Main advantages and inconveniences for canine AI.
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Barely three months into the new year and we are happy to announce a monumental milestone reached - 150 million downloads.
\n\nThis achievement solidifies IntechOpen’s place as a pioneer in Open Access publishing and the home to some of the most relevant scientific research available through Open Access.
\n\nWe are so proud to have worked with so many bright minds throughout the years who have helped us spread knowledge through the power of Open Access and we look forward to continuing to support some of the greatest thinkers of our day.
\n\nThank you for making IntechOpen your place of learning, sharing, and discovery, and here’s to 150 million more!
\n\n\n\n\n'}],latestNews:[{slug:"webinar-introduction-to-open-science-wednesday-18-may-1-pm-cest-20220518",title:"Webinar: Introduction to Open Science | Wednesday 18 May, 1 PM CEST"},{slug:"step-in-the-right-direction-intechopen-launches-a-portfolio-of-open-science-journals-20220414",title:"Step in the Right Direction: IntechOpen Launches a Portfolio of Open Science Journals"},{slug:"let-s-meet-at-london-book-fair-5-7-april-2022-olympia-london-20220321",title:"Let’s meet at London Book Fair, 5-7 April 2022, Olympia London"},{slug:"50-books-published-as-part-of-intechopen-and-knowledge-unlatched-ku-collaboration-20220316",title:"50 Books published as part of IntechOpen and Knowledge Unlatched (KU) Collaboration"},{slug:"intechopen-joins-the-united-nations-sustainable-development-goals-publishers-compact-20221702",title:"IntechOpen joins the United Nations Sustainable Development Goals Publishers Compact"},{slug:"intechopen-signs-exclusive-representation-agreement-with-lsr-libros-servicios-y-representaciones-s-a-de-c-v-20211123",title:"IntechOpen Signs Exclusive Representation Agreement with LSR Libros Servicios y Representaciones S.A. de C.V"},{slug:"intechopen-expands-partnership-with-research4life-20211110",title:"IntechOpen Expands Partnership with Research4Life"},{slug:"introducing-intechopen-book-series-a-new-publishing-format-for-oa-books-20210915",title:"Introducing IntechOpen Book Series - A New Publishing Format for OA Books"}]},book:{item:{type:"book",id:"8142",leadTitle:null,fullTitle:"Number Theory and Its Applications",title:"Number Theory and Its Applications",subtitle:null,reviewType:"peer-reviewed",abstract:"Number theory and its applications are well known for their proven properties and excellent applicability in interdisciplinary fields of science. Until now, research on number theory and its applications has been done in mathematics, applied mathematics, and the sciences. In particular, number theory plays a fundamental and important role in mathematics and applied mathematics. This book is based on recent results in all areas related to number theory and its applications.",isbn:"978-1-83968-051-9",printIsbn:"978-1-83968-050-2",pdfIsbn:"978-1-83968-052-6",doi:"10.5772/intechopen.78090",price:119,priceEur:129,priceUsd:155,slug:"number-theory-and-its-applications",numberOfPages:218,isOpenForSubmission:!1,isInWos:null,isInBkci:!1,hash:"90d1376ab2f3b8554ef8002ddf380da0",bookSignature:"Cheon Seoung Ryoo",publishedDate:"November 4th 2020",coverURL:"https://cdn.intechopen.com/books/images_new/8142.jpg",numberOfDownloads:5537,numberOfWosCitations:3,numberOfCrossrefCitations:2,numberOfCrossrefCitationsByBook:0,numberOfDimensionsCitations:2,numberOfDimensionsCitationsByBook:0,hasAltmetrics:0,numberOfTotalCitations:7,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"November 11th 2019",dateEndSecondStepPublish:"March 18th 2020",dateEndThirdStepPublish:"May 17th 2020",dateEndFourthStepPublish:"August 5th 2020",dateEndFifthStepPublish:"October 4th 2020",currentStepOfPublishingProcess:5,indexedIn:"1,2,3,4,5,6,7",editedByType:"Edited by",kuFlag:!1,featuredMarkup:null,editors:[{id:"230100",title:"Prof.",name:"Cheon Seoung",middleName:null,surname:"Ryoo",slug:"cheon-seoung-ryoo",fullName:"Cheon Seoung Ryoo",profilePictureURL:"https://mts.intechopen.com/storage/users/230100/images/system/230100.jpeg",biography:"Dr. Cheon Seoung Ryoo is a professor in mathematics at Hannam University. He received his Ph.D. in mathematics from Kyushu University. \n\nDr. Ryoo is the author of several research articles in numerical computations with guaranteed accuracy. Also, he has contributed to the field of scientific computing, p-adic functional analysis, and analytic number theory. More recently, he has been working with quantum calculus, special functions, differential equations, and dynamical systems.",institutionString:"Hannam University",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"3",totalChapterViews:"0",totalEditedBooks:"2",institution:{name:"Hannam University",institutionURL:null,country:{name:"Korea, South"}}}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"1399",title:"Discrete Mathematics",slug:"mathematics-discrete-mathematics"}],chapters:[{id:"73212",title:"A New Integer-to-Integer Transform",doi:"10.5772/intechopen.93356",slug:"a-new-integer-to-integer-transform",totalDownloads:434,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"This chapter presents a detailed analysis of an integer-to-integer transform that is closely related to the discrete Fourier transform, but that offers insights into signal structure that the DFT does not. The transform is analyzed for its underlying properties using concepts from number theory. Theorems are given along with proofs to help establish the salient features of the transform. Two kinds of redundancy exist in the transform. It is shown how redundancy implicit in the transform can be eliminated to obtain a simple form. Closed-form formulas for the forward and inverse transforms are presented.",signatures:"Rajesh Cherian Roy",downloadPdfUrl:"/chapter/pdf-download/73212",previewPdfUrl:"/chapter/pdf-preview/73212",authors:[null],corrections:null},{id:"72695",title:"Digit Sums and Infinite Products",doi:"10.5772/intechopen.92365",slug:"digit-sums-and-infinite-products",totalDownloads:388,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"Consider the sequence un defined as follows: un=+1 if the sum of the base b digits of n is even, and un=−1 otherwise, where we take b=2. Recall that the Woods-Robbins infinite product involves a rational function in n and the sequence un. Although several generalizations of the Woods-Robbins product are known in the literature, no other infinite product involving a rational function in n and the sequence un was known in closed form until recently. In this chapter we introduce a systematic approach to these products, which may be generalized to other values of b. We illustrate the approach by evaluating a large class of similar infinite products.",signatures:"Samin Riasat",downloadPdfUrl:"/chapter/pdf-download/72695",previewPdfUrl:"/chapter/pdf-preview/72695",authors:[null],corrections:null},{id:"72767",title:"The Borel-Cantelli Lemmas, and Their Relationship to Limit Superior and Limit Inferior of Sets (or, Can a Monkey Really Type Hamlet?)",doi:"10.5772/intechopen.93121",slug:"the-borel-cantelli-lemmas-and-their-relationship-to-limit-superior-and-limit-inferior-of-sets-or-can",totalDownloads:575,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"The purpose of this chapter is to show that if a monkey types infinitely, Shakespeare’s Hamlet and any other works one may wish to add to the list will each be typed, not once, not twice, but infinitely often with a probability of 1. This dramatic fact is a simple consequence of the Borel-Cantelli lemma and will come as no surprise to anyone who has taken a graduate-level course in Probability. The proof of this result, however, is quite accessible to anyone who has but a rudimentary understanding of the concept of independence, together with the notion of limit superior and limit inferior of a sequence of sets.",signatures:"Anant P. Godbole",downloadPdfUrl:"/chapter/pdf-download/72767",previewPdfUrl:"/chapter/pdf-preview/72767",authors:[null],corrections:null},{id:"73384",title:"Prime Numbers Distribution Line",doi:"10.5772/intechopen.92639",slug:"prime-numbers-distribution-line",totalDownloads:445,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"During the analysis of the fractal-primorial periodicity of the natural series of numbers, presented in the form of an alternation (sequence) of prime numbers (1 smallest prime factor > 1 of any integer), the regularity of prime numbers distribution was revealed. That is, the theorem is proved that for any integer = N on the segment of the natural series of numbers from 1 to N + 2N: (1) prime numbers are arranged in groups, by exactly three consecutive prime numbers of the form: (Р1-Р2-Р3). In this case, the distance from the first to the third prime number of any group is less than 2N integers, that is, Р3–Р1 < 2N integers. (2) These same prime numbers are redistributed in a line in groups, by exactly two consecutive prime numbers, on all segments of the natural series of numbers shorter than 2Nintegers.",signatures:"Shcherbakov Aleksandr Gennadiyevich",downloadPdfUrl:"/chapter/pdf-download/73384",previewPdfUrl:"/chapter/pdf-preview/73384",authors:[null],corrections:null},{id:"71899",title:"Moments of Catalan Triangle Numbers",doi:"10.5772/intechopen.92046",slug:"moments-of-catalan-triangle-numbers",totalDownloads:542,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"In this chapter, we consider the Catalan numbers, \n\n\nC\nn\n\n=\n\n1\n\nn\n+\n1\n\n\n\n\n\n\n2\nn\n\n\n\n\nn\n\n\n\n\n\n, and two of their generalizations, Catalan triangle numbers, \n\n\nB\n\nn\n,\nk\n\n\n\n and \n\n\nA\n\nn\n,\nk\n\n\n\n, for \n\nn\n,\nk\n∈\nN\n\n. They are combinatorial numbers and present interesting properties as recursive formulae, generating functions and combinatorial interpretations. We treat the moments of these Catalan triangle numbers, i.e., with the following sums: \n\n\n∑\n\nk\n=\n1\n\nn\n\n\nk\nm\n\n\nB\n\nn\n,\nk\n\nj\n\n,\n\n∑\n\nk\n=\n1\n\n\nn\n+\n1\n\n\n\n\n\n2\nk\n−\n1\n\n\nm\n\n\nA\n\nn\n,\nk\n\nj\n\n,\n\n for \n\nj\n,\nn\n∈\nN\n\n and \n\nm\n∈\nN\n∪\n\n0\n\n\n. We present their closed expressions for some values of \n\nm\n\n and \n\nj\n\n. Alternating sums are also considered for particular powers. Other famous integer sequences are studied in Section 3, and its connection with Catalan triangle numbers are given in Section 4. Finally we conjecture some properties of divisibility of moments and alternating sums of powers in the last section.",signatures:"Pedro J. Miana and Natalia Romero",downloadPdfUrl:"/chapter/pdf-download/71899",previewPdfUrl:"/chapter/pdf-preview/71899",authors:[null],corrections:null},{id:"72952",title:"Modular Sumset Labelling of Graphs",doi:"10.5772/intechopen.92701",slug:"modular-sumset-labelling-of-graphs",totalDownloads:378,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"Graph labelling is an assignment of labels or weights to the vertices and/or edges of a graph. For a ground set X of integers, a sumset labelling of a graph is an injective map f:VG→PX such that the induced function f⊕:EG→PX is defined by f+uv=fu+fv, for all uv∈EG, where fu+fv is the sumset of the set-label, the vertices u and v. In this chapter, we discuss a special type of sumset labelling of a graph, called modular sumset labelling and its variations. We also discuss some interesting characteristics and structural properties of the graphs which admit these new types of graph labellings.",signatures:"Sudev Naduvath",downloadPdfUrl:"/chapter/pdf-download/72952",previewPdfUrl:"/chapter/pdf-preview/72952",authors:[null],corrections:null},{id:"72114",title:"Determination of the Properties of (p, q)‐Sigmoid Polynomials and the Structure of Their Roots",doi:"10.5772/intechopen.91862",slug:"determination-of-the-properties-of-p-q-sigmoid-polynomials-and-the-structure-of-their-roots",totalDownloads:395,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"Nowadays, many mathematicians have great concern about \n\n\np\nq\n\n\n-numbers, which are various applications, and have studied these numbers in many different research areas. We know that \n\n\np\nq\n\n\n-numbers are different to \n\nq\n\n-numbers because of the symmetric property. We find the addition theorem, recurrence formula, and \n\n\np\nq\n\n\n-derivative about sigmoid polynomials including \n\n\np\nq\n\n\n-numbers. Also, we derive the relevant symmetric relations between \n\n\np\nq\n\n\n-sigmoid polynomials and \n\n\np\nq\n\n\n-Euler polynomials. Moreover, we observe the structures of appreciative roots and fixed points about \n\n\np\nq\n\n\n-sigmoid polynomials. By using the fixed points of \n\n\np\nq\n\n\n-sigmoid polynomials and Newton’s algorithm, we show self-similarity and conjectures about \n\n\np\nq\n\n\n-sigmoid polynomials.",signatures:"Jung Yoog Kang",downloadPdfUrl:"/chapter/pdf-download/72114",previewPdfUrl:"/chapter/pdf-preview/72114",authors:[null],corrections:null},{id:"71745",title:"I–Convergence of Arithmetical Functions",doi:"10.5772/intechopen.91932",slug:"i-convergence-of-arithmetical-functions",totalDownloads:601,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"Let \n\nn\n>\n1\n\n be an integer with its canonical representation, \n\nn\n=\n\np\n1\n\nα\n1\n\n\n\np\n2\n\nα\n2\n\n\n⋯\n\np\nk\n\nα\nk\n\n\n\n. Put \n\nH\n\nn\n\n=\nmax\n\n\nα\n1\n\n…\n\nα\nk\n\n\n\n, \n\nh\n\nn\n\n=\nmin\n\n\nα\n1\n\n…\n\nα\nk\n\n\n\n, \n\nω\n\nn\n\n=\nk\n\n, \n\nΩ\n\nn\n\n=\n\nα\n1\n\n+\n⋯\n+\n\nα\nk\n\n\n, \n\nf\n\nn\n\n=\n\n∏\n\nd\n∣\nn\n\n\nd\n\n and \n\n\nf\n∗\n\n\nn\n\n=\n\n\nf\n\nn\n\n\nn\n\n\n. Many authors deal with the statistical convergence of these arithmetical functions. For instance, the notion of normal order is defined by means of statistical convergence. The statistical convergence is equivalent with \n\n\nI\nd\n\n\n–convergence, where \n\n\nI\nd\n\n\n is the ideal of all subsets of positive integers having the asymptotic density zero. In this part, we will study \n\nI\n\n–convergence of the well-known arithmetical functions, where \n\nI\n=\n\nI\nc\n\nq\n\n\n=\n\n\nA\n⊂\nN\n:\n\n∑\n\na\n∈\nA\n\n\n\na\n\n−\nq\n\n\n<\n+\n∞\n\n\n\n is an admissible ideal on \n\nN\n\n such that for \n\nq\n∈\n\n0\n1\n\n\n we have \n\n\nI\nc\n\nq\n\n\n⊊\n\nI\nd\n\n\n, thus \n\n\nI\nc\n\nq\n\n\n\n–convergence is stronger than the statistical convergence (\n\n\nI\nd\n\n\n–convergence).",signatures:"Vladimír Baláž and Tomáš Visnyai",downloadPdfUrl:"/chapter/pdf-download/71745",previewPdfUrl:"/chapter/pdf-preview/71745",authors:[null],corrections:null},{id:"72027",title:"Identification of Eigen-Frequencies and Mode-Shapes of Beams with Continuous Distribution of Mass and Elasticity and for Various Conditions at Supports",doi:"10.5772/intechopen.92185",slug:"identification-of-eigen-frequencies-and-mode-shapes-of-beams-with-continuous-distribution-of-mass-an",totalDownloads:906,totalCrossrefCites:1,totalDimensionsCites:1,hasAltmetrics:0,abstract:"In the present article, an equivalent three degrees of freedom (DoF) system of two different cases of inverted pendulums is presented for each separated case. The first case of inverted pendulum refers to an amphi-hinge pendulum that possesses distributed mass and stiffness along its height, while the second case of inverted pendulum refers to an inverted pendulum with distributed mass and stiffness along its height. These vertical pendulums have infinity number of degree of freedoms. Based on the free vibration of the above-mentioned pendulums according to partial differential equation, a mathematically equivalent three-degree of freedom system is given for each case, where its equivalent mass matrix is analytically formulated with reference on specific mass locations along the pendulum height. Using the three DoF model, the first three fundamental frequencies of the real pendulum can be identified with very good accuracy. Furthermore, taking account the 3 × 3 mass matrix, it is possible to estimate the possible pendulum damages using a known technique of identification mode-shapes via records of response accelerations. Moreover, the way of instrumentation with a local network by three accelerometers is given via the above-mentioned three degrees of freedom.",signatures:"Triantafyllos K. Makarios",downloadPdfUrl:"/chapter/pdf-download/72027",previewPdfUrl:"/chapter/pdf-preview/72027",authors:[{id:"69418",title:"Prof.",name:"Triantafyllos",surname:"Makarios",slug:"triantafyllos-makarios",fullName:"Triantafyllos Makarios"}],corrections:null},{id:"72360",title:"Some Identities Involving 2-Variable Modified Degenerate Hermite Polynomials Arising from Differential Equations and Distribution of Their Zeros",doi:"10.5772/intechopen.92687",slug:"some-identities-involving-2-variable-modified-degenerate-hermite-polynomials-arising-from-differenti",totalDownloads:554,totalCrossrefCites:0,totalDimensionsCites:0,hasAltmetrics:0,abstract:"In this chapter, we introduce the 2-variable modified degenerate Hermite polynomials and obtain some new symmetric identities for 2-variable modified degenerate Hermite polynomials. In order to give explicit identities for 2-variable modified degenerate Hermite polynomials, differential equations arising from the generating functions of 2-variable modified degenerate Hermite polynomials are studied. Finally, we investigate the structure and symmetry of the zeros of the 2-variable modified degenerate Hermite equations.",signatures:"Cheon Seoung Ryoo",downloadPdfUrl:"/chapter/pdf-download/72360",previewPdfUrl:"/chapter/pdf-preview/72360",authors:[{id:"230100",title:"Prof.",name:"Cheon Seoung",surname:"Ryoo",slug:"cheon-seoung-ryoo",fullName:"Cheon Seoung Ryoo"}],corrections:null},{id:"73172",title:"Elliptic Curve over a Local Finite Ring Rn",doi:"10.5772/intechopen.93476",slug:"elliptic-curve-over-a-local-finite-ring-em-r-sub-n-sub-em-",totalDownloads:319,totalCrossrefCites:1,totalDimensionsCites:1,hasAltmetrics:0,abstract:"The goal of this chapter is to study some arithmetic proprieties of an elliptic curve defined by a Weierstrass equation on the local ring Rn=FqX/Xn, where n≥1 is an integer. It consists of, an introduction, four sections, and a conclusion. In the first section, we review some fundamental arithmetic proprieties of finite local rings Rn, which will be used in the remainder of the chapter. The second section is devoted to a study the above mentioned elliptic curve on these finite local rings for arbitrary characteristics. A restriction to some specific characteristic cases will then be considered in the third section. Using these studies, we give in the fourth section some cryptography applications, and we give in the conclusion some current research perspectives concerning the use of this kind of curves in cryptography. We can see in the conclusion of research in perspectives on these types of curves.",signatures:"Abdelhakim Chillali and Lhoussain El Fadil",downloadPdfUrl:"/chapter/pdf-download/73172",previewPdfUrl:"/chapter/pdf-preview/73172",authors:[null],corrections:null}],productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},subseries:null,tags:null},relatedBooks:[{type:"book",id:"8599",title:"Polynomials",subtitle:"Theory and Application",isOpenForSubmission:!1,hash:"1c36cace7f309250a777c0f404a2d79f",slug:"polynomials-theory-and-application",bookSignature:"Cheon Seoung Ryoo",coverURL:"https://cdn.intechopen.com/books/images_new/8599.jpg",editedByType:"Edited by",editors:[{id:"230100",title:"Prof.",name:"Cheon Seoung",surname:"Ryoo",slug:"cheon-seoung-ryoo",fullName:"Cheon Seoung Ryoo"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"1591",title:"Infrared Spectroscopy",subtitle:"Materials Science, Engineering and Technology",isOpenForSubmission:!1,hash:"99b4b7b71a8caeb693ed762b40b017f4",slug:"infrared-spectroscopy-materials-science-engineering-and-technology",bookSignature:"Theophile Theophanides",coverURL:"https://cdn.intechopen.com/books/images_new/1591.jpg",editedByType:"Edited by",editors:[{id:"37194",title:"Dr.",name:"Theophile",surname:"Theophanides",slug:"theophile-theophanides",fullName:"Theophile Theophanides"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3161",title:"Frontiers in Guided Wave Optics and Optoelectronics",subtitle:null,isOpenForSubmission:!1,hash:"deb44e9c99f82bbce1083abea743146c",slug:"frontiers-in-guided-wave-optics-and-optoelectronics",bookSignature:"Bishnu Pal",coverURL:"https://cdn.intechopen.com/books/images_new/3161.jpg",editedByType:"Edited by",editors:[{id:"4782",title:"Prof.",name:"Bishnu",surname:"Pal",slug:"bishnu-pal",fullName:"Bishnu Pal"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"3092",title:"Anopheles mosquitoes",subtitle:"New insights into malaria vectors",isOpenForSubmission:!1,hash:"c9e622485316d5e296288bf24d2b0d64",slug:"anopheles-mosquitoes-new-insights-into-malaria-vectors",bookSignature:"Sylvie Manguin",coverURL:"https://cdn.intechopen.com/books/images_new/3092.jpg",editedByType:"Edited by",editors:[{id:"50017",title:"Prof.",name:"Sylvie",surname:"Manguin",slug:"sylvie-manguin",fullName:"Sylvie Manguin"}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null,productType:{id:"1",chapterContentType:"chapter",authoredCaption:"Edited by"}},{type:"book",id:"371",title:"Abiotic Stress in Plants",subtitle:"Mechanisms and Adaptations",isOpenForSubmission:!1,hash:"588466f487e307619849d72389178a74",slug:"abiotic-stress-in-plants-mechanisms-and-adaptations",bookSignature:"Arun Shanker and B. 