomonas rhodesiae. The strain is capable of growing in high concentrations of phenol and different phenol derivatives. Polyethylene oxide was found to be friendly and nontoxic to bacteria polymer enabling bacteria easy to penetrate in it and fast to grow. KCM-R5 biofilms were grown for 30 days in batch culture with phenol (300-1000 mg L−1) dissolved in the mineral medium. The bacterium was able to involve phenol in its metabolism and use it as a single carbon supplier. The results obtained in the study showed 98% phenol biodegradation using the biotech installation described. The proposed PEO cryogel-P. rhodesiae KCM-R5 bacterium biotech biofilter can be used for environmental biotechnology application in industrial wastewater detoxification.",book:{id:"11092",title:"Bacterial Biofilms",coverURL:"https://cdn.intechopen.com/books/images_new/11092.jpg"},signatures:"Galina Satchanska"},{id:"81733",title:"Impairment of the Cardiovascular System during SARS-CoV-2 Infection",slug:"impairment-of-the-cardiovascular-system-during-sars-cov-2-infection",totalDownloads:3,totalDimensionsCites:0,doi:"10.5772/intechopen.103964",abstract:"Although the infection with the severe acute respiratory syndrome (SARS-CoV-2) virus affects primarily the respiratory system, it became evident from the very beginning that the coronavirus disease 2019 (COVID-19) is frequently associated with a large spectrum of cardiovascular involvements such as myocarditis/pericarditis, acute coronary syndrome, arrhythmias, or thromboembolic events, explained by a multitude of pathophysiological mechanisms. Individuals already suffering of significant cardiovascular diseases were more likely to be infected with the virus, had a worse evolution during COVID-19, with further deterioration of their basal condition and increased morbidity and mortality, but significant cardiac dysfunctions were diagnosed even in individuals without a history of heart diseases or being at low risk to develop such a pathology. Cardiovascular complications may occur anytime during the course of COVID-19, persisting even during recovery and, potentially, explaining many of the persisting symptoms included now in terms as subacute or long-COVID-19. It is now well accepted that in COVID-19, the occurrence of cardiovascular impairment represents a significant negative prognostic factor, immensely rising the burden of cardiovascular pathologies.",book:{id:"11369",title:"RNA Viruses",coverURL:"https://cdn.intechopen.com/books/images_new/11369.jpg"},signatures:"Cristina Tudoran, Mariana Tudoran, Voichita Elena Lazureanu, Adelina Raluca Marinescu, Dorin Novacescu and Talida Georgiana Cut"},{id:"81718",title:"Advances in the Development of Anti-Trichinella spiralis Vaccine, Challenges, and Future Prospective",slug:"advances-in-the-development-of-anti-trichinella-spiralis-vaccine-challenges-and-future-prospective",totalDownloads:6,totalDimensionsCites:0,doi:"10.5772/intechopen.103027",abstract:"Trichinellosis is a food-borne, zoonotic disease that causes infection by a nematode parasite belonging to the genus Trichinella. This is an important disease, and its causative agent is prevalent throughout the world (cosmopolitan). More clinical awareness of trichinellosis is required due to its many outbreaks, increase in the consumption of pork meat and its by-products. Trichinellosis is an epizootic in nature and its economic burden is associated with the prevention of this disease from the human food chain. This disease is transmitted from animals to humans through the consumption of raw or undercooked meat containing encapsulated muscle larvae of Trichinella spiralis. This paper demonstrates the direct effect of progesterone (P4) and mifepristone (RU486) on the progesterone receptors of T. spiralis. Also, studied the challenges in the preparation of DNA and recombinant protein vaccination to control trichinellosis. It is simply done this study at different life cycle developmental stages of T. spiralis. Vaccines development against T. spiralis infection is the new paradime shift from prevention of trichinellosis to fulfilling the food safety requirements.",book:{id:"11380",title:"Parasitic Helminths and Zoonoses - From Basic to Applied Research",coverURL:"https://cdn.intechopen.com/books/images_new/11380.jpg"},signatures:"Muhammad