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Her research interests include renewable sources of energy from microalgae and further utilizing the residual biomass for the generation of value-added products, bioremediation through microalgae and microbial consortium, antioxidative enzymes and stress, and nutraceuticals from microalgae. She has been working on algal biotechnology for the last two decades. She has published her research in many international journals and has authored many books and chapters with renowned publishing houses. She has also delivered talks as an invited speaker at many national and international conferences. 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Artificial Insemination, one of the earliest techniques for assisted reproduction in animals and humans, took longer to be implemented in dogs due to species-specific particularities. In past decades, progresses in the knowledge of canine physiology and new advances in canine semen technology allowed these services to become available worldwide. Hence, subsequent to the increase in the artificial insemination demand among dog breeders and owners and the broaden of the AI to preserved semen as a management tool in canine breeding, as through international exchange of frozen semen, inbreeding within breeds can be reduced. Therefore, with spread of canine AI dog, breeders now may select stud dogs from all over the world to improve their kennel´ genetics, without transport-associated stress to the animals. Also, it is possible to save semen from valuable dogs into sperm bank to be used in next generations, after their death or the peak of reproductive age. In addition, breeders also are aware of the sanitary benefits associated with AI. Avoiding direct contact between the male and female, AI also prevents the spread of sexually transmitted diseases, as those originated by
Although the first reports on AI in dogs subsequent to the Spallanzani experiments (in late XVIII century) appeared by the end of the fifties, reporting the use of fresh semen, or in the sixties, the use of frozen semen, only in the nineties this technique was introduced into dog breeding practice, particularly in USA and Nordic countries (Foote, 2002; England & Millar, 2008). The reproductive physiology of this species and unfavourable response of the dog sperm to freezing were the two major constraints to the initial efforts to improve the AI technique in dogs (Linde Forsberg, 2005a). A lot of research was performed in those areas, especially in the northern Europe, to overcome these issues, generating a large amount of information and allowing technical development, in particular in the canine semen technology. Nowadays, as a consequence of the demand for reproductive technologies, in particular the AI with fresh or refrigerate semen, this is a current service offered in the small animal veterinary practice.
\n\t\t\tAccording to Linde-Forsberg (2001, 2005a), from all the AI in dogs performed by veterinarians today in Europe, about 50-55% is done with fresh semen, collected at the clinic, 10% with chilled semen and around 35-40% with frozen semen. However, at least in Portugal, the use of imported chilled semen is far most frequent than the use of frozen semen when compared to other countries in Northern Europe.
\n\t\t\tResearch on AI in the domestic dog, along with other reproductive technologies, proceed worldwide, particularly on sperm survival at freezing and the identification of deleterious components to spermatozoa or fertilization, providing important information for the preservation of wild
Several main indications exist to perform AI in the dog (Linde Forsberg, 2005a; England & Millar, 2008; Farstad, 2010). In parallel, some ethical conditions must be discussed when facing the different interests of specific groups, namely dogs, breeders, owners and veterinarians.
\n\t\t\tMain indications for AI in dogs include both medical and breeding-management reasons (Table 1). As major potential advantage, AI may allow to reduce physical distances, the use of genetically valuable stud dog semen all over the word, fighting the stress of transportation of animals and inbreeding (Johnston et al., 2001; Linde-Forsberg, 2005a). It is also an important technique whenever physical and behavioural abnormalities in the male or female preventing natural mating (Table 2).
\n\t\tPerforming canine AI may raise some ethical concerns, mostly to central institutions like the National Kennel Clubs or Veterinarian Orders or equivalent, in particular on what concerns the use of frozen semen and the need for intra-uterine insemination, mainly those involving surgical procedures. In fact, several countries (such as Norway, Sweden and the United Kingdom) refer to welfare concerns and discourage or even forbid the use of surgical procedures to obtain intra-uterine insemination (England & Millar, 2008; Linde-Forsberg, 2005b).
\n\t\t\tEthical issues are seldom associated with the non-surgical process of artificial insemination
Restrictions to the use of AI in animals that never matted despite all physiological conditions met together to guarantee a successful mate, may respond to the ethical issue that demands for ruling out clinical reasons for AI, as an underlying unaware problem (congenital or behavioural) may exists. This concern is in fact previewed in the Fédération Cynologique International (FCI) breeding rules (
Potential benefits | \n\t\t\t\t\t\tPotential weakness | \n\t\t\t\t\t
Decrease stress, infectious deseases transmission, travel expenses Semen collection without interruption of the male activity (show or training) Splitting of an ejaculate to bred more females Reduction of the costs with maintenance of stud dogs in a colony Worldwide availability of the semen of a given dog Allow early castration of working dogs while maintaining availability of their genes Evaluation of semen quality prior to AI Early detection of male reproductive pathologies Semen preservation, so genetic material may be available in the future Overcome problems associated with the refusal to breed (psychological or physical reasons, precocious ejaculation), inexperienced males May overcome quarantine restrictions | \n\t\t\t\t\t\tInduction of physical or psychological trauma during the AI process Risk of performing AI for inappropriate reasons Failure in careful clinical examination of the breeding animals Potential risk for maintaining some disorders in a particular genetic line (hip dysplasia or anatomical abnormality of the reproductive tract) Potential risk for introduction of inherited diseases or abnormalities Potential overuse of a given male within a programme or breed May allow confusion of parentage | \n\t\t\t\t\t
Main advantages and inconveniences for canine AI.
Factor | \n\t\t\t\t\t\tCause | \n\t\t\t\t\t
Female-dependent | \n\t\t\t\t\t\tMale rejection Aggression Congenital abnormalities (e.g. presence of a vaginal septum; genital tract strictures, small vulva and vagina) | \n\t\t\t\t\t
Male-dependent | \n\t\t\t\t\t\tReduced libido (due to local or systemic diseases, age, deficient breeding management, drugs) Pain Physical deficiency (such as inability to mount or to obtain penile erection, lumbar muscle problems, congenital abnormalities) | \n\t\t\t\t\t
Associated to both male and female | \n\t\t\t\t\t\tInexperience Male to female disproportion Social and behavioural problems (dominant female, inversion of the social hierarchy) | \n\t\t\t\t\t
Main causes for refusal of natural mating.
The competence of the operator to perform the procedures is essential to avoid all technique-related ethical constraints to the use of AI in dogs. Before offering canine AI services, practitioners ought to specialised themselves, acquiring profound knowledge of the reproductive physiology and pathology of the species and the skills to collect semen and to inseminate the female without risking animal health or welfare.
\n\t\tSemen collection in the dog is a relatively easy procedure, although requiring some training for optimization of the technique. Semen collection and evaluation is necessary to obtain good results in canine AI. Although practitioners are often asked to collect semen and perform AI without detailed semen analysis, every sample of semen collected should be evaluated (at least progressive forward motility, total sperm count and morphology) before it is used for artificial insemination or cryopreservation. Semen evaluation prior to insemination warrants the male potential fertility and consequently may predict the fertility potential for the AI. In addition, when preparing semen preservation, fertility certificate may be needed. In such cases andrological evaluation of the stud dog (breeding soundness evaluation or BSE) has to be performed. Semen collection should be performed before the physical exam or any stressful procedures on the stud, or can be booked to another day (Freshman, 2002; Johnston et al, 2001).
\n\t\t\t\tSemen can be collected from most dogs in the absence of a teaser, in a quiet and isolated room, where interruptions should be prevented, although the presence of a bitch would allow better ejaculates. In reluctant males, stimulating estrus scent can be provided by the presence of a female in estrus or by using frozen-thawed swabs or gauze sponges taken from vaginal secretions of estrus bitches (Freshman, 2002; Kutzler, 2005; Olson & Husted, 1986). Although possible, not everyone achieves the use of a chemical pheromone (methyl p-hydroxybenzoate, Aldrich Chemical, Milwaukee, WI) swabbed on the perineal area and tail of an anestrus teaser (Johnston et al., 2001; Kutzler, 2005).
\n\t\t\t\tCollection of semen should be prepared in advance, and interval between collections or between the natural mating and collection, should be registered, if the male is regularly used. Ideal intervals between collections are 2 to 5 days, whilst intervals longer than 10 days may result in an increased number of morphological abnormalities and decreased motility (Freshman, 2002; Johnston et al., 2001). In longer periods, it is advisable to perform one previous collection, if semen is to be chilled or frozen for shipment. If semen preservation is planned, semen extender should be prepared before the arrival of the animal (Freshman, 2002).
\n\t\t\t\tThe most common method for semen collection in the dog is by digital manipulation, in the presence of a female. However, bitch presence, although desirable as it facilitates procedures, is not essential to accomplish the collection (Farstad, 2010; Linde Forsberg, 2005a). It should be noticed that when the collection is achieved in the presence of the bitch ejaculates present higher concentration.
\n\t\t\t\tThe use of manual massage is the most commonly used technique (Farstad, 2010; Johnston et al., 2001; Linde Forsberg, 2005a), although in the past semen was collected from dogs using an artificial vagina. Nowadays, semen collection into a tube is commonly accomplished by penile massage and the use of a cone or plastic sleeve, a funnel or a special collecting vial (Linde Forsberg, 2005a). Briefly, the process is started with a massage of the dog prepuce at the level of the
Canine ejaculate consists of 3 fractions, with the first and third fraction consisting of prostatic fluid and the second being rich in spermatozoa (England et al., 1990) (Table 3). The first fraction, the presperm portion, is emitted in 0.5 to 1 minute and is colourless, with a volume range of 1-5 ml. It is expelled during first stage of erection, at the moment of the presence of evident copulatory movement of male. The second fraction, the sperm-rich portion, is also rapidly completed (1-2 minutes), and is grayish-white in colour, with a volume of 1-3 ml. It is expelled when thrusting movement of the male ceases and full erection is observed. The third fraction comes from the prostate and may be up to 30-40 ml; it may take up from 5 to 30 minutes to be completed (Günzel-Apel, 1994; Johnston et al., 2001).
\n\t\t\t\tCharacteristics | \n\t\t\t\t\t\t\t1st fraction | \n\t\t\t\t\t\t\t2nd Fraction | \n\t\t\t\t\t\t\t3rd Fraction | \n\t\t\t\t\t\t
Volume | \n\t\t\t\t\t\t\t0.1-2 mL (average 0.33 mL) | \n\t\t\t\t\t\t\t0.1-3 mL (average 1.17 mL) Sometimes larger volume | \n\t\t\t\t\t\t\t1-2 to "/20 mL Quite variable depending on the animal. | \n\t\t\t\t\t\t
Colour | \n\t\t\t\t\t\t\tclear or opaque | \n\t\t\t\t\t\t\tgreyish-white, white, milky-white | \n\t\t\t\t\t\t\tclear, transparent | \n\t\t\t\t\t\t
Consistency | \n\t\t\t\t\t\t\twatery | \n\t\t\t\t\t\t\twatery-milky, milky | \n\t\t\t\t\t\t\twatery | \n\t\t\t\t\t\t
Character | \n\t\t\t\t\t\t\tprostate secretion with admixture of epithelial cells, urine, bacteria and sperm cells | \n\t\t\t\t\t\t\tsperm cells suspended in seminal plasma | \n\t\t\t\t\t\t\tprostate gland secretion | \n\t\t\t\t\t\t
pH (average) | \n\t\t\t\t\t\t\t6.37 | \n\t\t\t\t\t\t\t6.10 | \n\t\t\t\t\t\t\t7.20 | \n\t\t\t\t\t\t
Duration | \n\t\t\t\t\t\t\t5-90 sec. (average 13.5 sec) | \n\t\t\t\t\t\t\t5-300 sec. (average 52.4 sec.) | \n\t\t\t\t\t\t\t60 sec-20 min. (average 6 min. 55 sec.) | \n\t\t\t\t\t\t
Main characteristics of the different fractions of the dog ejaculate.
Size of the dog | \n\t\t\t\t\t\t\tVolume of the ejaculate | \n\t\t\t\t\t\t
< 20 kg | \n\t\t\t\t\t\t\t1-22.5 mL (average 5.38 mL) | \n\t\t\t\t\t\t
"/ 20 kg | \n\t\t\t\t\t\t\t2-45 mL (average 12.75 mL) | \n\t\t\t\t\t\t
Variation on the volume of the ejaculate with the size of the dog (Dubiel, 2004)
In the dog, the volume of whole ejaculate varies between breeds (Table 4) mainly with animal size and is partially dependent on the volume of the third fraction collected, which constitute about 95% of the volume of the ejaculate in dogs (Farstad, 2010).
\n\t\t\t\tIn most dogs, semen can be collected twice at 30 minutes interval (Farstad, 2010), although the second sample is usually slightly diluted.
\n\t\t\t\tMost often, artificial insemination with freshly collected semen is performed without fractioning the ejaculate, although for artificial insemination, only the second fraction is of interest (Thomassen & Farstadt 2009; Root Kustritz, 2003). Furthermore, it has been demonstrated the existence of detrimental effects on fertility when this fraction is not separated from the second one, particularly if semen will be processed as chilled or frozen. Consequently, ejaculate fractioning should always be accomplished, particularly separation of the third fraction. If the ejaculate has a very small volume, it may be diluted with semen extender, to facilitate its handling during insemination procedures.
\n\t\t\tSemen assessment is an important part of the evaluation of fertility in males and it should be performed as routine element of prebreeding examination. Furthermore, semen evaluation ought to be completed before artificial insemination or sperm preservation. Semen should be assessed immediately after collection and it has to be handled carefully during all the procedures. Rapid changes of environmental temperature may be deleterious for spermatozoal motility and structure, and may also artifactually influence the results of examination. Any delay in semen assessment may decrease the percentage of motile sperm and simultaneously increase the percentage of dead sperm. It is advisable to keep all equipment necessary for semen collection and evaluation at the temperature near 37°C (Christiansen, 1984; Feldman & Nelson, 1996; Linde-Forsberg, 1991).
\n\t\t\t\tOn table 5, the most frequent indications for routine semen evaluation are presented. Semen evaluation is also frequently performed in the absence of known reproductive pathology, upon request of the owner. In addition, it can be performed at a predetermined moment after the diagnosis of a clinical disease that may have negative reflects on the potential fertility of a male dog.
\n\t\t\t\tIt should be notice that reliable
Most frequent indications | \n\t\t\t\t\t\t\tOther situations, on request | \n\t\t\t\t\t\t
Semen evaluation before artificial insemination Semen evaluation before/after chilling or cryopreservation Clinical signs of a disorder of male genital organs Whenever infertility or subfertility of a male is suspected | \n\t\t\t\t\t\t\tNew stud dog introduced to the breeding colony Evaluation of young stud dog before first mating In cases of serial unsuccessful matings of particular dog Pathological lesions of male genital tract observed by the owner Re-evaluation after the treatment of diseases of male genital tract | \n\t\t\t\t\t\t
Common reasons for canine semen assessment
The semen assessment performed once is not always reliable, because:
\n\t\t\t\tFrequent matings or semen collections may temporary result in a decreased semen quality;
After a prolonged sexual rest dogs may ejaculate many dead, immotile spermatozoa of abnormal morphology;
In young inexperienced males and dogs which mated earlier only naturally, without experience on semen collection, the obtained semen sample may contain only the part of sperm-rich fraction.
Different approaches are available to assess the quality of the dog semen that can be grouped in conventional and advanced techniques. The later, usually requires more sophisticated means for the semen assessment and the support of a technical equipment, while the former may be performed in an inhouse lab.
\n\t\t\t\t\tThe conventional approaches to semen evaluation include macroscopical evaluation of the semen (volume and colour), but also the microscopical assessment, which will give the concentration and the number of viable cells in the ejaculate.
\n\t\t\t\t\tVolume. The volume of the ejaculate may be assessed in the calibrated tubes used for semen collection. It mainly dependends on the size of the dog, the size of the prostate gland, the animal age, the frequency of semen collection, the level of erotisation, and the volume of 3rd fraction collected. A decrease of semen volume is observed in cases of benign prostatic hyperplasia, prostatic cysts, inflammatory lesions of prostate and testicles, inflammation of epididymis, vas deferens or urethra and at weak libido.
\n\t\t\t\t\t\tColour. The colour of whole ejaculate depends on the volume of third fraction of ejaculate collected, on the concentration of spermatozoa per mL and the potential presence of non-germ cells in the ejaculate. When analysing the colour, one should be aware of the method of collection, as colour varies with the fraction to be analysed and the fact that analysis may been performed on the whole semen or on fractioned semen. The normal colour of whole ejaculate is greyish-white. Pathological colours include: green-greyish typical for the presence of the pus in semen; red or pink-specific for erythrocytes contamination (haemorrhages from urethra or corpora cavernosa, prostatitis); yellow specific for urine contamination; and brown, if in the presence of blood.
\n\t\t\t\t\t\tAny kind of semen contamination, such as hair or mud, exclude the specimen from further procedures including artificial insemination or semen preservation. It is therefore important to check the region of praeputial opening before semen collection and to clean it.
\n\t\t\t\t\t\tThe presence of sediment consisting of sperm cells at the bottom of the tube is a normal feature if the semen is left for several minutes.
\n\t\t\t\t\tMotility. One of the most important step of conventional semen assessment is the subjective evaluation of progressively motile spermatozoa (Spz) under contrast-phase microscope. The optimal temperature for assessment of dog sperm cell motility is 39°C. A small drop of about 20 µL of semen is placed on in a pre-warmed slide and cover by the coverslip. The evaluation is performed under the objective of x20 to x40. If the highly concentrated sperm-rich fraction is collected separately, the semen should be extended with saline or Tris-buffer to a concentration allowing the observation of particular, single sperm cells. The assessment is based on the evaluation of the average percentage of progressively motile spermatozoa in a few different fields of the specimen. The normal dog semen contains at least 70% of progressively motile spermatozoa (Feldman & Nelson, 1996; Günzel-Apel, 1994).
\n\t\t\t\t\t\tA decrease in the percentage of motile spermatozoa may results from temperature shock, contamination with water, urine, blood or lubricants but also from long sexual abstinence and systemic or infectious diseases, such as brucellosis. Sperm agglutination is always pathological and is frequently found in cases of infectious diseases.
\n\t\t\t\t\t\tConcentration and total sperm count. The sperm concentration in whole canine normal ejaculate usually exceeds 80 x106 Spz/mL. If the second fraction of ejaculate is collected separately, the sperm cells concentration in sperm-rich fraction varies usually between 200-600 x 106 Spz/mL. It is generally assumed that the number of motile spermatozoa necessary for successful AI should be >150 x106 (Linde-Forsberg, 1991). Therefore, under normal conditions, the dog´s ejaculate contains far more sperm cells than those needed for a seminal dose, although sometimes, especially in miniature or toy breeds, ejaculate volume and the total number of sperm cells are relatively low (<100 x106 Spz/mL). The concentration of spermatozoa in semen volume is usually assessed by cytometric method on the haemocytometer, such as the Thoma, Thoma-Neu, Bürker or Neubauer chambers, with semen pre-diluted at 1:200. In order to find the sperm count
A large variety in the total number of spermatozoa
Sperm morphology. The percentage of morphologically normal spermatozoa in canine semen should be greater than 70% (Günzel-Apel, 1994). The morphology may be assessed under contrast-phase microscope, but usually the evaluation is performed under light microscope on stained slides. Smears of undiluted or diluted ejaculate are examined microscopically for the presence of structural abnormalities of spermatozoa. The stains used include modified Giemsa stain (DiffQuik) and Spermac® stain. The semen is smeared on a glass slide in a similar manner to that of blood, air dried and stained. The semen may be also stained with a nigrosin-eosin stain. A drop of this stain is gently mixed with a drop of semen on a pre-warmed slide before being smeared, and allowed to air dry. Evaluation of sperm morphology should be completed microscopically using oil immersion, using an objective of x100 or x 125. A minimum number of 200 spermatozoa should be counted and evaluated for the presence of abnormalities. The percentage of cells with particular morphological defects and of normal cells are calculated. Traditionally sperm cells abnormalities are divided into primary defects - originating from abnormalities of spermatogenesis and secondary defects - originating from abnormalities of semen maturation, transit through the ductal system and specimen preparation. According to another classification sperm abnormalities may be divided into major defects, negatively correlated with fertility, and minor defects, unassociated with fertility (Table 6) (Oettle, 1993).
\n\t\t\t\t\t\t\n\t\t\t\t\t\t\t\t\t | Primary spermatozoa defects | \n\t\t\t\t\t\t\t\t\tSecondary spermatozoa defects | \n\t\t\t\t\t\t\t\t
Head | \n\t\t\t\t\t\t\t\t\tMacrocephalus, microcephalus, double, pointed, indented heads | \n\t\t\t\t\t\t\t\t\tFree, bent heads, swollen acrosomes, detaching acrosomes | \n\t\t\t\t\t\t\t\t
Neck | \n\t\t\t\t\t\t\t\t\tThickened, eccentric insertion | \n\t\t\t\t\t\t\t\t\t\n\t\t\t\t\t\t\t\t |
Midpiece | \n\t\t\t\t\t\t\t\t\tThickened, thinned, coiled, kinked, double midpiece | \n\t\t\t\t\t\t\t\t\tBent midpiece, extraneous material surrounding midpiece, proximal, mid and distal cytoplasmatic droplets | \n\t\t\t\t\t\t\t\t
Tail | \n\t\t\t\t\t\t\t\t\tThin, double, triple tail. | \n\t\t\t\t\t\t\t\t\tCoiled, looped, kinked, folded, detached tail. | \n\t\t\t\t\t\t\t\t
Main defects of the dog spermatozoa
The acrosomal status, which is frequently assessed for the estimation of the quality of the frozen-thawed semen, may be evaluated with the use of eosin-nigrosin, Giemsa, Trypan blue, Bismarck brown, Rose Bengal or Spermac® stainings (Dahlbom et al.,1997; Dott & Foster, 1972; Watson, 1975). When a spermatozoon presents more than one abnormality, it should be classified according to the most important abnormality or with the most prevalent one, if they have equal significance (Oettle, 1993).
\n\t\t\t\t\t\t‘Live-dead’ spermatozoa. The assessment of the percentage of live and dead spermatozoa is based on the assumption that dead spermatozoa possess disintegrated plasma membrane allowing eosin penetration. Thus the percentage of eosin positive cells stained with nigrosin-eosin stain is considered as percentage of dead cells. The normal dog semen consists of maximal percentage of 30% of dead sperm cells. The evaluation of the percentage of live and dead spermatozoa and the percentage of morphological defects may be performed on the same nigrosin-eosin stained slides.
\n\t\t\t\t\tIn the past 2 to 3 decades, several strategies were developed to escape the subjectivity in the semen evaluation, related to the experience and skills of the observer, the method of specimen preparation, staining technique and number of cells evaluated, and wich is particularly important when the fertility potential of preserved sperm cells has to be ascertain. It is well documented that variations in results of the conventional evaluation of the same semen samples obtained by different observers and laboratories may reach 30-60% (Coetzee et al., 1999; Davis & Katz, 1992). Moreover, implementation of such methodologies, not routinely usable in the small to median veterinary clinics due to their costs, allows accurate comparisons between laboratories worlwide and minimizes occurence of large errors. Furthermore, advanced semen assessment is essential whenever the semen has to be preserved, in particular for freezing. Advanced semen assessment techniques are sumarized on table 7. In general, the results obtained with these methods are better correlated with the AI outcome than the results of traditional semen evaluation.
\n\t\t\t\t\tTests | \n\t\t\t\t\t\t\t\tAims | \n\t\t\t\t\t\t\t\tProcedure and Analysis | \n\t\t\t\t\t\t\t\tReferences | \n\t\t\t\t\t\t\t
Hypo-osmotic swelling test. | \n\t\t\t\t\t\t\t\tSpz membrane integrity (indirect method) | \n\t\t\t\t\t\t\t\tSperm incubation with hypo-osmotic solutions for 30 minutes at 37°C Spz with intact plasmalemma become swolled and show coiled tails | \n\t\t\t\t\t\t\t\tEngland & Plummer, 1993; Kumi-Diaka, 1993 | \n\t\t\t\t\t\t\t
Computer assisted sperm analysis [CASA] | \n\t\t\t\t\t\t\t\tObjective evaluation of sperm cell motility | \n\t\t\t\t\t\t\t\tDetermination of motility parameters for individual spz Characterization of Spz movement according to the average velocity, the trajectory, the amplitude of movement and beat cross frequency. It allows identification of Spz subpopulations | \n\t\t\t\t\t\t\t\tVerstegen et al., 2001; Rijsselaere et al., 2003; Niżański et al., 2009 | \n\t\t\t\t\t\t\t
Zona pellucida binding assay | \n\t\t\t\t\t\t\t\tAssessment of sperm fertilizing potential | \n\t\t\t\t\t\t\t\t- ZP-binding assay ( ZBA) using intact homologous oocytes - hemizona binding assay (HZA) using bisected hemizonae The number of spermatozoa bound to ZP is counted with contrast-phase microscopy. The number of bound Spz reflects its fertilizing potential | \n\t\t\t\t\t\t\t\tHermansson et al., 2006; Kawakami et al., 1998; Rijsselaere et al., 2005; Ström-Holst et al., 2001 | \n\t\t\t\t\t\t\t
Fluorescent probes and flow cytometry | \n\t\t\t\t\t\t\t\tMembrane integrity and evaluation of live and dead cells | \n\t\t\t\t\t\t\t\tCombined use of several fluorescent dyes (i.e, propidium iodide PI and carboxyfluorescein diacetate, SYBR-14/PI) allow the identification of live cells Live cells activate fluorescence (deacylation) which is maintained intracellularly in intact membrane cells. Dead Spz are stained red due to the influx of PI through damaged plasma membrane. | \n\t\t\t\t\t\t\t\tHewitt & England, 1998; Peña et al., 1998; Rijsselaere et al, 2005; P.F. Silva & Gadella, 2006 | \n\t\t\t\t\t\t\t
Capacitation status | \n\t\t\t\t\t\t\t\tFluorescent antibiotic chlorotetracycline (CTC), when bound to free calcium ions, is fluorescent. Combined with Hoechst 33258 allows also assessment of percentage of live cells and capacitation status Three classes of sperm cells may be assessed: uncapacitated and acrosome intact (F-pattern), capacitated and acrosome intact (B-pattern) and capacitated and acrosome reacted (AR-pattern) | \n\t\t\t\t\t\t\t\tGuérin et al., 1999; Hewitt & England, 1998; Petrunkina et al., 2004; Rota et al. 1999b | \n\t\t\t\t\t\t\t|
Acrosomal status | \n\t\t\t\t\t\t\t\tLectins conjugated with fluorescein isothiocyanate, such as Peanut Agglutinin (FITC-PNA) or Pisum Sativum Agglutinin (FITC-PSA). PNA labelling is specific for the outer acrosomal membrane whereas PSA is labelling acrosomal matrix. The absence of the fluorescence of the living sperm indicates an intact acrosome, whereas the presence of the fluorescence is indicative for acrosome disruption or acrosome reaction | \n\t\t\t\t\t\t\t\tKawakami et al., 2002; Peña et al., 2001; Sirivaidyapong et al., 2000; P.F. Silva & Gadella, 2006 | \n\t\t\t\t\t\t\t|
Mitochondria | \n\t\t\t\t\t\t\t\tRhodamine 123 (R123) is a potentiometric membrane dye used for the selective staining of functional mitochondria. It fluoresces only when the proton gradient over the inner mitochondrial membrane (IMM) is built up | \n\t\t\t\t\t\t\t\tGarner et al., 1997; Gravance et al., 2001 | \n\t\t\t\t\t\t\t|
Integrity of DNA structure | \n\t\t\t\t\t\t\t\t- Sperm chromatin structure assay (SCSA) with acridine orange (AO). - Terminal deoxynucleotidyl transferase-mediated nick end labeling (TUNEL) The SCSA is a flow cytometric method for identification of changes in the DNA status. AO shows green fluorescence when DNA is intact and red fluorescence when DNA is denaturated | \n\t\t\t\t\t\t\t\tChohan et al., 2006; Bochenek et al., 2001; Garcia-Macis et al, 2006 | \n\t\t\t\t\t\t\t
Concise description of the available advanced methods for sperm quality assessment.
The key-issues to obtain good results by using canine artificial insemination are:
\n\t\t\tProper timing of the insemination
The use of adequate number of viable sperm cells
Good semen preparation and handling
Adequate deposition of semen in the female reproductive tract
On next sections the major issues on timing the AI and available techniques of semen deposition on the bitch genital tract will be discussed.
\n\t\t\tObtaining successful pregnancies and adequate number of offspring per litter depends upon the correct timing for mating, as well as for insemination, particularly because bitches are mono-estrous, presenting usually one to two reproductive cycles per year. Although relationship among behavioral, hormonal and physiological events for the average bitch exists, considerable individual variation are also currently found on what concerns the duration of the estrogenic and early luteal stages (proestrus and estrus) and of the anestrus (Concannon et al., 1977; Concannon, 2004). The bitch usually presents a relatively long follicular phase and considerable variability exists in the onset of estrous behavior and acceptance of the male, making it difficult to determine occurrence of the LH surge and onset of ovulation in this species unless specific methods for timing the ovulation and estimating the fertile period are used (Linde-Forsberg, 1991). Furthermore, in this species, ovulation of immature oocytes (primary oocytes, before extrusion of the first polar body) determines the need for a maturation period in distal oviducts that may last for 96-108 hours (Concannon, 2004, 2010; Tsuitsui, 1989; Tsutsui et al., 2009); for most bitches, the secondary oocytes present a life span of 24-48h (Tsuitsui, 1989). Those particularities in the reproductive physiology may explain why the major cause for infertility in the bitch is the inappropriate breeding management (Goodman, 2001; Linde-Forsberg, 1991). Consequently, careful planning of mating time by timing ovulation is a key step in canine artificial insemination.
\n\t\t\tDetermination of blood progesterone and the vaginal cell cornification on cytological specimens are the most widely used techniques (Linde Forsberg, 2003), to which recently has been added the vaginal endoscopy (that replaces the vaginoscopic exam) and the ultrasonographic follow-up of the follicular development and ovulation (England & Concannon, 2002; Hewitt & England, 2000; Fontbonne & Malandain, 2006; Levy & Fontbonne, 2007). These evaluations should be performed in sequence and with 2-3 days intervals for the majority of females (if the bitch has been reported to present short heat period, of about 6 to 9 days, is possible that daily evaluations may be needed).
\n\t\t\t\tOn the vaginal cytology, epithelial cells of the vagina change their form in response to estrogen impregnation, and passes from small round cells with a clearly visible cytoplasm in non-estrogenic stages, to larger, cornified, angular shaped-cells with small pyknotic nucleus, almost to the point of disappearing, under the influence of estrogens (Figure 1). At beginning of estrus, vaginal cytology presents its maximum cornification index (>70%). By that time, serial blood sampling for progesterone determination should start to detect the initial progesterone rise (2-3 ng/mL) which correlates with LH surge, which in turn triggers ovulation within 2 days. On the day of ovulation (day “0” of the cycle) progesterone concentrations may vary between 4 and 10 ng/mL. The sudden increase in the number of round-shaped cells and of neutrophils reflects the onset of diestrus (Fontbonne & Malandain, 2006).
\n\t\t\t\tProgesterone semi-quantitative immunoenzymatic assays are available for clinical routines, but although rapids, these test lack accuracy. They give progesterone concentration according to a colorimetric scale for values corresponding to basal progesterone levels (0-1 ng/mL), intermediate levels corresponding to the LH surge (around 1-2.5ng/mL) and the ovulation periods (2.5 – 8ng/mL), and high progesterone levels (more than 8 or 10, depending on the kit). A recent study showed that, in dogs, semi-quantitative methods for progesterone determination are less accurate than the quantitative methods, in particular at intermediate plasma progesterone concentrations (Moxon et al., 2010). According to this study, the tested semi-quantitative assay estimated higher progesterone concentration than
\n\t\t\t\tRIA (radioimmunoassay), which could suggest that the fertilization period had commenced earlier than it was actually the case. In addition to those assays, quantitative radio or chemiluminescent assays can also be used, even if not always available in the house lab, since cross-reactivity exist to the molecule between different species, for example with human progesterone.
\n\t\t\t\tSchematic representation of the major morphological changes of the predominant epithelial cell in the vaginal cytology during the dog estrous cycle. On the bottom, images of the vaginal péri-estrus cytological preparations stained with Harris-Shorr.