Tahir Aleem, Ruofeng Yan, Asad Khan, Rida Asrar, Amna Shakoor, Areej Asif, Zhaohai Wen, Zhengqing Yu, Muhammad Abdullah Malik, Tauseef-ur-Rehman, Rao Zahid Abbas, Muhammad Mohsin, Xiaokai Song, Lixin Xu and Xiangrui Li"},{id:"81699",title:"Efflux Pumps among Urinary E. coli and K. pneumoniae Local Isolates in Hilla City, Iraq",slug:"efflux-pumps-among-urinary-e-coli-and-k-pneumoniae-local-isolates-in-hilla-city-iraq",totalDownloads:4,totalDimensionsCites:0,doi:"10.5772/intechopen.104408",abstract:"Urinary tract infections (UTI) are the most common bacterial infections affecting humans. Escherichia coli and Klebsiella pneumoniae were common enterobacteria engaged with community-acquired UTIs. Efflux pumps were vital resistance mechanisms for antibiotics, especially among enterobacteria. Overexpression of an efflux system, which results in a decrease in antibiotic accumulation, is an effective mechanism for drug resistance. The ATP-binding cassette (ABC) transporters, small multidrug resistance (SMR), and multidrug and toxic compound extrusion (MATE) families, the major facilitator superfamily (MFS), and the resistance-nodulation- cell division (RND) family are the five superfamilies of efflux systems linked to drug resistance. This chapter highlights the results of studying the prevalence of efflux pump genes among local isolates of E. coli and K. pneumoniae in Hilla City, Iraq. class RND AcrAB-TolC, AcrAD-TolC, and AcrFE-TolC genes detected by conventional PCR of E. coli and K. pneumoniae respectively. The result revealed approximately all studied efflux transporter were found in both E. coli and K. pneumoniae in different percentages. Biofilm formation were observed in 50(100%) of K. pneumoniae and 49(98%) of E. coli isolates were biofilm former and follow: 30(60%), 20(40%) were weak, 12(24%), 22(44%) were moderate and 7(14%) and 8(16%) were Strong biofilm former for E. coli and K. pneumoniae, respectively.",book:{id:"11373",title:"The Global Antimicrobial Resistance Epidemic – Innovative Approaches and Cutting-Edge Solutions",coverURL:"https://cdn.intechopen.com/books/images_new/11373.jpg"},signatures:"Hussein Al-Dahmoshi, Sahar A. 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He holds a Master’s Degree in Urban Development Planning from the University College of London (UCL), and a Ph.D. in Urban Planning & Engineering from TU Berlin. He has conducted applied research on urban planning and infrastructure issues in over 20 countries in Africa and Asia. In 2005 he joined Eawag-Sandec as Leader of the Strategic Environmental Sanitation Planning Group. Since 2015 he heads the research department Sanitation, Water and Solid Waste for Development (Sandec) at the Swiss Federal Institute of Aquatic Research and Technology (Eawag).",institutionString:"Swiss Federal Institute of Aquatic Science and Technology, Switzerland",institution:null},editorTwo:{id:"290571",title:"Dr.",name:"Rui Alexandre",middleName:null,surname:"Castanho",slug:"rui-alexandre-castanho",fullName:"Rui Alexandre Castanho",profilePictureURL:"https://mts.intechopen.com/storage/users/290571/images/system/290571.jpg",biography:"Rui Alexandre Castanho has a master\\'s degree in Planning, Audit, and Control in Urban Green Spaces and an international Ph.D. in Sustainable Planning in Borderlands. Currently, he is a professor at WSB University, Poland, and a visiting professor at the University of Johannesburg, South Africa. Dr. Castanho is a post-doc researcher on the GREAT Project, University of Azores, Ponta Delgada, Portugal. He collaborates with the Environmental Resources Analysis Research Group (ARAM), University of Extremadura (UEx), Spain; VALORIZA - Research Center for the Enhancement of Endogenous Resources, Polytechnic Institute of Portalegre (IPP), Portugal; Centre for Tourism Research, Development and Innovation (CITUR), Madeira, Portugal; and AQUAGEO Research Group, University of Campinas (UNICAMP), Brazil.",institutionString:"University of Johannesburg, South Africa and WSB University, Poland",institution:{name:"University of Johannesburg",institutionURL:null,country:{name:"South Africa"}}},editorThree:null}]},overviewPageOFChapters:{paginationCount:2,paginationItems:[{id:"81644",title:"Perspective Chapter: Ethics of Using