Although ovarian ultrasound examination is a reliable and accurate method to determine ovulation in most domestic females, in the bitch fat accumulation in ovarian bursa that encloses the ovaries may difficult the value of the technique. In addition, several studies demonstrated that ultrasound images of the ovaries around ovulation are more difficult to analyze, due to the fact that ovarian follicles do not differ much in the immediate pre- and post-ovulatory period (England & Concannon, 2002), as not all dog follicles collapse at ovulation (Yeager & Concannon, 1996) and also because non-ovulated follicles frequently remain in the ovary (Wallace et al. 1992).
\n\t\t\t\t\tConsequently, follicular dynamics evaluation through ultrasonography (US) in dogs is still experimental and must follow a very precise protocol, which accuracy increase with the use of frequent examinations. In a recent study, Fontbonne (2008) reported that US was accurate enough to detect the occurrence of ovulation and obtain comparable numbers of ovarian structures between US examinations and macroscopic visual count on the surface of the ovaries after surgical removal, even if only one daily examination was performed. However, that author accepts that features of ovulation may be difficult to visualize in large breeds and in overweight animals. Pre-ovulatory follicles may present different aspects at US. Usually they appear as round to slightly triangular anechoic structures, sometimes slightly flattened, giving a honeycomb aspect to the ovary (Figure 2). At ovulation, different degrees of follicular collapse can be found in the US images, and usually a clear change of the ovarian echogenicity has been detected in a large number of bitches, giving the ovary a more homogeneous aspect (Fontbonne, 2008; Fontbonne & Malandain, 2006). Persistence of non-ovulatory follicular structures was perceived in US images after ovulation. Also, in the immediate post-ovulation period, until 24 hours after US changes of the ovaries at ovulation, hypoechoic structures were observed in most cases (Figure 2). These structures were very similar to the pre-ovulatory follicles, although slightly smaller, and tending to increase in echogenicity (from the border to the interior of the structure) with time (Fontbonne, 2008; Fontbonne & Malandain, 2006).
\n\t\t\t\t\tUltrasonographic scans of canine ovaries before and after LH surge and ovulation. US are compared with images of longitudinal sections of canine ovaries of similar stages of follicle developement.
It is possible to use vaginal endoscopy to determine the fertile period although it does not allow accurate timing of ovulation. However, this method requires expensive equipment. Nevertheless, it may give a huge contribution to the vaginal evaluation and detection of anatomical abnormalities that may impair proper reproductive performance.
\n\t\t\t\t\tThe fluctuation of estrogen and progesterone concentrations in the blood at consecutive stages of estrous cycle in the bitch results in specific morphologic changes of the vaginal mucosa. Analysis of these changes allows for exact assessment of the stage of the estrous cycle and for determination of the optimal insemination time (Goodman, 2001; Jeffcoate & Lindsay, 1989; Lindsay 1983). The observation of the cranial part of the vagina is performed for this purpose. The deep introduction of the tip of endoscope into the narrow part of the vagina close to the cervix (dorsal median postcervical fold) or paracervix, is of less diagnostic value (Pineda et al., 1973).
\n\t\t\t\t\tVaginoscopic examination is performed using a rigid endoscope 3-4 mm in diameter, with diagnostic sheath and a length of 30-33 cm or longer. The examination should be done on the standing animal. Usually there is no need of administration of sedatives. The tip of the endosope is introduced at the beginning at angle of 45-60° cranially and dorsally. When the tip of the optics reaches the vagina it should be repositioned at horizontal axis.
\n\t\t\t\t\tDuring proestrus the increase of the estrogen concentration results in the oedema of the vaginal mucosa. Vaginoscopy reveals rounded folds in the vagina. The mucosa of the folds is turgid, pink in colour and with a smooth surface. Normally the bloody discharge is also visible in the vagina. Sometimes, periodic blood outflow from the cervix, through the paracervix may be observed. The lumen of the vagina is narrow, which can be appreciated when the endoscope is advanced cranially. At the last days of proestrus and at beginning of estrus, the decrease of estrogen concentration and increase of progesterone (P4) level is noted. It results in the collapse of vaginal folds. Formerly turgid and smooth, the mucosa, becomes wrinkled and shrunked. Vaginal folds become smaller. Maximal intensity of shrinkage of vaginal mucosa is observed between 3 and 7-8 days of estrous cycle. This time the loss of fluid from the tissue of vaginal mucosa and submucosa is great and the shape of vaginal folds become angulated with sharp angles at the top of folds. As the result, the lumen of the vagina is wider in comparison to proestrus. During diestrus vaginal folds become flat and round. The mucosa is red and small petechia may be visible at places touched by the tip of the endoscope. This is due to the fact that epithelium of the vagina is thin and consists of only 2-3 cell layers in diestrus and anestrus. An opaque, thick mucus is sometimes visible on the surface of epithelium (Figure 3).
\n\t\t\t\tOther methods has been proposed to monitor the bitch oestrous cycle, such as serial reading of the electrical resistance of the vaginal mucus around the time of ovulation, using probes inserted into the vagina during the heat period (Fontbonne, 2008), or the crystallization patterns in anterior vaginal fluids (England & Allen, 1989) or in saliva (Pardo-Carmona et al., 2010), which have been found up not to present an acceptable reliabily in the identification of the canine ovulation.
\n\t\t\t\t\tVaginal endoscopy of the bitch. [From left to right] Aspect of the vaginal folds at early proestrus, proestrus, estrus and diestrus.
In the bitch, when timing the day of ovulation as accurately as possible is essential to guarantee adequate fertility in natural mating systems, it becomes even more important to determine precisely when to inseminate bitches according to the sort of semen to be used (fresh, chilled or frozen semen), as usually semen longevity and sperm cells survival decreases with time. In addition, in frozen/thawed semen sperm cell capacitation is shorter due to secondary effects of the frozen procedure.
\n\t\t\t\tWhen fresh or chilled semen is used, insemination should be performed on the day of ovulation, and a second insemination must be schedule for 2 days later. On the contrary, when frozen/thawed semen is used, and considering the need of canine oocytes to mature in the oviducts, insemination should be performed 2 days after ovulation, and the second insemination 48h later (Figure 4). However, scheduling for the artificial inseminations may be slightly adjusted according to the experience of the operator, the place for semen deposition and the limitation on the number of inseminations. Consequently, regimes for canine AI may vary with authors (Root Kustritz, 2003). Table 8 condenses the available information on the AI schedules for fresh, chilled and frozen semen.
\n\t\t\t\tGraphic representation of the fertile period and the ideal moment for canine AI according to the type of semen.
Semen | \n\t\t\t\t\t\t\tDosis | \n\t\t\t\t\t\t\tExpected spz survival | \n\t\t\t\t\t\t\tInsemination schedule | \n\t\t\t\t\t\t\tExpected fertility | \n\t\t\t\t\t\t
Fresh | \n\t\t\t\t\t\t\t150-200x106 spz/mL (extended) | \n\t\t\t\t\t\t\t4-6 days | \n\t\t\t\t\t\t\tEvery other day, when P4 rise above 4ng/mL, up to 3 times. Day 1 to 4 post-ovulation P4 levels between 8 and 15ng/mL | \n\t\t\t\t\t\t\t80-90%(either with transcervical or vaginal deposition) | \n\t\t\t\t\t\t
Chilled | \n\t\t\t\t\t\t\t150 - 200x106 spz/mL (extended) | \n\t\t\t\t\t\t\t24-72hrs | \n\t\t\t\t\t\t\tBreeding once or twice 2-4 days post ovulation (P4 = 4 -10ng/mL). Day 2 to 4 post-ovulation P4 levels between 8 and 15ng/mL | \n\t\t\t\t\t\t\t80-90% (either with transcervical or vaginal deposition) | \n\t\t\t\t\t\t
Frozen | \n\t\t\t\t\t\t\t50 - 300x106 spz/mL (extended) | \n\t\t\t\t\t\t\t12-24hrs. | \n\t\t\t\t\t\t\tTwice, at P4 levels above 8ng/mL and estrus vaginal cytology Day 5 to 7 post-ovulation P4 levels between 18 and 28 ng/mL | \n\t\t\t\t\t\t\t45% if vaginal deposition 67 -84% if transcervical or intrauterine | \n\t\t\t\t\t\t
Artificial insemination schedules for dogs, according to the type of semen used.
If in the AI with fresh semen the success of the procedure is strongly related to the quality of semen used and the moment for AI (Table 9), when using chilled semen both the quality of semen and the site of semen deposition are important factors for success, whilst in the AI with frozen semen, the intra-uterine semen deposition is critical (Table 10).
\n\t\t\t\t\n\t\t\t\t\t\t\t | 2AI, 48h apart | \n\t\t\t\t\t\t\tMultiple AI | \n\t\t\t\t\t\t\t2AI, day 3 and 5 post LH surge | \n\t\t\t\t\t\t|||||||
Day 9-13 after bloody vaginal discharge | \n\t\t\t\t\t\t\tEosinophilic index "/80% | \n\t\t\t\t\t\t\tEosinophilic index "/80% | \n\t\t\t\t\t\t\tProgesterone measurements | \n\t\t\t\t\t\t|||||||
n | \n\t\t\t\t\t\t\t% | \n\t\t\t\t\t\t\tn | \n\t\t\t\t\t\t\t% | \n\t\t\t\t\t\t\tn | \n\t\t\t\t\t\t\t% | \n\t\t\t\t\t\t\tn | \n\t\t\t\t\t\t\t% | \n\t\t\t\t\t\t|||
Pregnancy rates | \n\t\t\t\t\t\t\t29 | \n\t\t\t\t\t\t\t55.8a | \n\t\t\t\t\t\t\t33 | \n\t\t\t\t\t\t\t66.0 ab | \n\t\t\t\t\t\t\t37 | \n\t\t\t\t\t\t\t78.7 b | \n\t\t\t\t\t\t\t37 | \n\t\t\t\t\t\t\t80.4 b | \n\t\t\t\t\t\t||
Whelping rates | \n\t\t\t\t\t\t\t27 | \n\t\t\t\t\t\t\t51.9a | \n\t\t\t\t\t\t\t31 | \n\t\t\t\t\t\t\t62.0 a | \n\t\t\t\t\t\t\t36 | \n\t\t\t\t\t\t\t76.6 b | \n\t\t\t\t\t\t\t36 | \n\t\t\t\t\t\t\t78.3 b | \n\t\t\t\t\t\t||
Length of pregnancy | \n\t\t\t\t\t\t\t63.7±1.4a | \n\t\t\t\t\t\t\t64.5±2.6b | \n\t\t\t\t\t\t\t64.4±2.4b | \n\t\t\t\t\t\t\t63.1±1.3c | \n\t\t\t\t\t\t||||||
Number per group | \n\t\t\t\t\t\t\t52 | \n\t\t\t\t\t\t\t50 | \n\t\t\t\t\t\t\t47 | \n\t\t\t\t\t\t\t46 | \n\t\t\t\t\t\t
a,b,c – different superscript mean significant differences (P<0.05) between groups
Success of canine AI with fresh semen according to the method used to timing AI (n=136) (Adapted from Niżański et al. 2005)
Semen | \n\t\t\t\t\t\t\tSperm deposition | \n\t\t\t\t\t\t|
Intra-vaginal (n) | \n\t\t\t\t\t\t\tIntra-uterine (n) | \n\t\t\t\t\t\t|
Fresh | \n\t\t\t\t\t\t\t47.7 (1212) | \n\t\t\t\t\t\t\t62 (121) | \n\t\t\t\t\t\t
Chilled | \n\t\t\t\t\t\t\t45.4 (348) | \n\t\t\t\t\t\t\t65.0 (40) | \n\t\t\t\t\t\t
Frozen | \n\t\t\t\t\t\t\t36.7 (30) | \n\t\t\t\t\t\t\t55.5 (290) | \n\t\t\t\t\t\t
Comparison of whelping rates (%) in vaginal and intrauterine inseminations according to the type of dog semen (n=2041) (Adapted from Linde Forsberg, 2002a).
In dogs, during natural mating, occurs the projection of a considerable portion of the ejaculate into the uterus, through the cervical canal, during the coital tie (England et al., 2006; Thomassen & Farstad, 2009). When performing AI we should be aware that vaginal deposition
As in other species, in dogs sperm cell number in the uterine lumen may be influenced by many factors, such as the moment of estrus, the type of breeding (natural mating or insemination) the method of insemination (intravaginal or intrauterine), the type of semen (fresh, chilled or frozen) and sperm quality (total and progressive motility and sperm speed), besides some individual variations (Rijsselaere et al., 2004). However, despite the influence of the intrauterine
Based on the last two years inseminations performed at the clinic (with a global success rate of around 75%), using a comercial kit for progesterone determination, fresh semen and intra-vaginal deposition (2 AI, 48h apart), it was found that when inseminations were performed with progesterone levels above 8 ng/mL a higher success rate and a closer to predicted whelping dates were achieved (Table 11), comparing to inseminations at lower levels of progesterone (2.5-8 ng/mL). No diferences were found in the litter size between these groups, which were very similar in age and parity of the bitches.
\n\t\t\t\tSemen | \n\t\t\t\t\t\t\tNumber of animals | \n\t\t\t\t\t\t\tRange of age (mean) | \n\t\t\t\t\t\t\tProgesterone at AI (ng/ml) | \n\t\t\t\t\t\t\tDays from last IA to whelping | \n\t\t\t\t\t\t\tSuccess rate(n° females whelped /inseminated) | \n\t\t\t\t\t\t\tLiter size(± SD) | \n\t\t\t\t\t\t|
Fresh | \n\t\t\t\t\t\t\t39 | \n\t\t\t\t\t\t\t18 | \n\t\t\t\t\t\t\t1-6 years (2.94 years) | \n\t\t\t\t\t\t\t2.5 to 8 ng/mL | \n\t\t\t\t\t\t\t63 | \n\t\t\t\t\t\t\t66.6 | \n\t\t\t\t\t\t\t6.33± 2.84 | \n\t\t\t\t\t\t
12 | \n\t\t\t\t\t\t\t1-6 years (3.23 years) | \n\t\t\t\t\t\t\t"/ 8 ng/mL | \n\t\t\t\t\t\t\t61 | \n\t\t\t\t\t\t\t80.95 | \n\t\t\t\t\t\t\t6.29 ± 2.26 | \n\t\t\t\t\t\t||
Chilled | \n\t\t\t\t\t\t\t42 | \n\t\t\t\t\t\t\t1-6 years (3.6 years) | \n\t\t\t\t\t\t\t"/8 ng/mL | \n\t\t\t\t\t\t\t61 | \n\t\t\t\t\t\t\t73.8 | \n\t\t\t\t\t\t\t6.1 ±2.51 | \n\t\t\t\t\t\t
Results for the AI procedures with fresh and chilled semen and 2 AI
Independently of the place for semen deposition, repeating the AI at 24-48 hrs intervals results in a significantly higher fertility: for fresh semen both the pregnancy rate and litter size present a significantly increase when multiple AI are performed (Linde-Forsberg & Forsberg, 1993), whilst for frozen semen the differences on the pregnancy rates are not significant, although litter size tended to increase with the number of inseminations (Linde-Forsberg, 2000, 2002a).
\n\t\t\t\tDeep vaginal insemination is probably the widestly used method for insemination with fresh semen when the technique is performed by the breeder or in small budget clinics. For vaginal AI a simple plastic catheter of proper length may be used, to which a plastic disposable syringe containing the semen is attached. Or a commercial catheter in flexible latex tube presenting an inflatable balloon at the tip, like the Osiris gun, may be used; when inflated, this kind of device has the advantage of increasing the probability for intrauterine transport of the semen and of preventing semen backflow (Farstadt, 2010; Linde Forsberg, 2005a).
\n\t\t\t\t\tBefore AI procedures start, cleaning of the perineal area, in particular the peri-vulvar area, is needed. As transabdominal palpation is usually used to guide or ascertain the vaginal catheter position, the owner of the female should be instructed to bring the animal with an empty stomach, which facilitates the procedure (Linde Forsberg, 2005a).
\n\t\t\t\t\tThe bitch is placed in a standing position on an examination table or on the floor (according to the size of the female). To avoid catheterization of the urethra (the urethral opening in the bitch is located at the pelvic brim), particular attention should be paid not to unintentionally introduce into the urinary bladder. The insemination catheter is carefully introduced in the vagina of the bitch, first steeply upwards until the pelvic brim has been passed, and then in a horizontal angle, when it is carefully pushed further ahead (Farstad, 2010). In alternative, the vulva may be elevated to just below the anus (as the bitch does when stimulated by the male dog) (Linde Forsberg, 2005a). At this point, the position of the AI catheter must be learn by palpation, and orientated. If the catheter is in the urinary bladder, the cranial part of the vagina and the cervix may be palpable above the catheter and also the tip of the catheter stands out more clearly, due to the thinner walls of the urinary bladder in comparison to those of the vagina (Linde Forsberg, 2005a). After certification that the catheter is correctly placed, it is moved onward through the cranial portion of the vagina delimited by the dorsal medial folds. In smaller or primiparous bitches this point can be difficult to overcome, and may not be possible to pass the catheter into the cervical fornix. Except for those females, the AI catheter should be further introduced until it reaches the paracervical area, which can be palpated as a 1- to 2-cm-long, firm structure that ends at the cervix (a firm, rounded to ovoid structure, freely movable). The semen is deposited once the catheter has been located in the paracervical area, close to the external
During AI the bitch is held with the hindquarters up and head down, in an angle of 45-60°. This position facilitates transabdominal palpation of the cervix and ensures that the semen will not be expelled through backflow. According to earlier reports, the bitch should be maintained in the same position up to period of time varying from 5 to 20 min after AI. However, reducing the interval of elevated hindquarters to 1 min seems not affect fertility (Pinto et al., 1998). Also, feathering or stroking of the vulvar or perineal region is reported by several authors as form of stimulating the semen transport into the uterus, in an attempt to mimic the vaginal stimulation by the thrusting movements of the dog during natural mount. However, the contribution of such procedures to the exit of the technique has not been proven yet.
\n\t\t\t\tIntrauterine insemination may be performed by using non-surgical transcervical catheterisation (Linde-Forsberg, 1991; Linde-Forsberg and Forsberg, 1989, 1993; Linde-Forsberg et al., 1999) or by surgical semen deposition by laparotomy (Brittain et al., 1995; Günzel-Apel & Thiet, 1990) or laparoscopy (L.D.M Silva et al., 1995, 1996). The majority of European centers working on small animal reproduction prefer transcervical intrauterine insemination (TCI) due to reasons associated with animal welfare. However, catheterisation of uterine cervix in the bitch is a difficult procedure and demand skill and experience. The semen of lower quality, such as frozen-thawed or that collected from subfertile dogs have to be deposited intrauterine to assure satisfactory results of artificial insemination (Linde-Forsberg et al., 1999; Thomassen et al., 2006). The conception rates after intravaginal insemination with frozen-thawed semen are significantly lower when compared with the results of intrauterine insemination.
\n\t\t\t\t\tThe method of non-surgical transcervical intrauterine insemination was first time described in 1975 (Andersen, 1975). The technique has been adapted from the artificial insemination performed in foxes. Two catheters are used in this method - the outer plastic catheter and inner metal thin catheter. There are 3 sizes of the catheters, for small, medium and large breeds. The catheterisation should be made on standing animal. Sometimes there is no need for administration of sedatives, but usually a small dose of alpha-mimetic, such as medetomidine, is advisable for abdominal muscles relaxation. The outer plastic catheter should be introduced into the vagina. It should be advanced as far as is possible. In many bitches, especially those of larger breeds, the tip of the catheter passes into the cranial narrow part of vagina. However, in some smaller bitches the introduction of the outer catheter through the paracervix is difficult. It is necessary to palpate the end of the catheter and the cervix through the abdominal wall. The cervix is palpable at estrus as solid, ovoid structure. It is advisable to move the tip of the catheter ventrally towards the ventral region of abdominal wall. This procedure is helpful in palpation of the cervix. The inner metal catheter should be introduced through the plastic catheter. The cervix should be fixed between the thumb and other fingers and tilted to horizontal axis. The metal catheter is introduced into the cervical canal under the control of the position of the cervix by palpation through the abdominal wall (Andersen, 1975; Linde-Forsberg, 1991). This technique demands skill and experience. It is harder to perform uterine catheterisation in obese or nervous bitches and in giant breeds.
\n\t\t\t\t\t\tThe scandinavian method of uterine catheterization is recommended for routine insemination of bitches (Ferguson et al., 1989; Günzel-Apel 1994; Linde-Forsberg, 1991, 1993). The use of this technique of insemination is especially advisable in cases when using of semen of lower quality due to male subfertility or sperm cryopreservation. Linde-Forsberg and Forsberg (1989) obtained 83.9% and 69.3% of pregnant bitches (data corrected for the stage of estrus) after insemination with fresh and frozen-thawed semen, respectively. The litter sizes were lower by 23.3%, when frozen-thawed semen was used in comparison to fresh semen. Rota et al. (1999a) reported 25% higher pregnancy rate after intrauterine semen deposition when using scandinavian technique than after vaginal semen deposition. On the basis of analysis of 327 inseminations Linde-Forsberg et al. (1999) concluded that success rate of scandinavian method and vaginal insemination with frozen-thawed semen was 84.4% and 58.9, respectively. Niżański (2006) proved that results of vaginal insemination with frozen-thawed semen are significantly lower in comparison with fresh semen, in spite of the use of modification of the technique of vaginal semen deposition, plasma addition and adjustment of the number of spermatozoa.
\n\t\t\t\t\tIntrauterine insemination of the bitch under the visual control of endoscopic equipment was first time described by Wilson (Wilson, 1993, 2001), using a rigid endoscope – cysto-uretroscope of the length 29 cm with diagnostic external sheath. The procedure is performed on the standing animal. Uterine catheterisation is made with the use of flexible catheter introduced into the working channel of the endoscope. The endoscope is introduced into the cranial narrow part of the vagina, while a flexible catheter is introduced cranio-dorsal into the external orifice of the cervical canal under the visual control performed through the endoscope. Usually it is not necessary to administer any sedatives.
\n\t\t\t\t\t\tThe results for the intrauterine deposition of frozen-thawed semen when using this technique are quite satisfactory (Table 12). Wilson (1993), with the use of frozen semen, refers a pregnancy rate and litter size 83.3% and 7.5 puppies
Insemination technique | \n\t\t\t\t\t\t\t\t\tn | \n\t\t\t\t\t\t\t\t\tWhelping rate(n° females whelped/inseminated) | \n\t\t\t\t\t\t\t\t\tLitter size at birth (range) | \n\t\t\t\t\t\t\t\t\tLitter size at weaning (range) | \n\t\t\t\t\t\t\t\t\tPercentage of male pups/litter | \n\t\t\t\t\t\t\t\t
Intravaginal, using an infusion pipete | \n\t\t\t\t\t\t\t\t\t18 | \n\t\t\t\t\t\t\t\t\t27.8a (5/18) | \n\t\t\t\t\t\t\t\t\t3.0 ±1.2a (2-5) | \n\t\t\t\t\t\t\t\t\t2.6 ±0.9a (2-4) | \n\t\t\t\t\t\t\t\t\t51.5 ±25.9a | \n\t\t\t\t\t\t\t\t
Intrauterine, using the endoscope | \n\t\t\t\t\t\t\t\t\t16 | \n\t\t\t\t\t\t\t\t\t68.7b (11/16) | \n\t\t\t\t\t\t\t\t\t4.9 ±1.7b (3-8) | \n\t\t\t\t\t\t\t\t\t4.6 ±1.7b (2-8) | \n\t\t\t\t\t\t\t\t\t54.7 ±18.7a | \n\t\t\t\t\t\t\t\t
Different superscripts in the same column indicate significant difference (p<0.05)
Whelping rate and mean litter size in bitches intravaginally or intrauterine vaginoscopically inseminated with frozen-thawed semen. AI were performed at 4th and 6th days of oestrous cycle.
Currently a large variety of rigid endoscopes of different brands is available. The most suitable are those of the length 30-35 cm with 30° or 0° view angle and the diameter of 3 mm or similar. Some companies market the equipment dedicated specifically for artificial insemination in bitches. The optics of endoscope should be used with outer sheath equipped with working channel allowing for introduction of flexible catheter of the diameter of 5-7 French gauge. The mechanism of Albaran is not useful in the endoscope for artificial insemination of bitches, as it is not practical and the diameter of the outer sheath becomes too large when it is attached.
\n\t\t\t\t\t\tThe catheterization may be problematic in both toy and giant breeds. In the first case, the diameter of the equipment may be too large for introduction of the tip of optics near the uterine cervix, impairing visualization of external cervical orifice. In contrast, in giant breeds the length of the vagina may exclude the possibility to access the region of the cervix. One additional problem may be associated with the presence of the blood or mucus within the vagina. Sometimes the fluid present in the genital tract may cover the terminal lens of the endoscope making the observation impossible. Withdrawal the optics and cleaning of the lens is necessary in such cases. For better visualization of the vagina, especially within the cranial narrow part, insuflation with CO2 made with insuflator or even with rubber bulb is advisable. Therefore the view of tissues is focused, sharper and clearer, which makes the procedure easier. The time necessary for catheterization is variable. It depends mainly on the skill of the operator and on the anatomical features of the vagina, and varies usually between 0.5-3 minutes, but it may be longer. In nervous bitches the administration of small doses of sedatives, such medetomidine, is advisable, although in not too high doses, as the catheterisation should be done on standing bitch.
\n\t\t\t\t\t\tCurrently the vaginoscopic method of intrauterine insemination appears to be advantageous and useful technique of semen deposition in the uterine lumen in bitches. The technique demands skill but it is practical and quick to perform for experienced operators. The visual control of introduction of the catheter into the uterus is the important advantage of the technique. The observation of the moments of semen deposition and control if there is no semen backflow is therefore possible. Moreover, it allows also uterine sampling when a female is suspected of infertility due to uterine disease (Thomassen & Farstad, 2009). For these reasons this method of intrauterine insemination is becoming more popular.
\n\t\t\t\t\tSurgical insemination technique have been proposed once for frozen semen or when the bitch presents an anatomical obstruction that prevents the insertion of the catheter or endoscope. Both the laparoscopic approach and the laparotomy requires anaesthesia and good surgical skills. The semen is introduced into the uterus by puncture of uterine wall or incision with a scapel and passage of a tom cat catheter (Farstad, 2010; Thomassen & Farstad, 2009). However, in such methods, semen deposition is performed only once.
\n\t\t\t\t\tSome restrictions may exists to application of this method in different countries, that may compromise the registration of litters obtained without fulfilment of the legal requirements, such as previous evaluation of the situation or previous authorization of the local Kennel Club for the procedure. Furthermore, some ethical constraints have been raised regarding the use of surgical techniques for AI in dogs. Surgery is an invasive procedure, so it is unlikely to carry it out in the best interest of the animal, and the possibility of transmission of an undesirable trait in a particular animal genetic line should be kept in mind.
\n\t\t\t\tBefore implementing canine artificial insemination the owner and the clinician should be aware of the national or international regulations on semen import, if applicable, and of the local Kennel Club requirements respecting the use of canine AI and litter registration. In addition, procedures may differ between the use of national or imported semen. Consequently, attention should be paid well in advance to this matter.
\n\t\t\tIn the absence of a specific national regulation, most Kennel Clubs follows FCI (
Besides regulations on performing AI to a bitch, attention must be paid to issues concerning the semen collection and use. Besides the use of a recognisable male, with a certified pedigree, particular requirements may exist from national Kennel Clubs or the Official Agriculture entities, which may vary for chilled and frozen semen. In some situations the permit to import dog semen is required, which may or may not need to be accompanied by a DNA sample, and a health certificate that may include blood testing against the most important infectious or congenital canine diseases. Awareness of the latest official requirements is essential when considering semen international shipment. Additional information on the shipment regulations may be obtained throught the references Linde Forsberg, 2001, 2005b.
\n\t\tDemands for canine artificial insemination is growing worldwile together with an increase request for semen preservation in sperm banks. Furthermore, a tendency exists to increase the demands for the use of frozen/thawed semen over fresh semen AI, as part of breeding tools for genetic improvement. Nowadays is possible to achieve adequate whelping rates and litter sizes regardless of the type of semen used, as long as proper timing of AI and proper semen deposition are used. Client education and technical councelling must complete the AI services to be offered by specialized practicioners, in particular when breeding a problematic bitch.
\n\t\tThe authors are greatly indebted to Dr Malgorzata Ochota for her valuable cooperation and thoughtful comments during language revision of this manuscript.
\n\t\tTo improve the quality and quantity of an organism, both prokaryotes and eukaryotes, it can be approached by molecular character enhancement through the insertion of interest genes or superior genes into the cells of the desired organism. The process of genetic transformation of an organism involves the isolation and identification of the gene of interest, the technique of cloning the gene on a plasmid vector until the process of transferring the gene to the target organism’s cell. One of the important genes in the growth of organisms is the homeobox gene, which is a gene that regulates the growth and development of organisms in a very early stage. Homeobox genes were first discovered in the
In plants, overexpression of the homeobox gene at an early stage of growth will activate the formation of apical buds from apical meristems that will produce shoots. The addition of exogenous cytokinin and auxin growth regulators will activate the homeobox genes to induce cell division genes that in turn will produce somatic embryos. Theoretically, each somatic cell can grow and transform itself into somatic embryos, therefore it can produce plant seeds in large quantities and uniform phenotypic characters. This is very profitable for agriculture and industry, especially for the mass production of identical plant seeds using tissue culture techniques.
In the model plant,
Gene | Function | Organism | Ref |
---|---|---|---|
Homologous with Kn1 ( | [3, 4] | ||
Homologous with Kn1 (Zea mays). Affect the design of internodes resulting in stunted plants | [5] | ||
Morphogenesis | [6] | ||
Phyllotactic pattern and stem cell fate | [7] | ||
Accumulation of anthocyanin and in root development | [8] | ||
Mediates plant morphogenesis for light signals response | [9] | ||
MDH1 | Homologous | [10] | |
Controls plant architecture by locally restricting environmental responses | [11] | ||
Effects seed oil content in | [12] | ||
have species-specific roles in embryo and inflorescence development | [13] | ||
Regular | [14] | ||
The leaves morphological states | Tomato | [15] | |
Response to treatment for abscisic acid (ABA) | [16] | ||
Involved in Meristem Activity and Organ Separation | [17] | ||
Regulates the photoperiod sensitivity in | [18] | ||
Transcriptional activator | [19] | ||
Play a role in the embryo protoderm identity specification, organize of the primary root primordium or the L1 cell layer maintenance in the shoot apical meristem | [20] | ||
Master regulator for the gametophyte-to-sporophyte transition | [21] | ||
Athb-2 | Changes in light quality perceived by a novel phytochrome regulate plant development | Green plants | [22] |
Regulate leaf and flower morphology, accompanied by a decrease in the content of the active gibberellin | [14] |
The homeobox gene in plants.
Transient expression become a powerful tool in functional genomics study for detecting gene expression in a short time and the inserted gene do not integrate into the plant genome. A transient expression system has been developed in planta using different cells or tissues, including protoplast, onion cells, and tobacco (
Plant materials | Transformation methods | Purposes/Functions | Refs. |
---|---|---|---|
Protoplast | PEG | Subcellular localization of proteins | [24, 25] |
Agroinfiltration | Metabolite’s production (protein, secondary metabolite, etc.) | [26] | |
Onion cells | Particle bombardment | Subcellular localization of proteins | [27] |
Transient expression system and its purposes in planta.
The advantages of the transient expression system compared to stable expression are that it does not require regeneration of transformed cells, does not affect the stability of the host genome, and is independent of the effect of T-DNA integration site position [28]. Protoplast transfection can be performed using a variety of procedures commonly used for the transfection of animal cell cultures. The procedures that are often used to insert DNA into protoplasts are polyethylene glycol (PEG) and electroporation [29].
Polyethylene glycol (PEG)-mediated transformation plant cells can be transformed through certain chemicals, namely PEG (polyethylene glycol). PEG is an oligomer or hydrophilic polymer synthesized from ethylene oxide, containing repeating units of -(O-CH2-CH2)-. Polyethylene oxide (PEO) is another name for PEG. Typically, ethylene oxide macromolecules with a molecular weight of less than 20,000 g/mol are called PEGs, while macromolecules with values above 20,000 g/molar are called PEOs [29]. PEG is soluble in acetonitrile, benzene, water, ethanol, and dichloromethane, while it is insoluble in diethyl ether and hexane (Figures 1 and 2).
Agroinfiltration in tobacco (
Transient expression in onion cell and protoplast for determining the subcellular localization of the protein. (a) Subcellular localization of OsKAN1-GFP fusion protein in the nucleus of onion cell transformed using particle bombardment [
PEG is available in various structures, such as branched, stellar, and comb-like macromolecules. PEG can bind various reactive functional groups to the PEG polymer site. Homo and heterobifunctional PEG derivatives are particularly suitable as agents or spacers of two chemical entities, whereas mono-functional PEGs prevent linking reactions that can affect the PEGylation of certain compounds with bifunctional PEGs. PEGylation is an interesting process in which PEG is bound to other molecules [31, 32].
PEG was used to increase DNA uptake into the protoplast during transfection. Very high concentrations of PEG can reduce transfection efficiency because it is toxic to protoplasts [33]. PEG-mediated DNA uptake is a direct gene transfer method that utilizes the interaction between PEG, naked DNA, salts, and protoplast membranes to influence the transport of DNA into the cytoplasm. The advantage of PEG-mediated transformation is that it does not require special equipment and can be carried out in the laboratory under sterile conditions [34]. Compared to
Particle bombardment particles are coated in DNA and can penetrate plant cells without killing the plant cells themselves. Previous experiments have shown that particle bombardment has been successfully used to insert DNA into rice callus and seedlings grown in dark conditions but has the disadvantage of low efficiency and reliance on expensive equipment [36].
To understand the mechanism of how the transformation and expression of recombinant protein in a prokaryotic system,
Ribonuclease III (RNase III) is an enzyme that specifically cleaves double-stranded RNA [30, 37, 38, 39, 40]. RNase III has an important role in both the RNA transcript maturation and decay of diverse cellular and viral RNA. A primary function of RNase III, however, is the maturation of ribosomal RNA (rRNA) [30, 37, 38, 40, 41]. RNase III has been known to be widely distributed across the living kingdom of life, from bacteria to higher eukaryotes. RNase III family has common features in their molecular organization, by which it consists of catalytic domain with the common feature of HNERLFGDS located at the N-terminus and double-stranded binding domain (dsRBD) that located on their C-terminus [39]. RNase III exhibited enzymatically active in homodimeric form, by which each monomer has its catalytic mechanism and therefore the cleavage product of the RNase III exhibits a very regular length of short double-stranded RNA [39]. By such properties, RNase III can be manipulated to produce short dsRNA that can be implemented for the RNA interference technology in combination with Argonaute, Drosha, and Dicer [42]. Therefore, the production of recombinant RNase III is necessary from the scientific and technological point of view.
Production of recombinant proteins could be done in either bacterial or mammalian cells as a host. The choice of the host to produce recombinant protein may be the subject of proteins of interest. It depends on whether further processing of the proteins of interest is necessary or not. However, the bacterial cell is the most prominent host for recombinant protein production.
This report will discuss the production of recombinant RNase III from a psychrotrophic bacterium,
Protein adaptation in such low temperatures requires a strategy that is not commonly found in mesophilic, for example, psychrophilic proteins must be flexible enough to avoid the problem in protein folding and to perform the optimum catalytic activity if it is an enzyme. Therefore, the production of psychrophilic protein would be interesting due to their properties to adapt to such low temperatures. Although the production of recombinant protein in bacterial host seems to be straightforward, several difficulties that arise and how to solve the problems during the production of recombinant psychrophilic protein will be discussed.