Placebo Controlled Trials for Covid-19 Vaccine Development in Vulnerable Populations",doi:"10.5772/intechopen.104776",signatures:"Lesley Burgess, Jurie Jordaan and Matthew Wilson",slug:"perspective-chapter-ethics-of-using-placebo-controlled-trials-for-covid-19-vaccine-development-in-vu",totalDownloads:4,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"SARS-CoV-2 Variants - Two Years After",coverURL:"https://cdn.intechopen.com/books/images_new/11573.jpg",subseries:{id:"6",title:"Viral Infectious Diseases"}}},{id:"80546",title:"Streptococcal Skin and Skin-Structure Infections",doi:"10.5772/intechopen.102894",signatures:"Alwyn Rapose",slug:"streptococcal-skin-and-skin-structure-infections",totalDownloads:48,totalCrossrefCites:0,totalDimensionsCites:0,authors:null,book:{title:"Streptococcal Infections",coverURL:"https://cdn.intechopen.com/books/images_new/10828.jpg",subseries:{id:"3",title:"Bacterial Infectious Diseases"}}}]},overviewPagePublishedBooks:{paginationCount:13,paginationItems:[{type:"book",id:"6667",title:"Influenza",subtitle:"Therapeutics and Challenges",coverURL:"https://cdn.intechopen.com/books/images_new/6667.jpg",slug:"influenza-therapeutics-and-challenges",publishedDate:"September 19th 2018",editedByType:"Edited by",bookSignature:"Shailendra K. 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Since then, he has been working as an Adjunct Professor in the same Department at the University of Pavia. His research activity during the first years was primarily focused on the purification and structural characterization of enzymes from animal and plant sources. During this period, Prof. Iadarola familiarized himself with the conventional techniques used in column chromatography, spectrophotometry, manual Edman degradation, and electrophoresis). Since 1995, he has been working on: i) the determination in biological fluids (serum, urine, bronchoalveolar lavage, sputum) of proteolytic activities involved in the degradation processes of connective tissue matrix, and ii) on the identification of biological markers of lung diseases. 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Biochemistry examines macromolecules - proteins, nucleic acids, carbohydrates, and lipids – and their building blocks, structures, functions, and interactions. Much of biochemistry is devoted to enzymes, proteins that catalyze chemical reactions, enzyme structures, mechanisms of action and their roles within cells. Biochemistry also studies small signaling molecules, coenzymes, inhibitors, vitamins, and hormones, which play roles in life processes. Biochemical experimentation, besides coopting classical chemistry methods, e.g., chromatography, adopted new techniques, e.g., X-ray diffraction, electron microscopy, NMR, radioisotopes, and developed sophisticated microbial genetic tools, e.g., auxotroph mutants and their revertants, fermentation, etc. More recently, biochemistry embraced the ‘big data’ omics systems. Initial biochemical studies have been exclusively analytic: dissecting, purifying, and examining individual components of a biological system; in the apt words of Efraim Racker (1913 –1991), “Don’t waste clean thinking on dirty enzymes.” Today, however, biochemistry is becoming more agglomerative and comprehensive, setting out to integrate and describe entirely particular biological systems. The ‘big data’ metabolomics can define the complement of small molecules, e.g., in a soil or biofilm sample; proteomics can distinguish all the comprising proteins, e.g., serum; metagenomics can identify all the genes in a complex environment, e.g., the bovine rumen. This Biochemistry Series will address the current research on biomolecules and the emerging trends with great promise.",coverUrl:"https://cdn.intechopen.com/series/covers/11.jpg",latestPublicationDate:"May 15th, 2022",hasOnlineFirst:!0,numberOfOpenTopics:4,numberOfPublishedChapters:286,numberOfPublishedBooks:27,editor:{id:"31610",title:"Dr.",name:"Miroslav",middleName:null,surname:"Blumenberg",fullName:"Miroslav Blumenberg",profilePictureURL:"https://mts.intechopen.com/storage/users/31610/images/system/31610.jpg",biography:"Miroslav Blumenberg, Ph.D., was born in Subotica and received his BSc in Belgrade, Yugoslavia. He completed his Ph.D. at MIT in Organic Chemistry; he