To localize the
Molecular organization of
To overexpress the recombinant psychrophilic RNase III from
The result showed that the recombinant psychrophilic RNase III was accumulated in inclusion body form, although the overproduction was shifted at 20°C (Figure 4). Several attempts have been implemented to improve the solubility of recombinant psychrophilic RNase III in the
SDS-PAGE of recombinant psychrophilic RNase III overproduced in
Another strategy that has been carried out to improve the recombinant psychrophilic RNase III was by co-expression with the chaperone or chaperone-like proteins. Chaperon is a protein that functions for assisting another protein folding. Two types of assisting folding proteins used were GroEL-ES from
Psychrophilic enzymes have unique properties in their folding and activity. Expression of such recombinant psychrophilic enzymes in mesophilic host generally produces misfolding recombinant protein represented by the inclusion bodies formation. Overexpression of recombinant psychrophilic RNase III in
The yeast
Yeast cells are protected by a thick cell wall, a potential barrier to DNA invasion. Removing the cell wall to create protoplasts or spheroplasts increases the chances of genetic transformation. Reseachers adopted this method was adopted and widespread used by these researchers, but some changes have since been have been reported to improve efficiency. This method is based on the technique described initially for protoplast fusion yeast. Yeast cells are recovered in the late stage of growth, the cell wall is weakened with a reducing agent such as mercaptoethanol, and the wall is removed by incubation with an enzyme such as glucanase. Various formulations, such as glucanase enzyme and actinomycete extract have been successfully used. Spheroplasts were then carefully washed with an osmotically equivalent solution of the free buffer and suspended in a solution-containing polyethylene glycol (PEG) and CaCl2 [48]. DNA was added at this stage. For cells to divide, the walls need to be rebuilt. This case requires the cells to be placed in osmotically stabilized agar.
Both plasmid vectors and chromosomal integration are widely used to introduce genes and control copy numbers into
Class | Sub Class | Gene | Function | Organism | Ref |
---|---|---|---|---|---|
ANTP | EuHox | Essential for normal T lymphocyte and activated natural killer cell function | [55, 56] | ||
SuperHox | Essential for the development of the pancreas | Zebrafish | [55] | ||
ParaHox | Regulate LGE patterning but oppositely control the balance between proliferation and differentiation in the neuronal progenitor pool. | [55, 57] | |||
NK cluster | Essential for normal heart morphogenesis | [55, 58] | |||
Regulate postnatal myogenesis, including muscle maintenance during aging and regeneration of acute and chronic muscle injury. | [55, 59] | ||||
PRD | PAX | Regulate the development of the animal olfactory system | Mouse | [55, 60] | |
Involved in the development of the eye structures of | [55, 61] | ||||
Expressed in the presumptive midbrain at early developmental stages, and the hindbrain at later stages, with exclusion from the MHB | [55, 62] | ||||
LIM | Encoding gene regulation during the pituitary gland, eye, and pancreas, organs assembly that was presumably not present in the common ancestor of vertebrates. | [55] | |||
TALE | Plays a critical role in tendon differentiation by regulating type I collagen production in tendon cells. | [55, 63] | |||
POU | Essential for the active maintenance of the differentiated state of a neuron across phylogeny. | [55, 64] | |||
SINE | Play an essential role in retinal development and influence that these proteins have on cell proliferation and growth | [55, 65] | |||
CUT | Developing kidney with expression restricted to the nephrogenic zone. | [55, 66] | |||
ZF | Specification of individual anterior neural precursors and promotes the expression of | [55, 67] | |||
HNF | The inducible protective mechanism that inhibits LPS-induced ROS production and inflammation in EA.hy926 cells by the subsequent inhibition of redox-sensitive NF-κB and MAPK activation. | [55, 68] | |||
CERS | Synthesize ceramides containing mainly C20–C26 fatty acids, with little or no synthesis of C16- and C18-ceramides | [55, 69] | |||
PROSS | Expressed in the developing CNS, lens-secreting cone cells of the eye, and midgut. In the mouse, Prox 1 is expressed in many of the same tissues. Young neurons of the subventricular region of the CNS, developing eye lens, and pancreas. Expression is also detected in the developing liver and heart, as well as transiently in the skeletal muscles | Mouse, | [55, 70] |
Homeobox gene in animal.
The development of a vector system for gene transformation in animal cells is under consideration [71]. These vectors are required in biotechnology to synthesize recombinant proteins from genes that are not correctly expressed when cloned in
Currently, gene cloning in mammals is performed for one of three reasons: (1) To produce recombinant proteins in mammalian cell culture and related farming techniques. Milk. (2) In gene therapy, human cells are manipulated to treat diseases. (3) Achieve gene knockout, an important technique used to determine the function of unknown genes. These experiments are usually performed on rodents, such as mice. Viruses as a mammalian clone vector have been known to be the key to cloning mammals for many years. The first cloning experiment with mammalian cells was performed in 1970 using a vector-based on Simian virus 40 (SV40) [72, 73]. The virus can infect several mammalian species following a lysogenic cycle in some hosts and others. SV40 has the same problem as e and has a calicivirus embedded in it. This is because packaging restrictions limit the amount of new DNA inserted into the genome. Therefore, cloning with the SV40 requires replacing one or more of the existing genes with DNA to clone. The original experiment replaced the late gene region segment, but early gene replacement was also an option [73]. However, the discovery of CRISPR/Cas which is based on cloning technology is one of the essential techniques in gene therapy [74].
Genes are the universal language that controls the nature of all living things, shared homology among organisms. It is always interesting to reveal the evolution of cloning and gene expression in plant, bacteria, and animal cells. Therefore, with the discovery of genetic engineering, possible to exchange good genetic traits which beneficial for human life. In conclusion, genetic transformation is a genetic engineering technique that can be used to understand the function of a gene or several genes in various events in the life of an organism, both prokaryotes and eukaryotes, so that genetic transformation is carried out for two kinds of purposes, namely scientific purposes to determine the function of certain genes in an organism, and economic goals to improve the quality and productivity of an organism to increase the economic value of an organism. In the future, genetic engineering on prokaryotes and eukaryotes perspective can be used for various purposes in the fields of medicine, agriculture, horticulture, forestry, and food.
We thank Badan Penerbit dan Publikasi Universitas Gadjah Mada (BPP UGM) for supporting this publication.
The authors declare no conflict of interest.
Cauliflower Mozaic Virus 35S promoter a part of RNA-induced silencing complex (RISC), plays a central role in RNA silencing processes brassinosteroid Interacting protein 116b human RNase III a class III of RNase III double-strand binding domain double-strand RNA era protein-encoding gene peptidyl-prolyl isomerase protein, a chaperone-like protein from psychrophilic bacterium Shewanella sp. SIB1 chaperonin isopropyl thio-b-D-galactopyranoside signal peptidase encoding gene RNA silencing suppressor p19 bacterial expression vector bacterial expression vector bacterial expression vector ribonuclease III ribonuclease III encoding gene sodium dodecyl sulfate-polyacrylamide gel electrophoresis riboniclease III encoding gene from Shewanella sp. SIB1 transfer DNA
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",metaTitle:"Team",metaDescription:"Advancing discovery in Open Access for the scientists by the scientist",metaKeywords:null,canonicalURL:"page/team",contentRaw:'[{"type":"htmlEditorComponent","content":"Our business values are based on those any scientist applies to their research. We have created a culture of respect and collaboration within a relaxed, friendly and progressive atmosphere, while maintaining academic rigour.
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\\n\\nDr Alex Lazinica
\\n\\nAlex Lazinica is co-founder and Board member of IntechOpen. After obtaining a Master's degree in Mechanical Engineering, he continued his Ph.D. in Robotics at the Vienna University of Technology. There, he worked as a robotics researcher with the university's Intelligent Manufacturing Systems Group, as well as a guest researcher at various European universities, including the Swiss Federal Institute of Technology Lausanne (EPFL). During this time he published more than 20 scientific papers, gave presentations, served as a reviewer for major robotic journals and conferences and, most importantly, co-founded and built the International Journal of Advanced Robotic Systems, the world's first Open Access journal in the field of robotics. Starting this journal was a pivotal point in his career since it proved to be the pathway to the foundation of IntechOpen with its focus on addressing academic researchers’ needs. Alex personifies many of IntechOpen´s key values, including the commitment to developing mutual trust, openness, and a spirit of entrepreneurialism. Today, his focus is on defining the growth and development strategy for the company.
\\n"}]'},components:[{type:"htmlEditorComponent",content:"Our business values are based on those any scientist applies to their research. We have created a culture of respect and collaboration within a relaxed, friendly and progressive atmosphere, while maintaining academic rigour.
\n\nCo-founded by Alex Lazinica and Vedran Kordic: “We are passionate about the advancement of science. As Ph.D. researchers in Vienna, we found it difficult to access the scholarly research we needed. We created IntechOpen with the specific aim of putting the academic needs of the global research community before the business interests of publishers. Our Team is now a global one and includes highly-renowned scientists and publishers, as well as experts in disseminating your research.”
\n\nBut, one thing we have in common is -- we are all scientists at heart!
\n\nSara Uhac, COO
\n\nSara Uhac was appointed Managing Director of IntechOpen at the beginning of 2014. She directs and controls the company’s operations. Sara joined IntechOpen in 2010 as Head of Journal Publishing, a new strategically underdeveloped department at that time. After obtaining a Master's degree in Media Management, she completed her Ph.D. at the University of Lugano, Switzerland. She holds a BA in Financial Market Management from the Bocconi University in Milan, Italy, where she started her career in the American publishing house Condé Nast and further collaborated with the UK-based publishing company Time Out. Sara was awarded a professional degree in Publishing from Yale University (2012). She is a member of the professional branch association of "Publishers, Designers and Graphic Artists" at the Croatian Chamber of Commerce.
\n\nAdrian Assad De Marco
\n\nAdrian Assad De Marco joined the company as a Director in 2017. With his extensive experience in management, acquired while working for regional and global leaders, he took over direction and control of all the company's publishing processes. Adrian holds a degree in Economy and Management from the University of Zagreb, School of Economics, Croatia. A former sportsman, he continually strives to develop his skills through professional courses and specializations such as NLP (Neuro-linguistic programming).
\n\nDr Alex Lazinica
\n\nAlex Lazinica is co-founder and Board member of IntechOpen. After obtaining a Master's degree in Mechanical Engineering, he continued his Ph.D. in Robotics at the Vienna University of Technology. There, he worked as a robotics researcher with the university's Intelligent Manufacturing Systems Group, as well as a guest researcher at various European universities, including the Swiss Federal Institute of Technology Lausanne (EPFL). During this time he published more than 20 scientific papers, gave presentations, served as a reviewer for major robotic journals and conferences and, most importantly, co-founded and built the International Journal of Advanced Robotic Systems, the world's first Open Access journal in the field of robotics. Starting this journal was a pivotal point in his career since it proved to be the pathway to the foundation of IntechOpen with its focus on addressing academic researchers’ needs. Alex personifies many of IntechOpen´s key values, including the commitment to developing mutual trust, openness, and a spirit of entrepreneurialism. Today, his focus is on defining the growth and development strategy for the company.
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It represents the integration of tools already used in the past (big data, cloud, robot, 3D printing, simulation, etc.) that are now connected into a global network by transmitting digital data. The implementation of this new paradigm represents a huge change for companies, which are faced with big investments. In order to benefit from the opportunities offered by the smart revolution, companies must have the prerequisites needed to withstand changes generated by “smart” system. In addition, new workers who face the world of work 4.0 must have new skills in automation, digitization, and information technology, without forgetting soft skills. 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Therefore, this chapter deals with the philosophical systems and paradigms of scientific research, the epistemology, evaluating understanding and application of various theories and practices used in the scientific research. The key components of the scientific research paradigm are highlighted. Theories on the basis of which this research was focused on identification of the level of development of the management culture in order to implement corporate social responsibility are identified, and the stages of its implementation are described.",book:{id:"5791",slug:"management-culture-and-corporate-social-responsibility",title:"Management Culture and Corporate Social Responsibility",fullTitle:"Management Culture and Corporate Social Responsibility"},signatures:"Pranas Žukauskas, Jolita Vveinhardt and Regina Andriukaitienė",authors:[{id:"179629",title:"Prof.",name:"Jolita",middleName:null,surname:"Vveinhardt",slug:"jolita-vveinhardt",fullName:"Jolita Vveinhardt"}]},{id:"74550",title:"School Conflicts: Causes and Management Strategies in Classroom Relationships",slug:"school-conflicts-causes-and-management-strategies-in-classroom-relationships",totalDownloads:2204,totalCrossrefCites:1,totalDimensionsCites:10,abstract:"Conflicts cannot cease to exist, as they are intrinsic to human beings, forming an integral part of their moral and emotional growth. Likewise, they exist in all schools. The school is inserted in a space where the conflict manifests itself daily and assumes relevance, being the result of the multiple interpersonal relationships that occur in the school context. Thus, conflict is part of school life, which implies that teachers must have the skills to manage conflict constructively. Recognizing the diversity of school conflicts, this chapter aimed to present its causes, highlighting the main ones in the classroom, in the teacher-student relationship. It is important to conflict face and resolve it with skills to manage it properly and constructively, establishing cooperative relationships, and producing integrative solutions. Harmony and appreciation should coexist in a classroom environment and conflict should not interfere, negatively, in the teaching and learning process. This bibliography review underscore the need for during the teachers’ initial training the conflict management skills development.",book:{id:"7827",slug:null,title:"Interpersonal Relationships",fullTitle:"Interpersonal Relationships"},signatures:"Sabina Valente, Abílio Afonso Lourenço and Zsolt Németh",authors:null},{id:"58969",title:"Corruption, Causes and Consequences",slug:"corruption-causes-and-consequences",totalDownloads:27589,totalCrossrefCites:11,totalDimensionsCites:13,abstract:"Corruption is a constant in the society and occurs in all civilizations; however, it has only been in the past 20 years that this phenomenon has begun being seriously explored. It has many different shapes as well as many various effects, both on the economy and the society at large. Among the most common causes of corruption are the political and economic environment, professional ethics and morality and, of course, habits, customs, tradition and demography. Its effects on the economy (and also on the wider society) are well researched, yet still not completely. Corruption thus inhibits economic growth and affects business operations, employment and investments. It also reduces tax revenue and the effectiveness of various financial assistance programs. The wider society is influenced by a high degree of corruption in terms of lowering of trust in the law and the rule of law, education and consequently the quality of life (access to infrastructure, health care). There also does not exist an unambiguous answer as to how to deal with corruption. Something that works in one country or in one region will not necessarily be successful in another. This chapter tries to answer at least a few questions about corruption and the causes for it, its consequences and how to deal with it successfully.",book:{id:"6487",slug:"trade-and-global-market",title:"Trade and Global Market",fullTitle:"Trade and Global Market"},signatures:"Štefan Šumah",authors:[{id:"228073",title:"Mr.",name:"Stefan",middleName:null,surname:"Sumah",slug:"stefan-sumah",fullName:"Stefan Sumah"}]},{id:"55499",title:"Human Resources Management in Nonprofit Organizations: A Case Study of Istanbul Foundation for Culture and Arts",slug:"human-resources-management-in-nonprofit-organizations-a-case-study-of-istanbul-foundation-for-cultur",totalDownloads:2294,totalCrossrefCites:0,totalDimensionsCites:0,abstract:"The aim of this study is to investigate the efficiency and importance of human resources management in nonprofit organizations. The understanding was included to the literature as personnel management at the beginning of the twentieth century and it turned into an approach as human resources management in the 1980s. It could be observed that many organizations, which deem the human as the most critical stakeholder, adopt a traditional way of personnel management in operating human resources. The employees play a key role in the success of an organization. For this reason, subjects such as recruitment, training, development, career management, performance appraisal, occupational health, and safety are the fundamental functions of human resources management. The study examines to what extent these roles are evaluated through a case study. The subject matter of the study is the most powerful culture and art foundation in Turkey. Compared to many other nonprofit organizations, the foundation actively performs a variety of services within a year worldwide. The fact that the total number of employees might rise up to 800, including the field personnel, indicates the need of a good functioning human resources management. The human resources practices of the foundation are examined and evaluated within that scope.",book:{id:"5826",slug:"issues-of-human-resource-management",title:"Issues of Human Resource Management",fullTitle:"Issues of Human Resource Management"},signatures:"Beste Gökçe Parsehyan",authors:[{id:"189113",title:"Dr.",name:"Beste",middleName:null,surname:"Gokce Parsehyan",slug:"beste-gokce-parsehyan",fullName:"Beste Gokce Parsehyan"}]},{id:"59152",title:"Marketing Strategies for the Social Good",slug:"marketing-strategies-for-the-social-good",totalDownloads:1594,totalCrossrefCites:1,totalDimensionsCites:1,abstract:"Social network sites (SNS) have proven to be a good environment to promote and sell goods and services, but marketing is more than creating commercial strategies. Social marketing strategies can also be used to promote behavioral change and help individuals transform their lives, achieve well-being, and adopt prosocial behaviors. In this chapter, we seek to analyze with a netnographic study, how SNS are being employed by nonprofits and nongovernment organizations (NGOs) to enable citizens and consumers to participate in different programs and activities that promote social transformation and well-being. A particular interest is to identify how organizations are using behavioral economic tactics to nudge individuals and motivate them to engage in prosocial actions. By providing an understanding on how SNS can provide an adequate environment for the design of social marketing strategies, we believe our work has practical implications both for academicians and marketers who want to contribute in the transformation of consumer behavior and the achievement of well-being and social change.",book:{id:"6583",slug:"marketing",title:"Marketing",fullTitle:"Marketing"},signatures:"Alicia De La Pena",authors:[{id:"196878",title:"Dr.",name:"Alicia",middleName:null,surname:"De La Pena",slug:"alicia-de-la-pena",fullName:"Alicia De La Pena"}]}],onlineFirstChaptersFilter:{topicId:"4",limit:6,offset:0},onlineFirstChaptersCollection:[{id:"82425",title:"Financial Reporting and Analysis of Tesla Green Technology in the United States Market",slug:"financial-reporting-and-analysis-of-tesla-green-technology-in-the-united-states-market",totalDownloads:0,totalDimensionsCites:0,doi:"10.5772/intechopen.105065",abstract:"This study aims to discuss and analyze the financial position and performance of the US Tesla green technology company in the United States. This study uses a case study approach, financial data, and website methodologies to collect and analyze the research data. The case study is Tesla, Inc., which is a US electric vehicle and clean energy company based in Austin, Texas. Tesla is a green technology company that produces and designs electric cars, battery energy storage from home to grid-scale, solar roof tiles and solar panels, and related products and services. Tesla is growing fastly by introducing new green products, and it is now one of the world’s most valuable enterprises. It has a high market capitalization of almost US$1 trillion to become the world’s most valuable automaker. This study concludes that Tesla has changed their strategy to become the most worldwide sales of purely battery electric vehicles, capturing 23% of the market and 16% of the plug-in electric battery in the market for 2020. It has also developed a significant installer of photovoltaic systems through its subsidiary Tesla Energy in the United States. One of the largest global battery energy-storage systems suppliers is Tesla Energy, with 3.99 gigawatt-hours installed in 2021.",book:{id:"11251",title:"Banking and Accounting",coverURL:"https://cdn.intechopen.com/books/images_new/11251.jpg"},signatures:"Nizar Mohammad Alsharari"},{id:"82427",title:"Our Globalization Era among Success, Obstacles and Doubts",slug:"our-globalization-era-among-success-obstacles-and-doubts",totalDownloads:0,totalDimensionsCites:0,doi:"10.5772/intechopen.105545",abstract:"In the last decades, the never-ending and unlimited expanding of both international economies and operations became globalization. Among its main features, one could recall the enormous increase of world macro-economic quantities (Gross World Product, Inter-continental Trade, FDI), as well as financial values (public debts and currency printing). The chapter tries to quantify them, by a statistical analysis of historical data (Section 1). Section 2 is dedicated to the strategic problems of firms, in particular the threats and opportunities for (inter) national firms willing to become global, and obstacles are included in Section 3. This given, it deals with the behavior of countries from the political and juridical points of view, and those ones passed form initial perplexities, distaste, or even hostility to a favorable behavior. Conclusions (Section 4) recall both the problematic alternative for globalized companies between “the world as our next door” and their social responsibilities and the similar problem for host countries, between socioeconomic advantages and protection of local workers, resources, and environment.",book:{id:"11476",title:"Globalization and Sustainability - Recent Advances, New Perspectives and Emerging Issues",coverURL:"https://cdn.intechopen.com/books/images_new/11476.jpg"},signatures:"Arnaldo Canziani, Annalisa Baldissera and Ahmad Kahwaji"},{id:"82289",title:"Consumer Culture and Abundance of Choices: Having More, Feeling Blue",slug:"consumer-culture-and-abundance-of-choices-having-more-feeling-blue",totalDownloads:1,totalDimensionsCites:0,doi:"10.5772/intechopen.105607",abstract:"The defining feature of contemporary consumer culture is the escalation of consumption opportunities and the expanding space for choice. An unbridled and unrestricted range of products is part of material prosperity, rising living standards, and emancipation of human freedoms. The growing demands for constant consumer decision-making in an increasingly opaque environment of potential targets of choice exposes consumers to the risk of procrastination, passivity, and resignation, as well as psychological discomfort. The goal here is to contribute to theories of consumer behavior in the context of the psychological experience of choice under the conditions of the accelerated quantity of consumption volumes against the backdrop of the COVID-19 pandemic. While conventional offline shopping was drastically curtailed during the coronavirus crisis, freedom of consumer choice was maintained despite many proclamations to the contrary. I seek to provide support to the claim that freedom of consumer choice was maintained and often amplified during the pandemic in the online virtual environment of digital commerce formats. Freedom of consumer choice has merely been transformed into a horizontal level of application by the relatively rapid and fluid conversion of market activities into the cyberspace of a growing number of e-stores and online supermarkets, unconstrained by the physical space of shelves and counters.",book:{id:"11581",title:"A New Era of Consumer Behavior - Beyond the Pandemic",coverURL:"https://cdn.intechopen.com/books/images_new/11581.jpg"},signatures:"Ondřej Roubal"},{id:"82405",title:"Does Board Structure Matter in CSR Spending of Commercial Banks? Empirical Evidence from an Emerging Economy",slug:"does-board-structure-matter-in-csr-spending-of-commercial-banks-empirical-evidence-from-an-emerging-",totalDownloads:2,totalDimensionsCites:0,doi:"10.5772/intechopen.105589",abstract:"This chapter examines the impact of board elements on CSR spending by private commercial banks in an emerging economy, considering Bangladesh as a case. In doing so, we collected necessary data from the annual reports of 30 commercial banks listed on the Dhaka Stock Exchange, covering the period 2007–2020. In addition, we reviewed the patterns of CSR spending by commercial banks to understand the CSR universe in Bangladesh. We adopted the OLS model with two-way clustering to measure the effects of board elements on CSR spending. Our results confirm that factors, such as independent directors and board size, have a significant and positive relationship with CSR expenditures, while board gender deters the same. Also, board meetings do not have any significant connection with CSR spending. For control variables, factors, such as firm size and leverage, tend to promote the CSR spending of commercial banks, while profitability has no such relationship. As for the sectoral distribution of CSR funds, we found that although the absolute amount of CSR expenditures by banks has increased substantially over the years, they are primarily limited to health, education, natural disasters, and humanitarian activities. These findings are expected to have significant policy implications.",book:{id:"11602",title:"Corporate Social Responsibility",coverURL:"https://cdn.intechopen.com/books/images_new/11602.jpg"},signatures:"Bishnu Kumar Adhikary and Ranjan Kumar Mitra"},{id:"82395",title:"Toward a Better Understanding of Green Human Resource Management’s Impact on Green Competitive Advantage: A Conceptual Model",slug:"toward-a-better-understanding-of-green-human-resource-management-s-impact-on-green-competitive-advan",totalDownloads:6,totalDimensionsCites:0,doi:"10.5772/intechopen.105528",abstract:"Today, green human resource management (GHRM) has become a key business strategy where HRM plays an active role in the ongoing green movement. Thus, the topic of GHRM is of growing interest among management scholars. However, despite the theoretically important role of GHRM, relatively small number of research has been discovered so far about how GHRM, in companies striving to achieve environmental sustainability, could help them gain a green competitive advantage (GCA). Thus, based on the resource-based view (RBV) arguments, the main objective of this paper is to develop a conceptual model of the relationship between GHRM and green competitive advantage through green knowledge, green values, and green commitment. This model is expected to provide a strategic map that could be utilized by the practitioners and managers so that GHRM implementation can be more effective in contributing to green competitive advantage. Overall, the present article extends knowledge on the resource-based view by contributing to the literature on GHRM and its interactions with the main assets that lead to green competitive advantage.",book:{id:"11602",title:"Corporate Social Responsibility",coverURL:"https://cdn.intechopen.com/books/images_new/11602.jpg"},signatures:"Hosna Hossari and Kaoutar Elfahli"},{id:"82394",title:"Learning by Doing Active Social Learning",slug:"learning-by-doing-active-social-learning",totalDownloads:2,totalDimensionsCites:0,doi:"10.5772/intechopen.105523",abstract:"Project-based learning and future-based pedagogy are important and effective tools for teaching and learning in the twenty-first century. They are especially suited to instilling social activism among students, which is extremely valuable in today’s multicultural society. This study examined the impact of such learning among Arab and Jewish students and teachers in Israel. Following a collaborative program on social activism, in which students from different sectors worked together via digital platforms and face-to-face encounters, the impact of the program and its pedagogical tools were examined. The program, called Living in a Multicultural Society, reflects the mosaic of different people and communities, living side by side yet separated by religion, culture, and language. Through this program, students who may not have otherwise met worked together to learn, research, and create. This study was conducted using the mixed-method approach, whereby the qualitative data were gathered via interviews, and the quantitative data were collected through questionnaires. The findings show that this project-based learning program led to significant encounters, understandings, and co-operations between different sectors, and to meaningful end-products relating to social activism. This study enhances the concept that significant pedagogical processes increase students’ motivation, in-depth learning, and outcomes.",book:{id:"11481",title:"Active Learning - Research and Practice",coverURL:"https://cdn.intechopen.com/books/images_new/11481.jpg"},signatures:"Anat Raviv"}],onlineFirstChaptersTotal:278},preDownload:{success:null,errors:{}},subscriptionForm:{success:null,errors:{}},aboutIntechopen:{},privacyPolicy:{},peerReviewing:{},howOpenAccessPublishingWithIntechopenWorks:{},sponsorshipBooks:{sponsorshipBooks:[],offset:0,limit:8,total:null},allSeries:{pteSeriesList:[{id:"14",title:"Artificial Intelligence",numberOfPublishedBooks:9,numberOfPublishedChapters:89,numberOfOpenTopics:6,numberOfUpcomingTopics:0,issn:"2633-1403",doi:"10.5772/intechopen.79920",isOpenForSubmission:!0},{id:"7",title:"Biomedical Engineering",numberOfPublishedBooks:12,numberOfPublishedChapters:104,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2631-5343",doi:"10.5772/intechopen.71985",isOpenForSubmission:!0}],lsSeriesList:[{id:"11",title:"Biochemistry",numberOfPublishedBooks:32,numberOfPublishedChapters:315,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2632-0983",doi:"10.5772/intechopen.72877",isOpenForSubmission:!0},{id:"25",title:"Environmental Sciences",numberOfPublishedBooks:1,numberOfPublishedChapters:12,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2754-6713",doi:"10.5772/intechopen.100362",isOpenForSubmission:!0},{id:"10",title:"Physiology",numberOfPublishedBooks:11,numberOfPublishedChapters:141,numberOfOpenTopics:4,numberOfUpcomingTopics:0,issn:"2631-8261",doi:"10.5772/intechopen.72796",isOpenForSubmission:!0}],hsSeriesList:[{id:"3",title:"Dentistry",numberOfPublishedBooks:8,numberOfPublishedChapters:129,numberOfOpenTopics:2,numberOfUpcomingTopics:0,issn:"2631-6218",doi:"10.5772/intechopen.71199",isOpenForSubmission:!0},{id:"6",title:"Infectious Diseases",numberOfPublishedBooks:13,numberOfPublishedChapters:113,numberOfOpenTopics:3,numberOfUpcomingTopics:1,issn:"2631-6188",doi:"10.5772/intechopen.71852",isOpenForSubmission:!0},{id:"13",title:"Veterinary Medicine and Science",numberOfPublishedBooks:11,numberOfPublishedChapters:105,numberOfOpenTopics:3,numberOfUpcomingTopics:0,issn:"2632-0517",doi:"10.5772/intechopen.73681",isOpenForSubmission:!0}],sshSeriesList:[{id:"22",title:"Business, Management and Economics",numberOfPublishedBooks:1,numberOfPublishedChapters:19,numberOfOpenTopics:2,numberOfUpcomingTopics:1,issn:"2753-894X",doi:"10.5772/intechopen.100359",isOpenForSubmission:!0},{id:"23",title:"Education and Human Development",numberOfPublishedBooks:0,numberOfPublishedChapters:5,numberOfOpenTopics:1,numberOfUpcomingTopics:1,issn:null,doi:"10.5772/intechopen.100360",isOpenForSubmission:!0},{id:"24",title:"Sustainable Development",numberOfPublishedBooks:0,numberOfPublishedChapters:15,numberOfOpenTopics:5,numberOfUpcomingTopics:0,issn:null,doi:"10.5772/intechopen.100361",isOpenForSubmission:!0}],testimonialsList:[{id:"6",text:"It is great to work with the IntechOpen to produce a worthwhile collection of research that also becomes a great educational resource and guide for future research endeavors.",author:{id:"259298",name:"Edward",surname:"Narayan",institutionString:null,profilePictureURL:"https://mts.intechopen.com/storage/users/259298/images/system/259298.jpeg",slug:"edward-narayan",institution:{id:"3",name:"University of Queensland",country:{id:null,name:"Australia"}}}},{id:"13",text:"The collaboration with and support of the technical staff of IntechOpen is fantastic. 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Saxena",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRET3QAO/Profile_Picture_2022-05-10T10:10:26.jpeg",institutionString:"King George's Medical University",institution:{name:"King George's Medical University",institutionURL:null,country:{name:"India"}}}],equalEditorOne:null,equalEditorTwo:null,equalEditorThree:null}]},subseriesFiltersForPublishedBooks:[{group:"subseries",caption:"Bacterial Infectious Diseases",value:3,count:2},{group:"subseries",caption:"Parasitic Infectious Diseases",value:5,count:4},{group:"subseries",caption:"Viral Infectious Diseases",value:6,count:7}],publicationYearFilters:[{group:"publicationYear",caption:"2022",value:2022,count:2},{group:"publicationYear",caption:"2021",value:2021,count:4},{group:"publicationYear",caption:"2020",value:2020,count:3},{group:"publicationYear",caption:"2019",value:2019,count:3},{group:"publicationYear",caption:"2018",value:2018,count:1}],authors:{paginationCount:301,paginationItems:[{id:"116250",title:"Dr.",name:"Nima",middleName:null,surname:"Rezaei",slug:"nima-rezaei",fullName:"Nima Rezaei",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/116250/images/system/116250.jpg",biography:"Professor Nima Rezaei obtained an MD from Tehran University of Medical Sciences, Iran. He also obtained an MSc in Molecular and Genetic Medicine, and a Ph.D. in Clinical Immunology and Human Genetics from the University of Sheffield, UK. He also completed a short-term fellowship in Pediatric Clinical Immunology and Bone Marrow Transplantation at Newcastle General Hospital, England. Dr. Rezaei is a Full Professor of Immunology and Vice Dean of International Affairs and Research, at the School of Medicine, Tehran University of Medical Sciences, and the co-founder and head of the Research Center for Immunodeficiencies. He is also the founding president of the Universal Scientific Education and Research Network (USERN). Dr. Rezaei has directed more than 100 research projects and has designed and participated in several international collaborative projects. He is an editor, editorial assistant, or editorial board member of more than forty international journals. He has edited more than 50 international books, presented more than 500 lectures/posters in congresses/meetings, and published more than 1,100 scientific papers in international journals.",institutionString:"Tehran University of Medical Sciences",institution:{name:"Tehran University of Medical Sciences",country:{name:"Iran"}}},{id:"180733",title:"Dr.",name:"Jean",middleName:null,surname:"Engohang-Ndong",slug:"jean-engohang-ndong",fullName:"Jean Engohang-Ndong",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/180733/images/system/180733.png",biography:"Dr. Jean Engohang-Ndong was born and raised in Gabon. After obtaining his Associate Degree of Science at the University of Science and Technology of Masuku, Gabon, he continued his education in France where he obtained his BS, MS, and Ph.D. in Medical Microbiology. He worked as a post-doctoral fellow at the Public Health Research Institute (PHRI), Newark, NJ for four years before accepting a three-year faculty position at Brigham Young University-Hawaii. Dr. Engohang-Ndong is a tenured faculty member with the academic rank of Full Professor at Kent State University, Ohio, where he teaches a wide range of biological science courses and pursues his research in medical and environmental microbiology. Recently, he expanded his research interest to epidemiology and biostatistics of chronic diseases in Gabon.",institutionString:"Kent State University",institution:{name:"Kent State University",country:{name:"United States of America"}}},{id:"188773",title:"Prof.",name:"Emmanuel",middleName:null,surname:"Drouet",slug:"emmanuel-drouet",fullName:"Emmanuel Drouet",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/188773/images/system/188773.png",biography:"Emmanuel Drouet, PharmD, is a Professor of Virology at the Faculty of Pharmacy, the University Grenoble-Alpes, France. As a head scientist at the Institute of Structural Biology in Grenoble, Dr. Drouet’s research investigates persisting viruses in humans (RNA and DNA viruses) and the balance with our host immune system. He focuses on these viruses’ effects on humans (both their impact on pathology and their symbiotic relationships in humans). He has an excellent track record in the herpesvirus field, and his group is engaged in clinical research in the field of Epstein-Barr virus diseases. He is the editor of the online Encyclopedia of Environment and he coordinates the Universal Health Coverage education program for the BioHealth Computing Schools of the European Institute of Science.",institutionString:null,institution:{name:"Grenoble Alpes University",country:{name:"France"}}},{id:"131400",title:"Prof.",name:"Alfonso J.",middleName:null,surname:"Rodriguez-Morales",slug:"alfonso-j.