followed up his Ph.D. with two postdoctoral study periods at Stanford University. Since 1983, he has been a faculty member of the RO Perelman Department of Dermatology, NYU School of Medicine, where he is codirector of a training grant in cutaneous biology. Dr. Blumenberg’s research is focused on the epidermis, expression of keratin genes, transcription profiling, keratinocyte differentiation, inflammatory diseases and cancers, and most recently the effects of the microbiome on the skin. He has published more than 100 peer-reviewed research articles and graduated numerous Ph.D. and postdoctoral students.",institutionString:null,institution:{name:"New York University Langone Medical Center",institutionURL:null,country:{name:"United States of America"}}},subseries:[{id:"14",title:"Cell and Molecular Biology",keywords:"Omics (Transcriptomics; Proteomics; Metabolomics), Molecular Biology, Cell Biology, Signal Transduction and Regulation, Cell Growth and Differentiation, Apoptosis, Necroptosis, Ferroptosis, Autophagy, Cell Cycle, Macromolecules and Complexes, Gene Expression",scope:"The Cell and Molecular Biology topic within the IntechOpen Biochemistry Series aims to rapidly publish contributions on all aspects of cell and molecular biology, including aspects related to biochemical and genetic research (not only in humans but all living beings). We encourage the submission of manuscripts that provide novel and mechanistic insights that report significant advances in the fields. Topics include, but are not limited to: Advanced techniques of cellular and molecular biology (Molecular methodologies, imaging techniques, and bioinformatics); Biological activities at the molecular level; Biological processes of cell functions, cell division, senescence, maintenance, and cell death; Biomolecules interactions; Cancer; Cell biology; Chemical biology; Computational biology; Cytochemistry; Developmental biology; Disease mechanisms and therapeutics; DNA, and RNA metabolism; Gene functions, genetics, and genomics; Genetics; Immunology; Medical microbiology; Molecular biology; Molecular genetics; Molecular processes of cell and organelle dynamics; Neuroscience; Protein biosynthesis, degradation, and functions; Regulation of molecular interactions in a cell; Signalling networks and system biology; Structural biology; Virology and microbiology.",annualVolume:11410,isOpenForSubmission:!0,coverUrl:"https://cdn.intechopen.com/series_topics/covers/14.jpg",editor:{id:"165627",title:"Dr.",name:"Rosa María",middleName:null,surname:"Martínez-Espinosa",fullName:"Rosa María Martínez-Espinosa",profilePictureURL:"https://mts.intechopen.com/storage/users/165627/images/system/165627.jpeg",institutionString:null,institution:{name:"University of Alicante",institutionURL:null,country:{name:"Spain"}}},editorTwo:null,editorThree:null,editorialBoard:[{id:"79367",title:"Dr.",name:"Ana Isabel",middleName:null,surname:"Flores",fullName:"Ana Isabel Flores",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRpIOQA0/Profile_Picture_1632418099564",institutionString:null,institution:{name:"Hospital Universitario 12 De Octubre",institutionURL:null,country:{name:"Spain"}}},{id:"328234",title:"Ph.D.",name:"Christian",middleName:null,surname:"Palavecino",fullName:"Christian Palavecino",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y000030DhEhQAK/Profile_Picture_1628835318625",institutionString:null,institution:{name:"Central University of Chile",institutionURL:null,country:{name:"Chile"}}},{id:"186585",title:"Dr.",name:"Francisco Javier",middleName:null,surname:"Martin-Romero",fullName:"Francisco Javier Martin-Romero",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bSB3HQAW/Profile_Picture_1631258137641",institutionString:null,institution:{name:"University of Extremadura",institutionURL:null,country:{name:"Spain"}}}]},{id:"15",title:"Chemical Biology",keywords:"Phenolic Compounds, Essential Oils, Modification of Biomolecules, Glycobiology, Combinatorial Chemistry, Therapeutic peptides, Enzyme Inhibitors",scope:"Chemical biology spans the fields of chemistry and biology involving the application of biological and chemical molecules and techniques. In recent years, the application of chemistry to biological molecules has gained significant interest in medicinal and pharmacological studies. This topic will be devoted to understanding the interplay between biomolecules and chemical compounds, their structure and function, and their potential applications in related fields. Being a part of the biochemistry discipline, the ideas and concepts that have emerged from Chemical Biology have affected other related areas. 