-rodriguez-morales",fullName:"Alfonso J. Rodriguez-Morales",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/131400/images/system/131400.png",biography:"Dr. Rodriguez-Morales is an expert in tropical and emerging diseases, particularly zoonotic and vector-borne diseases (especially arboviral diseases). He is the president of the Travel Medicine Committee of the Pan-American Infectious Diseases Association (API), as well as the president of the Colombian Association of Infectious Diseases (ACIN). He is a member of the Committee on Tropical Medicine, Zoonoses, and Travel Medicine of ACIN. He is a vice-president of the Latin American Society for Travel Medicine (SLAMVI) and a Member of the Council of the International Society for Infectious Diseases (ISID). Since 2014, he has been recognized as a Senior Researcher, at the Ministry of Science of Colombia. He is a professor at the Faculty of Medicine of the Fundacion Universitaria Autonoma de las Americas, in Pereira, Risaralda, Colombia. He is an External Professor, Master in Research on Tropical Medicine and International Health, Universitat de Barcelona, Spain. He is also a professor at the Master in Clinical Epidemiology and Biostatistics, Universidad Científica del Sur, Lima, Peru. In 2021 he has been awarded the “Raul Isturiz Award” Medal of the API. Also, in 2021, he was awarded with the “Jose Felix Patiño” Asclepius Staff Medal of the Colombian Medical College, due to his scientific contributions to COVID-19 during the pandemic. He is currently the Editor in Chief of the journal Travel Medicine and Infectious Diseases. His Scopus H index is 47 (Google Scholar H index, 68).",institutionString:"Institución Universitaria Visión de las Américas, Colombia",institution:null},{id:"332819",title:"Dr.",name:"Chukwudi Michael",middleName:"Michael",surname:"Egbuche",slug:"chukwudi-michael-egbuche",fullName:"Chukwudi Michael Egbuche",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/332819/images/14624_n.jpg",biography:"I an Dr. Chukwudi Michael Egbuche. I am a Senior Lecturer in the Department of Parasitology and Entomology, Nnamdi Azikiwe University, Awka.",institutionString:null,institution:{name:"Nnamdi Azikiwe University",country:{name:"Nigeria"}}},{id:"284232",title:"Mr.",name:"Nikunj",middleName:"U",surname:"Tandel",slug:"nikunj-tandel",fullName:"Nikunj Tandel",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/284232/images/8275_n.jpg",biography:'Mr. Nikunj Tandel has completed his Master\'s degree in Biotechnology from VIT University, India in the year of 2012. He is having 8 years of research experience especially in the field of malaria epidemiology, immunology, and nanoparticle-based drug delivery system against the infectious diseases, autoimmune disorders and cancer. He has worked for the NIH funded-International Center of Excellence in Malaria Research project "Center for the study of complex malaria in India (CSCMi)" in collaboration with New York University. The preliminary objectives of the study are to understand and develop the evidence-based tools and interventions for the control and prevention of malaria in different sites of the INDIA. Alongside, with the help of next-generation genomics study, the team has studied the antimalarial drug resistance in India. Further, he has extended his research in the development of Humanized mice for the study of liver-stage malaria and identification of molecular marker(s) for the Artemisinin resistance. At present, his research focuses on understanding the role of B cells in the activation of CD8+ T cells in malaria. Received the CSIR-SRF (Senior Research Fellow) award-2018, FIMSA (Federation of Immunological Societies of Asia-Oceania) Travel Bursary award to attend the IUIS-IIS-FIMSA Immunology course-2019',institutionString:"Nirma University",institution:{name:"Nirma University",country:{name:"India"}}},{id:"334383",title:"Ph.D.",name:"Simone",middleName:"Ulrich",surname:"Ulrich Picoli",slug:"simone-ulrich-picoli",fullName:"Simone Ulrich Picoli",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/334383/images/15919_n.jpg",biography:"Graduated in Pharmacy from Universidade Luterana do Brasil (1999), Master in Agricultural and Environmental Microbiology from Federal University of Rio Grande do Sul (2002), Specialization in Clinical Microbiology from Universidade de São Paulo, USP (2007) and PhD in Sciences in Gastroenterology and Hepatology (2012). She is currently an Adjunct Professor at Feevale University in Medicine and Biomedicine courses and a permanent professor of the Academic Master\\'s Degree in Virology. She has experience in the field of Microbiology, with an emphasis on Bacteriology, working mainly on the following topics: bacteriophages, bacterial resistance, clinical microbiology and food microbiology.",institutionString:null,institution:{name:"Universidade Feevale",country:{name:"Brazil"}}},{id:"229220",title:"Dr.",name:"Amjad",middleName:"Islam",surname:"Aqib",slug:"amjad-aqib",fullName:"Amjad Aqib",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/229220/images/system/229220.png",biography:"Dr. Amjad Islam Aqib obtained a DVM and MSc (Hons) from University of Agriculture Faisalabad (UAF), Pakistan, and a PhD from the University of Veterinary and Animal Sciences Lahore, Pakistan. Dr. Aqib joined the Department of Clinical Medicine and Surgery at UAF for one year as an assistant professor where he developed a research laboratory designated for pathogenic bacteria. Since 2018, he has been Assistant Professor/Officer in-charge, Department of Medicine, Manager Research Operations and Development-ORIC, and President One Health Club at Cholistan University of Veterinary and Animal Sciences, Bahawalpur, Pakistan. He has nearly 100 publications to his credit. His research interests include epidemiological patterns and molecular analysis of antimicrobial resistance and modulation and vaccine development against animal pathogens of public health concern.",institutionString:"Cholistan University of Veterinary and Animal Sciences",institution:null},{id:"62900",title:"Prof.",name:"Fethi",middleName:null,surname:"Derbel",slug:"fethi-derbel",fullName:"Fethi Derbel",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/62900/images/system/62900.jpeg",biography:"Professor Fethi Derbel was born in 1960 in Tunisia. He received his medical degree from the Sousse Faculty of Medicine at Sousse, University of Sousse, Tunisia. He completed his surgical residency in General Surgery at the University Hospital Farhat Hached of Sousse and was a member of the Unit of Liver Transplantation in the University of Rennes, France. He then worked in the Department of Surgery at the Sahloul University Hospital in Sousse. Professor Derbel is presently working at the Clinique les Oliviers, Sousse, Tunisia. His hospital activities are mostly concerned with laparoscopic, colorectal, pancreatic, hepatobiliary, and gastric surgery. He is also very interested in hernia surgery and performs ventral hernia repairs and inguinal hernia repairs. He has been a member of the GREPA and Tunisian Hernia Society (THS). During his residency, he managed patients suffering from diabetic foot, and he was very interested in this pathology. For this reason, he decided to coordinate a book project dealing with the diabetic foot. Professor Derbel has published many articles in journals and collaborates intensively with IntechOpen Access Publisher as an editor.",institutionString:"Clinique les Oliviers",institution:null},{id:"300144",title:"Dr.",name:"Meriem",middleName:null,surname:"Braiki",slug:"meriem-braiki",fullName:"Meriem Braiki",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/300144/images/system/300144.jpg",biography:"Dr. Meriem Braiki is a specialist in pediatric surgeon from Tunisia. She was born in 1985. She received her medical degree from the University of Medicine at Sousse, Tunisia. She achieved her surgical residency training periods in Pediatric Surgery departments at University Hospitals in Monastir, Tunis and France.\r\nShe is currently working at the Pediatric surgery department, Sidi Bouzid Hospital, Tunisia. Her hospital activities are mostly concerned with laparoscopic, parietal, urological and digestive surgery. She has published several articles in diffrent journals.",institutionString:"Sidi Bouzid Regional Hospital",institution:null},{id:"229481",title:"Dr.",name:"Erika M.",middleName:"Martins",surname:"de Carvalho",slug:"erika-m.-de-carvalho",fullName:"Erika M. de Carvalho",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/229481/images/6397_n.jpg",biography:null,institutionString:null,institution:{name:"Oswaldo Cruz Foundation",country:{name:"Brazil"}}},{id:"186537",title:"Prof.",name:"Tonay",middleName:null,surname:"Inceboz",slug:"tonay-inceboz",fullName:"Tonay Inceboz",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/186537/images/system/186537.jfif",biography:"I was graduated from Ege University of Medical Faculty (Turkey) in 1988 and completed his Med. PhD degree in Medical Parasitology at the same university. I became an Associate Professor in 2008 and Professor in 2014. I am currently working as a Professor at the Department of Medical Parasitology at Dokuz Eylul University, Izmir, Turkey.\n\nI have given many lectures, presentations in different academic meetings. I have more than 60 articles in peer-reviewed journals, 18 book chapters, 1 book editorship.\n\nMy research interests are Echinococcus granulosus, Echinococcus multilocularis (diagnosis, life cycle, in vitro and in vivo cultivation), and Trichomonas vaginalis (diagnosis, PCR, and in vitro cultivation).",institutionString:"Dokuz Eylül University",institution:{name:"Dokuz Eylül University",country:{name:"Turkey"}}},{id:"71812",title:"Prof.",name:"Hanem Fathy",middleName:"Fathy",surname:"Khater",slug:"hanem-fathy-khater",fullName:"Hanem Fathy Khater",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/71812/images/1167_n.jpg",biography:"Prof. Khater is a Professor of Parasitology at Benha University, Egypt. She studied for her doctoral degree, at the Department of Entomology, College of Agriculture, Food and Natural Resources, University of Missouri, Columbia, USA. She has completed her Ph.D. degrees in Parasitology in Egypt, from where she got the award for “the best scientific Ph.D. dissertation”. She worked at the School of Biological Sciences, Bristol, England, the UK in controlling insects of medical and veterinary importance as a grant from Newton Mosharafa, the British Council. Her research is focused on searching of pesticides against mosquitoes, house flies, lice, green bottle fly, camel nasal botfly, soft and hard ticks, mites, and the diamondback moth as well as control of several parasites using safe and natural materials to avoid drug resistances and environmental contamination.",institutionString:null,institution:{name:"Banha University",country:{name:"Egypt"}}},{id:"99780",title:"Prof.",name:"Omolade",middleName:"Olayinka",surname:"Okwa",slug:"omolade-okwa",fullName:"Omolade Okwa",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/99780/images/system/99780.jpg",biography:"Omolade Olayinka Okwa is presently a Professor of Parasitology at Lagos State University, Nigeria. She has a PhD in Parasitology (1997), an MSc in Cellular Parasitology (1992), and a BSc (Hons) Zoology (1990) all from the University of Ibadan, Nigeria. She teaches parasitology at the undergraduate and postgraduate levels. She was a recipient of a Commonwealth fellowship supported by British Council tenable at the Centre for Entomology and Parasitology (CAEP), Keele University, United Kingdom between 2004 and 2005. She was awarded an Honorary Visiting Research Fellow at the same university from 2005 to 2007. \nShe has been an external examiner to the Department of Veterinary Microbiology and Parasitology, University of Ibadan, MSc programme between 2010 and 2012. She is a member of the Nigerian Society of Experimental Biology (NISEB), Parasitology and Public Health Society of Nigeria (PPSN), Science Association of Nigeria (SAN), Zoological Society of Nigeria (ZSN), and is Vice Chairperson of the Organisation of Women in Science (OWSG), LASU chapter. She served as Head of Department of Zoology and Environmental Biology, Lagos State University from 2007 to 2010 and 2014 to 2016. She is a reviewer for several local and international journals such as Unilag Journal of Science, Libyan Journal of Medicine, Journal of Medicine and Medical Sciences, and Annual Research and Review in Science. \nShe has authored 45 scientific research publications in local and international journals, 8 scientific reviews, 4 books, and 3 book chapters, which includes the books “Malaria Parasites” and “Malaria” which are IntechOpen access publications.",institutionString:"Lagos State University",institution:{name:"Lagos State University",country:{name:"Nigeria"}}},{id:"273100",title:"Dr.",name:"Vijay",middleName:null,surname:"Gayam",slug:"vijay-gayam",fullName:"Vijay Gayam",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/273100/images/system/273100.jpeg",biography:"Dr. Vijay Bhaskar Reddy Gayam is currently practicing as an internist at Interfaith Medical Center in Brooklyn, New York, USA. He is also a Clinical Assistant Professor at the SUNY Downstate University Hospital and Adjunct Professor of Medicine at the American University of Antigua. He is a holder of an M.B.B.S. degree bestowed to him by Osmania Medical College and received his M.D. at Interfaith Medical Center. His career goals thus far have heavily focused on direct patient care, medical education, and clinical research. He currently serves in two leadership capacities; Assistant Program Director of Medicine at Interfaith Medical Center and as a Councilor for the American\r\nFederation for Medical Research. As a true academician and researcher, he has more than 50 papers indexed in international peer-reviewed journals. He has also presented numerous papers in multiple national and international scientific conferences. His areas of research interest include general internal medicine, gastroenterology and hepatology. He serves as an editor, editorial board member and reviewer for multiple international journals. His research on Hepatitis C has been very successful and has led to multiple research awards, including the 'Equity in Prevention and Treatment Award” from the New York Department of Health Viral Hepatitis Symposium (2018) and the 'Presidential Poster Award” awarded to him by the American College of Gastroenterology (2018). He was also awarded 'Outstanding Clinician in General Medicine” by Venus International Foundation for his extensive research expertise and services, perform over and above the standard expected in the advancement of healthcare, patient safety and quality of care.",institutionString:"Interfaith Medical Center",institution:{name:"Interfaith Medical Center",country:{name:"United States of America"}}},{id:"93517",title:"Dr.",name:"Clement",middleName:"Adebajo",surname:"Meseko",slug:"clement-meseko",fullName:"Clement Meseko",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/93517/images/system/93517.jpg",biography:"Dr. Clement Meseko obtained DVM and PhD degree in Veterinary Medicine and Virology respectively. He has worked for over 20 years in both private and public sectors including the academia, contributing to knowledge and control of infectious disease. Through the application of epidemiological skill, classical and molecular virological skills, he investigates viruses of economic and public health importance for the mitigation of the negative impact on people, animal and the environment in the context of Onehealth. \r\nDr. Meseko’s field experience on animal and zoonotic diseases and pathogen dynamics at the human-animal interface over the years shaped his carrier in research and scientific inquiries. He has been part of the investigation of Highly Pathogenic Avian Influenza incursions in sub Saharan Africa and monitors swine Influenza (Pandemic influenza Virus) agro-ecology and potential for interspecies transmission. He has authored and reviewed a number of journal articles and book chapters.",institutionString:"National Veterinary Research Institute",institution:{name:"National Veterinary Research Institute",country:{name:"Nigeria"}}},{id:"158026",title:"Prof.",name:"Shailendra K.",middleName:null,surname:"Saxena",slug:"shailendra-k.-saxena",fullName:"Shailendra K. Saxena",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRET3QAO/Profile_Picture_2022-05-10T10:10:26.jpeg",biography:"Professor Dr. Shailendra K. Saxena is a vice dean and professor at King George's Medical University, Lucknow, India. His research interests involve understanding the molecular mechanisms of host defense during human viral infections and developing new predictive, preventive, and therapeutic strategies for them using Japanese encephalitis virus (JEV), HIV, and emerging viruses as a model via stem cell and cell culture technologies. His research work has been published in various high-impact factor journals (Science, PNAS, Nature Medicine) with a high number of citations. He has received many awards and honors in India and abroad including various Young Scientist Awards, BBSRC India Partnering Award, and Dr. JC Bose National Award of Department of Biotechnology, Min. of Science and Technology, Govt. of India. Dr. Saxena is a fellow of various international societies/academies including the Royal College of Pathologists, United Kingdom; Royal Society of Medicine, London; Royal Society of Biology, United Kingdom; Royal Society of Chemistry, London; and Academy of Translational Medicine Professionals, Austria. He was named a Global Leader in Science by The Scientist. He is also an international opinion leader/expert in vaccination for Japanese encephalitis by IPIC (UK).",institutionString:"King George's Medical University",institution:{name:"King George's Medical University",country:{name:"India"}}},{id:"94928",title:"Dr.",name:"Takuo",middleName:null,surname:"Mizukami",slug:"takuo-mizukami",fullName:"Takuo Mizukami",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/94928/images/6402_n.jpg",biography:null,institutionString:null,institution:{name:"National Institute of Infectious Diseases",country:{name:"Japan"}}},{id:"233433",title:"Dr.",name:"Yulia",middleName:null,surname:"Desheva",slug:"yulia-desheva",fullName:"Yulia Desheva",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/233433/images/system/233433.png",biography:"Dr. Yulia Desheva is a leading researcher at the Institute of Experimental Medicine, St. Petersburg, Russia. She is a professor in the Stomatology Faculty, St. Petersburg State University. She has expertise in the development and evaluation of a wide range of live mucosal vaccines against influenza and bacterial complications. Her research interests include immunity against influenza and COVID-19 and the development of immunization schemes for high-risk individuals.",institutionString:'Federal State Budgetary Scientific Institution "Institute of Experimental Medicine"',institution:null},{id:"238958",title:"Mr.",name:"Atamjit",middleName:null,surname:"Singh",slug:"atamjit-singh",fullName:"Atamjit Singh",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/238958/images/6575_n.jpg",biography:null,institutionString:null,institution:null},{id:"333753",title:"Dr.",name:"Rais",middleName:null,surname:"Ahmed",slug:"rais-ahmed",fullName:"Rais Ahmed",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/333753/images/20168_n.jpg",biography:null,institutionString:null,institution:null},{id:"252058",title:"M.Sc.",name:"Juan",middleName:null,surname:"Sulca",slug:"juan-sulca",fullName:"Juan Sulca",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/252058/images/12834_n.jpg",biography:null,institutionString:null,institution:null},{id:"191392",title:"Dr.",name:"Marimuthu",middleName:null,surname:"Govindarajan",slug:"marimuthu-govindarajan",fullName:"Marimuthu Govindarajan",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/191392/images/5828_n.jpg",biography:"Dr. M. Govindarajan completed his BSc degree in Zoology at Government Arts College (Autonomous), Kumbakonam, and MSc, MPhil, and PhD degrees at Annamalai University, Annamalai Nagar, Tamil Nadu, India. He is serving as an assistant professor at the Department of Zoology, Annamalai University. His research interests include isolation, identification, and characterization of biologically active molecules from plants and microbes. He has identified more than 20 pure compounds with high mosquitocidal activity and also conducted high-quality research on photochemistry and nanosynthesis. He has published more than 150 studies in journals with impact factor and 2 books in Lambert Academic Publishing, Germany. He serves as an editorial board member in various national and international scientific journals.",institutionString:null,institution:null},{id:"274660",title:"Dr.",name:"Damodar",middleName:null,surname:"Paudel",slug:"damodar-paudel",fullName:"Damodar Paudel",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/274660/images/8176_n.jpg",biography:"I am DrDamodar Paudel,currently working as consultant Physician in Nepal police Hospital.",institutionString:null,institution:null},{id:"241562",title:"Dr.",name:"Melvin",middleName:null,surname:"Sanicas",slug:"melvin-sanicas",fullName:"Melvin Sanicas",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/241562/images/6699_n.jpg",biography:null,institutionString:null,institution:null},{id:"337446",title:"Dr.",name:"Maria",middleName:null,surname:"Zavala-Colon",slug:"maria-zavala-colon",fullName:"Maria Zavala-Colon",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Puerto Rico, Medical Sciences Campus",country:{name:"United States of America"}}},{id:"338856",title:"Mrs.",name:"Nur Alvira",middleName:null,surname:"Pascawati",slug:"nur-alvira-pascawati",fullName:"Nur Alvira Pascawati",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Universitas Respati Yogyakarta",country:{name:"Indonesia"}}},{id:"441116",title:"Dr.",name:"Jovanka M.",middleName:null,surname:"Voyich",slug:"jovanka-m.-voyich",fullName:"Jovanka M. Voyich",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"Montana State University",country:{name:"United States of America"}}},{id:"330412",title:"Dr.",name:"Muhammad",middleName:null,surname:"Farhab",slug:"muhammad-farhab",fullName:"Muhammad Farhab",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Agriculture Faisalabad",country:{name:"Pakistan"}}},{id:"349495",title:"Dr.",name:"Muhammad",middleName:null,surname:"Ijaz",slug:"muhammad-ijaz",fullName:"Muhammad Ijaz",position:null,profilePictureURL:"//cdnintech.com/web/frontend/www/assets/author.svg",biography:null,institutionString:null,institution:{name:"University of Veterinary and Animal Sciences",country:{name:"Pakistan"}}}]}},subseries:{item:{id:"86",type:"subseries",title:"Business and Management",keywords:"Demographic shifts, Innovation, Technology, Next-gen leaders, Worldwide environmental issues and clean technology, Uncertainty and political risks, Radical adjacency, Emergence of new business ecosystem type, Emergence of different leader and leader values types, Universal connector, Elastic enterprise, Business platform, Supply chain complexity",scope:"