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Behind these definitions are hidden all the aspects of normal and pathological functioning of all processes that the topic ‘Metabolism’ will cover within the Biochemistry Series. 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Thus proteomics, an area of research that detects all protein forms expressed in an organism, including splice isoforms and post-translational modifications, is more suitable than genomics for a comprehensive understanding of the biochemical processes that govern life. The most common proteomics applications are currently in the clinical field for the identification, in a variety of biological matrices, of biomarkers for diagnosis and therapeutic intervention of disorders. From the comparison of proteomic profiles of control and disease or different physiological states, which may emerge, changes in protein expression can provide new insights into the roles played by some proteins in human pathologies. Understanding how proteins function and interact with each other is another goal of proteomics that makes this approach even more intriguing. Specialized technology and expertise are required to assess the proteome of any biological sample. Currently, proteomics relies mainly on mass spectrometry (MS) combined with electrophoretic (1 or 2-DE-MS) and/or chromatographic techniques (LC-MS/MS). MS is an excellent tool that has gained popularity in proteomics because of its ability to gather a complex body of information such as cataloging protein expression, identifying protein modification sites, and defining protein interactions. The Proteomics topic aims to attract contributions on all aspects of MS-based proteomics that, by pushing the boundaries of MS capabilities, may address biological problems that have not been resolved yet.",annualVolume:11414,isOpenForSubmission:!0,coverUrl:"https://cdn.intechopen.com/series_topics/covers/18.jpg",editor:{id:"200689",title:"Prof.",name:"Paolo",middleName:null,surname:"Iadarola",fullName:"Paolo Iadarola",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bSCl8QAG/Profile_Picture_1623568118342",institutionString:null,institution:{name:"University of Pavia",institutionURL:null,country:{name:"Italy"}}},editorTwo:{id:"201414",title:"Dr.",name:"Simona",middleName:null,surname:"Viglio",fullName:"Simona Viglio",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRKDHQA4/Profile_Picture_1630402531487",institutionString:null,institution:{name:"University of Pavia",institutionURL:null,country:{name:"Italy"}}},editorThree:null,editorialBoard:[{id:"72288",title:"Dr.",name:"Arli Aditya",middleName:null,surname:"Parikesit",fullName:"Arli Aditya Parikesit",profilePictureURL:"https://mts.intechopen.com/storage/users/72288/images/system/72288.jpg",institutionString:null,institution:{name:"Indonesia International Institute for Life Sciences",institutionURL:null,country:{name:"Indonesia"}}},{id:"40928",title:"Dr.",name:"Cesar",middleName:null,surname:"Lopez-Camarillo",fullName:"Cesar Lopez-Camarillo",profilePictureURL:"https://mts.intechopen.com/storage/users/40928/images/3884_n.png",institutionString:null,institution:{name:"Universidad Autónoma de la Ciudad de México",institutionURL:null,country:{name:"Mexico"}}},{id:"81926",title:"Dr.",name:"Shymaa",middleName:null,surname:"Enany",fullName:"Shymaa Enany",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRqB9QAK/Profile_Picture_1626163237970",institutionString:null,institution:{name:"Suez Canal University",institutionURL:null,country:{name:"Egypt"}}}]}]}},libraryRecommendation:{success:null,errors:{},institutions:[]},route:{name:"profile.detail",path:"/profiles/76161",hash:"",query:{},params:{id:"76161"},fullPath:"/profiles/76161",meta:{},from:{name:null,path:"/",hash:"",query:{},params:{},fullPath:"/",meta:{}}}},function(){var e;(e=document.currentScript||document.scripts[document.scripts.length-1]).parentNode.removeChild(e)}()